Repurposing alagebrium for diabetic foot ulcer healing: Impact on AGEs/NFκB/NOX1 signaling.

BACKGROUND: Diabetic foot ulcer (DFU) is a common diabetic complication associated with disability and reduced quality of life. Available therapeutics are not sufficient to combat the spread of DFU. Here we aim to investigate the impact of alagebrium, an advanced glycation end product (AGE)-crosslink breaker, on the healing of DFU. METHODS: Diabetes was induced in Wistar rats by STZ, and after four weeks, wound was induced on the foot. Alagebrium (10 mg/kg) was administered orally for 14 days, and wound size was measured every 3 days. Behavioral tests i.e., hot plate and footprint tests, were performed to assess sensory function and gait. Blood was collected to assess HbA1c, serum AGEs, MDA and NOX1. Tissue was collected to assess histological changes and expression of NF-κB, iNOS, TNF-α, VEGF and EGF. In a subsequent set of experiments with similar design, alagebrium was applied topically as a film-forming gel. RESULTS: Systemic alagebrium treatment accelerated the healing of diabetic wound, improved sensory functions and gait, and ameliorated histological changes. It also reduced serum levels of AGEs, MDA and NOX1, and the tissue expression of NF-κB, iNOS, TNF-α, and increased VEGF and EGF in diabetic rats. Topical alagebrium led to similar beneficial effects i.e., accelerated diabetic wound healing, improved wound histological changes, reduced expression of NF-κB and iNOS and increased VEGF. CONCLUSIONS: Our findings suggest repurposing of alagebrium for the management of DFU to accelerate the healing process and improve the clinical outcomes in diabetic patients.

The controverted therapeutic efficacy of Allium sativum and Artemisia herba-alba extracts on Cryptosporidium-infected mice.

INTRODUCTION: Cryptosporidiosis has become an issue of great interest being life-threatening among immunocompromised hosts worldwide. This study explored the curative effect of Allium sativum (garlic) and Artemisia herba-alba ethanolic extract versus Nitazoxanide drug on both immunocompetent and immunosuppressed-Cryptosporidium experimentally-infected mice. METHODOLOGY: One hundred male Swiss albino mice were divided into the following groups: (GI) non-infected non-treated group, (GII) infected non-treated group, (GIII) garlic treated group, (GIV) A. herba-alba treated group, (GV) Nitazoxanide treated group, each group subdivided into two subgroups (a) Immunocompetent, (b) immunosuppressed. The assessment was performed by parasitological counting of fecal oocysts, histological examination of intestinal tissue, immunological detection of interferon-gamma levels in mice sera, and ultrastructural study by transmission electron microscopy. RESULTS: Garlic and A. herbal-alba extracts showed a decrease in the mean oocyst counts through all days of follow-up. This was associated with significant up-regulation of interferon-gamma cytokine levels in serum and histological improvement in intestinal tissues of mice compared to control groups and the results were confirmed by transmission electron microscopy. The highest efficacy was obtained by garlic, then by A. herbal-alba extracts followed by Nitazoxanide treated group; where the immunocompetent groups showed better improvement than immunosuppressed ones. CONCLUSIONS: Garlic has a perfect effect as a promising therapeutic agent against Cryptosporidiosis and therefore validates their traditional use in parasitic infections. Accordingly, it may offer a good option for cryptosporidium treatment in immunocompromised patients. They could be used as a natural safe product for the preparation of a new therapeutic agent.

The potential therapeutic effect of Nigella sativa and Zingiber officinale extracts versus Nitazoxanide drug against experimentally induced cryptosporidiosis in laboratory mice.

In this study, the potential anti-cryptosporidial effect of Nigella sativa (black seeds) and Zingiber officinale (ginger) alcoholic extracts versus Nitazoxanide (NTZ) medication was investigated in immunosuppressed (IS) laboratory mice. Parasitological, histopathological studies were used to assess their therapeutic efficacy. Serum level and tissue expression percentage of IFN-γ was also used. Nigella extract succeeded to reduce the mean oocyst counts in the feces of immunosuppressed mice followed by NTZ. Ginger-treated ones showed the lowest reduction percentage. Nigella sativa showed the best results in terms of restoring the normal architecture of ileal epithelium in histopathological sections stained with H&E. NTZ treatment sub-groups showed mild improvement, followed by ginger-treated mice, which showed a slight improvement in small intestine microenvironment. A significant substantial rise in serum and intestinal tissue IFN-γ cytokine levels were recorded in Nigella subgroups compared to those of NTZ and ginger respectively. According to our findings Nigella sativa outperformed Nitazoxanide in terms of anti-cryptosporidial effectiveness and regeneration characteristics revealing a promising medication. When compared to the commonly used Nitazoxanide medication or Nigella extracts, the outcomes of ginger extract were suboptimal.

Defining the Optimal FVIII Transgene for Placental Cell-Based Gene Therapy to Treat Hemophilia A.

The delivery of factor VIII (FVIII) through gene and/or cellular platforms has emerged as a promising hemophilia A treatment. Herein, we investigated the suitability of human placental cells (PLCs) as delivery vehicles for FVIII and determined an optimal FVIII transgene to produce/secrete therapeutic FVIII levels from these cells. Using three PLC cell banks we demonstrated that PLCs constitutively secreted low levels of FVIII, suggesting their suitability as a transgenic FVIII production platform. Furthermore, PLCs significantly increased FVIII secretion after transduction with a lentiviral vector (LV) encoding a myeloid codon-optimized bioengineered FVIII containing high-expression elements from porcine FVIII. Importantly, transduced PLCs did not upregulate cellular stress or innate immunity molecules, demonstrating that after transduction and FVIII production/secretion, PLCs retained low immunogenicity and cell stress. When LV encoding five different bioengineered FVIII transgenes were compared for transduction efficiency, FVIII production, and secretion, data showed that PLCs transduced with LV encoding hybrid human/porcine FVIII transgenes secreted substantially higher levels of FVIII than did LV encoding B domain-deleted human FVIII. In addition, data showed that in PLCs, myeloid codon optimization is needed to increase FVIII secretion to therapeutic levels. These studies have identified an optimal combination of FVIII transgene and cell source to achieve clinically meaningful levels of secreted FVIII.

Soluble Fas affects erythropoiesis in vitro and acts as a potential predictor of erythropoiesis-stimulating agent therapy in patients with chronic kidney disease.

Serum soluble Fas (sFas) levels are associated with erythropoietin (Epo) hyporesponsiveness in patients with chronic kidney disease (CKD). Whether sFas could predict the need for erythropoiesis-stimulating agent (ESA) usage and its influence in erythropoiesis remain unclear. We evaluated the relation between sFas and ESA therapy in patients with CKD with anemia and its effect on erythropoiesis in vitro. First, we performed a retrospective cohort study with 77 anemic patients with nondialysis CKD. We performed in vitro experiments to investigate whether sFas could interfere with the behavior of hematopoietic stem cells (HSCs). HSCs were isolated from umbilical cord blood and incubated with recombinant sFas protein in a dose-dependent manner. Serum sFas positively correlated with Epo levels (r = 0.30, P = 0.001) but negatively with hemoglobin (r = -0.55, P < 0.001) and glomerular filtration rate (r = -0.58, P < 0.001) in patients with CKD at baseline. Elevated sFas serum levels (4,316 ± 897 vs. 2,776 ± 749, P < 0.001) with lower estimated glomerular filtration rate (26.2 ± 10.1 vs. 33.5 ± 14.3, P = 0.01) and reduced hemoglobin concentration (11.1 ± 0.9 vs. 12.5 ± 1.2, P < 0.001) were identified in patients who required ESA therapy compared with patients with non-ESA. Afterward, we detected that the sFas level was slight correlated with a necessity of ESA therapy in patients with nondialysis CKD and anemia. In vitro assays demonstrated that the erythroid progenitor cell frequency negatively correlated with sFas concentration (r = -0.72, P < 0.001). There was decreased erythroid colony formation in vitro when CD34+ HSCs were incubated with a higher concentration of sFas protein (1.56 ± 0.29, 4.33 ± 0.53, P < 0.001). Our findings suggest that sFas is a potential predictor for ESA therapy in patients with nondialysis CKD and that elevated sFas could affect erythropoiesis in vitro.