Environmental pollution with heavy metals has been accentuated by anthropogenic activities and constitute a major cause of ecosystem degradation and public health concern. Environmental reclamation assisted by plants and associated microbial communities is gaining momentum as an alternative to traditional physicochemical measures. This bioremediation technique offers several environmental benefits, notably improvement of soil fertility, promotion of ecosystem's biological diversity, and maintaining of aesthetics and health of landscapes. However, bioaccumulation of toxic elements in plant tissues can alter plant metabolism, destructs growth and yield, generates oxidative stress, and causes damage to DNA and membranes, etc.Beneficial rhizobacteria, known as plant growth promoting bacteria (PGPR), establish a profitable relationship with plants and promote their growth under wide stress conditions. The mechanisms by which PGPR support plant growth can be categorized as direct  and indirect (such as diminution of plant stress based on ACC deaminase activity and induction of systemic resistance). Besides, PGPR encompass species exhibiting biocontrol activities through the secretion of siderophores, antimicrobials, the inhibition of communication signals in the rhizosphere, etc. The ability of PGPR to colonize plant roots and form a robust and well-established biofilm also determines the effectiveness of microbial inoculants.In this regard, it is known that metallo-tolerant PGPR assist the establishment and development of plants in metal-polluted environments. In addition, they can still promote plant growth and alleviate metal stress, which enhances the phytoremediation capacity of plants. Bacteria have evolved multiple resistance mechanisms to counteract the metal toxicity, including oxidation/reduction, extrusion from the cell through specific efflux pumps, complexation, precipitation, volatilization, etc., in order to survive in the harsh environments.For this approach to be useful, metal-tolerant bacterial strains well adapted to particular regions must be employed. In this context, the study of phytomicrobiomes associated with plants is one of the most relevant areas of research. Omics technologies open new and exciting possibilities in the application of microbe-assisted phytoremediation by identifying, not only new bacterial strains with metal resistance and PGP traits, but also by providing comprehensive data concerning their effect on plants. This Research Topic is a compilation of updated research that explains the role of PGPR in the reclamation of metal-polluted soils.Abid et al. examined the synergistic utilization of a hyperaccumulator plant, Sedum alfredii, and the cadmium-resistant Bacillus sp. M6, for Cd phytoremediation in soils. The bacterium improved the Cd phytoextraction capacity. Moreover, the addition of some amendments such as biochar, rhamnolipids, and glutamic acid further improved Cd mobilization and extraction. On the other side, the rhizosphere's metagenomics analysis showed changes in the rhizospheric bacterial populations upon Bacillus inoculation particularly in the relative abundance of ammonia-oxidizing prokaryotes and the cadA gene responsible for Cd extrusion from bacterial cells.Zhang et al. deals with Cd phytoremediation, using a different Cd hyperaccumulator, Solanum nigrum. The authors showed that inoculation with the Cd-tolerant strain Bacillus sp. PGP15, which displayed several plant growth promoting abilities, ameliorated plant growth and alleviated Cd stress. Moreover, full genome sequencing identified some transposable elements putatively involved in the evolution of heavy metal resistance in this bacterium.The work by Mghazli et al. investigated the revegetation of phosphate mine wastes particularly in semi-arid regions. This study focused on the isolation, characterization, and identification of indigenous bacterial strains from Moroccan phosphate mining wastes. After testing the PGP activities of the whole collection, the authors selected the best-performing bacteria Stenotrophomonas rhizophila which was able to improve plant growth and chlorophyll content. In addition, this study is the first to determine the PGP activity of Brevibacterium anseongense.Sun et al. conducted a bibliometric review of scientific literature concerning rhizosphere microorganisms uncovering data from the last decade (2012–2022). The results have shown that the interest in this area is increasing which is evident from the rapidly increasing number of publications in many countries. Furthermore, cooperation networks between them are revealed, indicating high collaboration at the global level in this Research Topic.Finally, In conclusion, we would like to emphasize that research on PGPR is very advanced at the academic level, and yet, the acceptance and actual field application of biofertilizers are still far from complete realization. Furthermore, differences can be observed in the application of PGPR amongst countries, both for crop optimization  and environmental management. Estimation of the future market for biofertilizers depicts a promising scenario but requires the attention of the global scientific community and agro-biotechnology companies to fill the gaps between research and real practitioners. In this context, it is important to design useful and region-specific inoculants that could adapt well to particular stressful environmental conditions in order to develop what are called “tailored phytoremediation projects.” Furthermore, the development of local biotechnology companies for the introduction and popularization of this technology among farming communities in the near future could be a source of qualified jobs, generate opportunities, and foster social and scientific development. In addition, recently the use of several agricultural and/or industrial wastes as amendments, together with PGPR, is being proposed to improve and make phytoremediation profitable. This approach is important in the current scenario of the circular economy.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.


Hymenoplasty is a reconstructive surgical procedure aimed at repairing the hymen, and it has gained significant attention in the Indian context due to cultural and societal factors associated with virginity. This review article aims to investigate contemporary patterns of hymenoplasty in India and assess their influence on young adults. The review employed a systematic approach to gather and analyze relevant literature from various academic databases. Selection criteria were established to ensure the inclusion of high-quality studies focusing on hymenoplasty in India. The review provides an overview of the historical context of hymenoplasty, highlighting cultural and societal factors that influence the practice. It also examines the evolution of attitudes and perceptions regarding virginity and the hymen in India. Contemporary trends in hymenoplasty are analyzed, including the prevalence and frequency of procedures, demographic factors, patient profiles, popular motivations, and variations in surgical techniques. The socio-cultural impact on young adults is explored, emphasizing the psychological and emotional implications, the influence of societal norms and family expectations, gender dynamics, and the stigma and societal judgment that individuals face. The article concludes with recommendations for future research, including evaluating long-term outcomes and psychological well-being, exploring cultural and regional variations, assessing counseling services, and further examining ethical considerations and professional guidelines. Furthermore, the importance of comprehensive sex education, open dialogue, and discussions regarding virginity and sexual health are highlighted as crucial steps toward creating a more informed and empathetic society. Hymenoplasty is a surgical procedure that reconstructs or repairs the hymen, a thin membrane located at the entrance of the vaginal canal. Traditionally associated with virginity, hymenoplasty has gained significant attention in recent years due to its cultural, social, and psychological implications. While the procedure is not exclusive to India, it holds particular significance in the Indian context, where cultural and societal norms surrounding virginity are deeply ingrained .India is a diverse country with a rich cultural heritage where concepts of purity, honor, and virginity are important. The association of the hymen with virginity and societal expectations can exert tremendous pressure on young individuals. Hymenoplasty has emerged as a response to these expectations, offering a way to reconcile personal choices with societal norms ,3.Understanding the contemporary trends and impact of hymenoplasty in India is crucial for several reasons. First, it sheds light on the evolving dynamics of sexuality, gender, and identity in a rapidly changing society. Second, it highlights the influence of cultural and social factors on individual decision-making and mental well-being. Lastly, studying hymenoplasty can contribute to broader conversations about reproductive rights, bodily autonomy, and the need for comprehensive sexual education -6.This review article provides a comprehensive analysis of contemporary trends and the impact of hymenoplasty on young adults in India. This review examines existing literature and research to explore the motivations, practices, and consequences associated with hymenoplasty. Through this review, we hope to contribute to a better understand of what is a complex social and medical phenomenon. By addressing the significance and implications of this practice, we aim to facilitate informed discussions, raise awareness, and provide insights for healthcare providers, policymakers, and society at large.MethodologyThis comprehensive review employed a systematic approach to gather and analyze relevant literature on hymenoplasty in the Indian context. The research encompassed qualitative and quantitative research studies and academic papers to ensure a comprehensive understanding of the subject matter. An extensive search was conducted across multiple academic databases, including PubMed, Scopus, and Google Scholar, to identify relevant sources. The search utilized pertinent keywords and phrases such as "hymenoplasty," "virginity surgery," "reconstructive surgery," and "India." By employing this systematic research approach, a wide range of scholarly materials was accessed and reviewed to provide a comprehensive overview of the topic. Maintaining the rigor and relevance of the review necessitated the establishment of specific selection criteria for the inclusion of studies. These criteria encompassed several key aspects. Firstly, studies conducted in India or focusing specifically on hymenoplasty in the Indian context were prioritized. Additionally, preference was given to studies published in peer-reviewed journals, conference proceedings, and reputable sources. Studies published within a specified timeframe were considered to ensure the incorporation of recent research. Furthermore, studies that provided insights into contemporary trends, motivations, impact, and ethical considerations of hymenoplasty were included. Lastly, studies in English or with comprehensive English summaries were preferred to facilitate analysis and interpretation. By adhering to these selection criteria, the review ensured the inclusion of high-quality research that directly addressed the study's objectives. Figure Historical context of hymenoplasty in IndiaOverview of Cultural and Societal Factors Influencing HymenoplastyHymenoplasty does not exist in a vacuum but is deeply intertwined with cultural and societal factors in India. Understanding these influences is crucial to comprehend the motivations behind hymenoplasty. In Indian culture, virginity is traditionally associated with purity, honor, and societal expectations, particularly for women. The intactness of the hymen has been considered a symbol of a woman's chastity and virtue, often linked to concepts of family honor and marital suitability. These cultural norms, combined with the influence of religious beliefs and social pressures, contribute to the demand for hymenoplasty to conform to societal expectations and preserve one's perceived purity ,7,8.Evolution of Attitudes and Perceptions Regarding Virginity and Hymen in IndiaOver time, attitudes and perceptions regarding virginity and the hymen have evolved significantly in India. With the modernization of society and increased exposure to global influences, traditional notions of virginity have faced challenges and scrutiny. Discussions around sexual liberation, individual autonomy, and gender equality have gradually emerged, evaluating the significance placed on the hymen as the sole determinant of a woman's virtue. The younger generation, in particular, is more likely to question and challenge these societal norms, seeking to redefine notions of virginity and navigate a more nuanced understanding of sexuality ,10.However, despite these evolving perspectives, the cultural and societal expectations around virginity remain deeply rooted in many communities across India . The preContemporary trends in hymenoplasty in IndiaCurrent Prevalence and Frequency of Hymenoplasty ProceduresExamining the current prevalence and frequency of hymenoplasty procedures in India provides valuable insights into the contemporary trends surrounding this practice. Recent studies and reports suggest that the demand for hymenoplasty has steadily increased in India. Individuals from diverse backgrounds, including urban and rural areas, across different country regions, seek the procedure. Understanding the scale and extent of hymenoplasty in contemporary India helps contextualize its societal impact and shed light on the prevalence of societal expectations surrounding virginity ,14.Analysis of Demographic Factors and Patient ProfilesAnalyzing demographic factors and patient profiles associated with hymenoplasty reveals important patterns and trends. This analysis may include age, education, socioeconomic status, and geographic location. It helps identify groups or communities more likely to seek hymenoplasty, providing a deeper understanding of the factors influencing this decision. Exploring demographic factors and patient profiles assists in identifying the social, cultural, and economic contexts in which hymenoplasty is more prevalent and offers insights into the intersectionality of individuals seeking this procedure .Popular Motivations and Reasons for Undergoing HymenoplastyUnderstanding the motivations and reasons behind individuals' decisions to undergo hymenoplasty is essential for comprehending societal and personal factors. Various motivations can drive individuals to seek hymenoplasty, including societal pressures, cultural expectations, desires for marriageability, and the need to preserve family honor. Exploring these motivations helps reveal the complex interplay between personal agency, cultural influences, and societal norms. Additionally, examining the motivations and reasons can contribute to discussions on gender roles, sexual autonomy, and the impact of societal expectations on individuals' choices .Variations in Surgical Techniques and ProceduresHymenoplasty procedures can vary in techniques and approaches. Analyzing the variations in surgical techniques provides insights into the medical aspects of hymenoplasty and the advancements in reconstructive surgery . DiffereSocio-cultural impact on young adultsPsychological and Emotional Implications for Young AdultsThe socio-cultural impact of hymenoplasty on young adults encompasses significant psychological and emotional implications. The pressure to conform to societal expectations of virginity can lead to anxiety, guilt, and shame for individuals considering or undergoing hymenoplasty. Young adults may experience various emotions, including fear of judgment, internal conflict, and a sense of dissonance between personal desires and societal norms. Understanding the psychological and emotional implications is crucial for providing appropriate support and counseling to young adults navigating these complex dynamics ,18.Influence of Societal Norms, Traditions, and Family ExpectationsSocietal norms, traditions, and family expectations are pivotal in shaping the decision-making process for young adults considering hymenoplasty. In many communities in India, the perception of virginity as a marker of honor and respectability can exert immense pressure on individuals. Young adults often find themselves torn between personal desires and the desire to meet societal expectations. The weight of cultural and family traditions can significantly influence the decision to undergo hymenoplasty, highlighting the need to critically examine the impact of these influences on individual autonomy and agency ,20.Gender Dynamics and the Role of Patriarchy in Decision-MakingGender dynamics and the role of patriarchy are crucial aspects of the socio-cultural impact of hymenoplasty on young adults. Hymenoplasty is deeply intertwined with notions of female sexuality, control, and ownership of women's bodies. The decision to undergo hymenoplasty is often shaped by patriarchal structures that place a premium on women's virginity, reinforcing power imbalances and limiting women's autonomy. Examining gender dynamics and the influence of patriarchy is essential in understanding the societal forces that shape young adults' choices and their implications for gender equality and women's empowerment .Stigma, Shame, and Societal Judgment Faced by Young AdultsYoung adults considering or undergoing hymenoplasty often face stigma, shame, and societal judgment. Society's scrutiny and the fear of being labeled as "impure" or "immoral" can be daunting for individuals. The stigma surrounding hymenoplasty can have far-reaching consequences, including social ostracism, strained relationships, and psychological distress. Understanding the stigma and societal judgment faced by young adults is critical for fostering empathy, promoting inclusivity, and challenging the societal norms perpetuating the judgment and discrimination against individuals seeking hymenoplasty .Ethical considerations and legal frameworkExamination of Ethical Concerns Surrounding HymenoplastyThe practice of hymenoplasty raises several ethical concerns that warrant careful consideration. One primary ethical concern revolves around the notion of informed consent. It is crucial to ensure that individuals seeking hymenoplasty fully understand the procedure's risks, benefits, and potential outcomes. The ethical implications of surgical intervention based on societal expectations and pressures must be critically evaluated ,24.Another ethical concern is the role of healthcare professionals in providing hymenoplasty services. Medical practitioners must navigate the tension between respecting patients' autonomy and upholding ethical principles such as beneficence and non-maleficence. The ethical obligations of healthcare professionals extend beyond the physical aspects of the procedure and encompass the psychological well-being and long-term consequences for patients ,25. AddiLegal Regulations and Policies Governing Hymenoplasty in IndiaIn India, legal regulations and policies regarding hymenoplasty are important to consider. Currently, there are no specific laws addressing hymenoplasty directly. However, the procedure may fall under the broader framework of medical ethics, patient rights, and professional conduct regulations ,23,26.Medical professionals are guided by ethical guidelines and codes of conduct established by medical associations and regulatory bodies. These guidelines emphasize the importance of informed consent, patient confidentiality, and healthcare professionals' responsibility to prioritize their patient's best interests . It is ePsychological support and counselingImportance of Pre- and Post-Operative Psychological SupportPsychological support is crucial for individuals considering and undergoing hymenoplasty before and after the procedure. Pre-operative psychological support helps individuals make informed decisions by providing a safe space to explore their motivations, expectations, and concerns. It allows for an open and non-judgmental dialogue, enabling individuals to reflect on the societal pressures, cultural influences, and personal factors influencing their decision .Post-operative psychological support is equally important as individuals navigate the emotional and psychological aftermath of the procedure. It helps individuals process their experiences, manage any feelings of guilt or shame, and address potential challenges that may arise, such as societal judgment or strained relationships. Post-operative psychological support can contribute to empowerment, self-acceptance, and improved emotional well-being .Role of Counseling Services in Managing Expectations and Psychological Well-BeingCounseling services are crucial in managing expectations and promoting psychological well-being for individuals considering or undergoing hymenoplasty. Counseling provides a supportive and non-judgmental environment where individuals can express their concerns, explore their motivations, and gain a deeper understanding of the implications of their decision .Counselors can help individuals develop realistic expectations about the outcomes of hymenoplasty, ensuring that they understand the limitations and potential risks involved. They can also address misconceptions and myths surrounding virginity, challenging societal norms and promoting a more nuanced understanding of sexuality .Furthermore, counseling services can assist individuals in building resilience, coping mechanisms, and self-esteem, helping them navigate the potential stigma and societal judgment they may face. Counseling sessions may involve techniques such as cognitive-behavioral therapy, supportive counseling, and psychoeducation to address the decision-making process's psychological and emotional aspects and the procedure's aftermath . The rolFuture directions and recommendationsAreas for Further Research and InvestigationWhile significant progress has been made in understanding hymenoplasty in the Indian context, several areas warrant further research and investigation. Some potential areas for future research include the following.Long-term outcomes and psychological well-being: It is important to conduct research that focuses on the long-term psychological impact of hymenoplasty on individuals. This research should explore factors such as self-esteem, body image, and sexual satisfaction to understand better how hymenoplasty affects individuals' overall well-being beyond the immediate post-operative period. Longitudinal studies that follow individuals over an extended period can provide valuable insights into the psychological implications and potentially long-lasting effects of hymenoplasty ,29.Cultural and regional variations: Investigating the cultural and regional variations in attitudes, motivations, and prevalence of hymenoplasty can enhance our understanding of this practice within different communities and geographic locations in India. By exploring how cultural norms, traditions, and societal expectations influence the demand for hymenoplasty, we can gain insights into the diverse factors contributing to the decision-making process. This research can also shed light on the impact of regional socio-cultural factors on the prevalence and acceptance of hymenoplasty, allowing for targeted interventions and education efforts .Evaluation of counseling and support services: Research should assess the effectiveness of counseling and support services in assisting individuals before and after hymenoplasty. This evaluation should go beyond satisfaction surveys and measure outcomes such as emotional well-being, empowerment, and the ability to make informed decisions. Understanding the impact of counseling and support services can guide the development of evidence-based interventions that address the unique needs of individuals considering or undergoing hymenoplasty. This research can also inform the training and guidelines for healthcare professionals, ensuring they are equipped to provide comprehensive care that addresses the psychological and emotional aspects of hymenoplasty .Ethical considerations and professional guidelines: Further exploration of the ethical concerns surrounding hymenoplasty and developing professional guidelines are essential to navigate this procedure's complex ethical landscape. Research should delve into the ethical implications of performing hymenoplasty, including issues of informed consent, patient autonomy, and the objectification of women's bodies. Examining the existing ethical guidelines and regulations and engaging in ethical discussions can enhance the ethical framework for healthcare professionals involved in hymenoplasty. This research can also contribute to developing comprehensive professional guidelines that promote ethical practices and patient-centered care .Advocacy for Comprehensive Sex Education and AwarenessPromoting comprehensive sex education and awareness is crucial in addressing the underlying issues surrounding hymenoplasty. By incorporating comprehensive sex education in schools and educational institutions, young adults can gain accurate information about virginity, sexuality, and reproductive health. This education should promote healthy attitudes toward sexuality, challenge harmful societal norms, and foster respect for individual choices ,36. AdvoImportance of Open Dialogue and Discussions Regarding Virginity and Sexual HealthEncouraging open dialogue and discussions surrounding virginity and sexual health is crucial for dismantling societal taboos and promoting a more inclusive and understanding society. Creating safe spaces for conversations, both online and offline, can facilitate a deeper understanding of the complexities surrounding hymenoplasty, foster empathy, and challenge societal judgments ,14,35. BThe findings of this review have important implications for healthcare providers, policymakers, and society. Healthcare providers should recognize the psychological and emotional implications of hymenoplasty and provide comprehensive care that includes pre- and post-operative psychological support. They should also adhere to ethical guidelines and foster open communication with patients to ensure informed consent and patient-centered care. Policymakers can use the findings to develop regulations and policies that protect the rights of individuals seeking hymenoplasty while addressing ethical concerns and ensuring quality healthcare services. Advocacy for comprehensive sex education and awareness can help challenge societal norms and promote a more inclusive and informed approach to sexual health. Society should engage in open dialogue and discussions surrounding virginity, sexual health, and the complexities of hymenoplasty. By promoting empathy, understanding, and respect for individual choices, we can create a more supportive and accepting environment for individuals navigating the decision-making process and the aftermath of hymenoplasty.


Neurological disorders are the major cause of disability-adjusted life years (DALYs) and the second leading cause of death worldwide GBD, . These dThe objective of this Research Topic is to compile a collection of scholarly articles that offers insight into the spinal and supraspinal circuits of patients with neuromuscular disorders. These articles should present various computational, theoretical, and experimental models to aid in the effective management of neuromuscular disorders, such as strokes, using EMG-based rehabilitative devices. Therefore, we invited researchers from the field of biomedical engineering, biomechanics, neuroscience, physical therapy, etc. to contribute to this issue. In response to the call for contributions, three articles and a brief research report were submitted and accepted. We have outlined a summary of each article published in this Research Topic below.Seo et al. decomposed multichannel EMG signals from the upper extremity to examine motor modules' contribution to exploratory isometric forces among stroke patients during a three-dimensional target force task. The motor modules also known as muscle synergies are the building blocks of movement, and the CNS selects a small dimension of motor modules from a high-dimensional space to form a movement  and hand  muscles compared to the control group. In particular, the stroke group displayed a significantly smaller hand aperture compared to the control group during shoulder abduction or elbow extension movement. This constrained functional mechanical behavior observed in stroke patients was due to the co-contraction of hand muscles, which is caused by increased neural coupling between the shoulder and hand muscles. This phenomenon is revealed as high inter-muscular coherence in hand muscles, as computed from the sEMG. Hence, abnormal activation of proximal muscles during seated reaching affects the muscle coordination and kinematics of the distal region.Furthermore, abnormality in the activation of shoulder muscles can lead to functional impairments in the hand muscles. Dai et al. explores the relationship between the emergence of mirror movements and the quality of motor function in stroke patients. Mirror movements are involuntary movements occurring on one side of homologous muscles when unilateral voluntary movements are performed. sEMG techniques revealed mirror activities in both affected and unaffected muscles during maximal contractions. The authors used standardized net excitation (SNE) and overflow percentage (OF) to measure the strength of mirror activities. The authors found that OF in the case of unilateral contraction tasks on the affected side was significantly higher than unilateral contraction tasks on the unaffected side. The SNE appeared unrelated to the motor function of the patients but had a positive relationship with the improvement of motor function, indicating that the activation of the contralateral cortex may be involved in the potential recovery of motor function of the affected side. Hence, the high OF on the contralateral side due to the stroke resulted in highly asymmetric mirror movement.The research article by Song et al. explores the efficacy of the MScanFit program to estimate the number of motor units based on compound muscle action potential (CMAP) from the recordings of five hand muscles, i.e., first dorsal interosseous, abductor pollicis brevis, abductor digiti minimi, second lumbrical, and abductor hallucis muscles. The authors found that their motor unit number estimates were mostly consistent with previous studies. These findings suggest that the technique may be a promising tool for early diagnosis and monitoring of neuromuscular disorders.The brief research report by In summary, our Research Topic has successfully provided valuable information on the characteristics of neuromuscular disorders, particularly strokes, at the CNS level. Furthermore, it has provided effective monitoring of neuromuscular disorders. For example, MScanFit is beneficial for tracking the loss and reinnervation of motor units at different stages of stroke rehabilitation. Additionally, our topic offers therapeutic solutions for practitioners. For instance, therapy-based training involving seated reaching tasks improves motor function. Therefore, the markers identified in these studies are necessary for rehab practices. Each of these articles provides experimental, theoretical, and computational solutions for the rehabilitation of patients afflicted with neuromuscular disorders, as shown in RES, KI, and JS: writing, drafting, and editing. All authors contributed to the article and approved the submitted version.


The increasing comorbidity of alcohol use disorder (AUD) and post-traumatic stress disorder (PTSD) associated with traumatic brain injury (TBI) is a serious medical, economic, and social issue. However, the molecular toxicology and pathophysiological mechanisms of comorbid AUD and PTSD are not well understood and the identification of the comorbidity state markers is significantly challenging. This review summarizes the main characteristics of comorbidity between AUD and PTSD (AUD/PTSD) and highlights the significance of a comprehensive understanding of the molecular toxicology and pathophysiological mechanisms of AUD/PTSD, particularly following TBI, with a focus on the role of metabolomics, inflammation, neuroendocrine, signal transduction pathways, and genetic regulation. Instead of a separate disease state, a comprehensive examination of comorbid AUD and PTSD is emphasized by considering additive and synergistic interactions between the two diseases. Finally, we propose several hypotheses of molecular mechanisms for AUD/PTSD and discuss potential future research directions that may provide new insights and translational application opportunities. In recent years, the unsettling prevalence of a feature in most veterans with a history of traumatic brain injury (TBI) has been that a high proportion of post-war veterans diagnosed with a psychiatric problem so-called post-traumatic stress disorder (PTSD) also suffer from alcohol use disorder (AUD) ,2,3,4. CAUD is a chronic relapsing brain disorder characterized by an impaired ability to cease or regulate chronic and excessive alcohol misuse despite adverse social, occupational, or health consequences. The 12-month prevalence and lifetime prevalence of AUD are 13.9% and 29.1%, respectively . PatientTBI, resulting from an impact by an external mechanical force, is the leading cause of mortality and disability across all ages, both in civilian and military populations . The degNot all TBI victims suffer from PTSD, AUD, or PTSD, particularly following TBI, and they have been documented not only in military veterans but also in community patients . HoweverThis review was carried out by searching the keywords  for the topics “Molecular Toxicology” or “Pathophysiology” using MESH terms. The literature searches were performed by using the PubMed/Medline databases. Only English-language articles were chosen. We examined the titles and abstracts of articles and then obtained the full texts of potentially relevant papers. Diagnostic and Statistical Manual of Mental Disorders (DSM). Under DSM-5, anyone meeting any 2 of the 11 criteria during the same 12-month period receives a diagnosis of AUD; the severity of AUD  is determined by the number of criteria met [Alcohol (ethanol) is the most frequently used recreational agent  and playeria met ,39,40  sigAlthough some research suggests that low levels of drinking can exhibit some health benefits, AUD has been widely identified as a major risk factor for both mental and physical harm and disability ,47. PTSDAUD can cause neurochemical and behavioral changes in the brain. TBI disrupts the normal physiological state of the brain, resulting in changes in brain structure and metabolism, so patients with PTSD following TBI may have a series of alterations in brain molecular components. Despite significant advancements in understanding the molecular complexity of AUD and the dynamics of the action mechanism of alcohol misuse ,51,52, iThe following sections summarize the toxicological and pathophysiological mechanisms of PTSD, AUD, and AUD/PTSD based on the aspects of metabolomics, inflammation, neuroendocrine, signal transduction pathways, and genetic and epigenetic regulation. The normal brain’s activities and responses are regulated by the need to maintain proper homeostasis in the presence of various endogenous and exogenous challenges. Following a TBI, maintaining optimal metabolic function in the brain is critical for preventing the onset and progression of neurological and psychiatric disorders. Targeted metabolomic quantitative analysis of blood and urine specimens is a common research methodology ,72. The The metabolomic analysis of the post-mortem human brain tissues revealed that chronic excessive drinking may have a preferential effect on catecholaminergic signals . ThroughPTSD is known to be associated with metabolic dysfunction, which may be caused by changes in the composition and abundance of gut microbiota . MetabolCatalase and cytochrome P450-2E1 (CYP2E1) are the two major enzymes that are responsible for brain ethanol oxidation into acetaldehyde ,83 Tabl. In addiBrain acetaldehyde has been found to have a reinforcing role in voluntary ethanol consumption, though the mechanism remains controversial ,91,92,93Accumulating evidence suggests that glutamate neurotransmission plays a critical role in alcohol dependence. A recent study reported that glutamate is modulated by alcohol cues in the forebrain reward circuit pathway in the early stage of alcohol dependence . In the Due to the toxic effects of alcohol, especially in large amounts, the brain and other organs frequently show severe functional and structural abnormalities in AUD patients and experimental models. Additionally, the consumption of ethanol has been linked to neuronal inflammation through microglial activation, neuritis, and higher levels of monocyte chemotactic protein 1 (MCP-1) mRNA and protein expression ,63. HowePro-inflammatory cytokines, which play an important role in PTSD, are produced after TBI and participate in the upregulation of the inflammatory response. For instance, TBI primarily increases the production of pro-inflammatory cytokines, which are involved in the upregulation of inflammatory reactions and play an important role in maintaining normal brain function and repair after TBI ,101. In Over the last decade, studies have demonstrated that the dysregulation of the endogenous stress-regulation systems plays a crucial role in a variety of psychological disorders, including PTSD  and AUD It is well-established that the major neurotransmitter dopamine is involved with reward systems, motivation, euphoria, fear, fright, anxiety, and learning. According to a recent study, the physiological properties of dopaminergic neurons can predict an individual’s susceptibility to stress and anxiety . In addiIn recent years, a research direction in the study of AUD that has received increased attention is the “gut-brain axis”  and its potentially huge influence not only on physical and digestive issues but also on mental health conditions . StudiesIncreasing research evidence supports those gut-brain peptides, such as glucagon-like peptide 1 (GLP1), ghrelin, neuropeptide Y, substance P, and neurotensin, that are thought to play an important role in the molecular toxicology and pathophysiology of AUD-associated tissue damage ,124,125 Alcohol can cause glial activation, neuronal apoptosis with myelin loss, and eventually long-term neurological deficits via oxidative stress signals; abnormal cell signaling transduction is crucial in alcohol-induced brain injury . Some stPrevious research has shown that after a TBI, the brain experiences a significant change in functional connectivity and transduction, disrupting a wide range of brain circuits ,135. AnaAUD can reduce the number of synapses and cause axonal impairment in the medulla oblongata, making the brain more vulnerable to further injury. High mortality rates have been statistically linked to synaptic and axonal disruptions caused by comorbid AUD and PTSD . A growiMany studies have shown that AUD can cause extensive changes in gene expression and epigenetic regulation. In fact, long-term alcohol misuse has been linked to changes in DNA modifications and gene expression, as well as alterations in functional characteristics similar to those seen in AUD patients with abnormal behavior and decreased cognitive capacity . DiffereAccording to the study of alcohol-related gene variation and brain function change, the severity of AUD-related consequences seems to be related to the altered responses of the frontal lobe of the brain to ethanol signals; the genetic variation of dissimilar neural plasticity and signal pathways is the main reason for this different response . Genome-Differential gene expression and epigenetic changes are known to be controlled by a complex set of regulatory systems. Negative changes in genomic DNA and epigenetic regulation can cause a variety of CNS disabilities. A genome-wide association study revealed that PTSD is found to be strongly linked with a single nucleotide polymorphism (SNP) in the coding region of CRP . FurtherCYP2E1 is a major ethanol-inducible enzyme and is involved in alcohol-induced oxidative tissue injury . ChronicNumerous researchers recognize the theory that alcohol exposure can regulate epigenetics as a factor in the development and progression of AUD . Meta-anLong noncoding RNAs (lncRNAs) play an important role in brain development and synaptic plasticity. The BDNF-AS lncRNA regulates synaptic plasticity in the amygdala of AUD patients who begin drinking in adolescence via epigenetic reprogramming . EthanolAt the transcriptional level, ethanol inhibits the upregulation of a subset of immediate–early genes encoding for transcription factors, including Atf3, c-Fos, FosB, Egr1, Egr3, and Npas4, but does not affect the upregulation of others   Table 3Table 3. Emerging from the review, we summarize some interesting data in Based on the above discussion about the molecular toxicology and pathophysiological mechanisms that may lead to AUD/PTSD, we further propose the following hypotheses for the onset and progression of AUD/PTSD.Due to the prolonged stressed state resulting from TBI, normal physiological regulating mechanisms in the brain may be disrupted and metabolic dysfunction with neuroendocrine abnormalities may appear in AUD/PTSD. Furthermore, an aberrant buildup of several metabolic intermediates may raise the chance of developing comorbid AUD and PTSD. Consequently, metabolic dysfunction is likely to play an important role in the development and progression of AUD/PTSD.Biochemical changes in the body, as well as the activation of microglia and astrocytes, all contribute to the major inflammatory responses in the brain of AUD/PTSD. Consequently, a slew of pro-inflammatory cytokines and chemokines are activated and released into the extracellular space. This chronic inflammatory status causes structural and functional changes in the brain. In this regard, pro-inflammatory cytokine levels could be used as biomarkers for quantifying AUD/PTSD at an early stage of progression. As a result, an exaggerated inflammatory response may be a critical factor in the onset and duration of AUD/PTSD.Neuroendocrine homeostasis is the foundation of the body’s normal functions. Under stimulation factors such as alcohol misuse, TBI, and PTSD following TBI, the neuroendocrine homeostasis, particularly in the nervous system, is disrupted and the levels of various neurotransmitters and their receptors are altered. The stimulation factors continue to exist and result in the continuation and development of AUD/PTSD, leading to the formation of a vicious circle. Neuroendocrine changes are not only an important cause of the onset of AUD/PTSD but also a key factor in its persistence through the formation of a vicious cycle. The investigation of neuroendocrine dysfunction in vivo is critical in determining the persistent states of AUD, PTD, and AUD/PTSD. AUD and PTSD can cause changes in brain homeostasis, which can alter gene expression and/or epigenetic regulation. Patients and experimental rodents with altered expression of various genes governing the sensitivity within cerebral neurons may eventually exhibit a variety of abnormal behaviors and decreased cognitive functions. However, the conditions and mechanisms regulating gene expression in vivo remain unknown; additional research is needed to fully understand their precise mechanisms. Thus, genetic and epigenetic changes may play a crucial role in the final development of AUD/PTSD, which is also an important reason why this comorbid disease is difficult to treat.Based on our summary of the various mechanisms, we are currently considering the following scenario. Although there are various hypotheses for the molecular mechanisms of comorbid AUD and PTSD, none of them can perfectly explain its pathogenesis. Therefore, we consider that AUD/PTSD may be the result of the combined action of multiple pathogenic factors. The destruction of anatomical structure and the changes in neurotransmitters and their receptors may lead to alterations in metabolism, neuroendocrine function, pro-inflammatory factors, and others in the body. At the same time, the abnormality of the signal transmission system and neural function in the body caused by mental stress may lead to genetic and epigenetic changes. Finally, the aforementioned factors caused by the psychiatric disorder PTSD and/or AUD interact to cause AUD and PTSD comorbidity. Thus, comorbid AUD/PTSD results from the combination of several complex risk factors . Because the causal relationship and mutual influence among AUD, PTSD, and AUD/PTSD are complex, many theories such as the self-medication hypothesis have been proposed to understand their relationship . For insA lack of understanding of the disease’s molecular pathogenesis results in improper prevention and intervention in current treatments. This is also true in the case of comorbid AUD/PTSD. Therefore, we suggest that future studies should focus on the following aspects. (1) Molecular mechanisms of the enzymes such as catalase, CYP2E1, and ALDH2 are involved in the cerebral ethanol metabolism and oxidative stress, leading to brain injury before and during AUD/PTSD. (2) The HPA axis is principally in charge of stress neuroendocrine control and the recovery of post-stress homeostasis following stressful events. Studies on psychiatric patients diagnosed with more than a single mental disorder or on substance users with psychiatric disorders have shown that the co-occurrence of psychiatric disorders might be an additional correlate of HPA-axis dysregulation ,171. RegAlthough many molecular mechanisms that may lead to AUD/PTSD comorbidity have been studied, the toxicological and pathophysiological mechanisms of AUD/PTSD remain largely unknown. Our review focuses on the molecular toxicology and pathophysiology of AUD and PTSD comorbidity, with significant additive or synergistic effects on the individual symptoms, compared with those suffering from each disease alone. We also emphasize that the patients with AUD/PTSD should be thoroughly examined and that the pathogenesis should be discussed in multiple aspects instead of under separate testing and consideration. We specifically discussed the molecular mechanisms of AUD, PTSD, and AUD/PTSD from metabolomics, including changes in molecular components in the brain, signal transduction pathways, gene expression, and epigenetics, and suggested possible hypotheses for the development of AUD/PTSD. This review focuses on certain aspects of molecular toxicology and pathophysiology mechanisms. Further studies on the causal relationship and mutual influence among AUD, PTSD, and AUD/PTSD are encouraged.


This mini-review was designed primarily to explore the commonly employed in organello analytical assays for the assessment of mitochondrial physiology and its dysfunction, with a particular focus on neuroscience research. The authors briefly discuss the methodologies for biochemical isolation of mitochondria, their quality assessment, and cryopreservation. Further, the review attempts to accumulate the key biochemical protocols for in organello assessment of a multitude of mitochondrial functions critical for neurophysiology, including assays for bioenergetic activity, calcium and redox homeostasis, and mitochondrial protein translation. The purpose of this review is not to examine each and every method or study related to the functional assessment of isolated brain mitochondria, but rather to assemble the commonly used protocols of in organello mitochondrial research in a single publication. The hope is that this review will provide a suitable platform aiding neuroscientists to choose and apply the required protocols and tools to address their particular mechanistic, diagnostic, or therapeutic question dealing within the confines of the research area of mitochondrial patho-physiology in the neuronal perspective.Mitochondria regulate multiple aspects of neuronal development, physiology, plasticity, and pathology through their regulatory roles in bioenergetic, calcium, redox, and cell survival/death signalling. While several reviews have addressed these different aspects, a comprehensive discussion focussing on the relevance of isolated brain mitochondria and their utilities in neuroscience research has been lacking. This is relevant because the employment of isolated mitochondria rather than their  Mitochondria, the cellular power houses, hence are critical mediators of neuronal physiology, fulfilling the high-energy demands of neurons. In addition, mitochondria regulate calcium and redox signalling, and cellular death through regulation of the apoptotic and necrotic signals -radiolabelled cysteine and methionine, followed by either gel autoradiography or radioactivity counting [in organello mitochondrial protein translation. It will be interesting to follow the development of alternative non-radioactive methods for the evaluation of mitochondrial protein synthesis, such as those based on puromycin [Local counting , 170-173uromycin , 169, anuromycin . Intriguuromycin .6e.g., synaptic versus non-synaptic mitochondria; [e.g., embryonic versus postnatal, or young versus aged [e.g., in Alzheimer’s disease [e.g., bis(7)-tarcine [Proteomic characterization is the most obvious and most widely employed application of isolated mitochondria, both in terms of the global proteomic characterization of mitochondrial proteins and their post-translational modifications. Both gel-based and gel-free comparative mitochondrial proteomics methodologies have been widely-used to evaluate the quantitative and qualitative differences in mitochondrial proteome as a function of subcellular localization , develosus aged ), diseas disease ; and mul disease ) as well-tarcine ; and sim-tarcine ). Since -tarcine , 181-1907in organello assays is essential in order to evaluate the actual inherent mitochondrial properties, independent of the differences in the mitochondrial content of different preparations. There are several ways to do this. The simplest of these are normalizing mitochondrial protein content using the established protein quantitation assays such as Bradford or bicinchoninic acid (BCA) methods. However, this method is not specific for mitochondria  and hence may be erroneous. A better normalization control is the activity of CS [Normalization for ty of CS , 191. Inex vivo mitochondrial preparations in the characterization of the physiology and pathophysiology of neuronal mitochondria, it is clear that the use of these preparations has been under-utilized in neurobiology research.While studies have provided some evidences for the utility of In particular, research on the synaptic mitochondrial pool is lagging behind, in spite of the knowledge that synapses are the major sites of interneuronal communication and substrates for almost all higher-order functions and their dysfunction is a common and primary pathogenic event in brain pathologies across the whole development span, from early-life insults to ageing-related neurodegeneration. Given the critical roles of synaptic mitochondria in neurotransmission, it is surprising that only a few studies have been directed against evaluating their functions specifically. In this regard, biochemically isolated synaptic mitochondria seem to be the most important step forward to evaluate their function (and dysfunction) in relation to synaptic physiology and brain functions.1). However, in doing so, the hope is that it will help neuroscientists to identify and appreciate the utilities of these biochemical fractions and encourage them to employ their expertise in understanding the finer complexities of mitochondrial (and synaptic) pathology in healthy and diseased brain states. We also hope that through this review, researchers will be motivated to identify the gaps and device novel assays to evaluate mitochondrial (patho)physiology. One can only hope for an escalation in the employment of ex vivo mitochondria from cellular and animal models and human subjects to further delineate the contribution of mitochondria to brain function and dysfunction, and to evaluate the efficiency of potential therapeutic agents/strategies in prevention and amelioration of multiple neuropathologies.The purpose of the review is to solely summarize the methodologies and protocols of mitochondrial research, specifically using isolated brain mitochondrial samples (Table


Sponges are early-derived and simple metazoans that have significant functions in marine ecosystems, including sediment stabilization, nutrient cycling, and the provision of habitats for numerous species . The incbona fide or opportunistic pathogens for sponges as they behave similarly with other animals, including corals  microbial taxa that are found in all individuals of a species is regarded as the core microbiota . The ecoSuberites domuncula might recognize fungi via the β-glucans on their cell walls, but the specific effect of the cytokine produced by the activation of this signaling pathway has not been described  in the Indian Ocean were affected by an unknown disease that was most probably caused by a filamentous fungus. Later, mortality between 70 and 95% of the affected individuals was recorded in commercial sponges in the Florida Keys (1939), Bahamas (1938–1939), Cuba (1939), and British Honduras (1939)  aff. Biru sponges colonizing Maldivian ecosystems  . Again, s (1939) . In 2015osystems . The indAspergillus sydowii) may infect corals, there is, to the best of our knowledge, only one recent study demonstrating that a fungus (Aspergillus tubingensis) can cause a fatal infection in the marine sponge Chondrosia reniformis  and facilitate long-term simulations, allowing for multi-generational studies. This approach would allow the identification of fungal taxa that could be harmful to marine sponges at different temporal scales. Moreover, it may also provide useful insights into fungal signatures, whether in the form of community structure or potential functionality, that could serve as predictors of sponge health within marine ecosystems.To advance our understanding of emerging fungal pathogens, it is crucial to implement novel experimental approaches . Until rin vitro  and 3D cin vitro , which sIn recent years, the potential use of microbial transplants and probiotics for treating a wide variety of human and animal disorders have emerged as novel paradigms . MicrobiPseudoalteromonas sp., Holomonas taeanensis, and Cobetia marina-related species, isolated from the coral Pocillopora damicornis, partially mitigated coral-bleaching and significantly improved the coral resistance to water warming. In addition, inoculation with this consortium controlled the development of the temperature-dependent pathogen Vibrio coralliilyticus . This output reflects only the author’s view, and the Research Executive Agency (REA) cannot be held responsible for any use that may be made of the information contained therein. YP-L received postdoctoral fellowships from UNAM-DGAPA and from CONACyT (CVU: 697462). RB-G received a Sabbatical fellowship (CVU: 389616) from the National Council of Science and Technology (CONACyT), Mexico.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Cardiomyopathy develops through an interaction of genetic and environmental factors. The clinical manifestations of both dilated cardiomyopathy and hypertrophic cardiomyopathy are diverse, but genetic testing defines the causative genes in about half of cases and can predict clinical prognosis. It has become clear that cardiomyopathy is caused not only by single rare variants but also by combinations of multiple common variants, and genome-wide genetic research is important for accurate disease risk assessment. Single-cell analysis research aimed at understanding the pathophysiology of cardiomyopathy is progressing rapidly, and it is expected that genomic analysis and single-cell molecular profiling will be combined to contribute to more detailed stratification of cardiomyopathy. Thirty years have passed since it became clear that the cause of cardiomyopathy is genetic mutations, and precision and genomic medicine based on an understanding of the pathogenesis of individual patients. In this paper, we will summarize the current findings in advancing genomic medicine for cardiomyopathy, and discuss future perspectives for the development of optimal treatment methods based on molecular pathology and the establishment of more precise stratification integrated with artificial intelligence.TTN gene), Lamin A/C (encoded by the LMNA gene), and Desmoplakin (encoded by the DSP gene). DCM develops when mutations (mainly rare single-nucleotide mutations) occur in these gene regions and abnormal proteins are produced.Dilated cardiomyopathy (DCM), characterized by diffuse contractile dysfunction of the left ventricle and dilatation of the left ventricular lumen in the absence of abnormal stress conditions, such as hypertension, valvular disease, and coronary artery disease, and is a progressive disease with poor prognosis with repeated exacerbations of heart failure symptoms . It has TTN gene, the most common DCM causative mutations, can cause alcoholic cardiomyopathy . A genome cohort study revealed that DCM cases with pathogenic or likely pathogenic mutations in known causative genes show a poorer clinical prognosis compared to cases without these mutations .TTN truncation mutations, which account for 12%–25% of DCM, were a favorable prognosis group with reverse left ventricular remodeling (restoration of systolic function) in response to medical therapy, whereas LMNA gene mutations, which account for 4%–10% of DCM, were a poor prognosis group without reverse remodeling  is a group of disorders characterized by hypertrophy of the left or right ventricular myocardium and left ventricular diastolic dysfunction due to cardiac hypertrophy. Mutations in sarcomere genes are considered to be the main cause of HCM , and theThe availability of genetic testing has enabled genetic screening of cardiomyopathy families to identify asymptomatic family members harboring mutations that can cause cardiomyopathy. Lorenzini et al. conducted a 15-year clinical follow-up of cardiomyopathy mutation-positive family members who did not initially display the HCM phenotype, and found that approximately 50% of these family members developed HCM . In partGenetic analysis also increases the possibility of reaching an accurate diagnosis for diseases showing phenotypes similar to HCM, such as Fabry disease and transthyretin cardiac amyloidosis; therefore, the importance of genetic testing in routine clinical practice of cardiomyopathy is clearly increasing. The clinical guideline regarding the interpretation and actionability of genetic testing in cardiomyopathy has also been published by the European Society of Cardiology . Furtherβ signaling and downregulation of genes related to mitochondrial metabolism are hallmarks of cardiomyocytes of patients with DCM , showed high expression of intercellular adhesion-related genes, and group 4, which showed severe cardiac dysfunction, showed high expression of DNA replication-related genes.Cardiomyopathy develops through an interaction of genetic and environmental factors. Intracellular phenotypes should appear as endophenotypes at the time of disease onset, and identification of such phenotypes will lead to disease stratification and treatment development . Recent with DCM , 32 and with DCM . Wang etwith DCM . Ko et awith DCM . Verdonswith DCM . The expLMNA mutant cardiomyopathy, whereas IL6 signaling was activated in TTN mutant cardiomyopathy. Ameen et al. analyzed the human fetal heart by single-cell ATAC-seq  to profile cell-type-specific epigenomic changes during development , DCM, and HCM, and revealed CHD-specific cell states in cardiomyocytes, which showed characteristics of insulin resistance and increased expression of genes associated with FOXO signaling . They alafter MI , 43. Kupafter MI . They idIn this article, we reviewed recent papers that clarified the genetic characteristics of DCM and HCM, and discussed the direction for the development of precision and genomic medicine in cardiomyopathy. Furthermore, we introduced an attempt to understand the molecular pathogenesis of DCM and HCM using single-cell analysis technology, and to promote disease stratification based on the mechanisms. Here, we discuss the challenges in constructing precision medicine based on genetic testing, and the future perspectives of pathophysiological analysis based on the development of single-cell omics analysis technology .To promote precision medicine based on genetic testing, it is first necessary to generalize the method of mutation analysis and establish a standard genetic testing pipeline. It is also important to develop a method for accurately evaluating the pathogenicity of identified variants. Determining pathogenicity according to the guidelines of American College of Medical Genetics and Genomics  is commoSingle-cell omics analysis has made great progress in understanding the pathogenesis of cardiomyopathy, but at this point, the molecular profiling of cell types has only been completed, and it is still far from being connected to medical applications. First, it is important to identify the molecular mechanisms associated with processes that may lead to treatment decision making, and to establish a simple disease stratification method based on these mechanisms. By combining with spatial analysis and comprehensive functional analysis, we need to connect our understanding of RNA expression levels with transcriptional regulation, signaling pathways, cell-cell interactions, and therapeutic interventions. It is also important to promote genome analysis, single cell omics analysis, and medical artificial intelligence in an integrated manner, and to establish a method for integrated analysis of the data obtained by these modalities . FurtherPaper writing: SN and MO; Supervision: MO. All authors contributed to the article and approved the submitted version.


Papaver somniferum. The use of opioids and their negative health consequences among people who use drugs have been studied. Today, opioids are still the most commonly used and effective analgesic treatments for severe pain, but their use and abuse causes detrimental side effects for health, including addiction, thus impacting the user’s quality of life and causing overdose. The mesocorticolimbic dopaminergic circuitry represents the brain circuit mediating both natural rewards and the rewarding aspects of nearly all drugs of abuse, including opioids. Hence, understanding how opioids affect the function of dopaminergic circuitry may be useful for better knowledge of the process and to develop effective therapeutic strategies in addiction. The aim of this review was to summarize the main features of opioids and opioid receptors and focus on the molecular and upcoming epigenetic mechanisms leading to opioid addiction. Since synthetic opioids can be effective for pain management, their ability to induce addiction in athletes, with the risk of incurring doping, is also discussed.Opioids are substances derived from opium . In its raw state, opium is a gummy latex extracted from  Papaver somniferum have been used recreationally as a euphorigenic or medicinally as analgesic for the treatment of pain . A. A104]. + dependent hyperpolarization or a Cav2.1 mediated depolarization in different VTA neurons . In fact, murine model of human OPRM1 A118G (A112G in mouse) displayed a weaker synaptic modulation of GABA and glutamate release onto VTA DA neurons [COMT is a key regulator of catecholamines  methylation that influence their concentration in the pain perception pathways. Rs4680  and rs4818 (C > G) are the main polymorphisms in COMT gene, with a prevalence of variants of 50% and 40% in Caucasians, 28% and 17% in African-Americans, 44% and 31% in Asians, respectively [OPRM1 polymorphism, there is a paucity of evidence linking COMT to interindividual variation in opioid sensitivity  is the study and clinical application of the role of genetic variation on drug response. The Clinical Pharmacogenetics Implementation Consortium (CPIC), in recent times, published a clinical practice guideline on the use of pharmacogenetic information for opioid therapy for pain control. This guideline includes recommendations for how rocodone ,178. CYPt review ,180. CYPe (COMT) ,180. Wit neurons , thus poTherefore, scientific evidence suggests that individual variabilities can be attributed to single nucleotide polymorphisms in genes involved in opioid pharmacodynamics and pharmacokinetics. Knowledge of these genetic factors through pharmacogenetic testing can help clinicians to prescribe a more adequate opioid that can offer patients with maximal clinical benefit and minimal risk of adverse effects.As research unknots the various genetics, biochemical and receptor interaction differences of opioids in humans, it is expected that certainly obtainable, cost-effective testing will become available to aid clinicians in choosing an optimal opioid analgesic for an individual patient ,181. PGxStudies on epigenetics and pain remain largely preclinical and investigate the theoretical ability of epigenetics to alter the nociceptive pathways both in the periphery and centrally . MechaniH19, BC1, MIAT1 and MALAT1 lncRNA were investigated in rats in specific brain areas  [The main limitations of morphine use in pain control are analgesic tolerance and dependence. The effects of repeated injection of morphine on the expression of triatum) . AlteratCircNf1 resulted in being highly expressed in the dorsal horns of morphine-treated rats and, acting as a sponge for miRNA665 and miRNA330-3p, enhances the expression of the CXC motif chemokine ligand 12 (CXCL12), a homeostatic chemokine essential not only in the modulation of morphine analgesic effects in chronic pain but also in neurodevelopmental process, maturation of the CNS, neuronal toxicity and transmission, neuroglial interactions and nerve regeneration [Lastly, recent findings revealed the involvement of circRNA to cytokine pathway in order to mediate morphine analgesic tolerance . In partneration ,187.The abuse of pharmacological active substance which improve athletic performance may alter the expression of specific genes involved in muscle and bone metabolism by epigenetic mechanisms, such as DNA methylation, histone tails modifications and ncRNA production . The preOpioids are prescribed in a variety of settings for treatment of both acute and chronic pain. However, studies demonstrating benefits of long-term use of opioids to manage chronic, non-cancer pain are lacking. Moreover, the evidence clearly demonstrates that long-term use of opioids is associated with an increased risk of OUD and overdose, as well as a series of other adverse outcomes  ,162,169.Drug addiction is the consequence of long-lasting adaptions in brain reward and non-reward neuronal networks; hence, understanding how opioids affect neuroplasticity in mesocorticolimbic dopaminergic circuitry may be enable better knowledge of the process and the development of effective therapeutic strategies. Recently, most studies have been focused on the epigenetics signature of tissues and cells, which consists in a large plethora of chemical modifications in the DNA or histone proteins and functionally adapts the chromatin status and, in turn, gene expression to lifestyle and environment. Following opioid exposure, the epigenetic signature changes to different extents in the brain and periphery. However, the primarily maintenance medications of opioid addiction, i.e., methadone, also changes the epigenetic signature in terms of the DNA methylation status . Hence, It is known that, generally, an opioid such as tramadol morphine or codeine  is used widely in sport to treat pain and inflammation associated with injury , but intA variety of effects can occur after a person takes opioids, ranging from pleasure to nausea and vomiting, severe allergic reactions (anaphylaxis) and overdose, in which the breathing and heartbeat slow or even stop. Medical doctors should be encouraged to keep their knowledge current about evidence-based practices for the use of opioid analgesics to manage pain, as well as specific steps to prevent and manage opioid overdose. However, the tailoring of medicine—on the road to personalized medicine—considers the molecular and genetic mapping of the individual, even if many factors still impede the smooth application of personalized medicine and represent challenges or limitations in its realization.


Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter species) pathogens, which include four Gram-negative bacterial species. In these bacteria, active extrusion of antimicrobial compounds out of the cell by means of ‘molecular guns’ known as efflux pumps is a main determinant of MDR phenotypes. The resistance-nodulation-cell division (RND) superfamily of efflux pumps connecting the inner and outer membrane in Gram-negative bacteria is crucial to the onset of MDR and virulence, as well as biofilm formation. Thus, understanding the molecular basis of the interaction of antibiotics and inhibitors with these pumps is key to the design of more effective therapeutics. With the aim to contribute to this challenge, and complement and inspire experimental research, in silico studies on RND efflux pumps have flourished in recent decades. Here, we review a selection of such investigations addressing the main determinants behind the polyspecificity of these pumps, the mechanisms of substrate recognition, transport and inhibition, as well as the relevance of their assembly for proper functioning, and the role of protein–lipid interactions. The journey will end with a perspective on the role of computer simulations in addressing the challenges posed by these beautifully complex machineries and in supporting the fight against the spread of MDR bacteria.Bacterial resistance to antibiotics has been long recognized as a priority to address for human health. Among all micro-organisms, the so-called multi-drug resistant (MDR) bacteria, which are resistant to most, if not all drugs in our current arsenal, are particularly worrisome. The World Health Organization has prioritized the ESKAPE ( Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter species) pathogens prioritized by the World Health Organization . X. X148]. Owing to their functional interaction with the RND transporters, MFPs have been considered a good target for developing inhibitors . In factE. coli  was proposed as the lead compound. Seyedhosseini-Ghaheh et al. [Several groups employed a et al.  performeh et al.  identifiDespite the key role of protein–protein and protein–membrane interactions in mediating polyspecificity and transport by RND pumps , unveiliet al. performed the first MD simulations of an intact MexAB-OprM pump in a P. aeruginosa bacterial membrane envelope model [In 2017, López pe model . By combpe model . Moreoveet al. explored its conformational dynamics by combining mutagenesis and functional experiments with MD-based free-energy calculations [Two years later, in an effort to shed some light on the role of AcrA in the assembly of AcrAB-TolC, Hazel ulations . The estulations .E. coli to novobiocin and erythromycin antibiotics. This demonstrated the importance of AcrA conformational flexibility for the assembly of AcrAB-TolC and confirmed that modulation of its dynamics through pharmacological intervention is a possible approach to develop new antibiotics.Site-directed mutagenesis experiments, introduced at an interdomain interface to shift the dynamic equilibrium in favour of the incompetent one, disrupted pump assembly and function and resensitized et al. [P. aeruginosa due to the G72S substitution in MexA [More recently, Cacciotto et al.  performe in MexA . The aut in MexA ; and  and increased antibiotic susceptibility in A. baumannii [Zang mannii . CG MD sin silico studies have provided valuable clues to several aspects of RND efflux pumps. These include insights into the molecular determinants for their polyspecificity, the mechanisms for substrate transport and those behind inhibition, the assembly of tripartite assembly and the relevance of lipid–protein interactions. These studies have not only supported and rationalized experimental findings, but have also provided original explanations and inspired new research. In the following, we summarize a few methodological developments that could increase the impact of in silico investigations on RND efflux pumps and suggest possible future directions for computational studies.In the last few decades, First, while all-atom MD simulations have yielded numerous insights into the molecular aspects of the components of RND machineries, in bacteria these pumps perform their function within an extremely complex environment constituted by the real membrane. The interactions between different components of the tripartite system, and of these proteins with other membrane and periplasmic proteins and with the molecules within the inner and outer membranes could have synergic effects modulating the efflux of compounds outside the cell . It has in vivo counterparts. The importance of such computational studies is reinforced by the limitations of experiments and theoretical models in detecting and modelling the intrinsic complexity of real membranes [To provide an example of how these interactions can be crucial, one can think about the still unclear allosteric conformational regulation occurring between the MFP and IMP proteins along the functional cycling of the pump , or to tembranes . SummariAlthough the polyspecificity of the efflux pumps appears to hamper the development of new effective antimicrobials, this phenomenon could also represent an opportunity in drug discovery, as in the case of repurposing approaches, that could benefit from the ability of the transporters to bind multiple ligands, including already approved drugs . Also inAlthough the idea of using EPIs to improve the efficacy of existing antibiotics holds a great deal of promise, pursuing non-toxic, safe and efficient molecules having such activity has faced so far many issues, and no EPI has been approved to date for treatment in MDR bacteria . A majora priori from the chemical formula and/or the three-dimensional structure. Among the descriptors mostly used to build QSAR models are various types of topological fingerprints, such as the Extended-Connectivity Fingerprints . Graph convolutional networks (GCNs) represent an interesting method to extract several parameters (descriptors) from molecular structures. A paradigmatic example of application of these methodologies in the context of antimicrobials led to the identification of the compound halicin  as a broad-spectrum antibiotic, showing activity against drug-resistant strains of different bacteria [Concerning the role of machine learning methods, several studies have already reported their use as a tool to unveil the relationship between the molecular properties of compounds and their propensity to be a substrate and/or inhibitor of efflux systems together with their ability to cross bacterial membranes. The identification of quantitative relationship between structure and properties/activity (QSPR/QSAR) of (candidate) drugs is very useful to filter out thousands of inactive compounds from virtual databases. Usually, machine (deep) learning algorithms are fed with a set of molecular descriptors generated bacteria . Along tTo summarize, tripartite efflux pumps underpin several different mechanisms of resistance, virulence and biofilm formation. As such, they are crucial players of antimicrobial resistance, and an important challenge in the development of antimicrobial compounds concerns the identification of active drugs avoiding efflux systems, and/or inhibitors of the latter systems. In the recent years, significant progresses have been made in understanding the physicochemical properties making a drug susceptible to being transported by efflux systems and the specific regions of these machineries involved in transport. However, due to the extreme complexity of these proteins and despite its importance for the design of more effective drugs, a complete understanding of the mechanisms underpinning substrate efflux is still missing. In this context, computational studies have demonstrated to be of great assistance not only to aid the development of new drugs with specific (desired) properties, but also in understanding their fate with the efflux pump. Thanks to the continuous development of more accurate models, community-based tools for the automation of simulations, and increased computing power, we are now able to simulate these systems on a larger time scale and in more realistic and complex environments, enabling researchers to capture key structural and dynamic aspects of these beautiful and so daunting machineries.


The development of the Gonadotropin-releasing hormone (GnRH)-neuron system is a fascinating and complex process that plays a crucial role in the regulation of reproductive function. These hypothalamic neuroendocrine neurons control pubertal onset and fertility in most vertebrates through the strictly regulated release of the GnRH peptide, consisting of pulses and surges. Interestingly, GnRH-neurons developmentally originate outside the brain, from stem cells located in the nasal placode. During embryogenesis, the GnRH-neurons migrate along the axons of the terminal nerve to arrive at the hypothalamus, and this migration also relies on the correct formation of olfactory ensheathing cells (OECs), an olfactory system-specific glial population that migrate with the GnRH-neurons .After the migration of the GnRH-neurons and in early postnatal life, they undergo alterations in their activity, biosynthetic profile, and connectivity, including axon elongation to the median eminence. The development of the GnRH neuron system is a highly dynamic process that involves multiple stages and a complex interplay of genetic, epigenetic, environmental, and hormonal factors. Unraveling the contributions and interactions of GnRH-neurons with other cells and signaling molecules, as well as dissecting the precise timing and sequence of events during GnRH neuron development is a major challenge in understanding the physiopathological aspects of GnRH neuron system development.Despite these challenges, ongoing research efforts, including the use of advanced technologies and improved animal models, are gradually improving our understanding of the physiopathological aspects of GnRH neuron system development. The integration of multiple disciplines, such as developmental biology, genetics, neurobiology, and endocrinology, is crucial for making progress in this field. In this editorial, we will explore the physiological and pathological aspects of GnRH neuron system development, shedding light on the intricacies of this fascinating field. Join us as we delve into the latest research and gain a deeper understanding of the mechanisms that underlie this critical system.Xie et al. Understanding the influence of kisspeptin on the reproductive axis and related mechanisms could have important implications for disease diagnosis and treatment.An editorial on GnRH neuron physiology would be incomplete without acknowledging the pivotal role of kisspeptin research in unraveling the intricate mechanisms underlying the regulation of reproductive processes in mammals, beautifully reviewed from Shen et al. employ optogenetics to selectively stimulate Kisspeptin neurons in the arcuate nucleus (ARH) and anteroventral periventricular nucleus (AVPV) in order to investigate their role in the luteinizing hormone (LH) surge. The authors also examine the effects of natural steroid hormone milieu and the release of glutamatergic neurotransmitters on LH secretion. This study adds new insights into the mechanisms underlying the LH surge and may potentially contribute to improving reproductive outcomes in mammals. Indeed, altered responses to kisspeptinergic neurotransmitter systems may directly impact reproductive physiology. The study from Bhattarai et al., by examining the response of various neurotransmitters and neurosteroids regulating GnRH neuronal activities between letrozole-induced PCOS and normal mice, provides new insights into the possible neuroendocrine disruptions in PCOS at the GnRH level. The authors conclude that PCOS may directly affect the neurotransmitter system regulating GnRH activity at the hypothalamic level, thus opening up new avenues for therapeutic targets for PCOS.To this aim, the study from DrButruille et al. investigate the role of glial fibrillary acidic protein (GFAP)-expressing tanycytes in the male reproductive function and their impact on the GnRH system within the hypothalamus. The findings of this study support the view that GFAP-positive tanycytes play a critical role in the regulation of male reproductive function. The depletion of these cells alters hormone levels, testicular weight, and sexual behavior, in addition to causing significant structural alterations that may hamper the tanycyte-to-GnRH-neuron communication processes controlling reproduction. The study provides new insights into the functional and morphological interactions between GFAP-expressing tanycytes and GnRH axon terminals in the hypothalamus, highlighting their central role in maintaining the neuroendocrine system.The microenvironment of GnRH neurons participates in the modulation of their activity, particularly in the synthesis and secretion of GnRH. Tanycytes are able to form self-renewing neurospheres and produce primarily astrocytes, and their removal may modify the glial microenvironment around the GnRH neurons and fibers. Here, Chung et al. emphasize the need to focus on how GnRH neurons initiate and maintain their identity during prenatal and postnatal periods, addressing two questions: can a disease state such as CHH lead to the postnatal loss of fully differentiated GnRH neurons, and can the postnatal GnRH neuronal loss be reversed by beneficial cues?Overall, understanding the maintenance of the postnatal GnRH system for the treatment of fertility issues in humans and other animals could be of major importance since disruption of processes responsible for the initiation and maintenance of GnRH neuronal identity can lead to reproductive disruption similar to the congenital hypogonadotropic hypogonadism (CHH) phenotype. In their review article, In conclusion, this Research Topic summarizes and discusses the most recent findings about the intricate biological networks underlying GnRH neuron system, with a specific highlight on kisspeptin neurons and tanycytes that are increasingly emerging as fundamental for the correct functioning of GnRH neurons, and without neglecting a glance towards novel therapeutic targets of GnRH neuron-related disorders.RO: Writing – review & editing. KC: Writing – original draft.


Traditional echocardiographic imaging mainly identifies cardiac diseases when symptoms are clear. This makes the innovation of new methods for the prediction and early diagnosis of cardiac diseases inevitable, especially when diastolic function evaluation is needed. Generally, non-invasive assessment of heart function is currently attracting great attention away from sophisticated techniques. With the development of color M-mode echocardiography (CMME), the derived intraventricular pressure differences (IVPDs) and intraventricular pressure gradients (IVPGs) created a breakthrough in the non-invasive assessment of diastolic function in animal models as well as in some companion animals and human studies. They are beneficial for continuous data acquisition without invasive interference, and their results showed good relatedness with the measurements acquired from the catheterization technique.The assessment of diastolic function has received great interest in order to comprehend its crucial role in the pathophysiology of heart failure and for the early identification of cardiac events. Silent changes in the intraventricular flow (IVF) dynamics occur before the deterioration of the cardiac wall, although they cannot be detected using conventional echocardiography. Collective information on left ventricular (LV) pressures throughout the cardiac cycle has great value when dealing with patients with altered hemodynamics. Accurate pressure measurement inside the ventricle can be obtained by invasive methods to determine the LV diastolic pressures, which reflect the myocardial relaxation and compliance. However, catheterization is only feasible in the laboratory setting and is not suitable for clinical use due to its disadvantages. In contrast, echocardiography is simple, safe, and accessible. Color M-mode echocardiography (CMME) is an advanced cardiac evaluation technique that can measure the intraventricular pressure differences (IVPDs) and intraventricular pressure gradients (IVPGs) based on the Doppler shift of the IVF. Recently, the assessment of IVPD and IVPG has gained growing interest in the cardiovascular literature in both animal and human studies as a non-invasive method for the early diagnosis of cardiac dysfunctions, especially diastolic ones. The usability of IVPD and IVPG has been reported in various surgically induced heart failure or pharmacologically altered cardiac functions in rats, dogs, cats, and goats. This report aims to give an overview of the current studies of CMME-derived IVPD and IVPG in animal studies and its feasibility for clinical application in veterinary practice and to provide the prospects of the technique’s ability to improve our understanding. Heart failure is one of the most common causes of death globally; these cases include both diastolic and systolic dysfunctions. In humans and companion animals, heart failure is the second leading cause of death after cancer ,2. The dCurrently, it is believed that measuring diastolic function is crucial for a thorough understanding and early identification of heart diseases since diastolic dysfunction (DD) precedes systolic dysfunction in many cardiac disorders ,7. DiffeDespite numerous methods that have been established to determine diastolic performance, the evaluation of diastolic parameters by catheterization remains the gold standard method which can efficiently measure compliance, relaxation of the myocardium, and filling pressure. However, its invasiveness and the requirement of preoperative measures limit its clinical applicability ,12. LikeRecently, with the development of color M-mode echocardiography (CMME), the derived intraventricular pressure differences (IVPDs) and intraventricular pressure gradients (IVPGs) have created a breakthrough in the non-invasive assessment of diastolic function in animal models as well as in some companion animals and human studies. A literature search was conducted in English through PubMed databases and the publications included in this review are up to January 2023. Selected articles were chosen based on the presence of animal studies having either experimental, preclinical, or clinical trials using the following keywords: color M-mode echocardiography, intraventricular pressure gradients, IVPG, intraventricular pressure difference, IVPD, diastolic function.Generally, it is widely believed that silent changes in the IVF dynamics occur earlier than deteriorations of the cardiac wall; however, IVF changes cannot be detected by conventional echocardiography. The development of Doppler echocardiography has allowed non-invasive measurement of the intracardiac flow. In the normal heart, there is an IVF dynamic including pressure differences. The IVPD is generally defined as the pressure difference that is generated in early diastole between distinct segments within the LV when the apical pressure in the LV drops below the basal pressure . PressurAssessment of IVPD was first uncovered in 1979 when Ling et al.  reportedThe CMME-derived IVPD measurements have started to be used as a non-invasive metric for cardiac function evaluation because of the rapid advancement of echocardiographic technology and the introduction of CMME. Since the IVPD has been approximated from CMME by solving Euler equations, it became possible to determine the sucking force of the ventricle without using any intrusive methods, as stated by Greenberg et al. . Since tTakahashi and colleagues  modified3), v denotes velocity, s denotes position along the color M-mode streamline, and t denotes time. MATLAB is used to obtain v, s, and t and then measure the relative pressures in the region of interest [MATLAB  is used to calculate the IVPD following CMME image acquisition . The folinterest ,21,27. Tinterest .IVPG is calculated by dividing the IVPD by the LV length to exclude the effect of the left ventricular size using the following equation [equation :(2)IVPG A.Rat models of cardiovascular disordersUpdated studies that used CMME-calculated IVPD and IVPG measurements in heart failure are illustrated in murine studies .(a)Changes in IVPD and IVPG in loading conditionsThe current conventional echocardiographic evaluation methods for the assessment of diastolic function have certain recognized limitations, especially under various loading states. The accuracy of conventional echocardiographic methods for diagnosis of DD, such as mitral inflow E-wave velocity (E) using pulsed-wave Doppler echocardiography and mitral annular early diastolic velocity (Em) using tissue Doppler imaging (TDI), is controversial, since they can be influenced by loading conditions with limited efficiency in expressing regional myocardial dysfunction .The effect of loading changes on IVPD and IVPG variables measured through CMME was studied in rats after hydroxyethyl starch and milrinone infusion. The result revealed that mid-to-apical IVPG, mid-IVPG, and apical IVPD were elevated significantly with milrinone infusion; meanwhile, total IVPD and basal IVPD were significantly increased following the infusion of hydroxyethyl starch . The abo(b)Hypertrophic cardiomyopathy (HCM)Advanced heart failure secondary to HCM frequently occurs in humans and cats. Nonetheless, there is still a limited clinical ability of current diagnostic approaches to detect HCM in patients at risk of heart failure ,31. DD iDuring the development of LV hypertrophy in the rat model, total and basal IVPG was increased approximately one month after aortic coarctation , Table 1In veterinary practice, feline HCM is the most frequently detected heart disease in cats; it occurs due to breed predisposition  or due to several genetic mutations. However, for most cats with HCM, the cause is unknown ,34. Enla(c)Diabetic cardiomyopathyThe number of human patients with diabetes mellitus (DM) is rapidly growing in the industrialized world. In DM patients, subclinical LV cardiomyopathy and LA dysfunction usually occur . The patCharacterization of cardiac dysfunction in DM patients usually appears in the advanced stages of the disease. A recent trial found a reduction in longitudinal deformation in all layers and epicardial and mid-myocardial circumferential deformation at the basal level in type-1 DM patients using speckle-tracking echocardiography (STE) . AnotherIn an experimental trial, Kitpipatkun et al. studied the pathophysiologic changes in the IVF in diabetic cardiomyopathic rats, which was experimentally induced via streptozotocin. In this study, diastolic failure and delayed cardiac relaxation were observed in diabetic rats, as indicated by reduced mitral E velocity, E/A ratio, TDI measurements, and mid-to-apical IVPD .(d)Myocardial infarction modelMyocardial infarction (MI) is primarily a type of ischemic heart disease that is caused by an extended shortage of oxygen to the cardiac tissues, which occurs due to constriction or blockage of one or more branches of the coronary circulation ,46. The Two studies have been conducted to evaluate the usefulness of IVPG derived from CMME in MI models in rats , Table 1The second study was conducted to evaluate the efficiency of IVPG to detect subclinical heart failure in rats which were maintained for one month after the induction of MI, as well as to evaluate the efficiency of the new cardioprotective medication, trehalose, on cardiac function . In the (e)Uremic cardiomyopathyIn individuals with renal failure, uremic cardiomyopathy (UC) is traditionally characterized by diastolic dysfunction in conjunction with left ventricular hypertrophy and myocardial fibrosis. ,52. LV hB.IVPD and IVPG in experimental studies in dogs(a)Changes in IVPD and IVPG in loading conditionsMatsuura et al. experimentally sought the IVPD changes under different loading conditions in dogs using transthoracic CMME echocardiography and pressure–volume conductance catheter , Table 2(b)Chemotherapy-induced heart failureBecause DD can occur before the systolic failure, it may be possible to identify diastolic ventricular dysfunction preclinically. In adult cardiac disorders, IVPG is considered an important index of myocardial diastolic function ,9,57. CaThe utility of IVPG was examined for the early identification of subclinical LV dysfunction caused by doxorubicin in Beagle dogs , Table 2C.IVPD and IVPG measurements in the clinical setting in companion animal medicineIVPD and IVPG indices were evaluated in a serial study in dogs with pharmacologically modulated cardiac function as well as normal healthy dogs. Updated studies that used CMME-calculated IVPD and IVPG measurements in companion animals are illustrated in In addition, CMME-measured variables were evaluated in a clinical study of dogs with PDA . PDA is D.IVPD and IVPG in pigsCardiomyopathies, or conditions that affect the cardiac ability to pump blood, can be inherited or acquired; they can manifest as arrhythmogenic, dilated, hypertrophic, ischemic, or restricted conditions. Complex diagnostic standards must be used for these various types of heart failure, and both the pathophysiology and the approach to treatment vary greatly . DilatedThe right ventricular (RV) IVPD measured by CMME in a pig model with changed preload, afterload, and lusitropic states as well as in human patients with DCM has been studied . In pigsE.CMME-derived IVPD and IVPG in sheep and goatsSmall ruminants, especially goats and sheep, serve as good experimental models for cardiovascular research purposes because of the average body size, easy management, and relative heart size to those of humans and companion animals ,75. In aCMME has been studied in sheep and goats. In the sheep model of the LV aneurysm , simultaRecently, the feasibility of CMME-derived IVPD and IVPG measurements in wakeful and sedated goats as well as after pharmacological modulation of the heart was evaluated. Assessment of IVPD and IVPG from CMME was possible in goats and can be used for further cardiovascular experimentations . In goatThe clinical relevance of IVPD and IVPG measured by CMME as promising methods for the early diagnosis of cardiac dysfunction is still under research consideration in laboratory animals, pets, and humans for future validation in clinical cardiology. Characterization of the intracardiac flow dynamics via IVPD and IVPG assessment in feline and canine pediatric cardiology may be important to deepen our understanding of the pathophysiology, prognosis, and follow-up treatment courses, especially when the continuous assessment of the diastolic function is needed.The physiologic variation of IVPD and IVPG in client-owned healthy cats has been established . Future Concerning congenital heart disease (CHD), the evaluation of myocardial function has been widely researched. The systolic function has received the most attention in CHD to date due to its significance and measurement simplicity. However, the diastolic function as an important component of LV function is often overlooked. The conventional parameters which are used to assess diastolic function often measure the blood flow and are affected by the loading conditions of the heart. The problem in diastolic function assessment occurs when attempts are made to evaluate the myocardial relaxation independently of the preload and/or afterload. The interpretation of the diastolic function in the context of CHD requires some understanding of the effects of the lesions themselves on the established diastolic function parameters . CarefulAnother imaging technique, STE, can accurately and quantitatively evaluate diastolic and systolic myocardial performance . A detaiChemotherapeutic agents are known to adversely affect heart function either in an early or latent manner. Few studies demonstrated the efficiency of IVPD and IVPG in chemotherapy-related heart failure in humans and dogs ,62. SincThe current trend in cardiovascular disease diagnostic techniques is aimed at the prediction and early diagnosis of heart failure before the onset of clinical symptoms. For the efficient use of various animals as a candidate for cardiology research, novel echocardiographic techniques are also pertinent for validation in the used model for the accurate interpretation of pathological or pharmacological conditions ,83,84. CThus, the CMME-derived IVPD and IVPG have important theoretical significance and a wide range of application prospects in cardiac disorders as new imaging indices, particularly in the evaluation of diastolic function. In moving away from catheterization, IVPD and IVPG from CMME could be new echocardiographic indices that reflect the diastolic function changes and, in the future, may replace catheterization procedures to diagnose changes in diastolic function. In veterinary medicine, many research trials should be concerned with upcoming studies.