Neddylation inhibition affects early embryonic development by disrupting maternal-to-zygotic transition and mitochondrial function in mice.

Post-translational modifications (PTMs) are critical for early development in mice because early cleavage-stage embryos are characterized by transcriptional inactivity. Neddylation is an important ubiquitin-like PTM that regulates multiple biophysical processes. However, the exact roles of neddylation in regulating early embryonic development remain largely unknown. In the present study, we found that inhibition of neddylation by specific inhibitor MLN4924 led to severe arrest of early embryonic development. Transcriptomic analysis showed that neddylation inhibition changed the expression of 3959 genes at the 2-cell stage. Importantly, neddylation inhibition blocked zygotic genome activation and maternal mRNA degradation, thus disrupting the maternal-to-zygotic transition. Moreover, inhibition of neddylation induced mitochondrial dysfunction including aberrant mitochondrial distribution, decreased mitochondrial membrane potential, and reduced ATP content. Further analysis showed that inhibition of neddylation resulted in the accumulation of reactive oxygen species and superoxide anion, thereby resulting in oxidative stress and severe DNA damage at the 2-cell stage. Overall, this study demonstrates that neddylation is vital for early embryonic development in mice. Our findings suggest that proper neddylation regulation is essential for the timely inter-stage transition during early embryonic development.

Integrating mid-infrared spectroscopy, machine learning, and graphical bias correction for fatty acid prediction in water buffalo milk.

BACKGROUND: Buffalo milk, constituting 15% of global production, has higher fatty acids content than Holstein milk. Fourier-transform mid-infrared (FT-MIR) spectroscopy is widely used for dairy analysis, but its application to buffalo milk, with larger fat globules, remains understudied. The ultimate goal of this study is to develop machine learning models based on FT-MIR spectroscopy for predicting fatty acids in buffalo milk and to assess the accuracy of commercial milk analyzers. This research provides a convenient, fast, and environmentally friendly method for detecting the fatty acid composition in buffalo milk. RESULTS: We employed six machine learning algorithms to establish a detection model for 34 fatty acids in buffalo milk. The predictive models demonstrated robust capabilities for high-content fatty acids [C14:0, C15:0, C16:0, C17:0, C18:0, C18:1, saturated fatty acid (SFA), monounsaturated fatty acid (MUFA)], with errors within a 15% range. Traditional FT6000 detection methods exhibited limitations in measuring SFAs and polyunsaturated fatty acids (PUFA). Implementing a mean difference correction of 0.21 for MUFAs and applying regression equations (SFA × 1.0639 + 0.0705; PUFA × 0.5472 + 0.0047) significantly improved measurement accuracy. CONCLUSION: This study successfully developed a predictive model for fatty acids in Mediterranean buffalo milk based on FT-MIR spectroscopy. Additionally, a correction was applied to the existing measurement device, FT6000, enabling more accurate measurements of fatty acids in buffalo milk. The findings have practical implications for the food industry, offering a faster and more reliable approach to assess and monitor fatty acid composition in buffalo milk, potentially influencing product development and quality control processes. © 2024 Society of Chemical Industry.

RNA Sequencing Reveals the Inhibitory Effect of High Levels of Arachidonic Acid and Linoleic Acid on C2C12 Differentiation and Myogenic Biomarkers.

Over the past three decades, studies have shown that consuming polyunsaturated fatty acids (PUFAs) can enhance animal and human health and welfare through biological, biochemical, pathological, and pharmacological impacts. Furthermore, omega-6 plays key roles in the cardiopulmonary system, including promoting airway relaxation and inhibiting atherosclerosis and hypertension. However, findings from investigations of the effects of omega-6 fatty acids on molecular and cellular activity and discussions on their influence on biomarkers are still unclear. Therefore, the present study aimed to evaluate omega-6 fatty acids, the arachidonic acid (AA), and linoleic acid (LA) effects on C2C12 proliferation, myogenesis morphology, and relative myogenic biomarker expression through the Wnt pathway. C2C12 cells were cultured with and without 25, 50, 100, and 150 µM of LA and AA and then subjected to CCK8, Giemsa staining, RT qPCR, Western blotting, and RNA Sequencing. The CCK8 Assay results showed that 25, 50, 100, and 150 µM LA significantly decreased the viability after 72 h for 25, 50, 100, and 150 µM concentrations. Also, AA supplementation decreased cell viability after 24 h for 150 µM, 48 h for 150 µM, and 72 h for 50, 100, and 150 µM concentrations. Moreover, the LA and AA inhibitory effects noticed through Gimesa staining were morphological changes during myoblast differentiation. Both LA and AA showed inhibiting IGF1, Cola1, Col6a2, Col6a1, Itga10, Itga11, SFRP2, DAAM2, and NKD2 effects; however, the depressing effect was higher for AA compared to LA. The previous results were confirmed through Western blotting, which showed that 50 µM LA and AA significantly reduced DAAM2 and SFRP2 protein levels compared to the control. Regarding RNA sequencing results, LA and AA increased the number of differentially expressed (DE) Mt-rRNA and snoRNA; however, the numbers of lncRNA detected decreased compared to the control. Our findings demonstrate that high and moderate LA and AA concentrations reduce primary myoblast proliferation and differentiation. Also, they highlight novel biomarkers and regulatory factors to improve our understanding of how the nutrition of fatty acids can control and modulate the myogenesis and differentiation process through different biomarker families.

NADPH oxidase 2 mediates cardiac sympathetic denervation and myocyte autophagy, resulting in cardiac atrophy and dysfunction in doxorubicin-induced cardiomyopathy.

Doxorubicin has been used extensively as a potent anticancer agent, but its clinical use is limited by its cardiotoxicity. However, the underlying mechanisms remain to be fully elucidated. In this study, we tested whether NADPH oxidase 2 (Nox2) mediates cardiac sympathetic nerve terminal abnormalities and myocyte autophagy, resulting in cardiac atrophy and dysfunction in doxorubicin-induced heart failure. Nox2 knockout (KO) and wild-type (WT) mice were randomly assigned to receive a single injection of doxorubicin (15 mg/kg, i.p.) or saline. WT doxorubicin mice exhibited the decreases in survival rate, left ventricular (LV) wall thickness and LV fractional shortening and the increase in the lung wet-to-dry weight ratio 1 week after the injections. These alterations were attenuated in Nox2 KO doxorubicin mice. In WT doxorubicin mice, myocardial oxidative stress was increased, myocardial noradrenergic nerve fibers were reduced, myocardial expression of PGP9.5, GAP43, tyrosine hydroxylase and norepinephrine transporter was decreased, and these changes were prevented in Nox2 KO doxorubicin mice. Myocyte autophagy was increased and myocyte size was decreased in WT doxorubicin mice, but not in Nox2 KO doxorubicin mice. Nox2 mediates cardiac sympathetic nerve terminal abnormalities and myocyte autophagy-both of which contribute to cardiac atrophy and failure after doxorubicin treatment.

A New Method to Detect Buffalo Mastitis Using Udder Ultrasonography Based on Deep Learning Network.

Mastitis is one of the most predominant diseases with a negative impact on ranch products worldwide. It reduces milk production, damages milk quality, increases treatment costs, and even leads to the premature elimination of animals. In addition, failure to take effective measures in time will lead to widespread disease. The key to reducing the losses caused by mastitis lies in the early detection of the disease. The application of deep learning with powerful feature extraction capability in the medical field is receiving increasing attention. The main purpose of this study was to establish a deep learning network for buffalo quarter-level mastitis detection based on 3054 ultrasound images of udders from 271 buffaloes. Two data sets were generated with thresholds of somatic cell count (SCC) set as 2 × 105 cells/mL and 4 × 105 cells/mL, respectively. The udders with SCCs less than the threshold value were defined as healthy udders, and otherwise as mastitis-stricken udders. A total of 3054 udder ultrasound images were randomly divided into a training set (70%), a validation set (15%), and a test set (15%). We used the EfficientNet_b3 model with powerful learning capabilities in combination with the convolutional block attention module (CBAM) to train the mastitis detection model. To solve the problem of sample category imbalance, the PolyLoss module was used as the loss function. The training set and validation set were used to develop the mastitis detection model, and the test set was used to evaluate the network's performance. The results showed that, when the SCC threshold was 2 × 105 cells/mL, our established network exhibited an accuracy of 70.02%, a specificity of 77.93%, a sensitivity of 63.11%, and an area under the receiver operating characteristics curve (AUC) of 0.77 on the test set. The classification effect of the model was better when the SCC threshold was 4 × 105 cells/mL than when the SCC threshold was 2 × 105 cells/mL. Therefore, when SCC ≥ 4 × 105 cells/mL was defined as mastitis, our established deep neural network was determined as the most suitable model for farm on-site mastitis detection, and this network model exhibited an accuracy of 75.93%, a specificity of 80.23%, a sensitivity of 70.35%, and AUC 0.83 on the test set. This study established a 1/4 level mastitis detection model which provides a theoretical basis for mastitis detection in buffaloes mostly raised by small farmers lacking mastitis diagnostic conditions in developing countries.

Dietary elaidic acid boosts tumoral antigen presentation and cancer immunity via ACSL5.

Immunomodulatory effects of long-chain fatty acids (LCFAs) and their activating enzyme, acyl-coenzyme A (CoA) synthetase long-chain family (ACSL), in the tumor microenvironment remain largely unknown. Here, we find that ACSL5 functions as an immune-dependent tumor suppressor. ACSL5 expression sensitizes tumors to PD-1 blockade therapy in vivo and the cytotoxicity mediated by CD8+ T cells in vitro via regulation of major histocompatibility complex class I (MHC-I)-mediated antigen presentation. Through screening potential substrates for ACSL5, we further identify that elaidic acid (EA), a trans LCFA that has long been considered harmful to human health, phenocopies to enhance MHC-I expression. EA supplementation can suppress tumor growth and sensitize PD-1 blockade therapy. Clinically, ACSL5 expression is positively associated with improved survival in patients with lung cancer, and plasma EA level is also predictive for immunotherapy efficiency. Our findings provide a foundation for enhancing immunotherapy through either targeting ACSL5 or metabolic reprogramming of antigen presentation via dietary EA supplementation.

NADPH oxidase 2 inhibitor GSK2795039 prevents doxorubicin-induced cardiac atrophy by attenuating cardiac sympathetic nerve terminal abnormalities and myocyte autophagy.

Doxorubicin is widely used for the treatment of human cancer, but its clinical use is limited by a cumulative dose-dependent cardiotoxicity. However, the mechanism of doxorubicin-induced cardiac atrophy and failure remains to be fully understood. In this study, we tested whether the specific NADPH oxidase 2 (Nox2) inhibitor GSK2795039 attenuates cardiac sympathetic nerve terminal abnormalities and myocyte autophagy, leading to the amelioration of cardiac atrophy and dysfunction in chronic doxorubicin-induced cardiomyopathy. Mice were randomized to receive saline, doxorubicin (2.5 mg/kg, every other day, 6 times) or doxorubicin plus GSK2795039 (2.5 mg/kg, twice a day, 9 weeks). Left ventricular (LV) total wall thickness and LV ejection fraction were decreased in doxorubicin-treated mice compared with saline-treated mice and the decreases were prevented by the treatment of the specific Nox2 inhibitor GSK2795039. The ratio of total heart weight to tibia length and myocyte cross-sectional area were decreased in doxorubicin-treated mice, and the decreases were attenuated by the GSK2795039 treatment. In doxorubicin-treated mice, myocardial Nox2 and 4-hydroxynonenal levels were increased, myocardial expression of GAP43, tyrosine hydroxylase and norepinephrine transporter, markers of sympathetic nerve terminals, was decreased, and these changes were prevented by the GSK2795039 treatment. The ratio of LC3 II/I, a marker of autophagy, and Atg5, Atg12 and Atg12-Atg5 conjugate proteins were increased in doxorubicin-treated mice, and the increases were attenuated by the GSK2795039 treatment. These findings suggest that inhibition of Nox2 by GSK2795039 attenuates cardiac sympathetic nerve terminal abnormalities and myocyte autophagy, thereby ameliorating cardiac atrophy and dysfunction after chronic doxorubicin treatment.

AS602801 treatment suppresses breast cancer metastasis to the brain by interfering with gap-junction communication by regulating Cx43 expression.

AS602801 has been reported as a potential drug candidate against brain metastasis by suppressing the gap-junction communication between lung cancer stem cells and astrocytes. In this study, we aimed to study the molecular mechanism underlying the role of AS602801 in the treatment of brain metastasis in breast cancer. We utilized female athymic BALB/c nude mice and MDA-MB-231/BT-474BR cells to establish experimental models. Polymerase chain reaction assays were performed to observe changes in the connexin 43 (Cx43) messenger RNA (mRNA) and c-Jun N-terminal kinase (JNK) mRNA levels. Dye transfer assay was used to observe the effect of AS602801 on cell-cell communication. An organotypic blood-brain barrier (BBB) model was utilized to observe the effect of AS602801 on transmigration through the BBB barrier. MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay and flow cytometry were performed to evaluate the proliferation and apoptosis of breast cancer cells co-cultivated with astrocytes. AS602801 inhibited the upregulation of Cx43 and JNK in brain metastasized breast cancer cells in a dose-dependent manner. Also, AS602801 significantly decreased the dye transfer rate from astrocytes to breast cancer cells, indicating the inhibitory effect of AS602801 on cell-cell communication. The transmigration ability of breast cancer cells co-cultured with astrocytes was decreased by AS602801. Furthermore, AS602801 reduced the elevated Cx43/JNK mRNA expression in the co-astrocyte group while suppressing the increased proliferation and promoting the decreased apoptosis of breast cancer cells co-cultivated with astrocytes. AS602801 also suppressed the brain metastasis of breast cancer cells and increased mouse survival. AS602801 downregulates the expressions of JNK and Cx43 to suppress the gap-junction activity. AS602801 also inhibits the communication between breast cancer cells and astrocytes, thus contributing to the treatment of brain metastasis in breast cancer.

Cotton fiber-based dressings with wireless electrical stimulation and antibacterial activity for wound repair.

Although cotton dressing is one of the most commonly used wound management materials, it lacks antimicrobial and healing-promoting activity. This work developed a multilayer electroactive composite cotton dressing (Ag/Zn@Cotton/Paraffin) with exudate-activated electrical stimulation and antibacterial activity by the green and sustainable magnetron-sputtering and spraying methods. The inner hydrophilic layer of the cotton dressing was magnetron sputtered with silver/zinc galvanic couple arrays (Ag/Zn), which can be activated by wound exudate, generating an electrical stimulation (ES) into the wound. The Ag/Zn@Cotton showed efficient antibacterial activities against S. aureus and E. coli. Meanwhile, the paraffin-sprayed outer surface showed excellent antibacterial adhesion rates for S. aureus (99.82 %) and E. coli (97.92 %). The in vitro cell experiments showed that the ES generated by Ag/Zn@Cotton/Paraffin increased the migration of fibroblasts, and the in vivo mouse model indicated that the Ag/Zn@Cotton/Paraffin could enhance wound healing via re-epithelialization, inflammatory inhibition, collagen deposition, and angiogenesis. MTT method and live/dead staining showed that Ag/Zn@Cotton/Paraffin had no significant cytotoxic effects. This work may shed some light on designing and fabricating multi-functional electroactive composited dressings based on traditional biomedical textiles.

Identification of Milk Adulteration in Camel Milk Using FT-Mid-Infrared Spectroscopy and Machine Learning Models.

Camel milk, esteemed for its high nutritional value, has long been a subject of interest. However, the adulteration of camel milk with cow milk poses a significant threat to food quality and safety. Fourier-transform infrared spectroscopy (FT-MIR) has emerged as a rapid method for the detection and quantification of cow milk adulteration. Nevertheless, its effectiveness in conveniently detecting adulteration in camel milk remains to be determined. Camel milk samples were collected from Alxa League, Inner Mongolia, China, and were supplemented with varying concentrations of cow milk samples. Spectra were acquired using the FOSS FT6000 spectrometer, and a diverse set of machine learning models was employed to detect cow milk adulteration in camel milk. Our results demonstrate that the Linear Discriminant Analysis (LDA) model effectively distinguishes pure camel milk from adulterated samples, maintaining a 100% detection rate even at cow milk addition levels of 10 g/100 g. The neural network quantitative model for cow milk adulteration in camel milk exhibited a detection limit of 3.27 g/100 g and a quantification limit of 10.90 g/100 g. The quantitative model demonstrated excellent precision and accuracy within the range of 10-90 g/100 g of adulteration. This study highlights the potential of FT-MIR spectroscopy in conjunction with machine learning techniques for ensuring the authenticity and quality of camel milk, thus addressing concerns related to food integrity and consumer safety.

Sustainable supplier selection based on VIKOR with single-valued neutrosophic sets.

Considering economic, environmental, and social issues, the sustainability of the supply chain has drawn considerable attention due to societal and environmental changes within the supply chain network. The strategic study of the entire supply chain process and maximizing an organization's competitive advantage depend heavily on supplier selection based on sustainable indicators. Selecting sustainable suppliers for the supply chain is challenging since it is a multi-criteria decision-making (MCDM) problem with significant uncertainty in the decision-making process. This study uses the VlseKriterijumska Optimizacija I Kompromisno Resenje (VIKOR) technique and single-valued neutrosophic sets (SVNS) to deal with the challenge of choosing a sustainable supplier with insufficient information. This method reduces the influence of personal experience and preference on the final evaluation results and the problem of excessive individual regret caused by factor correlation and improves the consistency of evaluation results. Finally, the method's success and adaptability are demonstrated by sensitivity analysis and additional comparison analysis, and the benefits and drawbacks of the suggested framework are examined. Compared to other approaches, it can assist decision-makers in communicating fuzzy and uncertain information, offering a perspective and approach for MCDM in the face of such situations, and helping them select suppliers of high caliber and who practice sustainable business practices.

New insights into transcriptome variation during cattle adipocyte adipogenesis by direct RNA sequencing.

We performed direct RNA sequencing (DRS) together with PCR-amplified cDNA long and short read sequencing for cattle adipocyte at different stages. We proved that the DRS was with advantages to avoid artificial transcripts and questionable exitrons. Totally, we obtained 68,124 transcripts with information of alternative splicing, poly (A) length and mRNA modification. The number of transcripts for adipogenesis was expanded by alternative splicing, which lead regulation mechanisms far more complex than ever known. We detected 891 differentially expressed genes (DEGs). However, 62.78% transcripts of DEGs were not significantly differentially expressed, and 248 transcripts showed opposite changing directions with their genes. The poly (A) tail became globally shorter in differentiated adipocyte than in primary adipocyte, and had a weak negative correlation with gene/transcript expression. Moreover, the study of different mRNA modifications implied their potential roles in gene expression and alternative splicing. Overall, our study promoted better understanding of adipogenesis mechanisms in cattle adipocytes.

Relationships between body- and udder-related type traits with somatic cell counts and potential use for an early selection method for water buffaloes (Bubalus bubalis).

Water buffalo milk is a reliable source of high-quality nutrients; however, the susceptibility of mastitis in buffaloes must be taken into consideration. An animal with somatic cell count (SCC) of greater than 250,000 cells/mL is reported to be likely to have mastitis which has serious adverse effects on animal health, reproduction, milk yield, and milk quality. Type traits (TTs) of water buffalo can affect SCC in animal milk to some extent, but few reports on the correlation between SCC and TTs are available. In this study, a total of 1908 records collected from 678 water buffaloes were investigated. The general linear model was used to identify factors associated with phenotypic variation of the somatic cell score (SCS) trait, including parity, lactation length, calving year, and calving season as fixed effects. Using PROC CORR analysis method, taking calving year and lactation length as covariates, the correlation co-efficient between TT and SCS was obtained. Our results showed that correlation co-efficients between the 45 TTs with SCS ranged from 0.003 to 0.443 (degree of correlation). The correlation between udder traits and SCS was greater than that between body structure traits and SCS. Among udder traits, distance between teats (including front and rear teat distance [r = 0.308], front teat distance [r = 0.211], and teat crossing distance [r = 0.412]) and teat circumference (r = 0.443) had the highest correlation with SCS, followed by the leg traits including rear leg height (r = -0.354) and hock bend angle (r = -0.170). Animal with high rear legs (>48 cm) and short teat crossing distance (<17 cm), and narrow teat circumference (<11 cm) exhibited low SCS. Using four nonlinear models (Von Bertalanffy, Brody, Logistic, and Gompertz), the optimal growth curves of the TTs highly correlated with the SCS (rear leg height and teat crossing distance) were fitted, and the correction co-efficients of these two TTs rear leg height and teat crossing distance of animal from young age (2 mo old) to first lactation (35 mo old) were attained for establishment of early selection method for water buffaloes with low SCS. This study provides theoretical support for early selection of low-SCS water buffaloes and lays a foundation for improving milk quality and promoting healthy development of water buffalo's dairy industry.

Some type traits (TTs) have been reported to affect somatic cell count (SCC) in bovine milk, which result in mastitis to some extent, but the correlation between SCC and TTs of water buffalo has been poorly understood. Here, a total of 1908 records from 678 buffaloes were investigated. The correlation between 45 TTs and somatic cell score (SCS) was analysed, and the optimal growth curves of TTs highly correlated with SCS were fitted. Our result showed that high rear legs (>48 cm) and short teat crossing distance (<17 cm), and narrow teat circumference (<11 cm) were correlated with low SCS. We obtained correction co-efficients for two TTs highly correlated with SCS of water buffalos from young age (2 mo old) to first lactation (35 mo) by fitting the optimal growth curve for rear leg height and teat crossing distance. This study provides theoretical support for early selection for water buffaloes that are less susceptible to mastitis, and lays a foundation for improving milk quality and promoting healthy development of water buffalo dairy industry.

The FSH-mTOR-CNP signaling axis initiates follicular antrum formation by regulating tight junction, ion pumps, and aquaporins.

The initial formation of the follicular antrum (iFFA) serves as a dividing line between gonadotropin-independent and gonadotropin-dependent folliculogenesis, enabling the follicle to sensitively respond to gonadotropins for its further development. However, the mechanism underlying iFFA remains elusive. Herein, we reported that iFFA is characterized by enhanced fluid absorption, energy consumption, secretion, and proliferation and shares a regulatory mechanism with blastula cavity formation. By use of bioinformatics analysis, follicular culture, RNA interference, and other techniques, we further demonstrated that the tight junction, ion pumps, and aquaporins are essential for follicular fluid accumulation during iFFA, as a deficiency of any one of these negatively impacts fluid accumulation and antrum formation. The intraovarian mammalian target of rapamycin-C-type natriuretic peptide pathway, activated by follicle-stimulating hormone, initiated iFFA by activating tight junction, ion pumps, and aquaporins. Building on this, we promoted iFFA by transiently activating mammalian target of rapamycin in cultured follicles and significantly increased oocyte yield. These findings represent a significant advancement in iFFA research, further enhancing our understanding of folliculogenesis in mammals.

Electroactive nanofibrous membrane with temperature monitoring for wound healing.

Developing functional dressings for promoting cellular activities and monitoring the healing progress is receiving increasingly widespread attention. In this study, Ag/Zn electrodes were deposited on the surface of a polylactic acid (PLA) nanofibrous membrane which can mimic the extracellular matrix. When wetted by wound exudate, the Ag/Zn electrodes could generate an electric stimulation (ES), promoting the migration of fibroblasts that heal wounds. Moreover, the Ag/Zn@PLA dressing showed excellent antibacterial activity against E. coli (95%) and S. aureus (97%). The study found that the electrostatic (ES) effect and the release of metal ions mainly contribute to the wound healing properties of Ag/Zn@PLA. In vivo mouse models demonstrated that Ag/Zn@PLA could promote wound healing by improving re-epithelialization, collagen deposition, and neovascularization. Additionally, the integrated sensor within the Ag/Zn@PLA dressing can monitor the wound site's temperature in real-time, providing timely information on wound inflammatory reactions. Overall, this work suggests that combining electroactive therapy and wound temperature monitoring may provide a new strategy for designing functional wound dressings.

Infrared Thermography as a Potential Non-Invasive Tool for Estrus Detection in Cattle and Buffaloes.

The productivity of dairy animals has significantly increased over the past few decades due to intense genetic selection. However, the enhanced yield performance of milk animals caused a proportional increase in stress and compromised reproductive efficiency. Optimal reproductive performance is mandatory for the sustainable production of dairy animals. Reproductive efficiency is marked by proper estrus detection and precise breeding to achieve maximum pregnancies. The existing conventional methods of estrus detection are somewhat labor intensive and less efficient. Similarly, the modern automated methods that rely on detecting physical activity are expensive, and their efficiency is affected by factors such as type of housing (tie stall), flooring, and environment. Infrared thermography has recently emerged as a technique that does not depend on monitoring physical activity. Furthermore, infrared thermography is a non-invasive, user-friendly, and stress-free option that aids in the detection of estrus in dairy animals. Infrared thermography has the potential to be considered a useful non-invasive tool for detecting temperature fluctuations to generate estrus alerts without physical contact in cattle and buffaloes. This manuscript highlights the potential use of infrared thermography to understand reproductive physiology and practical implementation of this technique through discussing its advantages, limitations, and possible precautions.

FHL2 deficiency impairs follicular development and fertility by attenuating EGF/EGFR/YAP signaling in ovarian granulosa cells.

Female subfertility is an increasing reproductive issue worldwide, which is partially related to abnormal ovarian follicular development. Granulosa cells (GCs), by providing the necessary physical support and microenvironment for follicular development, play critical roles in maintaining female fertility. We previously showed that ectopic expression of four and a half LIM domains 2 (FHL2) promoted ovarian granulosa cell tumor progression. However, its function in follicular development and fertility remains unknown. Here, we confirmed that FHL2 is highly expressed in human and mouse ovaries. FHL2 immunosignals were predominantly expressed in ovarian GCs. A Fhl2 knockout (KO) mouse model was generated to examine its roles in follicular development and fertility. Compared with wildtype, knockout of Fhl2 significantly decreased female litter size and offspring number. Furthermore, Fhl2 deficiency reduced ovarian size and impaired follicular development. RNA-sequencing analysis of GCs isolated from either KO or WT mice revealed that, Fhl2 deletion impaired multiple biological functions and signaling pathways, such as Ovarian Putative Early Atresia Granulosa Cell, ErbB, Hippo/YAP, etc. In vitro studies confirmed that FHL2 silencing suppressed GCs growth and EGF-induced GCs proliferation, while its overexpression promoted GC proliferation and decreased apoptosis. Mechanistic studies indicated that FHL2, via forming complexes with transcriptional factors AP-1 or NF-κB, regulated Egf and Egfr expression, respectively. Besides, FHL2 depletion decreased YAP1 expression, especially the active form of YAP1 (nuclear YAP1) in GCs of growing follicles. EGF, serving as an autocrine/paracrine factor, not only induced FHL2 expression and nuclear accumulation, but also stimulated YAP1 expression and activation. Collectively, our study suggests that FHL2 interacts with EGFR and Hippo/YAP signaling to regulate follicular development and maintain fertility. This study illuminates a novel mechanism for follicular development and a potential therapeutic target to address subfertility.

Benefits of using Double-Ovsynch versus presynch-ovsynch are affected by environmental heat in primiparous holstein lactating cows.

Optimized reproduction management enhances fertility of dairy cows, and thus improves their milk production efficiency. Comparing different synchronization protocols under variable ambient conditions would be conducive to protocol selection and production efficiency improvement. Here, 9538 primiparous Holstein lactating cows were enrolled to either Double-Ovsynch (DO) or Presynch-Ovsynch (PO) to determine the outcomes under different ambiences. We found that averaged THI of 21-days before the first service (THI-b) was the best indicators in a total of 12 environmental indexes to explain changes in conception rate. And the conception rate decreased linearly in DO treated cows when THI-b was over 73, whereas the threshold was 64 in cows subjected to PO. Compared with PO treated cows, DO increased conception rate by 6%, 13% and 19%, when THI-b was lower than 64, from 64 to 73, and over 73, respectively. Furthermore, employing treatment of PO would lead greater risk for cows staying open compared with DO when THI-b below 64 (hazard ratio, 1.3) and over 73 (hazard ratio, 1.4). Most importantly, calving intervals were 15 days shorter in DO treated cows compared PO when THI-b over 73, while no difference was detected when THI-b below 64. In conclusion, our results supported that, fertility of primiparous Holstein cows could be improved by employing DO, especially in hot weather (THI-b ≥ 73), and the benefits of DO protocol were abated under cool conditions (THI-b < 64). Considering the impacts of environmental heat load is necessary to determine reproductive protocols for commercial dairy farm.

Lactational exposure to Deoxynivalenol causes mammary gland injury via inducing inflammatory response and impairing blood-milk barrier integrity in mice.

Lactation is a unique physiological process to produce and secrete milk. Deoxynivalenol (DON) exposure during lactation has been demonstrated to affect adversely the growth development of offspring. However, the effects and potential mechanism of DON on maternal mammary glands remain largely unknown. In this study, we found the length and area of mammary glands were significantly reduced after DON exposure on lactation day (LD) 7 and LD 21. RNA-seq analysis results showed that the differentially expressed genes (DEGs) were significantly enriched in acute inflammatory response and HIF-1 signaling pathway, which led to an increase of myeloperoxidase activity and inflammatory cytokines. Furthermore, lactational DON exposure increased blood-milk barrier permeability by reducing the expression of ZO-1 and Occludin, promoted cell apoptosis by upregulating the expression of Bax and cleaved Caspase-3 and downregulating the expression of Bcl-2 and PCNA. Additionally, lactational DON exposure significantly decreased serum concentration of prolactin, estrogen, and progesterone. All these alterations eventually resulted in a decrease of ß-casein expression on LD 7 and LD 21. In summary, our findings indicated that lactational exposure to DON caused lactation-related hormone disorder and mammary gland injury induced by inflammatory response and blood-milk barrier integrity impairment, ultimately resulting in lower production of ß-casein.

Establishment and Validation of Fourier Transform Infrared Spectroscopy (FT-MIR) Methodology for the Detection of Linoleic Acid in Buffalo Milk.

Buffalo milk is a dairy product that is considered to have a higher nutritional value compared to cow's milk. Linoleic acid (LA) is an essential fatty acid that is important for human health. This study aimed to investigate and validate the use of Fourier transform mid-infrared spectroscopy (FT-MIR) for the quantification of the linoleic acid in buffalo milk. Three machine learning models were used to predict linoleic acid content, and random forest was employed to select the most important subset of spectra for improved model performance. The validity of the FT-MIR methods was evaluated in accordance with ICH Q2 (R1) guidelines using the accuracy profile method, and the precision, the accuracy, and the limit of quantification were determined. The results showed that Fourier transform infrared spectroscopy is a suitable technique for the analysis of linoleic acid, with a lower limit of quantification of 0.15 mg/mL milk. Our results showed that FT-MIR spectroscopy is a viable method for LA concentration analysis.