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Zebrafish (Danio rerio) is used as a model system for in vivo studies. To expand the research scope of physical, biochemical and physiological studies, a cold-tolerant model of zebrafish was developed. The common carp (Cyprinus carpio) muscle form of creatine kinase (CK, EC 2.7.3.2) can maintain enzymatic activity at a temperature of around 15°C. However, a cold-inducible promoter of zebrafish, hsc 70 (heat shock protein 70 cognate), is able to increase the expression of gene product by 9.8 fold at a temperature of 16°C. Therefore, the carp CK gene was promoted by hsc 70 and transfected into zebrafish embryos. Resulting transgenic zebrafish survived and could maintain its swimming behavior at 13°C, which was not possible with the wild-type zebrafish. The swimming distance of the transgenic fish was 42% greater than that of the wild type at 13°C. This new transgenic fish model is ideal for studies of ectothermal vertebrates in low-temperature environments.

The endocervix in the female reproductive tract (FRT) is susceptible to sexually transmitted pathogens such as Chlamydia trachomatis and Neisseria gonorrhoeae. Endocervical epithelial cells in vivo make innate immune mediators that likely aid in the protection from these pathogens. In vitro studies to investigate the innate epithelial cell immune response to endocervical pathogens have been hindered by the paucity of human endocervix-derived epithelial cell lines that display the differentiation proteins and functional characteristics of their site of origin. We have established an immortalized epithelial cell line (A2EN) derived from an endocervical tissue explant that can be polarized to exhibit distinct apical and basolateral membrane domains. Polarized A2EN cells secrete mucus at their apical surface, and express MUC5B, a mucin specific to the endocervix. Polarized A2EN cells also express hormone receptors that respond appropriately to female steroid hormones. Polarized A2EN cells can be stimulated with the toll-like receptor 3 agonist, polyI:C, to express anti-microbial peptides (AMPs) as well as pro-inflammatory cytokines and chemokines. Cytokines and chemokines are also differentially secreted depending on the hormone milieu in which the cells are exposed. We conclude that polarized A2EN cells maintain distinctive phenotypic and functional characteristics of the epithelial cells found in the endocervix and, hence, could provide a useful, new in vitro model system for investigations on the role of endogenous and exogenous factors that regulate endocervical epithelial cell immunity including studies on sexually transmitted infections and topical microbicides., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

Background: Concurrent sexually transmitted infections (STIs) increase the likelihood of HIV transmission. The levels of defensins are frequently elevated in genital fluids from individuals with STIs. We have previously shown that human defensins 5 and 6 (HD5 and HD6) promote HIV entry and contribute to Neisseria gonorrhoeae-mediated enhancement of HIV infectivity in vitro. In this study, we dissect the molecular mechanism of the HIV enhancing effect of defensins., Results: HD5 and HD6 primarily acted on the virion to promote HIV infection. Both HD5 and HD6 antagonized the anti-HIV activities of inhibitors of HIV entry (TAK 779) and fusion (T-20) when the inhibitors were present only during viral attachment; however, when these inhibitors were added back during viral infection they overrode the HIV enhancing effect of defensins. HD5 and HD6 enhanced HIV infectivity by promoting HIV attachment to target cells. Studies using fluorescent HIV containing Vpr-GFP indicated that these defensins enhanced HIV attachment by concentrating virus particles on the target cells. HD5 and HD6 blocked anti-HIV activities of soluble glycosaminoglycans including heparin, chondroitin sulfate, and dextran sulfate. However, heparin, at a high concentration, diminished the HIV enhancing effect of HD5, but not HD6. Additionally, the degree of the HIV enhancing effect of HD5, but not HD6, was increased in heparinase-treated cells. These results suggest that HD5 and haparin/heparan sulfate compete for binding to HIV., Conclusions: HD5 and HD6 increased HIV infectivity by concentrating virus on the target cells. These defensins may have a negative effect on the efficacy of microbicides, especially in the setting of STIs.

Background & Aims: RUNX3 is a tumor suppressor originally identified in gastric cancer. The mutation R122C in RUNX3 promotes gastric carcinogenesis by unclear mechanisms. We investigated how Runx3-deficiency contributes to distinct changes in the gastric epithelium that precede neoplasia., Methods: Runx3-deficient (Runx3(-/-)) and wild-type BALB/c adult mice were subjected to histological analyses. Gastric cancer formation after administration of N-methyl-N-nitrosourea was evaluated. Runx3(+/+) and Runx3(-/-) gastric epithelial cell lines were used to investigate the molecular basis underlying Runx3 function., Results: The gastric epithelia in Runx3(-)/(-) adult mice was hyperplastic, with loss of chief cells and development of mucin 6- and trefoil factor-2-expressing metaplasia. The gastric epithelium of Runx3(-)/(-) mice had an intestinal phenotype that expressed Cdx2. After addition of N-methyl-N-nitrosourea, Runx3- mice, unlike wild-type mice, consistently developed adenocarcinomas, indicating that Runx3-deficiency leads to premalignant changes in the gastric epithelia. RUNX3, but not the RUNX3 mutant R122C, repressed Cdx2 expression by attenuation of oncogenic beta(symbol)-catenin and Tcfs., Conclusions: Runx3-deficiency leads to a precancerous state in the gastric epithelia of mice, characterized by loss of chief cells but not parietal cells; inflammation did not appear to be involved., (Copyright © 2011 AGA Institute. Published by Elsevier Inc. All rights reserved.)

Recombinant human bone morphogenetic protein-2 (rhBMP2) has been shown to induce both in vitro osteogenic differentiation and in vivo bone formation, with the capacity of rhBMP2 to elicit the repair of numerous bony defects (calvaria, spinal fusion, femora, and so on) well documented. In addition, rhBMP2 has been approved by the Food and Drug Administration (FDA) for selected human indications. Despite the fact that healing is often achieved, the challenge still remains to optimize the therapeutic use of rhBMP2. One avenue may be through the combination of rhBMP2 with stem cells capable of osteogenic differentiation. This study investigates the ability of rhBMP2 at various doses in combination with human adipose-derived stem cells (ASCs) to heal critical-sized rat segmental femoral defects. For this, different doses of rhBMP2 were incorporated with apatite-coated porous poly(l-lactide-co-dl-lactide) (70  :  30) (PLDLA) scaffolds, seeded with ASCs, and implanted into athymic rats. After 8 weeks, all implants were harvested and processed for bone formation using micro computed tomography (microCT) analysis and histology. Despite the findings that indicate no adverse effect of the apatite surface on ASC osteogenesis, no significant difference in bone formation could be qualitatively or quantitatively determined upon the implantation of ASC-seeded scaffolds absorbed to increasing doses of rhBMP2. Such results would suggest that the presence of ASCs within rhBMP2-absorbed scaffolds does not improve the bone-forming ability of the construct and that the formation of bone may be driven by the rhBMP2 alone. Based on these results, the addition of ASCs to rhBMP2-treated scaffolds may provide no significant advantage in terms of the ability to heal bone.

Objective: To determine the impact of dental prosthetic condition on food consumption, risk of malnutrition and follow-up 4-year mortality risk in elderly Taiwanese., Design and Setting: Analyzing the data sets of the 1999 and 2003 "Survey of Health and Living Status of the Elderly in Taiwan", a longitudinal cohort study., Participants: A national probability sample of 2766 men and women 65 years of age or older., Measurements: Self-reported intake frequencies of major food categories, masticatory ability, food consumption, and the risk of malnutrition assessed with the Mini Nutritional Assessment (short-form) stratified by dental prosthetic condition. Cox regression was used to compare follow-up mortality risk., Results: Non-denture wearers and removable-denture wearers had poorer masticatory ability and greater nutritional risk and consumed fruits and vegetable less often compared to fixed-denture wearers. Removable-denture wearers also had lower self-perceived nutritional status and BMI compared to fixed-denture wearers. Survival analysis showed that non-denture wearers and removable-denture wearers had lower follow-up 4-year survival. Cox regression analysis showed that removable-denture wearers had increased follow-up 4-year mortality risk compared to fixed-denture wearers adjusted for sociodemographic, lifestyle and health-related factors., Conclusions: Based on data of a national sample of a longitudinal cohort study, dental prosthetic condition is a significant factor of nutritional health in the elderly. It can affect food pattern and the risk of malnutrition and mortality of elderly persons. Dental care should be an important part of geriatric health promotion program and fixed-denture is a preferred choice over removable-denture.

Objective: The study was to evaluate the ability of the Mini Nutritional Assessment in predicting malnutrition in people with three subtypes of mental disorder (schizophrenia, major depression and bipolar disorder) in Taiwan., Design: The study involved a convenience sample of 120 residents of psychiatric wards managed by a hospital in central Taiwan (52 with schizophrenia, 36 with major depression and 32 with bipolar disorder) classified according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria., Methods: A structured questionnaire elicited subjects' personal data, disease history and answers to questions in the Mini Nutritional Assessment. Serum and anthropometrical parameters were measured. Nutritional status was evaluated with a content-equivalent version of the Mini Nutritional Assessment (Taiwan version-1, T1)., Results: The Mini Nutritional Assessment-Taiwan version-1 was effective in assessing the nutritional status of people of all three subtypes of disorder. Nutritional statuses predicted with the Mini Nutritional Assessment-Taiwan version-1 agreed well with other nutritional indicators such as BMI, waist circumference and appetite status. According to the Mini Nutritional Assessment-Taiwan version-1, people with major depression were more likely to be at risk of undernutrition, whereas people with schizophrenia or bipolar disorder were more likely to be at risk of overnutrition., Conclusion: The Mini Nutritional Assessment-Taiwan version-1 can effectively grade both undernutrition and overnutrition of people with schizophrenia, major depression or bipolar disorder., Relevance to Clinical Practice: The Mini Nutritional Assessment enables nurses to monitor emerging nutritional problems in people with psychiatric disorder without relying on subjective judgement. With proper intervention, it can help reduce nutrition-related chronic conditions in these individuals and save on healthcare cost., (© 2011 Blackwell Publishing Ltd.)

BMI, mid-arm circumference (MAC) and calf circumference (CC) are anthropometric indicators often included in geriatric health measurement scales. However, their relative effectiveness in predicting long-term mortality risk has not been extensively examined. The present study aimed to evaluate the relative effectiveness of these anthropometrics in predicting long-term mortality risk in older adults. The study prospectively analysed the ability of these indicators in predicting 4-year follow-up mortality risk of a population-representative sample of 4191 men and women, 53 years of age or older in the 'Survey of Health and Living Status of the Elderly in Taiwan'. Cox regression analyses were performed to evaluate the association of follow-up mortality risk with low ( < 21 kg/m2) or high ( ≥ 27 kg/m2) BMI, low MAC ( < 23·5/22 cm for men/women) and low CC ( < 30/27 cm) respectively, according to Taiwanese-specific cut-off points. Results showed that low CC and low MAC were more effective than low BMI in predicting follow-up mortality risk in 65-74-year-old elderly. But low CC and low BMI were more effective than low MAC in ≥ 75-year-old elderly, and low BMI was more effective than low MAC or low CC in 53-64-year-old persons. High BMI was not effective in predicting mortality risk in any of these age ranges. These results suggest that in elderly adults, CC is more effective than BMI in predicting long-term mortality risk. Thus, more consideration to CC and MAC in designing geriatric health or nutritional measurement scales is recommended.

Liver cancer patients are confronted with the additional risk of malnutrition because the disease is often associated with hepatitis, liver cirrhosis, and metabolic disturbances. Nutritional intervention can improve treatment outcome, but early detection is important. This study aimed to determine whether the Mini Nutritional Assessment (MNA) could effectively rate the nutritional status of patients with liver cancer in Taiwan. A total of 300 patients were evaluated for nutritional status with two modified versions of the MNA in short and long forms. MNA-Taiwan Version 1 adopted population-specific anthropometric cutpoints, whereas Version 2 replaced mid-arm and calf circumferences in place of body mass index. Predicted statuses were compared to results predicted by the Council on Nutrition Appetite Questionnaire (CNAQ) and analyzed for correlations with biochemical or cancer status parameters. Results showed that both versions of the MNA were effective in predicting nutritional status, and predictions by the short forms agreed well with those by the long forms. The nutritional scores correlated well with hemoglobin, serum albumin, C-reactive protein, r-glutamyl transpeptidase, TNM (tumor, node, metastasis) staging, and severity of cirrhosis. These results suggest that the MNA can be an effective tool for assessing the nutritional status of patients with liver cancer.

Defensins are highly abundant antimicrobial peptides in the female genital mucosa. We have previously shown that human defensins 5 and 6 (HD5 and HD6), produced by cervicovaginal epithelial cells, significantly enhance HIV infectivity in vitro. Candidate polyanion microbicides, including PRO 2000, cellulose sulfate and carrageenan, failed to protect women against HIV infection in large-scale clinical trials, but the molecular basis of ineffectiveness was not clear. We hypothesized that mucosal host factors such as HD5 an HD6 may alter the activity of polyanion microbicides against HIV. Our results demonstrated that HD5 and HD6 but not their linear analogs antagonized the anti-HIV activity of PRO 2000, cellulose sulfate and carrageenan in vitro. Polyanion microbicides also reduced the HIV-enhancing effect of these defensins. We conclude that mucosal host factors could negatively impact the efficacy of topical microbicides against HIV, and their impact on the activity of candidate microbicides needs to be considered during the preclinical evaluation., (Copyright © 2010 S. Karger AG, Basel.)

Study Design: Retrospective review., Objective: Anchor stability and prominence are problems with pelvic fixation in pediatric spinal deformity surgery. We compared the new sacral alar iliac (SAI) fixation technique (with a starting point in the sacral ala and in-line anchors deep under the midline muscle flap) with other methods of screw fixation., Summary of Background Data: Iliac anchors have been shown to provide the best form of pelvic fixation. A trajectory from the posterior sacral surface to the iliac wings has recently been described. To our knowledge, no clinical series has compared this method of pelvic fixation in children to others., Methods: Of 32 consecutive pediatric patients who underwent SAI fixation, 2 died and 26 returned for follow-up (>2 years). Mean age at surgery was 14 years. Average screw size was 67 mm long and 7 to 9 mm in diameter. Clinical examinations, radiographs, and computed tomography scans were analyzed. Outcomes included pain over the implants, screw placement, implant prominence, radiographic lucency, need for revision, and infection. SAI patients were compared with 27 previous patients who had pelvic fixation via other screw techniques., Results: For SAI fixation, correction of pelvic obliquity and Cobb angles were 20° ± 11° (70%) and 42° ± 25° (67%), respectively. For other pelvic fixation methods, those values were 10° ± 9° (50%), and 46° ± 16° (60%), respectively. Compared with other screws, SAI screws provided significantly better pelvic obliquity correction (P = 0.002) but no difference in Cobb correction. There were 2 lucencies adjacent to screws in both groups. Computed tomography scans of 18 SAI patients showed no intrapelvic protrusion, but 1 screw extended laterally (<5 mm). One early SAI patient required revision with larger screws, which relieved pain; there was 1 revision in the comparison group. SAI patients had no deep infections, implant prominence, late skin breakdown, or anchor migration; traditional patients had 3 deep infections (P = 0.09) and 3 instances of implant prominence, skin breakdown, or anchor migration., Conclusion: SAI pelvic fixation produces better correction of pelvic obliquity than do previous techniques. Radiographic and clinical anchor stability is satisfactory at 2-year follow-up.

A simple, reliable, noninvasive, and easy-to-use instrument is important for successful monitoring of emerging nutrition problems in elderly people. The objectives of this study were to determine whether adoption of population-specific body mass index (BMI) cutpoints would improve the predictive ability of the short-form Mini Nutritional Assessment (MNA) and whether calf circumference could be an acceptable alternative to BMI in the short-form MNA without compromising its predictive ability. Using convenience sampling, the study recruited 301 community-living, 109 care center-living, and 68 nursing home-living elderly people, 65 years or older, as subjects. Subjects were evaluated with the short-form MNA in three versions: (a) the original, (b) Taiwan version 1 (T1), that adopted population-specific BMI cutpoints, and (c) Taiwan version 2 (T2), which substituted calf circumference for BMI, and with the long-form MNA-T2 as a reference. The ability of the short forms to predict the long-form MNA-T2 was evaluated with binary classification and analysis of receiver operating characteristic curves. Results were analyzed with an SPSS for Windows 12.0 software package (version 12.0.1C, 2000, SPSS Inc, Chicago, IL). Results showed that adoption of population-specific anthropometric BMI cutpoints improved the predictive ability of the short-form MNA, whereas replacement of BMI with calf circumference further improved the predictive ability of the scale (kappa values of the binary classification tests were 0.596, 0.742, and 0.843 for community-living; 0.560, 0.683, and 0.839 for care center-living; and 0.346, 0.454, and 0.522 for nursing home-living elderly for the original, T1, and T2 MNA short-form versions, respectively). These results suggest that modification of a measurement tool according to cultural or anthropometric features of the target population is necessary. The study also shows that calf circumference can be an acceptable alternative to BMI in the short-form MNA. Because measuring calf circumference is easier and less time-consuming then measuring BMI, the modified tool can make periodic nutritional screening an easier job and routine nutritional screening a more realistic goal in geriatric care., (Copyright 2010 American Dietetic Association. Published by Elsevier Inc. All rights reserved.)

Proteasome inhibition is one of the new pharmaceutical approaches to study antitumor activity. Although the active components are not yet identified for either treating or preventing cancer, the low toxicity plant Anisomeles ovata R. Br. has been used in traditional herbal medicine for more than 1000 years. In this study, the methanol extract from Anisomeles ovata showed potent inhibition of proteasome activity. Twenty compounds, two macrocycylic diterpenoids, six aromatics, seven flavonoids, and five phenylethanoids were isolated from Fang Feng Cao the dried aerial parts of A. ovata. Their structures have been established on the basis of spectral evidence. Using a proteolysis assay for inhibition of 26S proteasome from pig red blood cells, we found that 5,6,3',4'-tetrahydroxy-7-methoxyflavone inhibited 90.5 %, 85.4 % and 73.1 % the chymotrypsin-like, caspase-like and trypsin-like activities of 26S proteasome with IC (50) values of 14.0, 5.4 and 24.1 microM, respectively, when Suc-LLVY-AMC, Z-LLE-AMC and Ac-RLR-AMC were used as substrates. 5,6,3',4'-Tetrahydroxy-7-methoxyflavone had a higher inhibitory at 15 minutes. A combination of 5,6,3',4'-tetrahydroxy-7-methoxyflavone and 5,6,4'-trihydroxy-7,3'-dimethoxyflavone increased the inhibition ability on 26S enzymatic activity. This combination appears to be a potentially attractive chemotherapy approach. We have found that 5,6,3',4'-tetrahydroxy-7-methoxyflavone has the highest inhibitory effects on 26S proteasome activities when compared to the other 11 flavonoids. These results suggest that both the 6-hydroxy and 7-methoxy positions of the flavone may play an important role in targeting 26S proteasome activity., (Georg Thieme Verlag KG Stuttgart.New York.)

Sexually transmitted infections increase the likelihood of HIV-1 transmission. We investigated the effect of Neisseria gonorrheae (gonococcus [GC]) exposure on HIV replication in primary resting CD4(+) T cells, a major HIV target cell during the early stage of sexual transmission of HIV. GC and TLR2 agonists, such as peptidylglycan (PGN), Pam(3)CSK(4), and Pam(3)C-Lip, a GC-derived synthetic lipopeptide, but not TLR4 agonists including LPS or GC lipooligosaccharide enhanced HIV-1 infection of primary resting CD4(+) T cells after viral entry. Pretreatment of CD4(+) cells with PGN also promoted HIV infection. Anti-TLR2 Abs abolished the HIV enhancing effect of GC and Pam(3)C-Lip, indicating that GC-mediated enhancement of HIV infection of resting CD4(+) T cells was through TLR2. IL-2 was required for TLR2-mediated HIV enhancement. PGN and GC induced cell surface expression of T cell activation markers and HIV coreceptors, CCR5 and CXCR4. The maximal postentry HIV enhancing effect was achieved when PGN was added immediately after viral exposure. Kinetic studies and analysis of HIV DNA products indicated that GC exposure and TLR2 activation enhanced HIV infection at the step of nuclear import. We conclude that GC enhanced HIV infection of primary resting CD4(+) T cells through TLR2 activation, which both increased the susceptibility of primary CD4(+) T cells to HIV infection as well as enhanced HIV-infected CD4(+) T cells at the early stage of HIV life cycle after entry. This study provides a molecular mechanism by which nonulcerative sexually transmitted infections mediate enhancement of HIV infection and has implication for HIV prevention and therapeutics.

Objective: The study was to determine whether a modified MNA (Mini Nutritional Assessment) which adopted population-specific anthropometric cut-points but without BMI could maintain its predicting ability in community-living elderly in Taiwan., Design: Purposive sampling., Setting: Community-living elderly., Participants: Three hundred and one (138 male and 163 female) > 65-year-old outpatients seeking free annual health examination at an area hospital in central Taiwan., Measurements: A structured questionnaire elicited personal data, lifestyle information and answers to the MNA. Laboratory results from health checkup provided the needed biochemical data. Each subject's nutritional status was assessed with the MNA in three versions: the original, the MNA-TI (with population-specific cut-points), and the MNA-TII (further eliminated the BMI question and redistributed its score to the MAC and CC questions)., Results: All three versions identified the same 0.7% elderly malnourished. The proportions predicted at risk of malnutrition were 16.6, 12.0 and 10% according to the original, MNA-TI and MNA-TII, respectively. Friedman Test and post hoc analysis indicated that the pattern predicted by the original MNA was different from that predicted by the two modified versions whereas the patterns predicted by the two modified versions were not different from each other., Conclusion: Adoption of population-specific anthropometric cut-points improves the predicting ability of the MNA in Taiwanese elderly, and the improved functionality is maintained in a version without BMI (but with adjusted MAC and CC scores). A MNA without BMI has greater applicability and can enhance professional efficiency of healthcare workers.

Macrophages are major HIV target cells. They support both productive and latent HIV-1 infection. Susceptibility of primary macrophages to HIV depends on the anatomical location and activation state of the cells. We demonstrate that peritoneal macrophages (PMs) are abundant in ascitic fluid of patients with liver cirrhosis and are susceptible to HIV-1 infection. PMs expressed CD68, a differentiation marker, exhibited phagocytic activity, and survived in culture for 2 months without additional growth factors. Freshly isolated PMs were susceptible to HIV-1 R5 strains but not to X4-T-cell line-adapted strains. Interestingly, after 7 days in culture, PMs acquired susceptibility to X4-T-cell line-adapted strains. HIV entry inhibitors, TAK779 and AMD3100, blocked HIV infection of PMs, indicating that infection by R5 and X4 strains was mediated by CCR5 and CXCR4, respectively. Although PMs did not express detectable cell surface levels of CXCR4 and CCR5, they did express mRNAs of these HIV coreceptors and responded to stimulation by their natural ligands, SDF-1alpha and RANTES. PMs were susceptible to HIV-1 X4, R5, and X4R5 primary isolates. PMs after 7 days in culture produced greater amounts of X4 and X4R5 HIV than freshly isolated PMs. The day-7 PMs were more susceptible to R5 infection in a single-cycle infection assay, but there was no increase in viral production in a multiple-round infection assay. The level of CXCR4 mRNA and production of CC-chemokines (MIP-1alpha, MIP-1beta, and RANTES) increased significantly during 7 days in culture. Our results indicate that PMs are susceptible to receptor-mediated infection by a broad range of HIV strains. These primary macrophages could provide a valuable system for investigating the role of primary macrophages in HIV pathogenesis.

Background & Aims: The transcription factor RUNX3 is a gastric tumor suppressor. Tumorigenic Runx3(-/-) gastric epithelial cells attach weakly to each other, compared with nontumorigenic Runx3(+/+) cells. We aimed to identify RUNX3 target genes that promote cell-cell contact to improve our understanding of RUNX3's role in suppressing gastric carcinogenesis., Methods: We compared gene expression profiles of Runx3(+/+) and Runx3(-/-) cells and observed down-regulation of genes associated with cell-cell adhesion in Runx3(-/-) cells. Reporter, mobility shift, and chromatin immunoprecipitation assays were used to examine the regulation of these genes by RUNX3. Tumorigenesis assays and immunohistological analyses of human gastric tumors were performed to confirm the role of the candidate genes in gastric tumor development., Results: Mobility shift and chromatin immunoprecipitation assays revealed that the promoter activity of the gene that encodes the tight junction protein claudin-1 was up-regulated via the binding of RUNX3 to the RUNX consensus sites. The tumorigenicity of gastric epithelial cells from Runx3(-/-) mice was significantly reduced by restoration of claudin-1 expression, whereas knockdown of claudin-1 increased the tumorigenicity of human gastric cancer cells. Concomitant expression of RUNX3 and claudin-1 was observed in human normal gastric epithelium and cancers., Conclusions: The tight junction protein claudin-1 has gastric tumor suppressive activity and is a direct transcriptional target of RUNX3. Claudin-1 is down-regulated during the epithelial-mesenchymal transition; RUNX3 might therefore act as a tumor suppressor to antagonize the epithelial-mesenchymal transition., (Copyright 2010 AGA Institute. Published by Elsevier Inc. All rights reserved.)

The ubiquitin-proteasome pathway is responsible for the degradation of most intracellular proteins in eukaryotes. It may also play a role in the modulation of inflammatory process and pathogenesis of cancer. Immunoglobulin levels are higher in cancer. Obesity is a risk factor for several common diseases, particularity type 2 diabetes mellitus, cardiovascular diseases, and tumors. The aim of this study was to study a possible correlation between plasma ubiquitin, 26S proteasome levels, and obesity. The body mass index (BMI), plasma ubiquitin levels, and 26S proteasome activity levels were determined and statistically analyzed in 31 volunteers, aged 19 to 58 years and including 9 men and 22 women, from the general population of Southern Taiwan. We also compared the immunoglobulin among the underweight, normal-weight, and overweight groups. We demonstrated that plasma ubiquitin is significantly decreased in obese individuals vs normal controls (65.2 +/- 23.4 vs 159.5 +/- 73.1 ng/mL). Plasma ubiquitin levels were found to be inversely correlated with the BMI of individuals (r = -0.39, P < .001). In addition, there was an inverse relationship between 20S proteasome levels in red blood cells and BMI (r = -0.33, P < .001), whereas 26S proteasome activity was found to be dependent quantitatively to S5a in erythrocytes (r = 0.88, P < .001). Immunoglobulin is significantly decreased in overweight individuals vs normal controls. Plasma ubiquitin and 20S proteasome levels are potential biomarkers for the risk assessment and possibly serve as one of the targeted studies for the development of human obesity.

Background: The Mini Nutritional Assessment (MNA) and the Malnutrition Universal Screening Tool (MUST) are two widely used malnutrition screening scales., Objective: The study was to compare the grading ability of the two scales, and to determine whether adoption of population-specific anthropometric cut-points could improve the grading ability of these tools and whether calf circumference (CC) could be an acceptable alternative to BMI in these scales., Design: Purposive sampling., Settings: Outpatients receiving annual physical examination at an area hospital., Participants: Community-living elderly who were 65 years or older, able to communicate orally, without acute health conditions and willing to sign a study-consent., Methods: Subjects were measured for anthropometrics and blood biochemical indicators, and interviewed for personal data and answers to the MNA and MUST. The risk of malnutrition was evaluated with the short-form MNA (MNA-SF) and the MUST, each in three versions (the original, Taiwan version-1 (T1) which adopted population-specific anthropometric cut-points, and Taiwan version-2 (T2) which replaced BMI with CC). Long-form (LF) MNA versions served as references., Results: Results showed that (a) in both scales, patterns of nutritional status rated with the original versions were different from those rated with respective modified versions but ratings made with two modified versions were the same, (b) the T2 versions showed the best grading ability based on agreement with the reference (MNA-LF), and (c) MNA-SF versions rated greater proportions of subjects at risk of malnutrition than the respective MUST versions., Conclusions: (a) Adoption of population-specific anthropometric cut-points improves the grading ability of the MNA-SF and the MUST in community-living Taiwanese, (b) CC is an acceptable alternative to BMI for both MNA-SF and MUST, and (c) nutritional assessment tools should be as much population or ethnically specific as possible to account for cultural and anthropometric differences across populations.

Inhibiting proteasomal degradation has been shown to induce apoptosis in tumor cells. Utilization of proteasome inhibition is therefore one approach to anticancer therapy. Some of the flavonoids can induce cell apoptosis via inhibiting proteasome 26S activity. In this study, the inhibition of 26S proteasome from pig red blood cells was analyzed on 12 flavones, 5 flavanones, and 9 isoflavones by using a proteolysis assay. Several flavonoids such as apigenin-6-hydroxy-7-O-beta-D-glucoside, quercetin, rutin, 6-hydroxyapigenin, 5,6,4'-trihydroxy-7,3'-dimethoxyflavone, 5,6,3',4'-tetrahydroxy-7-methoxyflavone, glycitecin, and 6,7,4'-trihydroxyisoflavone inhibited the chymotrypsin-like, caspase-like, or trypsin-like activity of 26S proteasome when Suc-LLVY-AMC, Z-LLE-AMC, and Ac-RLR-AMC were used as substrates. Three peptidase activities of flavonoids were found to be significantly correlated with one another. Flavones had significantly stronger inhibitory effects on chymotrypsin-like and caspase-like activities than flavanones and isoflavones. 5,6,3',4'-Tetrahydroxy-7-methoxyflavone, 5,6,4'-trihydroxy-7,3'-dimethoxyflavone, and quercetin displayed a mixed type inhibition of 26S by Lineweaver-Burk plots analysis. Furthermore, 5,6,3',4'-tetrahydroxy-7-methoxyflavone is found to have a higher inhibitory effect on 26S proteasome activities and is the only flavonoid to inhibit all three peptidase activities, whereas the inhibition of flavonoids was not affected by ubiquitin-induced stimulation of the three peptidase activities of 26S proteasome; 5,6,3',4'-tetrahydroxy-7-methoxyflavone inhibited 75% casein degradation. These results suggest that both the 6-hydroxy and 7-methoxy positions of the flavone may play an important role in targeting 26S activity.

Objective: We sought to determine whether the MNA (Mininutritional Assessment) would be an effective tool for assessing the nutritional status of patients undergoing hemodialysis., Design: Purposive sampling., Setting: Two hospital-managed hemodialysis centers in central Taiwan., Patients: Subjects were 95 ambulatory patients older than 40 years without acute diseases or infections who had received hemodialysis treatment at the center for longer than 30 days., Methods: Each subject was interviewed with a structured questionnaire to elicit basic personal data and health- and lifestyle-related information and answered questions on the Council on Nutrition Appetite Questionnaire and MNA. Serum biochemical data were obtained from their routine measurement. The nutritional status of each subject was graded with two modified MNA versions. MNA-TI adopted population-specific anthropometric cut-points, and MNA-TII further had the body mass index question deleted from the scale and question scores adjusted but maintained the same 30 total points., Results: Based on the strength of correlation with the key nutrition-related parameters including appetite status, serum creatinine, percent weight loss, hospital length of stay, number of prescribed drugs, and hemodialysis time, both modified MNA versions reflected the nutritional status of Taiwanese hemodialysis patients better than the Council on Nutrition Appetite Questionnaire. Serum albumin did not reflect nutritional status well in these hemodialysis patients. MNA-TI predicted 26.4% and MNA-TII predicted 29.5% of hemodialysis patients were either malnourished or at risk of malnutrition., Conclusions: Both versions of the modified MNA are effective in assessing the nutritional status of hemodialysis patients. MNA-TII without body mass index is a significant improvement for hemodialysis patients because body weights fluctuate significantly between dialysis sessions.

Aim and Objectives: To determine whether a modified version of the Mini Nutritional Assessment (MNA) without body mass index (BMI) can effectively identify individuals at risk of malnutrition among patients with neuropsychiatric disorders., Background: Neuropsychiatric patients have an additional risk of nutritional disorder due to functional impairments and drug effects. However, their nutritional status is generally neglected. It is important to find a tool that is simple, easy to use and non-invasive., Design: The study involved 105 patients in the acute phase of confirmed neuropsychiatric disorders in an area hospital. All subjects were cognitively able to have effective verbal communication., Method: The study included serum biochemical and anthropometric measurements and an on-site, in-person interview using a structured questionnaire to elicit personal data, health condition and answers to questions in the MNA. Subjects' nutritional statuses were graded with a MNA that adopted population-specific anthropometric cut-off points or one further with the BMI question removed and its assigned score redistributed to other anthropometric questions., Results: Both versions of the modified MNA effectively graded the nutritional status of neuropsychiatric patients and showed good correlations with the major nutritional indicators such as BMI, calf circumference and the length of hospital stay., Conclusions: The MNA can effectively assess the nutritional status of neuropsychiatric patients and enhance timely detection and intervention of their nutritional disorders. A modified MNA without the BMI question can maintain the full functionality of the tool. The version does not require weight and height measurements and thus will enhance the usefulness of the instrument., Relevance to Clinical Practice: Neuropsychiatric patients are a high-risk group of nutritional disorders. The MNA, especially the one without BMI, has the potential to improve professional efficiency of the primary care workers.

Objective: To analyze whether magnetic resonance imaging (MRI) findings can be used to predict therapeutic responses to cervical epidural steroid injections (CESI) in patients with cervical radiculopathy., Design: Retrospective single-center study. Patients were categorized by presence or absence of four types of cervical MRI findings (disk herniation, nerve root compromise, neuroforaminal stenosis, and central canal stenosis). Thus, for each of the four types of MRI findings, there was a positive-MRI arm and a negative-MRI arm. Severities of these MRI findings were also noted. The treatment outcome variable was the change in Neck Disability Index (NDI) score after CESI. For each of the four MRI finding types, t test was done to see if there was any significant difference in the outcome between positive-MRI arm and negative-MRI arm. Then, we repeated the same analyses after adjusting the arms to include only those of "major" radiologic severity in the positive-MRI arms., Results: After inclusion and exclusion criteria were applied, 32 patients were included. Only the presence, vs. absence, of central canal stenosis was associated with significantly superior therapeutic response to CESI (mean NDI improvement of 14.14 in positive-MRI arm, vs. mean NDI worsening of 2.11 in negative-MRI arm; P = 0.012). After controlling for other MRI finding types, the statistical significance still held for central canal stenosis (beta = -0.438; P = 0.022). When only the patients with MRI findings of major severities were considered as positive results, central canal stenosis was still the only finding type associated with significantly superior outcome (mean NDI improvement of 23.00 in positive-MRI arm, vs. mean NDI improvement of 0.85 in negative-MRI arm; P = 0.002). Controlling for other MRI finding types, the statistical significance still held for central canal stenosis (beta = -0.534; P = 0.002)., Conclusions: Patients with central canal stenosis achieved significantly better functional outcome after CESI than those without. Hence, the MRI finding of central canal stenosis is a potential indication that CESI may be merited.

Defensins are antimicrobial peptides important to innate host defense. In addition to their direct antimicrobial effect, defensins modulate immune responses. Increasing evidence indicates that defensins exhibit complex functions by positively or negatively modulating infections of both enveloped and non-enveloped viruses. The effects of defensins on viral infections appear to be specific to the defensin, virus and target cell. Regulation of viral infection by defensins is achieved by multiple mechanisms. This review focuses on the interplay between defensins and viral infections, the mechanisms of action of defensins and the in vivo studies of the role of defensins in viral infections., (Copyright 2009 S. Karger AG, Basel.)

Drugs targeting 26S proteasome as antitumor agents are considered to be important for cancer therapy. Although the active components are yet to be identified, for more than 1000 years, the low-toxicity Panax ginseng has been used in traditional herbal medicine for either treating or preventing cancer. Ginsenosides Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1 are distinct components that can be isolated from P. ginseng C.A. Meyer. In this study 26S proteasome was purified from pig red blood cells, and the activity of the seven isolated ginsenosides was analyzed by proteolysis assay. It was found that ginsenoside Rd inhibited 52.9% the chymotrypsin-like activity of 26S proteasome with an IC(50) value of 107.5 microM when Suc-LLVY-AMC was used as a substrate. Ginsenoside Rd displayed a mixed type inhibition of 26S proteasome when analyzed by Lineweaver-Burk plots of the inhibition kinetics. Unlike ginsenoside Rd, the other ginsenosides showed low inhibitory effect of the chymotrypsin-like activity of 26S proteasome. Seven ginsenosides did not inhibit the trypsin-like and caspase-like activities of 26S when Ac-RLR-AMC or Z-LLE-AMC was used as substrate. These results suggest that ginsenoside Rd is a potential drug for cancer prevention due to its specific 26S proteasome inhibitory effect and known low toxicity. Furthermore, both 3-O-Glc(2)-Glc and 20-O-beta-Glc positions of the ginsenoside may play a role in the inhibitory property of the chymotrypsin-like activity in 26S proteasome.

Sexually transmitted infections (STIs) increase the likelihood of HIV transmission. Defensins are part of the innate mucosal immune response to STIs and therefore we investigated their role in HIV infection. We found that human defensins 5 and 6 (HD5 and HD6) promoted HIV infection, and this effect was primarily during viral entry. Enhancement was seen with primary viral isolates in primary CD4(+) T cells and the effect was more pronounced with R5 virus compared with X4 virus. HD5 and HD6 promoted HIV reporter viruses pseudotyped with vesicular stomatitis virus and murine leukemia virus envelopes, indicating that defensin-mediated enhancement was not dependent on CD4 and coreceptors. Enhancement of HIV by HD5 and HD6 was influenced by the structure of the peptides, as loss of the intramolecular cysteine bonds was associated with loss of the HIV-enhancing effect. Pro-HD5, the precursor and intracellular form of HD5, also exhibited HIV-enhancing effect. Using a cervicovaginal tissue culture system, we found that expression of HD5 and HD6 was induced in response to Neisseria gonorrhoeae (GC, for gonococcus) infection and that conditioned medium from GC-exposed cervicovaginal epithelial cells with elevated levels of HD5 also enhanced HIV infection. Introduction of small interfering RNAs for HD5 or HD6 abolished the HIV-enhancing effect mediated by GC. Thus, the induction of these defensins in the mucosa in the setting of GC infection could facilitate HIV infection. Furthermore, this study demonstrates the complexity of defensins as innate immune mediators in HIV transmission and warrants further investigation of the mechanism by which defensins modulate HIV infection.

Simultaneous avulsions of the tibial tubercle and patellar ligament have been reported, but are rare. We present an 11-year-old boy who was initially diagnosed with a Type IIIA tibial tubercle avulsion fracture after falling off of his bicycle. Intraoperatively, following exposure and fixation of the tibial tubercle fragment, knee range of motion under live fluoroscopy revealed a stationary patella with no tension in the patellar ligament. The incision was extended and a distal avulsion of the patellar ligament from the tibial tubercle was identified, with a small flap of tibial tubercle periosteum remaining attached. Transosseous suture fixation of the avulsed periosteal flap was achieved with 2 No. 5 ethibond sutures placed through connecting drill holes. Postoperatively, the patient was placed in a long leg cast with the knee in extension for 6 weeks. The injury to healed and the patient returned to full activities, although he required 2 courses of physical therapy and 4.5 months to regain full range of motion. Preoperative diagnosis of simultaneous tibial tubercle fracture and patellar ligament avulsion can be difficult. Palpation of the patellar ligament for gaps may not be possible due to a large knee effusion. Most patients do not tolerate quadriceps testing on examination and do not have patella alta on radiographs. In retrospect, a clue to this diagnosis in our patient was the preoperative radiographic finding of multiple calcified fragments below the patella. The calcified fragments likely represent the avulsed tibial tubercle periosteum attached to the distal patellar ligament. In addition, the large tibial tubercle fragment is separated and rotated superiorly, a finding that was also noted in two other similar case reports.

Objective: To present a case report of a patient with an open fracture and severe burns and review the literature., Methods: The patient was treated with intubation, intravenous antibiotics, and debridement and intramedullary nailing for the femur fracture. He later underwent multiple burn excision procedures with allograft and autograft skin coverage. The wound over the fracture was treated with dressing changes. The fracture was treated with nail exchange and bone grafting for atrophic nonunion., Results: The patient was returned to full weightbearing and good function with a fully healed femur., Conclusions: Treatment of open fractures in burn patients should be tailored to the specific needs of the individual; they should be reduced and stabilized via internal fixation at the earliest opportunity and should be managed by minimizing wound colonization through successive debridement, wound care, and consideration of flap coverage.

The rhizomes of the Zingiberaceae family are a vegetable widely used in many Asian countries, and their medicinal functions have been broadly discussed and accepted in many traditional recipes. In this study, 18 species of five genus of Zingiberaceae plants from Taiwan area were collected and analyzed for their functional properties. Methanolic extracts of the plants were analyzed for their total phenol compounds, alpha,alpha-diphenyl-beta-picrylhydrazyl (DPPH) scavenging activity, and reducing power. Antimicrobial activity of these samples was also determined. The results showed that the total phenol compounds of the Alpinia genus averaged 17, 30 mg/g for Curcumas, and the highest, 36.5 mg/g for Vanoverberghia sasakiana. Antioxidant performances were best observed in Vanoverberghia and Hedychium, both 89%, and DPPH scavenging activity followed similar trends. Particularly, Zingiber oligophyllum, considered as a traditional medicinal plant used in Taiwan exhibited low DPPH scavenging activity and reducing power. Most Zingiberaceae plant extracts exhibited antimicrobial activity against all tested food microorganisms. Hedychium and Vanoverberghia, did not show antimicrobial activities on Escherichia coli and Vibrio parahaemolyticus. This study is a positive demonstration of the utility of screening Taiwan's endemic Zingiberaceous plants for their food and medicinal uses.

Background: Lumbar decompressive surgery can be complicated by dural tears, infection, nerve root injury, deep venous thrombosis, and epidural hematoma. However, perioperative complications of multilevel laminectomies in patients with achondroplasia rarely are reported. Our objective was to determine the perioperative complication rates associated with laminectomies in patients with achondroplasia., Methods: We reviewed the medical records of the initial laminectomies for all ninety-eight patients with achondroplasia at our institution, which included twenty-eight patients who had had previous spine surgeries at other institutions, and determined the rates of complications in the following categories: neurologic, pulmonary, cardiovascular, and gastrointestinal complications; intraoperative dural tears; infections at the incision site; and mortality., Results: Sixty (61%) of ninety-eight patients had at least one perioperative complication. By category, these included intraoperative dural tears, which occurred in 37% (thirty-six patients); neurologic complications, in 23% (twenty-three patients); infections at the incision site, in 9% (nine patients); deep venous thrombosis, in 3% (three patients); pulmonary complications, in 3% (three patients); and gastrointestinal complications, in 3% (three patients). The only death was caused by a pulmonary embolism., Conclusions: Perioperative complications are common in patients with achondroplasia undergoing multilevel laminectomies, and the perioperative care team should be aware of these complications when caring for these patients.

Purpose: To evaluate the primary stability of dental implants placed with condensing-osteotome versus drilling-osteotome techniques and to explore peri-implant mircromorphologic consequences of lateral bone condensing., Materials and Methods: The experimental model designed for the study comprised bilateral iliac crests from 3 fresh frozen human cadavers. Two AstraTech dental implants (AstraTech AB, Mölndal, Sweden) were consecutively placed with condensing- and drilling-osteotome techniques in bone with a 10-mm interimplant distance. Six experimental bone sites received a total of 12 implants. Installation torque values (ITVs) and implant stability quotients (ISQs) were measured to quantify primary implant stability. Bone specimens including implants were removed to quantify the peri-implant relative bone volume and bone microstructural parameters in the 1-mm circular vicinity of implants using desktop computed tomography (microCT). The Mann-Whitney U test was used to evaluate the differences in primary implant stability values and microCT data for the surgical placement techniques., Results: ITVs and ISQs were similar for both surgical placement techniques without statistical significance (P > .05). Relative bone volumes around implants placed with the condensing-osteotome technique were significantly (P < .05) higher than those around implants placed with the drilling-osteotome technique. Microstructural parameters, such as trabecular thickness, separation, and number, differed significantly between the 2 osteotome techniques., Conclusions: Managing implant sites with the condensing-osteotome technique results in notable changes in peri-implant bone architecture but might not be as promising in improving primary implant stability compared with the drilling-osteotome technique.

SAMMA, a mandelic acid condensation polymer, exhibits a broad antimicrobial activity against several sexually transmitted pathogens including human immunodeficiency virus (HIV). Here we demonstrated that SAMMA suppressed HIV transmission by dendritic cells (DCs), one of the first target cells for primary infection. The greatest inhibitory effect was achieved when SAMMA was present during the co-culture with target cells. The inhibitory effect of SAMMA on DC-mediated HIV transmission was not due to cytotoxicity. Analysis of the level of DC-associated HIV p24 antigen revealed that SAMMA prevented HIV internalization by DCs when the virus was pre-incubated with the compound. In contrast, pre-incubation of DCs with SAMMA followed by wash-off did not affect the amount of cell-associated HIV p24 antigen. In addition, SAMMA blocked HIV glycoprotein-mediated cell-cell fusion. This study suggests that SAMMA prevents HIV infection through multiple mechanisms.

Background: Osteoporosis may present a risk factor in achievement of osseointegration because of its impact on bone remodeling properties of skeletal phsiology. The purpose of this study was to evaluate micro-morphological changes in bone around titanium implants exposed to mechanical and electrical-energy in osteoporotic rats., Methods: Fifteen 12-week old sprague-dowley rats were ovariectomized to develop osteoporosis. After 8 weeks of healing period, two titanium implants were bilaterally placed in the proximal metaphyses of tibia. The animals were randomly divided into a control group and biophysically-stimulated two test groups with five animals in each group. In the first test group, a pulsed electromagnetic field (PEMF) stimulation was administrated at a 0.2 mT 4 h/day, whereas the second group received low-magnitude high-frequency mechanical vibration (MECHVIB) at 50 Hz 14 min/day. Following completion of two week treatment period, all animals were sacrificed. Bone sites including implants were sectioned, removed en bloc and analyzed using a microCT unit. Relative bone volume and bone micro-structural parameters were evaluated for 144 mum wide peri-implant volume of interest (VOI)., Results: Mean relative bone volume in the peri-implant VOI around implants PEMF and MECHVIB was significantly higher than of those in control (P < .05). Differences in trabecular-thickness and -separation around implants in all groups were similar (P > .05) while the difference in trabecular-number among test and control groups was significant in all VOIs (P < .05)., Conclusion: Biophysical stimulation remarkably enhances bone volume around titanium implants placed in osteoporotic rats. Low-magnitude high-frequency MECHVIB is more effective than PEMF on bone healing in terms of relative bone volume.

The present study describes an ultrasensitive protein biochip that employs nanogap electrodes and self-assembled nanoparticles to electrically detect protein. A bio-barcode DNA technique amplifies the concentration of target antigen at least 100-fold. This technique requires the establishment of conjugate magnetic nanoparticles (MNPs) and gold nanoparticles (AuNPs) through binding between monoclonal antibodies (2B2), the target antigen, and polyclonal antibodies (GP). Both GP and capture ssDNA (single-strand DNA) bonds to bio-barcode ssDNA are immobilized on the surface of AuNPs. A denature process releases the bio-barcode ssDNAs into the solution, and a hybridization process establishes multilayer AuNPs over the gap surface between electrodes. Electric current through double-layer self-assembled AuNPs is much greater than that through self-assembled monolayer AuNPs. This significant increase in electric current provides evidence that the solution contains the target antigen. Results show that the protein biochip attains a sensitivity of up to 1 pg/ microL.

Obesity is a widespread and potentially serious health problem in individuals with achondroplasia. In addition to obesity, such individuals commonly experience lumbar spinal stenosis. Although laminectomies have long been the method of choice for thorough lumbar decompression, to the best of our knowledge, the relationship between body mass index (BMI) and surgical outcomes after laminectomy in patients with achondroplasia has not been investigated in a large patient population. Through a retrospective medical record review of the initial laminectomies of all individuals with achondroplasia at our institution for whom adequate records were available, we evaluated the influence of BMI on surgical outcomes after laminectomy via four criteria: (1) change in Rankin score; (2) change in walking distance; (3) intra-operative and post-operative complications; and (4) need for subsequent revision laminectomies. The 49 individuals (mean BMI: 31.5 kg/m(2) at the time of laminectomy) were stratified into the standard BMI categories: normal (<25 kg/m(2)), overweight (25-29.9 kg/m(2)), obese Class I (30-34.9 kg/m(2)), and a combined obese Class II (35-39.9 kg/m(2)) and obese Class III (>40 kg/m(2)). Our analysis indicated that there was no statistical difference between these groups in terms of the four criteria. When the study group was categorized into non-obese (normal weight and overweight) and obese (obese Classes I and II/III) groups, there was also no statistically significant improvement difference in terms the same four criteria. Our data suggest that obesity does not pose additional risks to laminectomy in individuals with achondroplasia.

Genistein, one of the major isoflavones, potently inhibits the growth and metastasis of breast cancer. However, the precise molecular mechanism in metastasis inhibition is not clear. We investigated the effect of genistein in HCC1395 cells, a cell line derived from an early-stage primary breast cancer. Genistein dose dependently both decreased cell viability and inhibited the invasion potential. We used human oligonucleotide microarrays to determine the gene expression profile altered by genistein treatment. TFPI-2, ATF3, DNMT1, and MTCBP-1, which inhibit invasion and metastasis, were upregulated, and MMP-2, MMP-7, and CXCL12, which promote invasion and metastasis, were downregulated. We used quantitative real-time polymerase chain reaction to verify the microarray data at the mRNA level. We conclude that genistein-induced alternations of gene expression involving metastasis may be exploited for devising chemopreventive and therapeutic strategies, particularly for early-stage breast cancer.

The 26S proteasome degrades polyubiquitinated proteins by an energy-dependent mechanism. Here we define multiple roles for ATP in 26S proteasome function. ATP binding is necessary and sufficient for assembly of 26S proteasome from 20S proteasome and PA700/19S subcomplexes and for proteasome activation. Proteasome assembly and activation may require distinct ATP binding events. The 26S proteasome degrades nonubiquitylated, unstructured proteins without ATP hydrolysis, indicating that substrate translocation per se does not require the energy of hydrolysis. Nonubiquitylated folded proteins and certain polyubiquitylated folded proteins were refractory to proteolysis. The latter were deubiquitylated by an ATP-independent mechanism. Other folded as well as unstructured polyubiquitylated proteins required ATP hydrolysis for proteolysis and deubiquitylation. Thus, ATP hydrolysis is not used solely for substrate unfolding. These results indicate that 26S proteasome-catalyzed degradation of polyubiquitylated proteins involves mechanistic coupling of several processes and that such coupling imposes an energy requirement not apparent for any isolated process.

Osseointegrated implants can be applied to facilitate retention, stability, and support for facial and intraoral prostheses used to restore head and neck defects. At the University of California, Los Angeles, Maxillofacial Prosthetics Clinic, retrospective studies have indicated that in nonirradiated maxillectomy patients, implant survival rates are 82.6 percent. In mandibles reconstructed with fibula free flaps, survival rates are 94.6 percent. Similarly, high implant survival rates have been observed for most sites used to support facial prostheses. Cumulative six-year survival rates for auricular sites exceed 95 percent and for floor of nose sites, success rates exceed 87 percent. However, survival rates are low (53 percent) for implants placed in the frontal bone for retention of orbital prostheses and even lower for irradiated bone sites ranging from 63 percent in the maxilla to 27 percent in the orbit.

Hypodontia is a relatively rare occurrence that can have a significant impact on treatment planning for those patients with the condition. This paper will describe the forms of hypodontia, as well as associated dental issues. Treatment planning considerations for children, adolescents, and adults will be presented.

Purpose: The capability of micro-computed tomography (microCT) for quantitative analysis of peri-implant bone has not been previously addressed. This study aimed to establish and validate a method to use this technique for 3-dimensional bone-implant integration profiling., Materials and Methods: Unthreaded cylindric implants with a dual acid-etched surface were placed into the right femurs of 7 Sprague-Dawley rats. Two weeks postimplantation, the femurs were harvested and measured with a desktop micro-tomographic scanner with an isotropic resolution of 8 microm. To validate the microCT outcome, ground histologic sections and corresponding CT slices were compared with respect to bone morphometry., Results: Bone-implant integration profiles assessed by microCT revealed that the percentage of cancellous bone gradually increased with proximity to the implant surface, while the percentage of cortical bone was not affected by proximity to the implant. Using the optimized segmentation threshold, the bone configuration in the microCT images corresponded to that observed in the histologic sections. The correlation between microCT and histology was significant for cortical (r = 0.65; P < .05) and cancellous bone (r = 0.92; P < .05) at distances of 24 to 240 microm from the implant surface, but no significant correlation was found for the area from 0 to 24 microm from the surface., Discussion and Conclusion: The results support the usefulness of microCT assessment as a rapid, nondestructive method for 3-dimensional bone ratio measurements around implants, which may provide new perspectives for osseointegration research. Further study is necessary, however, to address the inherent metallic halation artifact, which potentially confounds peri-implant bone assessment.

Purpose: The objective of this biomechanical study was to explore the effect of bone micro-morphology on initial intraosseous stability of implants with different designs., Material and Methods: Straumann and Astra Tech dental implants were placed into anterior and posterior regions of completely edentulous maxilla and mandible of a human cadaver. Experiments were undertaken to quantify initial implant stability and bone micro-morphology. Installation torque values (ITVs) and implant stability quotients (ISQs) were measured to determine initial intraosseous implant stability. For quantification of relative bone volume and micro-architecture, sectioned implant-bone and bone core specimens of each implant placement site were consecutively scanned and trabecular bone was analyzed in a micro-computed tomography (micro-CT) unit. Experimental outcomes were evaluated for correlations among implant designs, initial intraosseous implant stability and bone micro-structural parameters., Results: ITVs correlated higher with bone volume fraction (BV/TV) than ISQs, at 88.1% and 68.9% levels, respectively. Correlations between ITVs and micro-morphometric parameters were significant at the 95% confidence level (P<0.05) while ISQs were not. Differences in ITVs, ISQs and BV/TV data in regards to implant designs used were not significant at the 95% confidence level (P>0.05)., Conclusion: Bone micro-morphology has a prevailing effect over implant design on intraosseus initial implant stability, and ITV is more sensitive in terms of revealing biomechanical properties at the bone-implant interface in comparison with ISQ.

Mallet fingers are difficult to treat, especially in surgeons, who need to sterilize their hands continuously and who have constant strains placed on their fingers. We successfully treated a nondominant, fifth-digit, nonbony mallet finger in a surgical resident with a splint method composed of a bent, semitubular, small-fragment plate and Steri-strips (3-M, St. Paul, MN). This splinting method allowed the resident to continue performing surgeries while the injury healed.

Defensins are small antimicrobial peptides that are produced by leukocytes and epithelial cells, and that have an important role in innate immunity. Recent advances in understanding the mechanisms of the antiviral action(s) of defensins indicate that they have a dual role in antiviral defence, acting directly on the virion and on the host cell. This Review focuses on the antiviral activities and mechanisms of action of mammalian defensins, and on the clinical relevance of these activities. Understanding the complex function of defensins in innate immunity against viral infection has implications for the prevention and treatment of viral disease.

Purpose: To compare the dimensional accuracy of implant framework castings from an argon vacuum casting machine with those from a centrifugal casting machine., Materials and Methods: Three 4 x 10-mm external hex-type implants (3i/Implant Innovations) were embedded in an acrylic resin block 7 mm apart, with a 2 mm offset of the middle implant. Eight reference points were marked on the implant collars. Twenty implant bar frameworks were waxed with UCLA abutments, invested with a ringless system, and subjected to the same thermal cycle. Ten wax patterns were cast in gold alloy using an oxygen-propane torch and centrifugal casting system; 10 were cast using an argon vacuum casting machine (KDF; Denken). The White 1-screw technique was applied after sequentially tightening the mesial and distal abutment screws to 10 Ncm. Fit of the implant framework castings was evaluated by measuring the marginal opening between the casting and implant at the reference points. These measurements were averaged and statistically compared for differences., Results: The mean marginal openings at the most distant measuring locations from the tightened retaining screw at location 1 was between 44 to 48 microm for the centrifugal system compared to between 28 to 32 mm for KDF (P < .01). For screws tightened at location 3, the mean marginal openings at the most distant measuring locations were between 40 to 51 mm for the centrifugal system compared to between 27 to 29 microm for KDF (P < .01)., Discussion: In comparison with the centrifugal casting and oxygen-propane system, the argon vacuum system was more accurate and user friendly and less technique-sensitive., Conclusion: The argon vacuum casting machine tested produced more accurate, better fitting implant-supported prosthesis frameworks than a conventional centrifugal casting system. The "1-screw" method of evaluating casting fit was most effective when either of the prostheses' end screws were tightened.

Statement of Problem: Facial defects secondary to the treatment of neoplasms, congenital malformations, and trauma result in multiple functional and psychosocial difficulties. Prosthetic rehabilitation attempts to restore these facial disfigurements and may improve the level of function and self-esteem for these patients. However, a limited number of studies have evaluated the change in perceived quality of life after maxillofacial prosthetic rehabilitation., Purpose: The purpose of this study was to evaluate patients' perceptions of treatment with adhesive-retained and implant-retained facial prostheses and to assess differences in overall satisfaction with these 2 types of treatments., Material and Methods: In this study, a questionnaire with 28 items was administered for evaluation of perceptions of appearance, comfort, fit and irritation, reliability of retention, frequency of wear, ease of placement and removal, level of self-consciousness, and value of treatment. Subjects were categorized into 2 groups: adhesive-retained group (n=16) and implant-retained group (n=19). Comparisons were made for each item in the questionnaire using Fisher exact tests (alpha=.05)., Results: The implant group reported higher positive ratings on all 28 questionnaire items when compared with the adhesive group. Statistically significant (P<.05) differences between the implant and adhesive groups were noted for ease of placement and removal, frequency of wear at home, and quality of retention during various activities, such as home chores and when perspiring or sneezing/coughing., Conclusion: The implant-retained facial prosthesis offers significant enhancement over an adhesive-retained prosthesis with respect to ease of use and retention during a variety of daily activities, resulting in greater use of the prosthesis.

Erythrocyte spectrin contains E2/E3 ubiquitin conjugating/ligating activity in its alpha subunit. Ankyrin is a target of spectrin's E2/E3 ubiquitin conjugating/ligating activity in vitro and in vivo. We compare the ubiquitination levels of ankyrin mediated by control and sickle cell spectrin using a biotinylated ubiquitin cell-free assay. Sickle cell spectrin has diminished ability to transfer ubiquitin from an intermediate spectrin-ubiquitin thioester adduct (alpha' spectrin) to ankyrin, which may be due to glutathiolation of spectrin's E2 and/or E3 active site cysteines. There is also a diminished ability of the sickle cell ankyrin to serve as target of spectrin's E2/E3 activity, probably due to oxidative damage to ankyrin. A direct correlation exists between the alpha'/alpha spectrin ratio and spectrin's ability to ubiquitinate ankyrin. There is also an inverse correlation between severity of the disease and the alpha'/alpha spectrin ratio in SS erythrocytes. These results suggest that reduced spectrin E2/E3 activity is an important determinant of sickle cell severity.

Alpha-defensins are abundant antimicrobial peptides in polymorphonuclear leukocytes and play an important role in innate immunity. We have previously shown that alpha-defensin-1 can inhibit HIV-1 replication following viral entry. Here we examined the molecular mechanism(s) of alpha-defensin-1-mediated HIV-1 inhibition. Alpha-defensin-1 had a direct effect on HIV-1 virions at a low MOI in the absence of serum. The direct effect on HIV-1 virions was abolished by the presence of serum or an increase in virus particles. Studying the kinetics of the HIV life cycle revealed that alpha-defensin-1 inhibited steps following reverse transcription and integration. Analysis of PKC phosphorylation in primary CD4+ T cells in response to alpha-defensin-1 indicated that alpha-defensin-1 inhibited PKC activity. Pretreatment of infected CD4+ T cells with a PKC activator, bryostatin 1, partially reversed alpha-defensin-1-mediated HIV inhibition. Like alpha-defensin-1, the PKC isoform-selective inhibitor Go6976 blocked HIV-1 infection in a dose-dependent manner. Furthermore, kinetic studies and analysis of HIV-1 products indicated that alpha-defensin-1 and Go6976 blocked HIV-1 infection at similar stages in its life cycle, including nuclear import and transcription. Taken together, our studies demonstrate that, in the absence of serum, alpha-defensin-1 may act directly on the virus, but, in the presence of serum, its effects are on the cell, where it inhibits HIV-1 replication. At least 1 of the cellular effects associated with HIV inhibition is interference with PKC signaling in primary CD4+ T cells. Studying the complex function of alpha-defensin-1 in innate immunity against HIV has implications for prevention as well as therapeutics.

The expressed sequence tag (EST) effort in Toxoplasma gondii has generated a substantial amount of gene information. To exploit this valuable resource, we chose to study tgd057, a novel gene identified by a large number of ESTs that otherwise show no significant match to known sequences in the database. Northern analysis showed that tgd057 is transcribed in this tachyzoite. The complete cDNA sequence of tgd057 is 1169 bp in length. Sequence analysis revealed that tgd057 possibly adopts two polyadenylation sites, utilizes the fourth in-frame ATG for translation initiation, and codes for a secretory protein. The longest open reading frame for the tgd057 gene was cloned and expressed as a recombinant protein (rd57) in Escherichia coli. Western analysis revealed that serum against rd57 recognized a molecule of ~21 kDa in the tachyzoite protein extract. This suggests that the tgd057 gene is expressed in vivo in the parasite.

Mammalian defensins are small cationic antimicrobial peptides predominantly found in leukocytes and epithelial cells engaged in host defense. These peptides act as effector molecules in innate immunity as well as regulators in adaptive immunity. Increasing evidence indicates that defensins are effective inhibitors of HIV-1. While the level of defensins in HIV-1 infected individuals has not been determined, neutropenia and neutrophil dysfunction associated with HIV disease progression may result in altered alpha-defensin production. This review provides an overview of the structure and function of defensins, and focuses on the anti-HIV-1 activity of defensins and the mechanism of this activity. Although many questions remain, studying the complex function of defensins in innate immunity against HIV has implications for our further understanding of disease progression and for the development of novel approaches to prevention and therapy.

The mechanical behavior of bone is a critical factor in the attainment and maintenance of osseointegration. Current procedures and classifications in assessing bone quality have limitations. Micro-computed tomography (microCT) is a new method to image and quantify bone with very high resolution. This study was aimed to analyze cadaveric maxillary and mandibular trabecular bone with 3-D morphometric data acquired through microCT and correlate with conventional bone assessment methods. Moderately resorbed edentulous maxilla and mandible from a human cadaver were scanned in the conventional CT unit with opaque markers indicating specific anterior and posterior sites. These sites were then sectioned and standard periapical radiographs were obtained. Bone cores were harvested from the sectioned sites and scanned in the microCT unit. Bone density values based on the Hounsfield scale ranged from 51 to 529 in the mandible and 186 to 389 in the maxilla, anterior sites being higher in both. Periapical images did not yield distinct differences. 3-D morphometric analysis in microCT produced a range of values with anterior specimens being favorable: bone volume density (0.12-0.291), trabecular thickness (0.12-0.16 mm), trabecular separation (0.46-0.82 mm), trabecular number (1.08-2.071/mm) and structural model index (0.29-1.27). General agreement between bone density and microCT indices was noted; however, subtle differences have to be studied with larger samples. This preliminary study suggests that the understanding of mechanical competence of trabecular bone might reveal further information about the prognosis of implant therapy, advancements in implant design, surgical techniques and grafting.