Animal Trials Research Articles

Effects of Dietary Supplementation with Lactobacillus reuteri Postbiotics on Growth Performance, Intestinal Flora Structure and Plasma Metabolome of Weaned Piglets

Probiotics and their postbiotics have the potential to improve the health and growth performance of piglets, which has brought them widespread attention in the post-antibiotic era. In the present study, the effects of dietary supplementation of Lactobacillus reuteri postbiotics on the growth performance, intestinal flora structure and plasma metabolome of weaned piglets were investigated. A total of 816 healthy male piglets with uniform weight were divided into two treatment groups: piglets in the control (CTR) group were fed with a basic diet, and the ones in the LAC group were fed with the basic diet supplemented with 500 mg/kg Lactobacillus reuteri postbiotics. There were six replicates in each group and 68 piglets in each replicate. The animal trial lasted for 30 days. The feces and blood of piglets were collected for investigation, and the growth performance during the trial was counted. Our outcomes show that dietary supplementation with Lactobacillus reuteri postbiotics had no effect on the growth performance of piglets; however, it reduced the mortality rate of piglets by 6.37%. The levels of total superoxide dismutase in the serum, propionic acid and butyric acid in the feces were elevated, and the content of malondialdehyde in the serum was decreased with Lactobacillus reuteri postbiotics-treated piglets (p < 0.05). The fecal flora sequencing results show that the relative abundance of Firmicutes and monoglobus was upregulated, and the relative abundance of Bacteroides was downregulated with Lactobacillus reuteri postbiotics-treated piglets (p < 0.05). In addition, the levels of propionic acid and butyric acid in the feces were positively correlated with the relative abundance of Firmicutes and negatively correlated with the relative abundance of Bacteroides (p < 0.05). The plasma metabolome results show that dietary supplementation with Lactobacillus reuteri postbiotics raised the level of coenzyme Q10 in the serum, and the abundance of coenzyme Q10 was positively correlated with the relative abundance of Firmicutes and the level of total superoxide dismutase in the serum. In conclusion, dietary supplementation with Lactobacillus reuteri postbiotics contributed to improving the antioxidant function and reducing the mortality of piglets by regulating the structure of intestinal flora and upregulating the content of coenzyme Q10 in serum.

Associations between lameness and the metabolic and hormonal profiles in dairy cows

Dairy cows lameness represents a significant welfare issue for the dairy industry. The aim of this study was to investigate metabolic and hormonal parameters associated with lameness in cows. The study included two treatment groups of 10 multiparous Holstein–Friesian dairy cows each. The control group consisted of cows with locomotion score 1 and in the experimental group cows with locomotion scores 3 and 4 were included. Milk, urine and blood samples were collected to determine the fat, protein, urea, creatinine, electrolytes and copeptin vasopressin (AVP) content. The milk fat, protein and energy-corrected milk (ECM) levels were higher in lame animals compared with non-lame cows. The blood urea nitrogen (BUN) and BUN/serum creatinine ratio were lower at the start of the trial in lame animals suggesting a metabolic acidosis state. There were significantly lower serum Na levels in the lame cows, from the start until the end of the trial (p < 0.011), as well as between lame and non-lame groups at the end of the trial (p = 0.041). A lower copeptin AVP was detected in lame compared with non-lame cows at the end of the trial (p = 0.004). The decrease of serum Na levels as the comfort improves indicates that the animals started to recover from the effect of cortisol on mineralocorticoid receptors. Our data also indicates that the BUN and BUN/serum creatinine ratios are associated with lameness. Further investigations involving a larger number of cows would be recommended to confirm these results and to evaluate these parameters in the early stages of lameness.

Phytochemical Profiling and Biological Activities of Nigella sativa Vinegar Extract: An Insight Into Hypoglycemic Potential Using In Vivo and In Silico Studies.

The current study was conducted to characterize the vinegar extract of Nigella sativa and evaluate its biological activities using in vitro and in vivo studies. The N. sativa extract (NSE) was prepared by macerating seeds in a mixture of water and synthetic vinegar(1:10). The antioxidant potential of NSE was assessed, revealing high total phenolic contents (431.66mg/100g), total flavonoid contents (73.45mg/100g), total antioxidant capacity (118.55mg/100g), and strong radical scavenging potential (89.67%). The anticancer activity of NSE showed insignificant cytotoxic effects on HepG2 liver cell lines (96.61±3.00). An in vivo animal trial was performed, and NSE significantly (p<0.05) lowered the blood glucose (-47.71%), total cholesterol (-37.86%), total glycerides (39.42%), low-density lipoproteins (-29.36%), and very low-density lipoproteins (-30.60%), while increasing the high-density lipoproteins (26.92%) in alloxan-induced diabetic rats. GC-MS analysis of NSE revealed a diverse range of bioactive compounds, including alkaloids, phenolics, and flavonoids. In silico studies using molecular docking simulations showed that pyrrolo[1,2-a] pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl) demonstrated a strong inhibitory action (6.6kcal/mol) against alpha-amylase enzyme. These findings suggest that NSE is a promising natural remedy for managing diabetes and related metabolic disorders, and its potential as a nutraceutical ingredient in functional food applications is significant.

Applications of Semaglutide for Various Diseases

Semaglutide is a gluten-like peptide 1 (GLP-1) agonist which is responsible for improving blood sugar control and losing weight. It is clinically approved by the FDA for treatment of type 2 diabetes mellitus and is designed in subcutaneous or oral dosage form. Some other diseases share common conditions with obesity and diabetes such as a high blood sugar level and insulin resistance. These factors along with some neuron issues are responsible for neurodegenerative diseases. Obesity and diabetes are two common worldwide health issues. A number of trials have been conducted to estimate the prevalence and drug effects. Scientists also study related diseases such as Non-alcoholic fatty liver disease to verify the usefulness of treatments. Semaglutide is one of the drugs being measured whether has a potential effect on these diseases. Some pre-clinical studies have indicated its therapeutic potential in non-alcoholic steatohepatitis and neurodegenerative disorders. Semaglutide has proved to be effective in treating lots of disorders, especially diabetes and obesity. However, for other diseases, it still remains uncertain even if there are some studies already tested the effectiveness. Much more detailed and considerate studies should be conducted to verify the impact. Semaglutide, the GLP-1 receptor agonist, has shown a positive impact in treating different diseases. Researchers have tested hepatoprotective and neuroprotective activity in animal and human trials to get more reliable data. It turns out that semaglutide improves insulin resistance and reduces blood sugar levels. It also protects dopaminergic neurons and reduces aggregated α-synuclein which improves Parkinson’s disease and Alzheimer’s disease conditions.

Effects of Lactiplantibacillus plantarum and Galactooligosaccharide Administered In Ovo on Hatchability, Chick Quality, Performance, Caecal Histomorphology and Meat Quality Traits of Broiler Chickens.

The presented study explored the promising alternatives of in ovo injection with Lactiplantibacillus plantarum (LP) and galactooligosaccharide (GOS) in the poultry industry. The study aimed to assess the effects of probiotic and prebiotic on various aspects of poultry production. The study involved 300 Ross broiler eggs, individually candled on Day 7 of embryonic development. The eggs were sorted into four groups: negative control (no injection), positive control (0.9% physiological saline injection), GOS 3.5 mg/egg and LP 1 × 106 CFU/egg. The groups used during the incubation period were the same for the animal trial; each pen/group had 25 chickens. At the end of the experiment, 8 chickens from each group were slaughtered for tissue sample collection and 12 chickens were slaughtered to determine slaughter yield, carcass and meat quality. All data were analysed by one-way ANOVA or repeated measured ANOVA except for the parameters that did not meet the assumption of normality, the Kruskal-Wallis test (Dunn's test) was used. Key findings revealed that hatchability remained unaffected across groups, indicating the safety of the in ovo injections. Both LP and GOS enhanced chick quality, as evidenced by improved body weight, Pasgar score and chick length. The in ovo administration of LP increased the body weight of the chickens during the first-week post-hatch (7 days of age) without impacting feed intake and feed conversion ratio in the later stages. The study demonstrated no adverse effects on meat quality due to the in ovo injection of LP and GOS. Additionally, a positive impact on caecal histomorphology was observed and early gut colonization of beneficial bacteria (Lactobacillus spp. and Bifidobacteria spp.) indicated potential benefits for intestinal health in broilers. In conclusion, the in ovo inoculation of 1 × 106 LP and 3.5 mg of GOS per egg increased the relative abundance of Lactobacillus spp. and Bifidobacterium spp. and showcased promising enhancements in chick quality without compromising growth performance, meat quality and caecal histomorphology. These findings suggest a positive outlook for these substances as a viable alternative for improving poultry health and productivity.

Rutin Synergizes with Colistin to Eradicate Salmonellosis in Mice by Enhancing the Efficacy and Reducing the Toxicity.

The wide dissemination of multidrug-resistant (MDR) Gram-negative bacteria poses a significant global health and security concern. As developing new antibiotics is generally costly, fastidious, and time-consuming, there is an urgent need for alternative therapeutic strategies to address the gap in antibiotic discovery void. This study aimed to investigate the activity of colistin (CS) in combination with a natural product, rutin (RT), to combat against Salmonella Typhimurium (S. Tm) in vitro and in vivo. The results showed that a combination with RT enabled the potentiation of CS efficacy. Further mechanistic analysis indicated that RT disrupted iron homeostasis to inactivate the PmrA/PmrB system, thereafter reducing the bacterial membrane modifications for enhancing CS binding. Besides enhancing bactericidal activity of CS, RT was also observed to mitigate the CS-induced nephrotoxicity, by which the dosing limitation of CS was overcome for better pathogen clearance. The animal trial eventually confirmed the in vivo synergistic interaction of RT with CS to treat the bacterial infection. To sum up, the present study uncovered the potential of RT as a viable adjuvant of CS to eradicate the infection and protect the hosts, which might serve as a promising alternative to combat infections caused by MDR Gram-negative bacteria.

Design and Experiment of a PLC-Based Intelligent Thermal Insulation Box for Nursing Piglets.

Local heating of the activity area for nursing piglets is crucial for piglet health and the energy efficiency of barn climate control. Traditional heating methods using lamps or covers lack precise control, result in significant energy waste, and cannot be dynamically adjusted according to piglet age or changing environmental temperatures. To address these issues, this study designed a Programmable Logic Controller (PLC)-based thermal insulation box for nursing piglets, utilizing a strip heater instead of the conventional round heating lamp. The design incorporates a movable thermal insulation box that dynamically adjusts the heater's power based on the real-time monitoring of environmental temperatures and target temperatures specific to piglet age. First, in a controlled laboratory environment, the study tested and compared the spatial temperature uniformity, temporal stability, and power consumption of the new thermal insulation box versus traditional heating methods. Subsequently, animal trials were conducted in a farrowing barn using eight sows with similar farrowing dates as test subjects. The new thermal insulation box was installed in one group, and the traditional heating lamp in the control group. During the trial, ambient temperature, insulation area temperature, piglet behavior, growth performance, and power consumption were recorded. The results showed that compared to the control group, the new system reduced average temperature fluctuations in the insulation area by 31.6% and spatial temperature variation by 78.3%. During animal trials, the average temperatures directly under the heater for the new system versus the control in the insulation area were 39.7 ± 0.2 °C and 30.2 ± 1.4 °C in the first week, 40.9 ± 0.5 °C and 31.6 ± 0.7 °C in the second week, and 32.3 ± 1.5 °C and 28.6 ± 1.7 °C in the third week-significantly (p < 0.05) higher in the test group. The new system also reduced total energy consumption by 58.3%. The usage rate of the thermal insulation area by piglets in the test and control groups was 47.5 ± 5.3% and 42.1 ± 6.6%. The daily weight gain of piglets in the test group was 9.8% higher than that of the control group, also significantly (p < 0.05) higher. This intelligent thermal insulation box enables precise and dynamic temperature control, reducing heating energy consumption and supporting improved piglet health and welfare.

The Impact of Hyperbaric Oxygen Therapy on Functional and Structural Plasticity in Rats With Spinal Cord Injury.

Spinal cord injury (SCI) can result in sensory and locomotor function loss below the injured segment. Hyperbaric oxygen therapy (HBOT) has been proven to alleviate SCI. This study aims to establish a reproducible rat model of SCI and investigate the impact of HBOT on alterations in brain neuronal activity and neuromotor function in this experimental rat SCI model using resting-state functional magnetic resonance imaging (rs-fMRI). This is a prospective randomized controlled animal trial. A total number of 27 female SD rats were randomly divided into three groups: sham (n=9), SCI (n=9), and HBO (n=9). rs-fMRI was utilized to assess regional homogeneity (ReHo) values and functional connectivity (FC) strength over the whole brain with the motor cortex as seeds. Correlation between neuroimaging characteristics and behavioral assessment was calculated. We examined Nissl body, NeuN, and caspase-3 expression in relevant brain regions. Following SCI, reduced ReHo values were observed in the left primary somatosensory cortex, left striatum, right agranular insular cortex, and partial cortex in the limbic system, which was reversed after HBOT. HBOT could increase FC strength between the motor cortex and other brain regions, including the left secondary motor cortex, right basal forebrain region, bilateral primary somatosensory cortex, bilateral thalamus, and another partial cortex in the limbic system. BBB scale scores showed that HBOT promoted motor function recovery in SCI rats. The ReHo and FC values in all positive clusters were positively correlated with BBB scores. By histopathological analysis, our study found that HBOT could reduce apoptotic proteins, increase the number of neurons, and protect neuronal function in brain regions with significant ReHo and FC alteration in SCI rats. This study reveals that HBOT facilitates functional and structural plasticity in the brain, contributing to the recovery of motor function in rats with SCI.

Development and Clinical Application of High-Magnification Electronic Endoscope

This article introduces a high-magnification electronic endoscope system that utilizes a continuous zoom optical design, enabling high-magnification imaging at the cellular level. By selecting an appropriate initial retrofocus structure, the optimized modulation transfer function (MTF) curve of the zoom objective lens approaches the diffraction limit. Additionally, the tip and operation parts of the system incorporate modular gas-tight sealing and multiple layers of protective structure. The design employs efficient sealing materials and precise packaging techniques to ensure the system's sealing performance. The experimental results indicate that, under conventional imaging, the resolution is equivalent to 39.37 μm in object space, and the field of view angle is larger than 140 degrees. At magnified imaging, the resolution is equivalent to 2.78 μm in object space, and the observed actual field of view is greater than 800 μm×700 μm. When used with a 26-inch display monitor, the high-magnification electronic endoscope can achieve 500× optical magnification, allowing for the observation of features at the cellular level. Currently, this product has successfully achieved mass production and has undergone multiple animal trials, demonstrating its wide range of potential applications and significant clinical value.

The Potential Effect of Daun Sirih (Piperaceae Leaf) on the Burn Wound Healing: A Literature Review

Introduction: Burn injuries, particularly among children and the elderly, result from heat, chemicals, or electricity, leading to skin and tissue damage. The wound healing process involves several stages, and infections from bacteria such as Pseudomonas aeruginosa and Staphylococcus aureus can complicate healing. While burns are typically treated with antiseptics, traditional remedies like Piper betle (Piperaceae) are commonly used in Indonesia. Compounds in Piper betle—including flavonoids, saponins, and tannins—possess antioxidant, antibacterial, and anti-inflammatory properties that may enhance burn wound healing. Methods: This literature review synthesized data from various open-access online databases to evaluate the effects of Piper betle on burn wound healing. Results and Discussion: Piper betle, a medicinal plant from the Piperaceae family, accelerates wound healing through several mechanisms. Flavonoids form complexes with proteins, damaging bacterial cells; saponins enhance collagen production, and tannins disrupt bacterial cell walls. Studies confirm that Piper betle reduces inflammation and fosters tissue regeneration via antioxidant activity. Animal trials demonstrate significant improvements in burn wound healing, including faster wound contraction and recovery. Conclusion: Piper betle shows promise in accelerating burn wound healing due to its bioactive compounds. While existing studies support its effectiveness, further research is needed to assess the long-term safety and potential toxicity of Piperaceae-based treatments.

Efficacy of Invasive and Non-Invasive Methods in Orthodontic Tooth Movement Acceleration: A Systematic Review

Objective: The aim of this review was to evaluate the currently available scientific evidence on the efficacy of different methods as accelerators of tooth movement during orthodontic treatment: corticotomies, piezocision, micro-osteoperforations (MOP), photobiomodulation (LLLT and LED laser) and microvibrations. Search Methods: A comprehensive search was performed in the PubMed, Google Scholar, Scopus and Medline databases in May 2024. Selection Criteria: We selected randomized controlled trials based on acceleration of tooth movement during orthodontic treatment. Articles that were not randomized controlled trials (RCTs), were not published in the last ten years or corresponded to animal trials as well as those dealing with orthognathic surgery, distraction osteogenesis, electric currents, pulsed electric fields and pharmacological approaches were excluded. Results: Twenty-three studies were included in this review. All trials show accelerated tooth movement after low-level laser application, and seven studies support the efficacy of surgically assisted orthodontic treatment with corticotomies, piezocision or MOP. No article indicates statistically significant differences between the application of microvibration during orthodontic treatment and conventional treatment. No negative effects on the periodontium, loss of dental vitality or serious root resorption were reported in any publication, except in a study carried out with MOP (with an increase in root resorption). Conclusions: There is some evidence that low-level laser therapy and surgical methods are effective techniques in accelerating tooth movement during orthodontic treatment, while the evidence is very weak for vibration.

Newcastle Disease Virus Expressing Cap Gene of Porcine Circovirus Type 2 Confers Protection in Mice and Induced Long-Lasting Neutralizing Antibodies in Pigs.

Background/Objectives: Porcine Circovirus 2 (PCV2) infection poses significant health and economic challenges to the global swine industry. The disease in pigs leads to lymphoid depletion, resulting in immunosuppression and increased susceptibility to co-infections with other bacterial and viral pathogens. This study evaluated the efficacy of two novel recombinant Newcastle disease virus (NDV) strain R2B vectored vaccines that express the cap gene of PCV2 alone and along with the transmembrane and cytoplasmic tail (TMCT) domains of the NDV F gene. The efficacy of the vaccine candidates was studied in mouse and pig models. Methods: Six-week-old BALB/c mice were divided into five groups and immunized intramuscularly three times at 14-day intervals with various vaccine candidates, namely rNDV-R2B-PCVcap-TMCT, rNDV-R2B-PCVcap, and CircoFLEX commercial vaccine, along with controls. Following immunization and PCV2d virus challenge, multiple assays assessed the immune responses in animal trials. In the pig animal trial, pigs were divided into four groups: a control group (PBS), NDV-vectored PCVcap-TMCT group, NDV-vectored-PCVcap group, and CircoFLEX vaccine group. Pigs were immunized intramuscularly twice at 28-day intervals. Blood samples were collected at regular intervals over 70 days to evaluate the humoral and cell-mediated immune responses. Results: Both mice and pigs' trials indicated that the NDV-vectored PCV2 cap-TMCT vaccine candidate elicited superior immune responses. In mice, the rNDV-R2B-PCVcap-TMCT group showed enhanced humoral and cellular immunity, increased PCV2-specific antibody levels, higher CD4+/CD8+ ratio, elevated IFN-γ and TNF-α levels, decreased IL-10 levels, reduced viral loads, and minimal histopathological changes. In pigs, the NDV-vectored PCVcap-TMCT group demonstrated better antibody responses, cytokine profiles (IFN-γ and IL-10), and higher levels of PCV2-specific neutralizing antibodies against the PCV2a, PCV2b and PCV2d genotypes when compared to other groups. Conclusions: These findings suggest NDV-vectored PCVcap-TMCT vaccine candidate, expressing the cap gene of PCV2 along with the TMCT domain, offers a promising alternative for protecting against PCV2 infection, potentially addressing the challenges posed by emerging PCV2 strains in the swine industry.

Research on the TSPAN6 regulating the secretion of ADSCs-Exos through syntenin-1 and promoting wound healing

BackgroundExosomes (Exos) from adipose-derived stem cells (ADSCs) have a high inclusion content and low immunogenicity, which helps to control inflammation and accelerate the healing of wounds. Unfortunately, the yield of exosomes is poor, which raises the expense and lengthens the treatment period in addition to impairing exosomes’ therapeutic impact. Thus, one of the key problems that needs to be resolved in the current exosome study is increasing the exosome yield.MethodsTetraspanin-6 (TSPAN6) overexpression and knockdown models of ADSCs were constructed to determine the number of exosomes secreted by each group of cells as well as the number of multivesicular bodies (MVBs) and intraluminal vesicles (ILVs) within the cells. Subsequently, the binding region of the interaction between TSPAN6 and syntenin-1 was identified using the yeast two-hybrid assay, and the interaction itself was identified by immunoprecipitation. Finally, cellular and animal studies were conducted to investigate the role of each class of exosomes.ResultsWhen compared to the control group, the number of intracellular MVBs and ILVs was significantly larger, and the number of ADSCsTSPAN6+-Exos was more than three times higher. However, TSPAN6’s ability to stimulate exosome secretion was reduced as a result of syntenin-1 knockdown. Additional yeast two-hybrid assay demonstrated that the critical structures for their interaction were the N-terminal, Postsynaptic density protein 95/Discs large protein/Zonula occludens 1 (PDZ1), and PDZ2 domains of syntenin-1, and the C-terminal of TSPAN6. In animal trials, the wound healing rate was best in the ADSCsTSPAN6+-Exos group, while cellular experiments demonstrated that ADSCsTSPAN6+-Exos better enhanced the proliferation and migration of human skin fibroblasts (HSFs) and human umbilical vein endothelial cells (HUVECs).ConclusionTSPAN6 stimulates exosome secretion and formation, as well as the creation of MVBs and ILVs in ADSCs. Syntenin-1 is essential for TSPAN6’s stimulation of ADSCs-Exos secretion. Furthermore, ADSCsTSPAN6+-Exos has a greater ability to support wound healing, angiogenesis, and the proliferation and migration of a variety of cells.

TMET-24. MULTI-OMIC CHARACTERIZATION OF THE TEMPORAL METABOLIC ADAPTATIONS UNDERPINNING THERAPY RESISTANCE AND TUMOR RECURRENCE IN GLIOBLASTOMA

Abstract Therapy resistance and tumor recurrence are barriers to achieving long-term survival in cancer patients, particularly evident in glioblastoma (GBM), known for its dismal 5-year survival rate below 10%. Despite aggressive treatment regimens, including maximal resection, alkylating chemotherapy, and radiation, the majority of GBM patients experience relapse within 7-9 months post-initial intervention. Recurrent GBM poses surgical challenges and resists subsequent treatments, leading to an almost universally fatal outcome and an average survival of 12-18 months. Metabolic networks can serve as ancient stress-protection mechanisms that operate independently of genomic changes, offering an initial line of defense against exogenous pressures. Although the role of metabolism in cancer has gained prominence, our understanding of the metabolic adaptations following chemoradiotherapy and their contributions to therapy resistance and tumor recurrence in highly aggressive GBM tumors remains incomplete. We postulate that metabolic adaptations play a pivotal role in facilitating therapy resistance and GBM recurrence. Exploring the metabolic dependencies of these persistent cells may unveil vulnerabilities for therapeutic exploitation to counteract chemoradiotherapy resistance. We established clinically relevant patient-derived in vitro and in vivo models mirroring the standard GBM treatment protocol. Using state-of-the-art single-cell and bulk multi-omics technologies, we profiled the temporal transcriptomic and metabolomic adaptations from therapy-naïve primary GBM to chemoradiotherapy-resistant recurrent tumors. Our findings revealed unique metabolic transitions that occurred during chemoradiotherapy treatment in GBM cells that were conserved in vitro and in vivo across ten genetically diverse GBM patients. In a pre-clinical animal trial, a novel metabolism-targeting combinatorial treatment regimen prevented the emergence of recurrent tumors and significantly prolonged survival compared to chemoradiotherapy alone. Altogether, our results uncover distinct metabolic adaptations mediating therapy resistance and tumor recurrence in GBM. These findings emphasize the need to consider metabolic flexibility and dependencies for effective therapeutic strategies, offering potential avenues to overcome chemoresistance in recurrent GBM tumors.

Solvent- and Catalyst-Free Environmentally Benign High Hydrostatic Pressure-Assisted Synthesis of Bioactive Hydrazones and the Evaluation of Their Stability Under Various Storage Conditions.

Our group has seen great promise in using substituted diaryl-hydrazones to alleviate oxidative stress in preeclampsia. Specifically, fluorinated diaryl-hydrazones have shown great efficacy, confirmed via antioxidant assays and animal trials using pregnant mice. In addition to efficient antioxidant properties, these diaryl-hydrazones are also considered non-toxic. While the synthesis of these compounds is relatively simple, it commonly utilizes undesirable solvents and glacial acetic acid as the catalyst; additional solvents are needed for the isolation of the desired products, which negatively affects the green synthesis of the hydrazones. To combat this possible industrial roadblock, we have begun incorporating the use of hydrostatic high pressure (HHP) in the synthesis. The use of HHP allowed us to synthesize substituted diaryl-hydrazones in a 1:1 molar ratio without the need for solvents or acid catalysts. The optimized procedure can produce nearly quantitative yields, leading to an easier isolation of the products. Different HHP methodologies, such as constant high-pressure treatment and cycling (with different number of cycles, holding and decompression times) were applied and cycling was observed to be the most efficient activation for the majority of the reactions. Stability experiments were also conducted with one of the products and observed that although the solid-state storage does not alter the hydrazone, storing it in various solvents may significantly decrease the concentration of the active component which should be considered when performing the biochemical/biological assays.

PDGFRB promotes dedifferentiation and pulmonary metastasis through rearrangement of cytoskeleton under hypoxic microenvironment in osteosarcoma

BackgroundOsteosarcoma (OS) cells commonly suffer from hypoxia and dedifferentiation, resulting in poor prognosis. We plan to identify the role of hypoxia on dedifferentiation and the associated cellular signaling. MethodsWe performed sphere formation assays and determined spheroid cells as dedifferentiated cells by detecting stem cell-like markers. RNAi assay was used to explore the relationship between hypoxia inducible factor 1 subunit alpha (HIF1A) and platelet derived growth factor receptor beta (PDGFRB). We obtained PDGFRB knockdown and overexpression cells through lentiviral infection experiments and detected the expression of PDGFRB, p-PDGFRB, focal adhesion kinase (FAK), p-FAK, phosphorylated myosin light chain 2 (p-MLC2), and ras homolog family member A (RhoA) in each group. The effects of PDGFRB on cytoskeleton rearrangement and cell adhesion were explored by immunocytochemistry. Wound-healing experiments, transwell assays, and animal trials were employed to investigate the effect of PDGFRB on OS cell metastasis both in vitro and in vivo. ResultsDedifferentiated OS cells were found to exhibit high expression of HIF1A and PDGFRB, and HIF1A upregulated PDGFRB, subsequently activated RhoA, and increased the phosphorylation of MLC2. PDGFRB also enhanced the phosphorylation of FAK. The OS cell morphology and vinculin distribution were altered by PDGFRB. PDGFRB promoted cell dedifferentiation and had a significant impact on the migration and invasion abilities of OS cells in vitro. In addition, PDGFRB increased pulmonary metastasis of OS cells in vivo. ConclusionOur results demonstrated that HIF1A up-regulated PDGFRB under hypoxic conditions, and PDGFRB regulated the actin cytoskeleton, a process likely linked to the activation of RhoA and the phosphorylation of, thereby promoting OS dedifferentiation and pulmonary metastasis.

Long-term leucine supplementation increases body weight in goats by controlling appetite and muscle protein synthesis under protein-restricted conditions

An inadequate amino acid (AA) supply in animals under protein-restricted conditions can slow skeletal muscle growth. Protein translation can be activated by short-term leucine (Leu) stimulation; however, whether muscle mass increases under long-term Leu supplementation and how the gut and muscle respond to Leu supplementation are largely unknown. In this study, we investigated if muscle mass increases with long-term Leu supplementation under protein-restricted conditions. We identified changes in the link between the gut and muscles under different amino acid supply conditions, using goats as the study object. A total of 27 Xiangdong black male goats with average initial body weight (BW) of 10.88 ± 1.22 kg were randomly divided into three dietary treatments: a normal protein diet (NP, 14.24% crude protein [CP]); a low protein diet (LP, 8.27% CP with supplemental 1.66% rumen-protected lysine [RPLys] and 0.09% rumen-protected methionine [RPMet]); and LP diet with rumen-protected Leu (RPLeu) (LP + RPLeu, 8.75% CP with supplemental 1.66% RPLys, 0.09% RPMet and 1.46% RPLeu). The animal trial lasted for 110 d, consisting of 20 d of adaptation and a 90 d of experimental period. The results showed that long-term protein restriction increased gut tryptophan hydroxylase 1 (TPH1) activity (P < 0.001), tryptophan (Trp) catabolism (P < 0.001), and 5-Hydroxytryptamine (5-HT) synthesis (P < 0.001), which all subsequently reduced goat appetite. Long-term Leu supplementation inhibited 5-HT synthesis (P < 0.001), decreased Trp catabolism in the gut, and increased appetite in goats. Long-term protein restriction enhanced jejunal and ileal branched-chain amino acid transferase (BCAT) (P < 0.001) and branched-chain α-Keto acid dehydrogenase (BCKD) (P = 0.048) activities, which increased branched-chain amino acid (BCAA) catabolism. Immunofluorescence results showed that protein restriction decreased the intestinal mucosal expression of solute carrier family 1 member 5 (SLC1A5) (P = 0.032) and solute carrier family 7 member 5 (SLC7A5) (P < 0.001), reduced BCAA transport from the mucosa to the blood, lowered BCAA levels in the blood (P < 0.001). Western blot results showed that protein restriction inhibited mammalian target of rapamycin (mTOR) pathway activation in goat muscles. Leu supplementation increased BCAA translocation from the intestine to the blood and promoted activation of the muscle mTOR pathway and protein synthesis. In conclusion, our results suggest that Leu supplementation in low-protein diets improves appetite and alleviates the inhibition of muscle protein synthesis in goats.

Development of Biscuits from Winged Beans (Psophocarpus tetragonolobus L.) as a Primary Source for Milk Production

Background: The high prevalence of stunting, notably in Aceh at 31.2%, highlights the urgent requirement for effective nutrition interventions. Utilizing local resources like winged bean flour in food products can play a vital role in enhancing maternal and infant health, addressing malnutrition, and improving breastfeeding outcomes Objective: This study aims to develop biscuits from winged beans (Psophocarpus tetragonolobus L.) as a primary source for enhancing milk production. Winged beans are known to be rich in protein and essential nutrients for the health of breastfeeding mothers. Methodology: This study employs a true experimental design involving clinical trials on rabbits, proximate analysis, and sensory acceptance testing. In the first year, the focus is on developing winged bean biscuit formulas, conducting nutritional and contamination tests, and measuring weight and milk volume in animal trials. The research will take place from March to August 2024 at various locations, including the Nutrition Department at Aceh Health Polytechnic and Syiah Kuala University. In this research, biscuits were produced with varying additions of winged bean flour at 0%, 10%, 15%, and 20% in the biscuit recipe. Results: The results indicate that biscuits containing winged bean flour can increase milk production in female rabbits and show significant weight gain after biscuit consumption. Proximate analysis reveals good nutritional content, while testing for harmful metal contamination ensures product safety. Organoleptic acceptance testing shows that these biscuits are well-received by consumers. These findings affirm the potential of winged beans as a local food source that can support the health of mothers and children, contributing to efforts to combat malnutrition in Indonesia. Conclusion:. Biscuit made from winged bean flour can enhance milk production and weight gain in female rabbits, while offering good nutritional content and being free from contamination. These findings support the potential of winged beans as a local food source to address malnutrition in Indonesia.

Radio-protective effects of melatonin therapy against testicular oxidative stress: a systematic review and meta-analysis of rodent models.

Radiation exposure is a concern in today's world, given the widespread use of electronic devices and medical procedures involving ionizing and non-ionizing radiation. Radiations may cause male infertility by inducing oxidative stress in testicular tissue. Melatonin has antioxidant properties. The authors systematically reviewed the literature for the studies that have investigated the effects of melatonin therapy on radiation-induced oxidative stress in rodents' testicular tissue. PubMed, Scopus, and Web of Science were searched for relevant animal trials. Standardized mean difference and 95% CIs were used to pool the data. Subgroup and sensitivity analyses were done. The risk of bias was assessed using SYRCLE tool. Outcomes: histopathology and sperm analyses (testicular apoptotic cells, Johnsen's testicular biopsy score, seminiferous epithelial height, tubular diameter, sperm motility, viability, count, and morphology, concentration of spermatid, spermatocyte, and spermatogonia), body and testes weights (absolute and relative body and testicular weights), reproductive hormones (serum prolactin, FSH, and testosterone), and oxidative stress tissue markers (TBARS, CAT, GSH, GSH-Px, MDA, SOD, and XO, and total antioxidant capacity). Rats and mice were exposed to electromagnetic radiations (gamma, roentgen, microwave, radiofrequency, and high-power line energy) and particle waves (radioiodine and carbon-ion). Melatonin therapy was significantly associated with improved male reproduction. Radiation exposure harms male fertility, but melatonin, as an antioxidant, is potentially associated with improved male reproductive function in rodents. Inconsistencies in research require further investigations.

Dietary supplementary with ellagic acid improves the intestinal barrier function and flora structure of broiler chicken challenged with E. coli K88

Ellagic acid (EA) contributes to the immunity and anti-oxidant function of body, whereas there are few reports about its effect on the intestinal health and growth performance of broiler chickens. Hence, the present study was arranged to investigate the effect of dietary supplementary with EA on the intestinal barrier function and flora structure of broiler chickens challenged with Escherichia coli K88 (E. coli K88). A total of 216 healthy 1-day-old, Ross 308 broilers with uniform weight were randomly assigned into three treatment groups, six replicates in each group and twelve birds in each replicate. Broilers in the control (CTR) group and E. coli K88 infected group (ETEC) were fed with the basic diet, and 200 mg/kg EA was supplemented into the diet of the E. coli K88 infected group treated with EA (EAETEC). The animal trial had lasted for 42 days, and the outcomes showed that the ADG and ADFI during the animal trial, the jejunal villi height (VH) and the ratio of VH to crypt depth (CD) tended to be decreased with E. coli K88 treated (P< 0.05). Additionally, the level of serum diamine oxidase (DAO) and intestinal malondialdehyde (MDA) were elevated, the activity of intestinal total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px), the mRNA levels in jejunal claudin-1 and occludin were down-regulated with E. coli K88 treated as well as the transcription levels of ileal Mucin-2, aquaporin-3 (AQP-3) and Na+/H+ exchanger proteins-3 (NHE-3) (P< 0.05). In addition, E. coli K88 down-regulated the α-diversity index of cecal flora, the ratio of Bacteroidota to Firmicutes and the relative abundance of Barnesiella were up-regulated and it of Alistipes was descended with E. coli K88 treated (P< 0.05). Beyond that, the content of propionic acid in the cecal chyme was decreased and the amino acid metabolic pathways were inhibited with E. coli K88 challenged (P< 0.05). Additionally, there was a significant positive correlation between the relative abundance of Alistipes and the levels of butyric acid in the caecal chyme and the activity of GSH-Px in the intestine (P< 0.05). Interestingly, dietary supplementary with EA could reshape the intestinal flora structure and alleviate the above negative effects of E. coli K88 on broiler chickens. In conclusion, dietary supplementary with ellagic acid improved the intestinal barrier function and flora structure of broiler chickens challenged with E. coli K88.