Institute of Genetics and Animal Biotechnology of the Polish Academy of Sciences

This study presents a comprehensive protocol for siRNA-mediated knockdown in the differentiation of P19 cells into neuronal-like cells. Utilizing a retinoic acid (RA)-induced neurogenesis model, P19 cells were cultured under specific conditions that facilitated the formation of embryoid bodies (EBs), which were subsequently differentiated into neuronal-like cells. In this investigation, we specifically targeted the Nfe2l1 gene using siRNA transfection to assess the efficiency and effectiveness of our protocol throughout the neuronal differentiation process. Validation of the differentiation was performed through quantitative reverse transcription PCR (RT-qPCR) analysis, measuring the expression levels of key neuronal markers, including Map2 and Pax6 along with the pluripotency marker Oct4. Additionally, the efficiency of the siRNA-mediated knockdown was confirmed by western blot analysis, which demonstrated significant gene silencing at protein levels. These findings underscore the potential of siRNA technology in elucidating gene function during neuronal differentiation and highlight the critical role of targeted gene silencing in advancing neurogenesis research. Furthermore, this study provides a robust and reliable protocol for gene knockdown in neuronal-like cells derived from P19 cells, thereby facilitating further investigations into the intricate molecular mechanisms that govern neurogenesis, neuronal maturation, and overall brain development.•Developed a novel protocol for targeted gene knockdown in P19 cells during neuronal differentiation. •Successful silencing of the Nfe2l1 gene during neuronal differentiation, validated by western blot. •This study provides a reliable protocol for gene knockdown in neuronal differentiation, aiding functional studies of genes in neurogenesis.

Diabetic retinopathy (DR) is a major complication of diabetes, leading to vision impairment and blindness. The pathogenesis of DR involves multiple factors, including hyperglycemia-induced vascular damage, hypertension, obesity, anemia, immune dysregulation, and disruption of the blood–retinal barrier (BRB). Th17 and Treg cells, two types of CD4+ T cells, play opposing roles in inflammation. Th17 cells are pro-inflammatory, producing cytokines such as IL-17A, while Treg cells help suppress immune responses and promote anti-inflammatory effects. Recent studies highlight the importance of the Th17/Treg balance in retinal inflammation and disease progression in DR. Our literature review reveals an imbalance in DR, with increased Th17 activity and reduced Treg function. This shift creates a pro-inflammatory environment in the retina, worsening vascular leakage, neovascularization, and vision loss. The limited infiltration of Treg cells suggests that Th17 cells may uniquely infiltrate the retina by overwhelming or outnumbering Tregs or increasing the expression of recruiting chemokines, rather than only taking advantage of a damaged BRB. Therapeutic strategies, such as neutralizing IL-17A and enhancing Treg function with compounds like IL-35 or curcumin, may reduce inflammation and retinal damage. Restoring the balance between Th17 and Treg cells could provide new approaches for treating DR by controlling inflammation and preventing further retinal damage.

The rigorous regulatory landscape for medical devices demands meticulous chemical characterization to ensure safety and compliance. This review examines the critical role of chemical characterization within regulatory frameworks, emphasizing its importance in the approval and market entry of medical devices. Key challenges, including the complexity of sample matrices, trace-level impurity detection, and the necessity of method validation, are thoroughly explored. In addition, the review addresses the dynamic nature of regulatory requirements, analyzing how updates in international standards, such as those from the International Organization for Standardization (ISO), the American National Standards Institute (ANSI), and the Association for the Advancement of Medical Instrumentation (AAMI), or the American Society for Testing and Materials (ASTM), shape the chemical characterization process. The review discusses future directions, including advancements in analytical technologies, the potential for increased automation and standardization, and the growing significance of managing emerging contaminants. By offering a comprehensive analysis of current practices and future trends, this review highlights the essential role of chemical characterization in ensuring the development and regulation of safe and effective medical devices.

The iron regulation mechanisms are not exactly the same between adulthood and the early postnatal period. Also, neonatal iron status is different in full-term versus preterm infants because the prenatal/gestational period, when hepatic iron accumulates, is shortened. Newborns, especially premature infants, are at high risk of iron deficiency due to inadequate iron stores, which constitute the primary source of iron to satisfy the neonate’s increasing iron requirements. In addition, frequent blood transfusions and congenital hemochromatosis may induce iron overload in the affected neonate. To understand the cause of neonatal iron deficiency/overload and to promote the development of effective therapeutic interventions in humans, different animal models have been generated by genetic engineering, low/high-iron diets, phlebotomy/transfusion, and surgical manipulation. These models use various laboratory and domestic animals to study iron imbalance. They serve as surrogate models for experiments that are ethically or practically unfeasible to conduct on human neonates. Although an animal model for studying neonatal iron disorders may not fully replicate the complexities of human diseases, it is designed to model specific aspects of these conditions. Combined data from multiple models can help offset the limitations inherent in each individual model. In this review, we outline approaches to induce neonatal iron disorders, current animal models of full-term and preterm neonates, and recommendations for diagnosis.

Background/Objectives: Observing a patient’s dietary behaviour before bariatric surgery may help to predict their diet (and indirectly, the rate of weight loss) after the procedure. Consequently, the aim of this study was to identify dietary patterns (DPs) in bariatric surgery candidates, as well as to assess the relationship between DPs, degree of obesity, and body composition. Methods: The participants were comprised of 117 bariatric surgery candidates. Data concerning their diet, lifestyle, and socioeconomic status was collected using the KomPAN® questionnaire. The following three DPs were identified using a principal component analysis: ‘Sandwiches & Sweets’, ‘Fast Food, Convenience Food & Alcohol’ and ‘Prudent’. Baseline nutritional status and body composition using electric bioimpedance were assessed. Results: Differences were found between the DPs, degree of obesity, and body composition. The Prudent DP primarily involved a high consumption of healthy products. Following the Prudent DP, differentiated the degree of obesity and the patient’s body composition the most. In turn, the Fast Food, Convenience Food & Alcohol DP was associated with a higher likelihood of Degree III obesity and a very high visceral fat level (VFL). The Sandwiches & Sweets DP included the most participants with a very high VFL. Conclusions: The bariatric surgery candidates were shown to follow different diets, and different DPs could be identified. Patients with a higher degree of obesity followed a more beneficial DP, which was likely due to their higher awareness of the risks of morbidity in obesity and of post-surgical complications. Socioeconomic factors may attenuate the association between diet and the degree of obesity and body composition in bariatric surgery candidates.

of the key disciplines that equip next‐generation researchers engaged in ethnopharmacology research with the necessary knowledge and skills to navigate the transition from traditional ethnopharmacology to modern drug discovery. image

Objectives: Resveratrol (RES) is well documented for its multiple health benefits, with a notable impact on cancer prevention and therapy. This study aimed to evaluate the effect of RES supplementation on oxidative stress in patients with head and neck cancer (HNC) receiving home enteral nutrition (HEN). Methods: This randomized, single-center, open-label study involved 72 adult patients, with 40 completing the intervention. Participants in the intervention group received 400 mg of liposomal RES daily for 12 weeks alongside HEN, while the control group received HEN only. Body composition and oxidative stress markers—including total antioxidant capacity (TAC), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), and reduced glutathione (GSH)—were measured at baseline and after 12 weeks. Results: Significant increases in TAC and SOD activity were observed in both groups. GPx activity increased significantly only in the RES group. MDA levels rose in both groups but were more pronounced in the RES group. GSH levels showed no significant changes. Phase angle (PhA) increased significantly in the RES group, while no significant change was observed in the control group. Conclusions: RES supplementation may enhance antioxidant defenses, as evidenced by increased GPx activity and improvements in TAC and SOD levels, supporting oxidative balance in patients with HNC receiving HEN. The higher MDA levels in the RES group may reflect RES’s dual antioxidant and pro-oxidant activities. Additionally, the observed increase in PhA suggests potential cellular health benefits. These findings highlight the potential of RES as a complementary antioxidant intervention in clinical oncology, warranting further investigation to clarify its therapeutic effects on oxidative stress and cellular health in cancer care.

Introduction Social media plays a significant role in fostering communities around health and wellness. The hashtag #PCOSweightloss has become a pivotal forum on the platform X, where individuals exchange experiences, share information, and motivate each other concerning weight loss strategies connected with Polycystic Ovary Syndrome (PCOS). Objective This study aims to analyze the trends of #PCOSweightloss on X to gain insights into the social media metrics, including overall hashtag reach, key themes, and potential influence on management practices of PCOS. Methods Fedica research analytics tool (https://fedica.com) was utilized to automatically evaluate quantitatively the posts on the platform X containing #PCOSweightloss from July 1st, 2017 to July 1st, 2023. Results During the defined period, 1570 users shared 12,200 posts on X, gathering 2,967,001 views. The highest number of posts originated from the United States (38.6%), followed by India (30.6%) and the United Kingdom (10.6%). Top co-occurring hashtags associated with #PCOSweightloss included #PCOS, #healthyfood, #pcosmood and #diabetes. Physicians were key contributors to the discourse, contributing to 30% of the ten most engaging posts and 20% of the top ten influencers. Sentiment analysis revealed a predominant neutral sentiment (98%), while 1% each revealed positive and negative sentiments. Conclusion Our study affirms the substantial presence of #PCOSweightloss in the PCOS discourse on X. The involvement of physicians is particularly noteworthy, as it emphasizes the importance of expert-guided discussions in the online health discourse. On the background of geographical variances and the challenge of engaging a wider audience due to the limited follower counts of many participants, there is a clear opportunity for further community engagement and outreach. The prevailing neutral tone showcases a community engaged primarily in an informational exchange, setting the stage for more profound discussions of novel PCOS weight loss strategies.

Understanding the molecular mechanisms that confer cold resistance in mammalian cells might be relevant for advancing medical applications. This study aimed to exploit the protective function of Late Embryogenesis Abundant (LEA) proteins, known to provide resistance to low temperatures in extremophiles and plants, by their exogenous expression in mammalian cells, and compare their effects with the well characterized antioxidant, vitamin E. Remarkably, the expression of LEA proteins in mammalian cells exerted cold-protective effect similar to Vitamin E. LEA proteins preserved cell viability during cold stress and ensured a normal metabolic activity after warming. Their protective action was due to the mitigation of cold-induced mitochondrial stress and the overproduction of reactive oxygen species (ROS), leading in turn to enhanced cytoskeleton stability and decreased DNA damage. Our studies showed that plant-derived LEA proteins exhibit remarkable cold protection effects in mammalian cells through their potent antioxidant properties, which was found comparable to Vitamin E.

Background: Due to their versatility, goats are becoming popular as livestock on commercial farms. To obtain a high productivity, the maintenance of homeostasis in these organisms is needed. Some supplements that could help improve their health state have been tested recently. The study examines the effects of a turmeric–rosemary extract mixture on the antioxidant status of the blood of young castrated bucks. Results: Supplementation lowered lipoperoxidation and increased TAC, probably due to the improved neutralization of reactive oxygen species (ROS) by enhanced antioxidant pathways. Despite lowering lipid peroxidation, these products can increase protein oxidation. Thus, while enhancing lipid defense mechanisms may reduce lipid peroxidation, it may also increase the level of OMP, probably due to the accumulation of lipid peroxidation metabolites. The activities of SOD, GPx, and GR remained unchanged, whereas CAT activity decreased. CP, ALT, and AST remained stable, suggesting no adverse effects on key markers of liver function. Conclusions: The high antioxidant potential observed after supplementation indicates that the turmeric–rosemary extract mixture elicited a favorable biochemical response; the supplemented compounds effectively enhanced antioxidant defenses (by improving the neutralization of ROS) without negatively affecting liver enzyme activities. Turmeric–rosemary dry extract supplementation can help with homeostasis maintenance in goat organisms and, consequently, improve their productivity.

Chronic obstructive pulmonary disease (COPD) is a prevalent chronic condition associated with substantial global morbidity and mortality. Primarily caused by prolonged exposure to harmful agents such as dust and gases, COPD is characterized by persistent airflow limitation, clinically manifesting as chronic cough, sputum production, and dyspnea. The disease course alternates between stable phases and exacerbations, with the latter often associated with pathogenic colonization of the respiratory tract. This review examines the immunological underpinnings of COPD, emphasizing the interplay between innate and adaptive immunity in disease pathogenesis. Dysregulated immune responses to environmental factors perpetuate chronic inflammation, resulting in progressive pulmonary epithelial damage and connective tissue hyperplasia, which compromise gas exchange. Exacerbations further exacerbate respiratory failure, aggravating patient symptoms and accelerating disease progression. Despite advances in COPD management, effective therapeutic options remain limited. Current treatments primarily aim to alleviate symptoms, reduce immune activation, and manage infections, yet many patients experience suboptimal outcomes. This review highlights the potential of novel therapeutic approaches targeting immune system cells and pathways. In particular, it explores the promise of oral bacterial vaccines as immunomodulatory agents to enhance immune responses and improve clinical outcomes in COPD, addressing critical gaps in current treatment paradigms.

The development of natural and eco-friendly repellents, specifically those derived from plants, has particular relevance because these plant-based repellents offer a promising alternative to chemical-based repellents. Also, plant-derived repellents could be cost-effective and safer for non-target organisms. However, the pure form of plant extracts could be hazardous as a repellent. Thus, safe concentrations and convenient delivery forms are essential. The plants of the Lamiaceae family have shown great potential in the formulation of this type of repellent against a wide range of pests, especially thanks to the properties of the components of their extracts and essential oils. This research aims to identify recent innovations in the field of plant-based repellents of the Lamiaceae family, founded on patent documents. Therefore, patent databases were searched, using appropriate keywords and refining the search using patent classification codes. We reviewed 72 patent documents, grouped into 52 simple families, published over the last ten years. The United States is the jurisdiction in which 64% of those documents were filed. The year 2019 showed the publication of the largest number of patent documents, with ten patent applications and four granted patents. We conclude that, in parallel with research in this field, which continues to advance, many innovative solutions that take advantage of the natural repellent properties of plants in the Lamiaceae family contribute to ongoing sustainable and ecological pest management efforts.

Oxidative stress is defined as a biological balance between the amount of reactive oxygen species, especially free oxygen radicals, and the action of antioxidant systems, including the so-called repair mechanisms. Disturbance of this balance causes damages in proteins, fats and nucleic acids in healthy cells. This, in turn, gives rise to many diseases, especially those that are classified as civilization diseases e.g.: diabetes, cancer, atherosclerosis, neurodegenerative processes and many others. Syringic acid (SA) is a naturally occurring phenolic compound. Numerous in vitro and in vivo studies on SA have investigated its significant impact on oxidative stress in human. This review focuses on the SA as a potential compound reducing oxidative stress parameters in civilization diseases.

Ginger ( Zingiber officinale ) has been utilized for medicinal and culinary purposes for thousands of years. Its extensive history showcases diverse uses and health benefits. In this study, we conducted a thorough analysis of the scientific literature landscape surrounding ginger using bibliometric techniques, aiming to explore recurring topics, trends, influential contributors, and main journals publishing ginger-related research. Utilizing the Web of Science (WoS) Core Collection, we identified 9,881 relevant papers published up to October 4, 2023. The analysis revealed an exponential growth in both publication and citation counts since the mid-2000s. Key phytochemical constituents, including gingerols and shogaols, emerged as focal points of research. The most frequently mentioned medical condition term identified was ‘Cancer’ (n = 577), followed by ‘Obesity’ (n = 180) and ‘COVID’ (n = 176). Journals such as the Journal of Ethnopharmacology and Food Chemistry played leading roles in sharing ginger-related articles. Our bibliometric study offers a broad overview of ginger research, highlighting a significant increase in scholarly interest globally. This analysis not only contributes quantitative insights to the existing literature but also provides a roadmap for future investigations on ginger. The scientific literature positions ginger as a promising candidate for addressing contemporary health challenges, inviting further exploration and clinical studies.

Opioids are a challenging class of drugs due to their dual role. They alleviate pain, but also pose a risk of dependency, or trigger constipation, particularly in cancer patients, who require the more potent painkillers in more advanced stages of the disease, closely linked to pain resulting from general inflammation, bone metastases, and primary or secondary tumour outgrowth-related nerve damage. Clinicians’ vigilance considering treatment with opioids is necessary, bearing in mind extensive data accumulated over decades that have reported the contribution of opioids to immunosuppression, tumour progression, or impaired tissue regeneration, either following opioid use during surgical tumour resection and post-surgical pain treatment, or as a result of other diseases like diabetes, where chronic wounds healing constitutes a challenge. During last few years, an increasing trend for seeking relationships between opioids and epithelial–mesenchymal transition (EMT) in cancer research can be observed. Transiently lasting EMT is desirable during wound healing, but in cancer, or vital organ fibrogenesis, EMT appears to be an obstacle to overcome, forcing to adjust treatment strategies that would reduce the risk for worsening of the disease outcome and patient prognosis. The same opioid may demonstrate promoting or inhibitory effect on EMT, dependently on various conditions in particular clinical cases. We have summarized current findings on this issue to uncover some rules that govern opioid-mediated EMT induction or repression; however, many aspects still remain to be elucidated.

Background Evidence indicates a wide range of andrological alterations in patients with the Severe Acute Respiratory Syndrome CoronaVirus 2 (SARS‐CoV‐2) infection and Coronavirus Disease 2019 (COVID‐19). Aim To provide an update on the andrological effects of SARS‐CoV‐2 infection and COVID‐19. Methods PubMed/MEDLINE and Institutional websites were searched for randomized clinical trials, non‐systematic reviews, systematic reviews, and meta‐analyses. Results Fifty‐four records were included from 2020 to 2024. The most representative paper categories were non‐systematic reviews ( n = 26) and systematic reviews/meta‐analyses ( n = 27). One randomized, prospective, phase 2 trial was also included. Eight topics were identified and discussed as follows: short‐ and long‐term seminal changes attributable to SARS‐CoV‐2 infection or COVID‐19; andrological effects of anti‐SARS‐CoV‐2 vaccines; the potential impact of SARS‐CoV‐2 infection and COVID‐19 on male fertility; the relationship between serum testosterone levels and COVID‐19 prognosis in men; fertility care during the pandemic; urinary/genital male system tract impairment in SARS‐CoV‐2 infection and COVID‐19; the effect of SARS‐CoV‐2 infection and COVID‐19 on circulating levels of sex steroid hormones; the impact of SARS‐CoV‐2 and COVID‐19 on sexual function and activity. Discussion SARS‐CoV‐2 can affect the whole testicular function through direct and indirect mechanisms, with a positive relation between the severity of SARS‐CoV‐2 infection and the level of deterioration of testicular function. Testicular function recovers along with the recovery from the disease. In vitro fertilization techniques ensure similar results in patients with or without previous SARS‐CoV‐2 infection or COVID‐19. Immunization with anti‐SARS‐CoV‐2 vaccines prevents andrological complaints due to naturally occurring infection. Erectile dysfunction and sexual dysfunction are frequently diagnosed in COVID‐19 patients due to several contributing factors, including hormonal imbalance and psychosocial complaints related to the pandemic.

Introduction Different Chlamydia species affect cattle and contribute to economic losses. One of them, C. pecorum , is a globally endemic livestock pathogen. Despite its endemicity, prevalence data from Poland have so far been limited. The present study aimed to obtain insight into the chlamydiae prevalence in Polish dairy cattle. Material and Methods A screening of chlamydial seroprevalence in dairy cattle was initially performed, followed by Chlamydiaceae - and species-specific real-time qPCR. Vaginal swabs (n = 239) and placenta samples (n = 2) from seropositive animals in 142 herds were collected to detect shedders. The study population consisted of cows (n = 2,780) from dairy herds (n = 1,153) located in all Polish voivodeships. Results The true animal prevalence was determined to be 33.3%, while the true herd prevalence was 42.7%. Five groups of Polish voivodeships were identified using appropriate statistical tools, highlighting differences that may arise from various factors impacting the spread of chlamydial infections. The only detected chlamydia species was C. pecorum , the presence of which was confirmed in two herds. Conclusion This study revealed that chlamydial infections are commonly present in Polish dairy cattle across the country.

Autism spectrum disorders encompass diverse neurodevelopmental conditions marked by alterations in social communication and repetitive behaviors. Advanced maternal age is associated with an increased risk of bearing children affected by autism but the etiological factors underlying this association are not well known. Here, we investigated the effects of advanced maternal age on offspring health and behavior in two genetically divergent mouse strains: the BTBR T⁺ Itpr3tf/J (BTBR) mouse model of idiopathic autism, and the C57BL/6 J (B6) control strain, as a model of genetic variability. In both strains, advanced maternal age negatively affected female reproductive and pregnancy outcomes, and perturbed placental and fetal growth, and the expression of genes in the fetal brain tissues. Postnatally, advanced maternal age had strain-dependent effects on offspring sociability, learning skills, and the occurrence of perseverative behaviors, varying between male and female offspring. These findings disentangle the relationship between genetic determinants and maternal age-related factors in shaping the emergence of autism-like behaviors in mice, highlighting the interplay between maternal age, genetic variability, and prenatal programming, in the occurrence of neurodevelopmental disorders.