Study of knowledge, attitude, and practice regarding hepatitis B infection in first-degree relatives of patients suffering with hepatitis B infection

Background: Hepatitis B virus (HBV) infection is a significant public health problem in third-world countries, including India. The risk of infection among close family members of patients infected with chronic HBV infection is four to six times higher than in the general population. However, literature is scarce regarding the Knowledge, attitude, and practice toward HBV infection among the first-degree relatives of patients suffering from chronic hepatitis B. Hence, this study aimed to assess the knowledge, attitude, and practice regarding hepatitis B virus infection among first-degree relatives of hepatitis B patients. Methods: A cross-sectional, observational study was conducted amongst the 354 first-degree relatives of HBsAg-positive patients. A validated questionnaire consisted of twenty questions of knowledge, seven questions of attitude, and ten practice questions were employed to gather information. Results: The mean knowledge, attitude, and practice (KAP) were significantly better amongst participants aged 55-65 years and those who were educated. There was a positive correlation between knowledge and attitude (r=0.58, p value<0.05) and knowledge and practice (r=0.64, p value<0.05). Conclusions: There is insufficient knowledge regarding hepatitis B, its common symptoms, modes of transmission, and preventive tools among close relatives of patients suffering from chronic hepatitis B.

Effect of Cage or Deep Litter Housing on Production Performance of White Leghorn Chickens

The effect of cage or deep litter housing on production performance of White Leghorn was investigated in this experiment. A total 144 pullets of White Leghorn were randomly assigned to two treatments as; Cage housing and Deep litter housing. Results revealed that body weight at sexual maturity was significantly (P<0.05) lower in birds reared in cage than the deep litter housing. Moreover, lower feed intake was observed in birds kept in cages than that of deep litter housing. Feed conversion ratio in terms of feed intake/dozen eggs or feed intake/kg egg mass was better in birds reared in cages than the deep litter housing. The average hen day and hen house egg production were not affected due to different housing systems. Egg quality traits like albumen index, yolk index and Haugh unit score were significantly (P<0.05) higher for eggs produced by birds reared in cage housing than the deep litter housing. The profit of about 20% was observed in cage housing over deep litter housing. It was concluded that the lower feed consumption and body weight at sexual maturity, better feed conversion, egg quality and economic returns were achieved in cages compared to deep litter housing. However, cage or deep litter housing had no effect on egg production

Development of immunodetection system for botulinum neurotoxin serotype E

Background & objectives: Botulism, a potentially fatal paralytic illness, is caused by the botulinum neurotoxins (BoNTs) secreted by Clostridium botulinum. It is an obligate anaerobic, Gram-positive, spore-forming bacterium. BoNTs are classified into seven serotypes based on the serological properties. Among these seven serotypes, A, B, E and, rarely, F are responsible for human botulism. The present study was undertaken to develop an enzyme-linked immunosorbent assay (ELISA)-based detection system for the detection of BoNT/E. Methods: The synthetic gene coding the light chain of BoNT serotype E (BoNT/E LC) was constructed using the polymerase chain reaction primer overlapping method, cloned into pQE30UA vector and then transformed into Escherichia coli M15 host cells. Recombinant protein expression was optimized using different concentrations of isopropyl-?-D-1-thiogalactopyranoside (IPTG), different temperature and the rBoNT/E LC protein was purified in native conditions using affinity column chromatography. The purified recombinant protein was checked by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and further confirmed by western blot and matrix-assisted laser desorption ionization-tandem time-of-flight (MALDI-TOF). Polyclonal antibodies were generated against rBoNT/E LC using Freund's adjuvant in BALB/c mice and rabbit. Sandwich ELISA was optimized for the detection of rBoNT/E LC and native crude BoNT/E, and food matrix interference was tested. The developed antibodies were further evaluated for their specificity/cross-reactivity with BoNT serotypes and other bacterial toxins. Results: BoNT/E LC was successfully cloned, and the maximum expression was achieved in 16 h of post-induction using 0.5 mM IPTG concentration at 25癈. Polyclonal antibodies were generated in BALB/c mice and rabbit and the antibody titre was raised up to 128,000 after the 2nd booster dose. The developed polyclonal antibodies were highly specific and sensitive with a detection limit about 50 ng/ml for rBoNT/E LC and 2.5�[3] MLD50 of native crude BoNT/E at a dilution of 1:3000 of mouse (capturing) and rabbit (revealing) antibodies. Further, different liquid, semisolid and solid food matrices were tested, and rBoNT/E LC was detected in almost all food samples, but different levels of interference were detected in different food matrices. Interpretation & conclusions: There is no immune detection system available commercially in India to detect botulism. The developed system might be useful for the detection of botulinum toxin in food and clinical samples. Further work is in progress.

Structural and functional analysis of chitinase gene family in wheat (Triticum aestivum).

Chitinases are the hydrolytic enzymes which protect plants against pathogen attack. However, the precise role of chitinases in disease resistance has not been explored in wheat. In the present study, in silico approach, including secondary structure analysis, detailed signature pattern study, cis-acting regulatory elements survey, evolutionary trends and three-dimensional molecular modeling was used for different chitinase classes of wheat (Triticum aestivum). Homology modeling of class I, II, IV and 3 chitinase proteins was performed using the template crystal structure. The model structures were further refined by molecular mechanics methods using different tools, such as Procheck, ProSA and Verify3D. Secondary structure studies revealed greater percentage of residues forming α helix conformation with specific signature pattern, similar to casein kinase II phosphorylation site, amidation site, N-myristoylation (N-MYR) site and protein kinase C phoshorylation site. The expression profile suggested that wheat chitinase gene was highly expressed in cell culture and callus. We found that wheat chitinases showed more functional similarity with rice and barley. The results provide insight into the evolution of the chitinase family, constituting a diverse array of pathogenesis-related proteins. The study also provides insight into the possible binding sites of chitinase proteins and may further enhance our knowledge of fungal resistance mechanism in plants.

Clinical and microbiological characteristics of Rhodotorula mucilaginosa infections in a tertiary-Care facility.

Background: Rhodotorula spp. are an emergent opportunistic pathogen, particularly in immunocompromised individuals. Materials and Methods: The aim of the study was to review reported cases of Rhodotorula infection over a period of 9 years to determine epidemiology, risk factors, treatment and outcome. Results: The Rhodotorula spp. were isolated from cerebrospinal fluid (9) and blood (5). The most common pre-disposing factors were prolonged hospital stay (>1 month) and prolonged usage of broad-spectrum antibiotics (>1 month). All the isolates were identified as R. mucilaginosa by conventional methods. Amphotericin B demonstrated lowest minimum inhibitory concentration (MIC) as compared with other anti-fungal agents (fluconazole, itraconazole and voriconazole). Conclusions: The recognition of unusual yeasts as an agent of life-threatening infection and their intrinsic resistance increases the burden on the mycology laboratory for complete species identification and to determine minimum inhibitory concentration.

Foreign bodies in upper aero-digestive tract: a clinical study.

Background: Foreign-body ingestion and aspiration are common childhood adverse events, hence are commonest causes of morbidity and mortality in infants and children worldwide. Aim of current study was to identify the patients with foreign-body ingestion and aspiration, develop a suitable algorithm for their management and study various complications following their removal. Methods: The present study was carried out in G.R. medical college, Gwalior, M. P. during last one year on hundred patients diagnosed as case of foreign-body in upper aero-digestive tract on the basis of history, examination and investigations. Results: Foreign-bodies in upper aero-digestive tract were seen mostly in children less than 10 years of age (91%). The youngest was 8 months old and the oldest 48 years. Coins were the most common foreign-bodies in oesophagus (90%) while whistle in the upper airway (40%). Right bronchus is the commonest site of impaction in the airway (50%), while cricopharynx is most common in the oesophagus (95.55%). Patients with oesophageal foreign bodies presented mostly with the chief complaint of foreign body sensation (63.4%) in the throat whereas those with airway foreign body presented mostly with dyspnoea and choking sensation (60%). Pooling of saliva was most commonly seen in patients with oesophageal foreign bodies (43.3%). Tachypnoea was the most common sign in cases of tracheabronchial foreign bodies (80%). Positive radiological evidence was present in 88% of upper aero-digestive foreignbodies. Only 2.2% cases showed complications following oesophagoscopy. Tracheo-bronchial foreign-bodies were removed by emergency bronchoscopy. 20% cases had complications post-operatively. Conclusion: Early detection by meticulous history, imaging modality & prompt management remains basis for favourable outcome and prevents fatal complications.

A prospective study for comparison of hematological parameters in healthy young adult and elderly age group subjects.

Aims & Objectives: This study of hematological parameters is the object of continuing intensive research of hematopoietic system to the functional integrity in aging. (1) Assessment of Hb, total RBC count in healthy young adult as well as in elderly subject.(2) Assessment of Hct & blood indices (MCV, MCH, MCHC) in healthy young adult as well as in elderly subject.(3) Comparing the values of Hb & total RBC count in healthy young adult with the elderly subject.(4) Comparing the values of Hct & blood indices (MCV, MCH, MCHC) in healthy young adult with the elderly subject.(5) To co-relate the results of present study with findings of other studies. Material & Methods: The hematological indices analysed were Hb, HCT, and Total RBC Count, Blood indices like MCV, MCH & MCHC. Hematological values were measured by the different method in laboratory with the use of standard test reagent. Results: In present study, the result shows that a significant decrease in Hb concentration and RBC count are associated with decrease in Hct, MCH, MCHC and little increase in MCV. Conclusion: In present study, the overall assessment shows that a significant decrease in Hb concentration, reticulocyte count and RBC count are associated with decrease in Hct, MCH, MCHC and little increase in MCV. These results show a significant decrease in haematological parameters in old age and fairly accurate reflection of decreased erythropoietic activity of bone marrow.

A comparative study of reticulocyte count in healthy young adult and elderly age group subjects.

Introduction: Aging process has been viewed as progressive decline in physiological process necessary for maintenance of a constant milieu interior and death as the ultimate failure to sustain homeostasis. Reticulocytes are juvenile red cell. It is flat non-nucleated disc shaped, slightly larger than mature erythrocyte .The number of reticulocyte present in the peripheral blood is a fairly accurate reflection of erythropoitic activity of bonemarrow. Normal values in young adult- 0.5 to 2.5% of total RBC count. The present study was conducted to find out changes in the reticulocyte count in the elderly subject. Aims: to uncover the effect of aging on reticulocyte count. Material and method: 90 subjects were divided into two groups, normal control group (n=30, mean age 35.83yrs) and elderly group (n=60, mean age62.55yrs). Reticulocyte counts were done by new Methylene Blue method. Result & Observations: significant decrease in reticulocyte count was observed in elderly group as compare to normal control group. Conclusion: our data suggests that aging may alter haematological parameters including reticulocyte count.

Periampullary carcinoid: An uncommon tumor at an unusual site.

Gastrointestinal carcinoid tumors represent a group of well-differentiated tumors originating from various neuroendocrine cells located in the gastrointestinal mucosa and submucosa. Consequently, there is diversity in their clinical presentation, incidence at specific anatomic sites, biological behavior, hormone production, morphologic characteristics, and immunophenotype. Periampullary carcinoids are extremely rare and less then 100 patients have been reported in the world literature, that too mostly as case reports. We are reporting two cases of periampullary carcinoids, one of which presented with rare manifestation as gastrointestinal bleed and both are doing well after successful pancreatoduodenectomy.

Avian human influenza : diagnosis, case management and treatment.

The H5N1 influenza virus infected human in 1997 for the first time in Hong Kong leading to development of influenza like symptoms. Its incubation period is 8-17 days. Cases of influenza can be confirmed with possible viral culture for H5N1 or positive PCR for H5N1 or positive IFA test using monoclonal antibodies against H5N1 or a four-fold rise in H5N1 specific antibody titre. The plain x-ray chest may show patchy opacities involving at least two zones, lobar consolidation with air and ultimately rapidly evolving ground glass appearance may occur. WHO recommends use of oseltamivir for case management. Public health measures include to report all suspected cases of influenza and to identify all cases might have been exposed to common source of infection and they should be monitored for 7 days.

Management of chronic hepatitis B with New Livfit in end stage renal disease.

New Livfit (NLF) is a standardized, poly-herbal formulation that has been found useful in the management of hepatitis. The aim of this placebo-controlled study was to evaluate its usefulness against hepatitis B virus in the patients of end stage renal disease (ESRD). Patients were regularly evaluated at 6, 12, 24 and 36 weeks of therapy. With 36 weeks of treatment of NLF, there was rapid clearing of HBV-DNA in a significant number of patients. Significant seroconversion of the other markers of hepatitis B and restoration of the raised levels of ALT and AST was observed. The study suggests the potential usefulness of NLF in the control of HBV infection in the patients of ESRD prior to renal transplant.

Ultra-structural observations on the lymphoid organs of the freshwater catfish, Clarias batrachus (Linnaeus).

Light microscopic and ultra-structural studies of the lymphoid tissues such as blood immunocytes, spleen and pronephros of the freshwater catfish, Clarias batrachus, were carried out. The peripheral blood showed nucleated erythrocytes, total leucocytic count (TLC) more than that observed in mammalian blood and leucocytes with morphological appearance similar to the mammalian white blood cells (WBCs). The spleen and pronephros showed presence of numerous lymphocytes, monocytes and nucleated red blood cells (RBCs) along with hemosiderin-containing macrophages. The morphology of lymphoid organs of the catfish has been discussed in light of the evolution of the immune system in this class of vertebrates.

Hepatorenal syndrome.

Hepatorenal syndrome is a life-threatening complication occurring commonly in cirrhosis liver and rarely in acute liver failure. It can be precipitated by shock, infection, surgery, large volume paracentesis or nephrotoxic drugs. Type I hepatorenal syndrome which usually develops over acute liver failure is rapidly progressive and has poor outcome. Type II hepatorenal syndrome is usually associated with refractory ascites and is slowly progressive with relatively good prognosis. Peripheral vasodilation with intrarenal vasoconstriction is the main pathophysiologic change. Diagnostic criteria, ascertained by international ascites club, is helpful to reach at a proper diagnosis. Management includes pharmacologic therapy to induce splanchnic vasoconstriction which improve renal circulation. Dopamine, vasopressin analogs (ornipressin and terlipressin), midodrine, noradrenaline have been used mainly as a bridge to the liver transplant or in anticipation of improvement in hepatic function. The molecular adsorbent recycling system (MARS) have been recently used in patients with hepatorenal syndrome. Transjugular intrahepatic portosystemic shunt is also another modality which has been used as a bridge to liver transplant in such patients.

14C-urea breath test for assessment of gastric Helicobacter pylori colonization and eradication.

BACKGROUND AND OBJECTIVE: Urea breath test (UBT) is a reliable noninvasive technique for detecting gastric Helicobacter pylori colonization. 14C isotope-based test requires simple equipment and is inexpensive. We studied the utility of 14C-UBT in diagnosis of gastric H. pylori infection. METHODS: Presence of H. pylori was studied using antral histology and culture in patients with rapid urease test (RUT)-positive peptic ulcer. 14C-UBT was performed using a 185-kBq dose. Radioactivity in 15-min breath samples was measured using a beta-scintillation counter and result expressed as % dose recovered/mmol CO2. H. pylori was considered positive when any two tests were positive. All tests were repeated one month after completion of H. pylori eradication therapy. RESULTS: Among 41 patients (duodenal ulcer 36, gastric ulcer 5), H. pylori was detected by histology in 23 (56%) and by culture in 27 (66%). Overall, H. pylori was detected in 28 (68%) patients. Follow-up assessment was possible in 28 patients: 26 cleared the infection (all three tests negative). Mean 14C recovery values at 15 minutes associated with H. pylori-positive status were significantly higher (12.3 [SD 6.8] x 10(-3); n=30; p<0.001) than those associated with H. pylori-negative status (2.1 [0.9] x 10(-3); n=26). Using receiver-operating-characteristic analysis of 15-minute 14C recovery values, a cut-off of 6.5x10(-3) gave the best separation of H. pylori-positive and -negative cases. 14C-UBT had 93% sensitivity, 96% specificity and 95% accuracy. CONCLUSION: 14C-UBT appears to be a reliable noninvasive test for diagnosis of H. pylori infection.

Study of events leading cellular senescence to human mammary epithelial cancer cells in vitro.

Role of various growth regulatory factors in inducing senescence in cultured HMEC cells have been investigated in ten cases of breast cancer. The histological grade of tumour cells is found to play significant role in controlling the proliferation and phenotypic charateristics of cultured HMEC cells during primary culture and also number of subsequent passages resulting in complete cellular senescence in them. Effects of conditioned media (CM) collected from primary and senescent cultures of these HMEC cells had also been studied on proliferation of their own HMEC cells used as target cells, to evaluate the role of various autocrine growth factors produced by them. Significant increase in proliferation of target cells was noticed on their exposure to CM from senescent cultures, while cessation of their proliferation was found on their exposure to CM from senscent cultures, suggesting that HMEC cells produce growth promoting factors during primary culture and growth inhibitory factors on subsequent passages, responsible for inducing features of cellular senescence in them. The role of epidermal growth factors (EGF) and transforming growth factors (TGF) alpha and beta as autocrine factors in inducing senescence of cultured HMEC cells were also investigated. Deletion of EGF from growth media initially caused decreased proliferation to target HMEC cells, followed by improvement in their proliferation. Supplementation of growth media by TGF-alpha induced significant increase in proliferation of target cells. Addition of epidermal growth factors receptor (EGFR) antibody to cells exposed to media devoid of EGF and media supplemented with TGF-alpha showed marked suppression of proliferation of target cells. The morphologic and phenotypic characteristics of target HMEC cells exposed to TGF-alpha were also found similar to those HMEC cells grown during primary culture, suggesting autocrine production of EGF and TGF-alpha by cultured HMEC cells during primary culture. Supplementation of TGF-beta to growth media induced marked suppression of proliferation to target cells along with morphologic and phenotypic features of terminal differntiation or senescence. Exposure of senescent cells to media supplemented with EGF and TGF-alpha could not induce their proliferation. This suggest that HMEC cells on subsequent passages undergo some genetic and phenotypic alterations resulting in production of growth inhibitory factor like TGF-beta which induces cessation of their proliferation alone with features of senescence.