Adult Rats Research Articles

Antidiabetic, Antioxidant, Antinociceptive, and Anti‐Inflammatory Potential of Chemically Characterized Extract of Limeum Indicum Stocks ex T. Anderson (Aizoaceae). In Vitro, In Vivo, and In Silico Investigations

AbstractLimeum indicum (LI), a plant from the Aizoaceae family, has garnered interest due to its potential health benefits. In this study, we explore the antidiabetic, antioxidant, antinociceptive, and anti‐inflammatory properties of a chemically characterized whole extract from LI. In vitro assays included DPPH and reducing power tests to assess antioxidant activity. For in vivo investigations, adult Wistar rats were used. Alloxan‐induced diabetic rats received oral doses of LI whole extract (ethanol (LI‐EE), DCM (LI‐DE), and n‐Hexane (LI‐HE)) or glimepiride for 21 days. Carrageenan‐induced inflammation and nociceptive effects were also evaluated. Molecular docking studies aided in identifying potential targets for the extract's bioactivity. Phytochemical analysis revealed the presence of cardiac glycosides, flavonoids, phenolics, saponins, and fixed oils in the extract. Both DPPH and reducing power assays demonstrated substantial total phenolic and flavonoid content, indicative of strong antioxidant activity. The extracts dose‐dependently reduced blood glucose levels in alloxan‐induced diabetic rats and mitigated inflammation and nociception. Furthermore, the significant antidiabetic and anti‐inflammatory activities of n‐hexane extract, suggested to identify the potential components by employing molecular docking studies.

Tempol mitigates inflammation, oxidative stress, and histopathological alterations of cadmium-induced parotid gland injury in rats

Cadmium (Cd) is a potent environmental pollutant that causes functional and structural damage to the salivary glands. Tempol (TEM) has powerful antioxidant activity that can potentially preserve organ function. Aims: This study was designed to investigate the protective effects of TEM on Cd-induced toxicity in rat parotid salivary glands. Materials and methods: Twenty-four adult Wistar male rats were randomly assigned to four equal groups: control, TEM (27.5 g/100 ml), Cd (0.6 g/100 ml), and TEM plus Cd (at the same doses). All treatments were dissolved in distilled water and administered subcutaneously four times a week for four weeks. Parotid gland tissues were isolated and subjected to molecular and histo-biochemical assessments. Key findings: TEM exerted a prophylactic effect against Cd-induced toxicity in the parotid glands by controlling inflammation through the downregulation of toll-like receptor 4/myeloid differentiation primary response 88/nuclear factor kappa B/ interleukin-1 beta mRNA expression, upregulation of aquaporin-5 mRNA expression, improvement of the oxidant/antioxidant status in the parotid gland, mitigation of endoplasmic reticulum stress, and repair of the associated histological and ultrastructural abnormalities. Significance: TEM protects against Cd-induced toxicity in the parotid glands of rats, attributable at least in part to its anti-inflammatory and antioxidant properties, as well as its ability to inhibit ER stress and facilitate glandular repair. However, the protective effects of TEM did not reach the levels observed in the control group. TEM could be a promising clinical candidate for protecting the salivary glands, particularly in high-risk groups such as workers exposed to Cd and cigarette smokers.

Functional Adaptation in the Brain Habenulo-Mesencephalic Pathway During Cannabinoid Withdrawal.

The mesolimbic reward system originating from dopamine neurons in the ventral tegmental area (VTA) of the midbrain shows a profound reduction in function during cannabinoid withdrawal. This condition may underlie aversive states that lead to compulsive drug seeking and relapse. The lateral habenula (LHb) exerts negative control over the VTA via the GABA rostromedial tegmental nucleus (RMTg), representing a potential convergence point for drug-induced opponent processes. We hypothesized that the LHb-RMTg pathway might be causally involved in the hypodopaminergic state during cannabinoid withdrawal. To induce Δ9-tetrahydrocannabinol (THC) dependence, adult male Sprague-Dawley rats were treated with THC (15 mg/kg, i.p.) twice daily for 6.5-7 days. Administration of the cannabinoid antagonist rimonabant (5 mg/kg, i.p.) precipitated a robust behavioral withdrawal syndrome, while abrupt THC suspension caused milder signs of abstinence. Extracellular single unit recordings confirmed a marked decrease in the discharge frequency and burst firing of VTA dopamine neurons during THC withdrawal. The duration of RMTg-evoked inhibition was longer in THC withdrawn rats. Additionally, the spontaneous activity of RMTg neurons and of LHb neurons was strongly depressed during cannabinoid withdrawal. These findings support the hypothesis that functional changes in the habenulo-mesencephalic circuit are implicated in the mechanisms underlying substance use disorders.

Rehabilitation of N, N'-methylenebisacrylamide-induced DNA destruction in the testis of adult rats by adipose-derived mesenchymal stem cells and conditional medium

Rehabilitation of N, N'-methylenebisacrylamide-induced DNA destruction in the testis of adult rats by adipose-derived mesenchymal stem cells and conditional medium

Effects of Tranexamic Acid on Human Osteoblasts as Proxy for Fracture Healing.

To investigate the effect of tranexamic acid (TXA) through in vitro culture of primary human osteoblasts (HOB) and in vivo using an operative rat femur fracture model. It was hypothesized that there would not be any effect on fracture healing in both studies. Primary HOBs were exposed to varying concentrations of TXA over different time periods. Cells were assessed for viability, metabolism, and mineralization. For the in vivo model, fractures were created in the femora of adult rats, exposed to either TXA or saline, and then assessed for healing at different time points. A modified radiographic union score for tibia was used to evaluate radiographs, callus mineralization was assessed with microcomputed tomography, and biomechanical tests were performed. Overall, HOB viability and metabolism decreased as TXA concentration and exposure time increased. However, at concentrations below 56.44 mg/mL, HOB viability was not affected. Similarly, mineralization also decreased as TXA concentration and exposure time increased. In both groups, in vivo results demonstrated increasing radiographic healing, callus mineralization, and biomechanical strength as a function of time. There was a trend for increased healing in the TXA group at 6 weeks after fracture; however, the difference compared with untreated animals was not statistically significant. Although a degradation of HOB viability and metabolism occurred with increased TXA concentrations and exposure times, clinically relevant concentrations do not adversely affect HOB viability, metabolism, or mineralization. In addition, there were no noticeable adverse effects of TXA administration in the in vivo model.

Effects of Variable Doses of Neurokinin B on Hematological and Coagulation Parameters in Adult Male Rats

Objective: To evaluate the dose-dependent effects of Neurokinin B on hematological and coagulation parameters in adult male Sprague Dawley rats. Study Design: Laboratory-based experimental study Place and Duration of Study: Gomal University and KMU Institute of Medical Sciences from February 2019 to April 2021. Methodology: Adult male Sprague Dawley rats (n=5 per group) were administered intraperitoneally with Neurokinin B at doses of 1 µg, 1 ηg and 1 ρg for 12 days consecutively. Control rats received distilled water injections. After 12 days, rats were sacrificed, and blood samples were collected from the left ventricle for hematological and coagulation analysis. Results: Bleeding time was significantly prolonged in NKB-µg (114±17.10sec) and Neurokinin B -ηg (72±22.25sec) compared to control (33±12.55sec). Clotting time (CT) was also increased (NKB-µg: 111±17.10sec; NKB-ηg: 69±17.10sec, control: 42±12.55sec), as did prothrombin time (PT) [NKB-µg: 93.60±10.92sec; NKB-ηg: 63.40±12.66sec, control: 15±4.12sec] and activated partial thromboplastin time (APTT) [NKB-µg: 94.80±14.04sec; NKB-ηg:68.60±13.74sec, control: 19.40±2.97sec]. Additionally, mean platelet volume [NKB-µg:8.94±0.36fl, control: 8.04±0.48fl] and leukocyte count (×103/µl) [NKB-µg: 12.14±0.69, control: 8.87±1.32] were significantly increased in NKB-treated animals compared to control. There was a significant increase in international normalized ratio in NKB-µg (6.86±0.80) and NKB-ηg (4.65±0.93) compared to control group (1.10±0.30). Conversely, platelet count (×103/µl) was decreased significantly in NKB-µg group (787.80±48.51) compared to the control group (960.60±58.71). Conclusion: NKB administration led to leukocytosis, thromboctopenia, and affected coagulation pathways with prolonged BT, CT, PT, APTT and INR dose-dependently.

Hepatoprotective Effect of L-Carnitine is Achieved via Activating Nrf2 and Targeting TLR4 Signaling Pathways in Thioacetamide – Induced Liver Fibrosis in Rats

Background: Liver fibrosis is a critical health problem that can result in serious illness and death. L-carnitine (LC) is a naturally occurring compound which transports fatty acids through the inner mitochondrial membrane for consequent beta-oxidation. It acts as an antioxidant to lessen cellular oxidative stress. Carnitine is essential for the transfer of long-chain fatty acids across the inner mitochondrial membrane for subsequent β-oxidation. This study was carried out to investigate the hepatoprotective effects of LC via modulation of Nrf2 signaling and TLR4 targeting pathways in rats with liver fibrosis induced by Thioacetamide (TAA). Methods: Twenty-four adult male Wister rats were assigned into four groups as follows: Group 1 served as a normal non- treated control. Rats in group 2 were injected intraperitoneally (IP) with Thioacetamide (TAA) twice a week at a dose of 200 mg/kg B.wt for 6 weeks to produce liver fibrosis. Two weeks following TAA injections, 50 and 100 mg/kg of LC were administered to the rats in groups 3 and 4, respectively concurrently with TAA injections until end of the experiment (6 weeks). Results: Intraperitoneal injection of LC decreased the levels of liver enzymes aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in rats with liver fibrosis induced by TAA. Malondialdehyde (MDA), tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), and toll-like receptor 4 (TLR4) levels were significantly decreased in LC treated groups. LC injection increased albumin, superoxide dismutase (SOD), heme oxygenase-1 (HO-1), nuclear factor erythroid 2-related factor 2 (Nrf2), and glutathione (GSH) levels. Additionally, expression of Phosphoinositide 3-kinase (PI3K) was increased and expression of TLR4 was decreased in LC treated groups according to PCR data. The biochemical findings were supported by histopathological findings. Regarding immunohistopathological examination, the LC treated groups reduced hepatic expression of caspase-3 an

Perfusate Biomarkers of DCD Cardiac Graft Quality Identified With Proteomics: Studies in an Isolated Rat Heart Model.

Heart transplantation with donation after circulatory death (DCD) enhances cardiac graft availability, but exposes hearts to potentially damaging conditions, such as warm ischemia. Normothermic machine perfusion (NMP), used for graft transportation, allows biomarker determination in perfusate. Using our isolated, rat heart model of DCD, we evaluated potent. Isolated, perfused adult male Wistar rat hearts (n = 5/group) underwent different warm ischemic durations to simulate DCD, followed by reperfusion to simulate NMP. Perfusate samples were collected after 10 min reperfusion, and proteins were analyzed using mass spectrometry. Cardiac recovery was evaluated after 60 min reperfusion. The relationship between perfusate proteins and cardiac recovery was investigated. Cardiac recovery decreased with increasing ischemic duration. Principal component analysis of perfusate proteins demonstrated segregation by ischemic group. Several proteins demonstrated an On-Off pattern, and correlated with key outcome measurements. Other proteins were released by all hearts and were confirmed as predictors of cardiac recovery, for example, heat shock protein 70 and valosin-containing protein (area under the curve [AUC] = 0.962-0.968, respectively; P < 0.05 for all). Additionally, proteins such as glycogen phosphorylase, muscle associated (AUC = 0.9632; P < 0.05) showed potential as novel biomarkers for evaluating cardiac graft quality, unlike lactate release after 10 min of reperfusion (AUC = 0.60). Multiple perfusate proteins, such as heat shock protein 70, valosin-containing protein, or glycogen phosphorylase, muscle associated, released during early reperfusion are promising as biomarkers for assessing graft quality during NMP. Perfusate proteins, as biomarkers, offer the possibility of both rapid immune detection and out-of-hospital implementation, and may provide valuable information about graft quality, especially when profiled with serial sampling during NMP.

Effect of Raha syrup on the motivational effects of morphine and CSF serotonin levels in rats

Opiates may influence motivational processes by a dysregulation of brain reward pathways that play a role in context-induced reinstatement of drug seeking. Raha syrup as herbal bioactive compounds with anti-anxiety/depressant effects may be prevented morphine-induced reward and motivation effects. Therefore, the aim of this study was to investigate the effect of Raha syrup on the rewarding effects of morphine and motivational aspects of morphine-seeking behavior in morphine-induced conditioned place preference (CPP) model and also the cerebrospinal fluid (CSF) serotonin levels following the extinction and reinstatement in rats. In this study, adult male Wistar rats received Raha syrup via oral gavage to assess of the acquisition, extinction and reinstatement of the morphine-induced CPP and as well as measurement of the CSF serotonin levels. Administration of Raha syrup during the conditioning period significantly attenuated the acquisition of morphine-induced CPP, and increased the CSF serotonin level, 6 days after cessation of treatment during extinction, while did not affect the duration of the extinction period. Also, Raha syrup during the extinction period did not affect the reinstatement of morphine-induced CPP and the CSF serotonin levels in rats. We conclude that Raha syrup attenuated the acquisition of morphine reward probably through an increase in CSF serotonin, while could not maintain the extinguished CPP and had no effect on the reinstatement. Therefore, Raha syrup as a useful adjunctive therapeutic strategy may exert a protective effect against opiate-related rewards, while it may be ineffective to prevention of motivation or drug-seeking reinstatement (relapse).

Nutritional Assessment and Sub-acute Toxicity Study in Wistar Rats Fed with a Compounded Diet of Roasted Plantain and Fish Sold in Otuoke, Nigeria

The widespread consumption of roasted plantain and fish (Bolle), a common delicacy in Bayelsa State has raised concerns over potential health risks associated with food processing contaminants such as acrylamides, polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs). The roasted plantain and fish are usually prepared by placing the peeled plantain and the dressed fish on a wired gauze and placed over a burning charcoal. The subacute toxicity study of this diet was investigated using twenty adult male rats averagely weighing 117.22 \(\pm\) 4.48 g. The rats were divided into four groups of five rats per group. Group A (Control), was fed with the standard rat chow, Group B; was fed with only the roasted plantain, Group C; was fed with only the roasted fish, and Group D; was fed with a combination of the roasted plantain and fish. The rats were given free access to the feed and portable water ad-libitum throughout the experimental protocol. Body/organ weight data were recorded before and after the completion of the experimental protocol. On day 15th of the experimental protocol, the rats were euthanized and blood specimens were obtained by cardiac puncture for biochemical analysis. The liver, kidney and heart were dissevered instantaneously and weighed. Findings from the study showed that the roasted plantain-fed group had significantly reduced body weight gain, and increased liver and kidney to body weight ratio (p&gt;0.05). The plasma enzyme activity (AST, ALT and ALP) of the plantain-fed group were also significantly increased (p&lt;0.05) while other groups showed non-significant differences (p&gt;0.05) with the control group. The antioxidant status (CAT, SOD and GPx), haematological parameters (RBC, PCV, Hb, MCV, MCH, MCHC and platelet) of the plantain-fed groups were also significantly decreased (p&lt;0.05) while other groups had non-significant changes (p&gt;0.05) when compared to the control group. Malondialdehyde levels in the roasted plantain-fed group were also significantly increased (p&lt;0.05).

STEMIN and YAP5SA, the future of heart repair?

This review outlines some of the many approaches taken over a decade or more to repair damaged hearts. We showcase the recent breakthroughs in organ regeneration elicited by reprogramming factors OCT3/4, SOX2, KLF4, and C-MYC (OKSM). Transient OKSM transgene expression rejuvenated senescent organs in mice. OKSM transgenes also caused murine heart cell regeneration. A triplet alanine mutation of the N-terminus of Serum Response Factor's MADS box SRF153(A3), termed STEMIN, and the YAP mutant, YAP5SA synergized and activated OKSM and NANOG in adult rat cardiac myocytes; thus, causing rapid nuclear proliferation and blocked myocyte differentiation. In addition, ATAC seq showed induced expression of growth factor genes FGFs, BMPs, Notchs, IGFs, JAK, STATs and non-canonical Wnts. Injected STEMIN and YAP5SA synthetic modifying mRNA (mmRNA) into infarcted adult mouse hearts, brought damaged hearts back to near normal contractility without severe fibrosis. Thus, STEMIN and YAP5SA mmRNA may exert additional regenerative potential than OKSM alone for treating heart diseases.

Determination of Astrocyte Reaction Using Glial Fibrillary Acidic Protein (GFAP) Following Aluminium Chloride Induced Hippocampus Damage of Adult Wistar Male Rats Treated with Ethanolic Extracts of Carpolobia lutea at Different Doses

Background: Aluminium is a well-established neurotoxicant involved in the etiology of neurodegenerative diseases. Carpolobia lutea, commonly called cattle stick or poor man’s candle belongs to the plant family polygalaceae. It is a small tree native to West and Central tropical Africa and it is one of the medicinal plants that play a major role in the health care system of developing countries such as Nigeria. Aim of the Study: This study was aimed at investigating astrocyte reaction in aluminium chloride induced neurotoxicity in the hippocampus in adult male wistar rats treated with ethanolic extracts of C.lutea at different doses. Materials and Methods: Thirty wistar rats weighing 180-200 g were used for this study. The animals were randomized into five groups of six rats each. Group A rats received only animal feed. Groups B,C,D and E were given 100 mg/kg bw of aluminium chloride intraperitoneally five times a week for three weeks to induce neurotoxicity.In order to reverse the neurotoxicity,. Group C was treated with 10 mg/kg bw of donepezil as standard drug while group D and E were treated with 200 mg/kg and 400 mg/kg of Carpolobia lutea respectively for 14 days. Histopathological study was done on the hippocampus of the rat brain and thereafter hematoxylin and eosin staining were carried out. Immunohistochemical studies for GFAP was done using NovocasraTM Novolink TMpolymer detection system and appropriate primary monoclonal antibodies. Image J cell counter tool was used to note the number of GFAP positive cells across all groups. Results: The result from our histopatholological study shows that lower dose (200 mg/kg/bw) of ethanolic extract of the leaf of Carpolobia lutea have better protection on the cytoarchitecture of the CA1, CA3, and dentate gyrus of the hippocampus than 400 mg/kg/bw of the plant extract. From our study, it is evident that 200 mg/kg of Carpolobia lutea reduced the GFAP immunoreactivity in the hippocampus of AlCl3 induced neurotoxicity. When we compared the result of 200 mg/kg/bw of C. lutea with 10 mg/kg per body weight of donepezil (standard drug), it was observed that C. lutea had a slight reduced GFAP immunoreactivity than the standard drug(donepezil). The lower dose, 200 mg/kg of ethanolic extract of the leaf of Carpolobia lutea have a better neuroprotective benefit than the standard drug on the hippocampus Conclusion: In conlusion, 200 mg/kg of Carpolobia lutea have the tendencies of protecting the neurons in the hippocampus from degenerating as a result of aluminium chloride induced neurotoxicity compared to the standard drug (10 mg/kg donepezil).

Study of acute and subacute toxicities and genotoxic and mutagenic potentials of the lyophilized extract of Campomanesia sessiliflora (O.Berg) mattos leaves in wistar rats

Campomanesia sessiliflora (O.Berg) Mattos is a Brazilian native plant species used in a popular medicinal tea for treating gastrointestinal, urinary, and dermatological pathologies. This study evaluated the toxicity of Campomanesia sessiliflora (O.Berg) Mattos via acute and subacute toxicity tests. It also analyzed mutagenic and genotoxic potentials by the micronucleus test, which detects genetic material damage indicating mutagenicity, and the comet assay, which assesses DNA damage levels as a genotoxicity indicator. The plant extract initially originated from the ultrasonic maceration of Campomanesia sessiliflora (O.Berg) Mattos leaves in a hydroethanolic solution. The involved animals were adult Wistar rats. Ten females were available to evaluate acute toxicity and estimate the LD50, receiving a dose of 2000 mg/kg. The subacute toxicity evaluation used 35 females and 35 males divided into seven groups: negative control (saline control – SC), positive control (cyclophosphamide control – CC), 125 mg/kg (125), 250 mg/kg (250), 500 mg/kg (500), 1000 mg/kg (1000), and the satellite group (ST). Genotoxicity and mutagenicity experiments applied bone marrow micronucleus and comet assays. Acute and subacute toxicity tests did not present behavioral, physical, and physiological changes (p≥0.05). Administering the Campomanesia sessiliflora (O.Berg) Mattos extract reduced spleen size in male and female animals, without histopathological changes. However, doses above 500 mg/kg showed significant genotoxic and mutagenic effects in the comet and micronucleus assays compared to the control group. The extract did not exhibit acute or subacute toxicity, but doses higher than 500 mg/kg indicated some level of genotoxicity and mutagenicity.

The deleterious effect of orally administered aqueous extract of &lt;i&gt;Acalypha wilkesiana&lt;/i&gt; (Red Acalypha) on female reproductive organs in Sprague Dawley rats

Acalypha wilkesiana is a herbaceous plant that has been traditionally used to treat and/or manage a variety of illnesses, including respiratory issues, diabetes, jaundice, hypertension, fever, diarrhea, and inflammation of the liver. The effects of taking Acalypha wilkesiana orally on the uterus and ovaries were examined in this study. Twenty (20) adult female Sprague-Dawley rats (165±20g) were purchased and given a two-week period to get used to the laboratory environment. Aqueous extract of Acalypha wilkesiana was given orally for four weeks at concentrations of 0, 100, 200, and 400 mg/kg of BW following acclimatization. Groups A-D provided as the control, low, medium, and high doses, respectively. The rats' weight gain and behavioral patterns were periodically examined. Following their euthanasia, blood was drawn via the ocular sinuses to measure hormonal parameters, and the uterus and ovaries were exercised to examine histology and oxidative stress markers. Significantly lower levels of FSH, LH, estrogen, and progesterone were observed in the treatment groups, while significantly lower levels of SOD and CAT and higher levels of MDA were observed. In both the 40 and 80 mg/kg groups, histological sections of the ovaries and uterus revealed vascular congestion in the ovaries and moderate to severe damage within the uteri. The findings demonstrated that long-term use of Acalypha wilkesiana increased levels of oxidative stress, altered hormone levels, and negatively affected the histoarchitecture of the uterus and ovaries.

Effect of Ketogenic Diet on Colonic Changes of Stress Induced Irritable Bowel Syndrome of Adult Male Albino Rat Model (Histological and Morphometric study)

Effect of Ketogenic Diet on Colonic Changes of Stress Induced Irritable Bowel Syndrome of Adult Male Albino Rat Model (Histological and Morphometric study)

Myelination deficiency in rats with genetically determined dopamine metabolism disorder

BACKGROUND: Attention deficit hyperactivity disorder is one of the most common neuropsychiatric disorders affecting children. The exact causes of attention deficit hyperactivity disorder are still not fully understood. Myelination deficiency in nerve fibers could be one of the possible factors in the pathogenesis of attention deficit hyperactivity disorder. Dopamine transporter knockout (DAT-KO) rats provide a good translational model for attention deficit hyperactivity disorder because they mimic behavioral symptoms of the disease: hyperactivity, cognitive impairment and compulsive behavior. This paper aims to determine abnormalities in the myelination process in the spinal cord of DAT-KO rats. Our findings can lead to a better understanding of the etiology and pathogenesis of attention deficit hyperactivity disorder. AIM: To determine the mRNA expression levels of myelination-related genes in the spinal cord of dopamine transporter knockout (DAT-KO) rats during their natural development. MATERIALS AND METHODS: The study was performed on 72 rat pups (DAT-KO, DAT-HET, DAT-WT) from 12 litters. Rat pups were born from the pairs of intercrossed DAT-HET x DAT-HET adult rats. Pups were decapitated on 7th, 14th and 21st day of their postnatal development. After that, mRNA levels of the main myelination-related genes were measured in the cervical and lumbar segments of the spinal cord using real-time PCR. RESULTS: The mRNA levels of the mag, olig2 and plp1 genes were significantly different in the cervical and lumbar spinal cords of DAT-KO rats compared to DAT-WT animals. A significant increase in the level of mag gene mRNA in DAT-HET and DAT-KO animals was observed on 14th and 21st days of postnatal development. Also, in these animals, the olig2 mRNA levels were increased on the 21st day of postnatal development. The plp1 mRNA level was increased on the 14th day, and reduced on the 21st day of postnatal development in DAT-KO animals. CONCLUSIONS: Myelination deficiency in nerve fibers in the spinal cord of DAT-KO rats is one of the effects of gene knockout. It is also of the possible causes of behavioral changes — hyperactivity, cognitive impairment and compulsive behavior.

Dietary soy protein reverses obesity-induced liver steatosis and alters fecal microbial composition independent of isoflavone level.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a major public health concern that is exacerbated by the obesity pandemic. Dietary interventions have the potential to alleviate obesity-associated MASLD through variable mechanisms, including optimizing the gut microbiota. Previously, we reported that soy protein concentrate (SPC) with low or high levels of isoflavone (LIF or HIF) protected young obese Zucker rats from developing liver steatosis. The current study was designed to test whether SPC-LIF and SPC-HIF diets would reverse liver steatosis and alter fecal microbial composition in adult obese Zucker rats with existing steatosis. Six-week-old male obese Zucker rats (n = 26) were fed a casein control diet (CAS) for 8 weeks and 7 rats were randomly selected and sacrificed to confirm liver steatosis. The remaining rats were randomly assigned to receive CAS, SPC-LIF, or SPC-HIF diet (n = 6-7/group) for an additional 10 weeks. Compared to CAS diet, feeding SPC-LIF and SPC-HIF diets resulted in significantly lower liver weight, liver steatosis score, and liver microvesicular score (p < 0.05), but did not lead to difference in body weight, liver macrovesicular score, serum ALT, or serum AST. Isoflavone levels (e.g., LIF vs. HIF) did not affect any of these measurements except in the SPC-HIF group, which had an additional decrease in liver weight (p < 0.05) compared to the SPC-LIF group. The SPC-HIF group also had significantly higher levels of the aglycone forms of daidzein, genistein, and equol as well as the total levels of daidzein, genistein, and equol compared to SPC-LIF or CAS diet fed rats (p < 0.05). The distribution of microbial communities based on measures of beta diversity of both SPC-LIF and SPC-HIF groups were significantly different to that of the CAS group (p ≤ 0.005). Alpha-diversity did not differ between any of the groups. Taken together, dietary soy protein can reverse liver steatosis in adult Zucker rats, and the reversal of steatosis is accompanied by alterations in gut microbial composition.

The STRIPAK associated kinase MST4 is a novel mediator in heart failure pathogenesis

Abstract Heart failure still poses substantial challenges in understanding its underlying molecular pathways. We recently found the mammalian STE20-like kinase 4 (MST4) significantly upregulated in human failing hearts (5.7-fold in DCM and 3.8-fold ICM) as well as in heart failure animal models (i.e. 2.5-fold in Calsarcin-1-KO, 2.5-fold in MLP-KO). MST4 is a member of striatin interacting phosphatases and kinases complexes (STRIPAK), an emerging group of multiprotein complexes responsible for integrating cellular signalling pathways. We provide the first evidence of functional STRIPAK complexes containing MST4 by co-immunoprecipitation and interactomics experiments in cardiomyocytes. Moreover, we found MST4 to interact with several intercalated disc proteins like beta-Catenin or Desmin. Adenoviral overexpression of MST4 in neonatal rat ventricular cardiomyocytes (NRVCM) results in a significant increase in cell size (+13%). This was not accompanied by the activation of fetal genes such as NPPA, NPPB or RCAN1.4, but rather by an increase in phosphorylation of Protein kinase B (PKB/Akt, 3.3-fold). Akt is usually associated with physiological hypertrophy, indicating that MST4 may be involved in protective pathways in cardiomyocytes. In support of this notion, MST4 overexpression reduced apoptotic activity (cleavage of Caspase 3 -69%, Caspase 7 -80%, Parp1 -27%) and improved both contractility (fractional shortening cell +23%, fractional shortening sarcomere +30%, time to max. contraction cell -10%, time to max. contraction sarcomere -12%) and relaxation parameters (time to max. relaxation velocity cell -12%) in isolated adult rat ventricular cardiomyocytes. In order to identify cardiac targets of MST4, we performed a comprehensive analysis of the phosphoproteome of NRVCM overexpressing MST4 compared with control-virus +/- pharmacological inhibitor at two time points. We identified more than 70,000 peptides including more than 20,000 phosphopeptides in more than 4,000 protein groups. Using linear modelling with a reduced data set containing regulated features selected by thresholds for p-value and fold change, we observed a consistent MST4 effect and a number of enriched gene ontology terms. Here, we found several potential MST4 targets and effects on protein expression levels associated with intercalated disc proteins. In human heart samples, MST4 was also localized to the intercalated disc region by immunostaining. Interesting proteins in that context that might help explain some of the observed cardioprotective MST4 effects are Connexin 43, Desmin, Ryanodine receptor, Phospholemman, Junctophilin2, Myozap or Frizzled 1. Taken together, our data imply that MST4 upregulation in human heart failure serves an adaptive role which may offer translational opportunities.Graphical Abstract

Remodeling of adult rat heart subjected to pressure overload from the neonatal period is dependent on the sex

Abstract Background Congenital heart diseases (CHD) represents the most common birth defect. Despite progress in their treatment improving acute survival, there is a paucity of data concerning the long-term consequences of CHD in adults. Purpose Here, we investigate the remodeling in adult rat hearts subjected to pressure overload from the early postnatal period. Methods We used the neonatal abdominal aortic constriction (AAC) model to analyze morphological, functional (by echocardiography), and electrophysiological long-term outcomes. Given the known differences in cardiac pathologies between males (M) and females (F), we focused on the role of sex in cardiac remodeling. Results Our data showed that both sexes exhibited a similar left ventricular weight/body weight ratio at the age of 12 weeks, indicating comparable left ventricle (LV) enlargement as a response to AAC (increase by 103 ± 9 % in M AAC and 99 ± 10 % in F AAC vs. sham-operated animals). However, the right ventricular weight/body weight and lung/ body weight ratios, indicative of LV failing, differed significantly between sexes with better values in F AAC. The favorable cardiac remodeling in F AAC was corroborated by significantly smaller LV systolic and diastolic diameters, along with a lower amount of myocardial fibrosis. Using optical mapping, we revealed preserved longitudinal and transversal conduction velocities (CVL; CVT) in females (CVL 104 ± 8 vs. 109 ± 8 cm/s and CVT 66 ± 7 vs. 59 ± 6 cm/s in F AAC and F Sham, respectively). However, compared to M Sham, the conduction velocities in M AAC showed a significant decrease (CVL 80 ± 3 vs. 92 ± 3 cm/s and CVT 35 ± 2 vs. 54 ± 3 cm/s). Similarly, the amount of connexion 43 (Cx43) and its phosphorylated state (Cx43Ser368) was preserved in F AAC while exhibiting a decrease in M AAC compared to the sham-operated controls. Thus, electrophysiological findings implicate that while the adult M AAC underwent pro-arrhythmogenic remodeling due to the elevated workload, the adult F AAC preserved physiological conduction properties. Conclusion Our data showed different consequences of the increased pressure load exposed from early after birth to the adult M and F heart. These findings highlighted the involvement of both sexes in cardiac studies.

Impact of Vitamin C and Hydroxyurea on the Reproductive System Health in Male Rats

Hydroxyurea (HU) is a medication used to treat various diseases and is also being studied for its effects on spermatogenesis. Vitamin C (VC), an antioxidant, has been shown to protect sperm cells from oxidative stress, potentially improving fertility and sperm quality. In a study involving twenty-four adult male albino rats divided into four groups, Group 1 served as the control, Group 2 received 5 mg of vitamin C, Group 3 received 100 mg of hydroxyurea per body weight, and Group 4 received a combination of vitamin C and hydroxyurea to assess essential functions of the reproductive system, including hormone levels, antioxidant markers, oxidative stress indicators, histopathology, and identification of DNA damage. The study found that hydroxyurea significantly reduced testicular weight, SOD, CAT, and GSH while increasing FSH and MDA levels and causing abnormalities in sperm morphology. Hydroxyurea also caused apparent alterations in the histological structure of the testes and comet parameters. Rats treated with vitamin C showed a significant increase in absolute and relative epididymis weight compared to the control group. Moreover, vitamin C intervention reversed the adverse effects observed in rats fed hydroxyurea, indicating that low doses of vitamin C can protect against testicular damage.