Thymic and Peripheral T-cell Polarization in an Experimental Model of Russell's Viper Venom-induced Acute Kidney Injury.

Venom pathology is not restricted to the direct toxic effects of venom. Immunoinflammatory alteration as the etiology of snake venom-induced acute kidney injury (SAKI) is a less trodden path toward the development of alternative therapeutic approach. In the present study, we have associated the crest of renal damage stage to the immunological alteration, as reflected in thymic and peripheral T cell polarization in the murine model of SAKI. Renal injury in mice was confirmed from significant dysuresis and adversely altered biochemical renal markers. Histopathological alterations, as revealed by marked tubular and glomerular damage, reaffirmed kidney injury. SAKI is accompanied by significant inflammatory changes as indicated by neutrophilic leucocytosis, increased neutrophil to lymphocyte ratio and plasma CRP levels. Thymic immunophenotyping revealed significantly increased CD8+ cytotoxic T cell, and CD25+ both single positive population (p = .017-0.010) and CD44-CD25+ double negative population (DN3) (p = .002) accompanied by an insignificantly reduced CD4+ helper T cells (p = .451). Peripheral immunophenotyping revealed similar pattern as indicated by reduced helper T cells (p = .002) associated with significantly elevated cytotoxic T cells (p = .009) and CD25+ subset of both helper (p = .002) and cytotoxic (p = .024) T cells. The IL-10+ subset of both CD25+ and CD25- T cells were also found to be significantly elevated in the SAKI group (p ≤ 0.020) suggesting an immunosuppressive phenotype in SAKI. It can be concluded that T cells responds to venom-induced renal injury particularly through IL-10+ reparative phenotypes which are known for their immunosuppressive and anti-inflammatory activity.

On-chip estimation of hematocrit level for diagnosing anemic conditions by Impedimetric techniques.

An on-chip device has been fabricated on Si platform for precise estimation of the hematocrit (Hct) level of human blood with rapid turn out. Impedance/capacitance spectroscopy and current-voltage (I-V) measurements have been employed to observe the variation of electrical parameters of erythrocyte suspensions with varying Hct level. Experimentally obtained values of capacitance, impedance and conductance with Hct level suggests a linear variation. Current-time measurement has also been performed to ensure the susceptibility of red blood cells under a fixed external electric field for certain duration of time. The online real time sample analyzes have also been performed by the device connected with an embedded electronic circuit interfaced with a laptop through appropriate software. This has enabled the development of a novel Si-chip based compact point-of-care (POC) diagnosis system for Hct level estimation by measuring the dielectric/capacitive variation of an erythrocyte cell suspension. The relevant performance parameters of such a compact system including range, resolution, limit of detection and throughput have also been evaluated.

Sustained release of ketorolac tromethamine from poloxamer 407/cellulose nanofibrils graft nanocollagen based ophthalmic formulations.

There has been extensive utilization of poloxamer 407 (PM) for the delivery of various ophthalmic drugs aimed at efficient ophthalmic drug delivery approach for longer precorneal residence time along with acceptable bioavailability of drugs. We have studied the effect of nanocellulose grafted collagen (CGC) on the performance of in situ gels based on PM for the controlled in vitro release of Ketorolac Tromethamine (KT). CGC has shown great influence evident by the reduction in PM critical gelation concentration, increased gel strength, and prolonged the release of loaded drugs compared with the virgin PM gel. The engineered nanocomposite formulations established an anomalous diffusion mechanism along with a Fickian diffusion controlled drug release for 1.5 & 1.75 w/v% CGC reinforced PM. Hence, the synthesized in situ nanocomposites are potential candidates for ophthalmic drug delivery system.

Effect of cellulose nanocrystals on the performance of drug loaded in situ gelling thermo-responsive ophthalmic formulations.

Triblock poloxamer copolymer (PM) has been extensively utilized to deliver various ophthalmic pharmaceutical compounds. The aim of efficient ophthalmic drug delivery strategy is to attain the longer precorneal resident time and good bioavailability of drugs. In this pursuit, the influence of cellulose nanocrystals (CNC) on the in situ gelation behavior of PM and in vitro release of pilocarpine hydrochloride from the nanocomposites formulations was studied. The critical concentration of gelation of PM being 18% (wt/v) was dropped to 16.6% (wt/v) by the addition of a very low percentage of CNC. The reinforcing nature of CNC via H-bonding in the in situ nanocomposite gel also led to an increase in gel strength along with the sustained release of loaded drugs when compared with the pure PM gel. All formulations revealed that the drug release mechanism is controlled by the Fickian diffusion. Thus, the CNC has a significant effect on the gelation behavior, gel strength, and drug release kinetics of PM-CNC formulations.

Hematological alterations and splenic T lymphocyte polarization at the crest of snake venom induced acute kidney injury in adult male mice.

Snake venom induced acute kidney injury (SAKI) is of great clinical relevance in tropical countries. Involvement of T cell, a key mediator of AKI and its remission, is least explored in SAKI. In the present study the in vivo hematological alterations and associated splenic T cell polarization is probed in order to investigate the immune response at the crest of Russell's viper venom (RVV) induced AKI in experimental murine model. Based on a dose and time kinetic study intra muscular injection dose of 20 µg RVV/100 gm body weight of mice and incubation period of 60 h was selected for induction of SAKI. Renal involvement in SAKI group was confirmed from oliguria, significantly elevated urinary microprotein (p < 0.001), decreased urinary creatinine (p = 0.003) and creatinine clearance (p < 0.001) compared to control. Hematological analyses revealed a significant neutrophilic leukocytosis (p < 0.001) associated with a reduced lymphocyte percentage (p < 0.001) favoring a state of acute inflammation in SAKI group. Immunophenotyping study of splenocytes showed a significant decrease in CD4+/CD8+ ratio (p < 0.001) with a significant increase in regulatory (CD25+FoxP3+) helper and cytotoxic subset of T cell (p < 0.001). Significant increase in IL-10+ regulatory helper and cytotoxic T cell (p < 0.001) further confirmed the internal milieu favoring immunosuppression. Apart from these the CD25-FoxP3+ reservoir regulatory T cells were also found to be significantly elevated in SAKI group compared to that of control (p < 0.001). Taken together, the results of the present study clearly indicated a state of acute inflammation and splenic T cell polarization towards regulatory subset at the crest of SAKI.