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

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.

Многокомпонентное раневое покрытие в лечении экспериментальной гнойной раны

Objectives. To substantiate the effectiveness of the wound coating developed by us, which combines broad-spectrum antimicrobial effect, sorption activity, analgesic effect and prolonged action on a purulent wound.Materials and methods. The material for the study was wound coating in the form of a film developed at Kursk State Medical University (Russian patent number 2605343). Experimental animals (Wistar rats) were divided into 2 groups (comparative and experimental) of 36 animals in each. In all the experimental animals a purulent wound was modeled according to the method proposed by P.I. Tolstikh. Local anesthetic activity was determined on rabbits of the Chinchilla breed (20 individuals in each group). In the course of the study, the following methods were used: microbiological method (identification of the areas of growth retardation and wound contamination), visual assessment of the wounds, planimetric method, pH evaluation of the wounds, and Renier’s technique (for determination of the anesthetic activity). The data were processed statistically, and the statistical significance of the differences was determined using the nonparametric Mann – Whitney test. The differences were considered statistically significant at p ≤ 0.05.Results. Before the experiment on animals, high efficacy of the developed film was proven for the most common test strains of microorganisms capable of causing wound infection, using the microbiological method of investigation. Using the Renier’s method, the ability of the film to provide a local anesthetic effect was demonstrated, which was significantly higher than that of the 2% lidocaine ointment. Following visual evaluation of the wound, it was found that purification and regeneration of the wounds occurred earlier in the animals from the experimental group. The speed of wound healing in the experimental group was 1.3 times higher at 1–3 days, whereas at 5–8 days the situation was reversed in favor of the comparison group. In addition, at day 8 wound contamination in the experimental group was 1.2 times lower than in the comparison group. According to the results of the pH evaluation, significant differences between the groups were noted at days 5, 8, 15. Approximation of pH values to those of intact skin also proved the effectiveness of using the developed film.Conclusion. The results of the conducted studies confirmed broad-spectrum antimicrobial activity of the developed film, its local anesthetic activity and efficacy in treatment of experimental purulent wounds, which allows to recommend it for further preclinical trials. Цель. Обосновать эффективность применения разработанного нами раневого покрытия, которое сочетает в себе противомикробный эффект широкого спектра, сорбционную активность, обезболивающее влияние, пролонгированное действие, на модели гнойной раны в эксперименте.Материалы и методы. Материалом для исследования послужило раневое покрытие в виде пленки, разработанное на базе Курского государственного медицинского университета (патент на изобретение РФ № 2605343). Экспериментальные животные (крысы породы Вистар) были распределены на две группы (сравнения и опытная) по 36 животных в каждой. Всем подопытным животным моделировалась гнойная рана по методике П.И. Толстых. Местно-анестезирующую активность определяли на кроликах породы шиншилла (по 20 особей в каждой группе). В ходе исследования применялись следующие методы: микробиологический (определение зон задержки роста и обсемененности ран), визуальная оценка состояния ран, планиметрический, рН-метрия ран, метод Ренье (для определения анестезирующей активности). Данные обработаны статистически, достоверность отличий определяли по непараметрическому критерию Манна – Уитни. Различия между группами считались статистически значимыми при р ≤ 0,05.Результаты. До начала эксперимента на животных на основании микробиологического метода исследования была показана высокая эффективность разработанной пленки в отношении наиболее распространенных тест-штаммов микроорганизмов, способных вызывать раневую инфекцию. Также на основании метода Ренье показана ее способность оказывать местно-анестезирующее действие, достоверно превосходящее по эффективности 2%-ю лидокаиновую мазь. На основании визуальной оценки раны выявлено, что очищение и регенерация ран наступали раньше у животных в опытной группе. Скорость заживления ран в опытной группе была в 1,3 раза выше на сроке 1–3-и сут, а на 5–8-е сут ситуация была обратной в пользу группы сравнения. Кроме того, обсемененность ран в опытной группе была в 1,2 раза ниже на 8-е сут, нежели в группе сравнения. По результатам рНметрии достоверные различия между группами были отмечены на 5-, 8-, 15-е сут. Приближение значений рН к значениям неповрежденной кожи также доказывала эффективность применения разработанной пленки.Заключение. Результаты проведенных исследований подтвердили широкий спектр противомикробного действия разработанной пленки, ее местно-анестезирующую активность, эффективность при лечении экспериментальных гнойных ран, что позволяет рекомендовать ее для дальнейших доклинических испытаний.

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

The clinical case of limb-girdle muscle dystrophy 2Q associated with myasthenic syndrome and lung damage

Limb-girdle muscle dystrophy 2Q is one of the rarest forms of plectinopathies and is represented by an isolated muscular dystrophic syndrome, according to two previously described literature reports. There are five forms of plectinopathies, including limb-girdle muscle dystrophy 2Q, are caused by mutations in the PLEC gene, the alternative splicing of which determines the synthesis of 9 isoforms of the plectin protein (1, 1a, 1b, 1c, 1d, 1e, 1f, 1g, 3) performing cytolinker function in the neuronal, epithelial and muscle tissue.The article describes the family observation of three sick siblings with the limb-girdle muscle dystrophy 2Q phenotype due to the presence of a new homozygous mutation (NM_201378.3:c.58G&gt;T, NP_958780.1:p.Glu20Ter) in the isoform 1f PLEC revealed by whole-exome sequencing. Clinical, electromyography, visualization and histopathological features of limb-girdle muscle dystrophy 2Q were analyzed in detail. The onset of clinical manifestations in all the described siblings was observed in early childhood with moderate weakness mainly in the pelvic girdle muscles and proximal lower limbs with minimal involvement of the muscles of the shoulder girdle. A distinctive aspect is the stagnation of the myodystrophic process until 20—21 years, followed by the progression and development of episodes of respiratory failure, as well as the formation of rigidity of the cervical, thoracic spine and moderate contracture of the Achilles tendons. Typical features are marked atrophy of paravertebral muscles with the formation of pterygoid scapula and the presence of hypertrophy m. gastrocnemius, m. quadriceps femoris, m. deltoideus and m. triceps brachii. Histopathological examination m. vastus lateralis revealed myodystrophic process without inflammatory infiltration, muscle fiber cytoskeleton disorganization resulted from the plectin loss.Electrocardiography signs of the early repolarization syndrome, focal cardiosclerosis and sinus tachycardia are described. For the first time, involvement in the pathological process of pulmonary tissue in the form of noninfectious bronchiolitis, atelectasis, and the development of the myasthenic syndrome causing episodes of respiratory failure resulted in the death of two described siblings aged 29 and 31 years. Discussed pathogenetic role of PLEC 1f isoform in the development of described syndromes, expands understanding of rare nosology limb-girdle muscle dystrophy 2Q

Metastasis Suppressors and the Tumor Microenvironment

The most dangerous attribute of cancer cells is their ability to metastasize. Throughout the process of metastasis, tumor cells interact with other tumor cells, host cells and extracellular molecules. This brief review explores how a new class of molecules – metastasis suppressors – regulate tumor cell–microenvironmental interactions. Data are presented which demonstrate that metastasis suppressors act at multiple steps of the metastatic cascade. A brief discussion for how metastasis suppressor regulation of cellular interactions might be exploited is presented

Новая мутация в гене TYMP: клинико-морфологическая характеристика пациента с синдромом MNGIE

Mitochondrial neurogastrointestinal encephalomyopathy is an extremely rare (1–9:1 000 000, Orphanet, 2021) multisystem genetic disease caused by mutations in the TYMP gene encoding the enzyme thymidine phosphorylase.The article presents the data of a thirteen‑year survey on 40‑year‑old patient D. with clinical manifestations of mitochondrial neurogastrointestinal encephalomyopathy syndrome associated with the previously undescribed missense mutation c.1301G&gt;T (p.Gly434Val) of the TYMP gene. Detailed clinical picture (gastrointestinal dysfunction, cachexia, blepharoptosis, ophthalmoparesis, peripheral polyneuropathy and leukoaraiosis), electroneuromyography data (demyelination with secondary axonopathy), high blood serum level of dihydrothymine together with normal levels of thymidine and deoxyuridine made it possible to verify the diagnosis. Histopathological examination revealed atrophy of the longitudinal (outer) muscle layer of the small and large intestines and a significant decrease in the number of CD117+ cells (telocytes), signs of damage to the striated skeletal muscles of a mixed nature with a predominance of the myogenic pattern, as well the destruction of the myelin sheaths of peripheral nerves. Histochemical examination did not reveal “ragged red fibers” characteristic of mitochondrial pathology. Transmission electron microscopy demonstrated the presence of megalomitochondria in the myocardium.Синдром митохондриальной нейрогастроинтестинальной энцефаломиопатии – редкое (1–9:1000000, Orphanet, 2021) генетическое мультисистемное заболевание, обусловленное мутациями в ядерном гене TYMP, кодирующем фермент тимидинфосфорилазу.Представлены данные 13‑летнего наблюдения пациентки Д., 40 лет, с синдромом митохондриальной нейрогастроинтестинальной энцефаломиопатии, связанным с ранее не описанной миссенс‑заменой c.1301G&gt;T (p.Gly434Val) в гене TYMP. Диагноз синдрома митохондриальной нейрогастроинтестинальной энцефаломиопатии был поставлен на основании клинических проявлений (дисфункция желудочно‑кишечного тракта, кахексия, блефароптоз, офтальмопарез, периферическая полинейропатия и лейкоэнцефалопатия), результатов электронейромиографии (демиелинизация с вторичной аксонопатией), а также повышения уровня дигидротимина в сыворотке крови при нормальных уровнях тимидина и дезоксиуридина. Патогистологическое исследование выявило атрофию продольного (наружного) мышечного слоя тонкой и толстой кишок и значимое уменьшение количества CD117+‑клеток (телоцитов), поражение скелетных мышц смешанного характера с преобладанием миогенного паттерна, а также деструкцию миелиновых оболочек периферических нервов. Исследование S100‑положительных вегетативных образований кишечной стенки не выявило патологических изменений. При гистохимическом исследовании не были обнаружены «рваные красные волокна», характерные для митохондриопатий. Трансмиссионная электронная микроскопия продемонстрировала наличие полиморфизма митохондрий кардиомиоцитов и мегаломитохондрий лейомиоцитов кишечника

Molecular biology of breast cancer metastasis: Genetic regulation of human breast carcinoma metastasis

The present is an overview of recent data that describes the genetic underpinnings of the suppression of cancer metastasis. Despite the explosion of new information about the genetics of cancer, only six human genes have thus far been shown to suppress metastasis functionally. Not all have been shown to be functional in breast carcinoma. Several additional genes inhibit various steps of the metastatic cascade, but do not necessarily block metastasis when tested using in vivo assays. The implications of this are discussed. Two recently discovered metastasis suppressor genes block proliferation of tumor cells at a secondary site, offering a new target for therapeutic intervention