Gene Expression Profiling and Assessment of Vitamin D and Serotonin Pathway Variations in Patients With Irritable Bowel Syndrome

BACKGROUND/AIMS: Irritable bowel syndrome (IBS) is a multifaceted disorder that afflicts millions of individuals worldwide. IBS is currently diagnosed based on the presence/duration of symptoms and systematic exclusion of other conditions. A more direct manner to identify IBS is needed to reduce healthcare costs and the time required for accurate diagnosis. The overarching objective of this work is to identify gene expression-based biological signatures and biomarkers of IBS. METHODS: Gene transcripts from 24 tissue biopsy samples were hybridized to microarrays for gene expression profiling. A combination of multiple statistical analyses was utilized to narrow the raw microarray data to the top 200 differentially expressed genes between IBS versus control subjects. In addition, quantitative polymerase chain reaction was employed for validation of the DNA microarray data. Gene ontology/pathway enrichment analysis was performed to investigate gene expression patterns in biochemical pathways. Finally, since vitamin D has been shown to modulate serotonin production in some models, the relationship between serum vitamin D and IBS was investigated via 25-hydroxyvitamin D (25[OH]D) chemiluminescence immunoassay. RESULTS: A total of 858 genetic features were identified with differential expression levels between IBS and asymptomatic populations. Gene ontology enrichment analysis revealed the serotonergic pathway as most prevalent among the differentially expressed genes. Further analysis via real-time polymerase chain reaction suggested that IBS patient-derived RNA exhibited lower levels of tryptophan hydroxylase-1 expression, the enzyme that catalyzes the rate-limiting step in serotonin biosynthesis. Finally, mean values for 25(OH)D were lower in IBS patients relative to non-IBS controls. CONCLUSIONS: Values for serum 25(OH)D concentrations exhibited a trend towards lower vitamin D levels within the IBS cohort. In addition, the expression of select IBS genetic biomarkers, including tryptophan hydroxylase 1, was modulated by vitamin D. Strikingly, the direction of gene regulation elicited by vitamin D in colonic cells is “opposite” to the gene expression profile observed in IBS patients, suggesting that vitamin D may help “reverse” the pathological direction of biomarker gene expression in IBS. Thus, our results intimate that IBS pathogenesis and pathophysiology may involve dysregulated serotonin production and/or vitamin D insufficiency.

Antibacterial activity of Elaeagnus umbellata [Thunb.] a medicinal plant from Pakistan

To evaluate the biological activity of Elaeagnus umbellata extracts on standard microorganism strains as well as multi-drug resistant bacteria isolated from hospitals. We carried out this study at the Plant Pathology Laboratory of the University College of Agriculture, Rawalakot Pakistan during the period between September-November 2004. Flowers, leaves, and berries of the plant were extracted in different solvents and tested for their antibacterial activity by disc diffusion method on selected organisms like methicillin resistant Staphylococcus aureus [S. aureus], multi-drug resistant Pseudomonas aeruginosa [P. aeruginosa], and enterohemorrhagic Escherichia coli [E. coli]. The ether extract of flower was found to be highly effective against E. coli, P. aeruginosa, S. aureus, and Bacillus subtilis [B. subtilis]. The alcohol extract of these leaves also demonstrated strong activity against gram positive and negative bacteria. The aqueous extract from the berry strongly inhibited the growth of E. coli and S. aureus whereas, it exhibited a very small zone of inhibition against B. subtilis. Multi-drug resistant P. aeruginosa was found completely resistant to aqueous extract. The acetone extract of the berry showed good activity against P. aeruginosa. The present study reports the antibacterial activity of Elaeagnus umbellata. Most of the extracts displayed broad-spectrum activity, since gram positive bacteria including S. aureus, B. subtilis and gram-negative bacteria including E. coli and P. aeruginosa were inhibited. These preliminary findings may provide the basis for traditional use of this plant in the treatment of infectious diseases

Hepatitis B and C infection in first- time blood donors in Karachi - a possible subgroup for sentinel surveillance

Hepatitis B virus [HBV] and C virus [HCV] are major public health concerns in Pakistan. We conducted a baseline analysis of first- time replacement blood donors at the blood bank of Jinnah Postgraduate Medical Centre in the year 2000 with the view to developing low- cost sentinel surveillance for these infections. Among 7325 such donors, 264 [3.6%] were positive for anti- HCV and 344 [4.7%] for HBsAg. HCV seroprevalence was significantly positively associated with age and lower education. Those speaking Sindhi or other minor languages had a significantly higher prevalence of HBV infection. Blood bank data could provide reliable information to monitor trends in prevalence of these infections

Use of intravenous anti-D in patients with refractory and relapsed immune throm bocytopenic purpura

To determine the response to IV anti-D and its comparison with splenectomy as second line therapy in refractory and relapsed cases of ITP, in the Aga Khan University Hospital, Karachi. A total of 23 patients with chronic ITP were treated with either anti-D or splenectomy as second line treatment. The patients were assessed for time to achieve a response to second line treatment, duration of response and adverse events. There were 12 patients in the anti-D group and 11 in the splenectomy group. The mean platelet count at presentation was 9,000/cumm. The mean age was 8.9 years and 13.0 years and the male to female ratio was 1:1 and 1:1.2 in anti-D and splenectomy group respectively. 54.5% of the patient in the anti-D group responded compared to 81.8% in the splenectomy group. Median time to achieve a response was 7 days in the anti-D group and 1 day in the splenectomy group. Mean time to relapse was 87.8 days in the anti-D group and 55.4 days in the splenectomy group. No adverse events were recorded for any of the infusions of anti-D and none of the patients had more than 0.5 gm /dl fall in the hemoglobin level following anti-D infusion. It was thus concluded that Anti-D is a relatively safe, convenient and effective therapy for chronic ITP and can be used as a splenectomy sparing agent when treatment is clinically indicated

Infective morbidity following cesarean section

To find out the incidence and causes of infection following Caesarean section, to study risk factors and to prove the effectiveness of preoperative antibiotic prophylaxis in prevention of postoperative infections. Design: A prospective study of all pregnant cases who had Caesarean section during November, 1995 to April 1996. Setting: Dept. of Obstetrics and Gynaecology, Unit-1, Sir Ganga Ram Hospital, Lahore. Subjects: A total of 494 patients were managed in postnatal ward during the study period. Main out come Measures: To recognize avoidable risk factors and to find out ways and means to reduce infective morbidity resulting from Caesarean section. During the study period total number of Caesarean section performed were 494. Emergency C. Section were done in 425 patients where an elective C. section was performed in 69 patients. Post-operative pyrexia persisting after first 24 hours was investigated and treated. Postoperative infection rate was 14.37%. Majority of the patients suffered from urinary tract infection [35.21%] followed by endometritis [33.81%]. Wound infection and disruption was found in 9.86%. The effectiveness of prophylactic antibiotics was analysed. Vigilant intrapartum monitoring, surgical skill and antibiotic prophylaxis can reduce the incidence of postoperative infective morbidity