Cytological and immunocytological detection and differentiation of Marek's disease and lymphoid leucosis in poultry.

Marek's disease (MD) and lymphoid leucosis (LL) are the major diseases causing lymphoid tumors in chickens accounting for high economical losses. Gross examination could not yield definite diagnosis owing to their similar presentation of lesions. Thus present work was aimed for diagnosis and differentiation of MD and LL by utilizing simple cytology and novel immunocytology techniques. Cytological examination was carried out on slides with tumor touch imprints stained by simple Giemsa staining. The diagnosis was mainly achieved based on morphology of cell population. In the present study, out of a total of 595 cases examined, 502 cases had pleomorphic lymphocytic cell population suggestive of MD and 53 cases had uniform lymphocytic/lymphoblast cell population suggestive of LL, while the rest 40 cases remained inconclusive. A definitive diagnosis was achieved after performing immunocytology using specific antibodies that revealed 518 cases had reactivity for Meq oncoprotein specific for MD and 77 cases showed immunoreactivity for IgM in transformed B-cells confirming LL. The technique of immunocytology which has been useful for detecting human viral pathogens and MD in poultry has been applied for the first time as a novel, simple, rapid and inexpensive technique that could be used as an alternate test to effectively detect and differentiate MD and LL in poultry.

A novel recombinant Meq protein based dot-ELISA for rapid and confirmatory diagnosis of Marek's disease induced lymphoma in poultry.

Marek's disease (MD), is an economically important virus disease of poultry throughout the world. In this study, we for the first time reports development of a novel dot enzyme-linked immunosorbent assay (dot-ELISA) for the confirmatory diagnosis of lymphoma caused by Marek's Disease Virus (MDV). Suspected lymphoma tissue extracts from the diseased birds were used for the Meq oncoprotein antigen detection, which is expressed specifically in MDV lymphomas. Recombinant Meq oncoprotein was expressed using Expresso™ Rhamnose Sumo Cloning and Expression system and the hyperimmune serum was raised against it, which was used later while developing dot-ELISA. The dot-ELISA exhibited higher specificity (92%) in diagnosing MD lymphomas as compared to conventional PCR (40%), where later assay is unable to differentiate disease development (lymphoma) and/or infection. The developed dot-ELISA proved to be a specific, rapid and inexpensive technique detecting MDV lymphomas in poultry. Of the note, this new assay could be opted as a valuable diagnostic tool in the resource poor countries andcould further be used to differentiate from other tumor causing viruses in poultry.

Effects of aortic root motion on wall stress in the Marfan aorta before and after personalised aortic root support (PEARS) surgery.

Aortic root motion was previously identified as a risk factor for aortic dissection due to increased longitudinal stresses in the ascending aorta. The aim of this study was to investigate the effects of aortic root motion on wall stress and strain in the ascending aorta and evaluate changes before and after implantation of personalised external aortic root support (PEARS). Finite element (FE) models of the aortic root and thoracic aorta were developed using patient-specific geometries reconstructed from pre- and post-PEARS cardiovascular magnetic resonance (CMR) images in three Marfan patients. The wall and PEARS materials were assumed to be isotropic, incompressible and linearly elastic. A static load on the inner wall corresponding to the patients' pulse pressure was applied. Cardiovascular MR cine images were used to quantify aortic root motion, which was imposed at the aortic root boundary of the FE model, with zero-displacement constraints at the distal ends of the aortic branches and descending aorta. Measurements of the systolic downward motion of the aortic root revealed a significant reduction in the axial displacement in all three patients post-PEARS compared with its pre-PEARS counterparts. Higher longitudinal stresses were observed in the ascending aorta when compared with models without the root motion. Implantation of PEARS reduced the longitudinal stresses in the ascending aorta by up to 52%. In contrast, the circumferential stresses at the interface between the supported and unsupported aorta were increase by up to 82%. However, all peak stresses were less than half the known yield stress for the dilated thoracic aorta.

Biomechanical properties of the Marfan's aortic root and ascending aorta before and after personalised external aortic root support surgery.

Marfan syndrome is an inherited systemic connective tissue disease which may lead to aortic root disease causing dilatation, dissection and rupture of the aorta. The standard treatment is a major operation involving either an artificial valve and aorta or a complex valve repair. More recently, a personalised external aortic root support (PEARS) has been used to strengthen the aorta at an earlier stage of the disease avoiding risk of both rupture and major surgery. The aim of this study was to compare the stress and strain fields of the Marfan aortic root and ascending aorta before and after insertion of PEARS in order to understand its biomechanical implications. Finite element (FE) models were developed using patient-specific aortic geometries reconstructed from pre and post-PEARS magnetic resonance images in three Marfan patients. For the post-PEARS model, two scenarios were investigated-a bilayer model where PEARS and the aortic wall were treated as separate layers, and a single-layer model where PEARS was incorporated into the aortic wall. The wall and PEARS materials were assumed to be isotropic, incompressible and linearly elastic. A static load on the inner wall corresponding to the patients' pulse pressure was applied. Results from our FE models with patient-specific geometries show that peak aortic stresses and displacements before PEARS were located at the sinuses of Valsalva but following PEARS surgery, these peak values were shifted to the aortic arch, particularly at the interface between the supported and unsupported aorta. Further studies are required to assess the statistical significance of these findings and how PEARS compares with the standard treatment.

Clinical profile and antibiotics sensitivity in childhood urinary tract infection at Dhulikhel Hospital.

BACKGROUND: Urinary Tract Infection implies presence of actively multiplying organisms in the urinary tract. Although it is infrequently associated with mortality, it is still a significant cause of morbidity. Early diagnosis is critical to preserve renal function of growing kidney. OBJECTIVE: Our purpose was to determine the clinical, microbiologic profile and antibiotic sensitivity of such infections in pediatric Urinary Tract Infection (UTI) patients at Dhulikhel Hospital. METHODS: A hospital based prospective descriptive study of 135 children from 2 months to 16 years, with clinical diagnosis of urinary tract infection who visited the pediatric department of Dhulikhel Hospital over the period of 15 months were enrolled in the study. All patients underwent routine urine analysis and culture. Children with recurrent UTI underwent micturating cystourethrogram (MCUG). Children with recurrent UTI of more than two years and with feature of pyelonephritis underwent USG abdomen as well. Complications and response of the treatment was observed in all cases of UTI. All data were entered in Epidata and data analysis was done using spss 16 version. RESULTS: Among 135 children, 32.5% were male and 67.4% were female. Fever was the most common presenting symptom in 74.80% of patients followed by dysuria in 54.1%. Among these children 95.6% had significant pyuria and 45% had culture positive infection. Children who showed positive for bacteriuria, Escherichia coli (78.7%) was the most common organism and are more than 80% sensitive to Amikacin, Gentamicin, Ceftriaxone, Ofloxacin, Nalidixic acid, Imipenem and Vancomycin. Co-trimoxazole was the most common drug used for treatment with a mean drug respond time of (mean+/-S.D) of 2.21+/-.78 days. 2+/-. Children who had recurrent UTI were more prone to develop culture positive UTI (p=0.0001). CONCLUSION: Urinary Tract Infection in female was almost twice more common than in male. Cotrimoxazole was the most common drug used for treatment, sensitivity of this drug was less than 50% for all organisms.

Fowl adenovirus (FAdV) in India: evidence for emerging role as primary respiratory pathogen in chickens.

Adenoviruses have been isolated from both clinically healthy and diseased birds worldwide. The pathogenic role of most of the FAdVs is still questionable. They can quickly take on the role of opportunistic pathogens when additional factors, particularly concurrent infections, adversely affect the health of the avian host. Immnosuppressing agents especially chicken infectious anemia and infectious bursal disease viruses are known to enhance the pathogenicity of FAdVs upon coinfection. The aim of the present study was to screen for the involvement of FAdV in poultry flocks affected with respiratory disease complex by RT-PCR. The samples were also screened by RT-PCR/PCR for other respiratory pathogens. Thirty two commercial poultry flocks with the history of respiratory disease complex from various parts of India. FAdV nucleic acid could be detected in tissue samples of 13 out of 34 farms investigated. Out of 13 FAdV positive farms, FAdV and CIAV were alone detected in 4/13 (31%) whereas, in other farms more than two respiratory pathogens were detected together. CIAV was detected in all the farms (34/34) investigated. Eosinophilic intranuclear inclusion bodies were noticed in FAdV infected laryngeal and tracheal epithelium under light microscopy. The findings of the study assert that FAdV can play the role of primary respiratory pathogen in immunocompromised birds and also in the presence of other respiratory pathogens.

Increased T-regulatory cells within lymphocyte follicles in moderate COPD.

Lymphoid follicles in the lung parenchyma are a characteristic feature of chronic obstructive pulmonary disease (COPD). There are reports of altered CD4 T-regulatory cell numbers in COPD lungs, but the location of these cells within COPD lung tissue specific follicles has not been investigated. The presence of CD4(+)FOXP3(+) T-regulatory cells was assessed in surgically resected lung tissue from 12 COPD patients, 11 smokers with normal lung function and seven nonsmokers by combined immunofluorescence and immunohistochemistry. Organised lymphoid follicles were observed in all three groups of patients, as well as lymphoid clusters lacking organisation. The percentage of CD4 cells that were T-regulatory cells were significantly increased (p = 0.02) within COPD (16%) follicles compared with smokers (10%) and nonsmokers (8%). In contrast, there was no change (p>0.05) in the percentage of T-regulatory cells in clusters or the subepithelium between groups. Lymphoid follicles in COPD patients have increased T-regulatory cells. Therefore, T-regulatory activity may be altered within COPD lymphoid follicles.

Comparative evaluation of wound healing potency of Vitex trifolia L. and Vitex altissima L.

The wound healing potency of ethanol leaf extracts of V. trifolia L. and V. altissima L. was evaluated in excision, incision and dead space wound models. Both plants were found to possess significant wound healing activity which was evidenced by a decrease in the period of epithelialization, an increase in the rate of wound contraction, skin breaking strength, granulation tissue dry weight, hydroxyproline content and breaking strength of granulation tissue. Histopathological study of the granulation tissue also showed an increased collagenation when compared with the control group of animals. Of the two extracts, the ethanol leaf extract of V. trifolia showed maximum wound healing activity compared with the leaf extract of V. altissima. However, on comparison with the control group, both leaf extracts were found to possess significant wound healing potency.

Immunosuppression in broiler chicks fed aflatoxin and inoculated with fowl adenovirus serotype-4 (FAV-4) associated with hydropericardium syndrome.

A total of 240 unvaccinated day-old broiler chicks, which had been found to be negative for antibodies against FAV-4, were divided into four groups of 60 chicks each. Group A was fed aflatoxin at 1 ppm from 7 days to 7 weeks of age. Group V was infected intra-abdominally at 14 days of age with 0.2 ml of FAV-4, having a titre of 10(5.5) TCID50 per 0.2 ml. The combined group AV was given the aflatoxin and infected with FAV-4. The fourth group C served as the control. More pronounced clinical signs, a higher mortality rate (56.7%), and reductions in body weight gain and in the organ to body weight ratios of the bursa and spleen were recorded in group AV. A significant (p < 0.01) reduction in the HI antibody titre following vaccination against Newcastle disease, and of skin thickness in the delayed hypersensitivity test following sensitization with DNCB, indicated an additive immunosuppressive effect from aflatoxin and FAV-4 on the humoral and cell-mediated immune responses in group AV compared to groups A and V. Microscopically, marked depletion and degeneration of lymphocytes in the thymus, bursa, spleen and caecal tonsils were observed in group AV up to 5 weeks PI.

Familial cardiac myxoma--a comprehensive review of reported cases.

BACKGROUND: Cardiac myxoma with familial involvement is a rare manifestation of an uncommon tumor, which nevertheless has been reported with increasing frequency in the past two decades. As a feature of the hereditary Carney syndrome, cardiac myxoma occurs at a much younger age, often has multicentric origins, and tends to recur. We report the case of a family in which three members had cardiac myxoma and two had other characteristics of the Carney syndrome. METHODS: We have made a comprehensive review of the international literature from 1971-1992 to determine the incidence and clinical presentation of cardiac myxoma when diagnosed in more than one family member or first-degree relative. RESULTS: Twenty-six cases of familial cardiac myxoma, involving 68 family members, have now been reported in the United States, Europe, and Australia. One-fourth of these reports have appeared in the past 4 years alone. The rate of diagnosis at autopsy has declined from 40% in early reports to a current 17%. Seventeen episodes of recurrence (25%) at the same or different intracardiac sites have been documented, approximately twice the rate of recurrence in isolated cases. CONCLUSIONS: The diagnostician should be alert for evidence of cutaneous, endocrine, or testicular features of the hereditary Carney syndrome in young patients who have cardiac myxoma. In the families of these patients, all first-degree relatives should be examined regularly for evidence of cardiac myxoma.

The use of ventilation tubes and the incidence of cholesteatoma surgery in the paediatric population of Liverpool.

Many surgeons believe that grommet insertion has reduced the incidence of cholesteatoma and thus radical mastoidectomy. Several causes for this trend have been postulated, including the increased use of ventilation tubes to treat otitis media with effusion. We have studied the number of operations for cholesteatoma, together with the number of ventilation tube insertions in the paediatric population of Liverpool, over a 28-year period between 1963 and 1990. There was a decline in the number of operations for cholesteatoma and an increase in the use of ventilation tubes, but there was no significant correlation between the two. This suggests that other factors are responsible for the decline in surgery for cholesteatoma.

Clinicopathological studies of carcinoma oesophagus with special reference to changes in surrounding epithelium.

A total 17 cases of carcinoma oesophagus were studied in a period of 2 years. Ten patients (58.82%) were males and rest were females. Maximum cases (64.70%) were seen in 5th and 6th decades. Majority of the patients belonged to low socio-economic group (47.05%) followed by middle socio-economic group (41.17%). Dysphagia was present in all cases, followed by weight loss 52.94% and other symptoms. Cancer was equally distributed in middle third and lower third of the oesophagus (41.18% in each group) only in 3 cases it was in upper third of the oesophagus. Histologically 70.5% tumours were squamous cell carcinoma, 17.64% were adenocarcinoma and 11.76% were undifferentiated carcinoma. Surrounding epithelium in 17.64 per cent cases showed chronic oesophagitis, 4 cases (23.52%) showed acanthosis and dysplasia, and two cases revealed carcinoma in situ. Hence findings of oesophagitis, acanthosis, dysplasia, carcinoma in situ suggest that oesophagitis and acanthosis may be considered as precancerous lesions.

Pre-cancerous lesions of stomach.

Total 39 cases of carcinoma stomach were noticed out of 142 malignant tumours of GIT (27.46 percent). Histologically maximum cases were of diffuse type (56.41 percent) followed by intestinal type (35.89 percent) and indolent mucoid carcinoma (7.69 percent) of the stomach. The surrounding epithelium showed lot of changes in the intestinal type of carcinoma stomach. About 78.57 percent showed intestinal metaplasia, 14.28 percent of these cases showed chronic gastric ulcer and severe dysplasia (carcinoma in situ) and another 14.28 percent revealed villous adenoma with carcinoma in situ. In contrast to this, in diffuse variety, only 13.63 percent cases revealed intestinal metaplasia, 27.27 percent showed basal cell hyperplasia, stratification of the epithelium of crypts and diffuse infiltration of mucosa by malignant cells and 4.54 percent showed atrophic gastritis also. In mucoid carcinoma all cases had basal cell hyperplasia and stratification of crypts. Hence these conditions should be taken as premalignant lesions of stomach and should be cured in proper time.

The presentation and management of the nasal glioma.

Nasal gliomata are rare and experience in management is therefore not widespread. Clinical details of 5 children with nasal gliomata are presented. In each case the glioma was excised by an external approach without the need for craniotomy. These cases demonstrate the variability of presenting features and problems in management. They illustrate a relatively conservative surgical approach to treatment which we consider to be safe and effective. The characteristics of these unusual neural malformations are reviewed, and the management is discussed.

The fixed cervical lymph node.

A series of almost 2000 patients with squamous carcinoma of the head and neck is presented. Seven percent of the patients had massive fixed glands at the time of presentation. The incidence of fixed nodes varied between sites, and increased with increasing T-stage of the primary tumor. Forty percent of the patients were treated; the most common cause for withholding treatment was advanced age. Radiotherapy did not produce a significant prolongation of survival, but surgery did. Resection of the mandible, the skin of the neck, and the external carotid artery proved to be valuable procedures both in terms of palliation and survival; radical surgery produced a 5-year survival of approximately 15%.

Mortality after surgery for hypopharyngeal cancer.

Results of 362 patients with squamous carcinoma of the hypopharynx are presented. The hospital mortality was 25 per cent for patients treated surgically. The mortality was increased by previous radiotherapy and poor general condition, and also depended on the method of pharyngeal repair: the mortality for visceral (stomach or colon) transposition was higher than that for repair by skin flaps. The chance of long term survival decreased with increasing N stage, and was also reduced for patients undergoing visceral repair compared with skin flap repair of the pharynx.