MicroRNA-30a-5p promotes differentiation in neonatal mouse spermatogonial stem cells (SSCs)

Background The importance of spermatogonial stem cells (SSCs) in spermatogenesis is crucial and intrinsic factors and extrinsic signals mediate fate decisions of SSCs. Among endogenous regulators, microRNAs (miRNAs) play critical role in spermatogenesis. However, the mechanisms which individual miRNAs regulate self- renewal and differentiation of SSCs are unknown. The aim of this study was to investigate effects of miRNA-30a-5p inhibitor on fate determinations of SSCs. Methods SSCs were isolated from testes of neonate mice (3–6 days old) and their purities were performed by flow cytometry with ID4 and Thy1 markers. Cultured cells were transfected with miRNA- 30a-5p inhibitor. Evaluation of the proliferation (GFRA1, PLZF and ID4) and differentiation (C-Kit & STRA8) markers of SSCs were accomplished by immunocytochemistry and western blot 48 h after transfection. Results Based on the results of flow cytometry with ID4 and Thy1 markers, percentage of purity of SSCs was about 84.3 and 97.4 % respectively. It was found that expression of differentiation markers after transfection was significantly higher in miRNA-30a- 5p inhibitor group compared to other groups. The results of proliferation markers evaluation also showed decrease of GFRA1, PLZF and ID4 protein in SSCs transfected with miRNA-30a-5p inhibitor compared to the other groups. Conclusions It can be concluded that inhibition of miRNA-30a-5p by overexpression of differentiation markers promotes differentiation of Spermatogonial Stem Cells.

BATTERY OF TESTS FOR STUDYING THE ADAPTOGENIC EFFECTS OF BIOLOGICALLY ACTIVE SUBSTANCES IN PRE-CLINICAL TRIALS

The purpose of the article is to propose to researchers of the problems of adaptation and the study of specific adaptogenic activity standardized approaches to biomedical (preclinical) research of new biologically active substances, ensuring their comparability. The analysis of modern views on adaptation processes, as an increase in nonspecific resistance to adaptive effects, and the properties of known adaptogens is carried out. It has been shown that adaptogenic action can be realized through the involvement of several regulatory pathways, such as changes in the activity of neurons and endocrine response in response to adverse effects, interaction with cell receptors and modulation of their sensitivity to endogenous regulators, changes in the composition of cell membranes, structure of the cytoskeleton, activities of enzyme complexes, epigenomic regulation, antioxidant and antiradical activity. A grouping of methods for the experimental study of adaptogens is proposed on the basis of their compliance with the definition of this pharmacological class and the requirements of evidence-based medicine. The groups of methods aimed at increasing the rate and stability of the formation of a state of increased resistance to adverse influences, reparative-restorative processes, nonspecific resistance, psychoactivating and neuroplastic effects, antimutagenic and anticarcinogenic effects were identified. Specific schemes of adaptive influences and criteria for assessing adaptogenic activity are proposed on the basis of comparing the results obtained with the effects of "reference" adaptogens. The technology of the integrated assessment of the results of the methods, which are different in their informational significance, has been substantiated.

The Role of miR-107 in Prostate Cancer: A Review and Experimental Evidence

Over the past two decades, several research groups have focused on the functioning of microRNAs (miRNAs), because many of them function as positive or negative endogenous regulators of processes that alter during the development of cancer. Prostate cancer is the second most commonly occurring cancer in men. New biomarkers are needed to support the diagnosis of prostate cancer. Although it is necessary to deepen the research on this molecule to explore its potential utility in the diagnosis, follow-up, and prognosis of cancer, our results support a role of miR-107 in the signaling cascades that allow cancer progression, and as shown here, in the progression of Prostate Cancer (PCa). These findings strongly suggest that miR-107 may be a potential circulating biomarker for the diagnosis and prognosis of prostate cancer.

In situ characterisation of phytohormones from wounded Arabidopsis leaves using desorption electrospray ionisation mass spectrometry imaging

Phytohormones (plant hormones) are a group of small signalling molecules that act as important endogenous regulators in the plant development and stress responses. Previous research has identified phytohormone species, jasmonates,...

Resolution-Associated Molecular Patterns (RAMPs) as Endogenous Regulators of Glia Functions in Neuroinflammatory Disease

: Glial cells, including microglia and astrocytes, facilitate the survival and health of all cells within the Central Nervous System (CNS) by secreting a range of growth factors and contributing to tissue and synaptic remodeling. Microglia and astrocytes can also secrete cytotoxins in response to specific stimuli, such as exogenous Pathogen-Associated Molecular Patterns (PAMPs), or endogenous Damage-Associated Molecular Patterns (DAMPs). Excessive cytotoxic secretions can induce the death of neurons and contribute to the progression of neurodegenerative disorders, such as Alzheimer’s disease (AD). The transition between various activation states of glia, which include beneficial and detrimental modes, is regulated by endogenous molecules that include DAMPs, cytokines, neurotransmitters, and bioactive lipids, as well as a diverse group of mediators sometimes collectively referred to as Resolution-Associated Molecular Patterns (RAMPs). RAMPs are released by damaged or dying CNS cells into the extracellular space where they can induce signals in autocrine and paracrine fashions by interacting with glial cell receptors. While the complete range of their effects on glia has not been described yet, it is believed that their overall function is to inhibit adverse CNS inflammatory responses, facilitate tissue remodeling and cellular debris removal. This article summarizes the available evidence implicating the following RAMPs in CNS physiological processes and neurodegenerative diseases: cardiolipin (CL), prothymosin α (ProTα), binding immunoglobulin protein (BiP), heat shock protein (HSP) 10, HSP 27, and αB-crystallin. Studies on the molecular mechanisms engaged by RAMPs could identify novel glial targets for development of therapeutic agents that effectively slow down neuroinflammatory disorders including AD.

The excitatory effects of microRNA- 30 in the self-renewal and differentiation process of neonatal mouse spermatogonial stem cells

Background: Spermatogonial stem cells (SSCs) are considered as special stem cells since they have the ability of self-renewal, differentiation, and transferring genetic information to the next generation. Also, they considered as vital players in initiating and preserving spermatogenesis. The fate decisions of SSCs are mediated by intrinsic and extrinsic factors, among which microRNAs (miRNAs) are one of the most essential factors in spermatogenesis among endogenous regulators. However, the mechanisms by which individual miRNAs regulate self-renewal and differentiation of SSCs are unclear. The present study aimed to evaluate the impact of miRNA-30 mimic on fate determinations of SSCs. Materials and Methods: The obtained SSCs from neonatal mice (3-6 days old) were purified by MACS and flow cytometry with a promyelocytic leukemia zinc-finger marker. Then, the cultured cells were transfected with miRNA- 30 mimic, and finally, the changes in expressing ID4 and c-kit proteins were assessed by western blot analysis. Results: According to flow cytometry findings, the percentage of SSC purity was about 98.32. The expression of ID4 protein and colonization increased significantly through the transfection of miRNA-30 mimic (P<0.05). Conclusion: The miRNA-30 controls spermatogonial stem cell self-renewal and differentiation, which may have significant implications for treating male infertility. [GMJ.2020;9:e1829]

Role of Endogenous Regulators of Hem- And Lymphangiogenesis in Corneal Transplantation

Under normal conditions, the cornea, being the transparent “windscreen” of the eye, is free of both blood and lymphatic vessels. However, various diseases of the eye, like infections, can interfere with the balance between promoting and inhibiting factors, which leads to ingrowth of blood and lymphatic vessels. The newly formed lymphatic vessels increase the risk of graft rejection after subsequent corneal transplantation. Corneal transplantation is one of the most commonly performed transplantations worldwide, with more than 40,000 surgeries per year in Europe. To date, various anti-hem- and anti-lymphangiogenic treatment strategies have been developed specifically for the corneal vascular endothelial growth factor (VEGF) pathway. Currently, however, no treatment strategies are clinically available to specifically modulate lymphangiogenesis. In this review, we will give an overview about endogenous regulators of hem- and lymphangiogenesis and discuss potential new strategies for targeting pathological lymphangiogenesis. Furthermore, we will review recently identified modulators and demonstrate that the cornea is a suitable model for the identification of novel endogenous modulators of lymphangiogenesis. The identification of novel modulators of lymphangiogenesis and a better understanding of the signaling pathways involved will contribute to the development of new therapeutic targets for the treatment of pathological lymphangiogenesis. This, in turn, will improve graft rejection, not only for the cornea.

Bench to bedside review: therapeutic modulation of nitric oxide in sepsis—an update

Nitric oxide is a signalling molecule with an extensive range of functions in both health and disease. Discovered in the 1980s through work that earned the Nobel prize, nitric oxide is an essential factor in regulating cardiovascular, immune, neurological and haematological function in normal homeostasis and in response to infection. Early work implicated exaggerated nitric oxide synthesis as a potentially important driver of septic shock; however, attempts to modulate production through global inhibition of nitric oxide synthase were associated with increased mortality. Subsequent work has shown that regulation of nitric oxide production is determined by numerous factors including substrate and co-factor availability and expression of endogenous regulators. In sepsis, nitric oxide synthesis is dysregulated with exaggerated production leading to cardiovascular dysfunction, bioenergetic failure and cellular toxicity whilst at the same time impaired microvascular function may be driven in part by reduced nitric oxide synthesis by the endothelium. This bench to bedside review summarises our current understanding of the ways in which nitric oxide production is regulated on a tissue and cellular level before discussing progress in translating these observations into novel therapeutic strategies for patients with sepsis.

THE IMPACT OF THYME AND CAMOMILE FLOWER EXTRACTS ON TRYPSIN CATALYTIC ACTIVITY

The development of pathological processes in humans and animals is accompanied by an imbalance of regulatory mechanisms, in particular, proteinase enzymes and their regulators. Endogenous regulators, inhibitors and activators often fail to maintain the balance in the system “enzyme-substrate-inhibitor (activator)”, which leads to destructive inflammatory, immune reactions, and dyscrasia. Therefore, the search for unknown proteinase regulators is an important step towards the creation of new drugs to treat the diseases associated with changes in proteolysis processes. In this regard, we propose a hypothesis about the regulatory role of medicinal herbs that are widely used in both traditional and folk medicine. According to our assumption, the chemical compounds contained in medicinal herbs can both activate and inhibit proteolysis in cells. The aim of the research was to study the effects of the inflammatory process, the influence of the regulatory activity of aqueous thyme and chamomile extracts on the trypsin activity in the model experiments. Materials and Methods. In the studies the authors used crystalline trypsin, thyme, and chamomile flowers. They determined trypsin activity using the Anson method. Results. New information was obtained on the effect of an aqueous extract of medicinal herbs, in particular, thyme and chamomile flowers, on the activity of the proteolytic enzyme trypsin. This information can improve the herbs efficacy in the treatment of inflammatory processes. Conclusion. Thyme and chamomile flower extracts have an activating trypsin effect. The level of trypsin activation is proportional to the amount of extracts introduced into the incubation medium. Thyme extract has a more pronounced effect on trypsin activity. Keywords: trypsin, thyme, chamomile flowers, proteolytic activity, enzymes. Развитие патологических процессов в организме человека и животных сопровождается разбалансировкой регуляторных механизмов, в частности состояния ферментов – протеиназ и их регуляторов. Эндогенные регуляторы, ингибиторы и активаторы зачастую не справляются со своей задачей по поддержанию баланса в системе «фермент-субстрат-ингибитор (активатор)», что приводит к возникновению деструктивных воспалительных, иммунных реакций, нарушению гомеостаза. Поэтому поиск пока еще неизвестных регуляторов протеиназ представляет собой важный шаг к созданию новых препаратов для лечения заболеваний, связанных с изменениями, происходящими в процессах протеолиза. В связи с этим нами предложена гипотеза о регуляторной роли лекарственных трав, широко используемых как в традиционной, так и в народной медицине. По нашему предположению, химические соединения, содержащиеся в лекарственных травах, вероятно, могут как активировать, так и ингибировать процессы протеолиза в клетках. Целью исследования было изучение воздействия в модельных опытах воспалительного процесса, влияния регуляторной активности водных экстрактов чабреца и ромашки на активность протеолитического фермента трипсина. Материалы и методы. В исследованиях использовали «Трипсин кристаллический», чабрец и цветки ромашки. Определение активности трипсина проводили по методу, разработанному Ансоном. Результаты. Получены новые сведения о влиянии водного экстракта лекарственных трав, в частности чабреца и цветков ромашки, на активность протеолитического фермента трипсина. Данная информация позволит повысить эффективность использования лекарственных растений в лечении воспалительных процессов. Выводы. Экстракты чабреца и цветков ромашки оказывают активирующее действие на протеолитический фермент трипсин. Уровень активации трипсина пропорционален количеству экстрактов, вносимому в инкубационную среду. Экстракт чабреца оказывает более выраженное влияние на активность трипсина. Ключевые слова: трипсин, чабрец, цветки ромашки, протеолитическая активность, энзимы.