Naloxone blocks the beneficial effects of aqueous extract of Murraya koenigii (L.) Spreng leaves in models of pain.

AIM: This study investigated the antinociceptive effects of aqueous extract of Murraya koenigii (AEMK) leaves (200, 400 and 800 mg/kg, orally) on animal models of acute and persistent pain and its modulation by naloxone. MATERIALS AND METHODS: Antinociceptive effects were assessed using tail-flick, hot plate and formalin tests in mice. To differentiate between central and peripheral antinociceptive effect of AEMK, naloxone (2 mg/kg) was administered along with the 800 mg/kg dose of extract. Morphine was used as a standard drug. RESULTS: AEMK and morphine significantly increased the tail-flick latency (tfl) and paw licking/jumping latency in tail-flick and hot plate tests, respectively, in comparison to control. Also, in both the tests AEMK and morphine significantly increased the AUC0-120 min. In formalin test, AEMK (400 mg/kg and 800 mg/kg) and morphine significantly reduced licking time in both early and late phases in comparison to control. CONCLUSIONS: Thus, in all three pain models AEMK showed antinociceptive effect, which was blocked by naloxone suggesting the involvement of opioidergic central mechanism.

N-Methyl-D-aspartate receptor modulators block hyperalgesia induced by acute low-dose morphine.

1. Following opioid-induced antinociception in mice, hyperalgesic responses may be observed. The present study was designed to evaluate the effect of different N-methyl-d-aspartate (NMDA) receptor modulators (magnesium, dextromethorphan, d-serine) on the development of morphine-induced hyperalgesia in mice. The tail-flick test was used to assess the effects of morphine alone and in combination with the NMDA receptor modulators. 2. Administration of a single low dose (2 mg/kg) of morphine to mice produced antinociception that was followed by hyperalgesia. 3. Administration of magnesium sulphate (5 mg/kg) and d-serine (10 mg/kg) alone produced a transient antinociceptive response, whereas dextromethorphan (10 mg/kg) alone produced a prolonged antinociceptive response that had a relatively delayed onset after 4 h. 4. When coadministered with morphine, the NMDA receptor blockers magnesium (2 mg/kg) and dextromethorphan (2 and 5 mg/kg) and the NMDA receptor agonist d-serine (2, 5 and 10 mg/kg), maintained the duration of the antinociceptive response to morphine and inhibited the development of the hyperalgesic response. Coadministration of dextromethorphan (10 mg/kg) with morphine produced antinociception at 30-120 min and at 4-24 h. 5. The results of the present study suggest that coadministration of low-dose NMDA receptor antagonists, as well as the NMDA receptor agonist d-serine, with morphine can inhibit morphine-induced hyperalgesia.

Cell adherence--a determinant of virulence in Yersinia enterocolitica.

The adherence of pathogenic bacteria to eukaryotic cells plays a central role in their ability to colonize the mucosal epithelial surfaces. The adherence by Y. enterocolitica to the mucosal surface of the gut is the initiating event leading to penetration of mucosa. Adhesion of 10 probable pathogenic and one non-pathogenic isolate was studied using ileum and colon epithelial cells of mouse for 90 minutes. Adhesion study revealed that isolates of Y. enterocolitica had a good adhesive property while non pathogenic showed negligible adherence. All isolates showed better adherence to colon epithelial cells. The organism continued to be excreted in faeces up to 8-10 days after oral feeding. Adhesion positive isolates were found to be virulent when tested in mice for diarrhoea and death. Adhesion was found to be thermoregulated.

Juvenile hyaline fibromatosis in siblings.

Juvenile hyaline fibromatosis (JHF) is a rare, autosomal recessively inherited disorder. We report two siblings with multiple large tumors on the scalp, translucent papules on the nape of the neck, hypertrophic gingiva, and severe flexural contractures of large joints. The histopathology from the skin lesions showed features characteristic of juvenile hyaline fibromatosis. The cases are being reported on account of the extreme rarity of the condition.

Florid cutaneous papillomatosis with adenocarcinoma of stomach in a 35 year old male.

Various paraneoplastic dermatoses may be seen in association with underlying visceral, especially gastrointestinal, malignancy. Florid cutaneous papillomatosis describes the sudden appearance of multiple acuminate keratotic papules that morphologically resemble viral warts. It may be seen in association with acanthosis nigricans and/or the sign of Leser Triotalat. We report a 35-year-old male with extensive seborrhoeic keratoses and florid cutaneous papillomatosis. Unusually marked verrucous changes caused disfigurement of the hands and feet. The patient also reported dyspepsia, abdominal distention and weight loss 6 months prior to the development of cutaneous lesions. Gastroscopy revealed a large growth in the stomach. Histopathology of the tumor showed features of adenocarcinoma.

Occupational contact dermatitis among the traditional 'tie and dye' cottage industry in Western Rajasthan.

BACKGROUND: Dyeing is an age-old process and forms an integral part of textile industries. Tying is a process by which a particular part of cloth is prevented from the process of dyeing. The skin diseases in workers engaged in the 'tie and dye' industry have not been extensively studied. AIMS: To study the prevalence of contact dermatitis among workers engaged in the 'tie and dye' industries in and around Jodhpur (Western Rajasthan). METHODS: One thousand three hundred workers engaged in 'tie and dye' work were evaluated for occupation-related dermatitis. Those with skin lesions were subjected to patch tests using 2% aqueous solution of the dyes and chemicals commonly used by them. These included direct dyes, VAT dyes, sulfur dyes and azo dyes. Fifty workers without skin lesions served as controls. RESULTS: One hundred patients (7.69%) had dermatitis involving the exposed sites, mainly the hands and forearms. Eighty-one patients showed positive reactions to one or more dyes, most commonly Red RC base (azo dye), followed by naphthol. CONCLUSION: Red RC base and naphthol were the commonest allergens in the 'tie and dye' industry.

Comparison of efficacy, safety and brain derived neurotrophic factor (BDNF) levels in patients of major depressive disorder, treated with fluoxetine and desvenlafaxine.

This randomized, open label, prospective, observational study compared clinical efficacy, safety alongwith plasma BDNF levels in outpatients of depression treated with fluoxetine and desvenlafaxine. Patients (aged 18-60 years) with moderate to severe major depressive disorder (MDD) diagnosed by DSM-IV criteria, and Hamilton Rating Scale for Depression (HAM-D) score ≥14, who were prescribed fluoxetine or desvenlafaxine were included (n=30 in each group). Patients were followed up for 12 weeks for evaluation of clinical efficacy, safety along with BDNF levels. In the fluoxetine group, HAM-D scores at the start of treatment was 19±4.09 which significantly (p<0.05) reduced to 9.24±3.98 at 12 weeks. In the desvenlafaxine group, HAM-D scores at the start of treatment was 18±3.75 which significantly (p<0.05) reduced to 10±3.75 at 12 weeks. The BDNF levels in the fluoxetine group were 775.32±30.38pg/ml at the start of treatment which significantly (p<0.05) increased to 850.3±24.92pg/ml at 12 weeks. The BDNF levels in the desvenlafaxine group were 760.5±28.53pg/ml at the start of treatment which significantly (p<0.05) increased to 845.8±32.82pg/ml at 12 weeks. Both the antidepressants were found to be safe and well tolerated. The efficacy and the safety profile of desvenlafaxine is comparable to fluoxetine in patients of MDD. BDNF levels were significantly increased post-treatment with both the antidepressive agents. Whether BDNF may have a prognostic value in predicting treatment response to antidepressant drugs needs to be investigated in a larger patient population.

Purification and properties of levanase from Rhodotorula sp.

Levanase, a slime dissolving enzyme of Rhodotorula sp., was purified to approx. 26-fold by ammonium sulphate precipitation, DEAE and gel filtration (Sephacryl S-200) chromatography. The molecular mass of the enzyme was 39 kDa. The purified levanase showed maximum activity at pH 6.0 and 40 degrees C. Enzyme was quite stable at 4 degrees C and at pH 5.5 to 6.5. Hg2+ at a level of 10 mM completely inhibited the levanase activity, while 2-mercaptoethanol at the same concentration showed a 2.93-times increase in activity. In addition to levan, the enzyme also showed substrate specificity towards inulin.

Cytotoxic T-cell, delayed type hypersensitive and listeriolysin O responses in experimental bovine listeriosis.

Five different antigens of Listeria monocytogenes NCTC 11994 were assayed for their lymphoproliferative, cytotoxic, humoral and delayed type hypersensitivity (DTH) responses in experimentally infected calves. The antigens used were culture filtrate antigen (CFA), listeriolysin O (LLO), sonicated antigen (SA), live antigen (LA) and heat killed antigen (HKA). CFA was most lymphoproliferative when assayed in vitro. In an autologous monocyte cytotoxicity assay, soluble protein as well as bacterial cells elicited cytolytic responses, however, LA and LLO primed monocytes showed a higher cytotoxic effect. The expression of cell surface markers of lymphocyte subpopulations was almost identical in experimentally infected as well as non-infected calves. SA elicited the highest humoral and DTH responses. LLO being a major virulence factor with appreciable humoral as well as cellular responses may be used as a candidate antigen to develop a reliable diagnostic immunoassay.

Reactive nitrogen intermediates production from naive and activated monocytes by extracts of Mycobacterium bovis BCG.

Extracts of Mycobacterium bovis strain BCG were assessed for in vitro activation of monocytes to produce reactive nitrogen intermediates. The culture filtrate of M. bovis BCG was a strong inducer of nitrite production while live BCG and sonicated antigens were also potent inducers. Other extracts activated monocytes which showed an increase in nitrite production after in vitro BCG infection.

Induction of immune responses in cattle with a DNA vaccine encoding glycoprotein C of bovine herpesvirus-1.

A DNA vaccine expressing glycoprotein C (gC) of bovine herpesvirus-1 (BHV-1) was evaluated for inducing immunity in bovines. The plasmid encoding gC of BHV-1 was injected six times intramuscularly or intradermally into calves at monthly intervals. After immunization by both routes neutralizing antibody and lymphoproliferative responses developed. The responses in the intradermally immunized calves were better than those in calves immunized intramuscularly. However, the intradermal (i.d.) route was found to be less efficacious when protection against BHV-1 challenge was compared. Following intranasal BHV-1 challenge, all immunized calves demonstrated a rise in IgG antibody titre on day 3, indicating an anamnestic response. The control non-immunized calf developed a neutralizing antibody response on day 7 post-challenge. The immunized calves showed a slight rise in temperature and mild clinical symptoms after challenge. The intramuscularly immunized calves showed earlier clearance of challenge virus compared with intradermally immunized calves. These results indicate that DNA immunization with gC could induce neutralizing antibody and lymphoproliferative responses with BHV-1 responsive memory B cells in bovines. However, the immunity developed was not sufficient to protect calves completely from BHV-1 challenge.

Effect of in vitro monocyte activation by Listeria monocytogenes antigens on phagocytosis and production of reactive oxygen and nitrogen radicals in bovines.

Macrophages by virtue of their phagocytic and antibacterial activities play an important role in the host resistance to intracellular pathogens including Listeria monocytogenes. However, the precise killing mechanism adopted by macrophages in the case of L. monocytogenes and the role of macrophage activation in bacterial killing are still unclear. In the present investigation, different antigens of pathogenic L. monocytogenes and three non-specific activators, namely, Lipopoly-saccharide (LPS), Phorbol Myristate Acetate (PMA) and Concanavalin A (ConA) supernatant were studied to adjudge their efficacy with regard to in vitro activation of bovine monocytes in terms of the production of reactive nitrogen intermediates (RNI), reactive oxygen intermediates (ROI) and their phagocytic index (PI). Of all the five antigens of L. monocytogenes, namely, viable bacteria used as live antigen (LA), heat-killed antigen (HKA), sonicated antigen (SA), culture filtrate antigen (CFA) and listeriolysin-O (LLO), LA turned out to be the best activator of monocytes for RNI as well as ROI production. In the PI assay, of the three antigens, that is, CFA, SA and LLO, CFA was found to be the best activator of phagocytosis followed by LLO and SA. Among non-specific activators, PMA induced the highest H2O2 production whereas LPS caused the maximum increase in RNI and PI production. On activation by different antigens of L. monocytogenes, the peripheral blood mononuclear cells (PBMCs) from infected animals produced significantly higher RNI than those from non-infected animals indicating the involvement of immune T-cells.

Virulence of silver-resistant mutant of Klebsiella pneumoniae in burn wound model.

A silver-resistant mutant of Klebsiella pneumoniae B-5 was produced by passaging in nutrient broth containing graded concentrations of silver nitrate up to 150 ppm. The development of silver resistance in the strain resulted in rough colonies, decrease in cell size, carbohydrate content and change in klebocin pattern. The virulence of the AgR strain as checked by the burn wound model decreased as the mutant could not establish itself in the skin and spleen of the animals and the organism was cleared more efficiently by human lymphocytes than the parent AgS strain.

Induction of nitric oxide in bovine peripheral blood mononuclear cells by bovine herpesvirus 1.

Bovine peripheral blood mononuclear cells (PBMCs) and monocytes were stimulated with bovine herpesvirus 1 (BHV-1), LPS and concanavalin A (Con A) to produce L-arginine-dependent nitric oxide (NO) in vitro. NO was detected as early as 12 hrs and up to 72 hrs post stimulation (p.s.). The NO from lipopolysacharide (LPS)-stimulated PBMCs and monocytes was found to exhibit antiviral effect against BHV-1. The anti-BHV-1 effect was inhibited with N(omega)-methyl L-arginine indicating involvement of inducible NO synthase (iNOS) in NO production.

Identification of bovine sperm specific polypeptides reactive with antisperm antibodies.

The present study was taken to characterize molecular weights of sperm specific polypeptides antigenic to rabbits and calf with the aim to assess their immunoreactivity with IgG antibodies in sera from immuno-infertile cows. Seropositivity for antisperm IgG antibodies in 75 repeat breeder and 15 pregnant control cattle was tested by cellular ELISA using washed spermatozoa antigen from 4 bulls. Molecular weights of bovine sperm polypeptides antigenic to rabbit and calf were determined by 10% SDS-PAGE and Western blotting. Molecular weights of sperm peptides reactive with sera from immuno-infertile cows were also determined. Seropositivity of antisperm IgG antibodies for bull I, II, III and IV was 23.6, 14.6, 26.6 and 20%, respectively. A total of 16 polypeptides were discernible on gel. Out of these, 7 polypeptides were immunoreactive with sera from hyperimmunized rabbits as compared to 3 poly-peptides which reacted with sera from hyper-immunized calf. Only two polypeptides were reactive with sera from immuno-infertile cows. Variable number of sperm polypeptides and their immunoreactivity have been reported in different species. Antigenicity of different polypeptides in sperm needs further investigations.

Kinetics of interferon-gamma production and its comparison with anti-listeriolysin O detection in experimental bovine listeriosis.

The kinetics of the production of interferon gamma (IFN-gamma) in whole blood culture and its comparison with anti-listeriolysin O (ALLO) detection by ELISA were studied during oral infection of calves with Listeria monocytogenes. Culture filtrate antigen (CFA), listeriolysin O (LLO), and sonicated antigen (SA) were used to prime the peripheral blood mononuclear cells (PBMCs) and the plasma from orally infected calves. IFN-gamma and ALLO appeared as early as day 7 of an oral infection. IFN-gamma was detected earlier with LLO than with SA. The Max50 interleukin (IL-2) activity and IFN-gamma estimated in the culture supernatant from PBMCs primed in vitro with different antigens of L. monocytogenes revealed high induction of IL-2 and IFN-gamma by CFA, LLO and live antigen. IFN-gamma assay and ALLO detection were used for testing cases of repeat breeding in dairy cattle. It appeared that detection of IFN-gamma employing LLO can be used to diagnose listerial infections.

Seropositivity for intracellular bacterial infections among abattoir associated personnels.

Studies were carried out to detect the antibodies against zoonotic intracellular bacterial infections viz., brucellosis, listeriosis and tuberculosis in abattoir associated personnel employing dot-ELISA. Out of 165 serum samples tested 25.5, 40.0 and 10.9 per cent were detected as positive for brucellosis, listeriosis and tuberculosis, respectively. Immunodetection of these occupationally exposed persons for IgM as well as IgG antibodies revealed positivity for IgM, IgG and IgM + IgG in 8.5, 17.6 and 13.9 per cent for listeriosis and 5.5, 2.4 and 3.0 for tuberculosis respectively. Antibodies against brucellosis and listeriosis both were detected in 10.9 per cent persons, while 6.7 per cent persons were positive for both listeriosis and tuberculosis. No person was found positive with both brucellosis and tuberculosis. All the three infections were detected in 3.6 per cent persons.

Histopathological and ultrastructural alterations of induced Yersinia enterocolitica infection in mice.

Yersinia enterocolitica is an enteric bacterium and infections by this organism are mostly foodborne. It has been implicated to cause enterocolitis, terminal ilitis. diarrhoea, mesenteric lymphadenitis and arthritis in man. Due to paucity of information regarding histopathological and specially ultrastructural alterations in tissues affected, this study was planned with mice as the experimental model. Nine pathogenic Y.enterocoliticaisolates were used to infect 80 albino mice by oral and intraperitoneal route. Pathological alterations were studied by light and electron microscopy. Histopathological examination of intestines showed severe edema, purulent enteritis, goblet cell hyperplasia infiltration of mononuclear cells, thickening of mucosa and necrosis of the tips of villi. Liver showed congestion, hepatocellular degeneration and necrosis, atrophy of hepatocytes and microabcesses. The lungs revealed congestion, edema, haemorrhage and purulent ronchopneumonia, while kidneys showed mild necrotic changes and bacterial emboli in glomeruli. Ultrastructural changes were indicative of mitochondrial degeneration and their loss in kidneys, membranous degeneration with formation of myelin figures in lungs and disorganization, disruption and bleb formation of microvilli in intestines. Y.enterocolitica caused significant histopathological and ultrastructural alterations in experimentally infected mice. Variation in pathogenicity of different strains of Y.enterocolitica was also observed.

Familial syringoma.

Three patients with a family history of syringomas seen over a 12-month period are reported. Familial syringomas may not be as rare as generally believed.

Pityriasis lichenoides chronica in association with Tubercular Lymphadenitis.

A patient with pityriasis lichenoides chronica (PLC) during the course of her skin disease developed tuberculosis of cervical lymph nodes. Anti-tuberculosis treatment resulted in remission of PLC. Mycobacterium tuberculosis could be one of the triggering agent in pityriasis lichenoides.