Mechanisms of ß-adrenergic receptors agonists in mediating pro and anti-apoptotic pathways in hyperglycemic Müller cells.

BACKGROUND: The current study aimed to investigate the stimulatory effect of beta-adrenergic receptors (ß-ARs) on brain derived neurotropic factor (BDNF) and cAMP response element binding protein (CREB). METHODS: Human Müller cells were cultured in low and high glucose conditions. Cells were treated with xamoterol (selective agonist for ß1-AR), salmeterol (selective agonist for ß2-AR), isoproterenol (ß-ARs agonist) and propranolol (ß-ARs antagonist), at 20 µM concentration for 24 h. Western Blotting assay was performed for the gene expression analysis. DNA damage was evaluated by TUNEL assay. DCFH-DA assay was used to check the level of reactive oxygen species (ROS). Cytochrome C release was measured by ELISA. RESULTS: Xamoterol, salmeterol and isoproterenol showed no effect on Caspase-8 but it reduced the apoptosis and increased the expression of BDNF in Müller cells. A significant change in the expression of caspase-3 was observed in cells treated with xamoterol and salmeterol as compared to isoproterenol. Xamoterol, salmeterol and isoproterenol significantly decreased the reactive oxygen species (ROS) when treated for 24 hours. Glucose-induced cytochrome c release was disrupted in Müller cells. CONCLUSION: ß-ARs, stimulated by agonist play a protective role in hyperglycemic Müller cells, with the suppression of glucose-induced caspase-3 and cytochrome c release. B-Ars may directly mediate the gene expression of BDNF.

Hippophae rhamnoides mediate gene expression profiles against keratinocytes infection of Staphylococcus aureus.

Staphylococcus aureus causes a wide range of skin diseases such as bacterial keratitis, follicles, psoriasis, cellulitis and atopic dermatitis. This study aims to investigate the S. aureus mediated molecular modulation, and the effect of HR in reversing the deleterious impact of S. aureus in keratinocytes. Human keratinocyte (HaCaT) cells were cultured in DMEM, supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. Subcultures were divided into three flasks: control with no S. aureus and extract (C), S. aureus infected (SA) and S. aureus infected cells and extract (SE). RNA was isolated and microarray analysis was performed. The data was annotated using GO functional analysis and DAVID functional annotation. For each comparison group, significant probes were filtered out at significant cut off by fold change (P < 0.05 and/or > twofold change). For SA vs control, SE vs control, and SE vs SA, 204, 9369, 9900 probes were filtered respectively. In SA vs control, TNF signaling, NOD-like receptor and chemokine receptor signaling pathways were upregulated with key genes such as CCL2, CCL20 and BIRC3. The SE vs SA, showed significant expression variations of a number of important genes. Molecular pathways associated with ILs, TNFs, TGFs, IFNs, FGFs, MAPKs, MMPs, caspases and Wnts were either up regulated or downregulated. This effect was reversed by the extract, possibly through downregulating various proinflammatory cytokines and apoptotic pathways. Our study reveals that S. aureus inserts a negative impact on the regulation of various key genes which is apparently reversed by the HR extract.

Beta Adrenergic Receptors Stimulation Attenuates Phosphorylation of NF-κB and IκBα in Hyperglycemic Endothelial Cells.

BACKGROUND/AIMS: NF-κB induces transcription of a number of genes, associated with inflammation and apoptosis. In this study, we have investigated the effect of ß-adrenergic receptor stimulation on NF-κB and IκBα in HUVECs. METHODS: Human umbilical vein endothelial cells (HUVECs) were cultured in high and low glucose concentrations. All HUVECs were treated with different concentrations of isoproterenol and propranolol for different time periods. The analytical procedures consisted of Western Blot, ELISA, DCFH-DA and TUNEL assays. RESULTS: Isoproterenol (agonist of a beta-adrenergic receptor) significantly reduced phosphorylation at Ser-536 of NF-κB; and Ser-32 and Ser-36 of IκBα in hyperglycemic HUVECs. Isoproterenol also significantly reduced apoptosis and ROS generation. No effect of IκBα was observed on Tyr-42 phosphorylation. The effect of isoproterenol was reversed by the antagonist propranolol. We also checked if NF-κB inhibitor MG132 causes any change at the level of apoptosis. However, we observed an almost similar effect. CONCLUSION: Given data demonstrates that beta-adrenergic receptors stimulation has a protective effect on HUVECs that might be occuring via NF-κß and IκBα pathway.

Seasonal changes in infrapopulations of Diplozoon kashmirensis Kaw, 1950 (Monogenea: Diplozoidae) along a eutrophic gradient.

Seasonal population dynamics of the monogenean, Diplozoon kashmirensis Kaw, on the gills of two cyprinid fish species, Schizothorax niger Heckel and Carassius carassius (Linnaeus), was investigated in three limnologically distinct trophic habitats located along the flood plain of River Jhelum in Kashmir from June 2006 to May 2008. The parasite infrapopulations exhibited a marked seasonal regime in infestation pattern as the infection indices increased to a much higher plateau during summer season at all the lakes, while the lows were recorded in winter. The heterogeneity in infection pattern indicates that water temperature is an important determinant of the seasonality of infrapopulations at all the localities. Furthermore, the results of our work clearly indicate that the parasite infrapopulations increased proportionally with eutrophication level and, as such, the highly eutrophic habitat, Anchar Lake, was significantly more favourable for parasite infrapopulations than the less eutrophic ones. However, the lakes presented no significant interlake differences in water temperature. Therefore, we could argue that interlake differences in the infestation pattern of parasite can be safely attributed to respective water quality in the lakes rather than water temperature. We propose that infrapopulations of the diplozoid studied herein do respond to differences in water quality of lakes and, thus, could qualify as simple and reliable indicator species in short-term comparative assays by lake managers.

Metacercariae of Clinostomum schizothoraxi Kaw, 1950 (Digenea: Clinostomatidae) in Carassius carassius (Linnaeus) under different environmental conditions.

This report addresses the possible impacts of local habitat characteristics on the metacercariae of Clinostomum schizothoraxi Kaw, 1950 infecting crucian carp, Carassius carassius (Linnaeus), from three lakes in Kashmir. The lakes chosen encompass an extremely wide gradient in trophic status--Manasbal (less polluted and mesotrophic), Dal (moderately polluted and eutrophic) and Anchar (strongly polluted and hypertrophic). The results indicate that infrapopulation-level descriptors of abundance of C. schizothoraxi in fish differed between the three lakes and revealed that the infection levels were greater at Anchar Lake. Furthermore, the prevalence, mean abundance and mean intensity of infection also changed during the period of investigation along the seasonal gradient of water temperature. The fact that the abundance patterns of the parasite differed in different populations of the same host provided a unique opportunity for a comparative study on the temporal variations in infection patterns attained during different seasons. In particular, the lake environments showed a high degree of variability in the density profiles of mollusc intermediate hosts and, thus, the differences in the infection status of crucian carp were derived on the basis of intermediate host population size which, in turn, was influenced by the trophic characteristics of the three lakes.

Helminth communities of fish as ecological indicators of lake health.

This paper deals largely with the dynamics and changes in the helminth parasite communities of fish along the trophic gradient of lakes. The use of parasitological community data as a bioindicator of environmental health underlines the need to study parasite communities at comparable localities with known pollution levels. The comparison of the conditions in different habitats might be helpful to differentiate between normal fluctuations in ambient conditions and pollution-mediated effects. Therefore, the present study was designed to examine the community structure of parasites in snow trout (Schizothorax niger Heckel) inhabiting 3 lakes of contrasting trophic status in Kashmir. The idea of selecting the lakes, namely Anchar (strongly hypereutrophic), Dal (eutrophic) and Manasbal (mesotrophic) for this study was intentional as they depict different trophic gradients and exhibit the desirable pattern which was a prerequisite for this study. The findings presented in this article suggest an apparent lake-wise gradient in community structure, as the increase in trematode and cestode infections in Anchar was markedly greater, to levels clearly distinguishable from those in the other two water bodies. We conclude that human-induced eutrophication of lakes modifies the parasite community at component level and community-level studies on parasites may provide information on health status of lakes.