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Reconditioned monocytes are immunomodulatory and regulate inflammatory environment in sepsis

Sun, 01 Jan 2023 00:00:00 GMT

Title: Reconditioned monocytes are immunomodulatory and regulate inflammatory environment in sepsis Authors: Jain, Kshama; Mohan, K Varsha; Roy, Gargi; Sinha, Prakriti; ayaraman, Vignesh J; Kiran; Yadav, Ajit Singh; Phasalkar, Akshay; Deepanshu; Pokhrel, Anupa; Perumal, Nagarajan; Sinha, Nitin; Chaudhary, Kiran; Upadhyay, Pramod Abstract: Sepsis is caused by dysregulated immune response to severe infection and hyper inflammation plays a central role in worsening the disease. The immunomodulatory properties of mesenchymal stem cells (MSCs) have been evaluated as a therapeutic candidate for sepsis. Reconditioned monocytes (RM), generated from healthy human peripheral blood mononuclear cells (PBMCs) exhibit both macrophage and MSCs-like properties. RM were administered at different stages of sepsis in a mouse model. It reduced serum levels of IL6, MCP-1, IL-10, improved hypothermia, increased survival, and recovery from 0 to 66% when combined with antibiotics in the mouse model. The reduced human leucocyte antigen DR molecules expression on RM enables their co-culture with PBMCs of sepsis patients which resulted in reduced ROS production, and up-regulated TGF-β while down-regulating IL6, IL8, and IL-10 in-vitro. RM are potentially immunomodulatory, enhance survival in sepsis mouse model and modulate inflammatory behaviour of sepsis patient's PBMCs.

Transplantation of retinal pigment epithelium and photoreceptors generated concomitantly via small molecule-mediated differentiation rescues visual function in rodent models of retinal degeneration

Fri, 01 Jan 2021 00:00:00 GMT

Title: Transplantation of retinal pigment epithelium and photoreceptors generated concomitantly via small molecule-mediated differentiation rescues visual function in rodent models of retinal degeneration Authors: Harshini Surendran, Harshini; Nandakumar, Swapna; Reddy K, Vijay Bhaskar; Stoddard, Jonathan; Mohan K, Varsha; Upadhyay, Pramod K; McGill, Trevor J; Pal, Rajarshi Abstract: Background: Age-related macular degeneration (AMD) is a result of degeneration/damage of the retinal pigment epithelium (RPE) while retinitis pigmentosa (RP), an inherited early-onset disease, results from premature loss of photoreceptors. A promising therapeutic approach for both is the replacement of lost/damaged cells with human induced pluripotent stem cell (hiPSC)-derived retinal cells. Methods: The aim of this study was to investigate the in vivo functionality of RPE and photoreceptor progenitor (PRP) cells derived from a clinical-grade hiPSC line through a unified protocol. De novo-generated RPE and PRP were characterized extensively to validate their identity, purity, and potency. Results: RPE expressed tight junction proteins, showed pigmentation and ciliation, and secreted polarization-related factors vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF). PRP expressed neural retina proteins and cone and rod markers, and responded to KCl-induced polarization. Transcriptomic analysis demonstrated an increase in the expression of mature retinal tissue-specific genes coupled with concomitant downregulation of genes from undesired lineages. RPE transplantation rescued visual function in RCS rats shown via optokinetic tracking and photoreceptor rescue. PRP transplantation improved light perception in NOD.SCID-rd1 mice, and positive electroretinography signals indicated functional photoreceptor activity in the host's outer nuclear layer. Graft survival and integration were confirmed using immunohistochemistry, and no animals showed teratoma formation or any kind of ectopic growth in the eye. Conclusions: To our knowledge, this is the first demonstration of a unified, scalable, and GMP-adaptable protocol indicating strong animal efficacy and safety data with hiPSC-derived RPE and PRP cells. These findings provide robust proof-of-principle results for IND-enabling studies to test these potential regenerative cell therapies in patients.

Sun exposure, UV irradiance and serum 25-hydroxycholecalciferol in pregnant women in rural north India

Sun, 01 Jan 2017 00:00:00 GMT

Title: Sun exposure, UV irradiance and serum 25-hydroxycholecalciferol in pregnant women in rural north India Authors: Upadhyay, Pramod; Sudhanshu, Siddhnath; Sahu, Monashish; Rawat, Vinita; Bhatia, Vijayalakshmi Abstract: Objective: To document the effect of season and environmental pollution on UVB irradiance; and to estimate cutaneous vitamin D synthesis in village women in different seasons. Design: Radiant UVB energy was measured by a spectroradiometer in different seasons and, in April and May, on successive days in open areas at the city outskirts, at a crowded inner-city area and the villages of our participants. Clothing, outdoor activity pattern and serum 25-hydroxycholecalciferol (25(OH)D) levels were documented. Setting: Rural north India, latitude 26·8°N. Subjects: Pregnant women (n 139, aged 20–40 years). Results: UVB irradiance ranged from 56 μW/cm2 in January to 470 μW/cm2 in June. Proportion of skin exposed was 18·5% in summer and 9·5% in winter. Mean (SD) daily duration of sun exposure was 3·2 (0·2) h during winter and 2·1 (0·4) h during summer. Cutaneous vitamin D synthesis was estimated to be 19·25 μg (770 IU) during winter and 37·25 μg (1490 IU) during summer. Mean (SD) serum 25(OH)D was 28 (15) nmol/l during winter (92% of participants with <50 nmol/l) and reached 56 (20) nmol/l during late summer (60% with >50 nmol/l). Mean (SD) UVB irradiance at peak summer was significantly higher at the open areas and in the villages than at the inner-city location (340 (45) and 310 (60) v. 250 (50) μW/cm2, P=0·03). Conclusions: In our population, at latitude 26·8°N, poor skin exposure is a limiting factor in all seasons. During winter, low UVB radiation energy also contributes. Particulate pollution limits UVB irradiance. Vitamin D supplementation during winter may be necessary.

Autologous NeoHep Derived from Chronic Hepatitis B Virus Patients' Blood Monocytes by Upregulation of c-MET Signaling

Sun, 01 Jan 2017 00:00:00 GMT

Title: Autologous NeoHep Derived from Chronic Hepatitis B Virus Patients' Blood Monocytes by Upregulation of c-MET Signaling Authors: Upadhyay, Pramod Abstract: In view of the escalating need for autologous cell-based therapy for treatment of liver diseases, a novel candidate has been explored in the present study. The monocytes isolated from hepatitis B surface antigen (HBsAg) nucleic acid test (NAT)-positive (HNP) blood were differentiated to hepatocyte-like cells (NeoHep) in vitro by a two-step culture procedure. The excess neutrophils present in HNP blood were removed before setting up the culture. In the first step of culture, apoptotic cells were depleted and genes involved in hypoxia were induced, which was followed by the upregulation of genes involved in the c-MET signaling pathway in the second step. The NeoHep were void of hepatitis B virus and showed expression of albumin, connexin 32, hepatocyte nuclear factor 4-α, and functions such as albumin secretion and cytochrome P450 enzyme-mediated detoxification of xenobiotics. The engraftment of NeoHep derived from HBsAg-NAT-positive blood monocytes in partially hepatectomized NOD.CB17-Prkdcscid /J mice liver and the subsequent secretion of human albumin and clotting factor VII activity in serum make NeoHep a promising candidate for cell-based therapy.