Characterisation of the growth and differentiation in vivo and in vitro-of bloodstream-form Trypanosoma brucei strain TREU 927.

Trypanosoma brucei TREU 927/4 has been chosen as the reference strain targeted for complete sequencing of the genome of the African trypanosome. This line is pleomorphic in mammalian hosts and is fly transmissible; however it is relatively unstable with respect to variable surface glycoprotein (VSG) expression. Therefore, we subjected TREU 927/4 to 27 rapid syringe passages through mice, and derived a cloned line which expressed Glasgow University Trypanozoon antigen type (GUTat) 10.1 with relative stability. This line also retained pleomorphism in the bloodstream, being able to generate homogeneous populations of stumpy forms in mice. Furthermore, these parasites remain able to transform to procyclic forms synchronously in vitro and can complete their life cycle in tsetse flies. The passaged cell line was also adapted to in vitro bloodstream-form culture and transfected with a construct encoding the tetracycline repressor (TETR) protein. The resulting TETR subline no longer expressed the GUTat 10.1 VSG but remained able to generate uniform populations of stumpy form cells in mice immunocompromised with cyclophosphamide. They could also differentiate to procyclic forms synchronously in vitro. The generated lines and analyses of their growth and differentiation will provide a basic resource for the analysis and interpretation of gene function in the T. brucei genome reference strain.

Detection and functional characterisation of the transcription factor peroxisome proliferator-activated receptor gamma in lutein cells.

A prominent functional change during differentiation of lutein cells from follicular thecal and granulosa cells is an enhanced production and secretion of progestins. The regulation of this process is not fully understood but may be associated with the expression of transcription factors which activate genes, products of which are involved in pathways of the cholesterol and lipid metabolism. As peroxisome proliferator-activated receptors (PPARs) play a role in both pathways, we were interested in the expression of PPARgamma, a PPAR form which is involved in adipogenic differentiation. First, we were able to show the expression of PPARgamma in bovine lutein cells (day 12 of the ovarian cycle) at the mRNA and protein level by imaging, flow cytometry and blot analysis, and secondly a role of PPARgamma in the secretion of progesterone. The cells (24 h culture) responded dose dependently by increasing progesterone secretion (up to 1.5-fold of the basal level) to an endogenous ligand of PPARgamma, 15-deoxy-delta12,14 prostaglandin J2 (15-dPGJ2) and to the thiazolidinedione ciglitizone. Aurintricarboxylic acid (ATA) was found to reduce the intracellular PPARgamma level and to promote cell cycle progress, indicating that ATA can be used as a tool for experimental changes of PPARgamma proteins in intact cells and for studying the physiological consequences. The ATA-mediated decrease of PPARgamma was accompanied by reduced progesterone production and a progression of the cell cycle, suggesting a function of PPARgamma in both processes. The response to ATA was abrogated by a high dose (>490 nM) of 15-dPGJ2, suggesting that 15-dPGJ2 exerts its effect on steroidogenic activity via PPARgamma and that the 15-dPGJ2-PPARgamma system plays a role in the maintenance of a differentiated quiescent stage in lutein cells.

Triple-drug therapy of Helicobacter pylori infection in duodenal ulcer disease.

OBJECTIVE: To evaluate the efficacy of triple-drug therapy for Helicobacter pylori infection and the effect of its eradication on relapse of ulcer in patients with duodenal ulcer (DU) disease. METHODS: Patients with uncomplicated DU who were H. pylori-positive on urease test or histology were given triple-drug therapy (metronidazole, tetracycline, colloidal bismuth subcitrate). Ulcer healing and H. pylori status were assessed one month after completion of therapy. Those with healed ulcers were followed up endoscopically for ulcer recurrence at 3-month intervals for one year or more. RESULTS: Fifty seven of 60 consecutive DU patients (95%) who were H. pylori-positive were taken up for the study; 46 patients (36 men; median age 40 years, range 13-70) completed the study. Thirty one of them (67%) eradicated H. pylori and had healed ulcers at one month post therapy; of 15 patients with persistent H. pylori infection, ulcers healed in 12 (p = ns). After an average follow up of 11 months, 8 of 12 H. pylori-positive patients had relapse of DU compared to 5 of 31 (16%) H. pylori-negative patients (p < 0.01). CONCLUSIONS: Triple-drug therapy was effective and safe for H. pylori infection, the lower rate of eradication observed compared to results elsewhere probably being related to bacterial resistance. Eradication of H. pylori was associated with healing of ulcers, resolution of antral gastritis and a significant fall in the rate of ulcer relapse.

A study of keratomycosis: our experience.

The aim of the present study was to determine the etiological agents, predisposing factors and important diagnostic tests in keratomycosis in Patna. During period of 2 years, 204 clinically suspected cases of keratomycosis were studied by microscopy (10% KOH smear preparation and Gram's staining) and culture (for bacteria as well as fungus). Out of 204 clinically suspected cases, 76 cases (37.23%) were reported positive for fungus by microscopy and culture. 77% cases were between the age group of 21 to 50 years. Males were more than double the number of females affected. 82% patients were farmers by occupation. 81.58% cases gave a definite history of antecedent corneal trauma. 6.58% cases were known diabetics. Peak incidence was recorded in the months of September-October of the years. Single fungal isolates were present in 63 cases (31%) and multiple were there in 3 cases (1.47%). Mixed isolates of bacteria and fungi were there in 4.90% cases. The predominant isolate was Aspergillus species--42 cases (52.26%) followed by Candida--7 cases (9.21%), Penicillium--6 cases (7.89%) and Fusarium-6 cases (7.89%). Ten (13.15%) of fungal isolates remained unidentified. Therefore mycotic keratitis should be suspected in every patient with a corneal lesion and should be ruled out promptly to save sight and before commencing steroids and antibiotics.

Serum magnesium level in preterm labour.

Preterm labour, (PTL) defined as labour after 28 weeks but before 37 completed week of gestation, is an ill omen for our country as the incidence is 5-10% leading to 70-80% of perinatal deaths. Besides varied etiology, it may be due to alteration in basic biochemical functions of the body at cellular level stating emphasis to trace elements, of which magnesium, being one of them, is subject of interest now a days. Serum magnesium was estimated in fifty cases of Preterm labour of idiopathic etiology (28-37 wks), 35 normal pregnancy cases of the same gestational age and 15 normal non-pregnant ladies. Varied hypomagnesemia was observed in Preterm labour cases (1.47 mg/dl +/- 0.22 S.D.), normal value of serum magnesium was found in normal non-pregnant ladies and slightly low value were observed in pregnant ladies of same gestational age. Age and parity had no significant effect on serum magnesium level in our study. As far as socio-economic study is concerned, it was found to be higher in high socio-economic group and low in lower group. Thus from this study it can be concluded that estimation of serum magnesium in pregnancy may prove to be a valuable tool in predicting preterm onset of labour.

Covert bacteriuria in nephrotic syndrome.

Prevalence of covert bacteriuria in patients of nephrotic syndrome admitted for kidney biopsy was studied in 205 patients. Age ranged from 10 years to 65 years. 148 patients were male and 57 were female. Prevalence of covert bacteriuria was found in 38 patients (18.53%). In bacteriuric patients 30 were male, 8 were female. Escherichia coli was the commonest organism grown in bacteriuric patients (30 or 78.9%). Serum albumin was low and 24-hour urinary protein excretion was high in bacteriuric patients in comparison to abacteriuric patients. In bacteriuric patients membranous nephropathy was the commonest histopathological finding present in 15 patients (39.47%).

Prevalence of hepatitis B and hepatitis C virus coinfection in chronic liver disease.

Coinfection with HBV and HCV may lead to serious consequences. The present study was done to find out the prevalence of coinfection in patients with chronic liver disease. From patients with hepatitis and chronic liver disease 1673 samples were received and analysed for HBsAg by ELISA. 1342 samples were analysed for anti HCV by third generation ELISA. 493 samples positive for HBsAg were also analysed for Anti HCV to see the prevalence of coinfection. 15(3.0%) were found positive for both HBsAg and anti HCV. Out of 15 patients with coinfection 4 (26.6%) had HCC. Prevalence of HCC in patients with coinfection was higher than either infection alone i.e. HBV-9.1% and HCV-16.5%.

Prostate specific antigen as tumor marker: relationship with histologic grading.

PSA is emerging as the best marker in oncology and had a profound impact on all aspects of prostate cancer care. From clinically suspected prostate tumor, 395 serum samples were taken out and estimated for serum PSA. Among elevated serum PSA, 98 were correlated with histologic findings. 42(42.8%) cases were BHP among 98 cases and 78.7% had serum PSA level within 10 ng/ml. 5 patients (5.1%) had PIN histologically, 3(60%) of which had PSA level upto 10 ng/ml and 2(40%) had serum PSA upto 20 ng/ml. 51(52%) were adenocarcinoma prostate of different grades and PSA level varies from less than 10 ng/ml to more than 50 ng/ml which almost correlates with the tumor grades.

Covert bacteriuria in nephrotic syndrome.

Prevalence of covert bacteriuria in patients of nephrotic syndrome admitted for kidney biopsy was studied in 205 patients. Age ranged from 10 years to 65 years. 148 patients were male and 57 were female. Prevalence of covert bacteriuria was found in 38 patients (18.53%). In bacteriuric patients 30 were male, 8 were female. Escherichia coli was the commonest organism grown in bacteriuric patients (30 or 78.9%). Serum albumin was low and 24-hour urinary protein excretion was high in bacteriuric patients in comparison to abacteriuric patients. In bacteriuric patients membranous nephropathy was the commonest histopathological finding present in 15 patients (39.47%).

Coagulase negative Staphylococci as causative agents of urinary tract infections-prevalence and resistance status in IGIMS, Patna.

Previously considered solely as the laboratory contaminants and normal flora of skin in man, coagulase negative Staphylococci are now a major cause of nosocomial and opportunistic infections. This study was conducted at IGIMS, Patna to find out the coagulase negative Staphylococcus isolates from urine and their antimicrobial resistance. In a period of ten months, the relative frequency of main coagulase negative Staphylococci were as follows--Staphylococcus epidermidis-45.90%, Staphylococcus saprophyticus 34% and Staphylococcus haemolyticus-8.50%. others were 11.60%. most of the Staphylococcus saprophyticus was isolated from young female patients suffering from uncomplicated acute cystitis and Staphylococcus epidermidis was mainly from patients with indwelling catheters and complicated cases. Staphylococcus saprophyticus showed the highest sensitivities to all the antimicrobials whereas Staphylococcus haemolyticus had the highest resistance rates. 66.6% of staphylococcus epidermidis and 60% of staphylococcus haemolyticus were resistant to oxacillin, whereas only 10% of staphylococcus saprophyticus resistant and 90% were sensitive to it. As is in the emerging state, vancomycin resistance was very less but in future it may cause a major problem to treat these cases. So this area needs further exploration.

Study on serum sperm agglutination in cases of unexplained infertility.

In our study we investigated 100 couples of unexplained infertility in order to detect the presence of anti-spermatozoal antibodies. Both auto-immunity and Iso-immunity have been found responsible in 12.5% of couples of primary infertility & 10% of couples of secondary infertility. As many as, 21% of cases of unexplained infertility were attributed to presence of antibodies in sera of infertile couples. Here, we highlight the importance of anti-spermatozoal antibodies testing in the first instance itself in cases of unexplained infertility.

Role of enzyme linked immunosorbent assay in the diagnosis of suspected cases of genito urinary tuberculosis.

The aim of study was evaluation of the utility of ELISA test using A60 Antigen for rapid diagnosis of Genitourinary Tuberculosis in various age groups. ELISA test based on mycobacterial antigen A60 (Anda biological, France) was used to estimate specific IgG antibodies in the sera of fifty four suspected cases of Genito urinary tuberculosis. (GUT)Sera of 30 montoux negative healthy adults (age/sex matched) were taken as control by detecting IgG anti bodies to A60 antigen. It was concluded from this study that IgM was positive in 87.0% of cases.

Fasciolopsis buski (giant intestinal fluke)--a case report.

A girl, aged 20 years presented with diarrhoea, vomiting, pain abdomen and loss of weight, the routine Stool examination revealed Fasciolopsis buski (giant intestinal fluke) in large numbers. Despite treatment with Praziquantel, she died after three days.