Database: MEDLINE <: biomedical, nursing & dental literature, 1966 - Nov 2000.> Search Strategy (You Saved Citations 1-300 From Set 61): ----------------------------------------------------------------------------- 1 exp Tooth demineralization/ 22684 2 demineralization.mp. 1629 3 caries.mp. 15334 4 caires.mp. 1 5 craies.mp. 0 6 careis.mp. 4 7 carise.mp. 0 8 (teeth adj3 cavit:).mp. 422 9 (tooth adj3 cavit:).mp. 217 10 (dental adj3 cavit:).mp. 276 11 (dentin adj3 cavit:).mp. 256 12 (enamel adj3 cavit:).mp. 183 13 (teeth adj3 decay:).mp. 379 14 (tooth adj3 decay:).mp. 325 15 (dental adj3 decay:).mp. 251 16 (dentin adj3 decay:).mp. 12 17 (enamel adj3 decay:).mp. 20 18 (active adj decay).mp. 9 19 (rampant adj3 decay:).mp. 14 20 (recurrent adj3 decay:).mp. 30 21 (white adj spot:).mp. 513 22 carious.mp. 2083 23 cariology.ti,ab. 56 24 (non-cavitated adj3 lesion:).mp. 15 25 (noncavitated adj3 lesion:).mp. 2 26 Tooth remineralization/ 479 27 (dental adj3 fissure:).mp. 99 28 (tooth adj3 fissure:).mp. 50 29 (teeth adj3 fissure:).mp. 98 30 caries-free.mp. 606 31 cariesfree.mp. 17 32 Cariogenic agents/ 729 33 precavit:.mp. 8 34 (filled adj3 teeth).mp. 513 35 (filled adj3 tooth).mp. 117 36 (oral adj fissure:).mp. 6 37 (tooth adj3 remineraliz:).mp. 28 38 (teeth adj3 remineraliz:).mp. 24 39 dft.mp. 415 40 dfs.mp. 1266 41 dmf:.mp. 6412 42 cariogeni:.mp. 1789 43 or/1-42 32343 44 exp Genetics/ 1206180 45 (cn or ge).fs. 901090 46 gene$1.mp. 443661 47 genetic:.mp. 259943 48 genom:.mp. 99495 49 genotyp:.mp. 48882 50 chromosom:.mp. 199850 51 congenit:.mp. 84285 52 familial.mp. 37802 53 heritab:.mp. 5915 54 inherit:.mp. 33803 55 twin$1.mp. 19323 56 Diseases in twins/ 8030 57 exp Multiple birth offspring/ 12476 58 consanguin:.mp. 6420 59 or/44-58 1589028 60 43 and 59 1434 61 limit 60 to english language 1251 62 from 61 keep 1-300 300 *************************** <1> UI - 20341505 AU - Acil Y AU - Terheyden H AU - Dunsche A AU - Fleiner B AU - Jepsen S IN - Department for Oral and Maxillofacial Surgery, Kiel University, Arnold-Heller-Strasse 16, 24105 Kiel, Germany. acil@mkg.uni-kiel.de TI - Three-dimensional cultivation of human osteoblast-like cells on highly porous natural bone mineral. SO - Journal of Biomedical Materials Research 2000 Sep 15;51(4):703-10 AB - In this study, we investigated the growth and extracellular matrix synthesis of human osteoblast-like cells on highly porous natural bone mineral. Human bone cells were isolated from trabecular bone during routine iliac crest biopsies. Under conventional culture conditions, trabecular bone cells were able to assume the organization of a three-dimensional structure on a porous natural bone mineral (Bio-Oss(R) Block). Scanning electron microscopy examination after 6 weeks revealed multiple cell layers on the trabecular block. Transmission electron microscopy examination after 6 weeks revealed the accumulation of mature collagen fibrils in the intracellular and extracellular spaces, and showed multilayered, rough endoplasmic reticulum as well as mitochondria-rich cells surrounded by dense extracellular matrix. These morphological observations suggest that the cell layer may resemble the natural three-dimensional structure. Biochemical analysis revealed that the hydroxylysylpyridinoline, lysylpyridinoline, and hydroxyproline content of the cell layer increased in a time-dependent manner, whereas in monolayer culture without natural bone mineral, no measurable amounts of hydroxylysylpyridinoline or lysylpyridinoline, and a barely measurable amount of hydroxyproline, were noted. Mature collagen extracted by ethylenediaminetetraacetic acid-demineralization from the cell layer on natural bone mineral showed an identical electrophoretic pattern to that observed in human bone, as evaluated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The present study demonstrated an excellent biocompatibility of the highly porous natural bone mineral in a three-dimensional bone cell culture system, and thus its potential for tissue-engineered growth of human bone. <2> UI - 20340960 AU - Honeyman AL AU - Curtiss R 3rd IN - Department of Biology, Washington University, St Louis, MO 63130, USA. TI - The mannitol-specific enzyme II (mtlA) gene and the mtlR gene of the PTS of Streptococcus mutans. SO - Microbiology 2000 Jul;146 ( Pt 7):1565-72 AB - The phosphoenolpyruvate-dependent phosphotransferase system (PTS) is widely found among Gram-positive bacteria. It is the major source of carbohydrate transport in the dental pathogen Streptococcus mutans. The transported carbohydrates are fermented to produce large amounts of lactic acid which initiates dental caries. The authors have isolated the S. mutans gene for the mannitol-specific Enzyme II (EII) component of the PTS, mtlA, and the adjacent mtlR gene, which is located in the same operon. The mtlR gene is located between mtlA and the genes mtlF and mtlD. The nucleotide sequence of the mtlA and mtlR loci has been determined. The deduced mtlA gene product of S. mutans consists of 589 amino acids with a molecular mass of 62.0 kDa. It exhibits similarity with the mtlA gene products from other organisms. However, the similarity between these proteins is generally restricted to the 470 amino-terminal residues of the S. mutans protein. This region would correspond to the EIICB domains of the PTS. The authors have previously shown that the S. mutans mtlF gene product exhibits 76.6% similarity to the carboxyl-terminal 143 amino acids of the Escherichia coli mtlA product and that the mtlF gene encodes the EIIA domain of the PTS. Thus, the genes that encode the EIICB and the EIIA domains are separated by approximately 2250 bp. In many organisms, all of the EII domains may be fused together to form one molecule. The fact that these domains are separated by this distance in S. mutans supports the hypothesis that various functional domains of the PTS have been rearranged during evolution. The sequence of the 119 carboxyl-terminal amino acids of the S. mutans mtlA gene product also displays homology to the carboxyl-terminal end of the EIIB domain of various mannitol PTSs. Thus, this domain may have been duplicated in S. mutans during evolution of the operon. The mtlR gene is located in the same operon structure as mtlA but these loci are separated by an intragenic space. The precise 5' end of the mtlR locus cannot be determined either by in vitro transcription-translation assays or based upon nucleotide sequence analysis because of the apparent lack of a ribosome-binding site preceding the gene. The deduced mtlR gene product, which consists of approximately 650 amino acids with a molecular mass of 75.3 kDa, exhibits limited similarity to several potential transcriptional regulators. However, the exact function of this locus is currently unknown. <3> UI - 20359334 AU - Kitten T AU - Munro CL AU - Michalek SM AU - Macrina FL IN - Philips Institute of Oral & Craniofacial Molecular Biology, Virginia Commonwealth University, Richmond, Virginia 23298, USA. tkitten@hsc.vcu.edu TI - Genetic characterization of a Streptococcus mutans LraI family operon and role in virulence. SO - Infection & Immunity 2000 Aug;68(8):4441-51 AB - Proteins belonging to the LraI (for "lipoprotein receptor antigen") family function as adhesins in several streptococci, as a virulence factor for endocarditis in at least one of these species, and potentially as metal transporters in many bacteria. We have identified and characterized the chromosomal locus containing the LraI family gene (designated sloC) from Streptococcus mutans, an agent of dental caries and endocarditis in humans. Northern blot analysis indicated that sloC is cotranscribed with three other genes. As with other LraI operons, the sloA and sloB genes apparently encode components of an ATP-binding cassette transport system. The product of the fourth gene, sloR, has homology to the metal-dependent regulator from Corynebacterium diphtheriae, DtxR. A potential binding site for SloR was identified upstream from the sloABCR operon and was conserved upstream from LraI operons in several other streptococci. Potential SloR homologs were identified in the unfinished genomic sequences from two of these, S. pneumoniae and S. pyogenes. Mutagenesis of sloC in S. mutans resulted in apparent loss of expression of the entire operon as assessed by Northern blot analysis. The sloC mutant was indistinguishable from its wild-type parent in a gnotobiotic rat model of caries but was significantly less virulent in a rat model of endocarditis. Virulence for endocarditis was restored by correction of the sloC mutation but not by provision of the sloC gene in trans, suggesting that virulence requires the expression of other genes in the sloC operon. <4> UI - 20357036 AU - Aydin ZD AU - Barista I AU - Canpinar H AU - Sungur A AU - Tekuzman G IN - Department of Internal Medicine, Hacettepe University Faculty of Medicine, Sihhiye, Ankara, Turkey. TI - Gastric lymphomas in Turkey. Analysis of prognostic factors with special emphasis on flow cytometric DNA content. SO - Cancer 2000 Jul 1;89(1):12-20 AB - BACKGROUND: In contrast to DNA ploidy, to the authors' knowledge the prognostic significance of S-phase fraction (SPF) in gastric lymphomas has not been determined. In the current study, the prognostic significance of various parameters including SPF and DNA aneuploidy were analyzed and some distinct epidemiologic and biologic features of gastric lymphomas in Turkey were found. METHODS: A series of 78 gastric lymphoma patients followed at Hacettepe University is reported. DNA flow cytometry was performed for 34 patients. The influence of various parameters on survival was investigated with the log rank test. The Cox proportional hazards model was fitted to identify independent prognostic factors. RESULTS: The median age of the patients was 50 years. There was no correlation between patient age and tumor grade. DNA content analysis revealed 4 of the 34 cases to be aneuploid with DNA index values < 1.0. The mean SPF was 33.5%. In the univariate analysis, surgical resection of the tumor, modified Ann Arbor stage, performance status, response to first-line chemotherapy, lactate dehydrogenase (LDH) level, and SPF were important prognostic factors for disease free survival (DFS). The same parameters, excluding LDH level, were important for determining overall survival (OS). In the multivariate analysis, surgical resection of the tumor, disease stage, performance status, and age were found to be important prognostic factors for OS. CONCLUSIONS: To the authors' knowledge the current study is the first to demonstrate the prognostic significance of SPF in gastric lymphomas. The distinguishing features of Turkish gastric lymphoma patients are 1) DNA indices of aneuploid cases that all are < 1.0, which is a unique feature; 2) a lower percentage of aneuploid cases; 3) a higher SPF; 4) a younger age distribution; and 5) lack of an age-grade correlation. The authors conclude that gastric lymphomas in Turkey have distinct biologic and epidemiologic characteristics. Copyright 2000 American Cancer Society. <5> UI - 20399327 AU - Yoder S AU - Cao C AU - Ugen KE AU - Dao ML IN - Department of Biology, University of South Florida, Tampa, USA. TI - High-level expression of a truncated wall-associated protein A from the dental cariogenic Streptococcus mutans. SO - DNA & Cell Biology 2000 Jul;19(7):401-8 AB - Streptococcus mutans plays a primary role in the formation of dental caries. Previously, in our laboratory, an S. mutans genomic library was prepared, and the wapA gene was cloned into the shuttle vector, pSA4/4B2. To generate overexpression of wapA and to facilitate efficient purification of the WapA protein for use as an immunogen, an expression vector with the strong tac promoter was used. In order to answer questions regarding the optimization of solubility and expression based on gene size or the hydrophobicity of the protein product, 12 truncated constructs of the wapA gene were prepared using PCR. The truncated products were subcloned into the pGEX-6P-1 glutathione S-transferase (GST) fusion vector and expressed in E. coli BL21. The fusion proteins were analyzed by SDS-PAGE and confirmed by analysis with anti-GST and anti-WapA antibodies. Our study suggests that abrogation of the wapA promoter is necessary for expression of this gene in this expression system. Deletion of the signal peptide and the hydrophobic C terminus of WapA increased expression compared with the full-length construct, and truncation at the protease cleavage site of the C-terminal region greatly increased the stability of the protein without a loss in reactivity with the anti-WapA antibody. Western immunoblot analysis with anti-WapA antiserum clearly showed that the majority of the epitopes of the GST-WapA fusions are located in the N-terminal region of WapA. The immunogenicity of the various WapA fusion products is being examined in mice and rats to further map the immunologically dominant regions of the protein. This method effectively increased the expression of WapA and should contribute to the further understanding of gene expression of E. coli, as well as aid in the characterization of this protein for future immunologic evaluation. <6> UI - 20373071 AU - Simmonds RS AU - Tompkins GR AU - George RJ IN - Department of Microbiology, University of Otago, Dunedin. TI - Dental caries and the microbial ecology of dental plaque: a review of recent advances. [Review] [68 refs] SO - New Zealand Dental Journal 2000 Jun;96(424):44-9 AB - Our understanding of the microbial ecology of dental plaque has rapidly grown with recent developments in the techniques of molecular biology. In particular, knowledge of the mechanisms underlying the acquisition, establishment, pathogenicity, and evolution of the group of organisms responsible for dental caries--the mutans streptococci--has expanded to the point that we can now contemplate new opportunities for caries prevention. These advances reinforce developing concepts of dental plaque as an interdependent, interacting community of specialised organisms with an ability to rapidly adapt conferred by gene structures that facilitate the expeditious modular rearrangement of protein components. [References: 68] <7> UI - 20314570 AU - Ochs RL AU - Muro Y AU - Si Y AU - Ge H AU - Chan EK AU - Tan EM IN - W. M. Keck Autoimmune Disease Center, Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA 92037, USA. TI - Autoantibodies to DFS 70 kd/transcription coactivator p75 in atopic dermatitis and other conditions. SO - Journal of Allergy & Clinical Immunology 2000 Jun;105(6 Pt 1):1211-20 AB - BACKGROUND: Sera of patients with atopic dermatitis (AD) were found to have autoantibodies that reacted with tissue culture cell substrates in immunohistochemistry to display a characteristic pattern of nuclear distribution of dense fine speckles. The sera also recognized a 70-kd protein on Western immunoblots, and the antigen was termed dense fine speckles 70 kd (DSF70). OBJECTIVE: Because spontaneously occurring autoantibodies could be immune responses to proteins that might be participating in the disease process, it was of interest to identify the antigens driving the autoimmune antibody response. METHODS: A serum containing high-titer antibodies to DFS70 was used to immunoscreen a complementary (c)DNA expression library to isolate cDNA encoding the antigen. After the cDNA was isolated, this was used to express recombinant protein to determine the prevalence of antibody in AD and other conditions. RESULTS: Thirty percent of patients with AD were found to have antibody to recombinant DFS70 in Western immunoblots. Sixteen percent of patients with asthma and 9% of patients with interstitial cystitis had antibodies of the same specificities. The cDNA encoding DFS70 was identical to a transcription coactivator called p75, which had been shown to be required for RNA polymerase II-dependent transcription. Another important finding was that IgE antibodies to DFS70 were also present in AD sera. CONCLUSION: It is suggested that a common basis for the presence of autoantibodies to DFS70 might be related to AD in asthma, interstitial cystitis, and other conditions. A possible role of this antigen-antibody system in pathogenesis remains to be demonstrated, but it appears to be a marker for a subset of patients with AD. <8> UI - 20275053 AU - Gregory RK AU - Powles TJ AU - Salter J AU - Chang JC AU - Ashley S AU - Dowsett M IN - Breast Unit, Royal Marsden Hospital, Sutton, Surrey, UK. TI - Prognostic relevance of cerbB2 expression following neoadjuvant chemotherapy in patients in a randomised trial of neoadjuvant versus adjuvant chemoendocrine therapy. SO - Breast Cancer Research & Treatment 2000 Jan;59(2):171-5 AB - Recent advances in the detection and treatment of breast cancer have led to an intensive search for new markers of both prognosis and chemoresponsiveness. The oncogene cerbB2 has proved to be one of the most promising markers currently under study, both as a predictor of chemoresponsiveness and as a marker of poor prognosis. In addition the increasing use of neoadjuvant chemotherapy has led to the loss of standard prognostic criteria. In order to study the potential role of cerbB2 expression as an indicator of chemoendocrine resistance and poor prognosis, both before and after chemotherapy, we obtained tumour sections from 283 women enrolled onto a neoadjuvant trial. In this trial patients were randomised to receive either primary surgery followed by adjuvant chemoendocrine treatment or neoadjuvant chemoendocrine therapy followed by surgery. CerbB2 status was determined immunohistochemically on all of these patients. Thirty-eight percent of the tumours were cerbB2 positive. There was no significant difference in expression between the adjuvant (41%) and neoadjuvant arms (35%). CerbB2 positive patients were much more likely to have shown non-response to chemoendocrine therapy (p < 0.001) and had a worse DES (p < 0.05). The best prognosis was seen in cerbB2 negative patients receiving neoadjuvant chemoendocrine therapy who showed a significantly better DFS (p < 0.05), than the cerbB2 negative patients receiving adjuvant therapy. <9> UI - 20275047 AU - Goodson WH 3rd AU - Moore DH 2nd AU - Ljung BM AU - Chew K AU - Mayall B AU - Smith HS AU - Waldman FM IN - Department of Surgery, California Pacific Medical Research Institute, San Francisco, USA. TI - The prognostic value of proliferation indices: a study with in vivo bromodeoxyuridine and Ki-67. SO - Breast Cancer Research & Treatment 2000 Jan;59(2):113-23 AB - Proliferation indices are intended to help patients and clinicians make treatment decisions. We have previously demonstrated that a proliferation index based on in vivo labeling of S-phase cells with bromodeoxyuridine (BrdUrd) correlates with Ki-67 labeling index (LI). We now compare the prognostic value of these indices. With written consent, we gave 129 women with biopsy confirmed breast cancer 200 mg/M2 BrdUrd during 30 min immediately preceding surgery. We used IU-4 anti BrdUrd antibody to count the immunohistochemical labeling index (LI) of DNA-incorporated BrdUrd in 2,000 cells and MIB-1 to count Ki-67 (118 cases). Patients received standard surgical and adjuvant treatment. No patients were lost to follow-up and patients were followed a minimum of 2 (median 5.1) years. We compared survival and recurrence in tumors with high vs low labeling indices. We found that women in the low BrdUrd LI group had better disease free survival (92% vs 67% 5-yr DFS p = 0.001) and overall survival (94% vs 70% 5-yr OS, p = 0.0001) than those with a high LI. In comparison, a low Ki-67 index predicted better OS (87% vs 80% 5-yr OS, p = 0.020) and a trend for better DFS (84% vs 72% DFS p = 0.055). The apparent superiority of BrdUrd LI over Ki-67 LI is likely due to chance (p = 0.18). In multivariate survival analyses we found that BrdUrd LI proliferative index significantly improves prediction of DFS or OS even when node status, age or tumor size is in the model. We conclude that markers of proliferation are useful adjuncts in predicting patient prognosis. <10> UI - 20265403 AU - Schwartz Z AU - Lohmann CH AU - Wieland M AU - Cochran DL AU - Dean DD AU - Textor M AU - Bonewald LF AU - Boyan BD IN - Department of Orthopedics, University of Texas Health Science Center, San Antonio 78229-3900, USA. TI - Osteoblast proliferation and differentiation on dentin slices are modulated by pretreatment of the surface with tetracycline or osteoclasts. SO - Journal of Periodontology 2000 Apr;71(4):586-97 AB - BACKGROUND: Implant surface roughness and chemical composition, as well as other factors, affect the ability of osteogenic cells to form bone adjacent to an implant. The same principles may also apply to the tooth root and some reports have shown that surface modification of the root may lead to improved restoration of the periodontal apparatus. The most common of these surface modification techniques involves demineralization with citric acid or treatment with tetracycline to expose collagen fibrils. In addition, during normal bone remodeling, osteoclasts demineralize the extracellular matrix, leaving resorption pits and exposed collagen fibrils. In this study, the effect of different dentin surface-preparation techniques on osteoblasts were compared. METHODS: Slices of sperm whale dentin were mechanically polished and surfaces were treated with tetracycline-HCl (TCN) or were cultured with mouse bone marrow cells to create a surface with osteoclast (OC) resorption pits or left untreated. Profilometry, x-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM) were used to evaluate the 3 different dentin surfaces. MG63 osteoblast-like cells were cultured on the 3 different surfaces and the effect of dentin surface preparation technique on MG63 cell proliferation (cell number), differentiaton (alkaline phosphatase specific activity of isolated cells and cell layer lysates; osteocalcin production), and local factor production (transforming growth factor (TGF)-beta1 and prostaglandin E2 (PGE2) compared. RESULTS: Profilometry showed the polished and TCN surfaces were smooth with comparable Ra values, whereas the OC surfaces were slightly rougher due to resorption pits which covered 3.7% of the surface. XPS measurements showed that TCN treatment reduced the Ca and P content of the surface, indicating that it had dissolved the mineral. Osteoclast-resorption also reduced the Ca and P content, but to a lesser extent. MG63 cell proliferation on polished dentin and tissue culture polystyrene was equivalent. In contrast, cells grown on the TCN- and OC-treated surfaces exhibited increased proliferation. No effect of surface treatment on cell alkaline phosphatase activity was observed, but activity in the cell layer lysates was increased on the TCN- and OC-treated surfaces. Osteocalcin production was reduced on all dentin surfaces, but the greatest reduction was found on the TCN-treated surface. Production of both TGF-beta1 and PGE2 was increased on the treated surfaces. All effects were greatest in cultures grown on the TCN-treated dentin. CONCLUSIONS: These data indicate that demineralization of the dentin surface promotes proliferation of osteoblasts and early differentiation events like production of alkaline phosphatase and autocrine mediators such as PGE2 and TGF-beta1. However, later differentiation events like osteocalcin production are decreased. Osteoclast-mediated bone resorption elicits similar responses; less than 4% of the dentin surface resulted in approximately 75% of the response caused by TCN treatment. These observations suggest that greater attention should be paid to the effects of osteoclastic resorption in designing methods for enhancing bone and cementum formation adjacent to root surfaces. <11> UI - 20269630 AU - Trope C AU - Kaern J AU - Hogberg T AU - Abeler V AU - Hagen B AU - Kristensen G AU - Onsrud M AU - Pettersen E AU - Rosenberg P AU - Sandvei R AU - Sundfor K AU - Vergote I IN - Department of Gynecologic Oncology, The Norwegian Radium Hospital, Oslo. claes.trope@klinmed.uio.no TI - Randomized study on adjuvant chemotherapy in stage I high-risk ovarian cancer with evaluation of DNA-ploidy as prognostic instrument [see comments]. CM - Comment in: Ann Oncol 2000 Mar;11(3):259-61 SO - Annals of Oncology 2000 Mar;11(3):281-8 AB - PURPOSE: Adjuvant chemotherapy versus observation and chemotherapy at progression was evaluated in 162 patients in a prospective randomized multicenter study. We also evaluated DNA-measurements as an additional prognostic factor. PATIENTS AND METHODS: Patients received adjuvant carboplatin AUC 7 every 28 days for six courses (n = 81) or no adjuvant treatment (n = 81). Eligibility included surgically staged and treated patients with FIGO stage I disease, grade 1 aneuploid or grade 2 or 3 non-clear cell carcinomas or clear cell carcinomas. Disease-free (DFS) and disease-specific (DSS) survival were end-points. RESULTS: Median follow-up time was 46 months and progression was observed in 20 patients in the treatment group and 19 in the control group. Estimated five-year DFS and DSS were 70% and 86% in the treatment group and 71% and 85% in the control group. The hazard ratio was 0.98 (95% confidence interval (95% CI): 0.52-1.83) regarding DFS and 0.94 (95% CI: 0.37-2.36) regarding DSS. No significant differences in DFS or DSS could be seen when the log-rank test was stratified for prognostic variables. Therefore, data from both groups were pooled for the analysis of prognostic factors. DNA-ploidy (P = 0.003), extracapsular growth (P = 0.005), tumor rupture (P = 0.04), and WHO histologic grade (P = 0.04) were significant independent prognostic factors for DFS with P < 0.0001 for the model in the multivariate Cox analysis. FIGO substage (P = 0.01), DNA ploidy (P < 0.05), and histologic grade (P = 0.05) were prognostic for DSS with a P-value for the model < 0.0001. CONCLUSIONS: Due to the small number of patients the study was inconclusive as regards the question of adjuvant chemotherapy. The survival curves were superimposable, but with wide confidence intervals. DNA-ploidy adds objective independent prognostic information regarding both DFS and DSS in early ovarian cancer. <12> UI - 99326635 AU - Schafer TE AU - Lapp CA AU - Hanes CM AU - Lewis JB AU - Wataha JC AU - Schuster GS IN - Department of Pediatric Dentistry, Medical College of Georgia, Augusta, Georgia 30912, USA. TI - Estrogenicity of bisphenol A and bisphenol A dimethacrylate in vitro. SO - Journal of Biomedical Materials Research 1999 Jun 5;45(3):192-7 AB - Although pit and fissure sealants have been utilized extensively in dentistry as a way of preventing occlusal caries, results described by Olea et al. (1996) raised concerns about the safety of sealants and other resin-based dental materials due to the reported presence of bisphenol A (BPA) and its dimethacrylate ester (BPA-DM). Although the release of these compounds from dental materials has not been substantiated by two subsequent studies, we believed it was important to confirm or refute the report that BPA and BPA-DM have estrogenic activity in vitro. We grew breast cancer cells (MCF-7, T-47D, ZR-75-1) known to proliferate under estrogenic stimulation in phenol red-free DMEM containing human serum and concentrations of BPA or BPA-DM ranging from 10(-8)M to 5 x 10(-6)M. After 1 week, plates were harvested for crystal violet or sulforhodamine-B assays, and the optical densities of groups of treated cells were compared with values from control cells. At concentrations at or above 10(-6)M, both BPA and BPA-DM significantly increased cell proliferation (p < 0.05), comparable to the increase seen with 10(-9)M of estrogen. Flow cytometric methods demonstrated that these mitogenic effects occurred within 24 h of exposure to estrogen, BPA, or BPA-DM. The increase in DNA synthesis was analogous to that seen with estrogen stimulation. Thus, we confirmed that BPA and BPA-DM cause cell proliferation at micromolar concentrations that exceed the effective concentrations of estrogen by 1 to 10,000-fold. Copyright 1999 John Wiley & Sons, Inc. <13> UI - 20316028 AU - Caufield PW AU - Dasanayake AP AU - Li Y AU - Pan Y AU - Hsu J AU - Hardin JM IN - School of Dentistry, University of Alabama, Birmingham, Alabama 35294, USA. page@uab.edu TI - Natural history of Streptococcus sanguinis in the oral cavity of infants: evidence for a discrete window of infectivity. SO - Infection & Immunity 2000 Jul;68(7):4018-23 AB - The heterogeneous group of oral bacteria within the sanguinis (sanguis) streptococci comprise members of the indigenous biota of the human oral cavity. While the association of Streptococcus sanguinis with bacterial endocarditis is well described in the literature, S. sanguinis is thought to play a benign, if not a beneficial, role in the oral cavity. Little is known, however, about the natural history of S. sanguinis and its specific relationship with other oral bacteria. As part of a longitudinal study concerning the transmission and acquisition of oral bacteria within mother-infant pairs, we examined the initial acquisition of S. sanguinis and described its colonization relative to tooth emergence and its proportions in plaque and saliva as a function of other biological events, including subsequent colonization with mutans streptococci. A second cohort of infants was recruited to define the taxonomic affiliation of S. sanguinis. We found that the colonization of the S. sanguinis occurs during a discrete "window of infectivity" at a median age of 9 months in the infants. Its colonization is tooth dependent and correlated to the time of tooth emergence; its proportions in saliva increase as new teeth emerge. In addition, early colonization of S. sanguinis and its elevated levels in the oral cavity were correlated to a significant delay in the colonization of mutans streptococci. Underpinning this apparent antagonism between S. sanguinis and mutans streptococci is the observation that after mutans streptococci colonize the infant, the levels of S. sanguinis decrease. Children who do not harbor detectable levels of mutans streptococci have significantly higher levels of S. sanguinis in their saliva than do children colonized with mutans streptococci. Collectively, these findings suggest that the colonization of S. sanguinis may influence the subsequent colonization of mutans streptococci, and this in turn may suggest several ecological approaches toward controlling dental caries. <14> UI - 20233833 AU - Ohta H AU - Yomogida K AU - Dohmae K AU - Nishimune Y IN - Department of Science for Laboratory Animal Experimentation, Research Institute for Microbial Diseases, Osaka University, Yamadaoka 3-1, Suita, Osaka, Japan. TI - Regulation of proliferation and differentiation in spermatogonial stem cells: the role of c-kit and its ligand SCF. SO - Development 2000 May;127(10):2125-31 AB - To study self-renewal and differentiation of spermatogonial stem cells, we have transplanted undifferentiated testicular germ cells of the GFP transgenic mice into seminiferous tubules of mutant mice with male sterility, such as those dysfunctioned at Steel (Sl) locus encoding the c-kit ligand or Dominant white spotting (W) locus encoding the receptor c-kit. In the seminiferous tubules of Sl/Sl(d) or Sl(17H)/Sl(17H) mice, transplanted donor germ cells proliferated and formed colonies of undifferentiated c-kit (-) spermatogonia, but were unable to differentiate further. However, these undifferentiated but proliferating spermatogonia, retransplanted into Sl (+) seminiferous tubules of W mutant, resumed differentiation, indicating that the transplanted donor germ cells contained spermatogonial stem cells and that stimulation of c-kit receptor by its ligand was necessary for maintenance of differentiated type A spermatogonia but not for proliferation of undifferentiated type A spermatogonia. Furthermore, we have demonstrated that their transplantation efficiency in the seminiferous tubules of Sl(17H)/Sl(17H) mice depended upon the stem cell niche on the basement membrane of the recipient seminiferous tubules and was increased by elimination of the endogenous spermatogonia of mutant mice from the niche by treating them with busulfan. <15> UI - 20293353 AU - Sperandio M AU - Weber L AU - Jauch A AU - Janssen B AU - Mehls O AU - Schaefer F IN - Department of Pediatrics and. Institute of Human Genetics, University of Heidelberg, Germany. TI - Cutaneous white spots in a child with polycystic kidneys: a clue to TSC2/PKD1 gene mutation. SO - Nephrology, Dialysis, Transplantation 2000 Jun;15(6):909-12 <16> UI - 20322334 AU - Thomas X AU - Dombret H AU - Cordonnier C AU - Pigneux A AU - Gardin C AU - Guerci A AU - Vekhoff A AU - Sadoun A AU - Stamatoullas A AU - Fegueux N AU - Maloisel F AU - Cahn JY AU - Reman O AU - Gratecos N AU - Berthou C AU - Huguet F AU - Kotoucek P AU - Travade P AU - Buzyn A AU - de Revel T AU - Vilque JP AU - Naccache P AU - Chomienne C AU - Degos L AU - Fenaux P IN - Department of Hematology, Hopital Edouard Herriot, Lyon, France. TI - Treatment of relapsing acute promyelocytic leukemia by all-trans retinoic acid therapy followed by timed sequential chemotherapy and stem cell transplantation. APL Study Group. Acute promyelocytic leukemia. SO - Leukemia 2000 Jun;14(6):1006-13 AB - The purpose of this study was to assess the safety and efficacy of stem cell transplantation (SCT) mainly autologous SCT as consolidation therapy in APL patients who relapsed and achieved a second complete remission (CR2). Fifty adult patients with a first relapsed APL, of whom 39 had been previously treated with ATRA, entered a multicenter trial of oral ATRA until complete remission (CR) achievement followed by timed sequential chemotherapy (EMA combining etoposide 200 mg/m2/day for 3 days, mitoxantrone 12 mg/m2/day for 3 days, and cytarabine 500 mg/m2/day for two sequences of 3 days). EMA was started either after CR achievement, or on day 1 of ATRA because of initial white blood cell (WBC) counts >5 x 10(9)/l, or rapidly added to ATRA in order to prevent ATRA syndrome because WBC count increased under ATRA. Forty-five patients (90%, 95% CI 78%-97%) were in CR after induction therapy. Five patients died from infection during aplasia following EMA chemotherapy. Eleven patients who achieved CR had a familial HLA-identical donor and were allografted. The median disease-free survival (DFS) of allografted patients was 8.2 months. The 34 other CR patients were scheduled for autologous peripheral blood (PB) SCT (intent-to-treat group). Actually, autologous transplantation was only carried out in 22 patients (65%) (17 PBSCT and five autologous bone marrow transplantation (BMT)). Reasons for not autografting were early relapse (three patients), severe toxicity of EMA chemotherapy (six patients), and refusal or failure of stem cell harvest (three patients). The 3-year DFS rate of patients actually autografted was 77%. Among the 17 autografted patients still in CR2, nine patients have already reached a longer CR2 than first CR (CR1). Results of detection of PML/RARalpha by RT-PCR after autologous transplantation show negative findings in eight of the nine patients tested. We conclude that (1) ATRA combined to EMA chemotherapy is effective in the treatment of relapsed APL; (2) allogeneic BMT may be too toxic after salvage treatment including EMA intensive chemotherapy; (3) clinical outcome of autografted patients and preliminary molecular results regarding detection of PML/RARalpha after autologous PBSCT are encouraging. <17> UI - 20237498 AU - Li Y AU - Wang W AU - Caufield PW IN - Department of Oral Biology, School of Dentistry, University of Alabama at Birmingham, Ala., USA. yihongli@uab.edu TI - The fidelity of mutans streptococci transmission and caries status correlate with breast-feeding experience among Chinese families. SO - Caries Research 2000 Mar-Apr;34(2):123-32 AB - Our previous study reported that the fidelity of mutans streptococci (MS) transmission from mother to infant was gender- and race-specific within a Birmingham, Ala., population. We hypothesized that fidelity might be a function of postnatal nurturing practices. The purpose of the present study was to investigate those factors that could be correlated with MS transmission among Chinese children whose nurturing histories were known. Forty-eight families with 2- to 3-year-old children were selected from two kindergartens of Beijing, China. A questionnaire concerning the childhood nursing practice was obtained from the parents. Dental caries status was examined, and bacterial samples were collected for all participants twice at 6-month intervals. An average of six isolates of MS was picked at random from selective medium from each individual at each visit. Chromosomal DNA fingerprints were performed for all MS isolates to determine the genomic similarity within each family and among individuals. The results showed that 66% of the children at 2-3 years of age harbored MS, and 46% were caries-active. Of those children, 70% were breast-fed. Among the children whose chromosomal DNA fingerprint genotypes of MS matched that of their mothers, 88% were breast-fed compared with only 12% who were not breast-fed (p = 0.03). We also found that children who were breast-fed for more than 9 months were likely to harbor strains of MS common to their mothers (p = 0.04) and experience more dental caries (dmft = 4.4) at 3 years of age compared with children who were breast-fed less than 9 months (dmft = 1.4, p = 0.04). The data suggest that breast-feeding, especially prolonged breast-feeding, may correlate with the fidelity of transmission and that prolonged breast-feeding may contribute to a higher caries rate. <18> UI - 20270569 AU - Ball ED AU - Wilson J AU - Phelps V AU - Neudorf S IN - Department of Medicine, University of California San Diego, La Jolla, CA 92093-0960, USA. TI - Autologous bone marrow transplantation for acute myeloid leukemia in remission or first relapse using monoclonal antibody-purged marrow: results of phase II studies with long-term follow-up. SO - Bone Marrow Transplantation 2000 Apr;25(8):823-9 AB - One hundred and thirty-eight patients with AML underwent ABMT with monoclonal antibody plus complement-purged marrow between August 1984 and March 1997. One hundred and ten patients were in CR (CR1: 23; CR2/3: 87) and 28 were in first relapse (R1) at ABMT. Preparative regimens included busulfan (16 mg/kg) and CY (120 mg/kg) (n = 93), CY (120 mg/kg over 2 days) with TBI (1200 cGy) (n = 35), and busulfan (16 mg/kg) plus etoposide (60 mg/kg) (n = 10). CR1 patients treated with CY/TBI (n = 7) had 3- and 5-year disease-free survival (DFS) rates of 71% and 57%. CR1 patients treated with BU/CY (n = 12), had 3- and 5-year DFS rates of 45%. Three and 5-year DFS for CR2/3 patients treated with CY/TBI (n = 26) was 23%. Three- and 5-year DFS for patients in CR2/3 treated with BU/CY (n = 55) was 31 and 28%. Three- and 5-year DFS for patients in R1 treated with BU/CY (n = 26) was 37%. In multivariate analysis, increased age was associated with greater risk of death and relapse. For CR2/3 patients, the length of CR1 was a significant predictor of DFS. ABMT performed in CR or R1 results in excellent 5-year DFS and OS. The contribution of purging may require a randomized trial comparing purged vs unpurged stem cell infusions. <19> UI - 20239717 AU - Midorikawa K AU - Murata M AU - Oikawa S AU - Tada-Oikawa S AU - Kawanishi S IN - Department of Hygiene, Mie University School of Medicine, Tsu, Mie 514-8507, Japan. TI - DNA damage by dimethylformamide: role of hydrogen peroxide generated during degradation. SO - Chemical Research in Toxicology 2000 Apr;13(4):309-15 AB - Dimethylformamide (DMF) has been suspected to associate with cancers in exposed workers, whereas there has been inadequate evidence for carcinogenicity in experimental animals. We demonstrated that H(2)O(2) was generated during the degradation of DMF under aerobic conditions, and that the amount of H(2)O(2) was enhanced by exposure to solar light or by the contamination of trace metal. Experiments using (32)P-5'-end-labeled DNA fragments revealed that the degraded DMF induced DNA damage in the presence of Cu(II). However, purified DMF did not induce DNA damage even in the presence of Cu(II). Addition of purified DMF enhanced DNA damage induced by H(2)O(2) in the presence of Cu(II). The degraded DMF caused Cu(II)-mediated DNA cleavage frequently at thymine and cytosine residues. The similar pattern of site-specific DNA damage was observed with purified DMF and H(2)O(2). Bathocuproine and catalase inhibited the DNA damage, indicating the involvement of Cu(I) and H(2)O(2). A typical free hydroxy radical scavenger showed no inhibitory effect on the DNA damage. Addition of purified DMF enhanced about 3-4-fold 8-oxo-7, 8-dihydro-2'-deoxyguanosine formation induced by H(2)O(2) and Cu(II). ESR spectroscopic study demonstrated that carbon-centered radicals and nitrogen-centered radicals were generated in the reaction mixture of DMF, H(2)O(2), and Cu(II). Inhibitory effects of scavengers on radical formation and DNA damage suggest that carbon-centered radicals and/or nitrogen-centered radicals may contribute to the DNA damage. These results suggest that H(2)O(2) generation during DMF degradation is related to the possible carcinogenic activity of DMF. <20> UI - 20206382 AU - Kirkham J AU - Robinson C AU - Strafford SM AU - Shore RC AU - Bonass WA AU - Brookes SJ AU - Wright JT IN - Division of Oral Biology, Leeds Dental Institute, Clarendon Way, Leeds, UK. orl.6jen@oralbio.novell.leeds.ac.uk TI - The chemical composition of tooth enamel in junctional epidermolysis bullosa. SO - Archives of Oral Biology 2000 May;45(5):377-86 AB - The junctionalis form of epidermolysis bullosa (EBJ) is associated with a number of clinical problems involving tooth enamel, including increased susceptibility to caries. The aim here was to carry out a chemical characterization of the enamel of teeth from EBJ patients compared with that of unaffected controls. The results showed that while protein concentration, amino acid composition and carbonate content were similar in both groups, EBJ enamel contained a significantly reduced mineral per volume content, resulting in enamel hypoplasia. In addition, Western blotting revealed the presence of serum albumin (a known inhibitor of enamel crystal growth) in EBJ enamel. This was not detected in control enamel or in enamel of teeth from patients with the dystrophic form of the disease. It is concluded that EBJ enamel is developmentally compromised and that the enamel defects are commensurate with the reported genetic lesions. <21> UI - 20289947 AU - Ayad M AU - Van Wuyckhuyse BC AU - Minaguchi K AU - Raubertas RF AU - Bedi GS AU - Billings RJ AU - Bowen WH AU - Tabak LA IN - Center for Oral Biology, Aab Institute of Biomedical Sciences, University of Rochester Medical Center, NY 14642, USA. TI - The association of basic proline-rich peptides from human parotid gland secretions with caries experience. SO - Journal of Dental Research 2000 Apr;79(4):976-82 AB - To address whether there are associations between the peptide composition of human parotid saliva and dental decay (caries) experience, we have characterized the peptides from parotid ductal saliva collected from nine adults who have remained free from dental caries (mean age = 59.2; Decayed Missing Filled Surfaces index [DMFS] = 0) and nine individuals who have experienced caries (mean age = 51.2; mean DMFS = 38.4). Ethanol-soluble peptides were size-fractionated on columns of Bio-Gel P-2; the salivary peptides derived from caries-susceptible subjects appeared larger than those found in the saliva of caries-free subjects. Peptides were then resolved into 19 species by cation exchange HPLC. Sequence analysis identified 18 peptides that appear to be proteolytic cleavage products of the basic proline-rich proteins IB-4, IB-5, IB-7, IB-8b, and P-B. The peptides that were more abundant in saliva obtained from the caries-free group differed from those isolated from the caries-susceptible group. The median peptide concentration of one possible precursor protein, IB-7, was found to be higher in saliva collected from caries-free individuals than in that from caries-susceptible individuals. Although differences were found in the phenotypes of proline-rich proteins expressed by these groups of caries-free and caries-susceptible subjects, no statistically significant associations were observed among proline-rich phenotypes and the level of any peptide. Collectively, our results indicate that proteolytic processing of parotid salivary proteins differs among individuals who have remained caries-free and those who have experienced dental decay. <22> UI - 20231807 AU - Taubman MA AU - Smith DJ AU - Holmberg CJ AU - Eastcott JW IN - Department of Immunology, The Forsyth Institute, Boston, Massachusetts 02115, USA. mtaubman@forsyth.org TI - Coimmunization with complementary glucosyltransferase peptides results in enhanced immunogenicity and protection against dental caries. SO - Infection & Immunity 2000 May;68(5):2698-703 AB - Peptide constructs from the catalytic (CAT) and glucan-binding (GLU) regions of the mutans streptococcal glucosyltransferase enzymes (GTF) can provide immunity to dental caries infection. A strategy of coimmunization was tested to determine whether protection could be enhanced. Rats were immunized with one of the previously described peptide constructs from the CAT or GLU region of the GTF of mutans streptococci or coimmunized with a combination of these constructs (CAT-GLU). Coimmunized animals demonstrated significantly higher serum immunoglobulin G (IgG) and salivary IgA antibody levels to CAT or GTF than rats immunized with either construct alone. To assess the functional significance of coimmunization with these constructs, animals were immunized as above or with Streptococcus sobrinus GTF and then infected with S. sobrinus to explore the effects of immunization on immunological, microbiological, and disease (dental caries) parameters. Serum antibody from the communized group inhibited S. sobrinus GTF-mediated insoluble glucan synthesis in vitro above that of the individual-construct-immunized groups. Immunization with CAT or GLU constructs resulted in significantly reduced dental caries after infection with S. sobrinus compared with sham-immunized animals. Coimmunization produced greater reductions in caries than after immunization with either CAT or GLU. Also, significant elevations in lymphocyte proliferative responses to CAT, GLU, and GTF were observed after coimmunization with CAT-GLU compared with the responses after immunization with the individual constructs. The results suggested that increased numbers of memory T cells, which could proliferate to CAT, were generated by coimmunization. The experiments support the functional significance of these GTF domains in dental caries pathogenesis and present coimmunization as a simple alternative to intact GTF to enhance protective immunity against cariogenic microorganisms. <23> UI - 20231798 AU - Clancy KA AU - Pearson S AU - Bowen WH AU - Burne RA IN - Department of Microbiology and Immunology and Center for Oral Biology, University of Rochester Medical Center, Rochester, New York 14642, USA. TI - Characterization of recombinant, ureolytic Streptococcus mutans demonstrates an inverse relationship between dental plaque ureolytic capacity and cariogenicity. SO - Infection & Immunity 2000 May;68(5):2621-9 AB - Dental caries results from prolonged plaque acidification that leads to the establishment of a cariogenic microflora and demineralization of the tooth. Urease enzymes of oral bacteria hydrolyze urea to ammonia, which can neutralize plaque acids. To begin to examine the relationship between plaque ureolytic activity and the incidence of dental caries, recombinant, ureolytic strains of Streptococcus mutans were constructed. Specifically, the ureABCEFGD operon from Streptococcus salivarius 57.I was integrated into the S. mutans chromosome in such a way that the operon was transcribed from a weak, cognate promoter in S. mutans ACUS4 or a stronger promoter in S. mutans ACUS6. Both strains expressed NiCl(2)-dependent urease activity, but the maximal urease levels in ACUS6 were threefold higher than those in ACUS4. In vitro pH drop experiments demonstrated that the ability of the recombinant S. mutans strains to moderate a decrease in pH during the simultaneous metabolism of glucose and urea increased proportionately with the level of urease activity expressed. Specific-pathogen-free rats that were infected with ACUS6 and fed a cariogenic diet with drinking water containing 25 mM urea and 50 microM NiCl(2) had relatively high levels of oral urease activity, as well as dramatic decreases in the prevalence of smooth-surface caries and the severity of sulcal caries, relative to controls. Urease activity appears to influence plaque biochemistry and metabolism in a manner that reduces cariogenicity, suggesting that recombinant, ureolytic bacteria may be useful to promote dental health. <24> UI - 20225498 AU - Byers MR AU - Chudler EH AU - Iadarola MJ IN - Department of Anesthesiology, University of Washington, Seattle, WA 98195-6540, USA. byersm@u.washington.edu TI - Chronic tooth pulp inflammation causes transient and persistent expression of Fos in dynorphin-rich regions of rat brainstem. SO - Brain Research 2000 Apr 10;861(2):191-207 AB - We have analyzed central Fos immunoreactivity (Fos-IR) brainstems of adult rats after three clinically relevant dental injuries: filled dentin (DF) cavities that cause mild pulp injury and heal within 1-2 weeks; open pulp exposures (PX) that cause gradual pulp loss and subsequent periodontal lesions; and filled pulp exposures (PXF). By 1 week after DF cavities, no Fos-IR remained except for sites such as lateral-ventral periolivary nucleus (LVPO) that had Fos-IR in all rats including controls. PX injury induced (1) a delayed transient expression of Fos at 1-2 weeks at three loci (ipsilateral neurons in dorsomedial nucleus oralis, paratrigeminal nucleus, and trigeminal tract), (2) persistent ipsilateral Fos for at least 4 weeks after injury in dynorphin (Dyn)-rich regions (rostral lateral solitary nucleus, periobex dorsal nucleus caudalis), and (3) late Fos-IR at 2-4 weeks (bilateral superficial cervical dorsal horn, contralateral dorsal nucleus caudalis, contralateral rostral lateral solitary nucleus). Rats with PXF injury were examined at 2 weeks, and they had greater numbers and more extensive rostro-caudal distribution of Fos neurons than the PX group. One week after PX injury, Fos-IR neurons were found in regions with strong Dyn-IR central fibers. Co-expression of Dyn and Fos was found in some unusually large neurons of the ipsilateral rostral lateral solitary nucleus, trigeminal tract, and dorsal nucleus caudalis. Immunocytochemistry for the p75 low affinity neurotrophin receptor (p75NTR) or for calcitonin gene-related peptide (CGRP) showed no consistent change in trigeminal central endings in any Fos-reactive brainstem areas, despite the extensive structural and cytochemical reorganization of the peripheral endings of the dental neurons. The Fos responses of central neurons to tooth injury have some unusual temporal and spatial patterns in adult rats compared to other trigeminal injury models. <25> UI - 20278369 AU - Lyritis GP AU - Schoenau E AU - Skarantavos G IN - Laboratory for the Research of Musculoskeletal System, University of Athens, KAT Hospital, Greece. TI - Osteopenic syndromes in the adolescent female. SO - Annals of the New York Academy of Sciences 2000;900:403-8 AB - Low bone density in growing girls and mature young women is usually a finding that needs an explanation and further clinical investigation. Population-based epidemiologic studies on osteoporosis in young persons do not exist. As a disease, osteoporosis among children and adolescents is rate, and since 1965 only 100 cases of idiopathic juvenile osteoprososis have been reported. When osteoporosis occurs in children, it is usually secondary to an underlying medical disorder (e.g., anorexia nervosa, leukemia) or to medications, but occasionally no identifiable primary cause can be detected. It may also be the result of a genetic disorder such as osteogenesis imperfecta. On the other hand, osteopenia in growing and young persons seems much commoner and needs further investigation. Adolescence is a period of increased calcium requirement, and girls with an underlying bone disease are at higher risk for bone demineralization. An additional point of interest is the changes in the geometry of bones through their continuous adaptation to simultaneous skeletal and muscular growth. Bones, through the mechanostat mechanism, adapt to mechanical loading by differentiating their geometry. A recent finding in this direction is that before and during the teenage years there is an environmental effect of physical activity and nutrition on hip geometry. Another important finding is an age-dependent increase in bone cross-sectional area and bone strength index in the absence of an increase in volumetric spongiosa bone density and cortical bone density. Girls, in comparison to boys, deposit more calcium in their bones during puberty, thus probably preparing their skeleton for the forthcoming events of pregnancy and lactation. <26> UI - 20273489 AU - Karczewska A AU - Nawrocki S AU - Breborowicz D AU - Filas V AU - Mackiewicz A IN - Department of Cancer Immunology, University School of Medical Sciences at Great Poland Cancer Center, Poznan. TI - Expression of interleukin-6, interleukin-6 receptor, and glycoprotein 130 correlates with good prognoses for patients with breast carcinoma. SO - Cancer 2000 May 1;88(9):2061-71 AB - BACKGROUND: Interleukin-6 (IL-6) is secreted by normal epithelial breast cells but not by oncogene-transformed cells. Interleukin-6 is able to inhibit growth of breast carcinoma cells in culture. Interleukin-6 exerts its activity via two receptor subunits, IL-6R and glycoprotein 130 (gp130). The expression of these receptor subunits in breast tumors has been studied, but there are no previous reports of their prognostic significance, to the authors' knowledge. METHODS: mRNA of IL-6, IL-6R, and gp130 was studied in 75 tumor samples obtained from breast carcinoma patients. Patients were followed for a maximum of 71 months (median follow-up, 61 months; 60 patients were followed for a minimum of 5 years or died during the observation period). Prognostic factors were analyzed in univariate and multivariate analysis. RESULTS: mRNA specific to IL-6, IL-6R, and gp130 was detected in 57%, 53%, and 71% of breast carcinoma tissues, respectively. Expression was strongly correlated with earlier stages of the disease. In univariate analysis, expression of IL-6 and its receptor subunits proved to be a positive prognostic factor for overall survival (OS) and disease free survival (DFS). IL-6R and gp130 expression were good independent prognostic factors for OS. The 5-year OS of all patients was 66%. The 5-year OS in IL-6, IL-6R, and gp130 positive groups was 95%, 94%, and 90%, respectively, whereas in negative groups it was 26%, 31%, and 9%, respectively. CONCLUSIONS: Expression of IL-6, IL-6R, and gp130 in breast carcinoma tissue is associated with earlier stages of the disease. In advanced stages, expression of IL-6 and its receptor subunits predicts better prognosis. <27> UI - 20206674 AU - Davidson B AU - Gotlieb WH AU - Ben-Baruch G AU - Kopolovic J AU - Goldberg I AU - Nesland JM AU - Berner A AU - Bjamer A AU - Bryne M IN - Department of Pathology, The Norwegian Radium Hospital, Oslo, Norway. bend@labmed.uio.no TI - Expression of carbohydrate antigens in advanced-stage ovarian carcinomas and their metastases-A clinicopathologic study. SO - Gynecologic Oncology 2000 Apr;77(1):35-43 AB - OBJECTIVE: Up-regulated expression or loss of expression of various carbohydrate antigens on the surface of cancer cells has been associated with a metastatic phenotype and poor survival in epithelial malignancies of different origins. The object of this study was to investigate the expression of carbohydrate antigens in two groups of patients diagnosed with advanced-stage ovarian carcinoma-one with an extremely favorable outcome and the other with a uniformly poor survival. METHODS: Sections from 76 paraffin-embedded blocks (primary ovarian carcinomas and metastatic lesions) from 45 patients diagnosed with advanced-stage ovarian carcinomas (FIGO stages III-IV) were immunohistochemically stained using five monoclonal antibodies for Lewis(y) (Le(y))(two antibodies), Sialyl Lewis(x) (Slex), Tn, and Sialyl Tn (STn) antigens. Patients were divided in two groups based on outcome. Long-term survivors (21 patients) and short-term survivors (24 patients) were defined using a double cut-off of 36 months for disease-free survival (DFS) and 60 months for overall survival (OS). Staining results for primary tumors and metastases were analyzed separately. RESULTS: Mean follow-up period was 70 months. The mean values for DFS and OS were 109 and 125 months for long-term survivors and 3 and 25 months for short-term survivors. Staining for all four antigens was seen in the majority of cases (range = 72-96%) and tended to be comparable in primary tumors and their metastases. However, absence of immunoreactivity for STn was seen in 9/38 (24%) metastatic lesions and only 1/38 (3%) primary tumors. This finding did not reach statistical significance (P > 0.05). A combined pattern of membranous and cytoplasmic staining was predominant in the majority of cases. Enhanced staining for Le(y) and STn was detected in the invasive front of some tumors, while Slex and Tn immunoreactivity did not relate to cell location. Primary tumors and metastatic lesions of long-term survivors displayed immunoreactivity patterns that were comparable to those of short-term survivors. In the evaluation of survival curves, more diffuse staining for Slex showed marginal correlation with poor survival (P = 0.05), while a trend toward poorer survival was seen in tumors that were more extensively stained for Le(y) and Tn (P > 0.05). CONCLUSIONS: Le(y), Slex, STn, and Tn antigens are widely expressed in primary ovarian carcinomas and their metastases. Altered expression of Sialyl Tn is observed with tumor progression in a fraction of ovarian carcinomas. Expression of membrane carbohydrate residues is prevalent in tumors of both long-term and short-term survivors and does not appear to be a strong predictor of disease outcome. However, larger studies are needed to further elucidate the role of these molecules in ovarian carcinogenesis. Copyright 2000 Academic Press. <28> UI - 20252596 AU - Eissa S AU - Kassim SK AU - Imam M AU - Khalifa A IN - Biochemistry Department, Ain Shams Faculty of Medicine, Abbassia, Cairo, Egypt. samar:kassim@usa.net TI - Correlation between EBV DNA and rearrangement and expression of Bcl-2 gene in aggressive non-Hodgkin's lymphoma. SO - Iubmb Life 1999 Aug;48(2):231-6 AB - Previous in vitro studies have shown that bcl-2 expression can be induced by transfection of Epstein-Barr virus (EBV)-negative non-Hodgkin's lymphoma (NHL) cell lines with EBV. This induced expression of bcl-2 is important for the long survival of EBV-positive cells and might be a first step in tumorigenesis. The purpose of the present study was to investigate the possibility of similar correlation between bcl-2 expression and EBV infection in vivo in a cohort of patients with aggressive NHL, who were uniformly evaluated and treated with effective chemotherapy. The 42 patients included were 25-65 years old. None had prior treatment, discordant lymphoma, or human immunodeficiency virus seropositivity. Fresh biopsied samples were obtained and stored frozen for analysis of bcl-2 gene rearrangement major break point and of EBV DNA by PCR. Bcl-2 protein expression was estimated by Western blot, and enzyme immunoassay. With a median follow-up of 30 months, overall survival (OS) and disease-free survival (DFS) were measured to determine the prognostic significance of these variables. Analyzable DNA was present in all samples, 24% demonstrating bcl-2 rearrangement and 33% showing EBV DNA. Patients with bcl-2 gene rearrangement tended to have shorter DFS, and OS than patients without translocation. Bcl-2 protein expression was not correlated to gene rearrangement and had no significant influence on survival. The presence of EBV DNA in NHL had no prognostic significance but was correlated to bcl-2 expression. EBV-positive tumors showed higher bcl-2 expression than EBV-negative tumors did. Our results suggest a role of EBV infection in inducing bcl-2 expression as a survival factor for EBV-positive cells. <29> UI - 20244199 AU - Hsu YL AU - Wang KH AU - Yang YH AU - Tung MC AU - Hu CH AU - Lo CF AU - Wang CH AU - Hsu T IN - Institute of Zoology, Academia Sinica, Taipei, Taiwan, ROC. TI - Diagnosis of Penaeus monodon-type baculovirus by PCR and by ELISA of occlusion bodies. SO - Diseases of Aquatic Organisms 2000 Mar 14;40(2):93-9 AB - The black tiger prawn Penaeus monodon is a valuable aquaculture product in Taiwan. Two specific diagnostic methods were established for P. monodon-type baculovirus, one using polymerase chain reaction (PCR) technology and the other enzyme-linked immunosorbent assay (ELISA) technology. Monodon-type baculovirus (MBV) was purified by sucrose gradient centrifugation from occlusion bodies of MBV-infected postlarvae of P. monodon. MBV DNA was subsequently purified from the occlusion bodies and its presence was confirmed by PCR using primers of the polyhedrin gene. Based on conserved sequences of the DNA polymerase genes of Autographa californica nuclear polyhedrosis virus (AcMNPV) and Lymantria dispar nuclear polyhedrosis virus (LdMNPV), primers were designed and synthesized to yield a 714 bp PCR fragment from MBV. However, the sequence of this fragment revealed low homology with that of LdMNPV and AcMNPV. From the DNA sequence of this fragment, a second set of primers was designed, and using these primers, a 511 bp DNA fragment was amplified only when MBV DNA was the template. DNA templates from AcMNPV, white spot syndrome diseased shrimp, or PMO cells (a cell line derived from the Oka organ of Penaeus monodon) did not give any amplified DNA fragment. Therefore, this primer pair was specific for the diagnosis of MBV. By using intraspleenic immunization of rabbits with purified MBV occlusion bodies, a polyclonal rabbit antiserum against MBV was obtained. This antiserum could detect nanogram levels of MBV, but did not cross react with white spot syndrome virus (WSSV), homogenates of PMO cells, postlarvae, hepatopancreatic tissue or intestinal tissue of black tiger prawns by competitive ELISA. This sensitive method could detect MBV even in tissue homogenates. <30> UI - 20203507 AU - Woodnutt DA AU - Wager-Miller J AU - O'Neill PC AU - Bothwell M AU - Byers MR IN - Dental School, University of Washington, Seattle, USA. TI - Neurotrophin receptors and nerve growth factor are differentially expressed in adjacent nonneuronal cells of normal and injured tooth pulp. SO - Cell & Tissue Research 2000 Feb;299(2):225-36 AB - High-affinity tyrosine kinase A (trkA) neurotrophin receptors on neurons and nonneuronal cells elicit differentiation or survival functions in response to nerve growth factor (NGF), whereas the low-affinity neurotrophin (p75) receptor modulates trkA activity or can independently cause apoptosis or NFkappaB-mediated survival functions. We examined dental tissues for the presence of trkA-like immunoreactivity (trkA-IR), to determine which nonneuronal cell types express it in normal compared with inflamed teeth and how the trkA-positive cells relate to those expressing the p75 receptor and/or NGF. Normal and injured rat molars (dentin cavity for 4 h, 16-24 h, 3 days, 16 days, or 5 weeks) were immunoreacted using the ABC detection system for two anti-trkA antibodies (sTA, Santa Cruz Biotechnology; rTA, L. Reichardt) and antibodies against p75 and NGF, all of which also stained pulpal nerve fibers. We report that, when using the sTA antibody (recognizing the intracellular carboxy terminal), nonneuronal trkA-IR was found in odontoblasts of normal teeth and also in invading polymorphonuclear leukocytes (PMNs) and reparative odontoblasts after injury. When using rTA (recognizing the extracellular domain of the receptor), nonneuronal trkA-IR was only found in odontoblasts. Odontoblasts also had NGF-IR but did not label for NGF mRNA. The lack of odontoblast NGF mRNA suggests that NGF is passed from fibroblasts to the adjacent odontoblasts, where it is picked up by receptor-mediated mechanisms for regulation of odontoblast function. Tooth injury disrupts this system such that trkA-IR decreases in injured odontoblasts, p75 decreases in fibroblasts, and NGF is upregulated by fibroblasts and accumulates in the injured pulp and surviving odontoblasts. Pulpal NGF may contribute to chemoattraction for the invading leukocytes or their sTA-IR may have been induced in response to pulpal NGF. Thus, tooth pulp has a different distribution of nonneuronal NGF and its paracrine receptors during inflammation compared with normal conditions. <31> UI - 20228595 AU - Ma JK IN - Department of Oral Medicine and Pathology, Guy's Hospital, London. TI - The caries vaccine: a growing prospect. SO - Dental Update 1999 Nov;26(9):374-80 AB - A vaccine against dental caries, for so long a subject of purely academic research, is currently undergoing phase II clinical trials and could be available commercially within 5 to 6 years. The approach, which is safe, effective, and provides long-term protection for up to a year, is based on a topical application and does not require any injections. The development of this vaccine has been made possible by recent advances in molecular biology and genetic engineering. Perhaps one of the most intriguing aspects has been the use of green plants to produce the vaccine. This article describes a topical vaccine against Streptococcus mutans, which is the main cause of dental caries. <32> UI - 20214217 AU - Wang Q AU - Poulos BT AU - Lightner DV IN - Max-Planck-Institute for Biology, Tuebingen, Germany. TI - Protein analysis of geographic isolates of shrimp white spot syndrome virus. SO - Archives of Virology 2000;145(2):263-74 AB - Six geographic isolates of the white spot syndrome virus (WSSV) of penaeid shrimp, from China, India, Thailand, South Carolina, Texas, as well as from crayfish kept at the US National Zoo in Washington D. C, were compared by electron microscopy and sodium sulfate polyacrylamine gel electrophoresis (SDS-PAGE). Amino acid compositions of four of the major structural polypeptides of the South Carolina WSSV were analyzed, and three of the four polypeptides were partially sequenced from their NH2 termini. The morphologies of purified virions of the six geographic isolates of WSSV were indistinguishable by transmission electron microscopy. By SDS-PAGE, the protein profiles of the six isolates were very similar, but not identical. They all contained three major polypeptides with sizes of approximately 25, 23 and 19 kDa. A fourth major polypeptide at the 14.5 kDa position was observed in four of the geographic isolates. The WSSV isolated from crayfish presented a slightly different structural protein profile, particularly with regard to the protein in the 19 kDa range that appeared larger in size than those of the other isolates. The NH2 terminal amino acids of the 25, 23 and 14.5 kDa polypeptides of the South Carolina WSSV were sequenced as MDLSFTLSVVTA, MEFGNLTNLDVA, and VARGGKTKGRRG, respectively. No significant homologous sequence was found in the GenBank. These protein sequences have been submitted to the SWISS-PROT Protein Data Bank and assigned the accession numbers P82004, P82005 and P82006. <33> UI - 20196979 AU - Talwar GP AU - Diwan M AU - Razvi F AU - Malhotra R IN - Talwar Research Foundation, New Delhi, India. TI - The impact of new technologies on vaccines. [Review] [75 refs] SO - National Medical Journal of India 1999 Nov-Dec;12(6):274-80 AB - Vast changes are taking place in vaccinology consequent to the introduction of new technologies. Amongst the vaccines included in the Expanded Programme of Immunization (EPI), the pertussis vaccine has been replaced by acellular purified fractions devoid of side-effects. Non-pathogenic but immunogenic mutants of tetanus and diptheria toxins are likely to replace the toxoids. An effective vaccine against hepatitis B prepared by recombinant technology is in large-scale use. Conjugated vaccines against Haemophilus influenzae b, S. pneumococcus and meningococcus are now available, as also vaccines against mumps, rubella and measles. Combination vaccines have been devised to limit the number of injections. Vaccine delivery systems have been developed to deliver multiple doses of the vaccine at a single contact point. A genetically-engineered oral vaccine for typhoid imparts better and longer duration of immunity. Oral vaccines for cholera and other enteric infections are under clinical trials. The nose as a route for immunization is showing promise for mucosal immunity and for anti-inflammatory experimental vaccines against multiple sclerosis and insulin-dependent diabetes mellitus. The range of vaccines has expanded to include pathogens resident in the body such as Helicobacter pylori (duodenal ulcer), S. mutans (dental caries), and human papilloma virus (carcinoma of the cervix). An important progress is the recognition that DNA alone can constitute the vaccines, inducing both humoral and cell-mediated immune responses. A large number of DNA vaccines have been made and shown interesting results in experimental animals. Live recombinant vaccines against rabies and rinderpest have proven to be highly effective for controlling these infections in the field, and those for AIDS are under clinical trial. Potent adjuvants have added to the efficacy of the vaccines. New technologies have emerged to 'humanize' mouse monoclonals by genetic engineering and express these efficiently in plants. These recombinant antibodies are opening out an era of highly specific and safe therapeutic interventions. Human recombinant antibodies would be invaluable for treating patients with terminal tetanus and rabies. Antibodies are already in use for treatment of cancer, rheumatoid arthritis and allergies. An advantage of preformed antibodies directed at a defined target and given in adequate amounts is the certainty of efficacy in every recipient, in contrast to vaccines, where the quality and quantum of immune response varies from individual to individual. [References: 75] <34> UI - 20197555 AU - Ito Y AU - Monden M AU - Takeda T AU - Eguchi H AU - Umeshita K AU - Nagano H AU - Nakamori S AU - Dono K AU - Sakon M AU - Nakamura M AU - Tsujimoto M AU - Nakahara M AU - Nakao K AU - Yokosaki Y AU - Matsuura N IN - Department of Surgery II, Osaka University Medical School, Japan. TI - The status of Fas and Fas ligand expression can predict recurrence of hepatocellular carcinoma. SO - British Journal of Cancer 2000 Mar;82(6):1211-7 AB - The status of Fas and Fas ligand (Fas L) expression was investigated in this study for 103 hepatocellular carcinomas (HCC). We studied the expression of the following three factors, Fas and Fas L expression in carcinoma cells and Fas L expression in stromal mononuclear cells (defined as stromal Fas L index). Fas expression in HCC cells was significantly decreased in cases with poor differentiation (P < 0.0001) and of larger size (P = 0.0058). Fas L expression in carcinoma cells was observed exclusively in moderately or poorly differentiated cases. Furthermore, each factor had prognostic significance for disease-free survival (DFS) (P< 0.0001, P = 0.0222 and 0.0027 respectively). We then scored the results of each factor and defined the total score as 'Fas-Fas L risk score'. The P-value of the score for DFS was even lower than that of the clinical stage by multivariate analysis. These results suggest that the evaluation of Fas and Fas ligand expression potentially has a significant prognostic value for DFS of HCC patients, in addition to the clinical stage, and can be regarded as a new prognostic marker. <35> UI - 20141334 AU - Hoshino T AU - Mizuta K AU - Gao J AU - Araki S AU - Araki K AU - Takeshita T AU - Wu R AU - Morita H IN - Department of Otolaryngology, Hamamatsu University School of Medicine, 3600 Handa-cho, Hamamatsu, Japan. thoshino@hama-med.ac.jp TI - Cochlear findings in the white spotting (Ws) rat. SO - Hearing Research 2000 Feb;140(1-2):145-56 AB - White spotting (Ws) rats possess a c-kit gene mutation at the W locus, resulting in a variety of characteristics including a lack of intermediate cells of the stria vascularis. The present study employs a light microscope (LM), scanning (SEM) and transmission electron microscopes (TEM), diaminobenzidine (DAB) staining techniques and auditory brainstem response (ABR) to investigate the structure and function of the cochlea in 26 homozygous Ws/Ws rats aged 1-6 months. A slight thinning of the stria vascularis and moderate elevation of ABR threshold were about the only defects noted in 1 month animals, while older animals displayed various defects that tended to worsen with age. At 3 months LM revealed pigment granules in the basal turn of most animals, with a loss of pigmentation in the upper turns. The stria vascularis and organ of Corti tended to be well preserved in the lower, pigmented portion, while the upper, unpigmented portion showed severe strial degeneration and some outer hair cell loss. DAB staining revealed a well developed strial capillary net throughout the pigmented portion of the cochlea, with severe degradation in the unpigmented apical portion. ABR thresholds were slightly elevated over 1 month values. At 6 months great differences in degeneration were noted between right and left ears of the same animal. <36> UI - 20178632 AU - Martin G AU - Halwani F AU - Shibata H AU - Meterissian S IN - Department of Surgical Oncology, Royal Victoria Hospital, McGill University, Montreal, Que. TI - Value of DNA ploidy and S-phase fraction as prognostic factors in stage III cutaneous melanoma. SO - Canadian Journal of Surgery 2000 Feb;43(1):29-34 AB - OBJECTIVE: To determine the prognostic value of flow cytometric analysis (S-phase fraction and DNA index) performed on lymph-node metastases of patients with stage III melanoma. DESIGN: A retrospective chart review with flow cytometric analysis of paraffin-embedded tissues. SETTING: A university teaching hospital. PATIENTS: Among 332 patients with cutaneous melanoma, 33 with stage III were identified. Distant metastases developed in 16 patients; 17 had no further recurrence. Charts were reviewed to obtain clinicopathologic parameters such as sex, age, location of the primary tumour, histologic features, presence or absence of ulceration, and Clark's and Breslow's levels. INTERVENTION: DNA ploidy and S-phase fraction were determined on the paraffin-embedded nodes. MAIN OUTCOME MEASURES: The groups with or without recurrence were compared in terms of disease-free survival (DFS) and overall survival (OS). These survival parameters were correlated with DNA ploidy and S-phase fraction. RESULTS: By univariate analysis, clinicopathologic factors did not predict OS. A higher Clark's level of invasion and more than 3 positive lymph nodes were associated with shorter DFS (p < 0.05). Tumour thickness and S-phase fraction did not correlate with either DFS or OS. Patients with diploid lymph-node metastases had an 87% 12-month survival compared with 41% for those with aneuploid tumours. CONCLUSIONS: DNA ploidy may be used as a prognostic index in patients with lymph-node metastases. This could be particularly useful in the context of sentinel lymph-node mapping by which more patients are being identified with single microscopic lymph-node involvement. <37> UI - 20113111 AU - Rhim H AU - Dunn KJ AU - Aronzon A AU - Mac S AU - Cheng M AU - Lamoreux ML AU - Tilghman SM AU - Pavan WJ IN - Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health (NIH), Bethesda, Maryland 20892-4472 USA. TI - Spatially restricted hypopigmentation associated with an Ednrbs-modifying locus on mouse chromosome 10. SO - Genome Research 2000 Jan;10(1):17-29 AB - We have used the varied expressivity of white spotting (hypopigmentation) observed in intrasubspecific crosses of Ednrb(s) mice (Mayer Ednrb(s)/Ednrb(s) and C3HeB/FeJ Ednrb(s)/Ednrb(s)) to analyze the effects of modifier loci on the patterning of hypopigmentation. We have confirmed that an Ednrb(s) modifier locus is present on mouse Chromosome 10. This locus is now termed k10, using the nomenclature established by Dunn in 1920. The k10(Mayer) allele is a recessive modifier that accounts for almost all of the genetic variance of dorsal hypopigmentation. Using intercross analyses we identified a second allele of this locus or a closely linked gene termed k10(C3H). The k10(C3H) allele is semidominant and is associated with the penetrance and expressivity of a white forelock phenotype similar to that seen in Waardenburg syndrome. Molecular linkage analysis was used to determine that the k10 critical interval was flanked by D10Mit10 and D10Mit162/D10Mit122 and cosegregates with mast cell growth factor (Mgf). Complementation crosses with a Mgf(Sl) allele (a 3-5-cM deletion) confirm the semidominant white forelock feature of the k10(C3H) allele and the dorsal spotting feature of K10(Mayer) allele. MgF was assessed as a candidate gene for k10(Mayer) and k10(C3H) by sequence and genomic analyses. No molecular differences were observed between the Mayer and C57BL/6J alleles of MgF; however, extensive genomic differences were observed between the C3HeB/FeJ and C57BL/6J alleles. This suggests that alteration of MgF expression in C3H mice may account for the k10(C3H) action on white forelock hypopigmentation. Crosses of Ednrb(s) with Kit(WJ-2) (the receptor for MGF)-deficient mice confirmed the hypothesis that synergistic interaction between the Endothelin and MGF signaling pathways regulates proper neural crest-derived melanocyte development in vivo. <38> UI - 20115090 AU - Liu B AU - Rayment SA AU - Gyurko C AU - Oppenheim FG AU - Offner GD AU - Troxler RF IN - Department of Periodontology and Oral Biology, Goldman School of Graduate Dentistry, Boston University School of Medicine, Boston University Medical Center, MA 02118, Boston, USA. TI - The recombinant N-terminal region of human salivary mucin MG2 (MUC7) contains a binding domain for oral Streptococci and exhibits candidacidal activity. SO - Biochemical Journal 2000 Feb 1;345 Pt 3:557-64 AB - MG2 (the MUC7 gene product) is a low-molecular-mass mucin found in human submandibular/sublingual secretions. This mucin is believed to agglutinate a variety of microbes and thus is considered an important component of the non-immune host defence system in the oral cavity. We have shown that MUC7 can bind to cariogenic strains of Streptococcus mutans and that this binding requires a structural determinant in the N-terminal region. In the present study an expression construct, pNMuc7, encoding the N-terminal 144 amino acids of MUC7 was generated, and the recombinant protein rNMUC7 was expressed in Escherichia coli. Purified rNMUC7 was characterized and the binding of this protein to oral bacteria was investigated in an established assay. The results showed that the recombinant protein bound to S. mutans ATCC 25175 and ATCC 33402, and that alkylation of the two cysteine residues (Cys(45) and Cys(50)) resulted in the complete loss of bacterial binding. This suggests that binding of MUC7 to S. mutans occurs between the N-terminal region of the mucin molecule and the bacterial surface, and that this interaction is dependent on a cysteine-containing domain within this region of MUC7. In addition, the killing activity of rNMUC7 was compared with that of the candidacidal salivary protein histatin 5 in an established Candida albicans (ATCC 44505) blastoconidia killing assay. It was found that the LD(50) values of rNMUC7 and histatin 5 were comparable, and that the recombinant protein displayed significant killing activity at the physiological concentration range of MUC7 in whole saliva. This study is the first to show that the N-terminal region of MUC7 contains a structural determinant for bacterial binding and that this region exhibits candidacidal activity. <39> UI - 20168644 AU - Cuny M AU - Kramar A AU - Courjal F AU - Johannsdottir V AU - Iacopetta B AU - Fontaine H AU - Grenier J AU - Culine S AU - Theillet C IN - Genome et Cancer UMR 5535 Centre National de la Recherche Scientifique, Centre de Recherche et de Lutte contre le Cancer Val d'Aurelle-Paul Lamarque, Montpellier, France. TI - Relating genotype and phenotype in breast cancer: an analysis of the prognostic significance of amplification at eight different genes or loci and of p53 mutations. SO - Cancer Research 2000 Feb 15;60(4):1077-83 AB - Breast cancer heterogeneity can be related directly to its variability at the genetic level. Thus, tumor genotyping could be a valuable approach to define breast tumor subtypes. It has been shown that it is possible to delineate subgroups of breast tumors according to specific sets of DNA amplifications. The aim of the present work was to study the prognostic significance of these DNA amplifications. We studied DNA amplification at eight genes or loci (AIB1, CCND1, EMS1, ERBB2, FGFR1, MDM2, MYC, and RMC20C001) as well as p53 mutations in a series of 640 breast cancer patients who had not received presurgical therapy and analyzed the correlations with survival DNA amplification was assessed by Southern blotting and was scored positive when exceeding three to five copies. Mutations in the p53 gene were searched by four-color fluorescent single. strand conformational polymorphism, using an automated sequencer. Of the nine genetic alterations tested, four (CCND1, EMS1, FGFR1, and p53 mutations) showed a significant association with reduced disease-free (DFS) and/or overall survival (OVS) in the unselected set of patients by univariate test. Correlations for p53 were found only when selecting mutations in exons 5 or 7. Analysis of node-negative and -positive subgroups of patients showed that MDM2 amplification and p53 mutations bore prognostic significance in node-negative patients, whereas amplification of CCND1, EMS1, and FGFR1 correlated with poor outcome in node-positive patients. Multivariate analysis on an unselected set of patients retained significance for the amplification of EMS1, FGFR1, and MDM2 with DFS, of CCND1 with OVS, and of RMC20C001 with both DFS and OVS. Interestingly, stratified analysis according to nodal status confirmed results obtained in the univariate tests: significance of MDM2 amplification and p53 mutations in node-negative and that of CCND1, EMS1, and FGFR1 in node-positive patients. We also observed an association between the number of genetic alterations observed in a tumor and poor prognosis. Patients with two or more amplified loci had a worsened outcome. Strongly correlating coamplifications such as CCND1 and FGFR1, as well as ERBB2 and MYC, were associated with a significant reduction of patient survival, thus indicating cooperative effects. Our data support the idea that genetic alterations in breast cancer are not only helpful for phenotyping purposes, but can also represent powerful prognostic indicators in the clinical practice. <40> UI - 20128227 AU - Hasegawa Y AU - Tokuyama T AU - Iwaki H IN - Department of Biotechnology, Faculty of Engineering, Kansai University, Osaka, Japan. yoshie@ipcku.kansai-u.ac.jp TI - Cloning and expression of the N,N-dimethylformamidase gene from Alcaligenes sp. strain KUFA-1. SO - Bioscience, Biotechnology & Biochemistry 1999 Dec;63(12):2091-6 AB - N,N-Dimethylformamidase (DMFase) from Alcaligenes sp. strain KUFA-1, a bacterium that can grow on N,N-dimethylformamide (DMF) as the sole carbon and nitrogen source, catalyzes the first step of the DMF degradation. The DMFase gene dmfA1A2 was cloned in Escherichia coli, and its nucleotides were sequenced. The deduced amino acid sequence of the enzyme consisted of two alpha- and two beta-subunits with 132 and 762 amino acids, respectively, and had little similarity to sequences in protein databases, including various amidases. The protein may be a new kind of amidase. DMFase activity was detected in E. coli cells transformed with an expression plasmid of the cloned DMFase gene. The properties of recombinant DMFase purified from E. coli were identical to those of Alcaligenes DMFase. <41> UI - 20120090 AU - Poole LB AU - Higuchi M AU - Shimada M AU - Calzi ML AU - Kamio Y IN - Department of Biochemistry, Wake Forest University School of Medicine, Winston-Salem 27157, NC, USA. lbpoole@wfubmc.edu TI - Streptococcus mutans H2O2-forming NADH oxidase is an alkyl hydroperoxide reductase protein. SO - Free Radical Biology & Medicine 2000 Jan 1;28(1):108-20 AB - Nox-1 from Streptococcus mutans, the bacteria which cause dental caries, was previously identified as an H2O2-forming reduced nicotinamide adenine dinucleotide (NADH) oxidase. Nox-1 is homologous with the flavoprotein component, AhpF, of Salmonella typhimurium alkyl hydroperoxide reductase. A partial open reading frame upstream of nox1, homologous with the other (peroxidase) component, ahpC, from the S. typhimurium system, was also identified. We report here the complete sequence of S. mutans ahpC. Analyses of purified AhpC together with Nox-1 have verified that these proteins act as a cysteine-based peroxidase system in S. mutans, catalyzing the NADH-dependent reduction of organic hydroperoxides or H2O2 to their respective alcohols and/or H2O. These proteins also catalyze the four-electron reduction of O2 to H2O2, clarifying the role of Nox-1 as a protective protein against oxygen toxicity. Major differences between Nox-1 and AhpF include: (i) the absolute specificity of Nox-1 for NADH; (ii) lower amounts of flavin semiquinone and a more prominent FADH2 to NAD+ charge transfer absorbance band stabilized by Nox-1; and (iii) even higher redox potentials of disulfide centers relative to flavin for Nox-1. Although Nox-1 and AhpC from S. mutans were shown to play a protective role against oxidative stress in vitro and in vivo in Escherichia coli, the lack of a significant effect on deletion of these genes from S. mutans suggests the presence of additional antioxidant proteins in these bacteria. <42> UI - 20121580 AU - Mochizuki K AU - Yonezu T AU - Yakushiji M AU - Machida Y IN - Department of Pediatric Dentistry, Tokyo Dental College, Chiba, Japan. TI - The fusion of three primary incisors: report of case. [Review] [26 refs] SO - ASDC Journal of Dentistry for Children 1999 Nov-Dec;66(6):421-5, 367 AB - The occurrence of a fusion of three primary incisors is rare. A two-year-old Japanese girl was brought to the pediatric dental outpatient clinic, Tokyo Dental College, to receive a caries-prevention treatment. The fused tooth consisted of the maxillary primary central incisors and right maxillary lateral incisor. Primary left lateral incisor erupted normally and the other primary teeth were erupted. The radicular pulp chambers were fused into one for three fourths of the length from the apex, and only one root. The hair and skin of the patient appeared normal and no systemic abnormality or congenital disease was noted in the medical history of the patient and her family. The occurrence of a three-tooth fusion, and no supernumerary tooth was confirmed. [References: 26] <43> UI - 20043984 AU - Huang CC AU - Song YL IN - Department of Zoology, National Taiwan University, Taipei. TI - Maternal transmission of immunity to white spot syndrome associated virus (WSSV) in shrimp (Penaeus monodon). SO - Developmental & Comparative Immunology 1999 Oct-Dec;23(7-8):545-52 AB - Beta-1,3-1,6-glucan, derived from bakers' yeast Saccharomyces cerevisiae, was used in the present study to investigate the extent to which glucan is able to protect spawners from white spot syndrome associated virus (WSSV), and whether this protection (if any) can be passed on to hatchlings via maternal transmission of immunity. Results showed that fewer spawners in the glucan-injected groups showed the clinical symptoms of red body coloration and white spots on the shell during the 15 days between eyestalk ablation and the end of repeated spawning. This suggests that the application of glucan might lead to a slight enhancement of disease resistance in spawners, although the differences were not statistically significant within the confidence limit chosen. Challenge results showed a significant increase in relative percent survival for larvae derived from groups of glucan-injected spawners compared to those derived from groups of untreated spawners. It therefore seems that a maternally transmitted disease resistance induced by glucan, protected the larvae against a WSSV infection. Glucan immersion was not only shown to be effective for nauplii derived from spawners that were not injected with glucan, it also provided additional, cumulative protection for nauplii which already had a maternally transmitted resistance to WSSV. This is the first documented demonstration of a maternal transmission of immunity in invertebrates. <44> UI - 20107276 AU - van Hulten MC AU - Westenberg M AU - Goodall SD AU - Vlak JM IN - Laboratory of Virology, Wageningen Agricultural University, Binnenhaven 11, Wageningen, 6709 PD, The Netherlands. TI - Identification of two major virion protein genes of white spot syndrome virus of shrimp. SO - Virology 2000 Jan 20;266(2):227-36 AB - White Spot Syndrome Virus (WSSV) is an invertebrate virus, causing considerable mortality in shrimp. Two structural proteins of WSSV were identified. WSSV virions are enveloped nucleocapsids with a bacilliform morphology with an approximate size of 275 x 120 nm, and a tail-like extension at one end. The double-stranded viral DNA has an approximate size 290 kb. WSSV virions, isolated from infected shrimps, contained four major proteins: 28 kDa (VP28), 26 kDa (VP26), 24 kDa (VP24), and 19 kDa (VP19) in size, respectively. VP26 and VP24 were found associated with nucleocapsids; the others were associated with the envelope. N-terminal amino acid sequences of nucleocapsid protein VP26 and the envelope protein VP28 were obtained by protein sequencing and used to identify the respective genes (vp26 and vp28) in the WSSV genome. To confirm that the open reading frames of WSSV vp26 (612) and vp28 (612) are coding for the putative major virion proteins, they were expressed in insect cells using baculovirus vectors and analyzed by Western analysis. A polyclonal antiserum against total WSSV virions confirmed the virion origin of VP26 and VP28. Both proteins contained a putative transmembrane domain at their N terminus and many putative N- and O-glycosylation sites. These major viral proteins showed no homology to baculovirus structural proteins, suggesting, together with the lack of DNA sequence homology to other viruses, that WSSV may be a representative of a new virus family, Whispoviridae. Copyright 2000 Academic Press. <45> UI - 20112891 AU - van Hulten MC AU - Tsai MF AU - Schipper CA AU - Lo CF AU - Kou GH AU - Vlak JM IN - Laboratory of Virology, Wageningen University, Binnenhaven 11, 6709 PD Wageningen, The Netherlands. TI - Analysis of a genomic segment of white spot syndrome virus of shrimp containing ribonucleotide reductase genes and repeat regions. SO - Journal of General Virology 2000 Feb;81 Pt 2:307-16 AB - White spot syndrome is a worldwide disease of penaeid shrimp. The disease agent is a bacilliform, enveloped virus, white spot syndrome virus (WSSV), with a double-stranded DNA genome that probably contains well over 200 kb. Analysis of a 12.3 kb segment of WSSV DNA revealed eight open reading frames (ORFs), including the genes for the large (RR1) and small (RR2) subunits of ribonucleotide reductase. The rr1 and rr2 genes were separated by 5760 bp, containing several putative ORFs and two domains with multiple sequence repeats. The first domain contained six direct repeats of 54 bp and is part of a coding region. The second domain had one partial and two complete direct repeats of 253 bp at an intergenic location. This repeat, located immediately upstream of rr1, has homologues at several other locations on the WSSV genome. Phylogenetic analysis of RR1 and RR2 indicated that WSSV belongs to the eukaryotic branch of an unrooted parsimonious tree and, further, seems to suggest that WSSV and baculoviruses probably do not share an immediate common ancestor. The present analysis of WSSV favours the view that this virus is either a member of a new genus (Whispovirus) within the Baculoviridae or a member of an entirely new virus family. <46> UI - 20152144 AU - Ohishi M AU - Horibe M AU - Ikedo D AU - Miyazaki M AU - Ohishi K AU - Kataoka M AU - Kido J AU - Nagata T IN - Department of Periodontology and Endodontology, Tokushima University School of Dentistry, Japan. TI - Effect of retinoic acid on osteopontin expression in rat clonal dental pulp cells. SO - Journal of Endodontics 1999 Oct;25(10):683-5 AB - We studied the effect of retinoic acid on osteopontin synthesis and the mRNA expression in rat clonal dental pulp cells, RPC-C2A. An immunoprecipitation assay clarified that retinoic acid caused an increase in phosphorylated osteopontin synthesis that was dose-dependent, and marked increases were observed at retinoic acid concentrations of 10(-6) to 10(-5) M (1.7-fold). A Northern blotting analysis revealed a similar pattern of increase in osteopontin mRNA expression of up to 6.2-fold of control levels. Because osteopontin has an important role in the mineralization process, these results suggest that retinoic acid regulates mineralization, which takes place in the pulp cavity, including reparative dentin formation. <47> UI - 20123526 AU - Chen TC AU - Kuo T AU - Chan HL IN - Department of Pathology, Chang Gung University School of Medicine and Chang Gung Memorial Hospital, Kwei San, Tao Yuan, Taiwan. TI - Dermatofibroma is a clonal proliferative disease. SO - Journal of Cutaneous Pathology 2000 Jan;27(1):36-9 AB - Benign fibrous histiocytoma of the skin or dermatofibroma (DF) has been regarded as a fibrohistiocytic tumor. Whether DF is a neoplastic growth or a reactive process has not been settled. Since a neoplastic process is clonal in nature, clonal analysis of DF was conducted to see if DF is a clonal disease. Fresh specimens of 13 DFs and 2 hypertrophic scars obtained from female patients were studied. The adjacent nonlesional skin tissues served as controls. The clonal analysis was based on the methylation pattern of the polymorphic X-chromosome linked androgen-receptor gene (HUMARA). Eight DFs and 1 hypertrophic scar were heterozygous at the androgen receptor gene and could be analyzed. All 8 informative DFs showed a significant reduction in one of the allelic bands compared with the corresponding bands of the nonlesional tissue after Hha I digestion. Therefore, DF is a clonal proliferative disease. In contrast, 1 hypertrophic scar showed a polyclonal pattern of X-chromosome inactivation. We conclude that DF is a clonal disease favoring a neoplastic process. <48> UI - 20152089 AU - Wynn RL AU - Meiller TF AU - Crossley HL IN - Department of Oral Medicine and Diagnostic Sciences, Dental School, University of Maryland at Baltimore, USA. TI - Tobacco "plantibodies" for caries prevention. [Review] [25 refs] SO - General Dentistry 1999 Sep-Oct;47(5):450-4 <49> UI - 20137363 AU - Faderl S AU - Kantarjian HM AU - Thomas DA AU - Cortes J AU - Giles F AU - Pierce S AU - Albitar M AU - Estrov Z IN - Department of Leukemia, The University of Texas M. D. Anderson Cancer Center, Houston 77030, USA. TI - Outcome of Philadelphia chromosome-positive adult acute lymphoblastic leukemia. SO - Leukemia & Lymphoma 2000 Jan;36(3-4):263-73 AB - Philadelphia chromosome (Ph)-positive acute lymphoblastic leukemia (ALL) represents the most common cytogenetic abnormality in adult ALL. It is found in 15% to 30% of patients, and its incidence increases with age. As in children, prognosis in Ph-positive adult ALL is poor. No therapeutic approach has had substantial impact on its unfavorable course. We analyzed the characteristics and outcome of newly diagnosed adults with Ph-positive ALL treated at the M. D. Anderson Cancer Center between 1980 and 1997. The diagnosis of patients was based on typical morphological and immunophenotypic criteria of marrow aspirate and biopsy specimens. Cytogenetic and molecular studies were also performed. A total of 67 patients were included in this study. From 1980 until 1991, 38 patients with Ph-positive ALL were treated with vincristine, Adriamycin, and dexamethasone (VAD), or with acute myeloid leukemia (AML)-like induction protocols. Since 1992 a total of 29 patients received induction therapy with an intensified treatment protocol, called "hyper-CVAD". The outcome of patients treated with standard and intensified treatment regimens was compared and results of our institution contrasted with data obtained from other centers. Ph-positive ALL was present in 67 of 498 patients with newly diagnosed ALL (13%). Patients with Ph-positive ALL had a higher median age (44 versus 34, P=0.007), higher median white blood cell (WBC) counts at presentation (25 versus 8, P=0.0002), and higher peripheral median percentage of blast counts (63 versus 40, P=0.023). FAB subtype L2 (70% versus 49%, P=0.001) and CALLA-positive pre-B immunophenotype (75% versus 37%, P<0.001) predominated among Ph-positive ALL. Myeloid marker coexpression was more frequent in Ph-positive ALL when compared with Ph-negative ALL (52% vs. 27% for CD13, P<0.001, and 44% vs. 27% for CD33, P=0.005). Among patients treated with hyper-CVAD, the complete remission (CR) rate was 90% versus 55% (P=0.002) with pre-hyper-CVAD regimens (VAD and AML-like induction protocols), the median CR duration was 43 weeks versus 32 weeks (P>0.5), median disease-free survival (DFS) was 42 weeks versus 29 weeks (P=0.008), and median survival was 66 weeks versus 45 weeks (P>0.5). Patients with hyperdiploid Ph-positive ALL on hyper-CVAD therapy achieved significantly longer CR duration and DFS than hypo- and pseudodiploid cases (59 weeks versus 42 and 31 weeks, P=0.02 and 0.04, respectively). In contrast, patients treated with regimens prior to hyper-CVAD had significantly shorter CR duration (21 weeks versus 33 and 29 weeks, P=0.03) and DFS with hyperdiploid karyotypes when compared to pseudodiploid and hypodiploid cases (16 weeks versus 30 and 13 weeks, P=0.008). In conclusion, our results demonstrate improved response rate and DFS with current intensive regimens (hyper-CVAD) in patients with Ph-positive ALL, but no advantage in overall survival. <50> UI - 20138967 AU - Massa MJ AU - Iniesta P AU - Gonzalez-Quevedo R AU - de Juan C AU - Caldes T AU - Sanchez-Pernaute A AU - Cerdan J AU - Torres AJ AU - Balibrea JL AU - Benito M IN - Department of Biochemistry and Molecular Biology, Faculty of Pharmacy, Complutense University, Madrid, Spain. TI - Differential prognosis of replication error phenotype and loss of heterozygosity in sporadic colorectal cancer. SO - European Journal of Cancer 1999 Nov;35(12):1676-82 AB - Several distinct genetic alterations have been associated with colorectal tumorigenesis. This study investigated the frequency of microsatellite instability, also known as replication error (RER), and loss of heterozygosity (LOH) at six chromosome regions in sporadic colorectal cancer (CRC). Eighty-six tumour and paired normal mucosa samples were included in the study. A polymerase chain reaction (PCR)-based technique was performed to analyse six (CA)n dinucleotide repeats located near or within regions containing important genes implicated in the complex process of colorectal tumorigenesis (chromosomes 2p, 3p, 5q, 11p, 17p and 18q). Overall, LOH frequency was higher in RER-tumours (25/46, 54.3%) compared with RER+ tumours (9/40, 22.5) (P = 0.04). To investigate prognostic implications, survival analysis was performed for 66 patients. Compared with RER- tumours, patients with RER+ tumours at 2p, 3p, 5q, 11p or 18q were found to have an improved prognosis (overall survival, P = 0.02 and disease-free survival (DFS) P = 0.005) this variable being an independent prognostic factor by multivariate analysis (P = 0.001). Overall survival of patients whose tumours were LOH+ was significantly shorter compared with those without LOH (overall survival, P = 0.008 and DFS, P = 0.01). Thus, tumours displaying RER+ and LOH+ phenotype, as established by microsatellite analysis, show a differential prognosis. These data indicate that this may be a useful tool for the identification of patients at different risks affected by CRC. <51> UI - 20128886 AU - Paterakos M AU - Watkin WG AU - Edgerton SM AU - Moore DH 2nd AU - Thor AD IN - Department of Pathology, Evanston Northwestern Healthcare and Northwestern University Medical School, IL 60201, USA. TI - Invasive micropapillary carcinoma of the breast: a prognostic study. SO - Human Pathology 1999 Dec;30(12):1459-63 AB - Invasive micropapillary carcinoma (IMC) of the breast is a rare variant of infiltrating ductal carcinoma that has been associated with an extremely high incidence of lymph node metastases. Follow-up studies on patients with pure IMC breast cancer histology have been limited by low patient numbers, short duration of follow-up, and a lack of multivariate analyses. Using invasive breast cancers from 1,287 patients (median follow-up, 13.8 years), histological review showed 21 cases (1.7%) with pure IMC histology. Pure IMC histology was associated with high-grade histology (P = .04), metastases to regional lymph nodes (P < .001), a high mitotic index (P = .02), and erbB-2 immunopositivity (P = .007). Univariate analyses showed a strong association between IMC histology and shortened survival (disease-free survival [DFS], P = .0052; median, 44 months for IMC and 63 months for non-IMC; disease-specific survival [DSS], P = .014; medians, 71 and 78 for IMC and non-IMC, respectively) only in an analysis of all patients. Because only 1 case of node-negative IMC histology was available, univariate analysis of IMC histology was performed only on node-positive patients without significance. Multivariate analyses comparing IMC histology with either node-positive or all other breast cancers failed to show independent prognostic significance. In summary, breast cancer patients with pure IMC histology showed survival rates similar to those of other patients with equivalent numbers of lymph node metastases. <52> UI - 20118171 AU - Tobin WR AU - Greene RS IN - Department of Biological Sciences, State University of New York at Buffalo 14260, USA. wtobin@rocketmail.com TI - Meso-substituted cationic porphyrins interact with dsDNA and exhibit different localization patterns in radiation-induced fibrosarcoma cells. SO - Anticancer Research 1999 Jul-Aug;19(4B):2953-8 AB - Meso-substituted cationic porphyrins were examined for binding dsDNA. Subcellular localization time studies used Confocal Laser Scanning Microscopy of radiation-induced fibrosarcoma (RIF) cells incubated with porphyrins. Binding studies revealed a reversible interaction between porphyrin and dsDNA that is a function of DNA shape. Binding was inhibited at high salt concentrations, and enhanced by heat and DNA denaturants such as dimethyl formamide (DMF). Trans dicationic porphyrin required more stringent binding conditions than cis dicationic and tetracationic porphyrins. Phenol extraction of porphyrin from the DNA-porphyrin complex demonstrates that cationic porphyrins do not damage dsDNA at high concentrations. Localization studies within a 24-hour range reveal different distribution patterns. Metal chelates of tetracationic porphyrin exhibited a cytoplasmic localization with the exception of the zinc chelate. Localization of other metal chelates appears to be redistributed to lysosomes and mitochondria between 3 and 6 hours post-incubation. HPPH used in PDT clinical trials localizes to the cytoplasmic compartment. <53> UI - 20119001 AU - Munday R AU - Smith BL AU - Munday CM IN - AgResearch, Ruakura Agricultural Research Centre, Hamilton, New Zealand. mundayr@agresearch.cri.nz TI - Effect of inducers of DT-diaphorase on the toxicity of 2-methyl- and 2-hydroxy-1,4-naphthoquinone to rats. SO - Chemico-Biological Interactions 1999 Dec 15;123(3):219-37 AB - It has previously been shown that rats pre-treated with butylated hydroxyanisole (BHA), a well-known inducer of the enzyme DT-diaphorase, are protected against the toxic effects of 2-methyl-1,4-naphthoquinone but are made more susceptible to the harmful action of 2-hydroxy-1,4-naphthoquinone. In the present experiments, the effects of BHA have been compared with those of other inducers of DT-diaphorase. Rats were dosed with BHA, butylated hydroxytoluene (BHT), ethoxyquin (EQ), dimethyl fumarate (DMF) or disulfiram (DIS) and then challenged with a toxic dose of the naphthoquinones. All the inducers protected against the haemolytic anaemia induced by 2-methyl-1,4-naphthoquinone in rats, with BHA, BHT and EQ being somewhat more effective than DMF and DIS. A similar order of activity was recorded in the relative ability of these substances to increase hepatic activities of DT-diaphorase, consistent with a role for this enzyme in facilitating conjugation and excretion of this naphthoquinone. In contrast, all the compounds increased the haemolytic activity of 2-hydroxy-1,4-naphthoquinone. DMF and DIS were significantly more effective in this regard than BHA, BHT and EQ. DMF and DIS also caused a much greater increase in levels of DT-diaphorase in the intestine, suggesting that 2-hydroxy-1,4-naphthoquinone is activated by this enzyme in the gut. BHA, BHT and EQ had no effect on the nephrotoxicity of 2-hydroxy-1,4-naphthoquinone, but the severity of the renal lesions was decreased in rats pre-treated with DMF and DIS. The results of the present experiments show that modulation of tissue levels of DT-diaphorase may not only alter the severity of naphthoquinone toxicity in vivo, but may also change the relative toxicity of these substances to different target organs. <54> UI - 20090585 AU - Weiss IM AU - Kaufmann S AU - Mann K AU - Fritz M IN - Physik Department der TU-Munchen, Institut fur Biophysik, E22, James-Franck-Strasse, Garching, 85747, Germany. TI - Purification and characterization of perlucin and perlustrin, two new proteins from the shell of the mollusc Haliotis laevigata. SO - Biochemical & Biophysical Research Communications 2000 Jan 7;267(1):17-21 AB - Two new proteins, named perlucin and perlustrin, with M(r) 17,000 and 13,000, respectively, were isolated from the shell of the mollusc Halotis laevigata (abalone) by ion-exchange chromatography and reversed-phase HPLC after demineralization of the shell in 10% acetic acid. The sequence of the first 32 amino acids of perlucin indicated that this protein belonged to a heterogeneous group of proteins consisting of a single C-type lectin domain. Perlucin increased the precipitation of CaCO(3) from a saturated solution, indicating that it may promote the nucleation and/or the growth of CaCO(3) crystals. With pancreatic stone protein (lithostathine) and the eggshell protein ovocleidin 17, this is the third C-type lectin domain protein isolated from CaCO(3) biominerals. This indicates that this type of protein performs an important but at present unrecognized function in biomineralization. Perlustrin was a minor component of the protein mixture and the sequence of the first 33 amino acids indicated a certain similarity to part of the much larger nacre protein lustrin A. Copyright 2000 Academic Press. <55> UI - 20088903 AU - Ingram DA AU - Yang FC AU - Travers JB AU - Wenning MJ AU - Hiatt K AU - New S AU - Hood A AU - Shannon K AU - Williams DA AU - Clapp DW IN - Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana 46202, USA. TI - Genetic and biochemical evidence that haploinsufficiency of the Nf1 tumor suppressor gene modulates melanocyte and mast cell fates in vivo. SO - Journal of Experimental Medicine 2000 Jan 3;191(1):181-8 AB - Neurofibromatosis type 1 (NF1) is a common autosomal-dominant disorder characterized by cutaneous neurofibromas infiltrated with large numbers of mast cells, melanocyte hyperplasia, and a predisposition to develop malignant neoplasms. NF1 encodes a GTPase activating protein (GAP) for Ras. Consistent with Knudson's "two hit" model of tumor suppressor genes, leukemias and malignant solid tumors in NF1 patients frequently demonstrate somatic loss of the normal NF1 allele. However, the phenotypic and biochemical consequences of heterozygous inactivation of Nf1 are largely unknown. Recently neurofibromin, the protein encoded by NF1, was shown to negatively regulate Ras activity in Nf1-/- murine myeloid hematopoietic cells in vitro through the c-kit receptor tyrosine kinase (dominant white spotting, W). Since the W and Nf1 locus appear to function along a common developmental pathway, we generated mice with mutations at both loci to examine potential interactions in vivo. Here, we show that haploinsufficiency at Nf1 perturbs cell fates in mast cells in vivo, and partially rescues coat color and mast cell defects in W(41) mice. Haploinsufficiency at Nf1 also increased mast cell proliferation, survival, and colony formation in response to Steel factor, the ligand for c-kit. Furthermore, haploinsufficiency was associated with enhanced Ras-mitogen-activated protein kinase activity, a major downstream effector of Ras, via wild-type and mutant (W(41)) c-kit receptors. These observations identify a novel interaction between c-kit and neurofibromin in vivo, and offer experimental evidence that haploinsufficiency of Nf1 alters both cellular and biochemical phenotypes in two cell lineages that are affected in individuals with NF1. Collectively, these data support the emerging concept that heterozygous inactivation of tumor suppressor genes may have profound biological effects in multiple cell types. <56> UI - 20107053 AU - Hillman JD AU - Brooks TA AU - Michalek SM AU - Harmon CC AU - Snoep JL AU - van Der Weijden CC IN - Department of Oral Biology, University of Florida College of Dentistry, Gainesville, Florida 32610, USA. jhillman@dental.ufl.edu TI - Construction and characterization of an effector strain of Streptococcus mutans for replacement therapy of dental caries. SO - Infection & Immunity 2000 Feb;68(2):543-9 AB - An effector strain has been constructed for use in the replacement therapy of dental caries. Recombinant DNA methods were used to make the Streptococcus mutans supercolonizing strain, JH1140, lactate dehydrogenase deficient by deleting virtually all of the ldh open reading frame (ORF). To compensate for the resulting metabolic imbalance, a supplemental alcohol dehydrogenase activity was introduced by substituting the adhB ORF from Zymomonas mobilis in place of the deleted ldh ORF. The resulting clone, BCS3-L1, was found to produce no detectable lactic acid during growth on a variety of carbon sources, and it produced significantly less total acid due to its increased production of ethanol and acetoin. BCS3-L1 was significantly less cariogenic than JH1140 in both gnotobiotic- and conventional-rodent models. It colonized the teeth of conventional rats as well as JH1140 in both aggressive-displacement and preemptive-colonization models. No gross or microscopic abnormalities of major organs were associated with oral colonization of rats with BCS3-L1 for 6 months. Acid-producing revertants of BCS3-L1 were not observed in samples taken from infected animals (reversion frequency, <10(-3)) or by screening cultures grown in vitro, where no revertants were observed among 10(5) colonies examined on pH indicator medium. The reduced pathogenic potential of BCS3-L1, its strong colonization potential, and its genetic stability suggest that this strain is well suited to serve as an effector strain in the replacement therapy of dental caries in humans. <57> UI - 20081019 AU - Ogawa T AU - Dobrinski I AU - Avarbock MR AU - Brinster RL IN - Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, 3850 Baltimore Ave., Philadelphia, Pennsylvania 19104, USA. TI - Transplantation of male germ line stem cells restores fertility in infertile mice [see comments]. CM - Comment in: Nat Med 2000 Jan;6(1):16-7 SO - Nature Medicine 2000 Jan;6(1):29-34 AB - Azoospermia or oligozoospermia due to disruption of spermatogenesis are common causes of human male infertility. We used the technique of spermatogonial transplantation in two infertile mouse strains, Steel (Sl) and dominant white spotting (W), to determine if stem cells from an infertile male were capable of generating spermatogenesis. Transplantation of germ cells from infertile Sl/Sld mutant male mice to infertile W/Wv or Wv/W54 mutant male mice restored fertility to the recipient mice. Thus, transplantation of spermatogonial stem cells from an infertile donor to a permissive testicular environment can restore fertility and result in progeny with the genetic makeup of the infertile donor male. <58> UI - 20105413 AU - Yokozawa T AU - Towatari M AU - Iida H AU - Takeyama K AU - Tanimoto M AU - Kiyoi H AU - Motoji T AU - Asou N AU - Saito K AU - Takeuchi M AU - Kobayashi Y AU - Miyawaki S AU - Kodera Y AU - Ohno R AU - Saito H AU - Naoe T IN - First Department of Internal Medicine, Nagoya University School of Medicine, Nagoya, Japan. TI - Prognostic significance of the cell cycle inhibitor p27Kip1 in acute myeloid leukemia. SO - Leukemia 2000 Jan;14(1):28-33 AB - There are few molecular biologic determinants that are prognostic for patients with acute myeloid leukemia (AML). Hence, we examined whether cellular levels of the cyclin-dependent kinase inhibitor p27Kip1 in acute myeloid leukemia could be used to predict clinical outcome in AML. Using immunoblot analysis, levels of p27 were assessed in blast cells from 72 AML patients who were registered and treated by the identical chemotherapy protocol. AML cases were classified into three groups on the basis of the percentage of the expression level of p27 compared to a control cell line. AML cases exhibiting p27 expression at low, moderate, and high levels were 43, 9, and 20 cases, respectively. No significant differences in the rates of complete remission (CR) were observed among the three groups. Although the level of p27 expression was not correlated with any other possible prognostic markers, such as age, white blood cell count, chromosome abnormalities, and FAB subclasses, patients with high p27 expression had a significantly increased disease-free survival (DFS) (78% vs 19%, P = 0.004). We further examined the expression of cyclin E at the protein level in all 72 AML cases. We observed a statistically significant correlation between a high cyclin E level and a high p27 level (P < 0.005). However, we failed to find any correlation between the rates of CR or DFS and cyclin E expression. The present study reveals that levels of p27 expression can be one of the useful prognostic molecular markers for AML. Leukemia (2000) 14, 28-33. <59> UI - 20067241 AU - Slavkin HC IN - National Institute of Dental and Craniofacial Research, Bethesda, Md. 20892-2290, USA. TI - Streptococcus mutans, early childhood caries and new opportunities. SO - Journal of the American Dental Association 1999 Dec;130(12):1787-92 <60> UI - 20080215 AU - Lamoreux ML IN - Department of Veterinary Pathobiology, Texas A&M University, College Station 77843, USA. llamoreux@cvm.tamu.edu TI - Strain-specific white-spotting patterns in laboratory mice. SO - Pigment Cell Research 1999 Dec;12(6):383-90 AB - White spotting is the absence of melanocytes (pigment cells) from part or all of the locations in the body where they are normally found. At least in the case of the W (kit) locus, white spotting has been attributed to apoptosis. In addition to the death of melanoblasts, white spotting might result from their failure to migrate to their normal locations. These developmental failures are known to be melanocyte-specific in some instances and environment-specific in others. The environment is defined as the tissues surrounding the melanoblast. Patterns of white spotting were examined on mice mutant at the piebald (s), patch (Ph), dominant spotting (W(J2)) rumpwhite (Rw) or belted (bt) lo