The complement cascade is a central component of innate immunity which plays a critical role in brain inflammation. Complement C3a receptor (C3aR) is a key mediator of post-ischemic cerebral injury, and pharmacological antagonism of the C3a receptor is neuroprotective in stroke. Cerebral ischemia injures brain endothelial cells, causing blood brain barrier (BBB) disruption which further exacerbates ischemic neuronal injury. In this study, we used an in vitro model of ischemia (oxygen glucose deprivation; OGD) to investigate the protective effect of a C3aR antagonist (C3aRA, SB290157) on brain endothelial cells (bEnd.3). Following 24 hours of reperfusion, OGD-induced cell death was assessed by TUNEL and Caspase-3 staining. Western blot and immunocytochemistry were utilized to demonstrate that OGD upregulates inflammatory, oxidative stress and antioxidant markers (ICAM-1, Cox-2, Nox-2 and MnSOD) in endothelial cells and that C3aRA treatment significantly attenuate these markers. We also found that C3aRA administration restored the expression level of the tight junction protein occludin in endothelial cells following OGD. Interestingly, OGD/reperfusion injury increased the phosphorylation of ERK1/2 and C3aR inhibition significantly reduced the activation of ERK suggesting that endothelial C3aR may act via ERK signaling. Furthermore, exogenous C3a administration stimulates these same inflammatory mechanisms both with and without OGD, and C3aRA suppresses these C3a-mediated responses, supporting an antagonist role for C3aRA. Based on these results, we conclude that C3aRA administration attenuates inflammation, oxidative stress, ERK activation, and protects brain endothelial cells following experimental brain ischemia.
Blood-Brain Barrier ; Blotting, Western ; Brain Ischemia ; Brain ; Caspase 3 ; Cell Death ; Complement C3a ; Complement System Proteins ; Encephalitis ; Endothelial Cells ; Glucose ; Immunity, Innate ; Immunohistochemistry ; In Situ Nick-End Labeling ; In Vitro Techniques ; Inflammation ; Ischemia ; Neurons ; Occludin ; Oxidative Stress ; Phosphorylation ; Reperfusion ; Stroke ; Tight Junctions

Objective: To evaluate and compare the antioxidant activity, total phenolic contents (TPCs) and in vitro antidiabetic activity of various pits extracts obtained from five Omani date cultivars. Methods: Sun-dried mature fruits of five Omani date varieties, namely, Fardh, Naghal, Khalas, Khinazi and Khasab were purchased from the local market in Muscat, Oman in the month of September 2014. Four seed extracts viz. water, ethanol, methanol and acetone were prepared for each date variety and their antioxidant activities were investigated by 1,1-diphenyl-2-picrylhydrazyl, hydrogen peroxide scavenging method and reducing power assay method, respectively. In vitro antidiabetic activity of the date pit extracts was evaluated by measuring their inhibitory effect on α-glucosidase and α-amylase level. TPCs were also quantified colorimetrically. Results: The results indicated that TPC of date seeds was solvent dependent. Acetone, ethanol and methanol were found to be significantly better solvents than water in extracting phenolic compounds from the date seeds. Pit extracts exhibited moderate to good in vitro antioxidant activity and increased reducing power. Among all date pit extracts, water extract exhibited significant in vitro antidiabetic activity in comparison to standard drug, acarbose. Conclusions: The present study confirms that disposed waste of Omani dates is a rich source of dietary antioxidant because of its high TPC. The pits due to their inhibitory effects on α-glucosidase and α-amylase level could be used as a monotherapy along with an appropriate diabetic diet and exercise or might be in conjunction with antidiabetic therapy to manage and prevent progression of diabetes.

Aims: This study was aimed to screen indigenous medicinal plants for their antibacterial potential against methicillin-resistant Staphylococcus aureus (MRSA). Methodology and results: Three indigenous plants (Nigella sativa, Zingiber officinale and Calotropis procera) and thymoquinone were screened for antibacterial activity against MRSA, isolated from septic wounds of patients admitted to Mayo Hospital Lahore, Pakistan. Isolated bacteria were screened for methicillin and cefoxitin resistance by the Kirby-Bauer method, followed by mecA gene-specific polymerase chain reaction (PCR). Confirmed MRSA was processed for antibacterial activity of plant extracts and thymoquinone followed by cytotoxicity assay of plant extract having least minimum inhibitory concentration (MIC) value. Out of total samples (n=100), S. aureus (29%), MRSA (26%) and vancomycin-resistant S. aureus (VRSA) (21.7%) isolates were recovered based on morphology, biochemical profile and antibiotic susceptibility testing. Nigella sativa showed the highest antibacterial activity (10.06 ± 6.53 mm) against MRSA followed by Z. officinale (4.06 ± 3.72 mm) and C. procera (3.65 ± 3.33 mm) in comparison to standard thymoquinone (17.93 ± 10.14 mm). The least MIC value recorded was for Z. officinale at 36.89 ± 3.75 μg/mL. Zingiber officinale was the most effective antibacterial agent, followed by N. sativa and C. procera and non-toxic for eukaryotic cells at all tested concentrations (1500 μg/mL to 2.92 μg/mL). Conclusion, significance and impact of study It was concluded that Z. officinale may be used as an effective alternative for treating septic wound infection in local or topical preparations. As pathogenic S. aureus is becoming life-threatening among antibiotic-resistant bacteria and traditional plants are in used for centuries to treat septic wound infections.
Plants, Medicinal ; Methicillin-Resistant Staphylococcus aureus--isolation & ; purification ;

Objective To summarize the precise association between pulmonary tuberculosis (PTB) and P2x7 A1513C gene polymorphism. Methods PubMed and Google Scholar web-databases were searched for the studies reporting the association of P2x7 A1513C polymorphism and PTB risk. A meta-analysis was performed for the selected case–control studies and pooled odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated for all the genetic models. Results Eleven studies comprising 2 678 controls and 2 113 PTB cases were included in this meta-analysis. We observed overall no significant risk in all the five genetic models. When stratified population by the ethnicity, Caucasian population failed to show any risk of PTB in all the genetics models. In Asian ethnicity, variant allele (C vs. A: P = 0.001; OR = 1.375, 95% CI = 1.159–1.632) and heterozygous genotype (AC vs. AA: P = 0.001; OR = 1.570, 95% CI = 1.269–1.944) demonstrated significant increased risk of PTB. Likewise, recessive genetic model (CC + AC vs. AA: P = 0.001; OR = 1.540, 95% CI = 1.255–1.890) also demonstrated increased risk of PTB in Asians. Conclusions Our meta-analysis did not suggest the association of P2x7 A1513C polymorphism with PTB risk in overall or separately in Caucasian population. However, it plays a significant risk factor for predisposing PTB in Asians. Future larger sample and expression studies are needed to validate this association.