A novel role of CD73-IFNγ signalling axis in human mesenchymal stromal cell mediated inflammatory macrophage suppression.

Introduction: Immunomodulation is the predominant mechanism via which Mesenchymal stromal cells (MSCs) mediate their therapeutic benefits. However, inconsistent success in numerous clinical trials warrants a better understating of the molecular mechanisms regulating their immunomodulatory properties. CD73, an ecto-5'-nucleotidase is abundantly expressed by MSCs, however its precise role in regulating their immunomodulatory properties is still elusive. The present study explored the role of CD73 in Interferon-gamma (IFNγ) sensing and in turn their ability to suppress "inflammatory" M1 macrophages. Materials and methods: CD73 knockdown MSCs (CD73-KDN) were initially assessed for expression of immunoregulatory molecules and IFNγ sensing ability by analysing expression of IFNγ signalling downstream targets such as pSTAT-1, Interferon-Stimulated Genes (ISG) and Indoleamine 2,3-dioxygnease (IDO), a prototypic IFNγ-induced immunomodulator. Next CD73-KDN MSCs were co-cultured with inflammatory M1 macrophages and evaluated for their ability to suppress them. To delineate the contributory role of CD73 and IFNγ signalling downstream target IDO, they were overexpressed independently in CD73-KDN MSCs and re-evaluated for their ability to suppress M1 macrophages. Results: CD73-KDN MSCs exhibited reduced expression of immunoregulatory molecules and were refractory to IFNγ signalling as indicated by attenuated expression of pSTAT-1, Interferon-Stimulated Genes (ISG) and Indoleamine 2,3-dioxygnease (IDO) upon IFNγ exposure. Since sensing of inflammation is critical for MSC mediated immunomodulation, CD73-KDN MSCs were functionally evaluated for their ability to immune-modulate "inflammatory" M1 macrophages wherein they failed to suppress M1 macrophages. Interestingly, ectopic expression of either CD73 or IFNγ signalling target IDO1 in CD73-KDN MSCs restored their ability to suppress M1 macrophages, establishing the importance of CD73-IFNγ signalling axis in MSC-mediated inflammatory macrophage suppression. Conclusion: The present study uncovers the unexplored role of CD73-IFNγ axis in MSC-mediated M1 macrophage suppression. MSC-educated macrophages are the actual immune-modulators at MSC transplant sites, thus CD73 can serve as a key immune-potency marker for benchmarking therapeutically relevant MSCs.

Casein (CSN) gene variants and parity affect the milk protein traits in crossbred (Bos taurus x Bos indicus) cows in sub-tropical climate.

Genotypes at four casein (CSN) loci-A26181G of CSN1S1, C6227T of CSN1S2, A8101C of CSN2, and A13104C of CSN3-along with non-genetic factors were studied for their effects on various milk protein traits in 100 crossbred cows with major inheritance of Holstein Frisian (Bos taurus) and Tharparkar (Bos indicus). Results demonstrated the presence of all CSN genotypes with a predominance of heterozygotes. At CSN2 (A8101C; His67Pro) locus, the A2 allele, desirable for human health, was present in 62% as heterozygous and 29% in homozygous condition. Among non-genetic factors, parity of the cows had a significant influence on the milk protein traits in these crossbreds. The genotypes at CSN1S1, CSN2, and CSN3 loci were found to influence (p<0.05 to 0.01) the casein and whey protein yields and composition traits. The casein index and total milk yield were most influenced by the CSN1S2 locus. The AA (A1 milk) genotype of CSN2 had significantly higher yields and percentages of casein and whey proteins. Positive influence of CC genotype of CSNS3 on milk proteins of was observed similar to Bos taurus cows; however, such influence of AA genotype of CSN2 locus may be distinctive to the crossbred cows maintained in subtropical condition. Overall, the results revealed the diverse effects of CSN genotypes on milk proteins in crossbred cattle.

Alternate PCR assays for screening of JH1 mutation associated with embryonic death in Jersey cattle.

Jersey haplotype (JH) 1, a stop-gain lethal mutation in the CWC15 gene, causes embryonic losses in Jersey cattle. Two PCR based assays using Amplification Refractory Mutation System (T-ARMS-PCR) and restriction fragment length polymorphism (PCR-RFLP) were developed for screening of the JH1 in cattle. During the screening, seven among 30 Indian Jersey bulls were identified as carriers of the mutant JH1 allele, the first time in the country. These PCR assays are economical, rapid and accurate; and can be used separately or in combination for screening and cross-validation of the JH1 carriers in Jersey cattle.

Development of PCR based assays for detection of lethal Holstein haplotype 1, 3 and 4 in Holstein Friesian cattle.

Holstein haplotype (HH) 1, 3 and 4 are lethal mutations, responsible for early embryonic losses in Holstein Friesian (HF) cattle, worldwide. Three PCR based assays - tetra Amplification Refractory Mutation System PCR, PCR primer induced restriction analysis and PCR-restriction fragment length polymorphism techniques for screening of HH1, 3 and 4, respectively were developed and validated. During screening, six among 60 HF bulls were found as carrier for either of three mutations. These PCR assays are highly accurate and reproducible and can be used for screening of the haplotypes in HF cattle.

In-vitro antioxidant and anti-hyperlipidemic activities of Blumea eriantha DC.

Alcoholic and water extracts of the stem and root of Blumea eriantha DC were prepared and evaluated for in-vitro antioxidant activity by methods like total reducing power, scavenging of us free radicals like as 1,2-diphenyl-2-picrylhydrazyl (DPPH), super oxide, nitric oxide, and hydrogen peroxide. The percentage scavenging effect of free radicals was compared with standard antioxidants like ascorbic acid and Butylated- hydroxyl anisole (BHA). Different extracts were also tested for anti-hyperlipidemic activity in triton WR-1339 (iso-octyl polyoxyethylene phenol)-induced hyperlipidemia in albino rats by determination of serum triglyceride like VLDL, LDL, HDL levels. Significant antioxidant activity was estimated in different methods, (p<0.01) for reducing power and (p<0.001) for scavenging DPPH, super oxide, nitric oxide, and hydrogen peroxide radicals. The different extracts having significant reduction (p<0.01) in cholesterol at 6 and 24 h and (p<0.05) at 48 h. There was significant reduction (p<0.01) in triglyceride level at 6, 24 and 48 h. There was significant increase (p<0.01) in HDL at 6, 24 and 48 h. From the VLDL was also significantly (p<0.05) reduced from 24 h and maximum reduction (p<0.01) results, it is clear that alcoholic and water extracts of Blumea eriantha DC can remarkably decrease plasma cholesterol, triglyceride, LDL, and VLDL and increase plasma HDL levels. In addition, the alcoholic and aqueous extracts have shown significant antioxidant activity.

POMA analyses as new efficient bioinformatics' platform to predict and optimise bioactivity of synthesized 3a,4-dihydro-3H-indeno[1,2-c]pyrazole-2-carboxamide/carbothioamide analogues.

A series of 43, 3a,4-dihydro-3H-indeno[1,2-c]pyrazole-2-carboxamide/carbothioamide analogues (D01-D43) were analysed using Petra, Osiris, Molinspiration and ALOGPS (POMA) to identify pharmacophore, toxicity prediction, lipophilicity and bioactivity. All the compounds were evaluated for anti-HIV activity. 3-(4-Chlorophenyl)-N-(4-fluorophenyl)-6,7-dimethoxy-3a,4-dihydro-3H-indeno[1,2-c]pyrazole-2-carboxamide (D07) was found to be the most active with IC(50)>4.83 µM and CC(50) 4.83 µM. 3-(4-Fluorophenyl)-6,7-dimethoxy-3a,4-dihydro-3H-indeno[1,2-c]pyrazole-2-carbothioamide (D41) was found to be the most active compound against bacterial strains with MIC of 4 µg/ml, comparable to the standard drug ciprofloxacin while 3-(4-methoxyphenyl)-6,7-dimethoxy-3a,4-dihydro-3H-indeno[1,2-c]pyrazole-2-carboxamide (D38) was found to be the most active compound against fungal strains with MIC 2-4 µg/ml, however less active than standard fluconazole. Toxicities prediction by Osiris were well supported and experimentally verified with exception of some compounds. In anticonvulsant screening, 3-(4-fluorophenyl)-N-(4-chlorophenyl)-6,7-dimethoxy-3a,4-dihydro-3H-indeno[1,2-c]pyrazole-2-carboxamide (D09) showed maximum activity showing 100% (4/4, 0.25-0.5h) and 75% (3/4, 1.0 h) protection against minimal clonic seizure test without any toxicity.

Structure-based design, synthesis and molecular modeling studies of thiazolyl urea derivatives as novel anti-parkinsonian agents.

Synthesis of 1-(substituted aryl)-3-(thiazol-2-yl)urea derivatives was undertaken as our efforts to discover novel antiparkinsonian agents with improved pharmacological profile in haloperidol-induced catalepsy and oxidative stress in mice. Furfuryl, 2- and/or 3-methoxy substituted phenyl derivatives emerged as potent agents. With exception of 2-chloro,5-trifluoromethyl substituted analog, halogen substituted derivatives exhibited moderate antiparkinsonian activity. The results of biochemical investigations from brain homogenate of mice outline the importance of neuroprotective/antioxidant therapy for Parkinson's disease (PD), supporting the notion that the oxidative stress may play a significant role in the pathophysiological mechanisms underlying PD. Molecular docking studies of these compounds with adenosine A(2A) receptor exhibited very good binding interactions and warrants further studies to confirm their binding with human A(2A) receptor for the design and development of potent antagonists. Parameters for Lipinski's rule of 5 were calculated computationally because pharmacokinetic and metabolic behaviors in the body often are linked to the physical properties of a compound. None of the synthesized compounds violated Lipinski's rule, making them suitable drug candidate for the treatment of PD.

Determination of the antiulcer properties of sodium cromoglycate in pylorus-ligated albino rats.

OBJECTIVES: To study the ulcer protective property of sodium cromoglycate in pylorusligated rats and the biochemical role in ulcer protection by various biochemical tests. MATERIALS AND METHODS: The ulcer protective effect of sodium cromoglycate was studied using a Pyloric Ligation Model using Wistar albino rats. The antiulcer effect of sodium cromoglycate 40 mg/kg b.w., i.p., was compared with the reference drug ranitidine 27 mg/kg b.w., i.p. The ulcer index was calculated and other biochemical parameters like free acidity, total acidity, pH, mucin, pepsin and volume of gastric juice were determined. RESULTS: Pylorus ligation showed a significant (P < 0.01) reduction in gastric volume, free acidity, total acidity and ulcer index as compared to the control. CONCLUSION: Sodium cromoglycate has activity equipotent to ranitidine.

The role of calcium, magnesium, and zinc in pre-eclampsia.

Pre-eclampsia is the most common medical complication of pregnancy associated with increased maternal and infant mortality and morbidity. Its exact etiology is not known, although several evidences indicate that various elements might play an important role in pre-eclampsia. This study was carried out to analyze and to compare the concentration of calcium, magnesium, and zinc in the serum of women with pre-eclampsia and in normal pregnant women. Fifty clinically diagnosed patients with pre-eclampsia (25 with mild and 25 with severe pre-eclampsia) and 50 normal pregnant controls were enrolled in this study. The serum calcium, magnesium, and zinc levels were estimated with an atomic absorption spectrophotometer. The mean serum levels of calcium, magnesium, and zinc in normal pregnant group were 2.45 +/- 0.18 mmol/L, 0.79 +/- 0.13 mmol/L, and 15.64 +/- 2.4 micromol/L, respectively, while in mild pre-eclamptic group, these were 2.12 +/- 0.15 mmol/L, 0.67 +/- 0.14 mmol/L, and 12.72 +/- 1.7 micromol/L, respectively. Serum levels in severe pre-eclamptic group were 1.94 +/- 0.09 mmol/L, 0.62 +/- 0.11 mmol/L, and 12.04 +/- 1.4 micromol/L, respectively. These results indicate that reduction in serum levels of calcium, magnesium, and zinc during pregnancy might be possible contributors in etiology of pre-eclampsia, and supplementation of these elements to diet may be of value to prevent pre-eclampsia.

Syntheses and anti-inflammatory activity of diphenylamine-2,2'-dicarboxylic acid and its metal complexes.

Diphenylamine-2,2'-dicarboxylic acid and its Cu(II), Ni(II), Co(II) and Zn(II) complexes have been synthesized and characterized by their elemental analyses, molecular weight determination, molar conductance, infrared and electronic spectra and magnetic measurements. The Zinc complex was tested by different methods for its anti-inflammatory activity and found to be equipotent to naproxen and ibuprofen, though at higher doses.

An ongoing Confidential Enquiry into asthma deaths in the Eastern Region of the UK, 2001-2003.

INTRODUCTION: The Eastern Region Confidential Enquiry into asthma deaths started in 2001. It incorporates the Norwich and East Anglian Enquiries started in 1988 and 1992, respectively. The aim of this study was to analyse all asthma deaths in the Eastern region between 2001 and 2003, to elicit any factors contributing to the patients' deaths, and to make comparisons with the previous Norwich and East Anglian data. METHOD: Patient details were obtained for all deaths in the Eastern Region under the age of 65 with asthma recorded in the first part of the death certificate. Patients' notes were reviewed by members of the Working Group - a consultant chest physician and a general practitioner (GP). In most cases, the patient's GP was contacted. Data were obtained on the patients' asthma care, asthma severity, terminal attack, psychosocial and behavioural factors, allergies, precipitating factors, and post-mortem findings. The quality of medical care was assessed and compared with national guidelines. RESULTS: Total study population was 5.25 million. Only 57/95 notified deaths (60%) were confirmed as asthma deaths. 311 asthma deaths have been studied between 1988 and 2003. In 2001-2003, male:female ratio was 3:2. Further data were unavailable on three cases. 53% of patients had severe asthma and 21% moderately severe disease. In 19 cases (33%) at least one significant co-morbid disease was present. Monthly death rates peaked in August, with a smaller peak in April. In 11 cases (20%), mostly males aged under 20, the final attack was sudden and 10/11 occurred between April and August. In 81% of cases there were significant behavioural and/or psychosocial factors such as poor compliance (61%), smoking (46%), denial (37%), depression (20%) and alcohol abuse (20%). The overall medical care of the patient was appropriate in 33% of cases. CONCLUSIONS: Between 1988 and 2003 there was a downward trend in asthma mortality rate in East Anglia. In 2001-2003, misclassification of deaths attributed to asthma was still common. Most patients who die of asthma have severe asthma. In 81% of cases, behavioural and psychosocial factors contributed to the patient's death. In 80% of deaths the final attack was not sudden, and may have been preventable. Almost all sudden deaths occurred between April and August, suggesting a seasonal allergic cause. In two-thirds of asthma deaths, medical management failed to comply with national guidelines. 'At-risk' asthma registers in primary care may improve recognition and management of 'at-risk' patients.

Studies on the Antifungal Activity of Pterocarpus Marsupium: a Clinical Evaluation.

Pmarsupium (Hindi Bijasar) is a powerful astringent and is used chiefly in diarrhoeas. It is also an useful remedy for diabetes mellitus and various skin diseases as mentioned in literature. In a blind clinical trial, the usefulness of this drug as a topical agent against T.cruris and T. corporis was elevated. The drug yielded good response within 3 days of the first application.