Biology

In view of the outstanding progress of machine learning (ML) and growing cost of health systems, it is a current challenge to incorporate artificial intelligence tools into actual medical practice. Here we explored the feasibility and reliability of using machine learning to perform an important immunological investigation that currently requires experienced biologists : Anti-nuclear cytoplasmic antibodies (ANCAs) are important markers for vasculitis and they may be evidenced by microscopic examination of cells labeled with patients' sera. The use of a reliable ML classifier to discriminate between positive and negative samples would increase the rapidity and decrease the cost of immunofluorescence-based ANCA detection. Here, we tested seven well-documented ML algorithms, ranging from simple models such as k nearest neighbors to more complex convolutional neural networks involving millions of adjustable parameter. We studied the feasibility and reliability of classifying 1114 serum samples that had been collected for about 3 years and assayed with conventional procedure. We compared four strategies consisting of assaying either whole microscope fields or individual cell images, and natural images or histograms. The following conclusions were obtained : (i) Several different strategies allowed us to build models stable enough to discriminate between positive and negative samples collected during about 27 months, with a comparison to human classification yielding a kappa index of about 0.7, that may be considered as fairly good and intermediate between the performance of junior and senior biologists. (ii) Simpler ML models combined with theoretical thinking might provide the most rapid and efficient way of developing a reliable test within the framework of a single institution. (iii) In addition, the interpretability of the simplest model provided some theoretical insight into important classification parameters. (iv) An important point and caveat is that the multiplicity and versatility of currently available tools make it an essential requirement to test repeatedly a given model, that must be chosen as simple as possible, to achieve a reliability compatible with medical use. It is concluded that our study provides a strong incentive to incorporate ML tools in well defined medical tests, which might reduce the risk of human errors and pave the way to fully automatic procedures.

In spite of well-established global immune landscape, SARS-CoV-2 is still able to further spread and continue causing infection waves. The current understanding about the reason behind is limited, and it is still difficult to predict the evolution or spreading tread of SARS-CoV-2. Therefore, it is necessary to investigate whether the establishment of population immunity has changed the virus evolution or spreading pattern. In this investigation, one overall analysis of the SARS-CoV-2 spreading in the past several years have been carried out through one thorough genomic epidemiology study, with Germany being chosen as one representative location in view of the systemic efforts for genomic surveillance. The growth advantage of a few predominant variants in its early spreading period has been evaluated through a logistic regression model. The results have revealed that the major circulating SARS-CoV-2 variants since 2023 are mainly derived from the Omicron BA.2 family. Since middle of 2024, most predominant variants were produced primarily through recombination, indicating that the evolution derived from recombination might be the major driving force for the continuous spread of SARS-CoV-2 despite the existence of population immunity. Furthermore, the lower growth advantage of recently emerged variants might possibly lead to a tread of reduction in the frequency of infection wave. The information revealed from this investigation suggests that although short-term spreading tread can be affected by specific virus feature as well as local immunity landscape, the long-term spreading tread is mainly decided by the genomic diversity of the viruses, and can be predicted through phylogenetic and genomic epidemiology investigation. The results have emphasized the importance of maintaining the efforts for genomic surveillance of SARS-CoV-2, which is essential from both medical and research perspectives.

Intraspecific morphological variability presents a complex challenge for biological systematics and biomonitoring, particularly for organisms with high phenotypic plasticity, such as zooplankton. Morphological differences between individuals of the water flea species Bosmina longirostris (Crustacea: Cladocera) are difficult to distinguish visually, parthenogenetic females look morphologically uniform within the species; nevertheless, they demonstrate differences attributable to their geographic origin and developmental stage. A reference dataset of microscopic images was created for the study, including populations from two geographically separated regions (seven ones from European Russia and seven ones from Sakhalin Island in the Pacific Ocean (Far East of Russia) and two age groups, demonstrating the ability of a neural network classify to successfully the intraspecific morphological variation. This study demonstrates that deep learning methods are prospective for the detection and understanding of fine morphological intraspecific differences in the cladocerans.

Toxoplasma gondii is a widespread human pathogen that has multiple, clinically relevant stages in its complex life cycle, including fast-replicating tachyzoites and latent bradyzoites. Bradyzoite differentiation is triggered by stress responses that lead to changes in transcription, translation, and metabolism. Two aspects of this process are addressed in this report: first, whether proteins that play roles in bradyzoite differentiation are specific to T. gondii and other bradyzoite-forming parasites of the Sarcocystidae family, and second, whether new bradyzoite differentiation proteins can be identified in T. gondii. To answer these questions, a phylogenetic approach was used, comparing proteomes of select members of the Sarcocystidae family that form morphologically different bradyzoite cysts and members of the Eimeriidae family that do not form cysts. This approach resulted in 8 distinct clusters of T. gondii proteins that reflected different conservation patterns; for example, one cluster showed conservation among all organisms, while another showed conservation in bradyzoite cyst-forming organisms. Known T. gondii proteins involved in bradyzoite differentiation were found in all clusters, indicating that this process uses both highly conserved pathways as well as bradyzoite-specific pathways. Importantly, the cluster containing proteins that are conserved in bradyzoite-forming organisms contained several known regulators of bradyzoites, and will be a source for identifying novel T. gondii proteins that are involved in bradyzoite differentiation.

Background: Increasing evidence suggests that blood pressure variability (BPV) may offer prognostic value beyond average blood pressure levels. However, data on the association between BPV of ambulatory blood pressure monitoring and mortality in patients aged 80 and older are limited. This study aimed to investigate the relationship between BPV and all-cause mortality in this population. Methods: A total of 5,838 ABPM records from the Geriatrics Department of Beijing Friendship Hospital, collected between October 12, 2018, and June 9, 2025, were analyzed. Patients were divided into death and non-death groups. Subgroup analyses were performed based on the number of completed ABPM sessions. Cox proportional hazards models assessed the associations between BPV and mortality. Kaplan?Meier analysis and log-rank tests were used to compare survival across groups. Results: A median follow-up of 32.0 months included 727 hypertensive patients aged ?80 years. Multivariable cox regression and kaplan?meier analyses showed that the reverse-dipper blood pressure pattern was significantly associated with increased mortality. While short-term BPV was not linked to mortality, greater long-term variability in nighttime SBP and daytime DBP was significantly associated with higher mortality. Conclusion: Among individuals aged 80 and older, those with a reverse-dipper pattern and higher long-term BPV had a significantly higher mortality risk, despite achieving recommended blood pressure targets. Key words: blood pressure variability, ABPM, reverse-dipper pattern, elderly hypertension, mortality

The high efficiency of genome editing presents a challenge when modifying genes associated with viability, welfare, or fertility issues, as implementation of the technology frequently results in mosaic animals with bi-allelic mutations. Combining deactivated Cas9 (dCas9) with Cas9 has been proposed as a strategy to protect one of the two target alleles from editing. We piloted this strategy with 11 genes that are reported as homozygous lethal or associated with welfare issues. We showed that the viability of founders was significantly increased when using 80:20 or 90:10 dCas9:Cas9 ratios, whereas the 70:30 ratio did not yield an equivalent protective effect. The associated overall production rate of mutated founder per manipulated embryo was significantly higher for the 80:20 ratio. Concomitantly, an increased proportion of dCas9 was associated with a significant increase in retention of unedited target alleles but, importantly, did not hinder germline transmission. In addition, editing genes in a paralog cluster with a combination of dCas9 and Cas9 reduced unwanted off-target editing, illustrating a further potential applicability of this approach. This study defines the optimal ratio between dCas9 and Cas9 for strategies aimed at achieving mono-allelic mutations within mosaic founders and proposes a means to reduce the incidence of off-target effects in experiments with limited gRNA options.

AO_SCPLOWBSTRACTC_SCPLOWImproving feed efficiency in pigs is essential for reducing production costs and environmental impacts. This study examines the influence of circadian feeding rhythms and genetic polymorphisms on feed efficiency variability using two pig lines divergently selected for Residual Feed Intake (RFI) over ten generations. Feeding behavior was monitored using automatic concentrate dispensers, recording 6,494,097 visits from 3,824 pigs to analyze meal frequency, duration, and diurnal patterns. LRFI pigs ate less frequently, with larger meals and longer durations, they exhibited two distinct feeding peaks: one around 8:00 AM and a higher one at 5:00 PM and they consumed more feed during the diurnal period and less at night. HRFI pigs showed a smoother, less rhythmic feeding behavior with increased nocturnal intake. The differences between the two RFI lines became more pronounced as the number of generations of selection increased, suggesting a genetic basis. Feeding behaviors, including intake during the two main diurnal peaks, were found to be heritable (heritability estimates: 0.30-0.40) and genetic correlations were observed between feed intake and RFI, especially for intake between the two peaks. Then, we investigated the evolution of allele frequencies of single nucleotide polymorphisms (SNPs) in DNA sequences surrounding 10 core clock genes (ARNTL, CLOCK, CRY1, CRY2, NPAS2, NR1D1, PER1, PER2, PER3, RORA) along generations of selection. SNPs with significant frequency changes were mapped to regulatory regions and transposable elements, especially in HRFI line, suggesting potential functional impacts on circadian regulation. These results underscore the role of feeding behavior and genetic variation in feed efficiency, offering insights for breeding programs aimed at improving metabolic efficiency and sustainability in pig production.

Boar semen contains spermatozoa and seminal plasma (SP) that carries extracellular vesicles (EVs) among other components. However, artificial insemination (AI) doses produced by AI companies are highly diluted based solely on sperm concentration. The aim of this study was to evaluate the integrity of EVs isolated from AI doses, characterize the protein and miRNA content from high-fertility (HF) and reduced-fertility (RF) boars, and evaluate their functional impact on spermatozoa after dilution by a coincubation up to 24 hours at 38 {degrees}C. Proteomics identified 108 differentially expressed proteins between HF and RF EVs (97 upregulated in HF, 11 in RF), and transcriptomics revealed 80 differentially expressed miRNAs (DEMs) in EVs, 52 in SP, and 3 in spermatozoa, showing inverse expression in various shared DEMs between fertility rates, suggesting compartment-specific regulation. Functional coincubation demonstrated that EVs remain biologically active after dilution. HF EVs improved sperm quality parameters and reduced oxidative stress, while RF EVs increased total and progressive motility. Overall, our findings show that EVs from AI doses retain structural integrity, carry fertility-associated protein and miRNA signatures, and functionally modulate sperm quality in vitro. These features highlight porcine EVs as promising biomarkers and potential tools to optimize reproductive performance in swine production.

Bdelloid rotifers are known to survive desiccation and high doses of ionizing radiation. This extreme resistance is notably due to their capacity to cope with numerous DNA double-strand breaks (DSBs). Genes encoding key components of the non-homologous end joining (NHEJ) DNA repair pathway are strongly upregulated in the bdelloid rotifer Adineta vaga following exposure to ionizing radiation. Considering the notably high doses tolerated by these organisms, their capacity to efficiently restore genome integrity is particularly striking. Although NHEJ is generally regarded as less accurate than homologous recombination (HR), the absence of major genomic rearrangements in the descendants of irradiated rotifers suggests that DNA repair occurs with high fidelity. Terwagne et al. recently reported a delayed repair in germline nuclei, occurring during oocyte development when homologous chromosomes pair, thereby enabling template-based repair through HR. In this study, we established an in situ hybridization approach on A. vaga cryosections to investigate the spatial and temporal expression of key actors involved in NHEJ, HR, and Base excision repair (BER) pathways in somatic and germline tissues. We show that NHEJ (KU80) and BER-related genes (PARPs) as well as A. vaga Ligase E (putatively involved in DNA repair) are expressed early after radiation exposure in the somatic syncytium. In contrast, HR-related genes (Rad51: two paralogs, Rad54), as well as PCNA (involved in DNA replication, NER, BER, HR) are expressed later in maturing oocytes, indicating the activation of a delayed homologous recombination repair pathway in germline nuclei. Nurse cells, which express genes associated with both HR and NHEJ pathways, may rely on both mechanisms for their own DNA repair while also supplying mRNAs to the maturing oocyte. Our results provide new evidence for a differential regulation of DNA DSB repair pathways between soma and germline in bdelloids, with NHEJ predominating in somatic tissues and HR in the germline of A. vaga. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/722046v1_ufig1.gif" ALT="Figure 1"> View larger version (35K): org.highwire.dtl.DTLVardef@3b1f3borg.highwire.dtl.DTLVardef@17f5eb5org.highwire.dtl.DTLVardef@122ef14org.highwire.dtl.DTLVardef@7e4413_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOAbstract Figure:C_FLOATNO Summary of in situ hybridization results: genes coding for actors of NHEJ are expressed in the somatic nuclei and in the nurse nuclei of Adineta vaga individuals 2.5 hours post X-rays radiation, while genes coding for HR actors and PCNA (involved in multiple pathways including DNA replication and DNA repair: NER, BER, MR, HR) are expressed in the nurse nuclei 2.5 hours post radiation, and later in the maturing oocyte during oogenesis and in the laid eggs. Genes coding for actors highly expressed post-radiation, involved in the BER pathway appear to be only expressed in the somatic syncytium 2.5 hours post radiation, as well as the gene coding for the Ligase E, likely involved in DNA repair. C_FIG

Alleles of ASIP gene (Agouti locus) in dogs determine a wide spectrum of coat colors, from red to black. Gain-of-function Ay allele is the most dominant in the range of known ASIP mutations: when all other genes affecting coat pigmentation are intact, presence of Ay allele results in red coat color. Loss-of-function a allele is the most recessive allele of this gene. When homozygous, it gives black coat color. Usually, dogs with Ay/a genotype have red coat, because a single copy of Ay allele is sufficient to fully compensate for the non-functional allele a, implying the complete dominance in this pair of alleles. However exceptions are known. In the Hungarian Puli breed there is a specific coat pigmentation type called fako. We investigated the genetic composition of fako dogs and found evidence that the dominance of the Ay allele over the a allele may be incomplete in these dogs. Analysis of the MC1R gene that interacts with ASIP in the hair pigmentation genetic cascade allowed us to find the variants that may be responsible for the incomplete dominance of Ay allele over a allele in Hungarian Puli dogs.

Documenting and understanding the welfare of aging animals are crucial for maintaining their well-being and making appropriate management decisions. This study details the behaviors of an extremely old rhesus macaque (ISK) in which senile plaques and phosphorylated tau deposition were observed in post-mortem pathological analyses of the brain. We report on the activity bsudgets, behavioral rhythms, gait, quality of life (QoL) scores, and anecdotal episodes of this individual. The average 24-hour activity budgets, analyzed from surveillance camera recordings, revealed that ISK spent most of her time inactive. ISK was sometimes active at night, though her behavior remained predominantly diurnal. Gait analysis suggested that her movement patterns changed between the first (December 2020) and the last (June 2021) assessment. QoL assessments, using a scoring sheet, indicated relatively good well-being until the later stage of her life. An anecdotal episode, along with the husbandry diary, suggested signs of cognitive decline. These results suggest possible signs of physical decline, and some behavioral changes that could be associated with cognitive decline in an extremely old rhesus macaque. However, we could not confirm cognitive dysfunction without further controlled cognitive testing. We hope that future studies will consider the behavioral symptoms observed in this study as monitoring items to better understand physical and cognitive decline, and possible relationships with QoL in primates.

Anthracyclines, including doxorubicin, are widely used chemotherapeutic agents, but dose-dependent cardiotoxicity limits their clinical utility and increases the risk of heart failure in cancer survivors. In paediatric patients, female sex is a significant risk factor for anthracycline-associated cardiotoxicity, yet pre-clinical studies rarely investigate sex differences in immature hearts. Here, we provide a proteomic dataset from primary cardiomyocytes, isolated from postnatal day 2 rat hearts and treated with a clinically relevant concentration of doxorubicin. Analysis of proteins present in all samples identified candidates previously shown to be regulated by doxorubicin in adult hearts, as well as candidates that may be specifically regulated in young hearts. This dataset provides a resource for generating hypotheses on molecular mechanisms contributing to sex differences in juvenile doxorubicin-induced cardiotoxicity.

Cancer-related genomic instability (GI) may cause genetic alterations in spermatozoa, implying health issues not only in cancer survivors, but also in their children [1, 2]. We therefore studied Loss of Y chromosome (LOY), considered as hallmark of GI [3-15], in spermatozoa and blood from survivors of childhood and testicular cancer (CC, TC), and controls (CTRL). We found that LOY was statistically significantly more frequent in spermatozoa from cancer survivors than in controls (Odds Ratio [OR]=2.2 for CC vs. CTRL and OR=2.4 for TC vs. CTRL). Furthermore, LOY was about an order of magnitude more prevalent in spermatozoa than in blood among 18-53-year-old males within all cohorts. Our findings suggest that LOY in spermatozoa might be a clinically useful marker of GI, reduced fertility and disease predisposition in males. Introducing LOY in spermatozoa as a biomarker opens a new research avenue into disease prevention and the causes and consequences of LOY.

This study presents a proof-of-concept cyber-physical architecture integrating a SEIR epidemiological model (Susceptible-Exposed-Infectious-Recovered), implemented in MATLAB, with a simulated Internet of Things (IoT) acquisition and transmission stage based on the ESP32 microcontroller and the ThingSpeak platform. The system generates synthetic biomedical signals of body temperature and peripheral oxygen saturation (SpO2), structured across three levels: circadian variation, scheduled pathological episodes, and Gaussian noise. These signals feed a dual parametric coupling function that dynamically updates the SEIR transmission parameter as a combined function of body temperature and oxygen saturation deviations from their clinical reference values. The proposed architecture is organized into four functional phases: measurement, communication, computational processing, and feedback. Five simulated clinical scenarios were evaluated, ranging from normal conditions (T = 36.5 {degrees}C, SpO2 = 97%) to fever with severe hypoxia (T = 38.5 {degrees}C, SpO2 = 88%), yielding basic reproduction number (R0) values between 4.20 and 5.38, and peak infected proportions between 29.9% and 35.2% of the simulated population (N = 1,000). A sensitivity analysis on the coupling coefficients, with {+/-}50% variation from nominal values, showed that the oxygen saturation coefficient is the most influential parameter on R0 (range = 0.76) compared to the thermal coefficient (range = 0.42), with monotonic and predictable behavior across the entire evaluated parametric space. The primary contribution of this work is system integration: we propose a reproducible platform connecting biomedical simulation, IoT communication, and epidemiological modeling through parametric coupling in a controlled environment. All data used are entirely synthetic; a retrospective calibration with real Colombian data from the first epidemic wave of 2020 confirmed the epidemiological consistency of the model, with a calibrated R0 of 1.85 and a Pearson correlation of 0.930. Results should be interpreted as evidence of architectural feasibility, not as clinical or epidemiological validation. Author SummaryThe COVID-19 pandemic made it clear that epidemiological surveillance systems need tools that combine accessible technology with mathematical models capable of anticipating disease spread. In this work, we built a proof-of-concept platform connecting three elements: a low-cost electronic sensor based on the ESP32 microcontroller, a cloud communication platform (ThingSpeak), and a mathematical model that simulates how an epidemic spreads through a population. The sensor generates synthetic data on body temperature and oxygen saturation that, through a mathematical formula we designed, dynamically modify the rate of contagion in the model. We evaluated five clinical scenarios, ranging from normal conditions to fever with severe hypoxia, and analyzed how sensitive the results are to changes in the system parameters. We found that oxygen saturation has a greater influence on the estimated contagion potential than body temperature. Although all data are synthetic, this platform demonstrates that it is possible to integrate low-cost sensors with epidemiological models in real time, opening a viable pathway for early warning systems in resource-limited settings.

Vestibular neurons are core elements of the pathways involved in vestibulo-motor functions, such as vestibulo-spinal and vestibulo-ocular reflexes. To meet behavioral needs, electrophysiological neuronal properties are adequately adapted to the sensory-motor computation sustaining these distinct vestibular reflexes. During frog metamorphosis, there is a complete reorganization of the posturo-locomotor system while the oculomotor system remains minimally changed, probably associated to so far unknown changes in vestibular neuronal properties. We used this unique model to investigate the central developmental mechanisms underlying such a reconfiguration of vestibular-associated behaviors. Central vestibular neurons exhibit two types of electrophysiological phenotypes: tonic neurons with a continuous discharge and phasic neurons with a transitory discharge mainly due to the activation of Kv1.1 channel. Electrophysiological recordings and Kv1.1 immunolabeling of vestibulospinal (VS) and vestibulo-ocular (VO) neurons at both larval and juvenile stages revealed that the majority of VS neurons exhibited a tonic discharge in larvae but a phasic discharge in juvenile, while VO neurons remained mainly tonic throughout development. Changes in phasic and tonic neurons proportions in VS population are partly explained by neurogenesis. But we provide evidences that an electrophysiological phenotype switch is a concomitant developmental mechanism participating in the maturation of these central vestibular neurons. All together our results showed that the maturation process in central vestibular neuronal groups is highly related to the metamorphosis-induced remodeling of vestibulo-motor functions they are involved in, with the ultimate purpose of ensuring an adequate adaptation of neuronal elements properties to the developmental changes of behavioral constrains.

Whole genome duplication is a common mutation in eukaryotes with far-reaching phenotypic effects. The resulting morphological, physiological, and fitness consequences and how they affect the survival probability of newly polyploid lineages are intensively studied, but very little is known about the effect of genome doubling on the short-term evolvability of populations. Understanding the effect of polyploidization on the adaptive potential of populations is of crucial importance to predict the future of polyploid populations. In this paper, I investigate the immediate consequences of genome doubling on the genetic variance of populations. To do so, I performed numerical iterations and simulations of how the genetic variance of a quantitative trait changes after polyploidization, under different genetic architectures (additivity, dominance, and epistasis). I found that genetic variance generally decreases after genome doubling. Non-additive gene actions can make autotetraploid populations genetically more diverse than their diploid progenitors in rare cases, notably with overdominance and directional epistasis. By collecting estimates from the agronomic literature, I found that both dominance and epistatic variance contribute to the genetic variance of polyploid populations. These results bring new insights into the adaptive potential of newly formed tetraploid populations, and call for further experimental investigations of how polyploidization is associated with a short-term decrease in evolvability.

This work focuses on the global and partial identification problem for fractional differential equations. We provide a general numerical procedure based on global and local optimization algorithms with two refinements for biological systems that ensure solution positivity and homogeneous parameter units. The method is applied to a new fractional model of Dengue outbreak called the Fractional Homogeneous Nishiura (FHN) model, calibrated using data of newly infected people in Cape Verde. We show that our identification method yields a better fit between data and model solutions than previous approaches and that our FHN model captures the dynamics of Dengue more closely than existing systems.

The Drosophila adult central brain contains 240 circadian neurons, of which there are more than 25 different neuron subtypes based on connectomic data. Recent single cell RNA-seq (scRNAseq) characterization of these neurons "around the clock" also indicates a similar number of molecular subtypes of circadian neurons, but other conclusions from these transcriptomic studies warranted verifying and extending with other approaches. To this end: 1) We used a genetic multiplexing strategy to profile the transcriptomes of circadian neurons from multiple time points in a single experiment, reducing confounding technical variation between timepoints; 2) Large numbers of single nuclei were sequenced (snRNA-seq), which was enabled because the new method EL-INTACT purifies nuclei from frozen heads; 3) We assayed 12 time points under both light-dark (LD) and constant darkness (DD) conditions. These approaches showed dramatic transcriptional differences between time points in many circadian neuron types and enhanced time-of-day gene expression analysis. The data indicate that most of this regulation is transcriptional and circadian. There were however a small number of light-dependent transcripts, including a few that correspond to mammalian immediate-early genes. They probably play a role in the light-regulation of gene expression and behavior in specific neurons, perhaps circadian entrainment or phase-shifting. The results taken together provide a more comprehensive picture of gene expression heterogeneity within adult Drosophila circadian neurons including how intrinsic clock mechanisms and light cues are integrated across circadian neuron subtypes.

The paper presents the results of a comparative analysis of gene networks activated by water stress and low temperatures in extensive (Saratovskaya 29, S29) and intensive (Yanetskis Probat, YP) wheat varieties during the seedling development stage. It is concluded that the creation of the S29 variety, which occurred through complex stepwise hybridization and selection for morphological traits, productivity, and grain quality traits, resulted in the emergence and inheritance of regulatory gene networks involving proteins with the CC domain, as well as the BTB/POZ and TAZ domains, which have an increased affinity for transcription factors involved in the transcriptomic response to changing external conditions. It was established that, at the transcriptomic level, the S29 variety is characterized by a transition to an energy saving mode to maintain the activity of the Calvin-Benson cycle under the water deficit conditions and the inhibition of proteolytic processes at low temperatures. The transcriptional response of the high-yielding YP variety to 24-hour low-temperature treatment was more active and involved a larger number of genes compared to the S29 variety. Identifying varietal variability in molecular genetic mechanisms of resistance to abiotic stressors facilitates the development of marker-assisted and genomic selection technologies for common wheat. Key messageThe extensive S29 variety was characterized by its transition to energy-saving mode to maintain the Calvin-Benson cycle under water deficit and a reduction in proteolytic processes under low temperature.

Gestational diabetes mellitus (GDM) is a major metabolic complication of pregnancy with significant maternal and fetal adverse consequences. Beyond classical mechanisms, emerging evidence suggests that gut-derived metabolic endotoxemia may contribute to dysglycemia. Intestinal alkaline phosphatase (IAP), a key enzyme involved in maintaining gut barrier integrity and detoxifying lipopolysaccharides, has been linked to type 2 diabetes mellitus; however, its role in GDM remains largely unexplored. This hospital-based cross-sectional analytical study evaluated fecal IAP levels and their association with GDM among 198 pregnant women recruited from three antenatal care clinics representing three tiers of ANC services. Participants were screened for GDM using a 75-g oral glucose tolerance test and classified as having GDM (n=55) or normal glucose tolerance (NGT; n=143) according to WHO 2013 criteria. Stool samples were collected, and fecal IAP levels were measured using an enzymatic colorimetric assay. Fecal IAP level was significantly lower in women with GDM than in those with NGT (median 23.59 vs 46.48 U/g stool; p<0.001). Lower IAP level remained independently associated with GDM after adjustment for body mass index and previous GDM (adjusted OR 0.98 per unit increase; 95% CI 0.97-0.99; p<0.001). A graded relationship was observed between declining IAP level and GDM. Receiver operating characteristic analysis demonstrated modest discrimination (AUC 0.676), while a threshold of approximately 65 U/g stool yielded high sensitivity (89.1%) but lower specificity. Reduced fecal IAP is independently associated with GDM, supporting a potential role of gut-derived metabolic dysregulation in gestational glucose intolerance. While not suitable as a standalone diagnostic tool, fecal IAP may serve as a complementary biomarker for risk stratification during pregnancy. Prospective studies are warranted to determine its predictive value and explore its potential as a therapeutic target.