Green tea extract supplementation ameliorates CCl4-induced hepatic oxidative stress, fibrosis, and acute-phase protein expression in rat

Background/PurposeWe evaluated the long-term effects of green tea extract (GTE) supplementation on oxidative stress, biliary acute phase protein expression, and liver function in CCl4-induced chronic liver injury.MethodsWe evaluated the antioxidant activity of GTE in comparison with those of vitamin C, vitamin E, and β-carotene in vitro by using an ultrasensitive chemiluminescence analyzer. Chronic liver injury was induced by intraperitoneally administering carbon tetrachloride (CCl4) (1mL/kg body weight, twice weekly) to female Wistar rats for 8 weeks. The effects of low (4mg/kg body weight per day) and high (20mg/kg body weight per day) doses of intragastric GTE on CCl4-induced liver dysfunction and fibrosis were examined by measuring the bile and blood reactive oxygen species levels and biochemical parameters by using Western blot and two-dimensional polyacrylamide gel electrophoresis techniques.ResultsGTE has greater scavenging activity against O2–, H2O2, and Hypochlorous acid (HOCl) in vitro than vitamin C, vitamin E, and β-carotene do. In vivo, CCl4 markedly increased bile and blood reactive oxygen species production, lipid accumulation, number of infiltrated leukocytes, fibrosis, hepatic hydroxyproline content, and plasma alanine aminotransferase and aspartate aminotransferase activities, and reduced plasma albumin levels. Two-dimensional polyacrylamide gel electrophoresis revealed that CCl4 increased the acute-phase expression of six biliary proteins and decreased hepatic B-cell lymphoma 2 (Bcl-2), catalase, and CuZn superoxide dismutase protein expression. GTE supplementation attenuated CCl4-enhanced oxidative stress, levels of biochemical parameters, pathology, and acute-phase protein secretion, and preserved antioxidant/antiapoptotic protein expression.ConclusionGTE supplementation attenuates CCl4-induced hepatic oxidative stress, fibrosis, acute phase protein excretion, and hepatic dysfunction via the antioxidant and antiapoptotic defense mechanisms

Gene expression profiling of breast cancer survivability by pooled cDNA microarray analysis using logistic regression, artificial neural networks and decision trees

BACKGROUND: Microarray technology can acquire information about thousands of genes simultaneously. We analyzed published breast cancer microarray databases to predict five-year recurrence and compared the performance of three data mining algorithms of artificial neural networks (ANN), decision trees (DT) and logistic regression (LR) and two composite models of DT-ANN and DT-LR. The collection of microarray datasets from the Gene Expression Omnibus, four breast cancer datasets were pooled for predicting five-year breast cancer relapse. After data compilation, 757 subjects, 5 clinical variables and 13,452 genetic variables were aggregated. The bootstrap method, Mann–Whitney U test and 20-fold cross-validation were performed to investigate candidate genes with 100 most-significant p-values. The predictive powers of DT, LR and ANN models were assessed using accuracy and the area under ROC curve. The associated genes were evaluated using Cox regression. RESULTS: The DT models exhibited the lowest predictive power and the poorest extrapolation when applied to the test samples. The ANN models displayed the best predictive power and showed the best extrapolation. The 21 most-associated genes, as determined by integration of each model, were analyzed using Cox regression with a 3.53-fold (95% CI: 2.24-5.58) increased risk of breast cancer five-year recurrence… CONCLUSIONS: The 21 selected genes can predict breast cancer recurrence. Among these genes, CCNB1, PLK1 and TOP2A are in the cell cycle G2/M DNA damage checkpoint pathway. Oncologists can offer the genetic information for patients when understanding the gene expression profiles on breast cancer recurrence

Clinical and molecular characterization of a transmitted reciprocal translocation t(1;12)(p32.1;q21.3) in a family co-segregating with mental retardation, language delay, and microcephaly

<p>Abstract</p> <p>Background</p> <p>Chromosome translocation associated with neurodevelopmental disorders provides an opportunity to identify new disease-associated genes and gain new insight into their function. During chromosome analysis, we identified a reciprocal translocation between chromosomes 1p and 12q, t(1; 12)(p32.1; q21.3), co-segregating with microcephaly, language delay, and severe psychomotor retardation in a mother and her two affected boys.</p> <p>Methods</p> <p>Fluorescence in situ hybridization (FISH), long-range PCR, and direct sequencing were used to map the breakpoints on chromosomes 1p and 12q. A reporter gene assay was conducted in human neuroblastoma (SKNSH) and Chinese hamster ovary (CHO) cell lines to assess the functional implication of the fusion sequences between chromosomes 12 and 1.</p> <p>Results</p> <p>We determined both breakpoints at the nucleotide level. Neither breakpoint disrupted any known gene directly. The breakpoint on chromosome 1p was located amid a gene-poor region of ~ 1.1 Mb, while the breakpoint on chromosome 12q was located ~ 3.4 kb downstream of the ALX1 gene, a homeobox gene. In the reporter gene assay, we discovered that the fusion sequences construct between chromosomes 12 and 1 had a ~ 1.5 to 2-fold increased reporter gene activity compared with the corresponding normal chromosome 12 sequences construct.</p> <p>Conclusion</p> <p>Our findings imply that the translocation may enhance the expression of the ALX1 gene via the position effect and result in the clinical symptoms of this family. Our findings may also expand the clinical phenotype spectrum of ALX1-related human diseases as loss of the ALX1 function was recently reported to result in abnormal craniofacial development.</p

Numerical and experimental study of horizontal pneumatic transportation of spherical and low-aspect-ratio cylindrical particles

The work presented in this paper was carried out as part of the PARDEM project. The overall aim was to quantify the predictive capability of a coupled CFD-DEM approach to simulating the horizontal pneumatic conveying of spherical and low-aspect-ratio non-spherical particles. Carefully controlled experiments were carried out in a 6.5 m long, 0.075 m diameter horizontal conveying line with the aid of the laser Doppler anemometry (LDA). Three different sizes of spherical glass beads, ranging from 0.8 mm to 2 mm and low-aspect-ratio cylindrical shaped particle of size 1 × 1.5 mm were employed. Simulations of the experiments were performed using a two-way coupled computational fluid dynamics and discrete element method (CFD-DEM) implemented in the commercial software FLUENT-EDEM in an Eulerian–Lagrangian framework. Experimental and simulation results of gas and particle velocities for particle laden flow with spherical particles were compared, showing that the CFD-DEM method could capture the experimental trends. However, quantitative discrepancies between simulation and experimental results were observed. Further modelling of low-aspect-ratio cylindrical particles was conducted using a multi-sphere model to represent cylindrical particles in the DEM code. Drag equations were modified in the code to take the effect of particle shape into account. The simulation results of mean axial particle velocity agreed reasonably well with a maximum of 25% discrepancy when compared to experimental measurements using the LDA technique. The discrepancies between simulation and experimental results were attributed to the selected drag model, mesh size and lack of an appropriate mesh interpolation scheme in the selected code

Identification of Critical Amino Acids in an Immunodominant IgE Epitope of Pen c 13, a Major Allergen from Penicillium citrinum

Background: Pen c 13, identified as a 33-kDa alkaline serine protease, is a major allergen secreted by Penicillium citrinum. Detailed knowledge about the epitopes responsible for IgE binding would help inform the diagnosis/prognosis of fungal allergy and facilitate the rational design of hypoallergenic candidate vaccines. The goal of the present study was to characterize the IgE epitopes of Pen c 13. Methodology/Principal Findings: Serum samples were collected from 10 patients with mold allergy and positive Pen c 13 skin test results. IgE-binding epitopes on rPen c 13 were mapped using an enzymatic digestion and chemical cleavage method, followed by dot-blotting and mass spectrometry. A B-cell epitope-predicting server and molecular modeling were used to predict the residues most likely involved in IgE binding. Theoretically predicted IgE-binding regions were further confirmed by site-directed mutagenesis assays. At least twelve different IgE-binding epitopes located throughout Pen c 13 were identified. Of these, peptides S16 (A 148 –E 166) and S22 (A 243 –K 274) were recognized by sera from 90 % and 100 % of the patients tested, and were further confirmed by inhibition assays. Peptide S22 was selected for further analysis of IgE-binding ability. The results of serum screening showed that the majority of IgE-binding ability resided in the C-terminus. One Pen c 13 mutant, G270A (T 261 –K 274), exhibited clearly enhanced IgE reactivity, whereas another, K274A, exhibited dramatically reduced IgE reactivity

Lower Urinary Tract Symptoms in Female Patients with Rheumatoid Arthritis

Objective: Patients with autoimmune diseases such as systemic lupus erythematosus ( SLE) and Sjogren's syndrome ( SS) are associated with an increased severity of lower urinary tract symptoms ( LUTS). Recent surveys also reveal that rheumatoid arthritis ( RA) is prevalent in patients with interstitial cystitis ( IC). Therefore, we have investigated LUTS in patients with RA. Methods: A total of 198 female patients with RA, aged 40 years or older, from the rheumatology outpatient clinic completed this prospective study. The American Urological Association Symptom Index ( AUASI) score was used to assess the severity of LUTS and the O'Leary-Sant Symptom Index ( ICSI) was used to evaluate IC-like urinary symptoms in these patients, which were compared to those of 679 age-matched controls. The possible associations of clinical parameters with LUTS were also explored. Results: The Mean AUASI score and the percentage of individuals reporting severe LUTS ( AUASI score >= 20) or IC-like urinary symptoms ( ICSI score >= 12) showed no significant differences between the RA and control groups. However, in the RA group multivariate regression analyses identified patients with secondary SS ( n=21) to be associated with a significantly higher AUASI score ( p=0.007) and a higher percentage of severe LUTS ( p= 0.02); these were also significantly higher than those of the control group ( p=0.02 and p=0.01, respectively). Conclusion: Patients with RA have similar urinary complaints when compared to controls . However, those with secondary SS have a greater severity of LUTS, a finding similar to that observed in patients with primary SS

Immunoglobulin M and G Immunoblots in the Diagnosis of Parvovirus B19 Infection

Background and purpose: To identify parvovirus B19 infection by means of immunoglobulin (Ig) G and IgM immunoblots among immunocompetent patients who tested negative or had low- titer B19 IgM antibodies in enzyme-linked immunosorbent assays (ELISA). Methods: Serum samples were obtained from 20 patients with parvovirus B19 infection. Another 130 study subjects presumed to be without B19 infection (40 medical personnel and 90 prisoners) were also included. All sera from the patient and study groups tested positive for IgG or IgM with ELISA and were further evaluated using the immunoblot method. Detection of B19 DNA by nested polymerase chain reaction (PCR) was also performed on IgG and IgM positive sera. Results: IgM immunoblots disclosed one false positive IgM ELISA result in the patient group and three false positive results in the study group. In the patient group, four patients were in the latter stage of antibody response to B19 infection as suggested by tie low titer of anti-B19 IgM, incomplete IgM immunoblots, with only a weak viral capsid protein VP-N reaction band, and fading but still strong reaction bands on Ige immunoblots. Strong reaction bands on IgG immunoblots comparable to these foul patients were found in three of the 130 study group sera. Furthermore , B19 DNA was detected in three of the four patients and one of the three study subjects by means of nested-PCR. A serum sample from one study subject showed strong IgG but no IgM reactivity to viral capsid protein VP 2 ; nested PCR identified B19 DNA in this serum sample. Conclusions: Immunoblots and nested PCR should be applied in the diagnosis of B19 infection for patients with low-titer anti-B19 IgM tested by means of ELISA. For diagnosis of B19 infections in certain clinical entities such as chronic arthritis of recent onset and hydrops fetalis, B19 IgM antibodies may have disappeared hut B19 infection can still be recognized by the intensity of the reaction bands on IgG immunoblots. The correlation between chronic B19 infection and persistence of antilinear VP2 epitopes requires further study

Pilocarpine Hydrochloride for the Treatment of Xerostomia in Patients with Sjögren's Syndrome in Taiwan—A Double-blind, Placebo-controlled Trial

Sjögren's syndrome (SS) is characterized by diminished exocrine secretions with the resultant symptoms of dry mouth and dry eye. As genetic predisposition and ethnicity may alter the effectiveness of drug treatment, evaluation of the efficacy and safety of the secretagogue pilocarpine hy-drochloride in the treatment of xerostomia in patients with SS in different populations is needed. Methods: Forty-four patients with SS were enrolled in this double-blind, placebo-controlled trial. Patients were randomized to receive 5 mg pilocarpine (Salagen) or placebo tablet four times daily for 12 weeks. Global evaluation and subjective responses of patients were assessed by questionnaires with visual analog scales and categorical checkboxes. Saliva production was also measured by modified Saxon's test. Results: Pilocarpine treatment significantly improved global assessment of dry mouth, symptoms associated with dry mouth (mouth comfort, ability to sleep and ability to speak), and saliva production compared to placebo. The drug was well tolerated and the most common adverse effect was sweating (5/23, 21.7%) resulting from the muscarinic agonist action of the drug. No serious drug-related adverse effect was found in this study. Conclusion: The results of this study suggest that therapy with 5 mg pilocarpine four times daily is effective, safe and well tolerated for the relief of oral symptoms in patients with SS in Taiwan

Identification of Ndfip1 as a novel negative regulator for spatial memory formation associated with increased ubiquitination of Beclin 1 and PTEN

Long-term memory formation requires de novo RNA and protein synthesis. By using the differential display-polymerase chain reaction strategy, we have presently identified the Nedd4 family interacting protein 1 (Ndfip1) cDNA fragment that is differentially expressed between the slow learners and the fast learners from the water maze learning task in rats. Further, the fast learners show decreased Ndfip1 mRNA and protein expression levels than the slow learners. Spatial training similarly decreases the Ndfip1 mRNA and protein expression levels. Conversely, the Ndfip1 conditional heterozygous (cHet) mice show enhanced spatial memory performance compared to the Ndfip1flox/WT control mice. Result from co-immunoprecipitation experiment indicates that spatial training decreases the association between Ndfip1 and the E3 ubiquitin ligase Nedd4 (Nedd4-1), and we have shown that both Beclin 1 and PTEN are endogenous ubiquitination targets of Nedd4 in the hippocampus. Further, spatial training decreases endogenous Beclin 1 and PTEN ubiquitination, and increases Beclin 1 and PTEN expression in the hippocampus. On the other hand, the Becn1 conditional knockout (cKO) mice and the Pten cKO mice both show impaired spatial learning and memory performance. Moreover, the expression level of Beclin 1 and PTEN is higher in the Ndfip1 cHet mice compared with the Ndfip1flox/WT control mice. Here, we have identified Ndfip1 as a candidate novel negative regulation for spatial memory formation and this is associated with increased ubiquitination of Beclin 1 and PTEN in the hippocampus