Lentiviral Vectors Displaying Modified Measles Virus gp Overcome Pre-existing Immunity in In Vivo-like Transduction of Human T and B Cells

Gene transfer into quiescent T and B cells is important for gene therapy and immunotherapy approaches. Previously, we generated lentiviral vectors (LVs) pseudotyped with Edmonston (Ed) measles virus (MV) hemagglutinin (H) and fusion (F) glycoproteins (H/F-LVs), which allowed efficient transduction of quiescent human T and B cells. However, a major obstacle in the use of H/F-LVs in vivo is that most of the human population is vaccinated against measles. As the MV humoral immune response is exclusively directed against the H protein of MV, we mutated the two dominant epitopes in H, Noose, and NE. LVs pseudotyped with these mutant H-glycoproteins escaped inactivation by monoclonal antibodies (mAbs) but were still neutralized by human serum. Consequently, we took advantage of newly emerged MV-D genotypes that were less sensitive to MV vaccination due to a different glycosylation pattern. The mutation responsible was introduced into the H/F-LVs, already mutated for Noose and NE epitopes. We found that these mutant H/F-LVs could efficiently transduce quiescent lymphocytes in the presence of high concentrations of MV antibody-positive human serum. Finally, upon incubation with total blood, mimicking the in vivo situation, the mutant H/F-LVs escaped MV antibody neutralization, where the original H/F-LVs failed. Thus, these novel H/F-LVs offer perspectives for in vivo lymphocyte-based gene therapy and immunotherapy.     

Fenamate-induced enhancement of heterologously expressed HERG currents in Xenopus oocytes

In the present study, the effects of intra-nucleus accumbens injection of L-arginine, a nitric oxide (NO) precursor, and N(G)-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase (NOS) inhibitor, on morphine-induced conditioned place preference in male Wistar rats were investigated. Our data showed that subcutaneous (s.c.) injection of morphine sulphate (0.5-10 mg/kg) significantly increased the time spent in the drug-paired compartment in a dose-dependent manner. Intra-accumbens administration of L-arginine (0.03 and 0.05 microg/rat) with an ineffective dose of morphine (0.5 mg/kg) elicited significant conditioned place preference, while intra-accumbens administration of L-NAME (0.3, 0.1 and 1 microg/rat) decreased the acquisition of conditioned place preference induced by morphine (7.5 mg/kg). The response to different doses of L-arginine was decreased by L-NAME (0.03 microg/rat). L-Arginine and L-NAME by themselves did not elicit any effect on place conditioning. Intra-accumbens administration of L-arginine but not L-NAME significantly decreased the expression of morphine (7.5 mg/kg)-induced place preference. The attenuation of already established morphine-induced place preference on the test day by L-arginine was inhibited by L-NAME. The results indicate that NO may be involved in the acquisition and expression of morphine-induced place preference.     

Modulation of tricarboxylic acid cycle dehydrogenases during hepatocarcinogenesis induced by hexachlorocyclohexane in mice

The sequential distribution of key tricarboxylic acid (TCA) cycle enzymes have been investigated during hexachlorocyclohexane (HCH)-induced hepatocarcinogenesis in Swiss mice. Animals were continuously exposed to HCH (500 ppm) for 2, 4, and 6 months until liver tumor developed. The activity of TCA cycle enzymes such as isocitrate dehydrogenase (ICDH), succinate dehydrogenase (SDH), and malate dehydrogenase (MDH) have been studied. The activity of all the enzymes declined after 2 months of exposure of HCH in the liver. The neoplastic nodules and tumors developed after an exposure of HCH for 4 and 6 months, respectively. Neoplastic nodule and tumor showed wide variations in the activity and distribution of TCA cycle enzymes. The decreasing pattern in the activity of enzymes persisted in the non-neoplastic and non-tumor regions of the liver except SDH. However, the cells in nodular area and tumor showed intense enzymatic activities at cellular level. In the nodular region SDH activity declined prominently, whereas the non-nodular area showed positive reaction. Conspicuously, the tumor showed islands of positive and negative zones for TCA cycle dehydrogenases. The significance and relevance of such a distribution pattern still remains a mystery. The results are discussed in the light of HCH-induced toxicity on energy metabolism in exposed animals and possible role of such enzymes in the tumor formation.     

Dexmedetomidine used to provide hypotensive anesthesia during middle ear surgery

This study was carried out to assess the hypotensive effect of low dose dexmedetomidine (DEX) infusion during middle ear surgery. 42 ASA grades I and II patients of either sex aged 18–45 years undergoing elective middle ear surgery were randomly divided into two groups of 21 each. Group I received placebo bolus and infusion of saline at a rate similar to DEX in Group II. Group II received 10–15 min prior to induction of anesthesia 1 µg/kg IV bolus DEX diluted in 10 ml of normal saline over 10 min. Immediately thereafter an infusion of 0.4 µg/kg/hr of DEX commenced. Standard anesthetic technique was used. Halothane was titrated to achieve a mean arterial pressure 30% below the control value (value taken just after premedication). We observed that a statistically significant reduction in the percentage of halothane required to reduce MAP 30% below control value occurred in patients receiving DEX infusion (1.3 ± 0.4%) in comparison to those receiving placebo (3.1 ± 0.3%). Patients receiving DEX infusion had a better surgical field. The mean awakening time was significantly reduced in patients of Group II (9.1 ± 2.7 min) when compared to patients of Group I (12.8 ± 2.2 min). We conclude that DEX can be safely used to provide hypotensive anesthesia during middle ear surgery.     

Prevalence,disease associations and risk factors for colonization with intestinal spirochaetes (Brachyspira spp.) in flocks of laying hens in north-eastern Italy

The present study investigated the occurrence of anaerobic intestinal spirochaetes of the genus Brachyspira in laying hen flocks in Treviso province, north-eastern Italy, with respect to prevalence, spirochaete species present, disease associations and risk factors for colonization. A total of 450 faecal samples from 45 sheds on 29 laying hen farms were cultured for intestinal spirochaetes. Nineteen sheds on 12 farms contained chickens with symptoms consistent with avian intestinal spirochaetosis, including reduced egg production, wet litter and/or pasty vents. Spirochaetes were isolated from 157 (34.8%) samples from 21 (72.4%) farms, and from 32 (71.1%) sheds. From these positive samples, 189 spirochaetal isolates were speciated using three polymerase chain reaction assays and a restriction fragment polymorphism analysis of 16S rDNA polymerase chain reaction products. Overall, 52 (27.5%) isolates were identified as pathogenic Brachyspira intermedia, 26 (13.8%) as pathogenic Brachyspira pilosicoli, 93 (49.7%) as non-pathogenic (Brachyspira innocens/Brachyspira murdochii), and 18 (9.6%) were unidentified. Faeces from 14 sheds (31%) on 10 farms (34.5%) contained B. intermedia and/or B. pilosicoli, and disease consistent with avian intestinal spirochaetosis was observed in nine of these sheds on seven farms. There was a significant association (P=0.042) between the presence of spirochaetes and using deep pits rather than conveyor belts for manure disposal. Sheds housing chickens >40 weeks of age were significantly more likely to contain spirochaetes (P=0.048) and pathogenic species (P=007) than sheds housing younger chickens. A significant association (P=0.02) was found between infection with pathogenic spirochaetes and reduced egg production.     

Knowledge and attitude towards tuberculosis in a slum community of Delhi

A cross-sectional study was conducted among 208 adults (114 males and 94 females) aged 16-70 years, selected by systematic random sampling method in Lok Nayak Colony, Delhi to assess the knowledge and attitude towards tuberculosis (TB). Literacy rate was 28.4%. Only 174 (83.6%) heard of tuberculosis mainly from neighbours (64.9%) and friends (62.1%). Only 2.3% knew that TB was caused by a germ. Literates were more aware than illiterates regarding some signs and symptoms of TB i.e breathlessness (p=0.002), low grade fever (p=0.02), loss of appetite (p<0.001) and factors favouring TB e.g. overcrowding (56.4%) and poor diet (45.4%). Only 12.6% knew about the duration of treatment for 6-8 months and 1.7% knew about preventive role of BCG. Tendency to discriminate TB patients was evident from the findings e.g 71% respondents agreed upon isolating TB patients from the family, 74.1% on avoiding the patient in food sharing, on quitting job by the patient (33%), prohibiting marriage of the patient (27.6%), shunning him from attending social functions (18%), etc. Extensive health education directing towards attitudinal change by community involvement is needed to create awareness and remove myths about TB in such colonies.