Kidney Tissue Research Articles

Erythropoietin Reduces Inflammation, Oxidative Stress, and Apoptosis in a Rat Model of Bleomycin-Induced Idiopathic Pulmonary Fibrosis.

Idiopathic pulmonary fibrosis (IPF) is a lethal interstitial disease with unknown etiology and no effective cure, posing a great health burden to society. Erythropoietin (EPO) has been demonstrated to have protective roles in various tissues such as brain, spinal cord, heart, kidney and lung tissues. In this study, we investigate the specific anti-inflammatory, antioxidant and antiapoptotic effects of erythropoietin on lung tissue in a bleomycin-induced rat model of idiopathic pulmonary fibrosis. Recombinant human EPO or saline was injected, and the animals were monitored for 14 days after bleomycin instillation. Their hematocrit and serum EPO levels were determined. Histological and immunohistochemical analyses were performed. The extent of tissue injury, determined through morphometric analysis, was significantly decreased in size in animals treated with erythropoietin. An immunohistochemical analysis of the expression of cyclooxygenase-2 (COX-2), inducible synthase of nitric oxide (i-NOS), metalloproteinase-9 (MMP-9), erythropoietin receptor (EPO-R), and cytochrome-C (cyt-C) found these enzymes to be decreased in a statistically significant manner in animals treated with erythropoietin when compared to a non-treated group. The reduced expression of COX-2, i-NOS, MMP-9, EPO-R, and i-NOS in the lung tissues of animals treated with EPO indicates the anti-inflammatory, antioxidant and antiapoptotic action of erythropoietin, suggesting its potential therapeutic role in pulmonary fibrosis.

Exploring immune response toward transplanted human kidney tissues assembled from organoid building blocks

The increasing scarcity of organs and the significant morbidity linked to dialysis require the development of engineered kidney tissues from human-induced pluripotent stem cells. Integrative approaches that synergize scalable kidney organoid differentiation, tissue biomanufacturing, and comprehensive assessment of their immune response and host integration are essential to accomplish this. Here, we create engineered human kidney tissues composed of organoid building blocks (OBBs) and transplant them into mice reconstituted with allogeneic human immune cells. Tissue-infiltrating human immune cells are composed of effector Tcells and innate cells. This immune infiltration leads to kidney tissue injury characterized by reduced microvasculature, enhanced kidney cell apoptosis, and an inflammatory gene signature comparable to kidney organ transplant rejection in humans. Upon treatment with the immunosuppressive agent rapamycin, the induced immune response is greatly suppressed. Our model is a translational platform to study engineered kidney tissue immunogenicity and develop therapeutic targets for kidney rejection.

Establishment and characterization of a novel cell line derived from spotted sea bass Lateolabrax maculatus and its susceptibilities to iridoviruses

The spotted sea bass (Lateolabrax maculatus), which holds significant commercial value, is extensively cultivated in China, Japan, and Southeast Asian countries. This study reports the establishment and characterization of a novel continuous cell line, designated as LMK cells, derived from the kidney tissue of spotted sea bass, The LMK cell line, primarily composed of epithelial-like cells, has been successfully subcultured in Medium 199 supplemented with 10 % fetal bovine serum (FBS) for over 60 passages. Mitochondrial genome sequencing and cytochrome c oxidase subunit I gene amplification verified the origin of the LMK cells from L. maculatus, distinguishing them from L. japonicus. Optimal culture conditions for LMK cells at passage 35 were determined to be 10 % FBS at 27 °C. Chromosomal analysis indicated that the cell line possesses 48 chromosomes. To assess viral susceptibility, LMK cells were exposed to the red sea bream iridovirus and largemouth bass ranavirus. Both iridoviruses induced cytopathic effects within 3 days post-infection and triggered immune-related gene responses. This study demonstrates that the LMK cell line is a valuable resource for molecular, virological, and immunological research in L.maculatus and other sea bass species.

Trans-cinnamic acid alleviates high-fat diet-induced renal injury via JNK/ERK/P38 MAPK pathway

Obesity-related chronic kidney disease (CKD) poses a significant risk to individuals' health and wellbeing, but the pathological mechanisms and treatment strategies are currently limited. Trans-cinnamic acid (CA) is a key active monomer found in cinnamon bark and is known for its diverse pharmacological activities. However, its effect on obesity-related renal injury remains unknown. In the current study, the in vitro and in vivo experiments were combined to investigate the beneficial effect of CA on renal injury induced by HFD or PA. We found that CA significantly reduced the obesity of zebrafish body and the accumulation of fat in kidney tissues. The histopathological changes and dysfunction induced by HFD were effectively mitigated by CA administration, as evidenced by the detection of Hematoxylin-Eosin straining, NAG activity, creatinine level, and expression of functional-related genes, respectively. Additionally, the in vitro and in vivo findings demonstrated that CA dramatically reduced the oxidative stress, inflammatory, and apoptosis in HFD-induced kidney tissues or PA-treated HEK293T and HK-2 cells. Finally, the results regarding ERK, JNK, and P38 proteins phosphorylation confirmed that CA may alleviate HFD-induced renal injury by inhibiting the phosphorylation of ERK, JNK, and P38 MAPK proteins. This theory was further supported by the results of co-treatment with anisomycin (a JNK activator) or lipopolysaccharide and CA in HEK293T cells. This study proves that CA alleviates the obesity-related CKD probably through inhibition of MAPK signaling pathway.

Quantitative proteomics analysis reveals the protective role of S14G-humanin in septic acute kidney injury using 4D-label-free and PRM Approaches

Mitochondrial dysfunction contributes to septic acute kidney injury (S-AKI), making mitochondrial protection a potential therapeutic strategy. This study investigates the effects of S14G-humanin (HNG) in S-AKI, utilizing 4D-label-free and parallel reaction monitoring (PRM) techniques for proteomic analysis. An S-AKI model was created in male C57BL/6 mice using lipopolysaccharide (LPS) injection, followed by HNG administration. After 24 h, kidney tissues were analyzed for histology, biochemistry, mitochondrial function, and proteomics. HNG treatment improved renal function, reduced tubular injury, and decreased pro-inflammatory cytokines and oxidative stress markers. Proteomic analysis identified 5900 proteins, with 5111 quantifiable. HNG altered the expression of 132 proteins, with 18 selected for PRM validation. Ten of these proteins were linked to key pathways, including fatty acid degradation and PPAR signaling. This study is the first to show HNG's protective effects in S-AKI, providing insights into its mechanisms through advanced proteomic techniques.

Therapeutic Effect of Thymoquinone on Melatonin, Ferritin, and Renal Function in Renal Ischemia/Reperfusion Injury in Rats

Objective: Ischemia/reperfusion (I/R) injury is the period of tissue or organ damage that develops after the tissue's blood flow is restored. The extent of damage varies according to the severity of tissue and cell damage. Thymoquinone (TQ) has a wide therapeutic spectrum. The effect of thymoquinone on melatonin and ferritin in I/R can regulate renal function by combining these two mechanisms to improve damage. Therefore, the effect of thymoquinone on melatonin and ferritin levels in renal I/R as well as its regulatory role in renal functions have been investigated. Methods: Thirty-six male Sprague Dawley rats were included in the study (250-300 g, 8-10 weeks). The rats were randomly assigned to 6 groups with 6 animals in each group. Groups; 1- Control, 2- Sham, 3- Solvent, 4- Renal ischemia/reperfusion injury (I/R), 5- I/R+ Thymoquinone (TQ) (5 mg/kg/day), 6- TQ (5 mg/kg/day). The dorsal region of the rats was surgically opened, and the left renal artery was clamped for 30 minutes and then reperfused for 24 hours. TQ (i.p) was applied to the treatment groups for 15 days. At the end of the experiment, blood samples were taken from all groups, and kidney function tests (Na+, K+, Creatinine, urea, BUN) were performed. Melatonin and ferritin levels were analyzed by the ELISA method from kidney tissue samples. Results: Data showed that short-term TQ treatment was effective on serum K+ (P = 0.010) and melatonin and ferritin levels in kidney tissue. Melatonin and iron activity, which were normal in healthy groups, melatonin decreased and ferritin increased significantly in the I/R group. TQ treatment positively regulated the dysregulation of these two molecules in I/R. Conclusion: TQ may contribute to the healing of the damage by improving the K+ levels, which indicates the insufficiency of kidney functions in I/R damage. Melatonin and ferritin, as interacting molecules in I/R, are regulated by TQ, indicating that they may contribute to the management of I/R damage.

THE EFFECTIVENESS OF STEM CELL THERAPY IN MYOCARDIAL INFARCTION REVIEWED FROM UREUM, CREATININE AND KIDNEY HISTOPATHOLOGY LEVELS

This study aims to study the effectiveness of single placental stem cell therapy or cardiomyocyte combination in pigs experiencing myocardial infarction in terms of urea, creatinine and kidney organ histopathology values. A total of 9 pigs were divided int o three groups: without therapy (K1), single placental stem cell therapy (K2), and cardiomyocyte coculture combination therapy (K3). All pigs underwent circumflex artery ligation of the proximal coronary artery branch to stimulate cardiac ischemia and therapy was given after myocardial infarction and ST-elevation were formed. The pigs were then treated for eight weeks, euthanized and necropsied for kidney organ removal. The re sults showed a significant decrease in urea and creatinine values (P0.05) in K2 and K3 as compared to K1. Histopathology results showed a decrease in glomerular atrophy values (K2: 10.26±0.93 and K3: 12.44±1.21), tubular necrosis (K2: 13.67±3.76 and K3: 17.23±0.55), and tubular dilation (K2: 16.36±1.26 and K3: 18.03±1.76) and these values were significantly different than those observed in K1. It was concluded that the administration of placental stem cell therapy or combined with cardiomyocyte coculture could reduce urea, creatinine values, and improve kidney tissue histopathologically.

Assessment of sub-lethal effects of Celcron on Java barb through erythrocyte morphology and acetylcholinesterase activity: Implications for environmental health in aquatic ecosystems

Industrialization and the extensive use of chemicals have raised significant concerns about their environmental impacts, particularly on aquatic ecosystems. This study evaluated the sub-lethal effects of Celcron (Cec), an organophosphate insecticide, on the Java barb (Barbonymus gonionotus) through erythrocyte morphology and acetylcholinesterase (AChE) activity, aiming to refine biomarkers for environmental health assessments. We hypothesized that sub-lethal Cec exposure would induce significant erythrocyte abnormalities and decrease AChE activity in Java barb, with variable recovery rates between gill and kidney tissues. To test this, we exposed the juvenile Java barbs to two sub-lethal Cec concentrations - 0.01 ppm (10 % of the LC50) and 0.05 ppm (50 % of the LC50) -for 60 days. After the exposure period, the fish were placed in pesticide-free water to allow for recovery. Results indicated a significant decline in AChE activity in both liver and kidney tissues, with activity levels showing gradual recovery over time. Erythrocyte abnormalities, including nuclear and cellular changes, were significantly elevated in response to Cec exposure. The frequency of nuclear abnormalities such as micronuclei and binucleation increased in a concentration- and duration-dependent manner, with the gill blood exhibiting higher sensitivity and slower recovery compared to kidney blood. Cellular abnormalities such as twin, teardrop and spindle-shaped cells were also more prevalent in Cec-treated fish. Recovery from these abnormalities was observed but varied between gill and kidney blood, with gill blood showing higher sensitivity and slower recovery compared to kidney blood. This study underscores the utility of AChE activity and erythrocyte abnormalities as biomarkers for assessing pesticide impacts on aquatic organisms. The findings highlight the sensitivity of fish erythrocytes to environmental contaminants and emphasize the need for continued research to better understand the long-term effects of pesticide exposure on aquatic health and ecosystem stability.

NLRP3 inflammasome activation and macrophage distribution in kidney tissues from patients with acute oxalate nephropathy.

The current study was initiated to evaluate renal nucleotide-binding and oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome pathway activation and macrophage subtype distribution and their clinicopathological significance in a cohort of oxalate-induced acute kidney injury. Twelve patients with biopsy-proven acute oxalate nephropathy (AON) from January 2016 to October 2022 were retrospectively enrolled, with estimated glomerular filtration rate (eGFR)-matched 24 patients with acute tubulointerstitial nephritis (ATIN) as disease control. Pathological lesions as well as markers of NLRP3 inflammasome pathway and macrophage phenotype detected by immunohistochemistry staining were semi-quantitatively analyzed. Oxalate depositions were found in 5% to 20% of tubules with a positive correlation with Sirius Red staining in AON specimens (rp = 0.668, p = 0.02). Disruption of tubular basement membrane and inflammatory cell reaction was more prominent in specimens of AON (both p < 0.05) as compared with ATIN specimens. The expressions of NLRP3, caspase-1, and gasdermin D were significantly increased in AON specimens as well (all p < 0.05). Patients with M1/M2 macrophage ratio <1 were found with more chronic tubulointerstitial lesions and presented with lower eGFR at the last follow-up (24.8  10.6 mL/min/1.73 m2 vs. 55.1  21.2 mL/min/1.73 m2, p = 0.02) in the AON group. The NLRP3 inflammasome pathway was activated in the kidneys of AON patients, and the ratio of M1 and M2 macrophages was associated with chronicity of pathological changes, which needs further exploration.

Punica granatum L. peel extract protects diabetic nephropathy by activating the Nrf-2/HO-1 pathway.

Diabetes raises cardiovascular morbidity and mortality worldwide and causes retinopathy, neuropathy, and nephropathy. Punica granatum L. (Pomegranate) is a fruit that has been used for its medicinal properties in various cultures. This article aims to investigate the antioxidant, anti-inflammatory, anti-apoptotic activity of Punica granatum L. peel ethanol extract (PGE) and its efficacy on NF-κB and Nrf-2/HO-1 signaling pathways in kidney tissue of rats with streptozotocin-induced diabetes. Single dose STZ 60mg/kg/i.p. rats were given to induce diabetes and blood glucose measurements were taken 7 days later. PGE 10mg/kg/p.o. administered to the treatment groups for 20 days. Blood, kidney, and pancreas samples taken from anesthetized rats were analyzed biochemically and histopathologically. It was observed that STZ increased the levels of urea, uric acid and creatine in the blood, while PGE significantly decreased these parameters. The diabetic group had higher MDA and lower renal tissue GSH level, CAT, GPx, and SOD activity than the control group. STZ also enhanced inflammation, apoptosis, Bax, Caspase-3, and NF-κB expression, and decreased Bcl-2, HO-1, and Nrf-2 expression. Experimental results showed that PGE has the potential to alleviate the harmful effects on the kidney and pancreas by altering the mentioned parameters in diabetic rats.

Long-term biological effects after acute 131I-administration of two rat models (with and without thyroid)

Purpose Radioiodine-131 (RAI or iodine-131) is one of the most frequently used radionuclides for diagnosis and therapy of thyroid diseases (90% of all therapies in nuclear medicine). In order to optimize the patient protection, it is important to evaluate the long-term biological effects of RAI therapy on non-target organs. Materials and methods An experimental animal model has been adopted, it consists on miming RAI therapy. An activity of RAI has been administrated in two models of Wistar rats: the first model with an intact thyroid gland (Thy + model), and the second one was thyroidectomized (Thy- model). For each model, 6 rats were orally contaminated with a solution 18.5 ± 1MBq of [131I]NaI and 6 others rats were used as controls. The 24 rats have been placed in individual cages for a period of 08 months then they were euthanized. The blood was collected by cardiac puncture and all organs were immediately removed. A fraction of thyroid, liver, kidneys and testicles was put in vials containing formaldehyde (10%) for histological investigation. Results Histological observations show some liver disorders more accentuated in the case of the Thy- model, the appearance of kidney tissue effects (hemosiderin deposits, fibrosis and glomerular necrosis) for both models and an absence of any anomaly for the testicles slides. The disturbance of blood parameters specific to each organ has been revealed. Conclusions Long-term biological effect of 131I-administration shows the appearance of various histological disorders confirmed by disturbances in hepatic and renal functions.

The identification of key molecules and pathways in the crosstalk of calcium oxalate-treated TCMK-1 cells and macrophage via exosomes

The interplay between crystals and epithelial cells forms the cornerstone of kidney stone development, communication between epithelial cells and macrophages emerging as a pivotal role in this process. We conducted next-generation sequencing on the secreted exosomes of TCMK-1 cells treated with calcium oxalate monohydrate (OX_EXO) or controls (NC_EXO), and on the macrophage cell line RAW264.7 stimulated with OX_EXO or NC_EXO, followed by validation of differentially expressed target proteins and miRNAs through Western blot and PCR. UPSET plots were employed to identify genes co-targeted by exosomal miRNAs. Various bioinformatic analyses were employed to predict potential mechanisms of the dysregulated genes. We integrated sequencing data from the GEO database, and validated findings using clinical patient urine and kidney tissues. We identified 665 differentially expressed exosomal miRNAs between OX_EXO and NC_EXO. Among the top 10 down-regulated miRNAs, the most targeted genes were AAK1 and NUFIP2, whereas PLCB1 was significantly targeted among the top 10 up-regulated miRNAs. In clinical specimens, we confirmed the differential expressions of five homologous miRNAs, as well as CNOT3, CNCNA1C, APEX1, and TMEM199. In conclusion, treatment of TCMK-1 cells with calcium oxalate significantly alerted the expression profile of exosomal miRNAs, subsequently influencing gene expression in macrophages, thereby modulating the processes of kidney stone formation.

Pik3c3 expression profiling in the mouse kidney and its role in proximal tubule cell physiology.

The class 3 phosphatidylinositol 3-kinase (Pik3c3) plays critical roles in regulating autophagy, endocytosis, and nutrient sensing, but its expression profile in the kidney remains undefined. Recently, we validated a Pik3c3 antibody through immunofluorescence staining of kidney tissues from cell type-specific Pik3c3 knockout mice. Immunohistochemistry unveiled significant disparities in Pik3c3 expression levels across various kidney cell types. Notably, renal interstitial cells exhibit minimal Pik3c3 expression. Further, coimmunofluorescence staining, utilizing nephron segment- or cell type-specific markers, revealed nearly undetectable levels of Pik3c3 expression in glomerular mesangial cells and endothelial cells. Intriguingly, although podocytes exhibit the highest Pik3c3 expression levels among all kidney cell types, the renal proximal tubule cells (RPTCs) express the highest level of Pik3c3 among all renal tubules. RPTCs are known to express the highest level of the epidermal growth factor receptor (EGFR) in adult kidneys; however, the role of Pik3c3 in EGFR signaling within RPTCs remains unexplored. Therefore, we conducted additional cell culture studies. The results demonstrated that Pik3c3 inhibition significantly delayed EGF-stimulated EGFR degradation and the termination of EGFR signaling in RPTCs. Mechanistically, Pik3c3 inhibition surprisingly did not affect the initial endocytosis process but instead impeded the lysosomal degradation of EGFR. In summary, this study defines, for the first time, the expression profile of Pik3c3 in the mouse kidney and also highlights a pivotal role of Pik3c3 in the proximal tubule cells. These findings shed light on the intricate mechanisms underlying Pik3c3-mediated regulation of EGFR signaling, providing valuable insights into the role of Pik3c3 in renal cell physiology. NEW & NOTEWORTHY This is the first report defining the class 3 phosphatidylinositol 3-kinase (Pik3c3) expression profile in the kidney. Pik3c3 is nearly absent in renal interstitial cells, glomerular mesangial cells, and endothelial cells. Remarkably, glomerular podocytes express the highest Pik3c3 level in the kidney. However, the proximal tubule exhibits the highest expression level among all renal tubules. This study also unveils the pivotal role of Pik3c3 in regulating EGFR degradation and signaling termination in RPTCs, furthering our understanding of Pik3c3 in renal cell physiology.

C4d Is an Independent Predictor of the Kidney Failure in Primary IgA Nephropathy.

Background: C4d deposits are present in a substantial proportion of patients with IgA nephropathy (IgAN), indicating the activation of the lectin pathway (LP) of the complement system. It seems that patients with activated LP have worse renal prognosis. The aim of this study was to investigate the prevalence and prognostic significance of C4d in our cohort of patients with primary IgA nephropathy (pIgAN). Methods: Patients with pIgAN were recruited from a hospital register of kidney biopsies of the Department of Nephrology and Dialysis, Dubrava University Hospital, Zagreb. Additional immunohistochemistry staining for C4d was performed on paraffin-embedded kidney tissue, and patients were stratified into being C4d positive or C4d negative. The clinical and histologic features of patients were analyzed and compared regarding C4d positivity. The primary outcome was defined as kidney failure (KF), and predictor variables of KF and renal survival were analyzed. Results: Of a total of 95 patients with pIgAN included in the study, C4d was present in 43 (45.3%). C4d-positive patients had a higher value of systolic (p = 0.039) and diastolic (p = 0.006) blood pressure at diagnosis as well as higher 24 h proteinuria (p = 0.018), serum urate (p = 0.033), and lower eGFR (p < 0.001). C4d-positive patients had worse renal survival (p < 0.001), higher rates of disease progression to KF (p < 0.001), and higher proteinuria (p < 0.001) and lower eGFR (p < 0.001) at the last follow-up. Glomerular C4d was an independent predictor of disease progression to KF (HR = 5.87 [0.95 CI 1.06-32.44], p = 0.032). Conclusions: C4d is an independent predictor of disease progression in patients with pIgAN. C4d may be used as an additional marker of progressive disease course in IgAN. The therapeutic implications of C4d status in IgAN, particularly in terms of complement inhibitors application, are not yet known.

Genomic and transcriptomic landscape to decipher the genetic basis of hyperpigmentation in Lanping black-boned sheep (Ovis aries)

BackgroundLanping black-boned sheep (LPB) represent a distinctive mammalian species characterized by hyperpigmentation, resulting in black bone and muscle features, in contrast to their conventional counterparts exhibiting red muscle and white bone. The genetic basis underlying LPB hyperpigmentation has remained enigmatic.MethodsIn this study, we conducted whole-genome sequencing of 100 LPB and 50 Lanping normal sheep (LPN), and integrated this data with 421 sequenced datasets from wild and domestic sheep, shedding light on the genetic backdrop and genomic variations associated with LPB. Furthermore, we performed comparative RNA-Seq analysis using liver sample to pinpoint genes implicated in the pigmentation process. We generated a comprehensive dataset comprising 97,944,357 SNPs from 571 sheep, facilitating an in-depth exploration of genetic factors.ResultsPopulation genetic structure analysis revealed that the LPB breed traces its origin back to LPN, having evolved into a distinct breed. The integration of positively selected genes with differentially expressed genes identified two candidates, ERBB4 and ROR1, potentially linked to LPB hyperpigmentation. Comparative analysis of ERBB4 and ROR1 mRNA relative expression levels in liver, spleen, and kidney tissues of LPB, in comparison to Diqing sheep, revealed significant upregulation, except for ERBB4 in the liver. Gene expression heatmaps further underscored marked allelic frequency disparities in different populations.ConclusionOur findings establish the evolutionary lineage of the LPB breed from LPN and underscore the involvement of ERBB4 and ROR1 genes in melanin synthesis. These results enhance our comprehension of the molecular basis of hyperpigmentation and contribute to a more comprehensive depiction of sheep diversity.

Investigation of the effects of apilarnil and imatinib use on liver and kidney tissues in rats via PI3K/AKT/mTOR and JAK2/STAT3 pathways

Investigation of the effects of apilarnil and imatinib use on liver and kidney tissues in rats via PI3K/AKT/mTOR and JAK2/STAT3 pathways

Acute and sub-chronic toxicity studies of methanol extract of Trema orientalis (Linn) blume in albino wistar rats

BackgroundThe safety potential of the methanol extract of Trema orientalis (TOM) leaf was evaluated in albino Wistar rats using biochemical, haematological, and histological indices in both acute and sub-chronic toxicity studies. MethodsThe animals were managed following the National Institute of Health (NIH) stipulated protocols for handling laboratory animals. The weight of each animal was recorded upon arrival and monitored throughout the study. The animals were allowed to acclimatize for 14 days, after which they were reweighed and randomly distributed into four groups of three female rats (n=12) for acute toxicity studies. Group A was given distilled water, and Groups B, C, and D were given a single dose of 2000, 4000, and 5000 mg/kg bw TOM extract, respectively. On day 15, each animal was anaesthetized and then euthanized. For the sub-chronic toxicity study, animals were randomly distributed into five groups of eight female rats (n=40). They were dosed daily for 28 days. Group A (Negative control group) was given distilled water. Groups B, C, and D had 200, 400, and 800 mg/kg bw TOM extract and Group E (Vehicle control group) were given 0.25 % of sodium carboxyl methyl cellulose (CMC). Five animals were anaesthetized and then euthanized on day 29, while three animals were kept for recovery evaluation for another two weeks without further administration of the extract. Ten organs were excised from each animal and weighed. The liver and kidney were processed for histopathological studies, while the blood samples were collected for biochemical and haematological assays. ResultsFrom acute toxicity studies, the LD50 value of TOM extract was estimated to exceed 5000 mg/kg bw via oral passage. From Sub-chronic toxicity studies, biochemical results showed a significant (p < 0.05) reduction in total protein, albumin, globulin, AST, ALP, and ALT in a dose-dependent manner. Histology of the liver and kidney tissues of all the animals except the kidney of the 800 mg/kg group had no visible lesions; this sets the safety dose for TOM at above 400 mg/kg but below 800 mg/kg. Recovery animals had significantly (p < 0.05) increased total protein, total bilirubin, and ALP and decreased albumin and direct bilirubin levels. ConclusionThis study reports the safety dose of TOM, a reputable medicinal plant extract. This is the first study reporting that the LD50 value of TOM extract exceeds 5000 mg/kg bw via oral passage.

Prevention of Transition from Acute Kidney Injury to Chronic Kidney Disease Using Clinical-Grade Perinatal Stem Cells in Non-Clinical Study.

Acute kidney injury (AKI) is widely recognized as a precursor to the onset or rapid progression of chronic kidney disease (CKD). However, there is currently no effective treatment available for AKI, underscoring the urgent need for the development of new strategies to improve kidney function. Human placental mesenchymal stromal cells (hpMSCs) were isolated from donor placentas, cultured, and characterized with regard to yield, viability, flow cytometry, and potency. To mimic AKI and its progression to CKD in a rat model, a dedicated sensitive non-clinical bilateral kidney ischemia-reperfusion injury (IRI) model was utilized. The experimental group received 3 × 105 hpMSCs into each kidney, while the control group received IRI and saline and the untreated group received IRI only. Urine, serum, and kidney tissue samples were collected over a period of 28 days. The hpMSCs exhibited consistent yields, viability, and expression of mesenchymal lineage markers, and were also shown to suppress T cell proliferation in a dose-dependent manner. To ensure optimal donor selection, manufacturing optimization, and rigorous quality control, the rigorous Good Manufacturing Practice (GMP) conditions were utilized. The results indicated that hpMSCs increased rat survival rates and improved kidney function by decreasing serum creatinine, urea, potassium, and fractionated potassium levels. Furthermore, the study demonstrated that hpMSCs can prevent the initial stages of kidney structural fibrosis and improve kidney function in the early stages by mitigating late interstitial fibrosis and tubular atrophy. Additionally, a robust manufacturing process with consistent technical parameters was established.

In vitro and in vivo evaluation of hypoglycemic potential and acute toxicity of microencapsulated turmeric instant powder

BackgroundCurcuma longa L. (turmeric), traditionally used to treat various ailments including diabetes, has demonstrated beneficial health effects. However, its potential in food formulations requires further investigation. PurposeThis study examined the hypoglycemic effects of an instant powder containing microencapsulated turmeric oleoresin both in vitro and in vivo, alongside an acute toxicity assessment in Holtzman rats. MethodsThis study encompassed both in vitro and in vivo evaluations. In vitro, the α-glucosidase (a digestive enzyme) inhibition assay was employed to determine the powder's effectiveness. In vivo assessments included: (1) evaluation of the hypoglycemic effect in mice; (2) conducted safety testing following international guidelines, administering a 2000 mg/kg dose to rats with subsequent monitoring of biochemical parameters; and (3) histological examination of liver, kidney, and spleen tissues for potential harmful effects. ResultsThe instant powder with turmeric oleoresin microencapsulation demonstrated significant α-glucosidase inhibition in vitro and hypoglycemic effects in vivo. The acute toxicity test at 2000 mg/kg showed no mortality but revealed significant alterations in erythrocyte parameters and hemoglobin percentage. Histological examinations of liver, kidney, and spleen tissues did not differ from those of the control group, though pancreatic tissue was not evaluated in this study. ConclusionAccording to this study, the microencapsulated turmeric oleoresin instant powder exhibited antidiabetic effects without signs of overt toxicity at doses up to 2000 mg/kg in rats.

Oral administration of recombinant Lactococcus lactis expressing largemouth bass (Micropterus salmoides) IFNa3 protein enhances immune response against largemouth bass virus (LMBV) infection

Largemouth bass virus (LMBV) is a highly pathogenic pathogen that often causes high mortality of affected largemouth bass and significant financial losses. Type I interferon as an effective and broad spectrum tool has been successfully used for therapeutic or prophylactic treatment some viral infections. However, the implementation of immunotherapies based on interferon administration to combat LMBV infections has not been reported. And Lactic Acid Bacteria (LAB) are a powerful vehicle for expressing cytokines or immunostimulant peptides at the gastrointestinal level after oral administration. In this study, Lactococcus lactis (L. lactis) expression system with lactose as a screening marker was utilized to express the Micropterus salmoides interferon a3 (IFNa3) protein and orally administered to largemouth bass. The genetically engineered strain pNZ8149-Usp45-IFNa3-6His/L. lactis NZ3900 was successfully constructed, and its potential to elicit immune protection response by oral administration was evaluated. After orally administration, the recombinant L. lactis was detected in guts of experimental fish and remained detectable for 72 h. Additionally, IFNa3 was able to enhance the test fish's immune response, as determined by the relatively increased mRNA relative expression of immune-related genes in the liver, spleen, and kidney tissues, including IFN-γ, TNF-α, IL-1β, IL-8, IgM and IgT. Following LMBV challenge, the experiment group of pNZ8149-Usp45-IFNa3-6His/L. lactis NZ3900 exhibited a 70 % survival rate, while survival rate were 15 % in the PBS control group, 45 % in the pNZ8149/L. lactis NZ3900 group. Furthermore, the viral load in the surviving fish was significantly lower than that of the control groups. These findings suggest that oral administration of recombinant L. lactis producing IFNa3 induces largemouth bass immune responses at a systemic level to effective prevent and combat of LMBV infection.