Expression pattern of germ cell markers in cryptorchid stallion testes.

Cryptorchidism affects spermatogenesis and testis development, often resulting in stallion subfertility/infertility. This study aims to identify the specific germ cells impacted by cryptorchism in stallions. In a previous study, we found that PGP9.5 and VASA are molecular markers expressed in different germ cells within stallions. Herein, we assessed the heat stress-induced response of spermatogonial stem cells (SSCs) in the seminiferous tubules (ST) of cryptorchid stallion testes (CST) and normal stallion testes (NST). This goal was accomplished by comparing PGP9.5 and VASA expression patterns through reverse transcription quantitative PCR and immunofluorescence assays. We also compared the cross-sectional ST area between groups. Six post-pubertal Thoroughbred unilateral cryptorchid stallions were used. The relative abundance of the mRNA transcripts of PGP9.5 and VASA was significantly upregulated in the NST group than in the CST group. Additionally, the cross-sectional ST area and localization of PGP9.5 and VASA in germ cells were significantly higher in the NST group than in the CST group. Regarding Leydig cells, PGP9.5 staining was observed in both groups. Spermatogonia, primary spermatocytes and secondary spermatocytes were immunostained with VASA in the NST group, while immunostaining was only observed in spermatogonia in the CST group. These results indicate long-term exposure to heat stress conditions, such as cryptorchidism, directly impacts germ cell proliferation and differentiation, leading to impaired spermatogenesis and compromised fertility in stallions.

Challenging behaviour and epilepsy in patients with intellectual disability: frequency and mutual association.

Comorbid epilepsy and challenging behaviours is quiet common in patients with ID (intellectual disability). This study aims to determine the frequency and mutual association between epilepsy and challenging behaviours. In this cross-sectional analytical study, 252 patients were enrolled through convenient sampling technique. Comorbid epilepsy and CB (challenging behaviour) were seen in 111 (44.6%) and 116 (46.6%) patients, respectively. Epilepsy and severity of intellectual disability (ID) are statistically and significantly associated with challenging behaviour. This study concluded that comorbid epilepsy is more common among people with ID as compared to the general population. The clinical variables, i.e. comorbid epilepsy and severity of ID have statistically significant association with the CB.

Investigating vertical transmission and maternofoetal outcomes in pregnant women with COVID-19: a case-control study from Pakistan.

Objectives: To estimate the frequency of severe acute respiratory syndrome coronavirus-2 among pregnant women, the impact in terms of obstetrical and clinical outcomes and vertical transmission to the neonates. METHODS: The prospective, case-control study was conducted at Zainab Panjwani Memorial Hospital, Karachi, from March to December 2021, and comprised pregnant women regardless of gestational age who exhibited symptoms or had a suspicion of exposure to any confirmed coronavirus disease-2019 individual. They were screened for severe acute respiratory syndrome coronavirus-2 infection using polymerase chain reaction or serology. Those who tested negative were designated as control group A, while who had a positive serology result along with a negative polymerase chain reaction were taken as recovered case group B1, and those who tested positive for polymerase chain reaction were called the positive case group B2. Groups B1 and B2 were followed up till delivery. The clinical presentation of coronavirus disease-2019 infection in pregnancy and its obstetrical and neonatal outcomes was assessed. Products of conception were tested for the detection of the severe acute respiratory syndrome coronavirus-2 genome. The viral genome from group B2 cases was sequenced to confirm vertical transmission. Data was analysed using GraphPad Prism V8. RESULTS: Of the 139 pregnant women, 74(53.2%) were in group A with mean age 25.87±6.90 years, 49(35.3%) were in group B1 with mean age 25.53±7.02 years, and 16(11.5%) were in group B2 with mean age 27.12±5.03 years. The gestational age at which termination of pregnancy occurred was 38.3±1.26 weeks in group B1 and 38.3±1.85 weeks for group B2. There were 96 neonates across the 3 groups. Of the 11(11.45%) neonates in group B2, 1(9.09%) had postnatal transmission of severe acute respiratory syndrome coronavirus-2 and this mother-neonate case was taken as the Indexed case. The severe acute respiratory syndrome coronavirus-2 genome isolated from the neonate showed similar mutations as the viral strain infecting the mother. Conclusion: The risk of vertical transmission was found to be low. The severe acute respiratory syndrome coronavirus-2 genome was the same for both the mother and the neonate.

Changes in characteristics of spermatogonial stem cells in response to heat stress in stallions.

Spermatogonial stem cells (SSCs) are essential for the maintenance of male fertility and survival of species. Environmental conditions, notably heat stress, have been identified as important causes of male infertility and have a negative impact on SSCs. Animals with cryptorchid testes (CT) are optimal models for the study of long-term heat stress-related changes in germ cells. The effect of heat stress on germ cells differs depending on the spermatogenesis stage. Thus, verifying whether the specific phase of spermatogenesis is dependent or independent of heat stress in stallions is important. We evaluated the heat stress-related response of SSCs by comparing the relative abundance of mRNA transcripts and expression patterns of the undifferentiated embryonic cell transcription factor 1 (UTF-1) and deleted in azoospermia-like (DAZL) in the seminiferous tubules of CT and normal testes (NT) of stallions using reverse transcription-quantitative polymerase chain reaction (RT-qPCR), immunofluorescence, and western blotting. We also analyzed the relative abundance of mRNA of different proliferative markers, including minichromosome maintenance 2 (MCM2), marker of proliferation Ki-67 (MKI-67), and proliferating cell nuclear antigen (PCNA). Testicular tissues from four Thoroughbred unilateral cryptorchid postpubertal stallions were used in this study during the breeding season. The relative abundance of the mRNA transcripts of UTF-1 and MCM2 was significantly upregulated in the CT group than that of those in the NT group. In contrast, the relative abundance of the mRNA transcripts of DAZL was significantly downregulated in the CT group than that of those in the NT group. Western blot quantification showed that the relative intensity of UTF-1 protein bands was significantly higher, while that of DAZL protein bands was significantly lower in the CT group than in the NT group. Immunofluorescence studies showed that the number of germ cells immunostained with UTF-1 was significantly higher while immunostained with DAZL was significantly lower in the CT group than that in the NT group. The higher expression level of UTF-1 in the CT group shows that undifferentiated SSCs are not affected by long-term exposure to heat stress. These results also indicate that germ cells after differentiation phase are directly affected by heat-stress conditions, such as cryptorchidism, in stallions.

Predisposing deleterious variants in the cancer-associated human kinases in the global populations.

Human kinases play essential and diverse roles in the cellular activities of maintaining homeostasis and growth. Genetic mutations in the genes encoding the kinases (or phosphotransferases) have been linked with various types of cancers. In this study, we cataloged mutations in 500 kinases genes in >65,000 individuals of global populations from the Human Genetic Diversity Project (HGDP) and ExAC databases, and assessed their potentially deleterious impact by using the in silico tools SIFT, Polyphen2, and CADD. The analysis highlighted 35 deleterious non-synonymous SNVs in the ExAC and 5 SNVs in the HGDP project. Notably, a higher number of deleterious mutations was observed in the Non-Finnish Europeans (26 SNVs), followed by the Africans (14 SNVs), East Asians (13 SNVs), and South Asians (12 SNVs). The gene set enrichment analysis highlighted NTRK1 and FGFR3 being most significantly enriched among the kinases. The gene expression analysis revealed over-expression of NTRK1 in liver cancer, whereas, FGFR3 was found over-expressed in lung, breast, and liver cancers compared to their expression in the respective normal tissues. Also, 13 potential drugs were identified that target the NTRK1 protein, whereas 6 potential drugs for the FGFR3 target were identified. Taken together, the study provides a framework for exploring the predisposing germline mutations in kinases to suggest the underlying pathogenic mechanisms in cancers. The potential drugs are also suggested for personalized cancer management.

Lie symmetry analysis and solitary wave solution of biofilm model Allen-Cahn.

The investigation presented in this study delves into the analysis of Lie symmetries for the bistable Allen-Cahn (BAC) equation with a quartic potential, specifically applied to the biofilm model. By employing the Lie symmetry method, we have acquired the Lie infinitesimal generators for the considered model. Using a transformation method, the nonlinear partial differential equations (NPDEs) are converted into various nonlinear ordinary differential equations (NLODEs), providing the numerous closed-form solitary wave solutions. The obtained solutions manifest in various forms including dark, bright, kink, anti-kink, and periodic types using diverse strategies. To enhance the physical interpretation, the study presents 3D, 2D, and contour plots of the acquired solutions. Every graph's wave-like structure contains information about the structural behaviour of the bacteria that build biofilms on surfaces where rectangles have different densities. This analysis enhances comprehension of the complex dynamics present in areas like fluid dynamics, fiber optics, biology, ocean physics, coastal engineering, and nonlinear complex physical systems.

Intelligent prediction modeling for flexural capacity of FRP-strengthened reinforced concrete beams using machine learning algorithms.

Fiber-reinforced polymers (FRP) are widely utilized to improve the efficiency and durability of concrete structures, either through external bonding or internal reinforcement. However, the response of FRP-strengthened reinforced concrete (RC) members, both in field applications and experimental settings, often deviates from the estimation based on existing code provisions. This discrepancy can be attributed to the limitations of code provisions in fully capturing the nature of FRP-strengthened RC members. Accordingly, machine learning methods, including gene expression programming (GEP) and multi-expression programming (MEP), were utilized in this study to predict the flexural capacity of the FRP-strengthened RC beam. To develop data-driven estimation models, an extensive collection of experimental data on FRP-strengthened RC beams was compiled from the experimental studies. For the assessment of the accuracy of developed models, various statistical indicators were utilized. The machine learning (ML) based models were compared with empirical and conventional linear regression models to substantiate their superiority, providing evidence of enhanced performance. The GEP model demonstrated outstanding predictive performance with a correlation coefficient (R) of 0.98 for both the training and validation phases, accompanied by minimal mean absolute errors (MAE) of 4.08 and 5.39, respectively. In contrast, the MEP model achieved a slightly lower accuracy, with an R of 0.96 in both the training and validation phases. Moreover, the ML-based models exhibited notably superior performances compared to the empirical models. Hence, the ML-based models presented in this study demonstrated promising prospects for practical implementation in engineering applications. Moreover, the SHapley Additive exPlanation (SHAP) method was used to interpret the feature's importance and influence on the flexural capacity. It was observed that beam width, section effective depth, and the tensile longitudinal bars reinforcement ratio significantly contribute to the prediction of the flexural capacity of the FRP-strengthened reinforced concrete beam.

Exploration of newly synthesized amantadine-thiourea conjugates for their DNA binding, anti-elastase, and anti-glioma potentials.

Three newly synthesized amantadine thiourea conjugates namely MS-1 N-(((3 s,5 s,7 s)-adamantan-1-yl)carbamothioyl)benzamide, MS-2 N-(((3 s,5 s,7 s)-adamantan-1-yl)carbamothioyl)-4-methylbenzamide and MS-3 N-((3 s,5 s,7 s)-adamantan-1-ylcarbamothioyl)-4-chlorobenzamide were investigated for their structures, bindings (DNA/ elastase), and for their impact on healthy and cancerous cells. Theoretical (DFT/docking) and experimental {UV-visible (UV-), fluorescence (Flu-), and cyclic voltammetry (CV)} studies indicated binding interactions of each conjugate with DNA and elastase enzyme. Theoretically and experimentally calculated binding parameters for conjugate - DNA interaction revealed MS-3 - DNA to have most significant binding with comparatively greater values of binding parameters {(Kb/M-1: docking, 3.8 × 105; UV-, 5.95 × 103; Flu-,1.55 × 105; CV, 1.52 × 104), (∆G/ kJmol-1: docking, -32.09; UV-, -22.40; Flu-,-30.81; CV, -24.82)}. The docked structures, greater bindings site size values (n), and the trend in DNA viscosity changes in the presence of each conjugate concentration confirmed a mixed binding mode of interaction among them. Conjugate - elastase binding by docking agreed with the experimental anti-elastase findings. Cytotoxicity studies of each tested conjugate demonstrated greater cytotoxicity for cancerous (MG-U87) cells in comparison to control, while for the normal (HEK-293) cells the cytotoxicity was found comparatively low. Overall exploration suggested that MS-3 is the most effective candidate for DNA binding, anti-elastase, and for anti-glioma activities.

Noval soliton solution, sensitivity and stability analysis to the fractional gKdV-ZK equation.

This work examines the fractional generalized Korteweg-de-Vries-Zakharov-Kuznetsov equation (gKdV-ZKe) by utilizing three well-known analytical methods, the modified [Formula: see text]-expansion method, [Formula: see text]-expansion method and the Kudryashov method. The gKdV-ZK equation is a nonlinear model describing the influence of magnetic field on weak ion-acoustic waves in plasma made up of cool and hot electrons. The kink, singular, anti-kink, periodic, and bright soliton solutions are observed. The effect of the fractional parameter on wave shapes have been analyzed by displaying various graphs for fractional-order values of [Formula: see text]. In addition, we utilize the Hamiltonian property to observe the stability of the attained solution and Galilean transformation for sensitivity analysis. The suggested methods can also be utilized to evaluate the nonlinear models that are being developed in a variety of scientific and technological fields, such as plasma physics. Findings show the effectiveness simplicity, and generalizability of the chosen computational approach, even when applied to complex models.

Non-invasive detection of mental fatigue in construction equipment operators through geometric measurements of facial features.

INTRODUCTION: Prolonged operation of construction equipment could lead to mental fatigue, which can increase the chances of human error-related accidents as well as operators' ill-health. The objective detection of operators' mental fatigue is crucial for reducing accident risk and ensuring operator health. Electroencephalography, photoplethysmography, electrodermal activity, and eye-tracking technology have been used to mitigate this issue. These technologies are invasive and wearable sensors that can cause irritation and discomfort. Geometric measurements of facial features can serve as a noninvasive alternative approach. Its application in detecting mental fatigue of construction equipment operators has not been reported in the literature. Although the application of facial features has been widespread in other domains, such as drivers and other occupation scenarios, their ecological validity for construction excavator operators remains a knowledge gap. METHOD: This study proposed employing geometric measurements of facial features to detect mental fatigue in construction equipment operators' facial features. In this study, seventeen operators performed excavation operations. Mental fatigue was labeled subjectively and objectively using NASA-TLX scores and EDA values. Based on geometric measurements, facial features (eyebrow, mouth outer, mouth corners, head motion, eye area, and face area) were extracted. RESULTS: The results showed that there was significant difference in the measured metrics for high fatigue compared to low fatigue. Specifically, the most noteworthy variation was for the eye and face area metrics, with mean differences of 45.88% and 26.9%, respectively. CONCLUSIONS: The findings showed that geometrical measurements of facial features are a useful, noninvasive approach for detecting the mental fatigue of construction equipment operators.

Novel acrylamide-based baits for effective control of subterranean termites (Blattodea: Rhinotermitidae).

BACKGROUND: Acrylamide-based bait has super water absorption making it highly attractive to subterranean termites that are lured by wood with high water content. This study investigated the control efficiency of these baits on subterranean termites. In particular, we evaluated the water-absorption capacity, attractiveness to subterranean termites, and control efficiency of these baits on subterranean termites through wooden blocks (Populus deltoides and three types of particleboards). RESULTS: The results indicated a substantial water absorption capacity of acrylamide (70.6%; control: 14.8%) and a strong attraction for feeding subterranean termites (P. deltoides: 198 highest; 81 lowest subterranean termites individuals; combination of neem leaves and walnut shells: 168 highest; 36 lowest subterranean termites individuals). When acrylamide was combined with boric acid at the highest concentration, it resulted in the lowest wood consumption rates (P. deltoides: 24.1%; control: 63.8%, combination of neem leaves and walnut shells: 32.5%; control: 62.1%). CONCLUSIONS: In conclusion, this research supports the commercial viability of employing innovative acrylamide-based toxic baits and particleboards for subterranean termite management. © 2024 Society of Chemical Industry.

Molecular identification, characterization, and expression analysis of a serine protease inhibitor gene from cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae) / Identificação molecular, caracterização e análise de expressão de um gene inibidor de serina protease da lagarta Helicoverpa armigera (Lepidoptera: noctuidae) na cultura de algodão

Abstract Serine protease inhibitors (serpins), a superfamily of protease inhibitors, are known to be involved in several physiological processes, such as development, metamorphosis, and innate immunity. In our study, a full-length serpin cDNA, designated Haserpin1, was isolated from the cotton bollworm Helicoverpa armigera. The cDNA sequence of Haserpin1 is 1176 nt long, with an open reading frame encoding 391 amino acids; there is one exon and no intron. The predicted molecular weight of Haserpin1 is 43.53 kDa, with an isoelectric point of 4.98. InterProScan was employed for Haserpin1 functional characterization, which revealed that Haserpin1 contains highly conserved signature motifs, including a reactive center loop (RCL) with a hinge region (E341-N350), the serpin signature, (F367-F375) and a predicted P1-P1′ cleavage site (L357-S358), which are useful for identifying serpins. Transcripts of Haserpin1 were constitutively expressed in the fat body, suggesting that it is the major site for serpin synthesis. During the developmental stages, a fluctuation in the expression level of Haserpin1 was observed, with low expression detected at the 5th-instar larval stage. In contrast, relatively high expression was detected at the prepupal stage, suggesting that Haserpin1 might play a critical role at the H. armigera wandering stage. Although the detailed function of this serpin (Haserpin1) needs to be elucidated, our study provides a perspective for the functional investigation of serine protease inhibitor genes.

Resumo Sabe-se que os inibidores de serina protease (serpinas), uma superfamília de inibidores de protease, estão envolvidos em vários processos fisiológicos, como desenvolvimento, metamorfose e imunidade inata. Neste estudo, um cDNA de serpina de comprimento total, denominado Haserpin1, foi isolado da lagarta Helicoverpa armigera na cultura de algodão. A sequência de ADNc de Haserpin1 tem 1.176 nt de comprimento, com uma grelha de leitura aberta que codifica 391 aminoácidos; existe um éxon, mas nenhum íntron. O peso molecular previsto de Haserpin1 é de 43,53 kDa, com um ponto isoelétrico de 4,98. O InterProScan foi empregado para a caracterização funcional do Haserpin1, que revelou que o Haserpin1 contém motivos de assinatura altamente conservados, incluindo um loop central reativo (RCL) com uma região de dobradiça (E341-N350), a assinatura da serpina (F367-F375) e um local de clivagem previsto de P1-P1' (L357-S358), que são úteis para identificar serpinas. As transcrições de Haserpin1 foram expressas constitutivamente no corpo gordo, sugerindo que é o principal local para a síntese de serpinas. Durante os estágios de desenvolvimento, observou-se uma flutuação no nível de expressão de Haserpin1, com baixa expressão detectada no estágio larval do 5º ínstar. Por outro lado, detectou-se uma expressão relativamente alta no estágio pré-pupal, sugerindo que o Haserpin1 pode desempenhar um papel crítico no estágio errante de H. armigera. Embora a função detalhada dessa serpina (Haserpin1) precise ser elucidada, este estudo fornece uma perspectiva para a investigação funcional dos genes inibidores da serina protease.

Molecular identification, characterization, and expression analysis of a serine protease inhibitor gene from cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae)

Abstract Serine protease inhibitors (serpins), a superfamily of protease inhibitors, are known to be involved in several physiological processes, such as development, metamorphosis, and innate immunity. In our study, a full-length serpin cDNA, designated Haserpin1, was isolated from the cotton bollworm Helicoverpa armigera. The cDNA sequence of Haserpin1 is 1176 nt long, with an open reading frame encoding 391 amino acids; there is one exon and no intron. The predicted molecular weight of Haserpin1 is 43.53 kDa, with an isoelectric point of 4.98. InterProScan was employed for Haserpin1 functional characterization, which revealed that Haserpin1 contains highly conserved signature motifs, including a reactive center loop (RCL) with a hinge region (E341N350), the serpin signature, (F367F375) and a predicted P1P1 cleavage site (L357S358), which are useful for identifying serpins. Transcripts of Haserpin1 were constitutively expressed in the fat body, suggesting that it is the major site for serpin synthesis. During the developmental stages, a fluctuation in the expression level of Haserpin1 was observed, with low expression detected at the 5th-instar larval stage. In contrast, relatively high expression was detected at the prepupal stage, suggesting that Haserpin1 might play a critical role at the H. armigera wandering stage. Although the detailed function of this serpin (Haserpin1) needs to be elucidated, our study provides a perspective for the functional investigation of serine protease inhibitor genes.

Resumo Sabe-se que os inibidores de serina protease (serpinas), uma superfamília de inibidores de protease, estão envolvidos em vários processos fisiológicos, como desenvolvimento, metamorfose e imunidade inata. Neste estudo, um cDNA de serpina de comprimento total, denominado Haserpin1, foi isolado da lagarta Helicoverpa armigera na cultura de algodão. A sequência de ADNc de Haserpin1 tem 1.176 nt de comprimento, com uma grelha de leitura aberta que codifica 391 aminoácidos; existe um éxon, mas nenhum íntron. O peso molecular previsto de Haserpin1 é de 43,53 kDa, com um ponto isoelétrico de 4,98. O InterProScan foi empregado para a caracterização funcional do Haserpin1, que revelou que o Haserpin1 contém motivos de assinatura altamente conservados, incluindo um loop central reativo (RCL) com uma região de dobradiça (E341-N350), a assinatura da serpina (F367-F375) e um local de clivagem previsto de P1-P1' (L357-S358), que são úteis para identificar serpinas. As transcrições de Haserpin1 foram expressas constitutivamente no corpo gordo, sugerindo que é o principal local para a síntese de serpinas. Durante os estágios de desenvolvimento, observou-se uma flutuação no nível de expressão de Haserpin1, com baixa expressão detectada no estágio larval do 5º ínstar. Por outro lado, detectou-se uma expressão relativamente alta no estágio pré-pupal, sugerindo que o Haserpin1 pode desempenhar um papel crítico no estágio errante de H. armigera. Embora a função detalhada dessa serpina (Haserpin1) precise ser elucidada, este estudo fornece uma perspectiva para a investigação funcional dos genes inibidores da serina protease.