Integrative Physiology Research Articles

HPB O04 - Human Splenic Ex-Vivo Perfusion Model: A Platform for Translational Basic Science Research

Abstract Background The function of the spleen has always been shrouded in mystery and since the Renaissance leading figures have attempted to understand its function. Vesalius questioned Galen and Marcello Malpighi (1628-1694) gave the first accurate description of the histology and the physiology. The spleen still poses unique challenges as invasive approaches may produce intraperitoneal bleeding, differences between human and animal spleens complicates comparative studies, and post-mortem examinations reveal only late-stage disease manifestations and functional studies are impossible. This study describes a human splenic ex-vivo perfusion model for translational research, using microbiological studies as markers of splenic physiological function. Method Patients with left-sided pancreatic cancers scheduled for distal pancreatic and splenic resection were identified from the Hepato-Pancreatic and Biliary MDT at the University Hospitals of Leicester NHS Trust. Patients were informed, consented, and the surgical approach remained standard, with splenic artery ligation deferred until specimen extraction. The artery was cannulated, and residual blood clots were cleared with Custodiol® HTK solution. The perfusion technique employs an extracorporeal circuit incorporateing anoxygenator and a heated water chamber maintaining a 37°C perfusate. The perfused spleen was infected with Streptococcus Pneumoniae strains, with regular biopsies and perfusate gas analysis performed. Results Since February 2023, nine spleens have been successfully perfused, with an average weight of 169.5 grams, cold ischemic time of 116 minutes, and warm ischemic time of 12.2 minutes. Pressure was maintained at 80 mmHg and hourly perfusate gas analysis demonstrated a mean pH of 7.1 and a gradual increase in lactate. Histopathological examination revealed clear distinctions between the lymphoid tissue and splenic red pulp, with intact central arterioles and healthy cell morphology at time 0 and after 6 hours of perfusion. Additionally, the perfused spleens cleared S. pneumoniae strains to below-detection levels. Conclusion Our work demonstrates that a stable human splenic ex-vivoperfusion model which maintains structural and physiological integrity can be achieved from tissue that would otherwise be discarded. The continued physiological function is confirmed by the clearance of S. pneumoniae. While currently employed for microbiological studies, this model has the potential to facilitate a wide range of different translational science studieswhile reducing the need for animal experimentation. The perfusion times employed for the present study were sufficient to demonstrate bacterial clearance which was the aim but there is capacity in the system to significantly prolong the duration.

Diagnostic Validity of Clinical Observations for Detecting Physiologic Swallowing Impairment.

A clinical swallow evaluation (CSE) is a noninvasive and indirect assessment of the anatomical and physiological integrity of the swallowing mechanism in a natural setting. A CSE goes beyond a screening, a dichotomized indicator of dysphagia risk, by comprehensively examining the patient's swallowing through gaining sensory and motor information of oral and pharyngeal function. Information obtained from CSEs in combination with medical, social, and environmental patient related factors allow clinicians to make critical decisions about patients' health and quality of life. It is essential that we have assessment tools with rigorous methodological quality to optimize the accuracy of our clinical judgement. The purpose of this study is to investigate the diagnostic validity of clinical observations obtained through the Mann Assessment of Swallowing Ability (MASA) to better inform clinicians regarding the confidence that the items are testing what they are intended to test and reflect indication of true physiologic swallowing impairment and airway invasion. Area Under the ROC Curve (AUC) analyses revealed that the MASA's diagnostic validity showed acceptable accuracy levels for detecting oral impairment and aspiration, and poor accuracy for detecting pharyngeal impairment and penetration. In the cross-validation analysis, the AUC accuracy level for aspiration changed from acceptable to poor, but remained the same for oral impairment, pharyngeal impairment and penetration. In our sample, acceptable levels for detecting aspiration but poor levels for detecting pharyngeal impairment indicate that the MASA does an adequate job of identifying risk but not explaining the nature of impairment. These results support the need for videofluoroscopic imaging to identify the nature and severity of swallowing impairment, guide intervention and provide recommendations for safe and efficient oral intake.

Long-term effects of a double hit murine model for schizophrenia on parvalbumin expressing cells and plasticity-related molecules in the thalamic reticular nucleus and the habenula

The exposure to aversive experiences during early-life affects brain maturation and induces changes in behavior. Additionally, when these experiences coincide with subtle neurodevelopmental alterations, they may contribute to the emergence of psychiatric disorders, such as schizophrenia. Studies in patients and animal models have identified changes in parvalbumin (PV) expressing inhibitory neurons, highlighting their significance in the etiology of this disorder. Most studies have been focused on the cortex, but PV+ neurons also provide inhibitory input to diencephalic regions, particularly to the thalamus (through cells in the thalamic reticular nucleus, TRN) and the habenula. Remarkably, alterations in both nuclei have been described in schizophrenia. Some of these changes in PV+ cells may be mediated by perineuronal nets (PNN), specialized regions of the extracellular matrix that often surround them and regulate their synaptic input and activity. Interestingly, the physiological maturation and integration of PV+ neurons, which involves the assembly of PNN, occurs during early postnatal life. Plasticity molecules associated to inhibitory neurons, such as PSA-NCAM, or NMDA receptors (NMDAR) can also influence the structure and function of these cells. Growing evidence also indicates that glial cells regulate the physiology of PV+ neurons by influencing their maturation and modulating their synaptic connectivity. To explore the impact of early-life aversive experiences and concomitant subtle neurodevelopmental alterations on diencephalic PV+ cells, we analyzed adult male mice subjected to a double-hit model (DHM) of schizophrenia, combining a single injection of an NMDAR antagonist at P7 and post-weaning social isolation. We observed that exploratory behavior, PV+ neurons and their associated PNN, as well as PSA-NCAM and NMDAR expression and glial cells, in the TRN and the habenula were affected by the DHM or one of its factors. To our knowledge, this is the first report on such alterations in these diencephalic structures in an animal model combining neurodevelopmental alterations and early-life stress during adolescence. Our findings complement previous work on PV+ neurons in cortical regions and underscore the importance of studying diencephalic inhibitory networks and their intricate interactions with aversive experiences and neurodevelopmental alterations during early life in the context of schizophrenia.

Integration of transcriptomics, gut microbiota, and physiology reveals the toxic response of bensulfuron-methyl in Procambarus clarkii

Bensulfuron-methyl (BSM) enters the environment through agricultural practices, posing a threat to the health of aquatic organisms. Currently, the toxic mechanisms of BSM on crayfish (Procambarus clarkii) have not been thoroughly investigated. In this study, crayfish were exposed to BSM solutions at concentrations of 0, 5, and 10 mg/L for 48 h. The study integrated physiological, gut microbiota, and transcriptomic analyses to investigate the mechanisms of action. BSM exposure induced oxidative stress responses in crayfish, resulting in changes in superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GSH) activity, and malondialdehyde (MDA) levels. Exposure to BSM caused damage to the intestinal tissues, reduced gut microbiota diversity, increased the abundance of harmful bacteria, and led to intestinal dysfunction. Analysis of the hepatopancreas revealed significant tissue damage. Transcriptomic data indicated that BSM affects the growth of crayfish through genes related to immune response (SLC17A5, CTSD, CTSB, NFKBIA, Mincle). The lysosomal pathway and NF-κB pathway were notably affected. This study analyzed the negative impacts of BSM on crayfish from various levels and provided detailed data to enhance our understanding of the toxic mechanisms of BSM in aquatic organisms.

Integration of physiology, genomics and microbiomics analyses reveal the biodegradation mechanism of petroleum hydrocarbons by Medicago sativa L. and growth-promoting bacterium Rhodococcus erythropolis KB1

Despite the effectiveness of microbial-phytoremediation for remediating total petroleum hydrocarbons (TPH)-contaminated soil, the underlying mechanisms remain elusive. This study investigated the whole-genome and biological activity of Rhodococcus erythropolis KB1, revealing its plant growth promotion (PGP), TPH degradation, and stress resistance capabilities. Phytoremediation (using alfalfa) and plant-microbial remediation (using alfalfa and KB1) were employed to degrade TPH. The highest TPH degradation rate, reaching 95%, was observed with plant-microbial remediation. This is attributed to KB1′s ability to promote alfalfa growth, induce the release of signaling molecules to activate plant antioxidant enzymes, actively recruit TPH-degrading bacteria (e.g., Sphingomonas, Pseudomonas, C1-B045), and increase soil nitrogen and phosphorus levels, thereby accelerating TPH degradation by both plants and microorganisms. This study demonstrates that R. erythropolis KB1 holds great potential for enhancing the remediation of TPH-contaminated soil through its multifaceted mechanisms, particularly in plant-microbial remediation strategies, providing valuable theoretical support for the application of this technology.

Research on the association mechanism and evaluation model between EMG data and product aesthetic quality in product aesthetics evaluation

Product aesthetics play a crucial role in user satisfaction and product success. However, the feasibility and accuracy of using electromyographic (EMG) data for product aesthetic quality evaluation are unknown. Here, we analysed the correlations and associations between EMG physiological data and product aesthetic quality of 60 subjects by SPSS software and designed a product aesthetic quality evaluation model based on the multivariate logistic (M) algorithm to answer this question. It was found that the EMG data of the zygomatic and buccal muscles did not have significant correlations with product aesthetic quality, whereas five indicators in the EMG data of the frowning muscles had significant correlations relationships with product aesthetic quality. We also found that the overall accuracy of the product aesthetic quality evaluation model was 72.3%, which was able to better distinguish product aesthetic quality. This study demonstrates that product aesthetic quality can be evaluated using EMG data and that the product aesthetic quality evaluation model can provide an objective and accurate decision-making reference to help engineers and designers in product aesthetic quality evaluation and product design. This study lays the foundation for the future integrated multimodal physiology to conduct aesthetics quality evaluations with a high degree of accuracy.

Rapid physiological integration of fused ctenophores

The mechanisms by which organisms recognize the 'self' from the 'non-self' remain poorly understood. Moreover, the capability of transplanted tissue to functionally integrate is unclear in many organisms. Here, we report that two injured Mnemiopsis leidyi individuals, a species of planktonic animals known as comb jellies or ctenophores, are capable of rapidly fusing into a single entity in which some physiological functions are integrated. Our results highlight two interesting phenomena. First, ctenophores may lack an allorecognition mechanism that prevents fusion events between conspecifics. Second, fused individuals rapidly integrate and share physiological functions and neurobehavioral outputs. Ctenophores are among the earliest-branching animal groups of extant metazoans1 and possess a unique nervous system with enigmatic homology to other phyla2. Our observations warrant further research into understanding the evolution of self-nonself recognition systems and the functional integration of neuronal structures in ctenophores.

Ascorbic acid 2-glucoside improves survival, quality, and fertility of frozen-thawed C57Bl/6J and C57Bl/6N mouse spermatozoa.

Ascorbic acid 2-glucoside (AA2G) is a stabilized form of ascorbic acid and a potent antioxidant. Ascorbic acid is present in the testes and epididymis and helps maintain the physiological integrity of reproductive organs. Its properties have been utilized to protect spermatozoa of different species from oxidative stress. Spermatozoa of C57Bl/6J and C57Bl/6N strains were frozen and analyzed, after thawing, by supplementing the capacitation medium with AA2G, both in the presence and absence of methyl-β-cyclodextrin (MBCD). The effect of treatment was evaluated by SCA System (Microptic) analyzing the velocity, vitality, morphology, and the DNA fragmentation. We also examined sperm capacitation (CTC), acrosome reaction (Coomassie Brillant Blue), and fertility (in vitro fertilization) of treated spermatozoa. AA2G improved sperm quality and fertility particularly in association with MBCD. We observed a significant increase of sperm motility, velocity, and vitality associated with an enhanced capacitation and acrosome reaction. These improvements resulted in a marked increase in in vitro fertilization success. Embryos obtained were cultured and reached normally the blastocyst stage. This study aimed to determine if AA2G could safeguard mouse spermatozoa during cryopreservation. We found a protective effect of AA2G that increased sperm survivability resulting in higher fertilization rate. This newly improved protocol shows potential for reanimating cryopreserved GEMMs stored in mouse biobanks and international repositories, such as the European Mouse Mutant Archive (EMMA). This can serve as a pivotal tool in fulfilling the 3Rs mission (replacement, reduction, and refinement), promoting ethical and humane research practices.

Atrial Natriuretic Peptide and Cardiovascular Diseases in Dogs and Cats

The term biomarker encompasses various biological indicators that objectively reflect a patient's medical status with precision and reproducibility. These indicators range from basic measurements like pulse and blood pressure to more intricate laboratory tests. Cardiac markers are crucial for accurate and prompt diagnosis of heart diseases in animals. Given the challenge of diagnosing cardiac diseases in small animals due to nonspecific clinical signs, cardiac markers provide quantitative indicators of biological processes. These markers include cardiac troponins for myocardial injury, natriuretic peptides for myocardial function, lipoproteins for serum homeostasis, and markers for inflammation of the cardiovascular system. Among natriuretic peptides, atrial natriuretic peptide (ANP) has emerged as a significant tool in diagnosing and monitoring cardiac diseases. ANP, primarily synthesized in cardiac atria, regulates salt and fluid excretion, counteracts vasoconstriction, and inhibits the renin-angiotensin-aldosterone system, contributing to the maintenance of cardiovascular homeostasis. Additionally, it functions as a biomarker for ventricular hypertrophy and congestive heart failure (CHF) in animals. Furthermore, it protects against hypertension and cardiac remodeling by demonstrating antagonism to the same system. This review addresses the definition of biomarkers within the context of molecular biology, elucidates their multifaceted functions in the animal organism in light of integrative physiology, and explores the pathologies correlated with ANP, with an emphasis on its etiopathogenesis and clinical manifestations.

Hemodynamic Mechanisms Initiating Salt-Sensitive Hypertension in Rat Model of Primary Aldosteronism.

Few studies have investigated the hemodynamic mechanism whereby primary hyperaldosteronism causes hypertension. The traditional view holds that hyperaldosteronism initiates hypertension by amplifying salt-dependent increases in cardiac output (CO) by promoting increases in sodium retention and blood volume. Systemic vascular resistance (SVR) is said to increase only as a secondary consequence of the increased CO and blood pressure. Recently, we investigated the primary hemodynamic mechanism whereby hyperaldosteronism promotes salt sensitivity and initiation of salt-dependent hypertension. In unilaterally nephrectomized male Sprague-Dawley rats given infusions of aldosterone or vehicle, we found that aldosterone promoted salt sensitivity and initiation of salt-dependent hypertension by amplifying salt-induced increases in SVR while decreasing CO. In addition, we validated mathematical models of human integrative physiology, derived from Guyton's classic 1972 model - Quantitative Cardiovascular Physiology-2005 and HumMod-3.0.4. Neither model accurately predicted the usual changes in sodium balance, CO, and SVR that normally occur in response to clinically realistic increases in salt intake. These results demonstrate significant limitations with the hypotheses inherent in the Guyton models. Together these findings challenge the traditional view of the hemodynamic mechanisms that cause salt-sensitive hypertension in primary aldosteronism. Key words: Aldosterone, Blood pressure, Salt, Sodium, Rat.

Integrative physiology and transcriptome sequencing reveal differences between G. hirsutum and G. barbadense in response to salt stress and the identification of key salt tolerance genes

BackgroundSoil salinity is one of the major abiotic stresses that threatens crop growth. Cotton has some degree of salt tolerance, known as the “pioneer crop” of saline-alkali land. Cultivation of cotton is of great significance to the utilization of saline-alkali land and the development of cotton industry. Gossypium hirsutum and G. barbadense, as two major cotton species, are widely cultivated worldwide. However, until recently, the regulatory mechanisms and specific differences of their responses to salt stress have rarely been reported.ResultsIn this study, we comprehensively compared the differences in the responses of G. hirsutum acc. TM-1 and G. barbadense cv. Hai7124 to salt stress. The results showed that Hai7124 exhibited better growth than did TM-1 under salt stress, with greater PRO content and antioxidant capability, whereas TM-1 only presented greater K+ content. Transcriptome analysis revealed significant molecular differences between the two cotton species in response to salt stress. The key pathways of TM-1 induced by salt are mainly related to growth and development, such as porphyrin metabolism, DNA replication, ribosome and photosynthesis. Conversely, the key pathways of Hai7124, such as plant hormone signal transduction, MAPK signaling pathway-plant, and phenylpropanoid biosynthesis, are mainly related to plant defense. Further comparative analyses of differentially expressed genes (DEGs) revealed that antioxidant metabolism, abscisic acid (ABA) and jasmonic acid (JA) signalling pathways were more strongly activated in Hai7124, whereas TM-1 was more active in K+ transporter-related genes and ethylene (ETH) signalling pathway. These differences underscore the various molecular strategies adopted by the two cotton species to navigate through salt stress, and Hai7124 responded more strongly to salt stress, which explains the potential reasons for the greater salt tolerance of Hai7124. Finally, we identified 217 potential salt tolerance-related genes, 167 of which overlapped with the confidence intervals of significant SNPs identified in previous genome-wide association studies (GWASs), indicating the high reliability of these genes.ConclusionsThese findings provide new insights into the differences in the regulatory mechanisms of salt tolerance between G. hirsutum and G. barbadense, and identify key candidate genes for salt tolerance molecular breeding in cotton.

Aging and physiological barriers: mechanisms of barrier integrity changes and implications for age-related diseases.

The phenomenon of compartmentalization is one of the key traits of life. Biological membranes and histohematic barriers protect the internal environment of the cell and organism from endogenous and exogenous impacts. It is known that the integrity of these barriers decreases with age due to the loss of homeostasis, including age-related gene expression profile changes and the abnormal folding/assembly, crosslinking, and cleavage of barrier-forming macromolecules in addition to morphological changes in cells and tissues. The critical molecular and cellular mechanisms involved in physiological barrier integrity maintenance and aging-associated changes in their functioning are reviewed on different levels: molecular, organelle, cellular, tissue (histohematic, epithelial, and endothelial barriers), and organ one (skin). Biogerontology, which studies physiological barriers in the aspect of age, is still in its infancy; data are being accumulated, but there is no talk of the synthesis of complex theories yet. This paper mainly presents the mechanisms that will become targets of anti-aging therapy only in the future, possibly: pharmacological, cellular, and gene therapies, including potential geroprotectors, hormetins, senomorphic drugs, and senolytics.

Anatomical and physiological responses of roots and rhizomes in Oryza longistaminata to soil water gradients.

Roots and rhizomes are critical for the adaptation of clonal plants to soil water gradients. Oryza longistaminata, a rhizomatous wild rice, is of particular interest for perennial rice breeding due to its resilience under abiotic stress conditions. While root responses to soil flooding are well-studied, rhizome responses to water gradients remain underexplored. We hypothesize that physiological integration of Oryza longistaminata mitigates heterogeneous water deficit stress through interconnected rhizomes, and both roots and rhizomes respond to contrasting water conditions. We investigated the physiological integration between mother plants and ramets, measuring key photosynthetic parameters (photosynthetic and transpiration rate, and stomatal conductance) using an Infrared Gas Analyzer. Moreover, root and rhizome responses to three water regimes (flooding, well-watered, and water deficit) were examined by measuring radial water loss and apparent permeance to O2, along with histochemical and anatomical characterization. Our experiment highlights the role of physiological integration via interconnected rhizomes in mitigating water deficit stress. Severing rhizome connections from mother plants or ramets exposed to water deficit conditions led to significant decreases in key photosynthetic parameters, underscoring the importance of rhizome connections in bidirectional stress mitigation. Additionally, O. longistaminata rhizomes exhibited constitutive suberized and lignified apoplastic barriers, while such barriers were induced in roots under water stress. Anatomically, both rhizomes and roots respond similarly to water gradients, showing thinner diameters under water deficit conditions and larger diameters under flooding conditions. Our findings indicate that physiological integration through interconnected rhizomes helps alleviate water deficit stress when either the mother plant or the ramet is experiencing water deficit, while the counterpart is in control conditions. Moreover, O. longistaminata can adapt to various soil water regimes by regulating anatomical and physiological traits of roots and rhizomes.

Discovery of natural rosin-based preservative candidates to control Phytophthora capsici for postharvest disease management of solanaceae vegetables

To mitigate the postharvest harm of pathogens on solanaceous vegetables, a series of natural rosin-1,2,4-triazole derivatives (totally 24) have been designed and developed. In vivo fungicidal activity evaluation showed that Co. 5 f could inhibit the growth of Phytophthora capsici (EC50 = 0.22 mg/L). In vivo activity suggested that Co. 5 f presented certain protective effect on bell pepper (64.93 % – 32.68 %) and tomato (91.93 % – 67.85 %) fruit. Additionally, Co. 5 f prevented declines in both weight and ascorbic acid levels and reduced the malondialdehyde content of bell peppers and tomato, suggesting its efficacy in vegetable preservation during storage. The physiological research revealed that Co. 5 f disrupted the mycelial cell membrane, accelerated the accumulation of reactive oxygen species, and decreased the enzyme activity of peroxidase (POD) and catalase (CAT) protein activities. Subsequently, quantum chemical calculations and molecular docking successfully predicted the active site of Co. 5 f. Metabolomics was then employed to elucidate the mechanism of physiological disorders, suggesting that differential metabolites were primarily enriched in the biosynthesis of amino acid and secondary metabolites. Transcriptomics was also utilized to further uncover the mechanism of mycelial metabolic disorders, revealing that differential genes were primarily enriched in the biosynthesis of secondary metabolites and amino acid. Additionally, the integration of physiology, metabolomics, transcriptomics, and computer-aided drug design unveiled the action mechanism of Co. 5 f against P. capsici, indicating that the regulation of POD and CAT biosynthesis involved six core genes, while the regulation of ARG, HIS, and TRP biosynthesis involved four core genes. Finally, the expression of these ten core genes was validated using quantitative real-time polymerase chain reaction (qRT-PCR). In summary, the above research integrated physiology, metabolomics, transcriptomics and computer-aided drug design to reveal the action mechanism of Co. 5 f, providing a certain theoretical support for the postharvest disease management of bell pepper and tomato fruit.

State-of-the-art percutaneous coronary intervention for left main coronary artery disease in Japan.

Percutaneous coronary intervention for left main coronary artery disease (LM-PCI) represents a high-risk yet life-saving procedure that has evolved significantly over the years. This review outlines the current state-of-the-art practices for LM-PCI in Japan in detail, emphasizing the integration of coronary physiology and intracoronary imaging alongside with evidence-based standardized technique using latest drug-eluting stents. These advancements enable precise lesion assessment, stent sizing, and optimal deployment, thereby enhancing procedural safety and efficacy. Despite discrepancies between current guidelines favoring coronary artery bypass grafting and real-world practice trends towards increased LM-PCI adoption, particularly in elderly populations with multiple comorbidities, careful patient selection and procedural planning are critical. Future perspectives include further refining LM-PCI through conducting randomized controlled trials integrating advanced techniques and addressing the issue of ostial left circumflex lesions and nationwide standardization of medical care for LM disease.

Investigating the Impact of Senna alata Extract on Hematology and Histopathology of Juvenile of Clarias gariepinus

This paper sheds light on the mechanism of action, potential effects and implication of Senna alata on fish health and ecosystem. Senna alata is renowned for its medicinal properties and ornamental value but is also recognized for its potential toxicity, particularly to aquatic organisms Understanding the toxicity of Senna alata to Clarias gariepinus fingerlings is essential for sustainable aquaculture practices and environmental conservation efforts. 250 juveniles of C. gariepinus were procured from E. I farm Limited in Ikot Ekpene, Akwa Ibom State. Water quality tests were carried out to ensure that optimum water conditions were maintained and that the results obtained were solely due to the effects of the treatments (toxicants) on the fish samples. Chronic toxicity bioassay was done to determine the toxicity of the extract on the blood parameters and tissues of sensitive organs of the exposed fish. The bioassay was designed to be intermediate termed chronic test with 14 days exposure using sub-lethal concentrations of the treatment (0.0gm/, 0.18mg/L, 0.27mg/L. 0.4mg/L, 0.60mg/L, 0.90mg/L). Toxicology dose response (fish mortality) was subjected to detailed analysis using Statistical Package for Social Sciences (SPSS) 22.0. Data were analyzed using descriptive statistics. Senna alata exerted a toxic effect on the fish in the present study and toxicity increased with increased concentration. These findings underscore the potential adverse effects of prolonged exposure to Senna alata on the physiological and cellular integrity of the fish. Intriguingly, the absence of pathological changes in the control groups suggests a distinct response attributable to the treatment. This highlights the importance of continued monitoring and assessment of the impacts of Senna alata on aquatic organisms and ecosystem, emphasizing the need for further research to elucidate the underlying mechanisms and potential long-term consequences.

Comprehensive elucidation of the differential physiological kale response to cytokinins under in vitro conditions

BackgroundKale, a versatile cruciferous crop, valued for its pro-health benefits, stress resistance, and potential applications in forage and cosmetics, holds promise for further enhancement of its bioactive compounds through in vitro cultivation methods. Micropropagation techniques use cytokinins (CKs) which are characterized by various proliferative efficiency. Despite the extensive knowledge regarding CKs, there remains a gap in understanding their role in the physiological mechanisms. That is why, here we investigated the effects of three CKs – kinetin (Kin), 6-benzylaminopurine (BAP), and 2-isopentenyladenine (2iP) – on kale physiology, antioxidant status, steroidal metabolism, and membrane integrity under in vitro cultivation.ResultsOur study revealed that while BAP and 2iP stimulated shoot proliferation, they concurrently diminished pigment levels and photosynthetic efficiency. Heightened metabolic activity in response to all CKs was reflected by increased respiratory rate. Despite the differential burst of ROS, the antioxidant properties of kale were associated with the upregulation of guaiacol peroxidase and the scavenging properties of ascorbate rather than glutathione. Notably, CKs fostered the synthesis of sterols, particularly sitosterol, pivotal for cell proliferation and structure of membranes which are strongly disrupted under the action of BAP and 2iP possibly via pathway related to phospholipase D and lipoxygenase which were upregulated. Intriguingly, both CKs treatment spurred the accumulation of sitostenone, known for its ROS scavenging and therapeutic potential. The differential effects of CKs on brassicasterol levels and brassinosteroid (BRs) receptor suggest potential interactions between CKs and BRs.ConclusionBased on the presented results we conclude that the effect evoked by BAP and 2iP in vitro can improve the industrial significance of kale because this treatment makes possible to control proliferation and/or biosynthesis routes of valuable beneficial compounds. Our work offers significant insights into the nuanced effects of CKs on kale physiology and metabolism, illuminating potential avenues for their application in plant biotechnology and medicinal research.

Automatic semantic segmentation of EHG recordings by deep learning: An approach to a screening tool for use in clinical practice

Background and ObjectivePreterm delivery is an important factor in the disease burden of the newborn and infants worldwide. Electrohysterography (EHG) has become a promising technique for predicting this condition, thanks to its high degree of sensitivity. Despite the technological progress made in predicting preterm labor, its use in clinical practice is still limited, one of the main barriers being the lack of tools for automatic signal processing without expert supervision, i.e. automatic screening of motion and respiratory artifacts in EHG records. Our main objective was thus to design and validate an automatic system of segmenting and screening the physiological segments of uterine origin in EHG records for robust characterization of uterine myoelectric activity, predicting preterm labor and help to promote the transferability of the EHG technique to clinical practice. MethodsFor this, we combined 300 EHG recordings from the TPEHG DS database and 69 EHG recordings from our own database (Ci2B-La Fe) of women with singleton gestations. This dataset was used to train and evaluate U-Net, U-Net++, and U-Net 3+ for semantic segmentation of the physiological and artifacted segments of EHG signals. The model's predictions were then fine-tuned by post-processing. ResultsU-Net 3+ outperformed the other models, achieving an area under the ROC curve of 91.4 % and an average precision of 96.4 % in detecting physiological activity. Thresholds from 0.6 to 0.8 achieved precision from 93.7 % to 97.4 % and specificity from 81.7 % to 94.5 %, detecting high-quality physiological segments while maintaining a trade-off between recall and specificity. Post-processing improved the model's adaptability by fine-tuning both the physiological and corrupted segments, ensuring accurate artifact detection while maintaining physiological segment integrity in EHG signals. ConclusionsAs automatic segmentation proved to be as effective as double-blind manual segmentation in predicting preterm labor, this automatic segmentation tool fills a crucial gap in the existing preterm delivery prediction system workflow by eliminating the need for double-blind segmentation by experts and facilitates the practical clinical use of EHG. This work potentially contributes to the early detection of authentic preterm labor women and will allow clinicians to design individual patient strategies for maternal health surveillance systems and predict adverse pregnancy outcomes.

Learning in context: Undergraduate students' knowledge and the content retention of anatomy between discipline-specific and integrated course approaches.

Undergraduate introductory human anatomy and human physiology courses are either taught as discipline-specific or integrated anatomy and physiology (A&P) sequences. An institution underwent a curricular revision to change the course approach from discipline-specific Human Anatomy and Human Physiology to an integrated A&P I and II sequence, allowing the unique opportunity to explore the potential role of contextual learning in academic achievement and content retention. Mediation and moderation analysis was used to evaluate lecture examinations, laboratory practical examinations, and anatomical content retention between the different course approaches. Undergraduate students in the integrated A&P I course approach performed significantly better on lecture assessments and had a higher anatomy content retention rate at the end of the year than students enrolled in the standalone Human Anatomy course. The lecture examination averages between Human Physiology and A&P II (the second course in the sequence), as well as the anatomy laboratory practical examinations, were not significantly different between discipline-specific and integrated course approaches. The results suggest contextual learning-providing physiological context to anatomical structures-increases the anatomical content retention and academic achievement overall.

Is it the people or the place? The remarkable 30-year period of integrative physiology research in the Carlson Gymnasium at the University of Colorado Boulder.

Historically, programs of physical education and sport were housed in gymnasium buildings on academic campuses. As physical education evolved to the more scientifically focused successor departments of exercise science and kinesiology, faculty specialization developed in the physiology of exercise. With time, some faculty broadened their research to study the integrative physiology of other biological states and stressors. Through this series of events, a small group of integrative physiologists was formed in the Carlson Gymnasium at the University of Colorado Boulder during the 1990s with the goal of conducting novel biomedical research. The challenges were daunting: no contemporary core laboratory facilities, lack of temperature control, piercing external noise, pests, regular flooding, electrical power outages, and lack of funds for renovation. Despite these obstacles, the group established an innovative program of translational physiological research ranging from high-throughput molecular analyses to cell models to rodent studies to clinical trials in humans. These investigators supported their work with grant awards from the National Institutes of Health (NIH), Department of Defense, National Aeronautics and Space Administration (NASA), American Heart Association, and private research foundations totaling ∼$80 M in direct costs from the late 1980s to 2020. Collectively, the faculty and their laboratory personnel published ∼950 articles in peer-reviewed scientific journals. Over that period, 379 undergraduate students, 340 graduate students, 84 postdoctoral fellows, and dozens of junior research faculty received scientific training in Carlson, supported by >$21 M in extramural funding. What was accomplished by this handful of integrative physiologists speaks to the importance of the qualities of the investigators rather than their research facilities in determining scientific success.