14th Day Research Articles

The Impact of Fermented Milk Products Containing Bifidobacterium longum BB536 on the Gut Environment: A Randomized Double-Blind Placebo-Controlled Trial

Probiotics, particularly those native to the gut microbiota, have a profound influence on the gut environment. In this study, we conducted a randomized placebo-controlled, double-blind, parallel-group comparison trial to investigate the effects of Bifidobacterium longum BB536 (B. longum BB536) on the fecal microbiota and metabolite compositions in healthy individuals. We compared the effects of fermented milk produced solely with Streptococcus thermophiles and Lactobacillus bulgaricus (placebo group) and fermented milk supplemented with B. longum BB536 (BY group). Our findings revealed a significantly greater relative abundance of Faecalibacterium in the BY group than in the placebo group by the 3rd day, a trend that persisted until the end of the trial on the 17th day. Additionally, the BY group presented significantly increased concentrations of tryptophan (Trp), Indole-3-lactic acid, and Indole-3-aldehyde on the 17th day. A significant positive correlation was observed between the relative abundance of Faecalibacterium and the number of viable B. longum BB536 bacteria in the feces. The concentrations of Trp and Indole-3-acetic acid were also significantly correlated with the number of viable B. longum BB536 bacteria in the feces. Our results suggest that B. longum BB536 intake can modulate the gut microbiota and metabolite profiles, which are general indicators for monitoring the gut environment, potentially conferring health benefits to the host.

Impact of Probiotic and Bioprotective Cultures on the Quality and Shelf Life of Butter and Buttermilk

In this study, butter and the corresponding buttermilk samples were produced with cream fermented by aromatic (A) or probiotic (P) cultures with or without complementary bioprotective culture (BC). The samples were characterised for their composition and colour parameters. Texture and rheological properties were evaluated at 10 and 20 °C. Microbiological (lactobacilli, lactococci, and yeast and mould counts) and sensory (aroma, taste, texture, and global evaluation) analyses were also performed. All butter sample characteristics were in accordance with the Portuguese standard. Regarding colour, the sample obtained with cream fermented by probiotics plus bioprotective culture (PBC) presented higher L* and b* values, indicating a slightly higher yellow chroma. However, colour differences (ΔEab*) in the butter samples were, in most cases, not detectable by a common observer. Butter samples P and PBC presented a significantly higher viscous modulus and consequently higher dynamic viscosity values (ca. log 6.5 Pa.s at 10 °C and log 5 Pa.s at 20 °C). Butter samples presented a pseudoplastic behaviour, and rheological parameters showed a high dependence on temperature. The counts of lactobacilli and lactococci in the butter samples were of the order of log 7–8 CFU/g, while yeast and mould counts were lower than log 2 CFU/g until the 30th day of storage, after which they showed a sharp increase to ca. log 5 CFU/g between the 30th and the 60th days of storage. Regarding sensory attributes of butter, sample P received the highest overall liking, followed by samples ABC and PBC. Sample A was the least appreciated. Buttermilk samples presented significant differences regarding their composition, viscosity, and colour parameters. In all cases, lactobacilli and lactococci counts exceeded log 7 CFU/mL after 30 days of storage, but yeast and mould counts were of the order of log 5–6 CFU/mL at the 15th day of storage. Samples P and PBC presented yeast and mould counts ca. 1–2 log cycles lower than samples A and ABC, indicating the potential of probiotic and bioprotective cultures to extend the shelf life of the product. Regarding the sensory attributes of buttermilk, samples P and PBC received the highest overall liking, followed by sample ABC. Sample A received the lowest scores, as had occurred with the butter samples. However, in all cases, the scores obtained by the buttermilk samples were lower compared to the ones of the corresponding butter. It can be concluded that both probiotic butter and buttermilk present high levels of lactobacilli and lactococci and can maintain their probiotic potential throughout the storage period.

Correlations between Metabolic Parameters, Blood Cytokine Levels, and Behavioral Parameters in Male and Female Rats of Different Age after Intrauterine Stress.

We studied interrelationships between behavioral parameters, metabolic processes, and cytokine content in the blood of male and female rats at different stages of postnatal ontogeny after intrauterine stress (mothers were forced to swim in cold water from the 10th to the 16th day of pregnancy). Correlations between behavioral and metabolic parameters in prenatally stressed rats were revealed at an earlier age (day 21 of life) than in controls. In comparison with intact rats, in males exposed to intrauterine stress no relationships between these parameters were revealed at older age, while in females, their direction and character change on day 60 of life. In prenatally stressed males, correlations of the blood levels of IL-6 were revealed: inverse relationships with behavioral parameters on the days 21 and 30 and direct relationships with metabolic parameters on day 60 of life. In prenatally stressed females on day 30 of life, we observed negative correlations between IL-4 levels and metabolic parameters that were absent under normal conditions. Thus, intrauterine stress leads to reorganization of the relationships between the parameters of metabolic and immune processes, which are essential for the purposeful behavior of mammals. The effects of stress seem to depend on the sex of the offspring and stage of postnatal development.

Potential ameliorative effect of Dapagliflozin on systemic inflammation-induced cardiovascular injury via endoplasmic reticulum stress and autophagy pathway.

Dapagliflozin (DPG) is a sodium-glucose cotransporter-2 inhibitor and is used in the treatment of diabetes. In this study, we aimed to investigate the effect of DPG on cardiotoxicity caused by systemic inflammation via endoplasmic reticulum (ER) stress and autophagy. Four groups of thirty-two Wistar Albino rats were created: Control (1ml oral physiological saline for five days and intraperitoneal saline on the 5th day), LPS (1ml oral physiological saline for five days and intraperitoneal 5mg/kg of LPS on the 5th day), LPS + DPG (10mg/kg of DPG orally for five days and 5mg/kg of LPS intraperitoneally on the 5th day), and DPG (10mg/kg of DPG orally for five days and 5mg/kg of SF intraperitoneally on the 5th day). Histopathological and immunohistochemical analyses were performed on heart and aorta tissues. ER stress and autophagy gene markers in heart tissues were evaluated by RT-qPCR. Oxidative stress in heart tissues and serum cardiac enzymes were analyzed by spectrophotometric method. The heart and aortic tissues of the LPS group showed increased expressions of Tumor Necrosis Factor-α (TNF-α) and Caspase-3 (Cas-3), along with mild hyperemia, slight inflammatory cell infiltrations, and myocardial cell damage. The heart tissues also showed genetically increased expressions of include binding immunoglobulin protein (BiP/ GRP78), protein kinase RNA-like ER Kinase (PERK), inositol-requiring enzyme 1 (IRE-1), activating transcription factors 4 (ATF-4), activating transcription factors 4 (ATF6), C/EBP homologous protein (CHOP), and BECLIN 1. Furthermore, Creatine kinase-MB (CK-MB) and Lactate dehydrogenase (LDH) levels in blood tissue significantly increased, according to biochemical analysis. With DPG therapy, all of these findings were reversed. In conclusion, DPG protects against the cardiotoxic effect of systemic inflammation with its antioxidant and anti-inflammatory properties by regulating ER stress and autophagy pathways.

Models of Patient Derived Hematopoietic Stem Cell Xenografts for Assessing Individual Human Radiosensitivity

There are different approaches to assessing the human individual radiosensitivity. In this study, individual radiosensitivity was assessed in terms of survival and recovery of human hematopoietic stem cells (HSC) after acute gamma irradiation of humanized mice. Immunodeficient NOD SCID mice were transplanted with cord blood HSC intravenously, peripheral or umbilical cord blood HSC intraosseously. The estimated D0 value for human HSCs was 1.19 Gy (95٪ CI 0.90 to 1.74), 0.99 Gy (95٪ CI 0.87 to 1.15), and 0.93 Gy (95٪ CI 0.61 to 1.91) for the three methods of obtaining humanized mice, respectively. For all three methods of mouse humanization, statistically similar models that describe the dependence of HSC survival on the acute gamma irradiation dose in the range of 0.5—1.5 Gy were obtained. Thus, intraosseous administration of peripheral blood HSCs to immunodeficient mice can be effectively used to assess the response of human HSCs to radiation exposure. Comparison of the HSC number (CD34+ cells) and their descendants (CD45+ cells) in non-irradiated and irradiated mice humanized with cells from the same donor on days 3 and 14 after irradiation makes it possible to evaluate the processes of radiation-induced death and recovery of HSCs. A coefficient calculated as the ratio of the proportion of HSCs among all human cells in the bone marrow of humanized mice on the 14th day to the proportion of HSCs on the 3rd day after irradiation was proposed to assess the response to radiation exposure. This coefficient had an inverse linear dependence on the radiation dose, differed in mice with increased and normal radiosensitivity, and increased with the use of the radioprotector cysteamine in humanized mice. We propose to use this coefficient for a comparative assessment of human radiosensitivity.

Post-stroke fatigue and its correlation with family functioning in patients who have experienced a first episode of stroke

ObjectiveThis study aimed to analyse the relevant factors that may affect post-stroke fatigue (PSF) in patients with stroke and further explore the correlation between family functioning and PSF.MethodsPatients who had experienced a first episode of stroke and were admitted to the Department of Neurology of the First Affiliated Hospital of Wenzhou Medical University were rigorously screened. The general data and family functioning of the patients on admission were collected, and their family adaptation, partnership, growth, affection and resolve scores and their PSF on the 5th day of admission were collected using the fatigue severity scale (FSS). Multiple linear regression analysis was then utilized to explore the factors affecting PSF in patients with stroke.ResultsA total of 220 questionnaires were distributed, and 220 were returned, with 212 valid questionnaires and a valid return rate of 96.4%. These 212 patients had a family functioning score of 6.58 ± 2.00 and an FSS score of 36.62 ± 10.96. Spearman’s correlation analysis showed negative correlations between the FSS scores and the adaptation, partnership, growth, affection, resolve and family functioning scores (r = −0.380, −0.505, −0.470, −0.303, −0.281 and −0.712, respectively; p < 0.001). Furthermore, multiple linear regression analysis showed that family functioning (β′ = −0.516), marital status (β′ = −0.244), household income (β′ = −0.185), literacy (β′ = −0.181) and activities of daily living (β′ = −0.084) were influential factors for PSF in patients with stroke (p < 0.05).ConclusionThere is a significant negative correlation between family functioning and PSF, suggesting that better family functioning may help mitigate the severity of post-stroke fatigue. Healthcare providers should identify interventions to help patients and families address fatigue, boost disease recovery, promote patients’ physical and mental health and improve their quality of life.

Effectiveness of Contract-relax Vs Post-isometric Relaxation in Hamstring Tightness in Healthy Young Adults

Objective: The objective of the study was to compare the effectiveness of Post-isometric Relaxation Technique VS Contract-Relax Technique to improve hamstring flexibility in healthy young adults. Method: Total 30, male and female participants,15 in each group, i.e. Group A & Group B between the age group 18 to 24 years were selected from RJS Group of Institutes, Kopargaon using random sampling method according to the inclusion and exclusion criteria. Pre-assessment of active knee extension range of motion (ROM) was done using Active Knee Extension (AKE) test with the universal full- circle goniometer and Reading 1 was noted for each subject. The subjects in Group A were given Post-isometric relaxation, 8 seconds for 12 days and Group B were given Contract-Relax Technique, 17 seconds for 12 days. Reading 2 & Reading 3 were noted on the 6th and 12th day of the treatment session respectively. Results: The results indicate a statistically significant difference between Group A and Group B. The t-value of 5.908 and p-value of less than 0.01 provides significant support for the alternative hypothesis of Hypothesis 1. The Mean values of Group B [4.44] is significantly higher than that for Group A [2.36] which suggests that Contract-Relax technique showed more significant increase in the Active knee extension range of motion than the Post-isometric relaxation after 12 days of treatment protocol. Conclusion: The statistical data results concluded that Contract-relax technique is more effective than Post-isometric relaxation technique in improving the hamstring flexibility in healthy young adults.

Evaluation of the efficiency of the hepatoprotector glucuronate in toxic hepatitis of various etiology

The objective was to study the effectiveness of the use of hepatoprotector glucuronate at the early stage of intensive therapy of toxic hepatitis of various etiologies.Materials and methods. We studied 120 patients with toxic hepatitis who were treated at the Republican research center for emergency medicine in 2022–2023. The study was carried out in two groups of patients. Group I – 60 patients admitted in 2023, who, in addition to traditional therapy, received a complex hepatoprotector glucuronate (Jetepar®)*. Group II (comparison group) – 60 patients who applied in 2022 and received only traditional therapy. Biochemical blood parameters were studied upon admission and over time on day 5. The severity of intellectual impairment was assessed using cognitive scales and the Reitan test on days 2 and 5 from the start of treatment.Results. Upon admission, both groups showed signs of toxic liver damage. In dynamics by the 5th day in patients of group I, there was a decrease in ALT by 63.7%, AST by 66.4%, alkaline phosphatase by 54.2% from the initial values, which was 3.9 and 2.6, 2. 1 lower than in the comparison group. The level of free ammonia in patients of group I on the 5th day decreased by 52%, and lactate by 57% from the initial level, almost to the physiological norm, while in the comparison group (II) only by 24.8% and 38.1%, which was 2.2 and 1.5 times worse than in the main group. Screening of the level of intelligence using two cognitive scales and the Reitan test showed that in patients in the comparison group, cognitive function indicators on the 5th day were 1.4, 1.5 and 1.2 times lower than in the main group.Conclusion. The use of the hepatoprotector glucuronate improves liver parameters and cognitive functions in patients with toxic hepatitis.

Physical Symptoms and Coping Strategies With Earthquake Stress Among Adults Living in Tent Cities: A Cross-Sectional Study.

This study aimed to examine the physical symptoms, coping strategies for earthquake stress and factors predicting coping strategies among adults staying in tent cities after an earthquake. A cross-sectional design was conducted. The study sample consisted of 706 adults residing in tent cities in three cities of earthquake-affected regions. Data were collected through face-to-face surveys and the Earthquake Coping Strategies Scale, starting from the 16th day after the earthquake in February-March 2023. Independent samples t-test, one-way analysis of variance, McNemar test and multiple linear regression analysis used for data analysis. The first three physical symptoms with the highest frequency after the earthquake were insomnia, frequent waking and difficulty falling asleep, respectively. Individuals with chronic illnesses, lower education levels, no previous experience with earthquakes and having undamaged homes more frequently used religious coping strategies. Individuals with higher incomes, employed individuals, non-smokers and those with having undamaged homes more frequently used positive reappraisal strategies. The present study concluded that physical symptoms were common among individuals in the post-earthquake period and that many individuals adopted religious coping strategies. Nurses can develop effective interventions for preventing physical and mental health problems in the post-earthquake period. They can contribute significantly as caregivers, leaders, researchers and advisors in health monitoring and improvement practices for individuals after earthquakes. The results of the study are instructive for designing prevention and improvement practices to minimise future destructive effects of earthquakes. Individuals mostly experienced sleep problems and preferred different coping styles in the early post-earthquake period. The physical and mental health of individuals can be improved by early intervention and encouraging effective coping methods after an earthquake. STROBE checklist. No patient or public involvement.

POSSIBILITIES OF NON-STANDARD METHODS FOR TREATING INFECTED WOUNDS

Summary. Introduction. Some representatives of the microcosm can secrete a variety of biologically active substances that have a detrimental effect on other groups of microorganisms. The geniuses of humanity noticed this, and today medicine has about 1600 antibacterial drugs. However, the struggle for survival at all levels continues. Many bacteria have become resistant to existing antibiotics. Taking this into account, the treatment of purulent-inflammatory diseases faces insurmountable difficulties. Contaminated ulcerative processes and various wounds of the legs are accompanied by suppuration, the treatment of which is extended by weeks and months. The above justifies the relevance of the problem. Aim: to study the possibilities of using non-standard methods for treating purulent and infected wounds and ulcers. Materials and methods. In experimental conditions (patent), we studied and tested the treatment of purulent ulcers (S=2.0–42.0 cm2) and wounds (S=22.0–48.0 cm2) in 21 patients without the use of antibiotics. Apatite polymer drainage bandages (APDB) were applied under compression conditions. Results and discussion. The dressing was changed after 12–24–48 hours, depending on the exudation level. The clinical effect was observed from the 4th day. Epithelization of wound surfaces was observed on days 14–21, of ulcerative defects – on days 18–32. Conclusions. APDB based on zinc oxide may be an alternative to existing standard treatment methods for purulent diseases. Funding. This study was carried out with grant support from the National Research Foundation of Ukraine (project No. 2023.04/0109).

Differentiation of human umbilical cord mesenchymal stem cells into functional intestinal epithelial cells via conditioned medium co-culture

BackgroundThe induction of stem cell differentiation to generate intestinal epithelial cells (IECs) with absorptive functions offers significant therapeutic potential for treating conditions such as Crohn’s disease, ulcerative colitis, radiation enteritis, and other refractory intestinal epithelial injuries. Human umbilical cord mesenchymal stem cells (hUC-MSCs) are capable of differentiating into functional IEC-like cells. ObjectiveThis study aimed to induce the differentiation of hUC-MSCs into IECs using a conditioned medium co-culture method. MethodA culture medium derived from human IECs was used as the inductive medium to facilitate the differentiation of hUC-MSCs into IECs. The cellular morphology was assessed using inverted microscopy, and the expression of IEC markers, including Villin, CK20, CK8, and CK18 proteins, was analyzed via immunofluorescence staining. Furthermore, the expression levels of IEC markers, such as KRT18, were quantified using real-time quantitative PCR analysis. The functionality of the differentiated IECs in terms of sucrase secretion was assessed through sucrase activity assays. ResultsBy the 14th day of induction, hUC-MSCs exhibited a morphology similar to IECs and exhibited the expression of IEC markers, including the KRT18 gene and Villin, CK20, CK8, and CK18 proteins. Sucrase activity assays further confirmed that the differentiated cells demonstrated significant sucrase activity. ConclusionThe conditioned medium co-culture method effectively induced the differentiation of hUC-MSCs into functional IECs.

The Impact of Homogenization Techniques and Conditions on Water‐In‐Oil Emulsions for Casein Hydrolysate–Loaded Double Emulsions: A Comparative Study

ABSTRACTThis study aims to evaluate homogenization techniques and conditions for producing stable, small droplet‐size water‐in‐oil (W/O) emulsions intended for incorporation into casein hydrolysate–loaded double emulsions. Three commonly used homogenization methods; rotor–stator, ultrasonic, and high‐pressure homogenization were individually optimized utilizing response surface methodology. Instances of over‐processing were observed, particularly with the rotor–stator and ultrasonic homogenizers under specific conditions. Nevertheless, optimal conditions were identified for each technique: 530 s at 17,800 rpm agitation speed for the rotor–stator homogenizer, 139 s at 39% amplitude for the ultrasonic homogenizer, and 520 s at 1475 bar for the high‐pressure homogenizer. Subsequently, the W/O emulsions produced under optimal conditions and their respective W1/O/W2 double emulsions were compared. The rotor–stator and high‐pressure homogenized W/O emulsions exhibited comparable narrow droplet‐size distributions, as indicated by similar Span values. However, high‐pressure homogenization failed to sufficiently minimize droplet size. Ultrasonic homogenization resulted in droplets at the 1‐μm scale but yielded more polydisperse droplet‐size distribution. According to TOPSIS analysis, an emulsion with a viscosity of 93.1 cP (centiPoise), a stability index of 93.8%, a D(90) of 0.67 μm (0th day), and a D(90) of 0.75 μm (30th day) produced using a rotor–stator was selected. Additionally, double emulsions containing primary emulsions prepared with the rotor–stator method demonstrated higher viscosity, narrower droplet‐size distribution, and lower creaming compared to other samples. This investigation sheds light on the influence of homogenization techniques on emulsion properties, providing valuable insights for optimizing double emulsion formulations.

Exploring the potential therapeutic benefits of 7-methoxy coumarin for neuropathy pain: an in vivo, in vitro, and in silico approach.

7-Methoxycoumarin (7-MC) is well recognized for its anti-inflammatory and anti-nociceptive actions. Its capacity to lessen neuropathic pain hasn't been documented yet. Hence the impact of 7-MC on vincristine-induced peripheral neuropathic pain in rodents was investigated. The investigation also looked at the impact of 7-MC in reducing neuropathic pain via voltage-gated calcium channels and phospholipase enzyme inhibition using pertinent in vitro and in silico methods. Vincristine (0.1mg/kg, i.p., daily) was administered continuously for 7 days to induce peripheral neuropathic pain in mice, with cold allodynia and thermal hyperalgesia and evaluated on the 8th day using the acetone bubble test and hot water tail immersion test. In order to derive the mechanistic approach for ameliorating neuropathic pain, the role of 7-MC in the inhibition of the phospholipase enzyme, gene expression studies on voltage-gated calcium channels using mouse BV2 microglial cells and in silico studies for its calcium channel binding affinity were also performed. The test compounds reduced vincristine-induced cold allodynia and thermal hyperalgesia in mice in a dose-dependent experiments. In vitrostudies on phospholipase inhibition by 7-MC showed an IC50 of 27.08µg/ml and down-regulated the gene expression of calcium channels in the BV2 microglial cell line. In silico docking scores for 7-MCwere higher than the standard drug gabapentin. The compound 7-MC has shown promise in alleviating vincristine-induced peripheral neuropathicin mice. Studies conducted in parallel, both in silico and in vitro have demonstrated that 7-MC effectively reduces neuropathic pain. This pain reduction is achieved through two mechanisms: inhibiting the phospholipase enzyme and blocking voltage-gated calcium channels.

Pharmacokinetics of the drug Amoxyantar in pigs

The article presents the results of pharmacokinetic studies of the drug Amoxiyantar in pigs. The results obtained indicate that amoxicillin is distributed throughout the tissues of animals, providing therapeutic concentrations in the studied dosage, ensuring effective distribution in the organs and tissues of the body of pigs, and also confirm the bioavailability of succinic acid with a high concentration in the blood plasma for an average of two hours. Based on the results of determining the dynamics of excretion of residual amounts of amoxicillin, it was detected in only one animal in the kidneys at a concentration of 54.9 μg/kg and in muscle tissue at a concentration of 25.2 μg/kg. In the remaining tissue samples of pigs obtained two days after application of the drug, as well as in all tissue samples of pigs obtained from the sixth day to the 18th day after application of the drug, amoxicillin was not detected.

Lactobacillus rhamnosus Helps to Reduce the Duration of Bleeding in Breastfed Infants with Allergic Proctocolitis.

Background: Cow's milk protein allergy (CMPA) is the most common food allergy in infants. The current standard of care (SOC) involves eliminating the allergen from both the infant's and mother's diet for 2-4 weeks. The purpose of this study is to assess the effectiveness of Lactobacillus rhamnosus (Ramnoflor) in reducing the duration of bleeding in these infants. Methods: This randomized clinical trial was conducted at Bahrami Children's Hospital on breastfed infants who were diagnosed with CMPA and had a positive occult blood (OB) test. Patients were randomly assigned to either the control or case groups. All patients received SOC therapy, with the case group receiving Ramnoflor and the control group receiving a placebo. Data were recorded on the checklist, and the children were followed and visited three times during the study, with an OB assessment at each visit. Results: The study enrolled 48 infants. Among the infants in the case group, the OB test was positive in four cases (8.3%) on the fifth day. However, there were no positive cases on the 14th and 30th days. The prevalence of this test was significantly lower in patients who received probiotics compared to the control group on the fifth day (p < 0.001). There were no positive OB tests on the 14th and 30th days in any of the groups, and no significant difference was observed between the groups. Conclusion: The addition of L. rhamnosus to SOC therapy led to a decrease in the duration of rectal bleeding in infants with CMPA compared to the control group.

Innovative process for improving functional and sensory quality of black garlic

Black garlic (BG) is an aged product high in brown melanoidin pigment and functional quality, however, the long aging process limits its commercial production. The present investigation was undertaken to develop an innovative process for improving the functional quality of BG as well as shortening the aging duration. Peeled garlic was subjected to two pre-treatment steps viz. Freeze-thaw (FT) and blanching (BL) followed by ascorbic acid (AA) infusion and effects were monitored during aging. Non-enzymatic browning, the main visible indicator of MR was significantly (p < 0.05) high in FT garlic due to high infused AA content. The pre-treated garlic showed significantly (p< 0.05) high fructose content (6-7-fold) over the control on the 6th day of aging. The treatment also enhanced the total phenolic content (2.5–3.0-fold) and antioxidant activity (14-fold) while reducing the processing time from 18 to 12 days. The soluble solid content (58.7 °B) and Fast index (fluorescent of advanced Maillard reaction products and soluble tryptophan) was maximum (305.2) in FT garlic on the 12th day. FTIR confirmed a strong peak at 1515 cm−1 depicting the dark nature of the BG subjected to FT. The innovative short process saved 50% of energy in production and was 43% more economical compared to its control counterpart. Overall sensory quality of FT garlic in terms of color, flavour and texture was on par with control except BL sample which showed high stickiness and low score values. Overall results suggest that FT treatment followed by infusion of ascorbic acid accelerated the Maillard reaction (MR), and improved the functional quality of BG.

The role and mechanism of NRG1/ErbB4 in inducing the differentiation of induced pluripotent stem cells into cardiomyocytes

BackgroundWe aimed to investigate the effect and potential mechanism of enhancing Neuregulin1 (NRG1)/v-erb-b2 avian erythroblastic leukemia viral oncogene homolog 4 (ErbB4) expression on the differentiation of induced pluripotent stem cells (iPSCs) into cardiomyocytes.MethodsWe utilized CRISPR-CAS9 technology to knock in ErbB4 and obtained a single-cell clone IPSN-AAVS1-CMV-ErbB4 (iPSCs-ErbB4). Subsequently, we induced the differentiation of iPSCs into cardiomyocytes and quantified the number of beating embryoid bodies. Furthermore, quantitative real-time PCR assessed the expression of cardiomyocyte markers, including ANP (atrial natriuretic peptide), Nkx2.5 (NK2 transcription factor related locus 5), and GATA4 (GATA binding protein 4). On the 14th day of differentiation, we observed the α-MHC (α-myosin heavy chain)-positive area using immunofluorescent staining and conducted western blotting to detect the expression of cTnT (cardiac troponin) protein and PI3K/Akt signaling pathway-related proteins. Additionally, we intervened the iPSCs-ErbB4 + NRG1 group with the PI3K/Akt inhibitor LY294002 and observed alterations in the expression of cardiomyocyte differentiation-related genes.ResultsThe number of beating embryoid bodies increased after promoting the expression of NRG1/ErbB4 compared to the iPSCs control group. Cardiomyocyte markers ANP, Nkx2.5, and GATA4 significantly increased on day 14 of differentiation, and the positive area of α-MHC was three times that of the iPSCs control group. Moreover, there was a marked increase in cTnT protein expression. However, there was no significant difference in cardiomyocyte differentiation between the iPSCs-ErbB4 group and the iPSCs control group. Akt phosphorylation was significantly increased in the iPSCs-ErbB4 + NRG1 group. LY294002 significantly reversed the enhancing effect of NRG1/ErbB4 overexpression on Akt phosphorylation as well as the increase in α-MHC and cTnT expression.ConclusionsIn conclusion, promoting the expression of NRG1/ErbB4 induced the differentiation of iPSC into cardiomyocytes, possibly through modulation of the PI3K/Akt signaling pathway.

Physiological and Transcriptomic Analyses Reveal the Role of the Antioxidant System and Jasmonic Acid (JA) Signal Transduction in Mulberry (Morus alba L.) Response to Flooding Stress

In recent decades, the frequency of flooding has increased as a result of global climate change. Flooding has become one of the major abiotic stresses that seriously affect the growth and development of plants. Mulberry (Morus alba L.) is an important economic tree in China. Flooding stress is among the most severe abiotic stresses that affect the production of mulberry. However, the physiological and molecular biological mechanisms of mulberry responses to flooding stress are still unclear. In the present study, reactive oxygen species (ROS) metabolism, antioxidant mechanism, and plant hormones in mulberry associated with the response to flooding stress were investigated using physiological and transcriptomic analysis methods. The results showed significant increases in the production rate of superoxide anion (O2•−) and the content of hydrogen peroxide (H2O2) in leaves on the 5th day of flooding stress. This led to membrane lipid peroxidation and elevated malondialdehyde (MDA) levels. Antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD) exhibited enhanced activities initially, followed by fluctuations. The ascorbic acid–glutathione (AsA-GSH) cycle played a crucial role in scavenging ROS, promoting the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH). Transcriptomic analysis revealed the up-regulation of the gene-encoding antioxidant enzymes (APX, MDHAR, GPX, GR, GST) involved in ROS scavenging and stress tolerance mechanisms. Jasmonic acid (JA) levels and the expression of JA synthesis-related genes increased significantly in mulberry leaves under flooding stress. This activation of the JA signaling pathway contributed to the plant’s adaptability to flooding conditions. Proline (Pro) and soluble sugar (SS) contents increased notably in response to flooding stress. Proline helped maintain cell turgor and protected enzymes and membranes from damage, while soluble sugars supported anaerobic respiration and energy supply. However, soluble protein (SP) content decreased, suggesting inhibition of protein synthesis. The study provides insights into mulberry’s flooding tolerance mechanisms, guiding future molecular breeding efforts. This summary captures the key findings and implications of the study on mulberry’s response to flooding stress, focusing on physiological and molecular mechanisms identified in the research.

Sirt1/Nrf2/TNFα; TLR4/Myd88/NF-κB signaling pathways are involved in mediating hepatoprotective effect of bupropion in rat model of myocardial infarction

Background The aim of the current study is to identify the possible protective effect of bupropion (BUP) on liver injury in rat model of myocardial infarction (MI). BUP was administered in the presence and absence of MI. Materials and methods Thirty-two Wistar adult male rats were randomly arranged into four groups: control, BUP (30 mg/kg/day, intraperitoneal) for 28 days, isoproterenol (ISO) was injected subcutaneous (85 mg/kg) in the 26th and 27th days and BUP/ISO groups. Cardiac and hepatic enzymes were measured, also Hepatic oxidative stress indicators, as well as inflammatory and apoptotic biomarkers, were evaluated. Cardiac and hepatic histopathological examination and hepatic nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) immunohistochemical study were also detected. Results ISO significantly increased cardiac and hepatic enzymes, hepatic oxidative stress, inflammatory, apoptotic, with a histopathological picture of cardiac and hepatic damage and high hepatic NF-κB immunoexpression were detected. BUP significantly normalized the upraised oxidative, inflammatory, and apoptotic parameters, with an impressive improvement in the histopathological picture and a reduction in hepatic NF-κB immunoexpression. Conclusion BUP protects against liver injury on top of MI in rat model via modulation of Sirtuin type 1 (Sirt1)/Nuclear factor (erythroid-derived 2)-like 2 (Nrf2)/tumor necrosis factor α (TNFα); Toll-like receptor 4 (TLR4)/Hepatic myeloid differentiation primary response 88(Myd88)/NF-κB signaling pathways.

Hepatocellular Changes on Paracetamol Induced Liver Damage in Long Evans Male Rats upon Green Tea Administration

Background: The liver is a vital organ that serves a number of purposes in our body. It may be harmed by pollutants, substances with toxic effects, and long-term, unchecked drug use. Green tea is a well-liked beverage that has gained popularity recently and may have hepatoprotective properties. Objective: To observe the effect of green tea (Camellia sinensis) on paracetamol induced liver damage in Long Evans male rats. Methods: From July 1st, 2018, to June 30th, 2019, this study was conducted in the Department of Physiology at Sir Salimullah Medical College (SSMC), Dhaka. For the purpose of the study, thirty (30) male Long Evans rats, weighing between 150 and 200 grams and 90 to 120 days old, appeared to be in good health. They were split into two groups after 14 days of acclimatization: Group A, which served as the control group, and Group B, which served as the experimental group (green tea pretreatment and paracetamol treated). Group A1 (baseline control group) and group A2 (paracetamol treated control group) comprised the subdivided control group. There were ten rats in each of these groups. For 28 days, the rats were all fed a baseline diet. The baseline control group was given normal saline (20 ml/kg/day) orally every day for 28 days in addition to their basal diet. For the final three days of the study—the 26th and 28th—the paracetamol-treated control group was given oral paracetamol at a dose of 1.5 g/kg/day. During the final three days of the study period (the 26th and 28th days), the experimental group was given oral paracetamol (1.5g/kg/day) and an ethanolic extract of green tea (500 mg/kg/day) for a total of 28 days. On day 29, every rat was sacrificed. Following the sacrifice, liver samples were taken. Hepatic contents of malondialdehyde (MDA) were measured by using standard laboratory method. Histopathology of liver was also done by using standard laboratory procedure in the department of pathology, SSMC. Version 22 of the Statistical Package of Social Science (SPSS) for Windows was used to do the statistical analysis. The data were displayed as mean±SD. To compare the results, one-way ANOVA, post hoc Bonferroni, and Chi-square tests were used, where appropriate. A p value of less than 0.05 was regarded as significant. Result: When compared to the baseline control group, the mean malondialdehyde concentration in the liver was significantly (p) greater in the green tea pretreatment, paracetamol treated, and control groups who received paracetamol treatment. Once more, the groups treated with green tea and paracetamol had mean malondialdehyde concentrations in their livers that were significantly (p) lower than those of the paracetamol group. Additionally, 0% of the rats in the baseline control group, 100% of the rats in the paracetamol-treated group, and 30% of the rats in the green tea-pretreated and paracetamol-treated group had abnormal histological findings of the liver. Conclusion: This study revealed that green tea has hepatoprotective effect against paracetamol induced liver damage in Long Evans male rats. Bangladesh Crit Care J September 2024; 12 (2): 145-152