trans-4,4'-Dihydroxystilbene (DHS) protects PC12 cells from oxidative damage but induces reactive oxygen species-mediated apoptosis in SHSY-5Y cell line.

Polyphenols can exert both, antioxidant and pro-oxidant properties, depending on cell types as well as their concentrations. Hence, it was of interest to examine if the naturally occurring resveratrol analog, trans-4,4'-dihydroxystilbene (DHS) also exert both these activities in a biphasic or cell-specific manner. In this study, we established the cytoprotective action of DHS against hydrogen peroxide (H2O2)-induced apoptotic death of the PC12 cells. DHS reduced mitochondrial membrane permeabilization and deactivated reactive oxygen species (ROS)-mediated caspase-3 activation in the H2O2-treated PC12 cells. However, it induced apoptosis in the human neuroblastoma SHSY-5Y cell line by destabilizing mitochondrial membrane, augmenting ROS and activating caspapse-3. DHS showed better activity than resveratrol in both the chosen models.

Association of Oxidative Stress and Obesity with Insulin Resistance in Type 2 Diabetes Mellitus.

Oxidative stress occurs due to delicate imbalance between pro-oxidant and anti oxidant forces in our system. It has been found to be associated with many morbidities but its association with obesity and insulin resistance is still controversial. Here in our study we examined 167 patients of recent onset type 2 diabetes mellitus and 60 age sex matched non-diabetic control. Body Mass Index (BMI), abdominal circumference, fasting blood glucose, serum insulin and plasma Malondealdehyde (MDA, marker for oxidative stress) were measured in them. On the basis of BMI, subjects were divided into obese (BMI≥25) and non obese (BMI<25) groups. Insulin resistance scores were calculated by Homeostatic Model Assessment-Insulin Resistance (HOMA-IR) method. Physical parameters (BMI, abdominal circumference) as well as levels of insulin and MDA were found to be significantly higher in subjects with diabetes than their non diabetic controls. The said parameters also showed significant difference in obese and non-obese sub groups. Insulin resistance score showed positive correlation with BMI, abdominal circumference, and plasma MDA, strength of association being highest with abdominal circumference. Plasma MDA was found to have positive correlation with physical parameters. Study concludes that, obesity mainly central type may predispose to insulin resistance and oxidative stress may be a crucial factor in its pathogenesis. Thus, oxidative stress may be the connecting link between obesity and type 2 diabetes mellitus, two on going global epidemics.

Stress inducible heat shock protein 70: a potent molecular and toxicological signature in arsenic exposed broiler chickens.

This communication reports about heat shock protein response after arsenic exposure in broiler chickens in vivo and in vitro both. Splenocytes harvested in presence of sodium arsenite expressed Heat shock protein 70 (HSP 70) which could be identified on the basis of relative migration pattern and western blot analysis. Serum levels of HSP 70 in broiler chicken also increased after continuous feeding of sodium arsenite in drinking water. This particular observation may be attributed towards systematic inflammation, oxidative stress and hepatocellular injury. In vitro relative quantification of transcription level of HSP 70 revealed that splenocytes harvested in presence of sodium arsenite expressed (final concentration 3 and 7 µM/ml) more HSP 70 in comparison to cells harvested without sodium arsenite and the values were statistically significant (P < 0.001) when compared to untreated control. Collectively this result indicated that, HSP 70 level increased both in vivo and in vitro trials and may be used as potential molecular and toxicological biomarker.

Mitochondrial medicine.

Mitochondrial diseases are extremely heterogeneous multisystem disorders predominantly affecting tissues or organs with high oxygen consumption like skeletal muscles, brain, endocrine glands, myocardium, eyes, ears, intestines, liver, kidneys, and bone marrow. Although various clinical syndromes have been described, they frequently overlap and there is no diagnostic gold standard to identify all. It is difficult to chart the future of an affected individual as also to predict the response to treatment which is mostly supportive and symptomatic. The rapidly increasing understanding of the pathophysiologic background of mitochondrial disorders may facilitate diagnostic approach and open perspectives to curative therapies. With the coming of age for mitochondrial medicine, it is now appropriate that physicians keep themselves well-acquainted with the recent developments in this expanding field of biomedical research.

Familial hypoparathyroidism.

A family with hereditary (familial) hypoparathyroidism is reported where three of the total four siblings were affected and each presented with different manifestation-one brother with refractory epilepsy since early childhood, another brother with unilateral extrapyramidal features in adult life, and their only sister having recurrent attacks of tingling and numbness due to hypocalcemia since 12 years of age.

Plasma levels of lipid peroxides in patients with Parkinson's disease.

Oxidative stress is implicated as a major factor for nigral neuronal cell death. Metabolic failure in antioxidant mechanisms could hypothetically facilitate the chemical processes that lead to lipid peroxidation. To elucidate whether elevated lipid peroxidation rates might increase risk of developing Parkinson's disease (PD), the Authors determined plasma levels of malondialdehyde (MDA) in 80 PD patients and 80 controls. There was a significant difference between the plasma MDA levels of PD patients and controls (7.48 +/- 1.55 vs 5.1 +/- 1.26 nmol/ml). Plasma MDA levels were inversely related to the age of the PD patients (r = -0.46; p < 0.01) and age of onset but in the control group, no such correlation was observed between the plasma MDA and age. However, there was no significant correlation between plasma MDA levels and the duration of disease, Hoehn and Yahr stages and the Unified Parkinson's Disease Rating Scale (UPDRS). Thus, the results suggest that high plasma lipid peroxidation rates might contribute as a risk factor for PD in West Bengal.

Profile of tuberculous meningitis with or without HIV infection and the predicators of adverse outcome.

OBJECTIVES: To compare the clinical, radiological and cerebrospinal fluid (CSF) findings, at hospital admission, among adult patients with tuberculous meningitis (TBM) with or without HIV infection and to identify the factors that predict adverse outcome at six months. METHODS: A total of 82 adult patients with TBM were included (40 HIV-positive and 42 HIV-negative). Several clinical (duration of illness, Glasgow Coma Scale score, presence of high temperature, headache, cranial nerve or sphincter abnormality, seizures and endocrine dysfunction), radiological (presence of hydrocephalus, cerebral infarction and oedema, meningeal enhancement, granuloma) and cerebrospinal fluid parameters (glucose, protein, lactate, lymphocytes, neutrophils and adenosine deaminase values) were recorded along with CD4 count in the peripheral blood. Statistical analysis was performed using the chi-square test. Individual variables were evaluated as prognostic factors for adverse outcome in both groups by calculating the relative risk of association for each. RESULTS: Temperature more than 38.33 degrees C was more common in the HIV-negative group while seizures, hydrocephalus, cerebral infarction and low CD4 count occurred significantly more commonly in the HIV-positive group. Hydrocephalus had strong association with severe neurological deficit and seizure with death in both the groups. CONCLUSION: Several clinical and laboratory features of TBM in patients who are HIV-positive are distinctly different from those without HIV infection; some of these have an association with the probability of adverse outcome.

Massive haemothorax: a presentation of pulmonary arteriovenous malformation.

Arteriovenous malformations of the lung are relatively uncommon lesions with varied clinical presentation. Nearly half of these are associated with Osler-Rendu-Weber disease. Magnetic resonance angiography is an accurate and non-invasive diagnostic modality. We report a case of a 56-year-old male who had massive haemothorax due to rupture of a pulmonary arteriovenous malformation arising from the right interlobar artery.

Gene induction by interferons: functional complementation between trans-acting factors induced by alpha interferon and gamma interferon.

HeLaM is a variant cell line in which the transcriptional induction of many genes by alpha interferon has special characteristics (Tiwari et al., Mol. Cell. Biol. 8:4289-4294, 1988). The same characteristics were also displayed for induced transcription of a permanently transfected chimeric gene containing the interferon-stimulated response element of gene 561. For understanding the molecular basis of the special requirements of HeLaM cells, an analysis of the interferon-stimulated gene factors (ISGF) was undertaken. By using gel shift assays, it was shown that the activation of ISGF3 by alpha interferon treatment of HeLaM cells had characteristics identical to those of induced transcription: inhibition by 2-aminopurine and the need for ongoing protein synthesis which was obviated by pretreating the cells with gamma interferon. Upon mixing in vitro the cytoplasmic fraction of gamma interferon-treated HeLaM cells with that of cells treated with alpha interferon and cycloheximide, active ISGF3 was reconstituted, presumably through complementation of two components, ISGF3 gamma and ISGF3 alpha, present in the two respective fractions. Because, unlike other cells, untreated HeLaM cells did not contain detectable levels of either component, we could induce them individually and study their independent properties. Induction of ISGF3 gamma but not of ISGF3 alpha needed ongoing protein synthesis and was blocked by 2-aminopurine. Once induced, ISGF3 gamma was active for 24 h and was present in both the nuclear and cytoplasmic fractions. Activated ISGF3 alpha, on the other hand, did not translocate to the nucleus in the absence of ISGF3 gamma, and in the cytoplasm its activity decayed within 2 h of its activation. In reference to our working model, all of the above observations indicate that ISGF3 gamma is the product of signal 1 and ISGF3 alpha is the product of signal 2.

Inhibition of double-stranded RNA- and tumor necrosis factor alpha-mediated apoptosis by tetratricopeptide repeat protein and cochaperone P58(IPK).

P58(IPK) is a tetratricopeptide repeat-containing cochaperone that is involved in stress-activated cellular pathways and that inhibits the activity of protein kinase PKR, a primary mediator of the antiviral and antiproliferative properties of interferon. To gain better insight into the molecular actions of P58(IPK), we generated NIH 3T3 cell lines expressing either wild-type P58(IPK) or a P58(IPK) deletion mutant, DeltaTPR6, that does not bind to or inhibit PKR. When treated with double-stranded RNA (dsRNA), DeltaTPR6-expressing cells exhibited a significant increase in eukaryotic initiation factor 2alpha phosphorylation and NF-kappaB activation, indicating a functional PKR. In contrast, both of these PKR-dependent events were blocked by the overexpression of wild-type P58(IPK). In addition, the P58(IPK) cell line, but not the DeltaTPR6 cell line, was resistant to dsRNA-induced apoptosis. Together, these findings demonstrate that P58(IPK) regulates dsRNA signaling pathways by inhibiting multiple PKR-dependent functions. In contrast, both the P58(IPK) and DeltaTPR6 cell lines were resistant to tumor necrosis factor alpha-induced apoptosis, suggesting that P58(IPK) may function as a more general suppressor of programmed cell death independently of its PKR-inhibitory properties. In accordance with this hypothesis, although PKR remained active in DeltaTPR6-expressing cells, the DeltaTPR6 cell line displayed a transformed phenotype and was tumorigenic in nude mice. Thus, the antiapoptotic function of P58(IPK) may be an important factor in its ability to malignantly transform cells.

Healing property of the Piper betel phenol, allylpyrocatechol against indomethacin-induced stomach ulceration and mechanism of action.

AIM: To evaluate the protective activity of allylpyrocatechol (APC), the major antioxidant constituent of Piper betel, against the indomethacin-induced stomach ulceration in the rat model and correlates with its antioxidative and mucin protecting properties. METHODS: Male Sprague-Dawley rats were divided into five groups. Normal control rats (group I) were given the vehicle oral dose of gum acacia in distilled water (1 mL per rat); ulcerated control and treated rats (groups II-V) were given a single dose of indomethacin (30 mg/kg body wt.); group II rats were sacrificed 4 h after indomethacin administration; groups III-V rats were given the vehicle (1 mL per rat) or APC (2 mg/kg body wt.) or misoprostol (1.43 mug/kg body wt.) once daily by oral intubation for 7 d starting from 4 h after the indomethacin administration. After 7 d, the stomach tissues were excised for histological examination and biochemical analysis. RESULTS: Treatment with APC (2 mg/kg body wt per day) and misoprostol (1.43 mug/kg body wt per day) for 7 d could effectively heal the stomach ulceration as revealed from the ulcer index and histopathological studies. Compared to the zero day ulcerated group, treatment with APC and misoprostol reduced the ulcer index by 93.4% and 85.4% respectively (P < 0.05). Both APC and misoprostol accelerated ulcer healing observed in natural recovery (P < 0.05), their respective healing capacities not being significantly different. The healing capacities of APC and misoprostol could be attributed to their antioxidant activity as well as the ability to enhance the mucin content of the gastric tissues. Compared to the ulcerated untreated rats, those treated with APC and misoprostol showed near normal MDA levels, while the protein levels were 86% and 78% of the normal value respectively (P < 0.05). Likewise, both APC and misoprostol increased the SOD, catalase, and mucin levels significantly (P < 0.05), the effect of APC being better. CONCLUSION: APC can protect indomethacin-induced gastric ulceration due to its antioxidative and mucin protecting properties.

Primary duodenal lymphoma producing obstructive jaundice.

Primary lymphoma of the duodenum presenting with obstructive jaundice is a rare entity. We report a case of primary non-Hodgkin's lymphoma of the duodenum producing obstructive jaundice in a middle aged lady, where the concentric thickening of the duodenal wall also gave rise to symptomatic partial high small bowel obstruction in due course. Guided aspiration and flowcytometry established a diagnosis of diffuse large B-cell lymphoma.

Sequence analysis of the non-structural 3A and 3C protein-coding regions of foot-and-mouth disease virus serotype Asia1 field isolates from an endemic country.

A total of 18 foot-and-mouth disease virus (FMDV) serotype Asia1 field isolates belonging to two different lineages (including the divergent group) as delineated earlier in VP1-based phylogeny were sequenced in the non-structural 3A and 3C protein-coding regions. The phylogenetic trees representing the regions coding for the non-structural proteins were very similar to that of the structural VP1 protein-coding region. Phylogenetic comparison at 3C region revealed clustering of Asia1 viruses with the isolates of serotypes O, A and C in the previously identified clade. Comparison of amino acid sequences identified lineage-specific signature residues in both the non-structural proteins. Overall analysis of the amino acid substitutions revealed that the 3A coding region was more prone to amino acid alterations than 3C region.

Analysis of the matrix protein gene sequence of the Asian lineage of peste-des-petits ruminants vaccine virus.

The M gene nucleotide sequence of an Indian peste-des-petits ruminants (PPRV) vaccine virus ("PPRV Sungri/96") belonging to Asian lineage was determined. The gene is 1476 nucleotides long with a single open reading frame (ORF). The nucleotide and predicted amino acid sequence was compared with the homologous region of the African Lineage Vaccine virus "PPRV/Nigeria/75/1". The nucleotide sequence of the "PPRV Sungri/96" was 86% identical to that of "PPRV/Nigeria/75/1", while a homology of 93% and 95% could be observed in the ORF and amino acids level, respectively. The M gene encodes a protein of 335 amino acids, with a predicted molecular weight (MW) of 37.8 kDa. The ORF is flanked by a 3' untranslated region of 436 nucleotides and a high level of sequence divergence (approximately 30%) could be observed in this region between the vaccine viruses of Asian and African lineages. A high degree of conservation of several amino acids of this protein observed previously was also confirmed in this study.

Comparative nucleotide sequence analysis of the phosphoprotein gene of peste des petits ruminants vaccine virus of Indian origin.

The nucleotide sequences of the phosphoprotein (P) gene of peste des petits ruminants (PPRV) vaccine virus (PPRV Sungri/96) belongs to Asian lineage have been determined and the deduced amino acid sequences were compared with another vaccine strain PPRV/Nigeria75/1 and with those of the other morbilliviruses. The 1652 nucleotides of the P gene encode a phosphoprotein of 509 amino acid residues (from nucleotide numbers 60 to 1587), which is 91% identical to that of PPRV/Nigeria75/1. The C protein consists of 177 amino acid residues and is 91% identical with that of PPRV/Nigeria75/1. The conserved mRNA editing site (5'TTAAAAGGGCACAG) was present at positions 742-756 in the P gene, which is conserved in all other morbilliviruses. The CTT trinucleotide sequence is present at the N/P and P/M intergenic region, which is totally conserved in morbilliviruses. This will be the third sequence for the P gene of PPRV since that of the vaccine strain and a wild-type Turkish isolate has been published already.

Sequence analysis of VP7 gene of Indian bluetongue virus serotype-23 shows its close phylogenetic relationship to Australian and Chinese serotypes.

Bluetongue, an arthropod borne viral disease of wild and domestic ruminants, causes heavy economic losses throughout the world. In the present study, full-length VP7 gene of Indian bluetongue virus (BTV) serotype 23 was sequenced and compared with prototype strains of BTV reported from different countries. Nucleotide sequence analysis of VP7 gene revealed Indian BTV serotype 23 to have 1154 nucleotides with the deletion of two nucleotides at 3' non-coding region and a unique amino acid change 211S-N. The Indian virus also demonstrated a maximum similarity of 94.2% with Australian serotype 1 and a minimum similarity of 67.4% with Australian serotype 15. However, at deduced amino acid level, it had maximum similarity of 99.7% and a minimum of 82.5% with Chinese serotypes 1, 2 and 4 and Australian serotype 15, respectively. Deduced amino acid sequence analysis of putative receptor binding domain (121-249) revealed all the nine hydrophilic domains to be conserved across the serotypes. Functional motifs present in VP7 protein were also conserved in almost all the BTV serotypes including Indian serotype 23. Phylogenetic analysis based on VP7 gene sequence revealed Indian BTV serotype 23 segregating into a monophyletic group along with Australian serotype 1 and Chinese serotypes 1, 2 and 4, indicating its close evolutionary relationship with these Australian and Chinese serotypes.

Isolated massive thyroid metastasis in lung cancer.

Metastasis to the thyroid gland is rare despite its rich vascular supply. Among the pulmonary malignancies metastasising to the thyroid, adenocarcinomas are the commonest. The appearance of metastatic disease in lung carcinoma indicates a poor prognosis and the average survival is two months. We report a 62-year-old woman with squamous cell carcinoma of lung metastatic to the thyroid that produced massive enlargement of the gland. The appearance of the secondary preceded the diagnosis of the primary malignancy by a few months. Ultimately, the patient succumbed to her disease.

The M and N genes-based simplex and multiplex PCRs are better than the F or H gene-based simplex PCR for Peste-des-petits-ruminants virus.

Nucleocapsid (N), matrix (M) and hemagglutinin (H) genes-based simplex PCRs and an N and M genes-based multiplex PCR were developed for detection of Peste-des-petits-ruminants virus (PPRV). The M gene PCR was the most sensitive, followed by N, H and an already described fusion (F) gene PCRs, as they could detect the virus in samples with titers of 101, 102, 104and 105 TCID50/ml, respectively. The multiplex PCR was as sensitive as the M gene PCR, but it had the advantage of differentiating PPRV from Rinderpest virus (RPV).

Melnick- needles osteodysplasty presenting with quadriparesis.

Melnick-Needles syndrome or osteodysplasty, a monogenic heritable bone dysplasia, is characterized by a typical facies and characteristic radiological findings. Less than 70 well-documented cases have been reported in literature; most of them were sporadic. We report the first case from Eastern India in an adolescent male, who had cranio-vertebral junction anomalies and presented with spastic quadriparesis at the age of 13 years.

Sequence analysis of the nucleoprotein gene of Asian lineage peste des petits ruminants vaccine virus.

The complete nucleotide sequence of the nucleocapsid (N) protein of the peste-des-petits ruminants vaccine virus (PPRV Sungri/96) belonging to the Asian lineage was determined. The gene was 1692 nucleotides in length and encoded a polypeptide of 525 amino acids. The PPRV Sungri/96 N gene has a nucleotide homology of 92% for PPRV Nigeria 75/1 to 55.5% for canine distemper virus. At amino acid level the homology was 94.1% with PPRV Nigeria 75/1, while with other morbilliviruses, PPRV Sungri/96 had only 71.4-64.9% amino acid identity. The phosphorylation prediction reveals eight conserved sites across morbilliviruses, whereas in the C-terminal portion of the protein the sites are not conserved. Phylogenetic analysis of different N proteins of morbilliviruses revealed five well-defined clusters as observed previously. To the best of our knowledge this is the first report describing the nucleocapsid gene sequence of PPRV Indian isolate.