Investigating the joint application of negative pressure wound treatment and tissue therapy for chronic wounds in patients with diabetes.

This study aimed to investigate the effectiveness of combined negative pressure wound therapy (NPWT) and human amniotic membrane in patients with chronic wounds associated with diabetes. A total of five patients with type 2 diabetes, including ischemic and mixed forms of diabetic foot syndrome, presenting with ischemic wounds of the lower extremities were included in this study. Patients with uncorrected limb ischemia were excluded. The treatment protocol included diabetes compensation (treatment with fractional insulin therapy), anticoagulant, metabolic therapy and angiotropic therapy, physical treatment methods, osteoporosis therapy with calcium preparations, and wound-specific interventions. The primary treatment approach involved the application of a vacuum bandage to the transplanted human amniotic membrane, which improved the adaptation of the flap to the wound surface, allowed the removal of excess wound exudate, and stimulated angiogenesis and reparative properties. The combined approach of NPWT and biotherapy was a safe and effective cure for diabetic wounds, promoting faster wound healing, reducing the need for autodermoplasty, and possibly reducing the necessity for high-level amputations.

Applied Molecular-Based Quality Control of Biobanked Samples for Multi-Omics Approach.

Biobanks are vital for high-throughput translational research, but the rapid development of novel molecular techniques, especially in omics assays, poses challenges to traditional practices and recommendations. In our study, we used biospecimens from oncological patients in Polish clinics and collaborated with the Indivumed Group. For serum/plasma samples, we monitored hemolysis, controlled RNA extraction, assessed cDNA library quality and quantity, and verified NGS raw data. Tissue samples underwent pathologic evaluation to confirm histology and determine tumor content. Molecular quality control measures included evaluating the RNA integrity number, assessing cDNA library quality and quantity, and analyzing NGS raw data. Our study yielded the creation of distinct workflows for conducting preanalytical quality control of serum/plasma and fresh-frozen tissue samples. These workflows offer customization options to suit the capabilities of different biobanking entities. In order to ensure the appropriateness of biospecimens for advanced research applications, we introduced molecular-based quality control methods that align with the demands of high-throughput assays. The novelty of proposed workflows, rooted in innovative molecular techniques, lies in the integration of these QC methods into a comprehensive schema specifically designed for high-throughput research applications.

Differentiation of neurosphere after transplantation into the damaged spinal cord.

This study aimed to compare the differentiation and survival of human neural stem/progenitor cells of various origins in vitro and after transplantation into the injured spinal cord of laboratory animals. Rats with simulated spinal cord injury were transplanted with neurosphere cells obtained by directed differentiation of HUES6 cell lines. Fluorescence microscopy was used to visualize the obtained results. HUES6#1 and iPSC#1 neurospheres showed a wide range of markers associated with glial differentiation. The expression of the proliferation marker Ki67 did not exceed 25%, both in the lines of early and late neurospheres. Although neurospheres did not fully differentiate into astrocytes in vitro, they massively approached the GFAP+ astrocyte phenotype when exposed to the transplanted environment. PSC-derived neurospheres transplanted into the site of SM injury without additional growth factors showed only moderate survival, a significant degree of differentiation into astrocytes, and moderate differentiation into neurons. The difference in the survival and differentiation of HUES6#1 and iPSC#1 neurospheres, both in vitro and in vivo, can be explained by the difference in the regulatory behavior of signaling molecules corresponding to the source of origin of PSCs. Derivatives of human PSCs of various origins obtained according to the described differentiation protocol did not mature into astrocytic populations, nor did the glycogenic transition of PSC-derived NSCs occur in vitro. The study demonstrated the impact of the injured spinal cord microenvironment on the differentiation of transplanted HUES6#1 and iPSC#1 into astrocytes. The results showed that HUES6-derived neurospheres generated 90% of GFAP+ astrocytes and 5-10% of early neurons, while iPSC-derived neurospheres generated an average of 74% GFAP+ astrocytes and 5% of early neurons in vivo.

Physiological stress level and screening for malnutrition as preoperative predictors of postoperative complications in pancreatic surgery: a retrospective study.

BACKGROUND: Assessment of 'physiological stress levels' and 'nutritional status' before surgery is important for predicting complications and indirect interventions on the pancreas. The aim of this study was to determine neutrophil-lymphocyte ratio (NLR) and nutritional risk index (NRI) indicators before surgery to predict 90-day complications and mortality in a cohort of patients with complicated chronic pancreatitis and cancer of the head of the pancreas. METHODS: We evaluated preoperative levels of NLR and NRI among 225 subjects treated at different centres located in three countries. Short-term outcomes included length of hospital stay, postoperative complications, and mortality at 90 days and were appreciated based on NLR and NRI. The level of physiological stress was divided according by the formulas: neutrophil-lymphocyte ratio (NLR) = (neutrophil count, %)/(lymphocyte count, %). The nutritional state of the patients was divided according to the INR: NRI = (1.519 × serum albumin, g/L) + (41.7 × present weight, kg / usual weight, kg)]. RESULTS: All patients were operated. An analysis of the operations performed in three institutions demonstrated mortality in chronic pancreatitis and pancreatic pseudocysts in 1.4%, in chronic pancreatitis and the presence of an inflammatory mass mainly in the pancreatic head in 1.2%, and in cancer of the pancreatic head in 5.9%. The mean preoperative NLR was normal in 33.8% of the patients, the mild physiologic stress level was 54.7%, and the moderate was 11.5% before surgery. 10.2% of patients had a normal nutritional status, 20% had mild, 19.6% had moderate, and 50.2% had severe malnutrition. In a univariate analysis, at the cutoff of NLR ≥ 9.5 (AUC = 0.803) and the cutoff of NRI ≤ 98.5 (AUC = 0.801), increasing the risk of complications was observed (hazard ratio, 2.01; 95% CI, 1.247-3.250, p = 0.006), but at the cutoff of NRI ≤ 83.55 (AUC = 0.81), we observed a survival difference in operated patients (hazard ratio, 2.15; 95% CI, 1.334-3.477, p = 0.0025). CONCLUSIONS: Our study demonstrated that NLR and NRI were predictors of postoperative complications, but only NRI was a predictor of 90-day mortality in patients after surgery.

Investigate the Possibility of Using Phosphorescence in Clinical Oncology as an Early Prognostic Test in Detecting Brain Carcinogenesis.

Phosphorescence is considered one of the non-invasive glioblastoma testing methods based on studying molecular energy and the metabolism of L-tryptophan (Trp) through KP, which provides essential information on regulating immunity and neuronal function. This study aimed to conduct a feasibility study using phosphorescence in clinical oncology as an early prognostic test in detecting Glioblastoma. This study was conducted on 1039 patients who were operated on with follow-up between January 1, 2014, and December 1, 2022, and retrospectively evaluated in participating institutions in Ukraine (the Department of Oncology, Radiation Therapy, Oncosurgery, and Palliative Care at the Kharkiv National Medical University). Method of protein phosphorescence detection included two steps. During the first step, of luminol-dependent phosphorescence intensity in serum was carried out after its activation by the light source, according to the spectrofluorimeter method, as follows. At a temperature of 30 °C, serum drops were dried for 20 min to form a solid film. After that, we put the quartz plate with dried serum in a phosphoroscope of luminescent complex and measured the intensity. With the help of Max-Flux Diffraction Optic Parallel Beam Graded Multilayer Monochromator (Rigaku Americas Corporation) following spectral lines as 297, 313, 334, 365, 404, and 434 nm were distinguished and absorbed by serum film in the form of light quantum. The monochromator exit split width was 0.5 mm. Considering the limitations of each of the non-invasive tools currently available, phosphorescence-based diagnostic methods are ideally integrated into the NIGT platform: a non-invasive approach for visualizing a tumor and its main tumor characteristics in the spatial and temporal order. Because trp is present in virtually every cell in the body, these fluorescent and phosphorescent fingerprints can be used to detect cancer in many different organs. Using phosphorescence, it is possible to create predictive models for GBM in both primary and secondary diagnostics. This will assist clinicians in selecting the appropriate treatment option, monitoring treatment, and adapting to the era of patient-centered precision medicine.

Assessment of Ecological and Toxicological State of Soils and Waters in the Neighborhood of Mining Industry Enterprises in the Armenian Highlands.

The article presents the results of the seven-year study on the state of arable lands and wastewaters in the districts of mining enterprises in Armenia. An assessment of the ecological and toxicological state of wastewaters and contaminated areas was carried out. Methods for their purification are proposed aimed at their further use and obtainment of environmentally safe agricultural products. An area of about 0.5 ha, next to the rural community of Syunik in southern Armenia, has been polluted for many years by mining sludges from the nearest mine watertight cofferdam of the Zangezur copper-molybdenum combine. Activities have been carried out to clean up the soil in this area. After plowing, soil improvers (zeolite, bentonite, and manure) were introduced into the soil. On-site treatments, soil tillage, and the introduction of soil improvers into the soil was implemented in late autumn. Soil and plant samples were taken to determine the content of heavy metals (Cu, Zn, Pb, Co, Mo, Ni). In the following spring, the area was planted with potatoes, eggplant, and peas. A rather high yield was obtained. Analysis of plant samples showed that the content of heavy metals does not exceed the permissible level of the international food safety standard. At the same time, soil studies were carried out in the adjacent areas of the Sotk mine, located to the southeast of Lake Sevan. It was revealed that due to the increase in the volume of the mining industry and the amount of rock dumps, the organoleptic and chemical indicators of the waters of the Sotk and Masrik rivers deteriorated. Compared to the previous decade, the amounts of suspended particles per 1 L of water have increased by 210…317.0%, in the waters of Sotk-by 32.1 mg/L, and in the waters of Masrik-by 13.2 mg/L. The same tendency is observed regarding the amounts of nitrates, sulfates, and hydrogen index, which is mainly due to the chemical composition of rocks. They contain a large amount of Ca, K, Na, Mg, etc. This trend is especially noticeable along the rivers, where intensive agriculture, primarily livestock farming is carried out. The material of the work solves a complex of environmental and economic problems. It is aimed at ensuring environmental safety, improving the ecological and resource properties of soils, increasing the productivity of cultural phytocenoses and the sanitary and hygienic quality of food products.

The Evaluation of Biomarkers of Physical Activity on Stress Resistance and Wellness.

Physical activity can improve health as well as reduce stress and the risk of developing several widespread diseases. However, there exists no accepted standard biomedical examination-method for stress evaluation. The purpose of this study was to investigate the effect of regular physical activity on stress and wellness as well as the evaluation of potential biomarkers in this field. This study included 105 people (mean age = 36.57 ± 1.4 years) who were randomly assigned into the exercise group 1 (EG-1) (n = 41), the exercise group 2 (EG-2) (n = 30), and the control group (CG) (n = 34). Measurements of stress and wellness were obtained by Multiscan BC-OXI before and after experimental period. This device presents a multifrequency segmental body composition 3D analyser with digital pulse oximeter. The key indicators of stress as well as for wellness were significantly improved in the EG-1. Parasympathetic activity showed significant changes as potential stress biomarker. Statistically significant gender differences were not observed in the comparable groups. The results suggest that the stress resistance and well-being significantly improved in the EG-1 due to regular physical activity. However, further research is necessary to determine effects of physical activity on integral health indicators.

Hepatic Response of Magnesium-Restricted Wild Type Mice.

Magnesium-deficiency is implicated in many metabolic disorders, e.g., type 2 diabetes and metabolic syndrome, representing risk factors for non-alcoholic fatty liver disease (NAFLD). This study aims to investigate the contribution of magnesium-restriction to the development of NAFLD. Magnesium-deficiency was induced in C57BL/6 mice by feeding a magnesium-deficient-diet. Metabolic markers as well as markers of inflammation and liver function were assessed. Furthermore, liver tissue was examined histopathologically and compared with specimens from high-fat-diet fed and control mice. Finally, the hepatic inflammatory response was quantified by determining hepatic IL-6, TNFα, and MCP-1. Magnesium-restriction resulted in at least a 2-fold significant reduction of serum magnesium levels compared to the high-fat-diet fed and control mice, whereas the hepatic magnesium content was decreased due to high-fat-diet feeding. No changes in metabolic markers in magnesium-restricted mice were observed, while the cholesterol content was elevated in high-fat-diet fed mice. Magnesium-restricted mice additionally featured inflammation and enlarged hepatocytes in liver histology. Furthermore, magnesium-restricted and high-fat-diet fed mice exhibited elevated hepatic TNFα levels compared to control mice. Accordingly, our data suggest that magnesium is involved in hepatic inflammatory processes and hepatocyte enlargement, key histological features of human NAFLD, and may therefore contribute to development and progression of the disease.

Pediatric biobanks and parents of disabled children associations opinions on establishing children repositories in developing countries.

Pediatric biobanks are an indispensable resource for the research needed to bring advances in personalized medicine into pediatric medical care. It is unclear how or when these advances in medical care may reach children, but it is unlikely that research in adults will be adequate. We conducted the screening for a hypothetic problem in various European and American pediatric biobanks based on online surveys through e-mail distribution based on the Biobank Economic Modeling Tool (BEMT) questionnaire model. Participants in the survey had work experience in biobanking for at least 3 years or more. Contact information about the survey participants was confirmed on the social networks profiles (LinkedIn), as well as on generally available websites. First, we tried creating a model which can show the pediatric preclinical and basic clinical phase relationship and demonstrate how pediatric biobanking is linked to this process. Furthermore, we tried to look for new trends, and the final goal is to put the acquired knowledge into practice, so medical experts and patients could gain usable benefit from it. We concluded that leading positions must take into account ethical and legal aspects when considering the decision to include children in the biobank collection. However, communication with parents and children is essential. The biobank characteristics influence the biobank's motives to include children in the consent procedure. Moreover, the motives to include children influence how the children are involved in the consent procedure and the extent to which children are able to make voluntary decisions as part of the consent procedure.

Overview of Cancer Control in Armenia and Policy Implications.

Cancer is the second leading cause of death in Armenia. Over the past two decades, the country has seen a significant rise in cancer morbidity and mortality. This review aims to provide up-to-date info about the state of cancer control in Armenia and identify priority areas of research. The paper analyzes published literature and local and international statistical reports on Armenia and similar countries to put numbers into context. While cancer detection, diagnosis, and treatment are improving, the prevalence of risk factors is still quite high and smoking is widespread. Early detection rates are low and several important screening programs are absent. Diagnosis and treatment methods are not standardized; there is a lack of treatment accessibility due to insufficient government coverage and limited availability of essential medicines. Overall, there is room for improvement in this sector, as research is limited and multidisciplinary approaches to the topic are rare.

Introducing multi-modal enteral medication reduced morbidity and mortality associated with necrotising enterocolitis.

AIM: Necrotising enterocolitis (NEC) is still a disease with high morbidity and mortality. The aim of the study was to analyse retrospectively whether the introduction of a multi-modal three-component enteral medication regimen resulted in a change in morbidity and mortality in neonates with NEC. METHODS: When diagnosis of NEC was established, the following multi-modal three-component enteral medication regimen was administered enterally (via nasogastric tube): an antibiotic, an antifungal agent and a probiotic. The primary outcome parameters were intestinal perforation, surgical interventions and mortality during the observational periods. RESULTS: In the study period, 2212 patients were admitted to the NICU, out of which 200 (9%) developed NEC. Significantly fewer infants died in the Intervention Group (13 of 104 infants, 13%) compared to the Control Group (38 of 96 infants, 40%) (P = .0001). No infant in the Intervention Group (0%) presented with an intestinal perforation, as compared to 15 infants (16%) within the Control Group (P = .0001). In the Control Group, 21 infants (22%) needed surgical intervention, whereas 0 (0%) infants needed this in the Intervention Group. CONCLUSION: The introduction of an enteral multi-modal three-component medication regimen resulted in a significant reduction of mortality and of need for surgical intervention in infants suffering from NEC.

Challenges and Driving Forces for Business Plans in Biobanking.

BACKGROUND: Due to increased utilization of biospecimens for research and emergence of new technologies, the availability and quality of biospecimens and their collection are coming more and more into focus. However, the long-term economic situation of biobanks is still mostly unclear. Also, the common sustainable utilization of various international biobanks is challenging due to local differences in sample processing, law and ethics. AIM: This article discusses possible strategies to achieve a sustainable utilization of biospecimens as part of the business plan of biobanks. METHODS: The following questions were addressed as part of a business plan: (1) How can a biobank build up and maintain an up-to-date infrastructure? (2) What kind of funding can support the sustainability of a biobank? (3) Is there an international solution for informed consents to enable sample and data sharing? (4) How can a biobank react during economically unstable periods? (5) Which kind of biobanking research is innovative? (6) What kind of education could be most needful for knowledge transfer in biobanking? (7) Does an expiration date for a biobank make sense according to the period of funding? CONCLUSION: A strategy for optimal utilization begins with sharing of resources, infrastructure, and investments at the planning stage of a biobank, and continues to the transfer of knowledge and know-how by education. For clinical biobanks in particular, a long-term funding and cost recovery strategy is necessary for sustainable utilization.

Magnetomitotransfer: An efficient way for direct mitochondria transfer into cultured human cells.

In the course of mitochondrial diseases standard care mostly focuses on treatment of symptoms, while therapeutic approaches aimed at restoring mitochondrial function are currently still in development. The transfer of healthy or modified mitochondria into host cells would open up the possibilities of new cell therapies. Therefore, in this study, a novel method of mitochondrial transfer is proposed by anti-TOM22 magnetic bead-labeled mitochondria with the assistance of a magnetic plate. In comparison to the passive transfer method, the magnetomitotransfer method was more efficient at transferring mitochondria into cells (78-92% vs 0-17% over 3 days). This transfer was also more rapid, with a high ratio of magnetomitotransferred cells and high density of transferred mitochondria within the first day of culture. Importantly, transferred mitochondria appeared to be functional as they strongly enhanced respiration in magnetomitotransferred cells. The novel method of magnetomitotransfer may offer potential for therapeutic approaches for treatment of a variety of mitochondria-associated pathologies, e.g. various neurodegenerative diseases.

Susceptibility of Different Mouse Wild Type Strains to Develop Diet-Induced NAFLD/AFLD-Associated Liver Disease.

Although non-alcoholic and alcoholic fatty liver disease have been intensively studied, concerning pathophysiological mechanisms are still incompletely understood. This may be due to the use of different animal models and resulting model-associated variation. Therefore, this study aimed to compare three frequently used wild type mouse strains in their susceptibility to develop diet-induced features of non-alcoholic/alcoholic fatty liver disease. Fatty liver disease associated clinical, biochemical, and histological features in C57BL/6, CD-1, and 129Sv WT mice were induced by (i) high-fat diet feeding, (ii) ethanol feeding only, and (iii) the combination of high-fat diet and ethanol feeding. Hepatic and subcutaneous adipose lipid profiles were compared in CD-1 and 129Sv mice. Additionally hepatic fatty acid composition was determined in 129Sv mice. In C57BL/6 mice dietary regimens resulted in heterogeneous hepatic responses, ranging from pronounced steatosis and inflammation to a lack of any features of fatty liver disease. Liver-related serum biochemistry showed high deviations within the regimen groups. CD-1 mice did not exhibit significant changes in metabolic and liver markers and developed no significant steatosis or inflammation as a response to dietary regimens. Although 129Sv mice showed no weight gain, this strain achieved most consistent features of fatty liver disease, apparent from concentration alterations of liver-related serum biochemistry as well as moderate steatosis and inflammation as a result of all dietary regimens. Furthermore, the hepatic lipid profile as well as the fatty acid composition of 129Sv mice were considerably altered, upon feeding the different dietary regimens. Accordingly, diet-induced non-alcoholic/alcoholic fatty liver disease is most consistently promoted in 129Sv mice compared to C57BL/6 and CD-1 mice. As a conclusion, this study demonstrates the importance of genetic background of used mouse strains for modeling diet-induced non-alcoholic/alcoholic fatty liver disease.

Sustainability in Biobanking: Model of Biobank Graz.

Research infrastructures remain the key for state-of-the-art and successful research. In the last few decades, biobanks have become increasingly important in this field through standardization of biospecimen processing, sample storage, and standardized data management. Research infrastructure in cohort studies and other sample collection activities are currently experiencing a lack of long-term funding. In this article, the Biobank Graz discusses these aspects of sustainability including the definition of sustainability and necessity of a business plan, as well as cost calculation model in the field of biobanking. The economic state, critical success factors, and important operational issues are reviewed and described by the authors, using the example of the Biobank Graz. Sustainability in the field of biobanking is a globally important matter of necessity, starting from policy making and ending with security and documentation on each operational level.

The Wide and Complex Field of NAFLD Biomarker Research: Trends.

Background. Nonalcoholic fatty liver disease is now acknowledged as a complex public health issue linked to sedentary lifestyle, obesity, and related disorders like type 2 diabetes and metabolic syndrome. Aims. We aimed to retrieve its trends out of the huge amount of published data. Therefore, we conducted an extensive literature search to identify possible biomarker and/or biomarker combinations by retrospectively assessing and evaluating common and novel biomarkers to predict progression and prognosis of obesity related liver diseases. Methodology. We analyzed finally 62 articles accounting for 157 cohorts and 45,288 subjects. Results. Despite the various approaches, most cohorts were considerably small and rarely comparable. Also, we found that the same standard parameters were measured rather than novel biomarkers. Diagnostics approaches appeared incomparable. Conclusions. Further collaborative investigations on harmonizing ways of data acquisition and identifying such biomarkers for clinical use are necessary to yield sufficient significant results of potential biomarkers.

Respiratory chain complex I is a mitochondrial tumor suppressor of oncocytic tumors.

Oncocytic tumors, also called oxyphilic tumors, are characterized by hyperproliferation of mitochondria, which histologically presents as a fine granular eosinophilic cytoplasm. In accordance with the high mitochondrial density in oncocytomas, transcript levels of subunits of complexes of the oxidative phosphorylation (OXPHOS) system are increased. Hence, for a long time oncocytomas were presumed to have a highly active aerobic mitochondrial energy metabolism. Recently, detailed analysis of all OXPHOS complexes in a variety of oncocytomas revealed loss of complex I and compensatory up-regulation of the other complexes. In half of the oncocytoma cases examined the absence of complex I is caused by disruptive mutations in mitochondrial DNA encoding complex I subunits. The new data presented here on rare oncocytomas and the accompanying review of the literature clearly indicate that complex I deficiency in combination with up-regulation of mitochondria can be regarded as a hallmark of oncocytic tumor cells. Therefore, complex I of the respiratory chain has to be added to the growing list of mitochondrial tumor suppressors.

Evaluation of risk profiles by subcutaneous adipose tissue topography in obese juveniles.

OBJECTIVE: To compare subcutaneous adipose tissue topography (SAT-top) in obese juveniles with age-matched normal-weight controls. RESEARCH METHODS AND PROCEDURES: The optical device LIPOMETER (European Patent EP 0516251) enables the non-invasive, rapid, safe, and precise measurement of the thickness of subcutaneous adipose tissue. Fifteen defined body sites (1 = neck to 15 = calf) characterize the individual SAT-top like an individual fingerprint. SAT-top of 1351 juveniles (obese: 42 boys, 59 girls, normal weight: 680 boys, 570 girls) from 7 to 19 years of age were measured. For visual comparison, the 15-dimensional SAT-top information was condensed by factor analysis into a two-dimensional factor plot. RESULTS: Both female and male obese juveniles had markedly increased adipose tissue layers at 7 = upper abdomen, 8 = lower abdomen, 5 = front chest, and 6 = lateral chest. The pubertal changes of body shape and fat distribution of the normal-weight boys and girls (boys show thinner adipose tissue layers on their legs, whereas girls had thicker adipose tissue layers at the extremities) were not seen in the obese group. Independently of age and sex, all of the obese juveniles showed a similar, more android body fat distribution with increased trunk fat. DISCUSSION: SAT-top of the obese juveniles is similar to that of patients with type 2 diabetes, polycystic ovary syndrome, and coronary heart disease. Patients with these metabolic disorders and obese juveniles are located in the factor plot in the same area. This body shape may indicate a risk profile for developing polycystic ovary syndrome (women), type 2 diabetes, and early atherosclerosis (both sexes).