Association of interleukin-10 polymorphisms with chronic hepatitis C virus infection in a case-control study and its effect on the response to combined pegylated interferon/ribavirin therapy.

We conducted a case-control study involving 150 genotype 3 chronic hepatitis C virus (HCV) patients and 150 healthy controls to investigate the association of polymorphisms in the interleukin-10 (IL-10) gene with chronic HCV infection and the association of these polymorphic variants with the combination of pegylated interferon (Peg-IFN) and ribavirin therapy response. Our data revealed that the GG genotype of IL-10 -1082A/G exhibited significant association with genotype 3 chronic HCV infection compared to controls. Treatment response data also showed a significant increase in risk for the GG genotype of IL-10 -1082A/G in response-relapse patients or non-responder patients compared to sustained virological response patients. Further, a significant increase in risk was also revealed for the CC genotype of IL-10 -592A/C in response-relapse patients or non-responder patients compared to sustained virological response patients, suggesting a role of the GG genotype of IL-10 -1082A/G and CC genotype of IL-10 -592A/C in the treatment outcome of combined Peg-IFN/ribavirin therapy.

Alterations of p53 gene in gallbladder cancer patients of North India.

BACKGROUND: Mutations in p53 gene are found in a majority of human malignancies and usually occur in the exons 5, 6, 7 and 8. Mutated p53 protein is more stable and gets accumulated in the cells that induce the host to develop anti-p53 antibodies in sera of cancer patients. AIM: This study is aimed to observe the frequency and nature of mutations in exons 5-8 of p53 gene and to evaluate its correlation with prevalence of serum p53 antibodies in Indian patients with gallbladder cancer (GBC). METHODS: Mutation studies were done in cancer tissues obtained from 62 patients with proven GBC (40 cytologically proven cases and 22 resected gallbladder cancer tissues) by polymerase chain reaction (PCR), restriction fragment length analysis (RFLP) and single strand conformation polymorphism (SSCP). Presence of serum p53 antibodies was determined using highly specific enzyme linked immunosorbent assay (ELISA) kit in 50 patients with GBC and 30 patients of cholelithiasis. Clinicopathologic characteristics of these patients were given attention. RESULTS: Antibodies to p53 protein was present in the serum in 34% (17/50) of GBC patients and in 3.3% (1/30) patients with cholelithiasis (p < 0.018). RFLP failed to detect common mutations in the exons 5- 8 of the p53 gene in 62 samples. Using SSCP analysis we could detect frameshift mutation in p53 gene in 2 of 22 (9.1%) GBC cases. Mutated samples were sequenced and found to have insertion of adenine at codon 271 (GAG) in exon 8 region. CONCLUSION: Our results show that 1//3rd of the north Indian patients with GBC have antibodies to p53 protein. The commonest identifiable alteration in the p53 gene was a frameshift mutation at codon 271.

A school-based intervention of screening a movie to increase hepatitis B vaccination levels among students in Uttar Pradesh, India: impact on knowledge, awareness, attitudes and vaccination levels.

BACKGROUND: India is home to one in 14 of all chronic hepatitis B virus (HBV) cases, meaning that it is important to develop HBV interventions that are applicable in the Indian context. Vaccination is the foremost tool for interrupting the HBV infection cycle. HBV vaccination was not included in India's government-sponsored expanded immunisation program until 2011, and many children born earlier remain unvaccinated. This study sought to observe the impact of the HOPE Initiative's school-based intervention to increase vaccination coverage by increasing HBV awareness among students in Lucknow, Uttar Pradesh. METHODS: At 430 schools in the administrative areas within and surrounding Lucknow, students viewed an educational documentary film on HBV and completed two questionnaires, one immediately before the screening and the other six weeks later. Both questionnaires asked the same 14 questions, which were organized into five domains: knowledge of the magnitude of the problem of HBV; knowledge of modes of HBV transmission; knowledge of consequences of HBV infection; awareness of HBV; and attitudes regarding HBV. The baseline questionnaire also asked students whether they had been vaccinated against HBV. At two-year follow-up, researchers measured vaccination levels at a subset of 30 intervention schools and six non-intervention schools to further assess the impact of the intervention. RESULTS: Baseline questionnaires were completed by 11,250 students, and post-intervention questionnaires, by 9698 students. Scores for knowledge about the magnitude of the HBV problem improved from 41% at baseline to 74% at follow-up, and scores for knowledge about modes of transmission, from 38% to 75% (p < 0.05 for both). The baseline HBV vaccination level among students receiving the intervention was 21%. Two years after the intervention, 45% of students (N = 4284) reported being vaccinated at intervention schools compared to 22% (N = 1264) at non-intervention schools. CONCLUSIONS: The observed increases in HBV awareness, knowledge and vaccination levels in this study indicate that school-based interventions can be used to achieve higher vaccination coverage among Indian children. The documentary film was found to be an affordable tool for reaching large audiences. More studies are needed to validate the impact of this intervention and to explore its applicability to other social causes.

Identification of a New Genotype of African Swine Fever Virus in Domestic Pigs from Ethiopia.

African swine fever (ASF) is an important emerging transboundary animal disease (TAD), which currently has an impact on many countries in Africa, Eastern Europe, the Caucasus and the Russian Federation. The current situation in Europe shows the ability of the virus to rapidly spread, which stands to threaten the global swine industry. At present, there is no viable vaccine to minimize spread of the disease and stamping out is the main source of control. In February 2011, Ethiopia had reported its first suspected outbreaks of ASF. Genomic analyses of the collected ASF virus (ASFV) strains were undertaken using 23 tissue samples collected from domestic swine in Ethiopia from 2011 to 2014. The analysis of Ethiopian ASFVs partial p72 gene sequence showed the identification of a new genotype, genotype XXIII, that shares a common ancestor with genotypes IX and X, which comprise isolates circulating in Eastern African countries and the Republic of Congo. Analysis of the p54 gene also followed the p72 pattern and the deduced amino acid sequence of the central variable region (CVR) of the B602L gene showed novel tetramer repeats not previously characterized.

High rates of early HBeAg seroconversion and relapse in Indian patients of chronic hepatitis B treated with Lamivudine: results of an open labeled trial.

BACKGROUND: The use of Lamivudine in chronic hepatitis B (CHB) is well known, however the reported rate of HBeAg sero-conversion and its durability post-treatment have varied considerably. We undertook the present study to study the effect of Lamivudine on HBeAg loss and seroconversion rates in Indian patients of CHB in relation to frequency, predictors and durability. METHODS: We treated 60 patients of e antigen positive CHB (with active viral replication and ongoing necro-inflammatory activity) with Lamivudine. They were followed up by monthly aminotransferases, and 3 monthly HBeAg and anti-HBe. Those who attained HBeAg sero-conversion were advised to discontinue Lamivudine after 6 months and followed up every 3 months thereafter, to see for relapse. Treatment was given for maximum of 3 years if not sero-converted. RESULTS: The annual incremental loss of HBeAg in patients receiving Lamivudine was 25 (41.6%) at end of 1st year, 33 (55%) at 2nd year and 35 (58.3%) at 3rd year. The corresponding rates for full sero-conversion were 17/60 (28.6%), 22/60 (36.6%) and 24/60 (40%) in the 3 years. HBeAg loss correlated with increased pre-therapy ALT levels (p = 0.002) and decreased pretreatment HBV-DNA levels (p = 0.004). The presence of cirrhosis had no influence on the rate of HBeAg loss. Relapse occurred in 35% (7/20) post-treatment at median time of 6 months. CONCLUSION: Indian patients showed a higher rate of HBeAg sero-conversion in the first year of Lamivudine treatment. This correlated with baseline ALT and inversely with HBV-DNA levels. Relapse rate after treatment was high and occurred soon after stopping treatment.

Effects of locus coeruleus stimulation on neuronal activities of dorsal lateral geniculate nucleus and perigeniculate reticular nucleus of the rat.

In rats anesthetized with urethane, a stimulating electrode was introduced to the locus coeruleus by observing the antidromic field response to single shock stimulation of the dorsal pathway of noradrenergic axons. Effects of locus coeruleus stimulation were studied on activities of relay neurons and intrinsic interneurons of the dorsal lateral geniculate nucleus and on those of neurons in the perigeniculate reticular nucleus. The intrinsic interneurons and the perigeniculate reticular neurons are believed to exert inhibition upon the relay neurons. The relay neurons were activated by repetitive stimulation of locus coeruleus; spontaneous discharges were increased in rate and the threshold of response to single shock stimulation of the optic nerve was lowered. The activation was rarely seen in rats pretreated with alpha-methyl-p-tyrosine. Iontophoretic application of phentolamine, an alpha-blocker, effectively antagonized the activation, whereas an iontophoretic beta-blocker and cholinergic blockers were virtually ineffective. The activation of the relay neurons was suggested to be due to a direct action of noradrenaline, released by locus coeruleus stimulation. Locus coeruleus stimulation inhibited the interneurons and activated the perigeniculate reticular neurons; spontaneous or light-evoked discharges were suppressed in the interneurons and tonic discharges were elicited in the perigeniculate reticular neurons. These effects of locus coeruleus stimulation were mimicked by noradrenaline applied iontophoretically. Activation of the perigeniculate reticular neurons was antagonized by an iontophoretic alpha-blocker but not by a beta-blocker. Two special features emerge from the present results: (1) the locus coeruleus exerts different effects upon the two neuronal constituents of the dorsal lateral geniculate nucleus, excitation of the relay neurons and inhibition of the intrinsic interneurons; (2) a suggestion previously advocated that locus-coeruleus-induced excitation of the lateral geniculate relay neurons would be due to inhibition of inhibitory neurons (disinhibition) does not hold true, at least with respect to the perigeniculate reticular neurons; the latter neurons have been proved to exert a powerful inhibition upon the geniculate relay neurons and they are excited by stimulation of the locus coeruleus.

Ipsilateral motor control of the forelimb in the congenitally acallosal mouse.

The corticospinal projections of mice with callosal agenesis were investigated using electro-physiological and horseradish peroxidase techniques. In the normal mouse, intracortical stimulation of the motor area with the microelectrode resulted in contralateral contraction of the forelimb, whereas ipsilateral contraction was observed in the acallosal mouse. Hence, mice with congenital absence of the corpus callosum show physiologically ipsilateral motor control. Furthermore, latency of the forelimb contraction elicited by electrical stimulation of the acallosal mouse was slightly shorter than that of the normal mouse. On the other hand, it was confirmed by a horseradish peroxidase technique that the corticospinal tract and peripheral motor neurons in the spinal cord of the acallosal mouse were identical to those of the normal mouse.

Suppression of hyperemia after brain ischemia by L-threo-3,4-dihydroxyphenylserine.

Although several studies have shown that L-threo3,4-dihydroxyphenylserine (DOPS) may provide a neuroprotective effect against ischemic brain damage, its protective mechanism is not fully understood. Glutamate release and hippocampal blood flow in ischemia with administration of DOPS were investigated to elucidate the neuroprotective mechanism of DOPS. Pre- (but not post-) ischemic administration of DOPS rescued 73% of hippocampal CA1 neurons (p < 0.001, compared with ischemia only) 1 week after transient global ischemia in gerbils. While glutamate release induced by ischemia was not affected, the increase of hippocampal blood flow during reperfusion was significantly suppressed by DOPS. These results demonstrate that DOPS may prevent reperfusion injury by suppression of hyperemia after ischemia, resulting in neuroprotection.

Clostridium perfringens epsilon toxin causes excessive release of glutamate in the mouse hippocampus.

The mechanism of neurotoxicity of Clostridium perfringens epsilon toxin to the mouse brain was investigated. Intravenous injection in mice with the toxin caused seizure and excited hippocampal neurons. Microdialysis revealed that epsilon toxin induced excessive glutamate release in the hippocampus. Both the seizure and glutamate release were attenuated by prior injection with riluzole, an inhibitor of pre-synaptic glutamate release, suggesting that this toxin enhances glutamate efflux, leading to seizure and hippocampal neuronal damage.

Depression of long term potentiation in gerbil hippocampus following postischemic hypothermia.

To investigate the mechanism of chronic cell death following postischemic hypothermia, the change of N-methyl-D-aspartate receptor (NMDAR) were examined by immunohistochemistry of NMDAR1 and long-term potentiation (LTP) in the CA1 subfield of the gerbil hippocampus. At 1 week following postischemic hypothermia (32 degrees Cx4 h), all CA1 neurons survived; however, immunoreactivity of NMDAR1 increased in neuronal perikarya whereas decreased in dendrites in the CA1 neurons. The abnormality was still observed in remaining CA1 neurons at 1 month after hypothermia. LTP was also significantly depressed at 1 week after hypothermia. These results suggest that some abnormalities in the glutamate receptor may be caused by ischemia; such abnormality would persist in spite of hypothermia treatment, resulting in the depression of LTP.

Effect of embryonic hippocampal transplantation in amygdaloid kindled rat.

Embryonic neural tissue was transplanted into previously kindled rats. A thirteen- to fourteen-day embryonic hippocampal cell suspension was grafted in the stratum oriens near the CA2 area of the hippocampus. Almost 80% of the animals had a good recovery and became seizure-free. Injection of neocortical cells or saline did not show any positive effect on the kindling susceptibility. Although 20 day embryonic cell transplantation was also effective, the effect did not last as long as the 13- to 14-day embryonic transplantation. These observations open the possibility that the neural grafts may be used for therapy of medically intractable epilepsies.

Effect of ceruletide on epileptogenesis in amygdaloid kindled rats.

The inhibitory effects of ceruletide (CLT), a cholecystokinin-8 (CCK)-like peptide, were investigated in the epileptogenesis in the amygdaloid kindled rats. Lower doses of CLT (20-80 micrograms/kg) inhibited the progression of kindling process. After acquiring C5 stage, a higher dose (160 micrograms/kg) was required to suppress the seizure susceptibility. These results, in light of several previous studies showing no serious side effects, suggest that CLT might be useful as an anti-epileptogenic agent for clinical usage.

In vivo microdialysis of amino acid neurotransmitters in the hippocampus in amygdaloid kindled rat.

Extracellular concentrations of gamma-aminobutyric acid (GABA), glutamate (Glu) and aspartate (Asp) were determined by microdialysis in rat hippocampus during various amygdaloid kindled stages. The values of GABA and Glu were increased 3-4 times in C2-C3 stages in comparison with the values in control animals. After reaching the C5 stages, these values were increased 3-7 times. However, the concentration of Asp decreased depending on the kindling stage, reaching the lowest value of 33% in comparison with the normal value. The observed changes may be related to kindling induced seizures.

Ceruletide suppresses rotational behavior in lesioned rats via CCKA receptors.

The effects and pharmacological mechanisms underlying the inhibiting effect of ceruletide, a cholecystokinin (CCK)-related peptide, on apomorphine-induced turning behavior in 6-hydroxydopamine lesioned rats were investigated. For this purpose, selective CCKA and CCKB receptor antagonists were used. Ceruletide (50-400 micrograms/kg, s.c.) dose dependently suppressed apomorphine-induced rotational behavior. The antiapomorphine effect of ceruletide was reversed by the selective CCKA receptor antagonist, devazepide, but not by the selective CCKB receptor antagonist, L-365,260. This suggests that the suppression ceruletide exerts on hyperactive nigrostriatal dopamine neurons is mediated by CCKA receptors.

Long-term activation of the glutamatergic system associated with N-methyl-D-aspartate receptors after postischemic hypothermia in gerbils.

OBJECTIVE: The objective of this study was to investigate whether hypothermia would suppress secondary damage in the chronic postischemic stage, in terms of glutamate excitotoxicity. METHODS: Gerbils underwent 5 minutes of ischemia via bilateral common carotid artery occlusion. Seven groups were studied, as follows: 1) ischemia without treatment group; 2) intraischemic hypothermia group; 3) postischemic hypothermia group (32 degrees C for 4 h); 4) MK-801 treatment group (2 mg/kg, every other day for 1 mo); 5) postischemic hypothermia with MK-801 treatment for 1 week group (2 mg/kg, every other day); 6) postischemic hypothermia with MK-801 treatment for 1 month group (2 mg/kg, every other day); and 7) sham-treated control group. One month after ischemia, histological changes in hippocampal CA1 neurons (assessed using hematoxylin and eosin staining) and memory function (assessed using an eight-arm radial maze) were studied. Extracellular glutamate concentrations were monitored by microdialysis during ischemia and hypothermia. Staining of microglia was performed 1 week and 1 month after ischemia. RESULTS: MK-801 alone, postischemic hypothermia alone, and postischemic hypothermia with MK-801 treatment for 1 week failed to prevent ischemic neuronal damage and memory function decreases 1 month after the insult (P < 0.05 versus control). However, the postischemic hypothermia with MK-801 treatment for 1 month group exhibited significant protective effects (not significant [P > 0.05] compared with the control group). Extracellular glutamate levels for the intraischemic hypothermia group were significantly low, compared with the postischemic hypothermia group. There was no microglial activation in the postischemic hypothermia at 1 week and 1 month after ischemia groups. CONCLUSION: Postischemic hypothermia and long-term intermittent administration of MK-801 demonstrated significant neuronal protection, indicating that long-term glutamatergic activation, with changes in N-methyl-D-aspartate receptors, plays a role in neuronal damage in the chronic postischemic stage.

Morphological changes in the hippocampus in amygdaloid kindled mouse.

To clarify the origin and maintenance of epileptogenesis, morphological changes in the hippocampus of amygdaloid-kindled mice were analyzed at different stages of kindling. The granule cell size in dentate gyrus and the pyramidal cell size in CA1 were clearly decreased depending on seizure stage. The cell size in CA2 was increased and density in dentate gyrus and CA2 was reduced, significantly. The morphological changes in hippocampus associated with kindling must be closely related to the acquisition and the maintenance of epileptogenesis. The results support the hypothesis that seizure-induced damage of neurons may lead to formation of new synaptic connections that produce abnormal hyperexitability and result in seizures.

Forelimb muscle contraction induced by cerebral cortical stimulation after callosotomy: a myographic study in the mouse.

The effect of callosotomy upon motor control by the cerebral cortex on the forelimb was examined in the mouse. On the 10th day after callosotomy in the rostral or caudal part of the corpus callosum, the forelimb area of the cerebral motor cortex was stimulated intracortically with a microelectrode and muscle contraction of the forelimb was recorded by electromyography. Muscle contraction was observed in the contralateral forelimb in the mice of which callosal fibers were cut in the caudal part of the corpus callosum including the splenium corporis callosi as well as in the normal mice. On the other hand, muscle contraction was observed in the ipsilateral forelimb in the mice of which callosal fibers were incised in the rostral part of the corpus callosum including the genu, rostrum and trunk. The latency of the muscle contraction was about 0.5 msec in the both groups of the callosotomized mice, as well as in the normal mice.

Changes in the spine density on apical dendrites of pyramidal neurons in the motor area of the cerebral cortex after callosotomy: a study by a modified Golgi-Cox method in the mouse.

Changes in the number of spines on apical dendrites of pyramidal neurons in layers III and V of the motor area of the cerebral cortex were examined in the young adult mice (30-60 days old) by a modified Golgi-Cox method and laser scanning microscopy on the 1st, 6th, 10th and 12th days after callosotomy. The anterior part of the callosal body, including rostrum, genu and truncus corporis callosi, was sectioned with a razor blade. The numbers of spines on apical dendrites and their oblique branches of pyramidal neurons were counted at the level of layer II for layer III pyramidal neurons and at the level of layer III for layer V pyramidal neurons. The density of spines was increased on all these dendritic parts during 10 days after callosotomy; the diameters of spine stems and dendrites were also increased. These changes, however, appeared to be transient as they were decreased almost to normal level on the 12th day after callosotomy.

[Effects of dithizone on the electroencephalogram recorded from the mouse hippocampus in vivo].

In the present study the author examined the effects of dithizone on hippocampal and cortical EEG by power spectral analysis in the moving mouse. Following results were obtained. Administration of dithizone 100 mg/kg i. p. produced almost loss of electrical activities on EEG which began 409 sec after injection and lasted approximately up to 706 sec. In recovery period waveform showed shift to slower frequencies apparent by 60 min. Heart rate decreases were seen between 5 and 20 min after 100 mg/kg i. p. injection. Dithizone produced dose-dependent changes in hippocampal and heart rate activities. Abolished EEG by dithizone administration were immediately recovered by zinc-acetate application. Injection of vehicle had no significant effect on hippocampal and cortical EEG.

[Effects of chelating reagents on the hippocampal EEG and histochemical Timm staining pattern in a mouse brain].

A series of experiments we examined in detail the effects of four chelating reagents, dithizone, DEDTC, oxine and EDTA, upon (1) Timm's staining in the hippocampal formation, (2) brain electrical activity recorded from hippocampal and cortical electrodes, (3) recovery effect with zinc acetate or zinc sulfate on abolished brain electrical activity, (4) open field activity, (5) toxicity, (6) and heart rate. Dithizone, DEDTC and oxine influenced all measures, and the degree of effect varied directory with dose. But EDTA has not any significant effect on biological system. Resulting data were summarized in Table 4.