Irina Chekarova

Importance of pericytes and mechanisms of pericyte loss during diabetic retinopathy

Pericytes are distinctive regulators of angiogenesis and are adumbrated to provide vessel stability and control of endothelial proliferation. The present article spotlights the persona of pericytes in physiological angiogenesis, recruitment of pericytes and different mechanisms of pericyte depletion. Developing retina appears particularly dependent on pericytes, and pericyte loss is considered as hallmark of early diabetic retinopathy. Several factors are contemplated to be engaged in pericyte conscription including angiopoietin‐1 and its receptor tyrosine kinase Tie‐2, vascular endothelial growth factor‐A and its receptor flk‐1 and the platelet‐derived growth factor PDGF‐B/PDGF‐β system. At present, the mechanisms by which diabetes persuade apoptosis in the retinal microvasculature remain indecisive, albeit oxidative stress, formation of advanced glycation end products , upregulation of protein kinase C, increased polyol pathway flux and focal leukostasis may be important. In this context, accelerated microvascular cell death may become a constructive surrogate end‐point in pharmacological studies of experimental diabetes.

Effect of aged garlic extract on wound healing: a new frontier in wound management.

Successful wound healing depends upon angiogenesis, and impaired angiogenesis is a hallmark of the chronic wounds encountered with diabetes and venous or arterial insufficiency. To intervene and improve wound closure, it is essential to investigate the effects of different natural remedies in wound healing. The chicken dorsum skin excisional wound assay was used to investigate the influence of different concentrations of aged garlic solution (AGS) on wound healing. Gross, histopathology, scanning electron microscopy (SEM) and computer-based three-dimensional (3D) image-probing techniques were utilized to determine the effects of AGS on wound closure, re-epithelialization, dermal matrix regeneration, and angiogenesis. Ninety chicks, aged 1 week and divided in 6 groups, were topically exposed to different concentrations of AGS for 6 days: control (group A), 1% (group B), 5% (group C), 10% (group D), 15% (group E), and skin lotion (group F). Different patterns, ranging from incomplete to almost complete wound closure, were observed among different groups with highly significant results (P < 0.001) in group E. Histological investigations revealed a positive augment in the re-epithelialization of all AGS exposed wounds. An increase in the number of new loosely packed collagen and maturation of collagen bundles was observed in all treated wounds at days 4 and 6 post-wounding, respectively. Similar results were achieved through SEM of treated wounds. Histological investigations revealed the profuse dose-dependent neovascularization among AGS-treated wounds. Abbott curve, angular spectrum, and different parameters of 3D surface roughness of wounds were also measured for the precise quantification of angiogenesis. A very highly significant (P < 0.001) increase in angiogenesis was observed among all treated groups. No significant change was observed among control and skin lotion–treated groups. These observations substantiate the beneficial use of AGS in the treatment of wounds. Additional studies are needed to study the specific wound-healing mechanisms of chemical, or group of chemicals, present in AGS.

A Novel 3-D Model of Chick Chorioallantoic Membrane for Ameliorated Studies in Angiogenesis

Decisive indulgence of angiogenesis requires a more holistic assessment and several in vivo assays have been developed that permit a more realistic appraisal of the angiogenic response. One of the most popular assays to study angiogenic activity is the chick chorioallantoic membrane (CAM). Although CAM assay is a vital technique used to study normal and putative angiogenesis, a serious drawback in its utilization is the lack of quantitative assessment of vascularization. In this study, we proposed a new 3-D model of the developing CAM for precise quantification of normal vasculature of CAM from Day 4 to Day 13 of incubation. Image probing technique was used to quantify different 3-D parameters of vascular microarchitecture. A significant increase (P < 0.05) in surface roughness (Sa) was observed at Day 5 of incubation, while highly significant increase (P < 0.01) in Sa values was observed at Day 6 of incubation. Maximum increase (P < 0.001) in Sa values was observed from Day 7 to Day 9 of incubation and slight decrease in Sa values was observed in successive days. Similar results were observed for root mean square values (Sq), absolute heights of the surface (Sz), and amount of lowest valleys (Sy) on CAM. A significant increase (P < 0.05) in developed surface area (Sdr) from Day 5 of incubation, reaching to its maximum (P < 0.01) at Day 9 also was noted. Similarly, significant increase (P < 0.05) in fluid core retention (Sci) illustrates presence of copious fluid in blood vessels. Image probing technique offers a useful modality for visualizing 3-D microvascular architecture of CAM to exaggerate the fine details and reveal the hidden information that can be helpful for precise quantification of angiogenesis. This approach can be used to evaluate the angiogenic and antiangiogenic potential of different biological substances and also can be a valuable independent prognostic indicator in a wide variety of human cancers.

Pollution dilemma in Asian population: CNG and wound healing.

Automobile exhaust constituents contribute significantly to air pollution in urban areas and compressed natural gas (CNG) is considered one of the most promising fuel alternatives for the future. CNG-powered four-stroke engine auto-rickshaws are ubiquitous in South Asian cities as taxi and for commercial transportation. Automotive exhaust contains several toxins, which are overwhelmingly toxic to the processes of wound healing. By utilizing the in vivo mouse model of wound healing, this report analyzes the effects of CNG-powered four-stroke auto-rickshaws smoke solution (4SARSS) on different events of wound healing; dermal matrix regeneration, re-epithelialization and neovascularization. A total of 72 adult mice, divided in eight groups were exposed to 4SARSS for 12 days. A highly significant reduction (P<0.001) in wound closure was observed among all 4SARSS treated groups, at each time point of the experiment. An immature development in both the neoepidermis and the neodermis was observed among all 4SARSS treated wounds with defective re-epithelialization, dermal matrix regeneration and maturation of collagen bundles. Abbott curve, angular spectrum, 3D surface topographies, and histological investigations of wounds explicated highly significant activation (P<0.001) of delayed-neovascularization among 4SARSS treated wounds. All these annotations advocate excessive toxicity of emission from CNG-powered auto-rickshaws to the process of wound healing and people occupationally exposed to this toxic emissions may suffer varying degree of delayed wound healing.

Granulomatous pneumonia in a captive freshwater crocodile (Crocodylus johnstoni) caused by Mycobacterium szulgai.

Abstract A 25-yr-old male freshwater crocodile (Crocodylus johnstoni) was diagnosed with pulmonary mycobacteriosis caused by Mycobacterium szulgai. Necropsy revealed fibrinous exudate in the right pleural cavity and white miliary nodules in the right lung lobe. Histopathologic examination revealed well-demarcated granulomas consisting of multinucleated giant cells and epithelioid cells surrounded by fibrous connective tissue. Atypically, lymphocytes had accumulated in the outer region of fibrous connective tissue. Mycobacterial infection was confirmed by nested polymerase chain reaction targeting the hsp65 gene and by Fites method for detection of acid-fast bacilli within formalin-fixed, paraffin-embedded lung tissue. Sequence analysis of the DNA amplicon revealed that the species of mycobacterium shared 98% homology with the gene encoding the hsp65 gene of M. szulgai. This is the first report of M. szulgai as the causative agent of mycobacteriosis in a reptile.

Cytotoxicity of Fumonisin B1 in Spheroid and Monolayer Cultures of Rat Hepatocytes

Fumonisin B1 (FB1), the most prevalent member of toxins produced by several species of Fusarium molds, which occur mainly in maize, causes several fatal hepatopathies and nephropathies of animals. The current study was scrutinized to ascertain different cytotoxic and morphological transformations in rat hepatocytes induced by the treatments of diverse concentrations (300, 500, or 1000 μM) of fumonisin B1 in vitro, using both monolayer and spheroid cultures. In each hepatocyte culture, the cytotoxicity of FB1 was augmented in dose- and time-response manners. Morphological transformations among FB1-treated groups integrated accumulation of lipid droplets, cytoplasmic vacuolation in hepatocyte monolayers, and bleb formation in the hepatocyte spheroids. Additionally, electron microscopy revealed the loss of microvilli, mitochondrial swelling, and formation of lamellar membranous whorl in the vacuoles and bile canaliculi-like structures. Appearance of electron dense bodies in the monolayers, and loss of cell-to-cell contact in spheroids were depicted in 1000 μM FB1-treated hepatocytes. These outcomes insinuate different vital events in explaining morphological transformations in the cell membrane and organelles, induced by fumonisins in rat hepatocytes.