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Context: Cardiovascular disease is a leading cause of mortality worldwide. Following acute myocardial infarction (AMI), cardiac cells release many cytokines and proteolytic enzymes that regulate the extracellular matrix (ECM), including matrix metalloproteinases (MMPs), tissue inhibitor of metalloproteinases (TIMPs), and phosphorylated microtubule-associated protein 4 (Phospho/pMAP4). Aims: To analyze the role of colchicine in ECM regulation post-ischemic in cell line culture towards MMPs, TIMPs, and pMAP4. Methods: This in vitro experimental study was conducted on the 3T3 fibroblast cell line subjected to hypoxic conditions (CoCl2 treatment) and compared with 3T3 cells experiencing hypoxia/ischemic and colchicine treatment. MMP-1, TIMP-1, and pMAP4 expression were measured using flow cytometry. Data analysis was performed using One-Way ANOVA with a 95% confidence level. Results: Treatment with colchicine in the 3T3 fibroblast cell culture regulated the extracellular matrix by increasing TIMP-1 expression and reducing MMP-1 and pMAP4 expression (p

Background: Chronic physical stress has many effects on the nervous system and can cause structural changes in different parts of the brain and hemomodulatory, including hormonal. Current pharmacotherapeutic treatments have limited efficacy and are associated with many deleterious side effects Aim: The aim of this research is to determine how apis dorsata forest honey administration affects FSH and LH levels in rats who are subjected to forced swim tests as a model of chronic physical stress placed in a container filled with water from which it cannot escape. Methods: This was an experimental laboratory study with 32 rats divided into four treatment groups: control (C), Treatment 1 (T1) with a forced swim test + honey (2 g/rat/day), Treatment 2 (T2) with a forced swim test + honey (4 g/rat/day), and Treatment 3 (T3) with a forced swim test + honey (6 g/rat/day). All treatments were administered for 14 days. Then blood was taken for FSH and LH serum tests, and a one-way ANOVA and Duncan test were used to statistically test the data analysis. Results: The results of this study indicate that the administration of forest honey had no significant effect (p > 0.05) on the FSH parameter, but there was a significant decrease in LH levels in the T2 and T3 groups (p [Open Vet J 2024; 14(2.000): 738-742]

Background There is an essential need for cardiomyocyte regeneration among patients with heart failure. Transplantation of dedifferentiated fat (DFAT) cells may lead to an improvement of cardiomyocyte regeneration among heart failure patients. We believe that DFAT cells are promising candidate cell sources for cardiac regeneration. However, the pathway underlying how DFAT cells of the adipose lineage differentiate into mature cardiomyocytes isn’t fully understood. Methods We conducted an experimental laboratory study on isolated DFAT cells from adipose tissue of healthy adults. Then, we treated cells with different concentrations of reversine (10, 20 and 40 nM), and performed RNA extraction and cDNA synthesis. Next, we used a ceiling culture method based on the buoyancy properties of mature lipid-filled adipocytes. Stemness expression (Octamer-binding transcription factor 4 [Oct4], brachyury, Fetal liver kinase 1 [Flk-1]) was quantified by reverse transcription-quantitative (RT-q)PCR, while cardiomyocyte expression (Transcription factor GATA-4 [GATA4] and cardiac troponin T [cTnT]) was quantified by immunocytochemistry. Results ANOVA with Tukey’s post-hoc found that 10 nM reversine increased greater Flk-1 expression compared to the control group (MD: 5.037 + 0.998; p < 0.001), but there were no significant changes among Oct4 (MD: 0.013 + 1.244; p = 0.99) and brachyury expression (MD: 0.157 + 0.084; p = 0.252). Kruskal-Wallis revealed that the expression of GATA4 (1.65 [0.41-1.98] to 0.015 [0.007-0.034]; p =0.017) reduced significantly from day 7 until day 21 and cTnT (5.07 [6.62-8.91] to 8.22 [6.81-9.40]; p= 0.001) increased significantly from day 7 until day 21. Conclusions Reversine could increase the expression of Flk-1, but it was unable to stimulate the expression of Oct4 and brachyury related to cell stemness. An optimal concentration of 10 nM reversine may have the greatest effect on enhancing the differentiation of DFAT cells into mature cardiomyocytes, as indicated by higher cTnT expression between cells.

Background: Inflammation plays a critical role in myocardial infarction as a critical process in the development of heart failure, involving the development of cardiac fibrosis. Colchicine is a well-established anti-inflammatory drug, but its scientific application in controlling post-acute myocardial infarction (AMI) inflammatory processes has not been established. IL-10 is a key cytokine in modulating inflammatory responses, underscoring its potential as a crucial therapeutic target of colchicine. The objective was to explore the protective role of IL-10 modulated by colchicine in myocardial healing and repair following AMI, particularly cardiac fibrosis. Methods: The predicted protein of colchicine was assessed using WAY2DRUG PASS as probability active value. Proteins associated with colchicine, cardiac fibrosis, and acute myocardial infarction were analyzed with DisGeNET and Open Target databases. Analysis and visualization of protein–protein interactions were conducted using STRING and Cytoscape. A 3T3 cell line treated with CoCl2 was used to mimic hypoxic. HIF-1α and IL-10 expression were measured by flow cytometry and analyzed using a one-way ANOVA test. This observational clinical trial examined acute myocardial infarction patients undergoing immediate and delayed primary percutaneous coronary interventions. Subjects were randomized into control groups receiving placebo and intervention groups treated with colchicine. Assessments occurred at 24 h and five days after the intervention. IL-10 expression in the clinical trial was measured by ELISA and analyzed using a T-test. Results: Colchicine demonstrates promising bioactivity in treating acute myocardial infarction, with notably activity values highlighting its probable role as a tubulin antagonist (0.744), beta-tubulin antagonist (0.673), and NOS2 inhibitor (0.529). Its primary action targets IL-10, with the protein–protein interactions analysis indicating interactions between IL-10 and key inflammatory mediators—IL-1β, IFN-γ, CCL2, TNF, and TGF-β1—during acute myocardial infarction and cardiac fibrosis. Hypoxic conditions in the CoCl2-induced 3T3 cell model show significantly elevated HIF-1α compared to controls (p < 0.0001). Colchicine use significantly increased IL-10 expression in CoCl2-treated cells (p < 0.0001) and in AMI patients within five days (p < 0.05). Conclusions: Colchicine may bolster the anti-inflammatory response post-myocardial infarction by activating IL-10 pathways in fibroblasts and in clinical settings, potentially reducing inflammation after AMI. Further investigation into broader aspects of this pathway, particularly in cardiac fibroblasts, is required.

During storage at low temperatures, the spermatozoa quality changes due to cold shock and free radicals. Diluent supplementation with antioxidants is an effort to maintain the quality of spermatozoa during storage. This study aimed to evaluate the α-tocopherol effect in CEP extender on sperm quality, membrane integrity, and lipid peroxidation during storage at 4°C-5°C. This was a laboratory experiment that compared the use of 2 mM of α-tocopherol in CEP with no addition of α-tocopherol (as control) in five bulls. Semen was collected from Brahman bulls, diluted in CEP with and without α-tocopherol, and stored at 4°C-5°C. Sperm motility and viability were investigated by a light microscope at a ×400 magnification using Eosin-Nigrosin staining. Moreover, membrane integrity was evaluated by lipid peroxidation using the MDA assay and hypoosmotic swelling test. The sperm motility, viability, and membrane integrity were higher in CEP with α-tocopherol. Lipid peroxidation was significantly different between the treatment and control groups. The α-tocopherol supplementation in the diluent CEP could maintain the spermatozoa quality during storage at 4°C-5°C.

Background: Toxoplasma gondii is one of the zoonotic protozoa parasites. It can prevalently infect humans and warm-blooded animals, causing human health problems and substantial economic losses to the livestock industry worldwide. Chicken is one of the potential sources of toxoplasmosis, but there is no report of the prevalence of toxoplasmosis and their genotypes in free-range chickens in Libya. Aim: The current study aims to conduct a survey of molecular prevalence and identify the Toxoplasma gondii genotype in free-range chickens and its association with the risk factors of age, gender, and region in Northeastern Libya. Methods: This study was conducted by examining a total of 315 free-range chicken organs (brain and heart) derived from three administrative districts in Northeastern Libya. The molecular prevalence was determined by PCR technique using B1 gene amplification and the Toxoplasma gondii genotype was determined by nested PCR-RFLP of GRA6 gene amplicon with restriction enzymes (MseI). Results: The overall molecular prevalence of Toxoplasma gondii in free-range chicken in all three districts was 9.5% (30/315), and the highest (15.4%) was in the Al-Marj district (p =0.01; ?2 =9.238). The highest prevalence of Toxoplasma gondii by age was in chickens aged more than two years (p = 0.001; ?2 = 15.530). The difference in Toxoplasma gondii prevalence in male and female chickens was not significant (p = 0.372; ?2 = 0.798). The predominant genotype I (93.3%) had identified at position 544 bp and 194 bp at the GRA6 marker, and only two positives were from genotype II (6.7%) at 700 bp and 100 bp fragment. Conclusion: The molecular prevalence of toxoplasmosis in free-range chicken in three districts in Northeastern Libya was 9.5%, and the highest rate was shown in the Al Marj district. Chicken by age more than two years had more risk to transmit toxoplasmosis in human. There was no different infection risk by consuming male or female free-range chicken. It is the first report to determine the predominant genotype, which was genotype I. [Open Vet J 2023; 13(2.000): 225-232]

Background and Aim: Zebrafish are frequently used as model organisms in scientific research as their genes mirror those of humans. Aeromonas hydrophila bacteria can infect humans and animals, mainly fish. This study aimed to identify the concentration and route of A. hydrophila infection in adult zebrafish. Zebrafish had been used as a challenge test by analyzing their hematological profiles, blood glucose levels, and survival rates. Materials and Methods: Induction of cell supernatant free (CSF) from A. hydrophila bacteria in adult zebrafish was carried out via bath immersion (BI), intraperitoneal injection (IPI), intramuscular injection (IMI), and healthy zebrafish as a control (C). The bacterial concentrations were 107, 109, and 1011 colony-forming units (CFU)/mL. At 24 h post-infection, the outcomes of infection were evaluated based on survival rates, hematological profiles, and blood glucose levels. A one-way analysis of variance with a confidence level of 95% was employed to examine the data. Results: In the BI, IPI, and IMI treatment groups, the survival rate of the fish reached a peak of 100%, 22%–100%, and 16%–63%, respectively, compared with the injection technique. In the IMI2 group, a 109 CFU/mL bacterial concentration was determined to correspond to the lethal dosage 50. All infection groups had lower erythrocyte and hemoglobin counts but higher leukocyte counts than the control group. The blood sugar levels of the healthy and infected groups were not significantly different. Conclusion: The route of A. hydrophila infection through Intramuscular injection with a concentration of 109 CFU/mL indicated a high performance compared to other techniques. This method could be developed as a reproducible challenge test.

Background: Staphylococcus and Aeromonas bacteria are pathogens in humans and animals. The therapy disrupts the virulence structure of the bacteria, resulting in bacterial death. Currently, chemical drugs have resulted in many resistant bacteria, so it is necessary to find alternative natural materials that are not toxic and do not quickly induce resistance. Aims: This study aimed to analyze the potential of methanol extract from BSF prepupa as an antibacterial agent against S. aureus and Aeromonas through in silico and in vitro tests. Methods: The BSF prepupae methanol extract was then analyzed for protein and fatty acid contents. Molecular docking of the active ingredients (defensin, chitin, and chitosan as well as fatty acids) in BSF was downloaded from the NCBI database and docked with the Hex Cuda 8.0 program, Correlation type parameters Shape + Electro and Grid Dimension 0.6. Docking results were analyzed using the Discovery Studio program version 21.1.1. Results: The highest fatty acid contents in the extract were palmitic acid and myristic acid. Methanol extract from BSF prepupae acted as a bactericidal agent against S. aureus at a concentration of 320 mg/ml, in contrast to Aeromonas, which still showed bacterial growth. The results of the in silico test showed that defensin–aerolysin and defensin–hemolysin were bound to the same active site area. However, the amount of binding energy produced by 69-Defensin-83-aerolysin was higher than all defensin types in BSF against Aeromonas. Chitin and chitosan showed a bond on the active site of aerolysin and hemolysin, but chitosan had a stronger bond than chitin. In silico study also showed the strongest binding affinity of BSF fatty acids to isoleucyl-tRNA synthetase of S. aureus. Conclusion: The study showed that methanol extract from BSF prepupae had more potential as an antibacterial agent against S. aureus than Aeromonas in vitro and in silico. [Open Vet J 2023; 13(1.000): 48-63]

A decoction of Rhizophora mucronata Lam. mangrove bark is used as an antidiabetic treatment in Asia. Kombucha tea is a fermented beverage, which is also claimed to be antidiabetic. In this work, the potency of R. mucronata leaves as α-glucosidase inhibitor was studied to assess whether it could be a suitable alternative to the use of R. mucronata bark. α-glucosidase inhibitory activities were determined for three extracts prepared from R. mucronata leaves, being the unfermented tea of R. mucronata leaves, the fermented kombucha tea and an 80% methanolic extract of the residual R. mucronata leaves. Flavonoid glycosides were identified in tea powder, kombucha tea and in the crude methanolic extract. Both the unfermented tea and the kombucha tea after 7 days of fermentation inhibited α-glucosidase with IC50 values of 0.12 ± 0.02 mg/mL and 0.09 ± 0.04 mg/mL, respectively. The methanolic extract showed a stronger α-glucosidase inhibitory activity compared to the kombucha tea and tea powder with an IC50 value of 0.0435 ± 0.0007 mg/mL. Acarbose, used as a positive control, inhibited α-glucosidase with an IC50 value of 2.4 ± 0.2 mg/mL. It was found that the three types of preparations of R. mucronata all were potent α-glucosidase inhibitors.

Diabetes mellitus causes a pancreatic β-cells damage due to the increase of oxidative stress, thereby reducing Superoxide Dismutase 1 (SOD1) activity. This enzyme activity can be increased by utilizing antioxidant compounds from the avocado seed extract. This study aims to determine the increase of SOD1 expression and the repair of pancreatic β-cells tretaed with the ethanol extract of avocado seeds (Persea americana Mill). It is also aimed to observe the expression of pancreatic SOD1 and its histopathology change. The expression was measured by using immunohistochemical. This research used 20 Wistar rats aged of 2-3 moths, weighing of 150-200 grams. The diabetic rats were induced by streptozotocin (STZ) with dose of 30 mg/kg BW intraperitoneally. The rats were divided into 5 groups, namely: group 1 is healthy rats, group 2 is diabetic rats, while the group 3, 4 and 5 are therapeutic groups with 300, 350 and 400 mg/kg BW doses of ethanol extract for 14 days treatment. The results showed the ethanolic extract of avocado seeds has a potential as an antidiabetic agents based on the increase of pancreatic SOD1 (p

This study was aimed to characterize lignin peroxidase (LiP) obtained from Phanerochaete chrysosporium ITB isolate. The characterizations included molecular weight, the pH and optimum working temperature of the crude extract of the enzyme, the temperature stability, the effect of metal ions and inhibitors, their precipitation with ethanol, and the storage stability. The LiP of P. chrysosporium ITB isolates was 34 kDa. The crude extract of LiP displays high activity at pH between 3 until 5 and 26–32 °C, has good thermal stability at 26–32 °C for 20 h. The activity is affected by Pb2+, K+, Co2+, Fe2+, Cd2+, Mg2+, and Cu2+ EDTA, Na+, Cr3+, Hg2+, NaN3, Ni2+, and Ca2+ ions, is not affected by Mn2+ and Zn2+ ions, precipitated with the optimum ethanol at 64% ethanol saturation which results in an increase in specific activity of 2.3 times. The crude extract storage at 0 °C is more stable than the precipitate resulting from ethanol precipitation and resuspension from ethanol precipitation. These results strengthen that LiP from P. chrysosporium is another LiP isoenzyme that can be used for bioremediation processes. Unfortunately, the concentration using the ethanol precipitation method has not been effective, so further studies using other methods should be required.

Coconut water is part of the coconut fruit, that has bioactive compounds that are thought to own high inhibitor activity to the presence of tannin compounds. However, this compound is antinutritional because it can interfere with macromolecule out there within the body. Considering chance of tannins, these parts are often reduced by fermentation techniques using lactic acid bacteria. This analysis aims to review the addition of dragon fruit and lemon peels in fermentation coconut water using lactic acid bacteria in order to reduce tannin levels and its potential capability as antioxidants. Coconut water fermented drink was prepared by fermentation technique using lactic acid bacteria with the addition of variations in mass of dragon fruit peel and lemon peel consisting of 5 treatments. The optimum conditions for fermentation of coconut water were determined from the decrease in tannin levels, total acid, and the highest total lactic acid bacteria. The optimum conditions for fermentation of coconut water were then tested using an HPLC instrument. Finally, the antioxidantactivity was tested using the FRAP (Ferric Reducing Antioxidant Power) method. The results showed that the addition of dragon fruit and lemon peels in fermented coconut water can reduce tannin levels, can increase total acid levels and the number of lactic acid bacteria formed in coconut water fermented drinks. The best proportion was obtained in the treatment of rasio of mass 1:2 resulted in the highest decrease in tannin content of 83.94 mg/100 g followed by a total acid value of 1.13% and the number of lactic acid bacteria was 8.4 × 107 CFU/mL. The HPLC chromatogram showed that fermented coconut water using lactic acid bacteria contained organic acid compounds such as lactic acid, acetic acid, and ascorbic acid. The highest antioxidant activity test was 82.37 mmgAAE/g. This study confirmed that the addition of dragon fruit and lemon peels affected the degradation of tannins and increased antioxidant activity during coconut water fermentation using lactic acid bacteria.

Saponin-rich sea cucumber extracts have shown antidiabetic effects in a few reports. Although the triterpene glycosides of sea cucumbers are commonly isolated from their Cuvierian tubules, these are absent in Holothuria atra Jaeger. Therefore, this study intended to investigate the saponin profile in the body wall of H. atra, as well as to assess the α-glucosidase inhibitory activity of the H. atra extracts. The chemical profiling of sea cucumber extracts was conducted by UPLC-HRMS analysis. This resulted in the tentative identification of 11 compounds, 7 of which have not been reported in the H. Atra body wall before. Additionally, two triterpene glycosides were purified and their structures were elucidated based on HRMS and NMR data: desholothurin B (1), and a novel epimer, 12-epi-desholothurin B (2). Moreover, the fatty acid profile of the H. atra body wall was investigated by GC-MS. It was found that the Me90 fraction of the H. atra body wall showed the strongest α-glucosidase inhibitory activity (IC50 value 0.158 ± 0.002 mg/mL), thus making it more potent than acarbose (IC50 value 2.340 ± 0.044 mg/mL).

Background: Diabetes mellitus (DM) is a chronic and costly disease that has become a primary concern worldwide. Type 1 diabetes mellitus is categorized as an autoimmune disease, which results in islet cell apoptosis and insulin-dependent. GAD65 is known as a potential marker of impaired pancreatic β cell function that appears in the initial phase of type 1 DM and latent autoimmune diabetes in adults (LADA). This study aimed to develop a novel rapid test of anti-GAD65 autoantibodies in human serum samples. Methods: We have developed a rapid test for anti-GAD65 autoantibodies in this assay based on the reverse-flow immunochromatography method. Human recombinant-protein antigen for GAD65 was attached as the control line over the nitrocellulose membrane. On the other side, the goat anti-mouse immunoglobulin G (IgG) was coated on the same membrane as a control line. The positive result for GAD65 was confirmed by a colloidal gold signal on the strip. Our novel assay analyzed 276 healthy subjects and 51 type 1 diabetes individuals serum samples from several ethnicities in Indonesia for this study. Results: Among the 276 healthy samples, 225 samples were identified as positive for anti-GAD65 autoantibodies, while 51 samples were negative. Interestingly, the positive results for anti-GAD65 autoantibodies were linear to the decreasing of high-density lipoprotein (HDL) levels and inversely associated with triglyceride levels. A significant correlation with age was observed in all groups. The sensitivity and specificity test proved that this kit has higher accuracy (AUC value = 0.960). Conclusion: The significant advantages of our rapid test for anti-GAD65 autoantibodies provide higher sensitivity, specificity, and stability compared to previous commercial kits. Therefore, it could be proposed as the future clinical diagnostic kit for patient management of type 1 DM.

Background There is an essential need for cardiomyocyte regeneration among patients with heart failure. Transplantation of dedifferentiated fat (DFAT) cells may lead to an improvement of cardiomyocyte regeneration among heart failure patients. We believe that DFAT cells are promising candidate cell sources for cardiac regeneration. However, the pathway underlying how DFAT cells of the adipose lineage differentiate into mature cardiomyocytes isn’t fully understood. Methods We conducted an experimental laboratory study on isolated DFAT cells from adipose tissue of healthy adults. Then, we treated cells with different concentrations of reversine (10, 20 and 40 nM), and performed RNA extraction and cDNA synthesis. Next, we used a ceiling culture method based on the buoyancy properties of mature lipid-filled adipocytes. Stemness expression (Octamer-binding transcription factor 4 [Oct4], brachyury, Fetal liver kinase 1 [Flk-1]) was quantified by reverse transcription-quantitative (RT-q)PCR, while cardiomyocyte expression (Transcription factor GATA-4 [GATA4] and cardiac troponin T [cTnT]) was quantified by immunocytochemistry. Results ANOVA with Tukey’s post-hoc found that 10 nM reversine increased greater Flk-1 expression compared to the control group (MD: 5.037 + 0.998; p < 0.001), but there were no significant changes among Oct4 (MD: 0.013 + 1.244; p = 0.99) and brachyury expression (MD: 0.157 + 0.084; p = 0.252). Kruskal-Wallis revealed that the expression of GATA4 (1.65 [0.41-1.98] to 0.015 [0.007-0.034]; p =0.017) reduced significantly from day 7 until day 21 and cTnT (5.07 [6.62-8.91] to 8.22 [6.81-9.40]; p= 0 .001) increased significantly from day 7 until day 21. Conclusions Reversine could increase the expression of Flk-1, but it was unable to stimulate the expression of Oct4 and brachyury related to cell stemness. An optimal concentration of 10 nM reversine may have the greatest effect on enhancing the differentiation of DFAT cells into mature cardiomyocytes, as indicated by higher cTnT expression between cells.

Background There is an essential need for cardiomyocyte regeneration among patients with heart failure. Transplantation of dedifferentiated fat (DFAT) cells may lead to an improvement of cardiomyocyte regeneration among heart failure patients. We believe that DFAT cells are promising candidate cell sources for cardiac regeneration. However, the pathway underlying how DFAT cells of the adipose lineage differentiate into mature cardiomyocytes isn’t fully understood. Methods We conducted an experimental laboratory study on isolated DFAT cells from adipose tissue of healthy adults. Then, we treated cells with different concentrations of reversine (10, 20 and 40 nM), and performed RNA extraction and cDNA synthesis. Next, we used a ceiling culture method based on the buoyancy properties of mature lipid-filled adipocytes. Stemness expression (Octamer-binding transcription factor 4 [Oct4], brachyury, Fetal liver kinase 1 [Flk-1]) was quantified by reverse transcription-quantitative (RT-q)PCR, while cardiomyocyte expression (Transcription factor GATA-4 [GATA4] and cardiac troponin T [cTnT]) was quantified by immunocytochemistry. Results ANOVA with Tukey’s post-hoc found that 10 nM reversine increased greater Flk-1 expression compared to the control group (MD: 5.037 + 0.998; p < 0.001), but there were no significant changes among Oct4 (MD: 0.013 + 1.244; p = 0.99) and brachyury expression (MD: 0.157 + 0.084; p = 0.252). Kruskal-Wallis revealed that the expression of GATA4 (1.65 [0.41-1.98] to 0.015 [0.007-0.034]; p =0.017) reduced significantly from day 7 until day 21 and cTnT (5.07 [6.62-8.91] to 8.22 [6.81-9.40]; p= 0 .001) increased significantly from day 7 until day 21. Conclusions Reversine could increase the expression of Flk-1, but it was unable to stimulate the expression of Oct4 and brachyury related to stem cell-ness. An optimal concentration of 10 nM reversine may have the greatest effect on enhancing the differentiation of DFAT cells into mature cardiomyocytes, as indicated by higher cTnT expression between cells.

Background: Neurodegeneration due to neurotoxicity is one of the phenomena in temporal lobe epilepsy. Experimentally, hippocampal excitotoxicity process can occur due to kainic acid exposure, especially in the CA3 area. Neuronal death, astrocyte reactivity and increased calcium also occur in hippocampal excitotoxicity, but few studies have investigated immediate effect after kainic acid exposure. The organotypic hippocampal slice culture (OHSC) is a useful model for studying the neurodegeneration process, but there are still many protocol differences. In this study, minor modifications were made in the OHSC protocol. Methods: OHSC was obtained from two healthy wild type Wistar rats aged P10. Healthy culture slices were obtained and lasted up to 10 days in vitro (DIV 10). Bath application of kainic acid for 48 hours in DIV 10 followed by observation of its initial effects on neurons, astrocytes, and calcium via the expression of MAP2, GFAP, and intracellular calcium, subsequently. Results: After 48 h of kainic acid administration, there was a significant increase in intracellular calcium (p = 0.006 < α), accompanied by a significant decrease in MAP2 (p = 0.003 < α ) and GFAP (p = 0.010 < α) expression. Conclusion: These findings suggest early neuronal and astrocyte damage at the initial onset of hippocampal injury. This implies that astrocyte damage occurs early before an increase in GFAP that characterizes reactive astrogliosis found in other studies. Damage to neurons and astrocytes may be associated with increased intracellular calcium. It is necessary to develop further research regarding expression of calcium, MAP2, and GFAP at a spatial time after exposure to kainic acid and strategies to reduce damage caused by excitotoxicity.

This study aims to detect Escherichia coli which encodes beta-lactamase Cefotaxime (blaCTX-M) gene from the reproductive tract of Bali cattle with repeat breeder cases. This research was conducted from June to August 2021 using 16 Bali cattle with repeat breeder cases. The reproductive fluids were taken using a plastic sheet gun which was inserted into a Brain Heart Infusion medium, isolated in eosin-methylene blue agar (EMB) and identified using biochemical tests. Antibiotic susceptibility testing of E. coli was carried out using the disc diffusion method. Double-disk approximation test was used to screen the presence of E. coli which produces Extended-spectrum beta-lactamase. The polymerase chain reaction (PCR) method was used to detect the blaCTX-M gene of E. coli and sequences of the blaCTX-M gene were phylogenetically analyzed. The research results obtained three E. coli isolates from 16 reproductive tract fluids of Bali cattle. Antibiotic sensitivity tests showed that 100% of E. coli was resistant to penicillin G and oxytetracycline. 66.66% of E. coli was resistant to cefotaxime (CTX) and gentamicin, and 33.33% of E. coli was resistant to tetracycline. Escherichia coli isolates that were resistant to penicillin and CTX showed positive results in the double-disk approximation test. The results of E. coli detection using PCR showed that three E. coli isolates encoded the blaCTX-M gene located at 370 bp on gel electrophoresis. The results of the phylogenetic analysis showed that E. coli from the reproductive tract of Bali cattle was related to E. coli that encoded blaCTX-M-14 isolated from humans. [ABSTRACT FROM AUTHOR]

Background: Neurodegeneration due to neurotoxicity is one of the phenomena in temporal lobe epilepsy. Experimentally, hippocampal excitotoxicity process can occur due to kainic acid exposure, especially in the CA3 area. Neuronal death, astrocyte reactivity and increased calcium also occur in hippocampal excitotoxicity, but few studies have investigated immediate effect after kainic acid exposure. The organotypic hippocampal slice culture (OHSC) is a useful model for studying the neurodegeneration process, but there are still many protocol differences. In this study, minor modifications were made in the OHSC protocol. Methods: OHSC was obtained from two healthy wild type Wistar rats aged P10. Healthy culture slices were obtained and lasted up to 10 days in vitro (DIV 10). Bath application of kainic acid for 48 hours in DIV 10 followed by observation of its initial effects on neurons, astrocytes, and calcium via the expression of MAP2, GFAP, and intracellular calcium, subsequently. Results: After 48 h of kainic acid administration, there was a significant increase in intracellular calcium (p = 0.006 < α), accompanied by a significant decrease in MAP2 (p = 0.003 < α ) and GFAP (p = 0.010 < α) expression. Conclusion: These findings suggest early neuronal and astrocyte damage at the initial onset of hippocampal injury. This implies that astrocyte damage occurs early before an increase in GFAP that characterizes reactive astrogliosis found in other studies. Damage to neurons and astrocytes may be associated with increased intracellular calcium. It is necessary to develop further research regarding expression of calcium, MAP2, and GFAP at a spatial time after exposure to kainic acid and strategies to reduce damage caused by excitotoxicity.

Purpose: To investigate casein's role in the expressions of human beta-defensin (HBD)-1, T helper subset (Th)-17, and interleukin (IL)-22 in preterm Rattus norvegicus (RNs). Methods: Twenty-four healthy, first-time pregnant RNs were divided into three groups (G). Group 1 (G1) received regular casein (R) 0.4 g/RN/day, while G2 and G3 received casein supplementation (S) 2 g/RN/day. Each RN was given 10 g of food and drank 10 mL. On day 10, a single intraperitoneal lipopolysaccharide (LPS) was injected with 100 g/kg body weight. Lipopolysaccharide was injected into G3 as a premature model with chorioamnionitis. Histopathological examination was carried out to measure the expression of HBD-1, Th-17, and IL-22 using methyl green counterstaining. Chorioamnion specimens were taken and fixed in 4 % paraformaldehyde, and then evaluated by immunohistochemical (IHC) procedures to obtain immunoreactive score (IMS). Results: The IMS expression of HBD-1 in G1 was 1.187 ± 0.372, while G3 was 9.562 ± 2.351 (p < 0.05). Th-17 expression were G1: 2.062 ± 0.140, G2: 6.122 ± 1.347, and G3: 9.112 ± 2.019 (p < 0.05). Furthermore, IL-22 expressions were G1: 1.825 ± 0.468, G2: 4.700 ± 1.614, and G3: 7.800 ± 1.669 (p < 0.05). Conclusion: Casein supplementation increases HBD-1, Th-17, and IL-22 expression in preterm RN and in the LPS-induced RN chorioamnionitis model. The increase in HBD-1, Th-17, and IL-22 indicate that casein plays a role in the protection and control of chorioamnionitis tissue inflammation. [ABSTRACT FROM AUTHOR]

Background: Parkinson's disease (PD) is the most common chronic progressive neurodegenerative disorder in the older population. In this work, we have developed a formulation of orally disintegrating tablets (ODTs) containing Centella asiatica (CA) encapsulated solid lipid nanoparticles (SLNs) with rapid disintegration and dissolution, thereby providing greater convenience and ease of use to older patients with PD or dysphagia. Methods: The absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties of CA compounds were evaluated using QikProp module of Schrödinger. CA-SLNs were prepared using hot homogenization method. The physicochemical properties and quality control of the pre-compressed powder were characterized. The direct compression method was used to prepare ODTs, and post-compression physical properties were evaluated. Results: In silico study of ADMET properties revealed the CA compounds can follow the criteria for an orally active drug and are within the standard range in terms of 'Rule of Five' and 'Rule of Three”. The characteristics of CA-SLNs developed in a lipid-based nanocarrier showed monodispersed particles with an average particle size of about 37.91±1.55 nm, zeta potential of -10.27±1.37 mV, encapsulation efficiency, and loading capacity of 95.07±1.14%, and 3.99±0.06%, respectively. The results obtained for the pre-compression characterization showed that the CA-SLNs powder mixture had excellent flowability properties and compressibility. Furthermore, these results affected the physical properties of CA-SLNs ODTs with a disintegration time of 14.5s, the acceptance value of content uniformity was 3.2%, and the in vitro dissolution test fulfilled the tolerance limits recommended in the United States Pharmacopeia (USP) monograph. Conclusions: Overall, these results suggest that CA-SLNs ODTs developed with lipid-based nanocarriers can be considered an alternative delivery system to protect the active compound from instability while enhancing permeability through the blood brain barrier (BBB) and can be used in the management of PD in older patients with dysphagia.

Background: Inflammation plays a critical role in myocardial infarction as a critical process in the development of heart failure, involving the development of cardiac fibrosis. Colchicine is a well-established anti-inflammatory drug, but its scientific application in controlling post-acute myocardial infarction (AMI) inflammatory processes has not been established. IL-10 is a key cytokine in modulating inflammatory responses, underscoring its potential as a crucial therapeutic target of colchicine. The objective was to explore the protective role of IL-10 modulated by colchicine in myocardial healing and repair following AMI, particularly cardiac fibrosis. Methods: The predicted protein of colchicine was assessed using WAY2DRUG PASS as probability active value. Proteins associated with colchicine, cardiac fibrosis, and acute myocardial infarction were analyzed with DisGeNET and Open Target databases. Analysis and visualization of protein–protein interactions were conducted using STRING and Cytoscape. A 3T3 cell line treated with CoCl2 was used to mimic hypoxic. HIF-1α and IL-10 expression were measured by flow cytometry and analyzed using a one-way ANOVA test. This observational clinical trial examined acute myocardial infarction patients undergoing immediate and delayed primary percutaneous coronary interventions. Subjects were randomized into control groups receiving placebo and intervention groups treated with colchicine. Assessments occurred at 24 h and five days after the intervention. IL-10 expression in the clinical trial was measured by ELISA and analyzed using a T-test. Results: Colchicine demonstrates promising bioactivity in treating acute myocardial infarction, with notably activity values highlighting its probable role as a tubulin antagonist (0.744), beta-tubulin antagonist (0.673), and NOS2 inhibitor (0.529). Its primary action targets IL-10, with the protein–protein interactions analysis indicating interactions between IL-10 and key inflammatory mediators—IL-1β, IFN-γ, CCL2, TNF, and TGF-β1—during acute myocardial infarction and cardiac fibrosis. Hypoxic conditions in the CoCl2-induced 3T3 cell model show significantly elevated HIF-1α compared to controls (p < 0.0001). Colchicine use significantly increased IL-10 expression in CoCl2-treated cells (p < 0.0001) and in AMI patients within five days (p < 0.05). Conclusions: Colchicine may bolster the anti-inflammatory response post-myocardial infarction by activating IL-10 pathways in fibroblasts and in clinical settings, potentially reducing inflammation after AMI. Further investigation into broader aspects of this pathway, particularly in cardiac fibroblasts, is required. [ABSTRACT FROM AUTHOR]

Introduction: Diabetic kidney disease (DKD), as a diabetes mellitus type 2 (DMT2) complications, is getting more prevalent nowadays. Inflammation is one of the renal injury mechanisms evaluated through the surge in in TNF-α and NF-κβ expression. Impaired expression of gluten transporter 1 (GLUT1) and GLUT2 reduces glucose uptake. DBLS3233 is a novel anti-diabetes agent and Indonesian herbal product responsible for glucose control and upregulation of insulin signal transduction. We performed an experiment on DLBS3233 to examine the response of TNF-α and NF-κβ and the expression of GLUT 1 and GLUT2. Methods: A total of 30 adult male Wistar rats were randomly divided into six groups (n=5 per group): nondiabetic rats in the control group (group 1); untreated diabetic rats (group 2); diabetic rats treated with DLBS3233 4,5mg/kgBW (group 3); 9mg/kgBW (group 4); 18mg/kgBW (group 5), and diabetic rats treated with pioglitazone (group 6). Immunohistochemistry was performed to examine the expression of GLUT1 and GLUT2 in the pancreas and expression of TNF-α and NF-κβ in the kidney. The data was then analyzed by ANOVA. Results: In the DBLS3233 group, reduced expression of both TNF-α and NF-κβ was seen through immunohistochemistry, whereas GLUT1 and GLUT2 were intensified compared to untreated groups. From statistical analysis, we obtained significantly lower expression of TNF-α and NF-κβ, as well as enhanced GLUT1 and GLUT2 expression compared to untreated groups (p Conclusions: DBLS3233 significantly reduces the inflammatory process and enhances the expression of GLUT1 and GLUT2 diabetic rats.

Background: There is an essential need for cardiomyocyte regeneration among patients with heart failure. Transplantation of dedifferentiated fat (DFAT) cells may lead to an improvement of cardiomyocyte regeneration among heart failure patients. We believe that DFAT cells are promising candidate cell sources for cardiac regeneration. However, the pathway underlying how DFAT cells of the adipose lineage differentiate into mature cardiomyocytes isn’t fully understood. Methods: We conducted an experimental laboratory study on isolated DFAT cells from adipose tissue of healthy adults. Then, we treated cells with different concentrations of reversine (10, 20 and 40 nM), and performed RNA extraction and cDNA synthesis. Next, we used a ceiling culture method based on the buoyancy properties of mature lipid-filled adipocytes. Stemness expression (Octamer-binding transcription factor 4 [Oct4], brachyury, Fetal liver kinase 1 [Flk-1]) was quantified by reverse transcription-quantitative (RT-q)PCR, while cardiomyocyte expression (Transcription factor GATA-4 [GATA4] and cardiac troponin T [cTnT]) was quantified by immunocytochemistry. Results: ANOVA with Tukey’s post-hoc found that 10 nM reversine increased greater Flk-1 expression compared to the control group (MD: 5.037 + 0.998; p < 0.001), but there were no significant changes among Oct4 (MD: 0.013 + 1.244; p = 0.99) and brachyury expression (MD: 0.157 + 0.084; p = 0.252). Kruskal-Wallis revealed that the expression of GATA4 (1.65 [0.41-1.98] to 0.015 [0.007-0.034]; p = 0.017) reduced significantly from day 7 until day 21 and cTnT (5.07 [6.62-8.91] to 8.22 [6.81-9.40]; p = 0 .001) increased significantly from day 7 until day 21. Conclusions: Reversine could increase the expression of Flk-1, but it was unable to stimulate the expression of Oct4 and brachyury related to stem cell-ness. An optimal concentration of 10 nM reversine may have the greatest effect on enhancing the differentiation of DFAT cells into mature cardiomyocytes, as indicated by higher cTnT expression between cells.

Background: Chronic physical stress has many effects on the nervous system and can cause structural changes in different parts of the brain and hemomodulatory, including hormonal. Current pharmacotherapeutic treatments have limited efficacy and are associated with many deleterious side effects Aim: The aim of this research is to determine how Apis dorsata forest honey administration affects follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels in rats who are subjected to forced swim tests as a model of chronic physical stress placed in a container filled with water from which it cannot escape. Methods: This was an experimental laboratory study with 32 rats divided into four treatment groups: control (C), Treatment 1 (T1) with a forced swim test + honey (2 g/rat/day), Treatment 2 (T2) with a forced swim test + honey (4 g/rat/day), and Treatment 3 (T3) with a forced swim test + honey (6 g/rat/day). All treatments were administered for 14 days. Then, blood was taken for FSH and LH serum tests, and a one-way ANOVA and Duncan test were used to statistically test the data analysis. Results: The results of this study indicate that the administration of forest honey had no significant effect (p > 0.05) on the FSH parameter, but there was a significant decrease in LH levels in the T2 and T3 groups (p < 0.05). Conclusion: It can be concluded that giving forest honey to rats who were subjected to a 14-day forced swim test had no effect on FSH and LH levels. In rats given a forced swim test as a model of chronic stress, administration at doses of 4 and 6 g/rat/day reduced LH serum levels. Thus, giving forest honey could maintain reproductive health in rat that experience chronic stress. [ABSTRACT FROM AUTHOR]