Identification of novel polymorphism in buffalo stanniocalcin-1 gene and its expression analysis in mammary gland under different stages of lactation

Stanniocalcin-1 (STC1) is secreted by the variety of tissues having a major role in the regulation of calcium ions in the involuting mammary gland. The present work aims to sequence and structural characterization as well as expression profiling of STC1 gene in buffalo. Polymorphism identified in the 3-untranslated region (UTR) was analysed by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) genotyping in riverine and swamp buffaloes. Expression profiling of STC1 was performed in different lactation stages of mammary gland and peripheral blood mononuclear cells to study the impact of 3'-UTR polymorphism on its expression. Different polymorphic sites were detected in the entire coding and noncoding regions of riverine and swamp buffaloes, including two INDELs. An identified polymorphic nucleotide locus A324G, having target sites for two miRNAs, namely bta-miR-2382 and bta-miR-1343, reported in cattle, was genotyped by PCR-RFLP to reveal variable allelic distribution among swamp and riverine buffaloes. Gene expression profiling across buffalo mammary tissues representing different lactation stages showed maximum expression of the STC1 gene in the involuting mammary gland. Ruminants’ specific genetic variation has been observed in STC1 and its implication in buffalo mammary gland involution as well as coregulation of gene expression throughmiRNA binding in the 3'-UTR is suggested.

Comparative Study Between Follicular Unit Extraction (FUE) And Non Cultured Melanocyte Transfer (NCMT) In Stable Vitiligo Patients

Background: Vitiligo is an acquired, hypomelanotic disorder characterized by circumscribed depigmented macules in the skin resulting from the loss of functional melanocytes from the cutaneous epidermis. It also causes significant psychological and social distress. Aims and Objectives: To compare the efficacy of follicular unit extraction and non cultured melanocyte transfer in patients of stable vitiligo with respect to repigmentation, vitiligo noticeability and global treatment success. Material and Methods: A total of 15 patients with stable vitiligo (as per IADVL guidelines) were enrolled in the study. In the same patient follicular unit extraction (FUE) was done in the vitiliginous lesions and the hair was transplanted approximately 3-5 mm apart on the left side of the body, while another vitiliginous lesion in the same patient was selected for non cultured melanocyte transfer (NCMT) which was done on the dermabraded area on the right side of the body. These patients were followed-up for a period of 6 months, initially at every 2 weeks or till first signs of repigmentation, then monthly follow-ups for two times and then followed-up in every 2 months. Visual analogue scale was used for assessment of repigmentation, VNS scale was used to evaluate vitiligo noticeability and global treatment success was calculated. Results: There were 2 (13.3%) females and 13 (86.7%) males in our study, showing a male preponderance. Majority of the patients were in the age group 21-40 years (66.7%). There was statistically significant increase in the mean pigmentation at each follow-up in comparison to the earlier follow-up in both the groups (p<0.05). The mean pigmentation and mean pigmentation difference, between the two groups was also comparable (p>0.05). Excellent pigmentation was seen in 60% patients of FUE and 73.3% patients of the NCMT group. Vitiligo was ‘not noticeable’in 33.3% patients of FUE and 40.0% patients of NCMT group. Global treatment success was 80% in both the groups. Bony prominence, greying and loss of follicles in FUE group; and graft displacement and herpes zoster in NCMTgroup were the factors responsible for low pigmentation. Conclusion: From the above study, we conclude that repigmentation was seen in both the groups, with equal efficacy seen between the two methodologies. Thus, any method can be applied for repigmentation with due considerations to complications of each method used.

Anti - microbial resistance stratified by risk factor among Escherichia coli strains isolated from the urinary tract at a rural clinic in Central India.

Background: The failure of empirical therapy is frequently observed, even in community-acquired urinary tract infections. We, therefore, conducted a prospective, clinic-based study in 2004-2005 to document anti-microbial resistance rates and correlate them with possible risk factors to assist empirical decision-making. Materials and Methods: Symptomatic patients with pyuria underwent urine culture. Isolates were identified using standard methods and anti-microbial resistance was determined by disk-diffusion. Ultrasonography was used to detect complicating factors. Patients were stratified by the presence of complicating factors and history of invasive procedures for comparison of resistance rates. Statistical Method Used: Chi-square or Fisher exact tests, as appropriate. Results: There were 156 E. coli isolates, of which 105 were community-acquired. Twenty-three community-acquired isolates were from patients with complicating factors while 82 were from patients without any. Fifty-one isolates were from patients who had recently undergone invasive procedures on the urinary tract. Thirty-two community-acquired isolates from reproductive-age women without apparent complicating factors had resistance rates of 50% or above against tetracyclines, Co-trimoxazole, aminopenicillins, Nalidixic acid, Ciprofloxacin and 1 st generation cephalosporins. Resistance rates were significantly higher among isolates from patients subjected to invasive procedures, except against Co-trimoxazole, tetracyclines and Amikacin. Conclusion: High rates of anti-microbial resistance in community-acquired uropathogens have made antimicrobial sensitivity testing necessary even in a rural, primary-care setting.

Characterisation of fowl adenoviruses from chickens affected with infectious hydropericardium during 1994-1998 in India.

In the present study characterisation has been done for six group I fowl adenoviruses (FAV) isolated from outbreaks of infectious hydropericardium (IHP) of chickens that occurred in different states/regions of India during the years 1994-98. These six viruses were identified as FAV serotype 4 by virus neutralisation and restriction endonuclease analyses. Antigenic analyses of the viruses revealed close relationship (R-values 0.93-0.96). Under the experimental conditions, we have been able to induce IHP using FAV serotype 4 isolate AD: 411 and were also able detect FAV antigens in myocardial tissues by immunofluorescence assay (a new observation), an indication that IHP causing FAV serotype 4 strain replicate in myocardial tissue. Restriction endonuclease analysis of the viral genomes (approximately 46 Kb), using Hind III, Sma I, Xba I, Bam HI, Pst I and Dra I produced identical genetic profiles. Pst I and Bam HI profiles for these six vitus isolates were identical to those published earlier for an IHP causing Pakistani FAV serotype 4 isolate KR31. The identical genetic profiles of viruses, chronology of the outbreaks of IHP in Pakistan during 1989 onward and later in Jammu and Kashmir, India (1994), suggest that FAV serotype 4 isolates involved in outbreaks of IHP in India had probably spread from Pakistan. In order to prevent further spread and economic losses due to IHP in India, based on the antigenic relatedness data in this paper, any one of the six studied FAV serotype 4 isolates can be used as a candidate for mass production of CEH culture based killed vaccine.

Maxillary mass as the presenting manifestation of papillary thyroid carcinoma.

A case of follicular variant of papillary thyroid carcinoma presenting with a right maxillary mass is described. This is perhaps the first instance of maxillary metastasis from papillary thyroid carcinoma.

Parathyroid carcinoma: difficult management options.

Parathyroid carcinoma is a rare cause of primary hyperparathyroidism and these tumours are usually hyperfunctional as opposed to other malignant endocrine tumors. Surgery is the only effective treatment while nonsurgical modalities yield poor results. We report a patient, who presented with palpable mass in the neck and severe hypercalcemia. He underwent debulking surgery and received allendronate, calcitonin, dacarbazine followed by in- situ alcohol instillation with some success.

Purification and differentiation of three avian adenoviruses by restriction endonuclease analysis.

A rapid method of ultracentrifugation pelleting of avian adenovirus (AAV) from small volume of chloroform treated infected cell culture fluid or allantoic fluid was adapted for isolation of adenoviral DNA. The viral DNA extracted from semipurified viruses was found to be intact on agarose gel and pure enough (A260/280 = 1.85-1.92) for restriction enzyme analysis. Restriction endonuclease analysis of Indian strain of AAV serotype 1, AAV serotype 4 (group I AAVs) and egg drop syndrome-76 (EDS-76) virus genomes (group III AAV) with Hind III enzyme differentiated these viruses. The AAV serotype 1 and serotype 4 strain exhibited identical Hind III profile to European viral strains belonging to same serotypes however, the EDS-76 virus gave similar but not identical profile. The calculated genomic lengths for AAV serotype 1 and EDS-76 virus were approximately found to be 33.9 and 44.4 Kb, respectively.

Spectrum of cystic variants of Wilm's tumour: cystic nephroma (multilocular cyst) and cystic partially differentiated nephroma--a report of four cases.

Two cases of cystic nephroma (multilocular cyst of the kidney), and one case each of cystic partially differentiated nephroblastoma (CPDN) and rhabdomyomatous Wilms' tumour are described. All were male and in the pediatric age group. Grossly tumours were unilateral, unicentric and multiloculated. The need for proper designation of these lesions is highlighted because of difference in the treatment and prognosis of these tumours.

Gastrointestinal mucormycosis--an uncommon isolated mucormycosis.

BACKGROUND: Isolated mucormycosis of the gastrointestinal tract is uncommon, with only two case reports from India. OBJECTIVES: To study the clinicopathologic features of gastrointestinal mucormycosis in Indian patients. METHODS: Eight cases of isolated gastrointestinal mucormycosis, seen over six years (1992-97) are reviewed. RESULTS: Five of the patients were premature babies or infants, one was a 12-year-old boy, and two were middle aged. Symptomatology included vomiting, bloody diarrhea, upper gastrointestinal bleeding, abdominal lump and abdominal distention. All the neonates presented with intestinal perforation. The duration of symptoms was ten days or less in six cases. Mucormycosis was not suspected clinically in any patient. In six cases the diagnosis was established antemortem from resection or biopsy material. Only two patients received antifungal therapy and only one patient responded. CONCLUSIONS: Isolated gastrointestinal mucormycosis is not uncommon in India. Early diagnosis may be helpful in reducing the high mortality.

Detection of bluetongue virus genome segment 6 sequences by RT-PCR.

Reverse transcription-PCR (RT-PCR) technique was adopted to amplify a 101 basepair nucleotide sequence of bluetongue virus (BTV) genome segment 6. The specificity of the amplicon was determined by its approximate size in 3% agarose gel electrophoresis, digestion with restriction enzyme MspI, dot-blot hybridization and cycle sequencing. The technique was found to be suitable for detection of bluetongue virus in infected cell culture and clinical samples.

Clinical course and management of pancreatoblastoma in children.

Pancreatoblastoma is a rare malignant tumour. Two children with this tumour were managed in the last 2 years. Both presented with progressively increasing abdominal mass. The diagnosis was established only after laparotomy. In the first child, an 8 year old girl, the mass was arising from the body of the pancreas and only incomplete resection was feasible. She received postoperative chemotherapy and went into remission for a few months before presenting with jaundice and abdominal pain due to recurrent, metastatic disease in the liver and porta hepatitis. Further therapy was refused by the patient because of anorexia and social problems. The second patient, a 5-year-old girl, underwent distal pancreatectomy for complete removal of a large mass arising from the tail of the pancreas. Chemotherapy was begun postoperatively but discontinued by the patient. However, she has remained disease free 1 year after diagnosis. Histologic, histochemical and ultrastructural features of the tumour are detailed and the management discussed.

Helicobacter pylori infection in patients with perforated duodenal ulcer.

AIM: Perforation is the commonest complication of duodenal ulcer. Helicobacter pylori is found in 95% patients with duodenal ulcer. However, there is paucity of reports on prevalence of H. pylori infection in patients with duodenal ulcer perforation. We, therefore compared the incidence of H. pylori infection in patients with duodenal ulcer perforation with the incidence in patients having complicated duodenal ulcers and non-ulcer dyspepsia. PATIENTS AND METHODS: The study was conducted on 45 patients (complicated duodenal ulcer 15, duodenal ulcer perforation 15, non-ulcer dyspepsia 15). Per-operative punch antral biopsies were taken in patients with duodenal ulcer perforation whereas endoscopic punch biopsies of antrum were taken in patients with non-ulcer dyspepsia. The criteria for H. pylori positivity was i) growth of H. pylori on culture, ii) combination of rapid urease test (RUT) and Giemsa staining, combination of RUT and Gram stain being positive for H. pylori. RESULTS: While 9 of 15 cases with complicated duodenal ulcer, 7 of 15 cases with non-ulcer dyspepsia were positive for H. pylori, none of the patients with duodenal ulcer perforation tested positive for H. pylori (p < 0.000). All patients with perforated duodenal ulcer had histological gastritis (H. pylori -ve). Fourteen of 15 patients (9 H. pylori +ve, 5 H. pylori -ve) with complicated duodenal ulcer and 9 of 15 patients (7 H. pylori +ve) with non-ulcer dyspepsia had histological gastritis. CONCLUSION: Patients with duodenal ulcer perforation do not have H. pylori infection. H. pylori negative patients of duodenal ulcer may have more predilection for perforation.

Neurogenic bladder: urodynamic and surgical aspects.

Children with spina bifida often have vesico-urethral dysfunction manifesting either as upper urinary tract deterioration or voiding dysfunction, chiefly incontinence. Surgery of the back and presence of or increase in hydrocephalus may contribute to neuro-urologic worsening; secondary cord tethering and syrinx or hydromyelia may be additional factors coming into play later in life. Urodynamic assessment using simple modalities like uroflowmetry, external sphincter EMG, residual urine volume and cystometry provide data useful to classify patients according to detrusor and sphincter activity. Besides diagnosis, urodynamic studies are useful in guiding therapy of children with vesico-urethral dysfunction, and for their follow-up to detect sub-clinical deterioration. They also help to prognosticate risk of upper tract deterioration and the possible success of measures to contain incontinence. Urodynamic data in thirty one patients with spinal dysraphism who presented to us with urologic symptoms were analysed. Twenty-three children had hyper-reflexic bladders while in the other 8 the bladder was areflexic. 13 children showed upper tract dilatation. The leak point volume was significantly lower in this group of patients compared to those who did not show upper tract dilatation. Our results are comparable to earlier similar studies.

Occult spinal dysraphism.

When spina bifida is associated with skin abnormalities such as dimples, sinus tracts hypertrichosis, or capillary hemangiomas, there is a high incidence of an occult intraspinal lesion such as epidermoid and dermoid tumours, lipomas, diastematomyelia, dural bands and tethered spinal cord. The present study consists of 50 patients with occult spinal dysraphism treated with the diagnosis of lipomeningomyelocoele (20), lumbosacral lipoma (15) and diastematomyelia (15). The clinical presentation varied from lipoma in the lumbosacral region, dermal sinus, cutaneous hemangioma and hypertrichosis. The age range varied from 2 months to 7 years with an average of 2 years. 40% patients had neurological deficit at the time of presentation which varied from lower limb weakness to bladder & bowel involvement. All patients underwent laminectomy of the lumbar and lumbo-sacral spine with excision of intraspinal lipoma, excision of bony or cartilaginous spur in diastematomyelia and detethering of the conus medullaris & cauda equina. No patient developed late neurological deficit.

Management of hydrocephalus.

The present study is an analysis of 747 patients with hydrocephalus, treated and followed up in the Hydrocephalus Clinic run by the department of Paediatric Surgery at the All India Institute of Medical Sciences, New Delhi. The distribution of patients was: congenital-46%, post-meningomyelocoele excision-28%, post-meningitic-21% and others-5% (including post haemorrhagic and post encephalocoele excision hydrocephalus. The average age was 7 months in the shunted group and 10 months in the medical group with overall male to female ratio of 2.3:1. The data were analysed to study the effect of treatment on ventriculomegaly and mental development with special reference to the type of treatment (shunt versus medical) and age at starting treatment. The probability of shunt failure was also studied. A comparison of ventricular size in US/CAT scans between the time of starting treatment and last follow-up revealed improvement in ventriculomegaly in 60% of the shunted patients but only 30% of the medically treated patients. A significant difference was particularly noted in patients with severe hydrocephalus, 72% and 22%, respectively. Comparison of the mean Mental Performance Quotient (MPQ) scores in the shunted & medically treated patients also revealed significantly better MPQ scores in the shunted group (p = < 0.001). Probability of shunt survival, as depicted by the Kaplan-Meier survival curve, revealed that there is a high rate of shunt failure in the first 12 months, followed by a dramatic slowing down. Our observations support the contention that CSF shunt surgery offers better outcome than medical management even when ventriculomegaly is severe at the time of presentation.

Solid cystic tumour of the pancreas-clinico-pathological study of three cases.

During a period of four years three cases of solid and cystic (papillary) tumour of the pancreas were encountered. The patients presented predominantly with chief complaints of lump abdomen associated with dull aching pain in right hypochondrium. Radiologically diagnosis was suspected in two cases. Excessive bleeding during surgery was observed in two cases. In two survivors no recurrence or distant metastasis was noted after surgical excision (follow up 6 months to 3.5 years), third patient died after eight hours of operation.

Role of extramucosal seromuscular biopsy during preliminary fecal diversion in the management of Hirschsprung's disease.

The routine use of frozen section biopsy to identify distal ganglionic bowel in Hirschsprung's disease is the accepted norm in well developed centers. With the aim of overcoming non-availability of frozen section biopsy and reducing the duration of definitive surgery, an alternative algorithm, based on extramucosal seromuscular biopsy at the time of preliminary fecal diversion and its examination by routine paraffin sections and staining, was developed and used in 100 patients at our centre. This technique accurately marked the ganglionic colon for pull-through operation in 96.2% of cases with rectosigmoid transition zone and in 91.7% of those with transition zone in the descending colon, thus obviating the need for frozen section biopsy and saving nearly one hour during the pull-through surgery. However, in the more proximal varieties of Hirschsprung's disease frozen section biopsy was still considered to be indispensable.

Spectrum of congenital anomalies associated with biliary atresia.

Several congenital anomalies have been reported in association with biliary atresia. We have analysed the type and frequency of anomalies observed over a 10-year period in consecutive patients operated for extrahepatic biliary atresia at our institution. Of the 107 infants who underwent a laparotomy and surgical correction of biliary atresia, 9 (8.4%) showed significant associated anomalies. Among them, 5 (55.5%) had splenic malformations, 6 (66.6%) had digestive anomalies in the form of malrotation, Meckel's diverticulum and jejunal atresia. One patient had the classical polysplenia syndrome. Our follow-up period was limited (i.e. 5 months) during which time 3 of the 9 (33.3%) patients with associated anomalies became jaundice-free. A higher incidence of malformations found in association with biliary atresia supports the congenital theory and a more thorough search for these anomalies is recommended.