Environmental manipulations generate bidirectional shifts in both behavior and gene regulation in a crossbred mouse model of extremes in trait anxiety

Although gene-environment interactions are known to significantly influence psychopathology related disease states, only few animal models cover both the genetic background and environmental manipulations. Therefore, we have taken advantage of the bidirectionally inbred high (HAB) and low (LAB) anxiety-related behavior mouse lines to generate HAB x LAB F1 hybrids that intrinsically carry both lines' genetic characteristics, and subsequently raised them in three different environments standard, enriched (EE) and chronic mild stress (CMS). Assessing genetic correlates of trait anxiety, we focused on two genes already known to play a role in HAB vs. LAB mice, corticotropin releasing hormone receptor type 1 (Crhr1) and high mobility group nucleosomal binding domain 3 (Hmgn3). While EE F1 mice showed decreased anxiety related and increased explorative behaviors compared to controls, CMS sparked effects in the opposite direction. However, environmental treatments affected the expression of the two genes in distinct ways. Thus, while expression ratios of Hmgn3 between the HAB- and LAB-specific alleles remained equal, total expression resembled the one observed in HAB vs. LAB mice, i.e., decreased after EE and increased after CMS treatment. On the other hand, while total expression of Crhr1 remained unchanged between the groups, the relative expression of HAB- and LAB-specific alleles showed a clear effect following the environmental modifications. Thus, the environmentally driven bidirectional shift of trait anxiety in this F1 model strongly correlated with Hmgn3 expression, irrespective of allele-specific expression patterns that retained the proportions of basic differential HAB vs. LAB expression, making this gene a match for environment-induced modifications. An involvement of Crhr1 in the bidirectional behavioral shift could, however, rather be due to different effects of the HAB- and LAB specific alleles described here. Both candidate genes therefore deserve attention in the complex regulation of anxiety-related phenotypes including environment-mediated effects

Bidirectional rescue of extreme genetic predispositions to anxiety: impact of CRH receptor 1 as epigenetic plasticity gene in the amygdala

The continuum of physiological anxiety up to psychopathology is not merely dependent on genes, but is orchestrated by the interplay of genetic predisposition, gene x environment and epigenetic interactions. Accordingly, inborn anxiety is considered a polygenic, multifactorial trait, likely to be shaped by environmentally driven plasticity at the genomic level. We here took advantage of the extreme genetic predisposition of the selectively bred high (HAB) and low anxiety (LAB) mouse model exhibiting high vs low anxiety-related behavior and tested whether and how beneficial (enriched environment) vs detrimental (chronic mild stress) environmental manipulations are capable of rescuing phenotypes from both ends of the anxiety continuum. We provide evidence that (i) even inborn and seemingly rigid behavioral and neuroendocrine phenotypes can bidirectionally be rescued by appropriate environmental stimuli, (ii) corticotropin-releasing hormone receptor 1 (Crhr1), critically involved in trait anxiety, shows bidirectional alterations in its expression in the basolateral amygdala (BLA) upon environmental stimulation, (iii) these alterations are linked to an increased methylation status of its promoter and, finally, (iv) binding of the transcription factor Yin Yang 1 (YY1) to the Crhr1 promoter contributes to its gene expression in a methylation-sensitive manner. Thus, Crhr1 in the BLA is critically involved as plasticity gene in the bidirectional epigenetic rescue of extremes in trait anxiety

Pharmacological prevention of SARS

The article describes the basics of prevention of acute respiratory viral diseases, as well as the most commonly used as a pharmacological prophylaxis group of drugs.В статье рассмотрены основы профилактики острых респираторных вирусных заболеваний, а также наиболее часто используемые в качестве фармакологической профилактики группы препаратов

Clinical case of mechanical and non-mechanical rhabdomyolysis complicated by acute kidney injury. Case report

To date, the problem of rhabdomyolysis remains relevant in Russia. Despite the introduction of modern methods of diagnosis, prevention and treatment, mortality remains high, up to 40%, especially with the addition of acute kidney injury. This article presents a clinical case of rhabdomyolysis, which arose as a result of the combined effects of non-traumatic (toxic effects of methadone, alcohol) and traumatic (vibration and non-physiological position during sleep) factors. This syndrome eventually led to the development of acute kidney damage up to anuria. In this clinical case, intensive infusion and detoxification therapy, including hemodialysis procedures, was carried out, which led to an almost complete restoration of the functional state of the kidneys. Thus, early diagnosis of rhabdomyolysis and AKI based on anamnesis, timely laboratory and instrumental diagnosis can prevent severe kidney damage up to terminal uremia and fatal consequences

Multicomponent coating in purulent wound healing: A randomised controlled experimental study

Background. Purulent wound healing is a pressing surgical challenge relevant in 30−35% of patient cases. To the more, wound infectious agents elaborate resistance to available drugs warranting the development of new drug combinations exserting a multidirectional effect on the wound process.Objective. Using a purulent wound model to experimentally evaluate the efficiency of a new multicomponent wound coating comprised of polyethylene oxide and carboxymethylcellulose sodium-immobilised dioxidine, methyluracil, metronidazole and lidocaine hydrochloride in comparison with a legal approved wound coating drug preparation of beeswax and propolis-based dioxidine ointment.Methods. The antimicrobial activity range (disk-diffusion method) and local anaesthetic effect (Rainier’s method) of the developed wound coating have been assessed. The healing process was studied in a purulent wound model with 72 Wistar rats divided between two equal groups. The following methods were applied: visual wound inspection (wound cleansing time, absence of wound-surrounding tissue oedema, granulation and epithelisation), planimetric parameter estimation (wound area, healing rate, wound area reduction ratio), wound contamination and pH measurement, wound section cell morphometry (granulocyte, macrophage, lymphocyte and fibroblast counts, cell index estimation). Daily dressings were applied for 15 days.Results. The developed wound coating exhibited high efficiency against Gram-positive and Gram-negative bacteria in the zone of inhibition tests. Its local anaesthetic effect was significantly superior to the approved drug by the duration of action. The wound area reduction was 94.2 (93.7; 94.8)% in the experimental group and 86 (84.2; 88.8)% in the control (differences statistically significant) already on day 10. A maximal healing rate in both groups was registered in phase 1 of the wound process being 1.4 times higher in experiment compared to the control. The wound contamination was significantly lower in experiment vs. control on day 8 (p = 0.0075). Wound pH negatively correlated with the fibroblast count and positively — with the contamination level.Conclusion. The study demonstrates high efficiency of the developed wound coating against infectious agents and its positive healing impact via reducing phase 1 and stimulating proliferation in phase 2 of the wound process

The transcription factor RUNX2 regulates receptor tyrosine kinase expression in melanoma.

Receptor tyrosine kinases-based autocrine loops largely contribute to activate the MAPK and PI3K/AKT pathways in melanoma. However, the molecular mechanisms involved in generating these autocrine loops are still largely unknown. In the present study, we examine the role of the transcription factor RUNX2 in the regulation of receptor tyrosine kinase (RTK) expression in melanoma. We have demonstrated that RUNX2-deficient melanoma cells display a significant decrease in three receptor tyrosine kinases, EGFR, IGF-1R and PDGFRβ. In addition, we found co-expression of RUNX2 and another RTK, AXL, in both melanoma cells and melanoma patient samples. We observed a decrease in phosphoAKT2 (S474) and phosphoAKT (T308) levels when RUNX2 knock down resulted in significant RTK down regulation. Finally, we showed a dramatic up regulation of RUNX2 expression with concomitant up-regulation of EGFR, IGF-1R and AXL in melanoma cells resistant to the BRAF V600E inhibitor PLX4720. Taken together, our results strongly suggest that RUNX2 might be a key player in RTK-based autocrine loops and a mediator of resistance to BRAF V600E inhibitors involving RTK up regulation in melanoma

Intraspecific Differentiation of <i>Francisella tularensis</i> Strains Using Multilocus Real-Time Polymerase Chain Reaction

The aim of the study was to develop a method for intraspecific differentiation of the tularemia microbe: subspecies tularensis (subpopulations AI and AII), holarctica (biovars japonica, EryS/R), mediasiatica, and novicida using multilocus real-time PCR. Materials and methods. We used 48 strains of F. tularensis of various subspecies, biovars, and subpopulations. Intraspecific appurtenance of the strains was carried out on the basis of the analysis of the RD-1 region variability applying PCR, the sdhA gene by Sanger fragment sequencing and by the disk diffusion method using disks with erythromycin. The selection of primers and probes was performed using the software available at www.genscript.com and GeneRunner 6.5.52. Sequence homology was assessed using the BLAST algorithm and the GenBank NCBI database. Results and discussion. New data on the structure and occurrence of the differentiation regions RD-8, RD-12, RD-28 of FTT1122c gene and its homologous sequences in strains of tularemia microbe of various subspecies have been obtained. Novel RDhm 346 bp in size, characteristic of strains of the subsp. mediasiatica, holarctica, which is deleted in subsp. tularensis and absent in subsp. novicida has been detected. Based on the detection of the FTT1670, FTT1122с, FTT1067, FTW_2084 loci, a multilocus real-time PCR has been developed – “F. tularensis 4c”, providing for identification of all subspecies of the tularemia microbe, separately for the biovar japonica of the Holarctic subspecies and subpopulations AI, AII of the subspecies tularensis. The PCR specificity was confirmed in the study of strains of tularemia microbe from the fund of the “State Collection of Pathogenic Bacteria” at the premises of the Russian Reserarch Anti-Plague Institute “Microbe”. The results obtained expand the concept of intraspecific genetic heterogeneity of tularemia microbe and possibilities of identifying the causative agent of tularemia using molecular-genetic methods. They are important for understanding the processes of adaptation of the pathogen to circulation in the host organism and environmental objects, the course of evolution and formation of new species of Francisella

Применение многокомпонентной пленки в лечении ран в эксперименте

Objectives. To optimize treatment of purulent wounds with the help of a wound coating with a multidirectional action that combines broad-spectrum antimicrobial effect, stimulation of regeneration, sorption activity and local anesthetic action. Materials and methods. The material for the study was a wound covering in the form of a film developed by the authors at Kursk State Medical University (Russian patent No. 2601897). Theexperiment was performed on laboratory animals (Wistar rats), which were divided into 2 groups (comparison and experimental), each group containing 36 animals. Purulent wound was modeled in the animals according to the method of P.I. Tolstykh. To evaluate the effectiveness of the treatment, the following methods were used: microbiological method (determination of areas of growth retardation and bacterial contamination in the wounds), Renier’s method (determination of local anesthetic activity), visual assessment of wounds, planimetric method (measurement of the wound area, percentage of area reduction and healing speed) and measurement of pH in the wounds. The statistical significance of the differences was determined with the nonparametric Mann–Whitney test. The differences were considered statistically significant at p ˂ 0.05. Results. Initially, high efficiency of the film was detected in vitro for the most common strains of test organisms, which werewound infection pathogens. The Renier index was 1.2 times higher in the film with chlorhexidine than in 2% lidocaine ointment, and the duration of general anesthesia was 25% longer. Following visual assessment of the wounds we revealed that purification and regeneration of the wounds first occurred in the animals in the experimental group; however, no statistical significance was detected. The maximal differences in the healing speed (1.6 times) were observed at 3–5 days, and the contamination of the wounds was 1.3 times lower in the experimental group than in the comparison group. According to the results of pH assessment, significant differences between the groups were noted only on day 15. Approximation of pH values to intact skin values also proved the effectiveness of treatment. Conclusion. The developed contact wound covering has high antimicrobial activity against a broad spectrum of wound infection pathogens, creates a fairly good local anesthetic effect, significantly speeds up the healing process and reduces bacterial contamination of the wound area. Thus, the developed wound covering can be recommended for further studies in the clinical setting for treatment of inflammatory processes in soft tissues.Цель. Оптимизировать лечение гнойных ран с помощью разработанного раневого покрытия с разнонаправленным действием, которое сочетает в себе противомикробный эффект широкого спектра, стимуляцию регенерации, сорбционную активность, местное анестезирующее действие. Материалы и методы. Материалом для исследования явилось разработанное нами на базе Курского государственного медицинского университета раневое покрытие в виде пленки (патент на изобретение РФ № 2601897). Эксперимент выполнен на лабораторных животных (самцы крыс породы Вистар), которые были разделены на две группы (сравнения и опытная) по 36 животных в каждой. Животным моделировалась гнойная рана по методике П.И. Толстых. Для оценки эффективности лечения применялись следующие методы: микробиологический (определение зон задержки роста и обсемененности ран), Ренье (для определения местной анестезирующей активности), планиметрический (измеряли площадь ран, долю (%) уменьшения площади и скорость заживления). Проводили визуальную оценку состояния ран и их рН-метрию. Статистическую значимость различий определяли по непараметрическому критерию Манна – Уитни. Различия считались достоверными при р ˂ 0,05. Результаты. Изначально в эксперименте in vitro была выявлена высокая эффективность разработанной пленки в отношении наиболее распространенных тест-штаммов микроорганизмов – возбудителей раневой инфекции. Индекс Ренье был в 1,2 раза выше у разработанной пленки с хлоргексидином, чем у 2%-й лидокаиновой мази, а длительность общей анестезии – на 25% дольше. На основании визуальной оценки раны показано, что очищение и регенерация ран наступали раньше у животных в опытной группе, однако статистическая достоверность различий не выявлена. Максимальные различия по скорости заживления в 1,6 раза и отмечены на сроке 3–5-е сут, а обсемененность ран была в 1,3 раза ниже в опытной группе, чем в группе сравнения. По результатам рН-метрии достоверные различия между группами выявлены лишь на 15-е сут. Приближение значений рН к значениям неповрежденной кожи также доказывала эффективность лечения. Заключение. Разработанное нами раневое покрытие обладает высокой противомикробной активностью в отношении широкого спектра возбудителей раневой инфекции, создает достаточно хороший местно-анестезирующий эффект, статистически значимо ускоряет процесс сокращения площади и обсемененности ран. Таким образом, разработанное нами раневое покрытие можно рекомендовать для дальнейших исследований в клинике для лечения гнойно-воспалительных процессов мягких тканей.

Molecular biology of breast cancer metastasis: Clinical implications of experimental studies on metastatic inefficiency

Recent technological advances have led to an increasing ability to detect isolated tumour cells and groups of tumour cells in patients' blood, lymph nodes or bone marrow. However, the clinical significance of these cells is unclear. Should they be considered as evidence of metastasis, necessitating aggressive treatment, or are they in some cases unrelated to clinical outcome? Quantitative experimental studies on the basic biology of metastatic inefficiency are providing clues that may help in understanding the significance of these cells. This understanding will be of use in guiding clinical studies to assess the significance of isolated tumour cells and micrometastases in cancer patients