Cytomorphological features of forensic veterinary determination of the age of wounds inflicted by sharp instruments in dogs and cats

The urgency of the problem of postoperative wound healing after combined operations for combined pathology of the anal canal and rectum is quite high and contributes to the introduction into the practice of coloproctologists of new modern surgical technologies for the treatment of this pathology. The aim of the study was to conduct a comparative morphological assessment of postoperative wound healing in patients with combined pathology of the anal canal and rectum after combined operations using modern high-frequency electrosurgical and radiosurgical technologies. The results of surgical treatment of 689 patients with combined pathology of the anal canal and rectum using high-frequency electrosurgery and radiowave surgery with morphological assessment of wound healing on 3, 5, 7, 14, 21 days of the postoperative period, which were divided into 4 study groups. Using of “Surgitron” and “KLS Martin” devices for the treatment of patients with combined pathology of the anal canal and rectum was accompanied by the formation of the thinnest layers of coagulation necrosis in tissues with a depth of 0.189±0.085 mm and 0.194±0.090 mm respectively and as result patients of the first and fourth study groups had the shortest duration of inpatient treatment, which was 3-5 days and the average time of wound healing, which was 14-15 days. Patients in these study groups had the lowest inflammatory neutrophil reaction in postoperative wounds on day 3, which rapidly disappeared by day 5, on days 7-14 they had active reparative processes with the appearance of fibroblasts and connective tissue fibers, and on 21 day squamous epithelial cells, which indicated the processes of active epithelialization of wounds. The effect on the tissues of the devices “EFA” and “ERBE ICC 200” was deeper than in the above groups, forming a layer of coagulation tissue necrosis with a depth of 0.208±0.097 mm and 0.302±0.107 mm respectively, which was accompanied by patients of the third and second study groups with longer terms of inpatient treatment, which amounted to 5-7 days and increase the duration of wound healing, which amounted to 16-19 days. Patients in the 2nd and 3rd study groups showed a more pronounced inflammatory neutrophilic reaction in postoperative wounds on the 3rd day, which did not disappear until the 5th day and in half of the cases the presence of a significant number of segmental neutrophils and bacterial accumulations persisted. On days 7-14 they had weak reparative processes with the appearance of single fibroblasts and a small number of connective tissue fibers and on the 21st day single squamous epithelial cells, which indicated slow processes of wound epithelization. Using of radio-wave surgery and high-frequency electrosurgery devices promotes active epithelialization of tissues preventing scar strictures of the anal canal and improves the rehabilitation of patients in the postoperative period.

Differences in intestinal mucin dynamics between germ-free and conventionally reared chickens after mannan-oligosaccharide supplementation

This Month in Gastroenterology

Penicillium marneffei is an important opportunistic fungal pathogen. Host defence mechanisms against P. marneffei are not fully understood. We investigated the fungicidal activity of murine peritoneal macrophages against two forms of P. marneffei, conidia and yeast cells, and the involvement of the NO-mediated killing system. Peritoneal macrophages suppressed the intracellular growth of P. marneffei yeast cells and conidia. The number of live yeast cells within macrophages was significantly reduced by activation of macrophages by interferon-gamma (IFN-gamma), while a similar response was not observed with conidia. IFN-gamma-induced macrophage fungicidal activity against yeast cells was mediated by NO and was almost completely inhibited by N(G)-monomethyl-L-arginine (L-NMMA), a competitive inhibitor of NO synthesis, while N(G)-monomethyl-D-arginine (D-NMMA), an optical isomer of L-NMMA, did not show any influence. NO production by macrophages stimulated with IFN-gamma was significantly enhanced when these macrophages were cultured with P. marneffei yeast cells, while conidia did not enhance macrophage NO production. Furthermore, yeast cells were more susceptible to the killing effect of chemically generated NO than conidia. Our results indicate that the yeast form of P. marneffei is more sensitive to the fungicidal activity of IFN-gamma-stimulated macrophages than conidia, and suggest that the different effects of two forms of P. marneffei on macrophage NO production and their different susceptibilities to NO may be reasons for the present findings.

Erratum: High-Throughput Identification of Resistance to Pseudomonas syringae pv. Tomato in Tomato using Seedling Flood Assay.

Background: HAIs are infections that are obtained from hospitals or other health care facilities, such as health centers. According to the Ministry of Health, in Indonesia it reaches 15.74%, much higher than in developed countries, which ranges from 4.8 to 15.5%. According to the Indonesian Ministry of Health, the average is 8.1%. At the Puskesmas, the Emergency Unit (ER) is the place most at risk for HAIs, for patients undergoing treatment using less sterile equipment, such as minor surgical equipment. Object: To determine the presence or absence of bacteria and to identify the types of bacteria in minor surgical medical devices in the ER Pahandut Public Health Center, Palangka Raya City. Methods: This study used a descriptive quantitative design method, to determine the presence or absence of bacteria in sterilized minor surgical medical devices. Results: The identification results found gram-positive bacteria in 3 samples of minor surgical instruments, namely 2 samples identified as Staphylococcus aureus and 1 sample identified as Streptococcus sp. Conclusion : There are bacterial contaminants in minor surgical medical devices used in the ER Pahandut Public Health Center, Palangka Raya City.

Apoptosis and extracellular matrix–cell interactions in kidney disease

The development of graft-versus-host reactions in mice was characterized by an increase in activated macrophage populations in the peritoneum and spleen of the animals. In the present study we tested the effect of the immunosuppressive agent CsA on the appearance and activity of such macrophages. Parental spleen lymphocytes were injected intraperitoneally into F1 hybrids (BALB/c x C57Bl/6), and 13 days following injection we monitored the number of peritoneal exudate cells (PEC), spleen enlargement, and the oxidative burst (OB) of adherent peritoneal macrophages (APM). Macrophage OB was assessed by measuring O2- and H2O2 production, following stimulation with 12-O-tetradecanoyl-phorbol-13-acetate (TPA). In GVHR mice a 72% increase in their spleen to body weight ratio was observed, whereas in CsA-treated GVHR mice only a 32% increase was evident. Assessment of the effect of CsA on the number and function of peritoneal cells revealed that CsA caused a 73% reduction in the number of infiltrating PEC in GVHR mice. Furthermore, measurements of H2O2 and O2- production by APM revealed that the overall OB capacity of APM from CsA-treated mice was significantly reduced compared to APM from nontreated GVHR mice. CsA had no effect on the number and OB activity of paraffin oil elicited or resident PEC. These results indicate that CsA may prevent recall and activation of macrophages via its effect on T cell lymphokine release and thus may lessen the contribution of macrophage-derived toxic reagents to the damage inflicted by GVHR.

The effect of chemotherapy combined with immunostimulants on the activities of macrophages in mice was studied. The number of macrophages and exudate cells in the peritoneal cavity increased 3 days after ip injection with mitomycin-C, cyclophosphamide, and 5-fluorouracil together with OK-432 or yeast cell wall and decreased to normal level after 9 days, while the number of the cells remained decreased in mice receiving multi-drugs alone. Acid phosphatase activity of the macrophages of mice was elevated after the simultaneous injection of yeast cell wall and OK-432, and high activity was preserved in the macrophages of mice receiving yeast cell wall even after 9 days. Spreading of these cells was also enhanced. Macrophage activities examined by these assays were maximal in every respect 6 days after combination therapy. Cytostatic activity of the cells was strengthened after 6 days by combined use of OK-432 or yeast cell wall. Role of the activated macrophages in combination therapy was discussed.

The poor ability of injured central nervous system (CNS) axons to regenerate has been correlated, at least partially, with a limited and suppressed postinjury inflammatory response. A key cell type in the inflammatory process is the macrophage, which can respond in various ways, depending on the conditions of stimulation. The aim of this study is to compare the activities of macrophages or microglia when encountering CNS and peripheral nervous systems (PNS), on the assumption that nerve-related differences in the inflammatory response may have implications for tissue repair and thus for nerve regeneration. Phagocytic activity of macrophages or of isolated brain-derived microglia was enhanced upon their exposure to sciatic (PNS) nerve segments, but inhibited by exposure to optic (CNS) nerve segments. Similarly, nitric oxide production by macrophages or microglia was induced by sciatic nerve segments but not by optic nerve segments. The previously demonstrated presence of a resident inhibitory activity in CNS nerve, could account, at least in part, for the inhibited phagocytic activity of blood-borne macrophages in CNS nerve as well as of microglia resident in the brain. It seems that the CNS microglia are reversibly immunosuppressed by the CNS environment, at least with respect to the activities examined here. It also appears from this study that the weak induction of early healing-related activities of macrophages/microglia in the environment of CNS might explain the subsequent failure of this environment to acquire growth-supportive properties in temporal and spatial synchrony with the needs of regrowing axons.

Endothelin-1 increases expression and activity of arginase 2 via ETB receptors and is co-expressed with arginase 2 in human atherosclerotic plaques

We studied reparative effect of platelet-filled biological matrixes in the treatment of mice with wounds equivalent to deep burn. The wound coatings were based on decellularized dermal matrix without platelets (control), with native platelets, and with platelets stabilized with 2.5 μM nanosilver. In 3 days, the epithelial layer and derma were absent in all groups and extensive scab was formed. Dermal matrix with platelets simulated intensive migration of macrophages and fibroblasts to the wound bottom; in the control group, this migration was absent. In 14 days, granulation tissue appeared in the wound bottom in animals of all groups; in the experimental groups, the number of vessels was 2-4-fold higher than in the control, though the number of inflammatory cells in experimental groups remained high. On day 21, the scab on the most of the wound area was absent in all animals of the experimental groups and epithelialization and hair growth were pronounced, comparing to control. Nevertheless, in experiment dermal layer was not already completed, inflammation reaction remained.

BACKGROUND: Purulent diseases of the skin and subcutaneous tissue remain a widespread and urgent problem. The most promising methods of treatment with the active use of physical factors affecting the purulent focus are in the conditions of modern increasing resistance of microorganisms. AIM: To determine the effect of local ultra-low temperatures on the inflammatory and reparative processes in an experimental septic wound of soft tissues for planning a subsequent clinical study of the effectiveness of debridement using cryotherapy. MATERIALS AND METHODS: The experiment was conducted on 40 Wistar–Kyoto male rats of 9-month age, weighing 200 to 230 grams. The wound was created by excising an oval skin flap 2.0 × 1.5 cm in the withers region, followed by injection of 1.5 ml of Staphylococcus aureus solution at a concentration of 109 CFU/1 ml into the hypodermis. The animals were divided to 4 groups: a control group with the natural course of the healing process, and the main groups, which included a group with single daily treatment of the wound with Levomekol ointment, a group with a single daily treatment of the wound with vapor-drop jet of liquid nitrogen, and a group with single daily treatment with a combination of the above methods. The treatment was carried out during the exudative phase of the wound process. With the disappearance of exudation and of paravulnar edema and hyperemia, debridement in the main groups was stopped. In the dynamics, a comparative observation and study of planimetric changes in the wound were carried out. RESULTS: A more rapid decline of the severity of hyperemia, paravulnar edema and exudation, accelerated filling the wound with granulation tissue, accelerated epithelialization were seen in treatment with cryotherapy compared with treatment with Levomekol. These effects led to increase in the rate of wound healing due to the shortening of all phases of the wound process by at least 2 days. There were no statistically significant side effects, healing and survival occurred in all animals. CONCLUSION: In was found in the study that the use of cryotherapy leads to a rapid subsidence of symptoms, activates the processes of granulation and epithelization. Mutual potentiation of the positive effects of debridement using ultra-low temperatures and antibacterial agents has been established. Cryotherapy proved to be a safe and effective means of treating septic wounds, which does not prevent and makes it possible to conduct further clinical studies.

In this regard, the cytological picture of the lymphoid tissue in calves’ inguinal lymph node with spontaneous leptospirosis was studied. The study material was taken from the inguinal lymph node of the 11 calves who died of leptospirosis during the enzootic period in Azerbaijan. The study material samples were fixed in 10% neutral formalin solution, followed by pouring in paraffin, coloring azur sections with 2-eosin and counting 13 cell types (lymphoblast, prolymphocyte, lymphocyte, free reticular cell, process sinus reticular cell, endothelium, fibroblast, histiocyte, macrophage, polyblast, plasmablast, protoplasmocyte, plasmacyte) using MOV-15. It was established that the number of lymphoblasts in the inguinal lymph node with subacute leptospirosis decreased 2.2 times, the number of prolymphocytes decreased 1.4 times, the number of lymphocytes decreased 4.4 times. The number of free reticular cells from the cells of the reticuloendothelium decreased 3.7 times. However, the number of grown sinus reticular cells and the endothelium of the sinuses fluctuated within the normal range. The number of fibroblasts increased 1.7 times, histiocytes - 6.6 times, macrophages - 11.8 times, and polyblasts - 11 times (Table 1). At the same time, there was a sharp increase in the number of cells in the plasma row. Of those, the number of plasmablasts increased 8.5 times, protoplasmocytes - 30.4 times, plasma cells - 17 times. Overall, the cytological picture in the inguinal lymph node during spontaneous leptospirosis in calves was characterized by an increase in the number of plasma cells, fibroblasts, histiocytes, macrophages, polyblasts and a decrease in the number of lymphoblasts, prolymphocytes, lymphocytes and free reticular cells.

The p38 mitogen-activated protein kinase (MAPK) pathway, like the c-Jun N-terminal kinase (JNK) MAPK pathway, is activated in response to cellular stress and inflammation and is involved in many fundamental biological processes. To study the role of the p38 MAPK pathway in vivo, we have used homologous recombination in mice to inactivate the Mkk3 gene, one of the two specific MAPK kinases (MAPKKs) that activate p38 MAPK. Mkk3(-/-) mice were viable and fertile; however, they were defective in interleukin-12 (IL-12) production by macrophages and dendritic cells. Interferon-gamma production following immunization with protein antigens and in vitro differentiation of naive T cells is greatly reduced, suggesting an impaired type I cytokine immune response. The effect of the p38 MAPK pathway on IL-12 expression is at least partly transcriptional, since inhibition of this pathway blocks IL-12 p40 promoter activity in macrophage cell lines and IL-12 p40 mRNA is reduced in MKK3-deficient mice. We conclude that the p38 MAP kinase, activated through MKK3, is required for the production of inflammatory cytokines by both antigen-presenting cells and CD4(+) T cells.