Isolation and genome analysis of a lytic Pasteurella multocida Bacteriophage PMP-GAD-IND.

Currently used alum precipitated and oil adjuvant vaccines against HS caused by Pasteurella multocida B:2, have side effects and short-lived immunity, leading to regular catastrophic outbreaks in bovines in Asian subcontinent. The need for the development of an improved vaccine with longer immunity and the ability to differentiate between vaccinated and infected is essential. Pasteurella phage isolated in present study belongs to family Siphoviridae. PMP-GAD-IND phage exhibited lytic activity against vaccine strain (P52) as well as several field strains of P. multocida (B:2), and fowl cholera agent (P. multocida A:1).The phage has a double stranded DNA (dsDNA) with a genome of 46 335 bp. The complete genome sequence of the Pasteurella multocida phage has been deposited in Gen Bank with accession no: KY203335. PMP-GAD-IND being a lytic phage with broad activity range has a potential to be used in therapy against multidrug resistant P. multocida infections. SIGNIFICANCE AND IMPACT OF THE STUDY: The present work is a part of research for the development of an improved phage lysate marker vaccine and a companion DIVA assay against haemorhagic septicaemia. This study describes the isolation and genome analysis of PMP-GAD-IND a lytic Pasteurella multocida bacteriophage.

X-ray Reflectivity Study of the Interaction of an Imidazolium-Based Ionic Liquid with a Soft Supported Lipid Membrane.

Ionic liquids (ILs) are important for their antimicrobial activity and are found to be toxic to some microorganisms. To shed light on the mechanism of their activities, the interaction of an imidazolium-based IL 1-butyl-3-methylimidazolium tetrfluoroborate ([BMIM][BF4]) with E. coli bacteria and cell-membrane-mimicking lipid mono- and bilayers has been studied. The survival of the bacteria and corresponding growth inhibition are observed to be functions of the concentration of the IL. The IL alters the pressure-area isotherm of the monolayer formed at an air-water interface by the 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) lipid. The in-plane elasticity of the lipid layer is reduced as a consequence of the insertion of this IL. The X-ray reflectivity study from a polymer-supported lipid bilayer shows strong perturbation in the self-assembled structure of the bilayer due to the interaction. As a consequence, there is a considerable decrease in bilayer thickness and a corresponding increase in electron density. These results, however, depend on the chain configurations of the lipid molecules.

Protective immune-response of aluminium hydroxide gel adjuvanted phage lysate of Brucella abortus S19 in mice against direct virulent challenge with B. abortus 544.

The prophylactic efficacies of plain and alum adsorbed lysate were evaluated by direct virulent challenge in mice model. A recently isolated brucellaphage 'ϕLd' was used for generation of lysates. Twenty four h incubated Brucella abortus S19 broth cultures standardized to contain approximately 10(8) CFU/ml were found suitable for generation of lysates. Three lysate batches produced through separate cycles did not show any significant variation with respect to protein and polysaccharide contents, endotoxin level and phage counts, indicating that compositionally stable lysate preparations can be generated through an optimized production process. Three polypeptides of ∼16, 19 and 23 kDa could be identified as immuno-dominant antigens of the lysate which induced both humoral and cell-mediated immune responses in a dose dependent manner. Results of efficacy evaluation trial confirmed dose-dependent protective potencies of lysate preparation. The lysate with an antigenic dose of 0.52 µg protein and 60 µg CHO adsorbed on aluminium gel (0.1 percent aluminium concentration) exhibited the highest protective potency which was greater than that induced by standard S19 vaccine. Phage lysate methodology provides a very viable option through which an improved immunizing preparation with all desirable traits can be developed against brucellosis, and integrated with immunization programmes in a more efficient manner.

The GPCR OGR1 (GPR68) mediates diverse signalling and contraction of airway smooth muscle in response to small reductions in extracellular pH.

BACKGROUND AND PURPOSE: Previous studies have linked a reduction in pH in airway, caused by either environmental factors, microaspiration of gastric acid or inflammation, with airway smooth muscle (ASM) contraction and increased airway resistance. Neural mechanisms have been shown to mediate airway contraction in response to reductions in airway pH to < 6.5; whether reduced extracellular pH (pHo) has direct effects on ASM is unknown. EXPERIMENTAL APPROACH: Intracellular signalling events stimulated by reduced pHo in human cultured ASM cells were examined by immunoblotting, phosphoinositide hydrolysis and calcium mobilization assays. ASM cell contractile state was examined using magnetic twisting cytometry. The expression of putative proton-sensing GPCRs in ASM was assessed by real-time PCR. The role of ovarian cancer G protein-coupled receptor 1 (OGR1 or GPR68) in acid-induced ASM signalling and contraction was assessed in cultures subjected to siRNA-mediated OGR1 knockdown. KEY RESULTS: ASM cells responded to incremental reductions in pHo (from pH 8.0 to pH 6.8) by activating multiple signalling pathways, involving p42/p44, PKB, PKA and calcium mobilization. Coincidently, ASM cells contracted in response to decreased pHo with similar 'dose'-dependence. Real-time PCR suggested OGR1 was the only proton-sensing GPCR expressed in ASM cells. Both acid-induced signalling (with the exception of PKB activation) and contraction were significantly attenuated by knockdown of OGR1. CONCLUSIONS AND IMPLICATIONS: These studies reveal OGR1 to be a physiologically relevant GPCR in ASM cells, capable of pleiotropic signalling and mediating contraction in response to small reductions in extracellular pH. Accordingly, ASM OGR1 may contribute to asthma pathology and represent a therapeutic target in obstructive lung diseases.

Complete migration of ventriculoperitoneal shunt into the ventricle.

Proximal migration of ventriculoperitoneal (VP) shunt is extremely rare. A medium pressure VP shunt was inserted in a 10-month-old infant suffering from severe hydrocephalus. One month later, complete migration of the shunt into the lateral ventricle was encountered. Possible causative mechanisms and the literature on this subject are briefly discussed.

Comparison of continuous thoracic epidural and paravertebral blocks for postoperative analgesia after minimally invasive direct coronary artery bypass surgery.

OBJECTIVE: To compare continuous thoracic epidural analgesia (TEA) and paravertebral block (PVB) for postoperative analgesia in patients undergoing minimally invasive direct coronary artery bypass (MIDCAB) surgery for quality of analgesia, complications, compliance to chest physiotherapy, hemodynamics, and respiratory effects. DESIGN: Prospective, randomized study. SETTING: Specialty research hospital. PARTICIPANTS: Forty-one consenting patients undergoing MIDCAB surgery. INTERVENTIONS: Patients in the TEA group had an epidural catheter inserted at the T4-5 interspace, whereas patients in the PVB group had a catheter inserted in the paravertebral space on the left side at the T4-5 level. MEASUREMENTS AND MAIN RESULTS: Parameters evaluated included visual analog scale pain scores at rest and while coughing, supplemental analgesic requirement, complications, hemodynamics, and respiratory parameters. Measurements were made at 2-hour intervals for 12 hours beginning at 10 minutes after endotracheal extubation. There was no statistically significant difference in visual analog scale scores and requirement of supplemental analgesia between the 2 groups. Cardiac index at 4 hours and 6 hours was significantly higher in the TEA group. Patients in the PVB group had significantly lower respiratory rates at 8, 10, and 12 hours. All other parameters were comparable. In 1 patient, the epidural space could not be catheterized. One patient in the TEA group had transient hypotension, and 1 patient complained of backache at the site of the epidural catheter insertion. CONCLUSION: PVB is as effective as TEA for postoperative analgesia after MIDCAB surgery. PVB is technically easier than TEA and may be safer than TEA because no complications were seen in the PVB group.

Pneumoencephalus and convulsions after ventriculoscopy: a potentially catastrophic complication.

A nine-year-old boy with hydrocephalus underwent ventriculoscopy under general anesthesia. After introduction of ventriculoscope the patient had sudden bradycardia, hypotension, and shrinkage of ipsilateral cerebral hemisphere. The ventriculostomy was abandoned. At the end of anesthesia and endotracheal extubation, the patient developed generalized convulsions. Reexploration of wound did not reveal anything significant; however, postoperative CT scan of head showed massive pneumoencephalus. The patients received elective ventilation of lungs for 24 hours and made complete recovery. The authors describe the reasons for these complications and further management.

Incidence of coronary artery disease in asymptomatic uncomplicated essential hypertensive patients--evaluation by treadmill exercise test.

Treadmill exercise test was performed in 100 patients, 50 were hypertensive who were not having any clinical or electrocardiographic manifestation of coronary artery disease and 50 were controls. The test was positive in 28% of hypertensive patients as compared to 6% in controls. This study therefore suggests that hypertension is frequently associated with asymptomatic coronary artery disease as compared to normotension and it is concluded that exercise electrocardiography test is a definite diagnostic tool in diagnosis of coronary artery disease not only in symptomatic but also in asymptomatic patients with one or more risk factors.

Use of traditional medical practitioners to delivery family planning services in Uttar Pradesh.

This pilot study conducted in Muzaffarnagar district in the state of Uttar Pradesh, India, demonstrates the potential for using traditional medical practitioners in the delivery of family planning services after a brief training period. Practitioners participated continuously in the program for an intervention period of two years during which their services were accepted by the community. The impact of their involvement was reflected in increased knowledge of permanent as well as reversible contraceptive methods and in higher contraceptive use rates, especially of reversible methods adopted by women younger than 25 years (from 8 percent to 37 percent), in the intervention villages, as compared with increased knowledge and use (from 13 percent to 25 percent) of permanent methods alone in the control villages.

Immune status of children suffering from minimal change nephrotic syndrome.

Twenty five children suffering from minimal change nephrotic syndrome were studied for immunological alterations at different stages of this disease i.e., onset, relapse and remission. Changes were found mainly at onset and during relapse in the form of altered helper and suppressor cell ratio, depressed delayed cutaneous hypersensitivity reaction, decreased S-IgG bearing lymphocytes with low serum IgG concentration, and increased S-IgM bearing lymphocytes with high serum IgM concentration. Majority of these parameters returned to normal values during remission. Serum IgE was found high at all stages of this. These alterations suggest defects in cell mediated immunity resulting in secretion of some substance which modifies the glomerular anionic charges.

An epidemiologic study of acute intermittent porphyria in Maheshwari community of Sri-Dungargarh municipal area.

Fifteen patients of acute intermittent porphyria (AIP) were detected out of 2500 persons of Maheshwari community surveyed. These 15 patients belonged to 11 families. The prevalence of AIP in Maheshwari population came out to be 1:640 which is considerably higher than the figures reported from many other areas of our country. AIP was more common in females than males, the ratio being 9:6. Maximum number of cases (6) were encountered in the age group of 11-20 years. AIP was observed to be inherited as an autosomal dominant disease in 3 families. Spontaneous mutation of gene may be considered to be the mode of inheritance in the remaining 8 families. The disease was found to be more common in Daga and Tapadia subcastes. There were 5 cases belonging to Daga subcaste and 3 belonging to Tapadia subcaste.

Carrier presentation: a possible strategy for selective in vivo amplification of specific T cells.

A strategy for selective in vivo amplification of specific T cells by carrier presentation is outlined here. This hypothesis addresses the possible physiological significance of expression of MHC II molecules by T cells, and attempts to highlight the possibility of harnessing the phenomenon for therapeutic use in medicine.