CD44 variant‐dependent redox status regulation in liver fluke‐associated cholangiocarcinoma: A target for cholangiocarcinoma treatment

Expression of CD44, especially the variant isoforms (CD44v) of this major cancer stem cell marker, contributes to reactive oxygen species (ROS) defense through stabilizing xCT (a cystine–glutamate transporter) and promoting glutathione synthesis. This enhances cancer development and increases chemotherapy resistance. We investigate the role of CD44v in the regulation of the ROS defense system in cholangiocarcinoma (CCA). Immunohistochemical staining of CD44v and p38^MAPK (a major ROS target) expression in Opisthorchis viverrini‐induced hamster CCA tissues (at 60, 90, 120, and 180 days) reveals a decreased phospho‐p38^MAPK signal, whereas the CD44v signal was increased during bile duct transformation. Patients with CCA showed CD44v overexpression and negative‐phospho‐p38^MAPK patients a significantly shorter survival rate than the low CD44v signal and positive‐phospho‐p38^MAPK patients (P = 0.030). Knockdown of CD44 showed that xCT and glutathione levels were decreased, leading to a high level of ROS. We examined xCT‐targeted CD44v cancer stem cell therapy using sulfasalazine. Glutathione decreased and ROS increased after the treatment, leading to inhibition of cell proliferation and induction of cell death. Thus, the accumulation of CD44v leads to the suppression of p38^MAPK in transforming bile duct cells. The redox status regulation of CCA cells depends on the expression of CD44v to contribute the xCT function and is a link to the poor prognosis of patients. Thus, an xCT inhibitor could inhibit cell growth and activate cell death. This suggests that an xCT‐targeting drug may improve CCA therapy by sensitization to the available drug (e.g. gemcitabine) by blocking the mechanism of the cell's ROS defensive system.     

Effects of self-efficacy, social support and knowledge on adherence to PEFR self-monitoring among adults with asthma: a prospective repeated measures study

BACKGROUND: Long-term adherence to self-monitoring of peak expiratory flow rate (PEFR) is essential for early detection of declining lung function in individuals with asthma. Psychosocial and cognitive factors can influence adherence to PEFR self-monitoring behaviors. OBJECTIVES: The specific aims of this prospective, repeated measures study were to: (1) determine the effects of asthma self-efficacy, perceived satisfaction with social support and asthma knowledge on adherence to PEFR self-monitoring behavior; and (2) examine whether adherence to PEFR self-monitoring mediates the effects of psychosocial/cognitive factors on lung function and asthma symptoms in adults with asthma. DESIGN: Sixty-eight participants completed standardized questionnaires three times at baseline, 1 month, and 3 months and kept the records of PEFR self-monitoring behaviors twice a day. Data were analyzed using multiple linear regressions. Adherence rates to PEFR self-monitoring were 93.5% and 74.9% at 1 and 3 month, representing those who remained in the study (n=39). Other participants withdrew from the study. FINDINGS: Both at 1 and 3 month, psychosocial/cognitive factors as a whole did not account for a significant variance in adherence to PEFR self-monitoring. Univariate analyses, however, indicated that baseline asthma self-efficacy and asthma knowledge at 1 month were significant independent predictors for adherence to PEFR self-monitoring at 3 month. CONCLUSION: Adherence to PEFR self-monitoring did not mediate the effects of asthma self-efficacy, perceived satisfaction with social support, and asthma knowledge on lung function and asthma symptoms. Lung function was low, but participants reported low asthma symptoms, both of which remained stable over time. Because of a small sample size and high attrition, the findings of the study need to be interpreted with caution. Given the importance of long-term adherence to self-management in asthma and other chronic illnesses, factors influencing adherence need to be further investigated to set a basis for future interventions.     

Blastocystis hominis infection in irritable bowel syndrome patients

Irritable bowel syndrome (IBS) is a functional bowel disorder in which abdominal pain is associated with a defect or a change in bowel habits. Subtle inflammation, especially after infectious enteritis, has been sometimes suspected as one mechanism of pathogenesis. This research was performed (1) to evaluate the prevalence of parasitic infections and (2) the possible association of IBS and parasitic infections. Fifty-nine IBS patients were recruited using symptom-based criteria (Rome Criteria II) with an absence of intestinal parasitic infection by direct smear method. Stool samples of individual patients were examined using 7 methods, ie examination for stool occult blood, simple saline smear method, formalin-ether technique, culture for Blastocystis hominis, modified trichrome stain, modified Ziehl-Neelsen method, and trichrome stain for parasitic and bacterial infections. Of the 59 patients, stool samples of 13 patients (22.1%) were positive for parasites. These were B. hominis (13.6%), Strongyloides stercoralis larvae (1.7%), Giardia lamblia cysts (1.7%), and non-pathogenic protozoa, ie Endolimax nana cysts (5.1%). The prevalence rate of parasitic infections in the control group (20%) was not statistically different from the patients. There was no statistical difference between B. hominis infection in IBS patients and control was found in this study (p = 0.87). In the IBS group, B. hominis infection predominated (13.6%), while other parasitic infections were found in 8.5%. The culture method for B. hominis is more sensitive than the direct (simple) stool smear method, which is the routine diagnostic method in most laboratories. These results were also found in control group.     

Methylenetetrahydrofolate reductase (MTHFR) polymorphism (C677T) in relation to homocysteine concentration in overweight and obese Thais

We analyzed the association between MTHFR (C677T) gene polymorphism with serum concentrations of homocysteine, folate, and vitamin B12 in 37 male and 112 female overweight/ obese Thai volunteers (BMI > or = 25.00 kg/m2), and compared them with 23 male and 90 female control subjects (BMI = 18.5-24.99 kg/m2). Statistically significant higher levels of serum homocysteine were found in the overweight/obese subjects than the control subjects (p < 0.05). Serum folic acid levels in the overweight/obese subjects were significantly lower than the control subjects (p < 0.05). When the data were grouped according to homocysteine concentration and MTHFR gene polymorphism, there were significantly higher homocysteine concentrations in the overweight/obese subjects than the control subjects in wild type gene polymorphism (CC) in the hyperhomocysteine group (homocysteine >10.0 mmol/l) (p < 0.05), but in genotype polymorphism (CC, CT, TT) there were lower folic acid and vitamin B12 concentrations in the overweight/obese subjects than in the control subjects. In the hyperhomocysteine groups, there was no significant difference in the frequencies of MTHFR (C677T) gene polymorphism between the overweight/obese subjects and the control subjects. Folic acid and gene polymorphism were found to be significantly related to the overweight/ obese and control groups in logistic regression analysis (p < 0.05). The results support the supposition that folic acid is more important than vitamin B12.     

Type I and type II crustins from Penaeus monodon, genetic variation and antimicrobial activity of the most abundant crustinPm4

An antimicrobial protein, crustin, is involved in the innate immunity of crustacean by defending the host directly against the microbial pathogens. By data mining the Penaeus monodon EST database, two type I crustins, carcininPm1 and 2, and ten type II crustins, crustinPm1–10, were identified. The abundant crustins were crustinPm1, 4 and 7, each with variation in the length of Gly-rich repeat among its members. A few crustinPm1, 4 and 7 with deletion in the Cys-rich region were also observed. Furthermore, the crustinPm4 with the longest N-terminal Gly-rich region was characterized. The crustinPm4 allelic genes were expressed mainly from the hemocytes. Its expression was up-regulated readily by WSSV infection and gradually decreased to normal level afterwards. The recombinant crustinPm4-1 (rcrustinPm4-1) isoform was produced using the Escherichia coli expression system and tested for its antimicrobial activity. The rcrustinPm4-1 was able to inhibit the growth of a Gram-positive bacterium, Bacillus megaterium but not Bacillus subtilis, Micrococcus luteus and Staphylococcus aureus. It also inhibited the growth of two Gram-negative bacteria, E. coli 363 and Vibrio harveyi 639 at lower potency. The rcrustinPm4-1 affected the WSSV infection because the expression of an intermediate early gene ie1 in WSSV-infected hemocyte cell culture was reduced. It was shown further that the rcrustinPm4-1 could delay by about one and a half days the manifestation of disease by WSSV.     

Genes encoding plastid acetyl-CoA carboxylase and 3-phosphoglycerate kinase of the Triticum/Aegilops complex and the evolutionary history of polyploid wheat

The classic wheat evolutionary history is one of adaptive radiation of the diploid Triticum/Aegilops species (A, S, D), genome convergence and divergence of the tetraploid (Triticum turgidum AABB, and Triticum timopheevii AAGG) and hexaploid (Triticum aestivum, AABBDD) species. We analyzed Acc-1 (plastid acetyl-CoA carboxylase) and Pgk-1 (plastid 3-phosphoglycerate kinase) genes to determine phylogenetic relationships among Triticum and Aegilops species of the wheat lineage and to establish the timeline of wheat evolution based on gene sequence comparisons. Triticum urartu was confirmed as the A genome donor of tetraploid and hexaploid wheat. The A genome of polyploid wheat diverged from T. urartu less than half a million years ago (MYA), indicating a relatively recent origin of polyploid wheat. The D genome sequences of T. aestivum and Aegilops tauschii are identical, confirming that T. aestivum arose from hybridization of T. turgidum and Ae. tauschii only 8,000 years ago. The diploid Triticum and Aegilops progenitors of the A, B, D, G, and S genomes all radiated 2.5-4.5 MYA. Our data suggest that the Acc-1 and Pgk-1 loci have different histories in different lineages, indicating genome mosaicity and significant intraspecific differentiation. Some loci of the S genome of Aegilops speltoides and the G genome of T. timophevii are closely related, suggesting the same origin of some parts of their genomes. None of the Aegilops genomes analyzed is a close relative of the B genome, so the diploid progenitor of the B genome remains unknown.     

Large transmission cluster of acute hepatitis C identified among HIV‐positive men who have sex with men in Bangkok,Thailand

A rapidly emerging and highly concentrated hepatitis C virus (HCV) outbreak has recently been observed among both acute and chronic HIV‐positive men who have sex with men (MSM) in Bangkok, Thailand. NS5B regions of the HCV genome were amplified using nested PCR and sequenced. Phylogenetic inference was constructed by Maximum Likelihood methods and clusters were identified with support and genetic distance thresholds of 85% and of 4.5%. Forty‐eight (25 acute HIV and 23 chronic HIV) MSM with incident HCV infection were included in the analysis. HCV genotype (GT) was 85% GT 1a and 15% GT 3a or 3b. Median age at HCV diagnosis was 34 (interquartile range, 28‐41) years. 83.3% (40/48) had history of syphilis infection and 36% (16/44) reported crystal methamphetamine use. Only 2 (4%) reported ever injecting drugs, both crystal methamphetamine. In the phylogenetic clustering analysis, 83% belonged to one of two clusters: one large (75%) and one small (8%) cluster. All clusters were GT 1a. MSM with acute HIV infection were more likely to be in a cluster (92%) than those with chronic infection (74%). HCV screening should be regularly performed for MSM in ART clinics, and offering direct‐acting antiviral agents to all MSM with HCV infection might contain the HCV epidemic from expanding further.     

Azathioprine-induced fatal myelosuppression in systemic lupus erythematosus patient carrying TPMT*3C polymorphism

Azathioprine (AZA) is a commonly used immunosuppressant for systemic lupus erythematosus (SLE). Myelosuppression is a serious adverse reaction due to AZA and its metabolites. Thiopurine S-methyltransferase (TPMT) is the rate-limiting enzyme. Variations of TPMT enzyme activity may be responsible for myelosuppression. However, a correlation between certain mutant alleles of low TPMT enzyme activity and myelotoxicity has also been suggested as a factor. We describe herein a case of AZA-induced severe myelosuppression associated with TPMT*3C heterozygous mutant allele in a SLE patient. The patient presented with pancytopenia, sepsis, typhlitis and disseminated intravascular coagulopathy after a short period of AZA therapy. The patient had low TPMT activity and TPMT*3C genotype. Measurement of TPMT activity and determination of TPMT variant allele may identify patients at risk for AZA-induced myelosuppression.