Age-related changes in the phenotype of microglia

Microglia play a central role in immune surveillance and modulation of neuroinflammation as well as playing a role in neurodevelopment. Microglia involve in the development of pathological pain in adults but not early in life. However little detail is known about the changing phenotype of microglia during development. We examined age-related changes in microglia following activation with pathogen- and damage- associated molecular patterns (PAMPs/DAMPs). Microglial cultures were prepared from neonatal postnatal day (P1) and adult (P40) rat brains and spinal cords. Immunocytochemistry, qRT-PCR and functional assays were used to identify age-related differences. Adult microglia display a pro-inflammatory immune profile characterized by significantly increased IL-1β mRNA levels in response to PAMPs and DAMPs. In contrast, IL-1β mRNA in neonatal microglia showed a slight increase after stimulation with DAMPs. Anti-inflammatory gene expression was significantly increased in neonatal microglia relative to adult microglia. Compared to adult microglia, neonatal cells had increased phagocytic activity when unstimulated and following activation with LPS and ATP. Moreover, the nuclear receptor Nurr1 may play a major role in reducing pro-inflammatory signalling and promoting the anti-inflammatory phenotype in neonatal microglia. Nurr1 isoforms are differentially expressed in neonatal and adult microglia, with the Nurr1a isoform being significantly elevated in neonatal cells. Using lentiviral vector-mediated expression of Nurr1 isoforms, we also show that over-expression of TINUR, a splice variant of Nurr1, in neonatal and adult microglia attenuates inflammation by trans-repression the IL-1β expression and trans-activation the IL-10 gene expression following ATP exposure. Together, these data provide evidence for age-related difference in microglial function during postnatal development. In addition, these findings demonstrate insight into the mechanisms by which Nurr1 might act, and suggest potential therapeutic targets for the treatment of neuro-inflammatory diseases

Allelic frequency of PON1 Q192R, CYP2C19*2 and CYP2C19*17 among Jordanian patients taking clopidogrel

Purpose: To investigate the influence of allelic frequencies of PON1 Q192R, CYP2C19*2 and CYP2C19*17 genetic polymorphisms on the response to clopidogrel among Jordanian patients.Methods: Polymorphisms in CYP2C19 were assessed among 148 patients using PCR-RFLP assay.Results: The CYP2C19*2, CYP2C19*17, and PON1 Q192R allele frequencies were 9.8, 28.72 and 28.7 %, respectively. On the genotyping side, the frequencies of CYP2C19*1/1* and CYP2C19*1/2* were 80.4 and 19.6 %, respectively, but none of the patients had CYP2C19*2/2* genotype. The genotype frequencies CYP2C19*17 were 47.97, 46.62 and 5.41 % for wild-type C-C, heterozygote C-T, and the mutant T-T, respectively. PON1 genotype was 42.7 % for QQ, and 57.8 % for QR. None of the patients had RR genotype.Conclusion: Relative to other populations, the observed allelic frequencies are consistent with the values reported for Caucasian and Middle Eastern populations.Keywords: CYP2C9 polymorphisms, Clopidogrel, Genotype, Allele frequency, PON1 gene

Age-related changes in the phenotype of microglia

Microglia play a central role in immune surveillance and modulation of neuroinflammation as well as playing a role in neurodevelopment. Microglia involve in the development of pathological pain in adults but not early in life. However little detail is known about the changing phenotype of microglia during development. We examined age-related changes in microglia following activation with pathogen- and damage- associated molecular patterns (PAMPs/DAMPs). Microglial cultures were prepared from neonatal postnatal day (P1) and adult (P40) rat brains and spinal cords. Immunocytochemistry, qRT-PCR and functional assays were used to identify age-related differences. Adult microglia display a pro-inflammatory immune profile characterized by significantly increased IL-1β mRNA levels in response to PAMPs and DAMPs. In contrast, IL-1β mRNA in neonatal microglia showed a slight increase after stimulation with DAMPs. Anti-inflammatory gene expression was significantly increased in neonatal microglia relative to adult microglia. Compared to adult microglia, neonatal cells had increased phagocytic activity when unstimulated and following activation with LPS and ATP. Moreover, the nuclear receptor Nurr1 may play a major role in reducing pro-inflammatory signalling and promoting the anti-inflammatory phenotype in neonatal microglia. Nurr1 isoforms are differentially expressed in neonatal and adult microglia, with the Nurr1a isoform being significantly elevated in neonatal cells. Using lentiviral vector-mediated expression of Nurr1 isoforms, we also show that over-expression of TINUR, a splice variant of Nurr1, in neonatal and adult microglia attenuates inflammation by trans-repression the IL-1β expression and trans-activation the IL-10 gene expression following ATP exposure. Together, these data provide evidence for age-related difference in microglial function during postnatal development. In addition, these findings demonstrate insight into the mechanisms by which Nurr1 might act, and suggest potential therapeutic targets for the treatment of neuro-inflammatory diseases

The crosstalk between adenosine A2B receptor and insulin signalling in rat skeletal muscle cells

Diabetes mellitus (DM) is a group of metabolic diseases characterised by hyperglycaemia resulting from defects in insulin secretion, insulin action, or both. Insulin therapy might be affected by specific metabolic enzymes and transporters. There are conflicting reports in the literature on the role of adenosine receptor A2B (AR2B) in skeletal and cardiac muscle glucose metabolism. This study aims to find out if there is an association between AR2B and insulin signalling, especially the metabolic pathways (AKT-GSK). Differentiated L6 cell rat muscle cells were treated with insulin, adenosine agonist NECA, selective AR2B antagonist PSB 603 and combinations between these reagents, the expression of AKT2, GSK3α, and GSK3β were measured by qPCR hydrolysis probe technique. Insulin increases AKT2, GSK3α and GSK3β mRNA expression, while AR2B antagonist inhibits AKT2 GSK3α and GSK3β mRNA expression and combining AR2B antagonist with insulin diminish insulin action and decrease AKT2 GSK3α and GSK3β mRNA expression, which means a strong relationship between AR2B and insulin action. Furthermore AR2B agonist may be a good candidate as an anti-diabetic drug

S1 File -

(XLSX)</p

Comparison between IgE levels of children asthma patients and control group according to GSDMA and GSDMB.

Comparison between IgE levels of children asthma patients and control group according to GSDMA and GSDMB.</p

Comparison between IgE levels of adult asthma patients and control group according to GSDMA and GSDMB.

Comparison between IgE levels of adult asthma patients and control group according to GSDMA and GSDMB.</p

Demographic characteristics of the study population.

Demographic characteristics of the study population.</p

Association of the lung function for the adult asthmatic patients with GSDMA and GSDMB and IgE levels.

Association of the lung function for the adult asthmatic patients with GSDMA and GSDMB and IgE levels.</p

The neuroprotective effect of human primary astrocytes in multiple sclerosis: In vitro model.

Recent studies highlighted the role of astrocytes in neuroinflammatory diseases, particularly multiple sclerosis, interacting closely with other CNS components but also with the immune cells. However, due to the difficulty in obtaining human astrocytes, their role in these pathologies is still unclear. In this study we develop an astrocyte in vitro model to evaluate their role in multiple sclerosis after being treated with CSF isolated from both healthy and MS diagnosed patients. Gene expression and ELISA assays reveal that several pro-inflammatory markers IL-1β, TNF-α and IL-6, were significantly downregulated in astrocytes treated with MS-CSF. In contrast, neurotrophic survival, and growth factors, and GFAP, BDNF, GDNF and VEGF, were markedly elevated upon the same treatment. In summary, this study supports the notion of the astrocyte involvement in MS. The results reveal the neuroprotective role of astrocyte in MS pathogenicity by suppressing excessive inflammation and increasing the expression of tropic factors