Exosomal miRNA-455 from Bone Marrow Stromal Cells (BMSCs) Promotes Macrophage Phagocytosis and Restrains Progression of Gastric Cancer (GC)

Multiple miRNAs are differentially expressed in gastric cancer (GC). Herein, this study aims to investigate miR-455’s role in GC and its mechanism. Exosomes (exo) separated from BMSCs after transfection were co-cultured with either phagocytes, GC cells (NCI-N87 cell), or macrophages combined with NCI-N87cells (mixed group) followed by analysis of the expression of PTEN, N-cadherin, E-cadherin, and PI3K, and AKT by RT-qPCR and Western blot. Increased miR-455 expression was observed in GC cells upon transfection. GC cells in the mixed group relative to NCI-N87 group exhibited a lower cell migration and invasion and impaired proliferative capacity (p < 0.05), accompanied with higher expressions of N-cadherin, E-cadherin, PI3K, and AKT, and decreased level of PTEN (p < 0.05). The combined treatment resulted in a higher phagocytic rate (12.38±0.21%) and phagocytic index (14.29±2.11%) compared to treatment with only phagocytes (p < 0.05). In conclusion, BMSC-derived exosomal miR-455 inhibits the growth of GC cells and promotes the phagocytosis through inactivating PI3K/AKT signaling pathway.

Veterinary drug Trametin obtained on the basis of Trametes pubescens xylotroph fungi: its effect on the biosynthesis of interferons and its prophylactic activity against calf respiratory diseases

Given the spread of bacterial and viral diseases in young farm animals, the use of interferons and drugs to stimulate their biosynthesis has gained relevance. In a previous study, we examined the effect of a veterinary drug Trametin produced on the basis of Trametes pubescens (Shumach.: Fr.) Pilat. on the biosynthesis of interferons in the blood of mice. The present work is aimed at studying the biosynthesis dynamics of α- and γ-interferons when using Trametin and studying its prophylactic activity in calves. It is shown that a single oral administration of Trametin in doses ranging from 15 to 60 mg/kg causes a dose-dependent induction and production of γ-interferon in the blood of mice, whose maximum content reaches 1337.0±93.0 pg/mL at 48 h after administering a dose of 30 mg/kg. With a Trametin dose increase from 15 to 30 mg/kg, the level of α-interferon production rises to 1388.0±84.0 pg/mL at 48 h after administration. At a Cycloferon dose of 4.5 mg/kg, the production level of α-interferon and γ-interferon amounts to 1455.47±84.2 and 1447.0±90.0 pg/mL, respectively. The immunostimulatory properties of Trametin are confirmed by a scientific and economic experiment conducted using immunocompromised calves. In our studies, an immunological test of calf blood performed prior to and following the administration of Trimetin and Cycloferon constitutes criteria for the prophylactic activity of these drugs. The prophylactic efficacy of Trametin is confirmed by an increase in phagocytic activity by 10.5%, phagocytic index by 61.8%, and phagocytic number by 52.8%. After Trametin administration, the bactericidal activity of the serum increases by 60%. Cycloferon exhibits a similar immunostimulatory effect. Nonspecific prophylaxis using Trametin is shown to reduce the incidence of bacterial and viral respiratory diseases in young calves and generally improve their immunity.

Immunostimulant Activity of Marchantia paleacea Bertol. Herb Liverwort Ethanol Extract in BALB/c Mice

Immunostimulants are compounds that can stimulate an immune response by increasing the activity of non-specific and specific components of the immune system (humoral and cellular) against certain infections and diseases. The liverwort plant species Marchantia paleacea Bertol. has long been used as a source of nutrition and empirical medicine. However, scientifically there is still not much research data on immunomodulators in these plants. This study aims to determine the activity of immunomodulators in the ethanol extract of the herb Marchantia paleacea Bertol. in male mice of BALB/c strain. Bioactive compounds from this plant were extracted by maceration method using 96% ethanol. Extract characterization and phytochemical screening were determined according to WHO guidelines and standard procedures from previous studies. The immunomodulatory activity of the extract was tested by carbon clearance method and lymphoid organ index (non-specific responses), primary and secondary antibody titer tests (humoral specific responses), IL-2 cytokine levels and IFN-ɣ from serum secondary antibodies and delayed-type hypersensitivity reaction/DTH (cellular specific response). The results of qualitative phytochemical screening contained flavonoid compounds, saponins, phenolics, tannins and steroids/triterpenoids. The results of the non-specific immune response immunomodulator test showed that the dose of 52 mg/kg bw had the largest phagocytic index of 1.52 which included strong immunostimulation (K > 1.5) and the organ spleen index of 0.55 ± 0.11 which increased significantly compared to the control (p<0.05). The data on the acquisition of specific immune responses in the primary and secondary antibody titer test in the three test extracts resulted in increased titer levels compared to the control and at a dose of 52 mg/kg bw could significantly increase the levels of IL-2 cytokines in the control group (p<0,05). Meanwhile, in the DTH test, doses of 13 and 26 mg/kg bw could significantly increase the thickness of the soles of mice compared to controls (p<0.05).

Phagocytic and oxidative burst activity of neutrophils in type 2 diabetic patients with foot ulcers

Introduction and Aim: Diabetic foot ulcers are common complications seen in diabetic patients. Treatment of this disabling foot sore remains a challenge to health care professionals. This study aimed at evaluating whether the neutrophils from type 2 diabetic patients with foot ulcers present an impairment of phagocytic index and impairment in respiratory burst. We also aimed at understanding whether the impairment in neutrophil phagocytic activity can be alleviated with short course of standard treatment regime for foot ulcers. Methodology: For this case-controled study, 43 participants with type 2 diabetes (18 with foot ulcers and 25 without foot ulcers) were prospectively recruited along with 18 healthy volunteers. Phagocytic activity of neutrophils and respiratory burst of neutrophils was assessed along with ESR, percentage neutrophil counts before and after 2 weeks of standard treatment for foot ulcers. Results: Neutrophils of type 2 diabetic patients (with and without foot ulcers ) showed lower levels of phagocytic index and lower percentage of respiratory burst on comparison with non-diabetic subjects. Furthermore, on receiving treatment for foot ulcers, a significant improvement in neutrophil phagocytic indices were observed, along with improvement in wound ulcer score. Conclusion: Phagocytic activity of the neutrophils is impaired in type 2 diabetics (with and without foot ulcers). Neutrophil phagocytic indices can be improved on glycemic control. Additionally, improvement in neutrophil phagocytic indices after short course treatment for foot ulcers can be useful markers to predict treatment efficacy and in prognosis of diabetic foot ulcers.

Laboratory evaluation of the efficacy of Liastene application in complex therapy of multiresistant pulmonary tuberculosis

OBJECTIVE. Evaluating according to laboratory tests the effectiveness of Liasten in the treatment of patients with multidrug-resistant pulmonary tuberculosis (MDR-TB). MATERIALS AND METHODS. Evaluation of the effectiveness of etiotropic and etiopathogenetic therapy in 57 patients with MDR-TB was performed. According to the treatment schemes, patients were divided into groups. The control group (n=22) received individualized antimycobacterial therapy (AMBT) regimens. The experimental group (n=35) received AMBT in combination with Liasten. Evaluation of the effectiveness of treatment regimens was performed on the basis of indicators of general clinical blood tests, immunological and bacteriological studies. RESULTS AND DISCUSSION. In patients of the experimental group, compared with the control in 1.5 times more often found positive changes in the hemogram of blood and ESR (p<0.05-0.001), the establishment of a dynamic balance between the pools of lymphocyte cells CD4+ and СD8+ (immunoregulatory index, p<0.05), an increase in the number of phagocytosis active cells (phagocytic index, p<0.05), the content of cationic lysosomal proteins of granulocyte leukocytes (p<0.05), a 1.4-fold decrease in the cytochemical coefficient of neutrophils (p<0.05), the number of proliferated under the action of PPD-L lymphocytes (p<0.05), normalization of phagocytic counts and total redox activity of neutrophils (p<0.05), increase in frequency and reduction of anesthesia was stated. CONCLUSIONS. Restoration of the body’s immune status, blood hemogram, increase in frequency and reduction of the time of decontamination were more active and occurred 1.5 times more often in patients receiving a complex combination of AMBT with Liasten.

Nonspecific protective factors in patients with early latent syphilis (report 1)

The objective of this work is to study nonspecific protective factors in patients with latent early syphilis. The results of the study are based on the data of a comprehensive examination before treatment of 142 patients with early latent syphilis and 20 patients of the comparison group. Methods of investigation: serological methods for the diagnosis of syphilis complex of serological reactions, enzyme-linked immunosorbent assay (IgM, IgG), Indirect immunofluorescence reaction (RIF) RIF-200, RIF-abs; determination of cytokines IL-2, IL-6, IL-10, TNFα and INFγ in blood serum by enzyme-linked immunosorbent assay; determination of the phenotype of lymphocytes (CD-receptors), the concentration of circulating immune complexes, phagocytic number, phagocytic index, Nitroblue tetrazolium test. In the blood serum of patients the concentration of IL-2, IL-6, IL-10 and TNFα, INFγ was significantly increased. The concentration of IL-10 and IL-6 was also significantly increased in patients with more than 1 year of infection. The most significant disorders of the parameters of the metabolic activity of neutrophils were found in patients with an infection of more than 1 year; an imbalance in the circulating immune complexes concentration was also established. Under the increasing duration of the infection, the concentration of CD16+ in the blood serum progressively decreases. The content of pro-inflammatory and anti-inflammatory cytokines in the blood serum of patients with early latent syphilis was significantly increased and depended on the timing of infection. This relationship was most pronounced for IL-6 and IL-10. In patients with early latent syphilis there is also an imbalance between CD25+ and late activation factor HLA-DR against the background of a progressive decrease in the number of CD16+ lymphocytes. A direct relationship was established between the time of infection and the number of NK-cells. The revealed disorders may be the consequence of significant disorders on the part of neutrophils and may be one of the factors of the latent course of syphilitic infection.

Long-term immunity alterations in the employees of the high hydrogen sulfide content gas condensate processing facility

The current measures for protection of the gas processing plant employees cannot fully prevent the impact of pollutants. Evaluation of the immune system is one of the methods for monitoring of the employees' health, and testing the system of measures used to improve the working conditions. The study was aimed to identify alterations in the immune status of the employees at the gas processing and high hydrogen sulfide content condensate processing facility depending on their working experience. The working environment and the employees' immune system were evaluated by standard methods. Pollutants were detected with the Bruel & Kjaer 1302 Multi-Gas Monitor, and the Tsvet-550 gas chromatographer. A total of 160 employees and 81 controls (blood donors of the regional blood transfusion station) were surveyed. The immune system was evaluated using the System 9000 Plus hematological analyser, Cyto FLEX LX flow cytometer, UNICO 2100UV specrophotometer, and KFK-3-03-ZОМЗ photometer. It was concluded that the existing complex of occupational and industrial hazards affects the immune status of the main production unit employees, which is reflected in the decreased CD20 levels and increased CD8 levels along with the constant levels of CD4. Correlations were revealed between the immuniglobuline level alterations, decrease in the phagocytic index and phagocytic number, as well in lysozyme activity, and the working experience. Pollutant exposure results in altered immunity of the employees, which could be considered the adaptation mechanism.

SARS-CoV-2 and the Immune Response in Pregnancy with Delta Variant Considerations

As of September 2021, there has been a total of 123,633 confirmed cases of pregnant women with SARS-CoV-2 infection in the US according to the CDC, with maternal death being 2.85 times more likely, pre-eclampsia 1.33 times more likely, preterm birth 1.47 times more likely, still birth 2.84 times more likely, and NICU admission 4.89 times more likely when compared to pregnant women without COVID-19 infection. In our literature review, we have identified eight key changes in the immunological functioning of the pregnant body that may predispose the pregnant patient to both a greater susceptibility to SARS-CoV-2, as well as a more severe disease course. Factors that may impede immune clearance of SARS-CoV-2 include decreased levels of natural killer (NK) cells, Th1 CD4+ T cells, plasmacytoid dendritic cells (pDC), a decreased phagocytic index of neutrophil granulocytes and monocytes, as well as the immunomodulatory properties of progesterone, which is elevated in pregnancy. Factors that may exacerbate SARS-CoV-2 morbidity through hyperinflammatory states include increases in the complement system, which are linked to greater lung injury, as well as increases in TLR-1 and TLR-7, which are known to bind to the virus, leading to increased proinflammatory cytokines such as IL-6 and TNF-α, which are already elevated in normal pregnant physiology. Other considerations include an increase in angiotensin converting enzyme 2 (ACE2) in the maternal circulation, leading to increased viral binding on the host cell, as well as increased IL-6 and decreased regulatory T cells in pre-eclampsia. We also focus on how the Delta variant has had a concerning impact on SARS-CoV-2 cases in pregnancy, with an increased case volume and proportion of ICU admissions among the infected expecting mothers. We propose that the effects of the Delta variant are due to a combination of (1) the Delta variant itself being more transmissible, contagious, and efficient at infecting host cells, (2) initial evidence pointing to the Delta variant causing a significantly greater viral load that accumulates more rapidly in the respiratory system, (3) the pregnancy state being more susceptible to SARS-CoV-2 infection, as discussed in-depth, and (4) the lower rates of vaccination in pregnant women compared to the general population. In the face of continually evolving strains and the relatively low awareness of COVID-19 vaccination for pregnant women, it is imperative that we continue to push for global vaccine equity.

Growth Performance, Antioxidant Activity, Immune Status, Meat Quality, Liver Fat Content, and Liver Histomorphology of Broiler Chickens Fed Rice Bran Oil

This trial was performed to determine the effect of rice bran oil (RBO) inclusion in diets of broiler chickens on performance, carcass characteristics, blood parameters, meat quality, antioxidant activity, liver lipid content, and liver histological structure. The 35-day feeding trial was conducted on 240 one-day-old Ross 308 broiler chickens, allocated to four treatment groups with six replicates each. RBO was examined at different inclusion levels, 0% (control), 1% (RBO1%), 1.5% (RBO1.5%), and 2% (RBO2%) in a completely randomized design. The results showed that at the end of the trial (35 days) the RBO supplementation had positive effects (p < 0.001) on the productivity parameters, but the feed intake was linearly decreased due to RBO inclusion. In addition, RBO supplementation linearly improved (p < 0.05) the dressing percentage, breast yield, immune organs relative weights, and meat glutathione concentration, while it decreased (p < 0.01) the abdominal fat yield and meat crude fat, triglycerides, cholesterol, and Malondialdehyde (MDA) contents in broiler’s meat. Moreover, serum total protein, globulin, and high-density lipoprotein contents improved noticeably (p < 0.01) due to offering an RBO-supplemented diet, but serum total lipids, total cholesterol, triglyceride, low-density lipoprotein, and aspartate aminotransferase concentrations linearly reduced (p < 0.01). The RBO supplementation augmented (p < 0.05) the phagocytic index, phagocytic activity, and antibody titer compared to control. On the other hand, RBO inclusion had no effect on the breast, thigh, or abdominal fat color parameters. Moreover, RBO supplementation reduced (p < 0.01) the content of total saturated FA (SFA), but increased (p < 0.01) the content of total monounsaturated FA (MUFA), and polyunsaturated FA in both breast and thigh meat. Chemical analysis of the liver tissue samples revealed that the inclusion of RBO linearly reduced (p < 0.05) hepatic cholesterol, triglyceride, and MDA contents. Histologically, the lipid percentage and number of lipid droplets (p < 0.01) were markedly lessened in the RBO-supplemented groups. The histological structure of the liver asses by light and electron microscope were normal in all groups without any pathological lesions. It is concluded that RBO could be used as a valuable ingredient in broiler chickens’ diets to stimulate the growing performance and immune status, enhance the antioxidant activity and blood lipid profile, augment liver function, and improve the nutritive value of the meat.

The Role of Dimorphism Regulating Histidine Kinase (Drk1) in the Pathogenic Fungus Paracoccidioides brasiliensis Cell Wall

Dimorphic fungi of the Paracoccidioides genus are the causative agents of paracoccidioidomycosis (PCM), an endemic disease in Latin America with a high incidence in Brazil. This pathogen presents as infective mycelium at 25 °C in the soil, reverting to its pathogenic form when inhaled by the mammalian host (37 °C). Among these dimorphic fungal species, dimorphism regulating histidine kinase (Drk1) plays an essential role in the morphological transition. These kinases are present in bacteria and fungi but absent in mammalian cells and are important virulence and cellular survival regulators. Hence, the purpose of this study was to investigate the role of PbDrk1 in the cell wall modulation of P. brasiliensis. We observed that PbDrk1 participates in fungal resistance to different cell wall-disturbing agents by reducing viability after treatment with iDrk1. To verify the role of PbDRK1 in cell wall morphogenesis, qPCR results showed that samples previously exposed to iDrk1 presented higher expression levels of several genes related to cell wall modulation. One of them was FKS1, a β-glucan synthase that showed a 3.6-fold increase. Furthermore, confocal microscopy analysis and flow cytometry showed higher β-glucan exposure on the cell surface of P. brasiliensis after incubation with iDrk1. Accordingly, through phagocytosis assays, a significantly higher phagocytic index was observed in yeasts treated with iDrk1 than the control group, demonstrating the role of PbDrk1 in cell wall modulation, which then becomes a relevant target to be investigated. In parallel, the immune response profile showed increased levels of proinflammatory cytokines. Finally, our data strongly suggest that PbDrk1 modulates cell wall component expression, among which we can identify β-glucan. Understanding this signalling pathway may be of great value for identifying targets of antifungal molecular activity since HKs are not present in mammals.