AcrySof toric intraocular lens for post-keratoplasty astigmatism

We report a 63-year-old male who had undergone left eye optical penetrating keratoplasty for central leucomatous corneal opacity 10 years earlier. The eye had clear donor graft with residual astigmatism of -6.50 diopter cylinder (DC) at 30°. The patient underwent clear corneal phacoemulsification with implantation of +6.0 D spherical equivalent AcrySof SN60T9 intraocular lens (IOL). Postoperatively, at 10 months, the patient had distance corrected visual acuity of 20/30 with -2.00 DC at 20°. AcrySof toric IOL offers an effective treatment option for post-keratoplasty high corneal astigmatism in patients with cataract.     

Environmental Monitoring and Health Risk Assessment of African Catfish Clarias gariepinus (Burchell, 1822) Cultured in Rural Ponds, India

Water quality monitoring of Clarias gariepinus culture ponds (n?=?27) revealed poor physico-chemical conditions and metal contaminants in fish tissues (n?=?324). Human health risk assessment for some heavy metal contamination delineated low risk in general except for Aluminium (Al), Iron (Fe) and Lead (Pb) which accumulated significantly (p?<?0.05) high in tissues. Health risks values were 6.3?×?10^?3–9.6?×?10^?3 for Al; 3?×?10^?3–9.7?×?10^?3 for Fe and 1.15?×?10^?5–9.3?×?10^?6 for Pb respectively suggesting that contamination of Pb particularly in ponds fed with chicken waste (CW) was posing high risks.     

Redox imbalance in rat tissues exposed with organophosphate pesticides and therapeutic potential of antioxidant vitamins

Organophosphate pesticides are among the most widely used synthetic chemicals for controlling domestic and agricultural pests. Present study was aimed to evaluate the potential of chlorpyrifos, parathion and malathion, to disturb glutathione homeostasis in rat tissues and to find out whether the pre-feeding of antioxidant vitamins has some ameliorating effect on the pesticide-induced alterations. The results showed that these pesticides, alone or in combination, caused decrease in the levels of GSH and the corresponding increase in the levels of GSSG, decreasing the GSH/GSSG ratio. The results also showed NADPH/NADP(+) and NADH/NAD(+) ratios were also decreased in the rat tissues on pesticide exposure. These pesticides, alone or in combination, caused increase in the activities of glutathione reductase and glucose-6-phosphate dehydrogenase in all the rat tissues studied. The findings show that these pesticides generate oxidative stress and prior feeding of mixture of antioxidant vitamins tend to reduce the toxicities of these pesticides.     

Evaluation of recombinant E2 protein-based and whole-virus inactivated candidate vaccines against chikungunya virus

Objectives

With the re-emergence of chikungunya virus (CHIKV) in an explosive form and in the absence of a commercially available vaccine, we aimed to develop candidate vaccines employing recombinant E2 protein or chemically inactivated whole virus.

Design and methods

E2 gene of CHIKV isolate of ECSA genotype was cloned in pET15b vector, expressed and purified (rE2p). The virus was propagated in Vero cell line, purified and inactivated with formalin and BPL individually. Six to eight weeks old female BALB/c mice were immunized intramuscularly with two doses of 10 μg, 20 μg and 50 μg of vaccine formulations with or without adjuvants, 2 weeks apart. The adjuvants evaluated were alum, Mw, CadB (rE2p), alum/Mw (formalin inactivated CHIKV) and alum (BPL-inactivated CHIKV). Humoral immunity was assessed by ELISA and in vitro neutralization test using homologous and heterologous (Asian genotype) strains of CHIKV. Two cohorts of vaccinated mice were challenged separately via intranasal route with homologous virus two and 20 weeks after the 2nd dose. Viral load (CHIKV RNA by real time PCR) was determined in the serum and tissues (muscle, brain, spleen) of the mice challenged with the homologous virus.

Results

Anti-CHIK-antibody titres were dose dependent for all the immunogen formulations. BPL-inactivated vaccines led to the highest ELISA/neutralizing antibody (nAb) titres while alum was the most effective adjuvant. Asian genotype strain could be neutralized by the nAbs. In an adult mouse model, complete protection was offered by the alum-adjuvanted rE2p and both the inactivated vaccines as no virus was detected in the tissues and blood after challenge 2 weeks or 20 weeks-post-2nd dose. However, with rE2p-CadB, very low viremia was recorded on the 2nd day-post-challenge.

Conclusion

Both rE2p and BPL/formalin-inactivated virus are promising candidate vaccines deserving further evaluation.     

High-efficiency transduction of human monocyte-derived dendritic cells by capsid-modified recombinant AAV2 vectors

Phosphorylation of surface-exposed tyrosine residues negatively impacts the transduction efficiency of recombinant AAV2 vectors. Pre-treatment of cells with specific cellular serine/threonine kinase inhibitors also significantly increased the transduction efficiency of AAV2 vectors. We reasoned that site-directed mutagenesis of surface-exposed serine residues might allow the vectors to evade phosphorylation and thus lead to higher transduction efficiency. Each of the 15 surface-exposed serine (S) residues was substituted with valine (V) residues, and the transduction efficiency of three of these mutants, S458V, S492V and S662V, was increased by up to ≈ 20-fold in different cell types. The S662V mutant was efficient in transducing human monocyte-derived dendritic cells (moDCs), a cell type not readily amenable to transduction by the conventional AAV vectors, and did not induce any phenotypic changes in these cells. Recombinant S662V-AAV2 vectors encoding a truncated human telomerase (hTERT) gene were generated and used to stimulate cytotoxic T cells (CTLs) against target cells. S662V-AAV2-hTERT vector-transduced DCs resulted in rapid, specific T-cell clone proliferation and generation of robust CTLs, which led to specific cell lysis of K562 cells. These studies suggest that high-efficiency transduction of moDCs by serine-modified AAV2 vectors is feasible, which supports the potential utility of these vectors for future human DCs vaccine studies.     

Heart-After-Liver Transplantation Attenuates Rejection of Cardiac Allografts in Sensitized Patients

BackgroundIn patients undergoing heart transplantation, significant allosensitization limits access to organs, resulting in longer wait times and high waitlist mortality. Current desensitization strategies are limited in enabling successful transplantation.ObjectivesThe purpose of this study was to describe the cumulative experience of combined heart-liver transplantation using a novel heart-after-liver transplant (HALT) protocol resulting in profound immunologic protection.MethodsReported are the results of a clinical protocol that was instituted to transplant highly sensitized patients requiring combined heart and liver transplantation at a single institution. Patients were dual-organ listed with perceived elevated risk of rejection or markedly prolonged waitlist time due to high levels of allo-antibodies. Detailed immunological data and long-term patient and graft outcomes were obtained.ResultsA total of 7 patients (age 43 ± 7 years, 86% women) with high allosensitization (median calculated panel reactive antibody = 77%) underwent HALT. All had significant, unacceptable donor specific antibodies (DSA) (>4,000 mean fluorescence antibody). Prospective pre-operative flow cytometric T-cell crossmatch was positive in all, and B-cell crossmatch was positive in 5 of 7. After HALT, retrospective crossmatch (B- and T-cell) became negative in all. DSA fell dramatically; at last follow-up, all pre-formed or de novo DSA levels were insignificant at <2,000 mean fluorescence antibody. No patients experienced >1R rejection over a median follow-up of 48 months (interquartile range: 25 to 68 months). There was 1 death due to metastatic cancer and no significant graft dysfunction.ConclusionsA heart-after-liver transplantation protocol enables successful transplantation via near-elimination of DSA and is effective in preventing adverse immunological outcomes in highly sensitized patients listed for combined heart-liver transplantation.     

Protein histidine phosphatase 1 negatively regulates CD4 T cells by inhibiting the K+ channel KCa3.1

The calcium activated K⁺ channel KCa3.1 plays an important role in T lymphocyte Ca²⁺ signaling by helping to maintain a negative membrane potential, which provides an electrochemical gradient to drive Ca²⁺ influx. We previously showed that nucleoside diphosphate kinase beta (NDPK-B), a mammalian histidine kinase, is required for KCa3.1 channel activation in human CD4 T lymphocytes. We now show that the mammalian protein histidine phosphatase (PHPT-1) directly binds and inhibits KCa3.1 by dephosphorylating histidine 358 on KCa3.1. Overexpression of wild-type, but not a phosphatase dead, PHPT-1 inhibited KCa3.1 channel activity. Decreased expression of PHPT-1 by siRNA in human CD4 T cells resulted in an increase in KCa3.1 channel activity and increased Ca²⁺ influx and proliferation after T cell receptor (TCR) activation, indicating that endogenous PHPT-1 functions to negatively regulate CD4 T cells. Our findings provide a previously unrecognized example of a mammalian histidine phosphatase negatively regulating TCR signaling and are one of the few examples of histidine phosphorylation/dephosphorylation influencing a biological process in mammals.