Effect of asiatic acid supplementation in tris-extender on post-thaw functional competence, antioxidant enzyme activity and in vivo fertility of bull sperm.

Reactive oxygen species at supra-physiological levels trigger oxidative stress during cryopreservation, which can be neutralised by incorporating suitable antioxidants into the semen extender medium. This study was intended to explore the effect of asiatic acid (AA) as an antioxidant in semen extender on frozen-thawed sperm quality and in vivo fertility of bull sperm. Semen was collected from Holstein Friesian bulls for 10 consecutive weeks (total ejaculates = 60). Semen was cryopreserved with a Tris citric acid egg yolk-based extender supplemented with 0 (control), 20, 40, 60, and 100 µM AA. The supplementation of the extender with 40 and 60 µM AA improved (p < 0.05) post-thaw motility kinematics, plasma membrane integrity, acrosome integrity, sperm viability, and DNA integrity of bull sperm. Mitochondrial membrane potential was high (p < 0.05) with 60 µM of AA concentration in extender media. The catalase activity in seminal plasma was maintained (p < 0.05) when semen was added with 20, 40, and 60 µM of AA. The in vivo fertility was found to be significantly high with the semen extended with 60 µM AA. Conclusively, this study showed that AA supplementation in semen extender significantly improved sperm motility kinematics and cell integrity, conserved antioxidant enzyme activity, and improved in vivo fertility.

In vivo analysis the effect of antibiotic growth promoters (AGPs), Oxytetracycline di-hydrate and Tylosin phosphate on the intestinal microflora in broiler chicken.

The study was aimed to analyse the effects of antibiotic growth promoters (AGPs), Oxytetracycline di-hydrate and Tylosin phosphate on the intestinal microflora in broiler chicken. The AGPs were provided in different concentrations solely or in combinations for 42 days of rearing. Faecal samples were collected from the intestine (duodenum, jejunum and caeca) of broiler chicken on 14th, 28th and 42nd days of trial. Samples were cultured on different selective medium and bacterial identification was performed by different biochemical and molecular diagnostic tools. Results showed a significant effect of AGPs on the growth of pathogenic microorganisms such as Escherichia coli and Clostridium perfringens in the intestine. Interestingly, an impaired growth was observed for both bacterium showing a significant effect (P<0.05) of AGPs on E. coli and C. perfringens on day 14th, 28th, and 42nd. This effect was observed solely and in combination while using AGPs. Data further showed that the effect was more prominent in combination and with an increase concentration of AGPs. Remarkably, no impairment was seen on the growth of L. reuteri at different sites of intestine and duration (14th, 28th, and 42nd days). The results showed that the use of AGPs in diet has no harmful effect on beneficial bacteria, however, an impaired growth was seen on the harmful bacteria. It is suggested that a combination of AGPs (OXY-1.0+TP-0.5) is economical and have no harmful effect on the broiler chicken. The use of AGPs in a recommended dose and for a specific period of time are safe to use in poultry both as growth promoter and for the prevention of diseases.

Epidemiology of Van der Woude syndrome from mutational analyses in affected patients from Pakistan.

Mutations in IRF6 cause Van der Woude syndrome (VWS), one of the most common syndromes associated with cleft lip (CL) with or without cleft palate (CP). The presence of pits on the lower lip of patients is the most characteristic feature of the syndrome. We have identified three novel and seven previously reported IRF6 mutations in 12 of 16 unrelated families segregating VWS from Pakistan. The three newly identified mutations include a frameshift (c.568delG) and two missense mutations c.295G>A (p.G99S) and c.1219T>C (p.S407P). Recent functional studies on IRF6 and the three-dimensional structure of IRF5 carboxy (C) terminus, a protein encoded by a paralog of IRF6, shed light on the p.S407P substitution. Additionally, the identification of the same mutations responsible for VWS in Pakistan, as reported in other global populations worldwide, marks these residues as mutational hotspots and indicates their essential role in structural stability or function of IRF6. This is the first study of VWS in Pakistan and we estimate that 1 in 100 patients with CL with or without CP (CL/P) are affected in the Pakistani population predominantly from the Punjab area.

Wegener's granulomatosis presenting as diabetes insipidus.

Pituitary gland involvement in Wegener's granulomatosis (WG) occurs most commonly in the form of central diabetes insipidus (CDI). However, CDI as a presenting manifestation of WG is very rare. We report two such cases; one of them had multi-organ involvement at presentation, while other developed it during follow-up. CDI was reversible following cytotoxic drug therapy in one of them.

The effect of aspirin and two nitric oxide donors on the infarcted heart in situ.

Nitric oxide (NO) donors are heterogeneous substances which release NO, a biologically active compound. NO released by nitric oxide donors has important effects on the circulation by causing vasodilation, diminishing myocardial contractile force, inhibiting platelet aggregation, and counteracting the effects of thromboxane A2. In the infarcted heart, activation of the inducible form of nitric oxide synthase (iNOS) and the formation of prostacyclin and thromboxane A2 by cyclooxygenase (COX) were increased. Myocardial infarction also resulted in increased myocardial NO production. Aspirin (acetylsalicylic acid. ASA) at low concentration (35 mg/kg/day) fails to change iNOS production, in contrast to higher dose (150 mg/kg/day) which, as previously shown, inhibits iNOS activity. ASA at all doses also suppresses myocardial prostanoid formation because of inhibition of COX. Recently, two NO donors have been synthesized: NCX 4016 and Diethylenetriamine/NO (DETA/NO). NCX 4016 combines an NO-releasing moiety with a carboxylic residue via an esteric bond. We describe here that NCX 4016 (65 mg/kg/day) increased prostacyclin and thromboxane A2 production in the infarcted heart muscle, overcoming the inhibitory effects of ASA. As a result of nitric oxide release, oxidation products of NO (NO2- and NO3-; NOx) in arterial blood rose following administration of NCX 4016. On oral administration, NCX 4016 did not change systemic arterial pressure. The effects of a single NO donor, DETA/NO (1.0 mg/kg/day) on the infarcted heart were also investigated On intravenous administration, the compound increased NO concentration in arterial blood slightly but to a lesser degree than NCX 4016. Like NCX 4016, it raised myocardial production of prostacyclin and thromboxane A2 in the infarcted heart. However, it caused a severe fall in blood pressure. These findings demonstrate that newly-synthesized NO donors release nitric oxide in situ and increase myocardial production of prostanoids. NCX 4016 has therapeutic potential because it can be orally administered, lacks hypotensive effects, increases blood levels of nitric oxide and myocardial prostacyclin production.

Erythema nodosum leprosum in histoid leprosy: a case report.

A 28-year-old married female developed histoid papules and nodules de novo over her face, extremities, back, buttocks and thighs. She had developed erythema nodosum leprosum lesions without any antileprosy treatment. Histopathology from a histoid nodule showed well defined nodular collections of plump, spindle-shaped histiocytes. A few globi were also seen with Ziehl Neelson staining. Leucocytoclastic vasculitis was present in the ENL lesion. The CD4:CD8 ratio was 1.5:1.

Clinico-bacteriological study of pyodermas in children.

One hundred cases of pyodermas in children were investigated clinically and bacteriologically. Nasal and throat swabs from all cases were subjected to bacteriological examination. Most of the children (42%) were in the 1-4 year age group. The majority (58%) belonged to lower socio-economic groups with poor standards of hygiene. A history of over-crowding was obtained from 87% of cases, 82% were undernourished. Most of the children (68%) reported during the hot and humid months of June, July, August, and September. Primary pyodermas were observed in 72% of the children, and secondary pyodermas in 28%. Impetigo was the commonest primary pyoderma (48.61%); among secondary pyodermas, infected scabies was noted predominantly (42.86%). The face and legs were more commonly involved. Bacteriological cultures from pyoderma lesions revealed a single microorganism in the majority of the patients (84%). Staphylococcus aureus was isolated in pure culture from 48% and pure beta-hemolytic streptococci from 36%. A combination of both was obtained from 16%. No other organism was isolated from any case. A similar pattern was also observed in cultures from the nose and throat. Only 46 out of the 64 strains (84.3%) of Staphylococcus aureus isolated from pyoderma were typable. The majority (39.1%) showed a mixed pattern of phages; the second commonest was the non-allocated phage type (30.4%). Nasal flora had more of the non-allocated phage type (50%); two out of the three strains (66.6%) isolated from the throat showed a mixed pattern. All the strains of beta hemolytic streptococci, isolated either from lesions of pyoderma, nose, or throat belonged to group A. Staphylococcus aureus and showed a high sensitivity to netilmycin (100%), ofloxacin (98.4%), amoxycillin/clavulanic acid (96.9%), ciprofloxacin (89.1%) and gentamycin (84.4%) but a high resistance to penicillin (85.9%). A greater correlation was noted between nasal flora and organisms causing pyodermas. A change in the pattern of organisms causing pyodermas in children and their antibiotic sensitivities in this part of the globe has been observed in this study. The role of endogenous nasal and throat flora in the causation of pyodermas has also been highlighted.

Familial reactive perforating collagenosis: a case report.

Reactive perforating collagenosis is characterised by trans-epidermal elimination of collagen and is hypothesized to be both autosomally dominant and recessive. We report a family in which two brothers and a sister had lesions of reactive perforating collagenosis.

Three unusual siblings with Harlequin icthyosis in an Indian family.

Harlequin fetuses occurring as three siblings in an Indian family are described here. All three were preterm, low birth weight, and did not survive. There was no history of consanguinous marriage in the parents or in the family. Thus autosomal recessive inheritance appears to be a remote possibility, although not impossible or, as recently described, these recurrent harlequin fetuses could be the result of new dominant mutations with parental mosaicism.

Effect of cyclooxygenase-2 inhibitor (celecoxib) on the infarcted heart in situ.

Several attempts have been made to replace aspirin with compounds without gastric toxicity; a cyclooxygenase-2 (COX-2) inhibitor, celecoxib, and a nitric oxide-aspirin, NCX-4016, have been developed for this purpose. This paper compares effects of celecoxib, NCX-4016 and aspirin on production of prostacyclin (PGI2) and thromboxane A2 (TXA2) and activation of the inducible form of nitric oxide synthase (iNOS) in infarcted heart in situ. Aspirin was most effective in reducing myocardial PGI2 synthesis and formation of TXA2. Myocardial effects of celecoxib resemble those of NCX-4016, although the two compounds have different modes of action.

Production of prostanoids and nitric oxide by infarcted heart in situ and the effect of aspirin.

The production of prostacyclin (PGI2) and thromboxane A2 (TXA2) in infarcted and noninfarcted portions of the rabbit heart was studied prior to and following administration of acetylsalicylic acid (aspirin). Aspirin was administered intravenously (iv) as water-soluble Aspisol, d-lysinmono (acetylsalicylate) (Bayer, Leverkusen, Germany) into an ear vein. A branch of the left circumflex coronary artery was ligated. The animals were divided into three groups. The first group received 150 mg/kg/day of aspirin (75 mg/kg of aspirin every 12 h, n = 10). The first administration of aspirin was 1 h after ligation of the coronary artery and the last injection was 1 h before euthanasia. The second group received 5 mg/kg/day of aspirin (every 24 h, n = 10). A separate group of rabbits not receiving aspirin served as controls (n = 12). Two days following onset of ischemia, inducible form of nitric oxide synthase (iNOS) was measured in heart muscle and the oxidation products of nitric oxide (nitrite, NO-2 plus nitrate, NO-3: their sum referred to as NOx) were determined in arterial and coronary venous blood. Concentrations of both PGI2 and TXA2 were elevated in the infarcted portions of the heart compared to the noninfarcted regions. Formation of prostanoids was accompanied by increased activation of iNOS. Both doses of aspirin diminished the concentrations of PGI2 and TXA2 in infarcted heart muscle; in contrast, small doses of aspirin failed to influence myocardial iNOS activity. Apparently small doses of aspirin changed the relationship of iNOS to cyclooxygenase (COX). Coronary arterial-venous difference of NOx and myocardial iNOS activity showed parallel increases. Diminution of prostacyclin by aspirin can damage gastric mucosa and interfere with vasodilatation. Since NO counters these deficiencies, a combination of aspirin with a nitric oxide donor may be advantageous.