Abnormal Development Research Articles

Development of a TBXT-EGFP iPS Cell Model for Screening the Early Developmental Toxicity of Typical Environmental Pollutants

In our daily lives, we are inevitably exposed to a variety of environmental pollutants in numerous ways. Fortunately, recent years have witnessed significant advancements in the field of stem cell toxicology, which have provided new opportunities for research in environmental toxicology. Applying stem cell technology to environmental toxicology, overcomes some of the limitations of traditional screening methods and we can more accurately predict the toxicity of environmental pollutants. However, there are still several aspects of stem cell toxicology models that require improvement, such as increasing the throughput of detection and simplifying detection methods. Consequently, we developed an environmental pollutant toxicity detection model based on TBXT-EGFP iPS cells and screened the developmental toxicity of 38 typical environmental pollutants. Our results indicate that TBBPA-BDBPE, TBBPA-BHEE, DG, and AO2246 may interfere with the expression of TBXT, a critical marker gene for early human embryo development, implying that these environmental pollutants could lead to developmental abnormalities.

Reduced Sox7 contributes to abnormal valve development resulting in congenital aortic stenosis

Abstract Background Abnormal valve development is the most common congenital heart malformation. During valve remodeling, abnormal changes in valve cell proliferation, apoptosis, and extracellular matrix synthesis affect valve development, resulting in malformations. The molecular mechanisms underlying pathophysiology of aortic stenos is remain unclear. Purpose We would like to explore how Sox7 is involved in aortic valve development. Methods and Results We proved Sox7 was mainly expressed in endocardial and mesenchymal cells of aortic valves by immunofluorescence staining determined by at different stages of mice (E10.5～8 weeks) and human embryos and RNA-seq of E9.5 mouse embryonic hearts. We constructed a conditional Sox7 floxed allele in endothelial cells using an endocardial specific Cre. Echocardiography data showed that Nfat-KO mice had increased blood velocity, pressure gradient across aortic valves, left ventricular mass and dilated aortic roots. Then, HE and Sirius red staining to examine histological changes in aortic valves of Nfat-KO mice were performed, and we observed a remarkable increase in the size and increased staining of fibrotic cells in aortic valves of KO mice. Next, we collected embryonic hearts of Sox7 CNC-specific knockout mice (WNT1-KO) and controls at E16.5. According to HE staining results and echocardiography analysis, there was no difference in aortic valves between WNT1-KO mice and controls. Meanwhile, we collected Nfat-KO mice and control embryos at E11.5 days. HE staining results showed that EndMT process was not impaired. Interestingly, leaflets and cusps of Nfat-KO mice showed significantly augmented proliferation levels, decreased apoptosis levels, and calcium deposition in fibrin layer. A significant decrease in MMP9 levels were observed in both mRNA and protein levels. Co-transfection of Sox7-expressing vector and MMP9 luciferase in MEEC suggested that Sox7 significantly activated MMP9. We used adenovirus to overexpress Sox7 in valvular interstitial cells. Compared with that in control group, the Sox7-overexpression group increased protein level of MMP9 and cleaved caspase-3, and increased Tunel-positive rate and lower Ki-67 positive rate. Finally, we detected expression of Sox7 protein in human fetal aortic valves derived from patients with aortic stenosis using immunofluorescence. The results demonstrated that the number of Sox7 positive mesenchymal cells in human fetal aortic valves of patients was significantly lower than that in control group. Conclusion We demonstrated that targeted inactivation of Sox7 in the endocardium of mice affects valve development and leads to aortic stenosis. Mechanistically, no influence of Sox7 on the EndMT process of the OFT cushion. Moreover, CNC-derived Sox7 was not involved in aortic valve development. Interestingly, we found that Sox7 regulated ECM remodeling through matrix MMP9 and Sox7 had an impact on apoptosis, proliferation, and calcification.

Anemia in patients with chronic kidney disease and its association with cardiac abnormalities

Abstract Introduction Chronic kidney disease (CKD) is a pathology characterized by multisystemic comorbidities, predominantly associated with cardiovascular disease and anemia. However, the precise association between the degree of anemia and the development of cardiac abnormalities in patients with CKD undergoing hemodialysis remains unclear. Purpose To determine the association between the degree of anemia and cardiac abnormalities in patients with CKD receiving hemodialysis treatment. Methods A cross-sectional multicenter study that included individuals with CKD undergoing hemodialysis was conducted. Electronic medical records containing information regarding medical conditions and pertinent laboratory parameters was collected, and cardiac function was determined using echocardiography. A Spearman rank correlation analysis was conducted to evaluate the associated between the degree of anemia and echocardiographic parameters, with significant set at p <0.05. Results Ninety-nine individuals were included, with a median age of 48 years. The population was categorized based on hemoglobin (Hb) levels into non/mild anemia (Hb >10 mg/dL) and moderate/severe anemia (Hb <9.9 mg/dL) groups (table). Significant differences were observed in left ventricle (LV) posterior wall thickness (10mm in non/mild vs. 11mm in moderate/severe, p<0.015), left atrial (LA) volume (34 mL/m2 in non/mild vs. 39 mL/m2 in moderate/severe, p<0.007) and left ventricular ejection fraction (LVEF) (59% in non/mild vs. 55% in moderate/severe, p<0.025). Negative correlations (figure) were observed between Hb levels and LV mass index (r= -0.21, p= 0.03) and pulmonary artery systolic pressure (PASP)(r= -0.27, p= 0.006). Conclusion The degree of anemia demonstrated an association with cardiac abnormalities, as evidenced by alterations in echocardiographic parameters such as LVEF, LA volume and PSAP. This study underscores the importance of further prospective investigations to enhance early identification and stratification of anemia in patients with CKD aiming to mitigate further cardiac complications.Clinical demographics tableScatter plots with correlation graphs

Nicorandil activates lkb1-ampk signaling pathway to ameliorate cardiac remodeling after myocardial ischemia/reperfusion injury by upregulating mt2

Abstract Background Cardiomyocyte apoptosis is a crucial event underlying the development of cardiac abnormalities and dysfunction after myocardial ischemia reperfusion (MI/R) injury. A better understanding of the cell signaling pathways involved in cardiac remodeling may support the development of new therapeutic strategies for the treatment of heart failure (HF) after MI/R. Nicorandil, an ATP-sensitive potassium channel opener, could improve mitochondrial damage and reduce oxidative stress，which reduce cardiomyocyte apoptosis. Metallothionein (Mt), reactive oxygen species (ROS) scavenger, which can reduce oxidative stress, also attenuated MI/R-induced autophagy and cell apoptosis. Except as a potassium channel opener, nicorandil may play a cardioprotective role by regulating metallothionein, and its specific mechanism remains to be elucidated. Objective To clarify the protective effect of nicorandil in myocardial ischemia/reperfusion injury, and elucidate the specific mechanism which nicorandil regulate the expression of metallothiosin to reduce myocardial ischemia/reperfusion injury, so as to provide new drug targets or new measures for clinical prevention and treatment of MI/R. Methods A cardiac MI/R injury model was constructed by the ligation of the left anterior descending coronary artery to investigate the underlying molecular mechanisms. The apoptosis of myocardial tissue cells was detected by TUNEL staining. The ultrastructural changes of myocardial tissue were observed by transmission electron microscopy. The expressions of Mt family, mitochondrial dynamics and Glucose metabolism-associated genes were measured by Western blotting and qPCR. The content of ATP was determined by ELISA in heart tissue. AutodockVina was used to evaluate binding energy and interaction patterns between drug candidates and their targets. The protein-protein interaction model was predicted by ZDOCK software. Results Treatment with nicorandil mitigated left ventricular enlargement, improved cardiac dysfunction,reduced myocardial infarcation area and decreased cardiomyocyte apoptosis after I/R. Nicorandil up-regulated the expression of Mt2 and decreased the expressions of Drp-1,p-Drp-1(ser616), while enhanced the expression of Mfn1/2,GLUT4,HK II and MPC II in myocardium. Knockdown of Mt2 decreased the effects of nicorandil on apoptosis and mitochondrial dysfunction after I/R. Mechanistically, nicorandil significantly upregulated the expression of Mt2, which could bound to LKB1 and cause phosphorylation of LKB1, resulting in AMPK-dependent activation. Conclusions Nicorandil significantly upregulated the expression of Mt2, which interacted with LKB1 to promote LKB1 protein phosphorylation, resulting in AMPK-dependent activation and subsequently alleviating I/R-induced mitochondrial dysfunction and energy metabolism disorder, and thereby reducing cardiomyocyte apoptosis, eventually improving I/ R-induced cardiac remodeling and dysfunction.

Case Report: Management of a congenital intrahepatic portosystemic shunt with portal vein aneurysm in a child using 3D computer-assisted partial right hepatectomy

Congenital portosystemic shunts (CPSS) are rare pediatric vascular malformations characterized by abnormal development of the portal vein, which is attributed to incomplete embryonic remodeling of the hepatic and surrounding vasculature. CPSS manifests in two main forms: intrahepatic and extrahepatic. This study details the management of a pediatric patient diagnosed with Congenital Intrahepatic Portosystemic Shunt (CIPS) who was referred to our institution. By using a computer-assisted surgical system, the right hepatectomy was successfully performed, guided by precise intraoperative navigation based on three-dimensional reconstructions of enhanced CT imagery. The patient exhibited a favorable postoperative recovery trajectory, with the absence of complications or recurrence throughout the monitoring period.

Mouse models of Slc35a2 brain mosaicism reveal mechanisms of mild malformations of cortical development with oligodendroglial hyperplasia in epilepsy.

Brain somatic variants in SLC35A2 were recently identified as a genetic marker for mild malformations of cortical development with oligodendroglial hyperplasia in epilepsy (MOGHE). The role of SLC35A2 in cortical development and the contributions of abnormal neurons and oligodendrocytes to seizure activity in MOGHE remain largely unexplored. Here, we generated a novel Slc35a2 floxed allele, which we used to develop two Slc35a2 conditional knockout mouse lines targeting (1) the Emx1 dorsal telencephalic lineage (excitatory neurons and glia) and (2) the Olig2 lineage (oligodendrocytes). We examined brain structure, behavior, and seizure activity. Knockout of Slc35a2 from the Emx1 lineage, which targets both cortical neurons and oligodendrocytes, resulted in early lethality and caused abnormal cortical development, increased oligodendroglial cell density, early onset seizures, and developmental delays akin to what is observed in patients with MOGHE. By tracing neuronal development with 5-Ethynyl-2'-deoxyuridine (EdU) birthdating experiments, we found that Slc35a2 deficiency disrupts corticogenesis by delaying radial migration of neurons from the subventricular zone. To discern the contributions of oligodendrocytes to these phenotypes, we knocked out Slc35a2 from the Olig2 lineage. This recapitulated the increased oligodendroglial cell density and resulted in abnormal electroencephalographic activity, but without a clear seizure phenotype, suggesting Slc35a2 deficiency in neurons is required for epileptogenesis. This study presents two novel Slc35a2 conditional knockout mouse models and characterizes the effects on brain development, behavior, and epileptogenesis. Together, these results demonstrate a direct causal role for SLC35A2 in MOGHE-like phenotypes, including a critical role in neuronal migration during brain development, and identify neurons as key contributors to SLC35A2-related epileptogenesis.

Photosynthetic characteristics and genetic mapping of a yellow-green leaf mutant jym165 in soybean

BackgroundLeaves are important sites for photosynthesis and can convert inorganic substances into organic matter. Photosynthetic performance is an important factor affecting crop yield. Leaf colour is closely related to photosynthesis, and leaf colour mutants are considered an ideal material for studying photosynthesis.ResultsWe obtained a yellow-green leaf mutant jym165, using ethyl methane sulfonate (EMS) mutagenesis. Physiological and biochemical analyses indicated that the contents of chlorophyll a, chlorophyll b, carotenoids, and total chlorophyll in the jym165 mutant decreased significantly compared with those in Jiyu47 (JY47). The abnormal chloroplast development of jym165 led to a decrease in net photosynthetic rate and starch content compared with that of JY47. However, quality traits analysis showed that the sum of oil and protein contents in jym165 was higher than that in JY47. In addition, the regional yield (seed spacing: 5 cm) of jym165 increased by 2.42% compared with that of JY47 under high planting density. Comparative transcriptome analysis showed that the yellow-green leaf phenotype was closely related to photosynthesis and starch and sugar metabolism pathways. Genetic analysis suggests that the yellow-green leaf phenotype is controlled by a single recessive nuclear gene. Using Mutmap sequencing, the candidate regions related of leaf colour was narrowed to 3.44 Mb on Chr 10.ConclusionsAbnormal chloroplast development in yellow-green mutants leads to a decrease in the photosynthetic pigment content and net photosynthetic rate, which affects the soybean photosynthesis pathway and starch and sugar metabolism pathways. Moreover, it has the potentiality to increase soybean yield under dense planting conditions. This study provides a useful reference for studying the molecular mechanisms underlying photosynthesis in soybean.

Familial osteochondrodysplastic and cardiomyopathic syndrome in Chianina cattle.

Skeletal dysplasia encompasses a heterogeneous group of genetic disorders characterized by an abnormal development of bones, joints, and cartilage. Two Chianina half-sibling calves from consanguineous mating with congenital skeletal malformations and cardiac abnormalities were identified. To characterize the disease phenotype, to evaluate its genetic cause, and to determine the prevalence of the deleterious alleles in the Chianina population. Two affected calves, their parents and 332 Chianina bulls. The affected animals underwent clinicopathological investigation. Whole-genome sequencing trio-approach and PCR-based assessment of the frequency of TDP-glucose 4,6-dehydratase (TGDS) and laminin subunit alpha 4 (LAMA4) alleles were performed. The cases presented with retarded growth, poor nutritional status associated with muscular atrophy and angular deformities of the hindlimbs. Radiologic examination identified generalized osteopenia and shortening of the limb long bones. Necropsy showed osteochondrodysplastic limbs and dilatation of the heart right ventricle. On histological examination, the physeal cartilages were characterized by multifocal mild to moderate loss of the normal columnar arrangement of chondrocytes. Osteopenia also was observed. Genetic analysis identified a missense variant in TGDS and a splice-site variant in LAMA4, both of which were homozygous in the 2 cases. Parents were heterozygous and allele frequency in the Chianina population for the TGDS variant was 5% and for the LAMA4 variant was 2%. Genetic findings identified 2 potentially pathogenic alleles in TGDS and LAMA4, but no clear mode of inheritance could be determined.

Predictors of dental arch abnormalities in children with primary dentition (part two)

Relevance. Assessing the influence of risk factors during the primary dentition period on the development of dental arch abnormalities is essential for planning preventive and therapeutic interventions, as well as encouraging compliance with these measures. The first part of this article focused on identifying predictors of malocclusion, while this second part aims to determine the predictors of dental arch abnormalities.Purpose. To identify prognostic factors (predictors) of dental arch abnormalities in children during the primary dentition period.Materials and methods. This study presents the results of a retrospective analysis of the oral health status of 123 children (55 boys and 68 girls). The initial examination took place when the children were between 4.0 and 5.5 years old (mean age 5.1 ± 0.6 years), with follow-up examinations conducted between the ages of 6.0 and 10.5 years (mean age 8.7 ± 1.3 years). The relationship between risk factors in primary dentition and the development of dental arch abnormalities in early mixed dentition was evaluated using Pearson's chi-squared test (χ2) and Welch's t-test (V). For each pair of "primary dentition risk factor – dental arch abnormality in early mixed dentition," the odds ratio with a 95% confidence interval was calculated.Results. Differentiating predisposing factors and analyzing the effects of their various combinations allowed the identification of predictor clusters in primary dentition, where the probability of developing dental arch abnormalities in mixed dentition exceeds 95%. For upper arch abnormalities, the cluster includes infantile swallowing and early extraction of deciduous molars in the lower jaw (χ2 =19.67, V=0.50); for lower arch abnormalities, the cluster consists of infantile swallowing, "lazy" chewing, sucking habits, and a deep bite in the anterior region (χ2 =16.58, V=0.67); For lower incisor crowding, the cluster includes "lazy" chewing, infantile swallowing, and a deep bite in the anterior region (χ2 =17.54, V=0.63); for diastema in the upper arch, the cluster includes an abnormal labial frenum and interdental tongue position (χ2 =19.16, V=0.49); for maxillary anterior spacing, early extraction of deciduous canines in the upper jaw is the primary factor (χ2 =16.23, V=0.46).Conclusion. Identifying and addressing predictors of dental arch abnormalities in children during the primary dentition period can significantly reduce the risk of developing pathological conditions during the mixed dentition period.

Comparison of triclosan and triclocarban in triggering immunotoxicity in larval zebrafish

As two efficient broad-spectrum sterilizing agents, triclosan (TCS) and triclocarban (TCC) are widely used, especially during the COVID-19 pandemic. The health risks caused by secondary pollution of TCS and TCC have aroused wide concern. Because of the similar mother nucleus structure and high lipophilicity, it remains unknown about the differences in the effect and mechanism of the toxicity (especially immunotoxicity) between TCS and TCC in organisms in the environment. In this study, we used zebrafish as a model to compare the immunotoxicity and mechanisms between the two pollutants at the same exposure concentration (0.6 µmol/L). The results showed that both TCS and TCC led to a hatching rate below 60% at the time point of 72 hours post fertilization (hpf) and the mortality rates of 40% and 50% at 120 hpf in larval zebrafish, respectively. The zebrafish exposed to TCS and TCC displayed malformations, such as shortened body, swimming sac closure, pericardial edema, yolk cyst deposition, and absorption disorder. Moreover, the developmental abnormalities caused by TCC were significantly severer than those caused by TCS. TCS exposure increased the proliferation rate of innate immune cells to 20% and decreased the number of mature T cells by 35%, while TCC exposure inhibited the differentiation of both innate immune cells and T cells, with the inhibition rates of 25% and 60%, respectively. The results of real-time quantitative PCR (RT-qPCR) and ELISA showed that TCS and TCC exposure up-regulated the expression levels of il-1β, il-6, and tnf-α, while il-10 and IgM exhibited opposite expression patterns. Additionally, both compounds slightly decreased C3 expression. The Pearson correlation analysis showed that the developmental toxicity induced by TCS and TCC had positive and negative correlations with the differentiation of immune cells, respectively. However, the toxicity induced by either TCS or TCC was positively correlated with the expression of pro-inflammatory cytokines. GO function and KEGG pathway enrichment analyses demonstrated that the target molecules of TCS and TCC were enriched in different signaling pathways, and the key network hub genes and the enriched regulatory pathways differed between TCS and TCC. The findings provide compelling evidence that TCS and TCC adopt different mechanisms in triggering immunotoxicity and offer a theoretical reference for the recognition, warning, and management of TCS and TCC-induced health risks.

Chitin Deacetylase 1 Gene as an Optimal RNAi-Based Target for Controlling the Tomato Leaf Miner Tuta absoluta

Chitin is a critical component of both the exoskeleton and internal structures of insects, which can protect insects from mechanical damage, dehydration and pathogen infection, and plays a significant role in the molting process. Chitin deacetylases (CDAs), key enzymes involved in chitin metabolism, are widely distributed among arthropods and microorganisms. In this study, we identified a CDA gene, TaCDA1, in the invasive insect species Tuta absoluta (Meyrick). Sequence analysis demonstrated a high degree of similarity to CDAs in other insects, revealing the presence of three conserved domains. Quantitative analysis showed that the TaCDA1 gene exhibited peak expression during the pupal stage, particularly within the epidermis. The suppression of TaCDA1 expression through RNA interference in T. absoluta pupae significantly impacted the expression of genes associated with chitin metabolism, increasing mortality and developmental abnormalities during the pupa–adult transition and reducing the pupal weight. Furthermore, soaking gene-specific dsRNA resulted in elevated mortality rates during the larva–pupa transition, causing the inability to form new cuticles or undergo ecdysis, as confirmed by subsequent histological observations. The oral administration of dsTaCDA1 + sucrose solution did not significantly impact NtCDA1 expression or the mortality rate compared to the dsGFP + sucrose solution control in the non-target insect Nesidiocoris tenuis. This study demonstrated that TaCDA1 is a potential and safe target for pest control of T. absoluta.

Drosophila ring chromosomes interact with sisters and homologs to produce anaphase bridges in mitosis.

Ring chromosomes are known in many eukaryotic organisms, including humans. They are typically associated with a variety of maladies, including abnormal development and lethality. Underlying these phenotypes are anaphase chromatin bridges that can lead to chromosome loss, nondisjunction and breakage. By cytological examination of ring chromosomes in Drosophila melanogaster we identified five causes for anaphase bridges produced by ring chromosomes. Catenation of sister chromatids appears to be the most common cause and these bridges frequently resolve during anaphase, presumably by the action of topoisomerase II. Sister chromatid exchange and chromosome breakage followed by sister chromatid union also produce anaphase bridges. Mitotic recombination with the homolog was rare, but was another route to generation of anaphase bridges. Most surprising, was the discovery of homolog capture, where the ring chromosome was connected to its linear homolog in anaphase. We hypothesize that this is a remnant of mitotic pairing and that the linear chromosome is connected to the ring by multiple wraps produced through the action of topoisomerase II during establishment of homolog pairing. In support, we showed that in a ring/ring homozygote the two rings are frequently catenated in mitotic metaphase, a configuration that requires breaking and rejoining of at least one chromosome.

Effective technology of disinfecting ozonation of hatching chicken eggs

Relevance. The positive aspects of disinfecting ozonation of hatching eggs allow us to expect successful application of the method not only in large but also in small-scale farms, as well as for experimental and industrial purposes in laboratories and bio-enterprises related to the technological process based on the incubation of a small number of eggs. This determines the importance of expanding the range of ozonizers due to numerous portable devices. There are no clear recommendations for these devices and this causes the need to search for the most effective and harmless modes for the embryo, and schemes of disinfecting ozonation.Methods. The study used fertilized chicken eggs “Hysex Brown” and a portable ozonizer “OZON-OviV”. Ozone concentration 2.0 mg / l. Eggs were treated in a specially made chamber. Technology-1: twice for 30 minutes before incubation and on the 3rd day of incubation. Technology-2: three times for 30 minutes before incubation, on days 3 and 5. The range of studies included: assessment of total microbial contamination (densitometry); identification of microorganisms (MALDI-TOF-spectrometry); biological control of incubation (fertility, hatchability, mortality, developmental abnormalities); assessment of the adequacy of the internal organs (MicroCT); embryo morphometry (weight, length, chest circumference) and calculation of development proportionality indices; histological assessment of the liver.Results. The total ozone concentration during treatment by two methods was 240 mg/l and 360 mg/l, respectively. The disinfecting efficiency of ozonation has been proven, providing a decrease in the level of total microbial contamination by 30% and 40% with double and triple treatment. A tendency to maintain a low total microbial contamination, compared to intact eggs, up to 14 days of incubation has been revealed. The dynamics of the microbial landscape indicate the bacteriostatic effect of ozone in the concentrations used on a wide range of microorganisms. Microtomographic and histological methods confirmed the harmlessness of the technologies used. Along with the more pronounced antibacterial effect of technology-2, the presence of a stimulating effect on the body of the developing embryo was revealed, which determines the preference for its choice.

Cardiograms and Electrography Are Crucial Diagnostic Procedures In Pregnant Women

Certain examinations place significant emphasis on the gestation period. Strictly passing every examination is of utmost importance for every woman, as failure to do so may result in the birth of children with various deformed and incapacitating disorders. The diagnostic procedures of cardiotomy, electrocardiography, ultrasonography, doppler ultrasonography, and phenocardiography are highly significant for pregnant women. Every expectant mother should undertake these tests comprehensively. The duration of pregnancy and developmental abnormalities like as hypotrophy, hydrocephaly, and microcephaly can be determined using these techniques. Furthermore, these techniques facilitate the surveillance of the foetal placenta and the estimation of the water content, particularly in cases of multiple pregnancies. Sonography examination is safe for both the pregnant woman and the developing foetus. Today, X-ray diagnostics is seldom employed. However, this approach strictly relies on the use of the vagina as the sole means of assessing the state and abnormalities of the foetus, in the absence of ultrasound equipment.

Neurodevelopmental outcome in children between one and five years after persistent pulmonary hypertension of term and near-term newborns

BackgroundPersistent pulmonary hypertension of the newborn (PPHN) is a serious condition that affects 1–2 per 1,000 newborns. Scientific data report the existence of neurological developmental abnormalities between 10 and 30%, but the description of these disorders linked with this situation of cerebral hypoxia and haemodynamic failure remains poorly documented.ObjectiveThe main goal of this study was to describe the prevalence of neuro-psychomotor developmental disorders in children aged between one and five years old who have been hospitalised at birth in a neonatal intensive care unit for the management of PPHN.MethodsAll of the newborns ≥34 weeks of gestational age (WGA) with PPHN, treated with inhaled nitric oxide in our neonatal intensive care unit between January 2015 and December 2019 were retrospectively enrolled. An ASQ-3 standardised questionnaire, adapted to the appropriate age (12, 24, 36, 48 and 60 months) was performed by the parents.ResultsFifty-five children (81% of answers) with a median age of 36 months (11–68), whose real age was close to the one of the questionnaire (12, 24, 36, 48 and 60 months), have been included in this study. There was 47% of pathological score [borderline: less than 1 standard deviation (SD) or suspect: less than 2SD] in at least one of the five studied domains, mainly in communication (25%) and individual and social skills (22%), despite a high overall score of 250 [220; 285] out of 300 that improved with age.ConclusionThis study showed a significant prevalence of neuro-psychomotor developmental disorders which justifies making more accessible a prolonged and adapted follow-up for early and multidisciplinary screening and management of these children with PPHN history. Larger cohorts are needed to better explore long term outcome of these vulnerable term neonates.

Uncovering the Embryonic Origins of Duchenne Muscular Dystrophy.

Duchenne muscular dystrophy (DMD) is a severe degenerative muscle disease caused by mutations in the DMD gene, which encodes dystrophin. Despite its initial description in the late 19th century by French neurologist Guillaume Duchenne de Boulogne, and identification of causal DMD genetic mutations in the 1980s, therapeutics remain challenging. The current standard of care is corticosteroid treatment, which delays the progression of muscle dysfunction but is associated with significant adverse effects. Emerging therapeutic approaches, including AAV-mediated gene transfer, CRISPR gene editing, and small molecule interventions, are under development but face considerable obstacles. Although DMD is viewed as a progressive muscle disease, muscle damage and abnormal molecular signatures are already evident during fetal myogenesis. This early onset of pathology suggests that the limited success of current therapies may partly be due to their administration after aberrant embryonic myogenesis has occurred in the absence of dystrophin. Consequently, identifying optimal therapeutic strategies and intervention windows for DMD may depend on a better understanding of the earliest DMD disease mechanisms. As newer techniques are applied, the field is gaining increasingly detailed insights into the early muscle developmental abnormalities in DMD. A comprehensive understanding of the initial events in DMD pathogenesis and progression will facilitate the generation and testing of effective therapeutic interventions.

Systemic effect of the oncological process on the dog’s body

The article presents the results of an analysis of a non-randomized study of the prevalence of paraneoplastic syndromes in primary spontaneous malignant neoplasms in dogs. The study was conducted in 2022–2024 on the basis of the Department of Diseases of Small Domestic, Laboratory and Exotic Animals and the Research Laboratory of Oncology, Ophthalmology and Animal Biochemistry of the Russian Biotechnological University “ROSBIOTECH”. The object of the study were 948 dogs with primary spontaneous malignant neoplasms confirmed by morphological methods.The aim of the work was to study the frequency of paraneoplastic syndromes in dogs with neoplasia.The diagnosis of paraneoplastic syndrome was made by excluding other possible etiological factors for the development of clinical and laboratory abnormalities identified during a comprehensive examination of dogs. As a result of the study, the incidence of paraneoplasia was 54.54% of cases. Hematological (44.41%) and gastrointestinal (37.13%) paraneoplastic syndromes were most often recorded, less often endocrine (3.27%) and ophthalmological (3.38%), extremely rarely renal (1.27%), osteoarticular (0.42%) and dermatological (0.42%). The main hematological manifestations of the systemic effect of the oncological process were: anemia (12.87%), neutrophilic leukocytosis (12.76%) and thrombocytopenia (8.86%). Gastrointestinal paraneoplastic syndrome in most cases was manifested by a decrease in body weight (33.65%) and hyporexia (18.04%), inflammatory and erosive ulcerative lesions of the gastrointestinal tract (7.59%). During the study, a case of a nonspecific systemic reaction to a malignant tumor in the form of fever in the absence of infectious and inflammatory processes (0.11%) was registered.

Trabecular Meshwork Abnormalities in a Model of Congenital Glaucoma Due to LTBP2 Mutation.

To characterize early trabecular meshwork (TM) morphologic abnormalities in a feline model of human primary congenital glaucoma (PCG) caused by mutation in LTBP2. Eyes from 41 cats, including 19 normal and 22 homozygous for LTBP2 mutation, across various postnatal stages (birth, 2 weeks, 5 weeks, and 12 weeks) were paraformaldehyde fixed, anterior segments dissected, post-fixed in glutaraldehyde, osmicated, and processed and sectioned for transmission electron microscopy. Cell morphology, nuclear shape, and intertrabecular space (ITS) were quantitatively assessed, and the structure of the fibrillar extracellular matrix in the TM was systematically evaluated. The earliest differences in TM morphology between PCG and normal cats were identified at 2 weeks postnatally. Elastic fibers in the TM were discontinuous and disorganized (P = 0.0122), and by 5 weeks of age PCG cats presented significantly less ITS (P = 0.0076) and morphologically rounder TM cells than normal cats (P = 0.0293). By 12 weeks of age, the ITS was further collapsed (P < 0.0001), and the TM cells were morphologically elongated and attenuated in PCG compared to controls (P = 0.0028). In this feline model of PCG due to LTBP2 mutation, development of ultrastructural TM extracellular matrix abnormalities are first observed by 2 weeks and cellular abnormalities by 5 weeks of age. By 12 weeks of age, when intraocular pressure becomes significantly elevated, the TM morphologic abnormalities are already well established. These findings suggest that the postnatal period between 0 and 5 weeks of age is critical for TM and PCG development and progression in cats.

Clinical and immunologic abnormalities in COVID-19

Objective: to find approaches to improve diagnostics of the debut of rheumatic manifestations, associated with COVID-19.Material and methods. Data from 1000 patients from the COVID-19 registry were included in the prospective cohort study. In all patients, the diagnosis of COVID-19 was confirmed by polymerase chain reaction. Of these patients, 380 (41.8% men and 58.2% women, mean age 47.0±2.5 years) had rheumatic manifestations. Patients were examined using routine clinical methods. Immunological markers of rheumatic diseases were determined, including antibodies against cyclic citrullinated peptide, rheumatoid factor, antiphospholipid antibodies and antinuclear factor (ANF), and an immunoblot for antinuclear antibodies was performed if ANF titer was &gt;1:160.Results and discussion. Patients had the following rheumatic manifestations: arthralgias (in 342), myalgias (in 23), skin rashes (in 15). ANF titers &gt;1:160 were found in 57.6% of patients. No reliable data indicating the development of an antiphospholipid syndrome were found in the study group. Lupus anticoagulant was detected in 5.7% of cases, antibodies against β2-glycoprotein in 5.7%, antibodies against cardiolipin in 3.8%. High ANF titers were found in 63.9% of patients with arthralgia. Gender-specific differences were found when analyzing the correlation between ANF titers and rheumatic manifestations: in men, high ANF tires were associated with myalgias, and in women with arthralgias. The presence of rheumatic manifestations depended directly on the severity of the disease. A correlation between arthralgia and leucopenia was also found – leucocyte count &lt; 3,9 ‧109 /L was a predictor of arthralgias. The sensitivity and specificity of the model were 99.3 and 91.2%, respectively.Conclusion. The results suggest that COVID-19 can provoke the development of immunological abnormalities that may subsequently lead to the development of an autoimmune diseases (AID). The optimal approach to prevention and early detection of AID in patients with coronavirus infection caused by SARS-CoV-2 is to monitor laboratory parameters – leukocyte count and CRP level. If rheumatic manifestations are present, the use of immunological and imaging examinations is also recommended.

Toxicity of nuclear-localized GFP in reporter mice

Various techniques using fluorescent reporter probes have been developed, such as GFP transgenic mouse lines that are used to detect spatial–temporal expression levels of genes. Although GFP expression is largely considered non-toxic, recent reports have indicated that under certain conditions GFP can display cellular toxicity. We hereby report the nuclear toxicity of H2B-GFP using a K14 specific Tet-on reporter mouse system. Using this system, GFP accumulates in the nucleus of all K14 expressing cells, such as the ocular surface epithelia and ocular adnexa. Expression of high levels of nuclear GFP during embryonic stages led to an eye open-at-birth (EOB) phenotype and abnormal ocular adnexa development and during adult and aging stages showed notable toxicity to ocular tissues. Other tissues, such as skin, also presented multiple defects associated with H2B-GFP expression. This toxicity was found to be concentration dependent, with homozygous mice presenting extremely high toxicity, while heterozygous mice presented limited toxicity. Upon induction, the accumulation of H2B-GFP in the nucleus of homozygous mice led to apoptosis within 2 weeks. This study therefore shows that although the use of nuclear GFP reporter mice is a valuable tool, at high levels, nuclear GFP can be toxic, leading to cell death and affecting tissue function.