Correction of ankle and hind foot deformity in Charcot neuroarthropathy using a retrograde hind foot nail—The Kings’ Experience

Background

Corrective fusion for the unstable deformed hind foot and mid foot in Charcot Neuroarthropathy (CN) is quite challenging and is best done in tertiary centres under the supervision of multidisciplinary teams.

Identification and characterization of nematode specific protective epitopes of Brugia malayi TRX towards development of synthetic vaccine construct for lymphatic filariasis

Although multi-epitope vaccines have been evaluated for various diseases, they have not yet been investigated for lymphatic filariasis. Here, we report for the first time identification of two immunodominant B epitopes (TRX_P1 and TRX_P2) from the antioxidant Brugia malayi thioredoxin by studying their immune responses in mice model and human subjects. TRX_P1 was also found to harbor a T epitope recognized by human PBMCs and mice splenocytes. Further, the epitopic peptides were synthesized as a single peptide conjugate (PC1) and their prophylactic efficacy was tested in a murine model of filariasis with L3 larvae. PC1 conferred a significantly high protection (75.14%) (P < 0.0001) compared to control (3.7%) and recombinant TRX (63.03%) (P < 0.018) in experimental filariasis. Our results suggest that multi-epitope vaccines could be a promising strategy in the control of lymphatic filariasis.     

The amino acid metabolism is essential for evading physical plasma-induced tumour cell death

Background Recent studies have emphasised the important role of amino acids in cancer metabolism. Cold physical plasma is an evolving technology employed to target tumour cells by introducing reactive oxygen species (ROS). However, limited understanding is available on the role of metabolic reprogramming in tumour cells fostering or reducing plasma-induced cancer cell death.Methods The utilisation and impact of major metabolic substrates of fatty acid, amino acid and TCA pathways were investigated in several tumour cell lines following plasma exposure by qPCR, immunoblotting and cell death analysis.Results Metabolic substrates were utilised in Panc-1 and HeLa but not in OVCAR3 and SK-MEL-28 cells following plasma treatment. Among the key genes governing these pathways, ASCT2 and SLC3A2 were consistently upregulated in Panc-1, Miapaca2GR, HeLa and MeWo cells. siRNA-mediated knockdown of ASCT2, glutamine depletion and pharmacological inhibition with V9302 sensitised HeLa cells to the plasma-induced cell death. Exogenous supplementation of glutamine, valine or tyrosine led to improved metabolism and viability of tumour cells following plasma treatment.Conclusion These data suggest the amino acid influx driving metabolic reprogramming in tumour cells exposed to physical plasma, governing the extent of cell death. This pathway could be targeted in combination with existing anti-tumour agents.Subject terms: Physics, Cancer metabolism     

Evidence for non‐functional Dickkopf‐1 (DKK‐1) signaling in chronic lymphocytic leukemia (CLL)

Purpose: Wnt signaling was demonstrated to be activated in chronic lymphocytic leukemia (CLL). It is thought to be responsible for the extended survival of CLL cells in vivo. Dickkopf1 (DKK1) is known to antagonize Wnt signaling by direct high‐affinity binding to the extracellular domain of WNT coreceptor lipoprotein receptor‐related protein 6 (LRP6). The purpose of this study was to investigate the effect of DKK1 in B‐CLL cells in vitro. Methods: Expression of DKK1 was estimated by Western blot and real‐time PCR. B cells from patients with CLL and healthy donors were incubated with recombinant DKK1. Survival was measured by flow cytometry. Primers for real‐time PCR were designed for extracellular domain of LRP6, responsible for DKK1 binding, and the intracellular region, essential for inhibiting GSK3 β. Results: Healthy and CLL cells express equivalent mRNA levels of DKK1 and LRP6. After treatment of CLL cells with recombinant DKK1 (1 μg/mL) for 3 h, there was no change in the levels of phosphorylated β‐catenin and total β‐catenin. Healthy B cells proved to have significantly higher levels of extracellular, DKK1 binding domain of LRP6. We estimated that in CLL cells every 6th LRP6 receptor is lacking the extracellular domain. Discussion: For the first time we show the expression of DKK1 in CLL cells. Unlike in similar tumors, the addition of DKK1 to culture of CLL cells does not inactivate WNT pathway. The reason for this could be the absence of the binding domain of LRP6. On the other hand, a truncated LRP6 without extracellular DKK1 binding domain could lead to an uncontrollable activation of WNT signaling.     

Immune responses of B. malayi thioredoxin (TRX) and venom allergen homologue (VAH) chimeric multiple antigen for lymphatic filariasis

Although multiple vaccine strategy for lymphatic filariasis has provided tremendous hope, the choice of antigens used in combination has determined its success in the previous studies. Multiple antigens comprising key vaccine candidates from different life cycle stages would provide a promising strategy if the antigenic combination is chosen by careful screening. In order to analyze one such combination, we have used a chimeric construct carrying the well studied B. malayi antigens thioredoxin (BmTRX) and venom allergen homologue (BmVAH) as a fusion protein (TV) and evaluated its immune responses in mice model. The efficacy of fusion protein vaccine was explored in comparison with the single antigen vaccines and their cocktail. In mice, TV induced significantly high antibody titer of 1,28,000 compared to cocktail vaccine TRX+VAH (50,000) and single antigen vaccine TRX (16,000) or VAH (50,000). Furthermore, TV elicited higher level of cellular proliferative response together with elevated levels of IFN-γ, IL-4 and IL-5 indicating a Th1/Th2 balanced response. The isotype antibody profile showed significantly high level of IgG1 and IgG2b confirming the balanced response elicited by TV. Immunization with TV antigen induced high levels of both humoral and cellular immune responses compared to either cocktail or antigen given alone. The result suggests that TV is highly immunogenic in mice and hence the combination needs to be evaluated for its prophylactic potential.     

Need and Feasibility of a Transition Clinic for Adolescents with Chronic Illness: A Qualitative Study

To assess the need and feasibility of a Transition Clinic in the care of adolescents with chronic illness. A qualitative study, piloted by a questionnaire-based survey, followed by Focus Group Discussions was conducted in the Department of Pediatrics in a government teaching hospital in Kerala. The participants were adolescents with chronic disease, viz., renal disease, diabetes, HIV/AIDS, cancer survivors with Hepatitis B, parents of renal patients, pediatricians and adult specialists. Twenty patients were administered a structured, rated, 18-item questionnaire regarding their attitude and preparedness for transfer to adult departments. The need and feasibility of a Transition Clinic was discussed by 7 focus groups viz., pediatricians, adult physicians, 4 patient groups and 1 parent group. Thematic analysis of data was done. Except among pediatricians, awareness of the concept of Transition Clinic was low. All participants agreed that abrupt transfer of care hinders treatment and that there is a definite need for a Transition Clinic. Pediatricians discussed the complex needs of adolescents and limitations of their care. Adult specialist physicians described their inadequacy to handle adolescent problems and difficulties posed by large volumes of patients. The adolescents and young adults with chronic illness discussed their reluctance to shift to the adult departments. Parents voiced their concerns about the future of their children. All groups gave constructive suggestions for conduct of the clinic and smooth transition. There is a definite need for a Transition Clinic in the management of adolescents with chronic illness to ensure smooth transfer of care. Introducing such clinics in the existing health framework is feasible using a multidisciplinary approach.