Kidney Samples Research Articles

The impact of normothermic and hypothermic preservation methods on kidney lipidome—comparative study using chemical biopsy with microextraction probes

IntroductionNormothermic ex vivo kidney perfusion (NEVKP) is designed to replicate physiological conditions to improve graft outcomes. A comparison of the impact of hypothermic and normothermic preservation techniques on graft quality was performed by lipidomic profiling using solid-phase microextraction (SPME) chemical biopsy as a minimally invasive sampling approach.MethodsDirect kidney sampling was conducted using SPME probes coated with a mixed-mode extraction phase in a porcine autotransplantation model of the renal donor after cardiac death, comparing three preservation methods: static cold storage (SCS), NEVKP, and hypothermic machine perfusion (HMP). The lipidomic analysis was done using ultra-high-performance liquid chromatography coupled with a Q-Exactive Focus Orbitrap mass spectrometer.ResultsChemometric analysis showed that the NEVLP group was separated from SCS and HMP groups. Further in-depth analyses indicated significantly (p < 0.05, VIP > 1) higher levels of acylcarnitines, phosphocholines, ether-linked and longer-chain phosphoethanolamines, triacylglycerols and most lysophosphocholines and lysophosphoethanolamines in the hypothermic preservation group. The results showed that the preservation temperature has a more significant impact on the lipidomic profile of the kidney than the preservation method’s mechanical characteristics.ConclusionHigher levels of lipids detected in the hypothermic preservation group may be related to ischemia-reperfusion injury, mitochondrial dysfunction, pro-inflammatory effect, and oxidative stress. Obtained results suggest the NEVKP method’s beneficial effect on graft function and confirm that SPME chemical biopsy enables low-invasive and repeated sampling of the same tissue, allowing tracking alterations in the graft throughout the entire transplantation procedure.

Integrated Proteomics Analysis of Baseline Protein Expression in Pig Tissues.

The availability of an increasingly large amount of public proteomics data sets presents an opportunity for performing combined analyses to generate comprehensive organism-wide protein expression maps across different organisms and biological conditions. Sus scrofa, a domestic pig, is a model organism relevant for food production and for human biomedical research. Here, we reanalyzed 14 public proteomics data sets from the PRIDE database coming from pig tissues to assess baseline (without any biological perturbation) protein abundance in 14 organs, encompassing a total of 20 healthy tissues from 128 samples. The analysis involved the quantification of protein abundance in 599 mass spectrometry runs. We compared protein expression patterns among different pig organs and examined the distribution of proteins across these organs. Then, we studied how protein abundances were compared across different data sets and studied the tissue specificity of the detected proteins. Of particular interest, we conducted a comparative analysis of protein expression between pig and human tissues, revealing a high degree of correlation in protein expression among orthologs, particularly in brain, kidney, heart, and liver samples. We have integrated the protein expression results into the Expression Atlas resource for easy access and visualization of the protein expression data individually or alongside gene expression data.

A Comprehensive Study to Determine the Residual Elimination Pattern of Major Metabolites of Amoxicillin–Sulbactam Hybrid Molecules in Rats by UPLC–MS/MS

Amoxicillin and sulbactam are widely used in animal food compounding. Amoxicillin–sulbactam hybrid molecules are bicester compounds made by linking amoxicillin and sulbactam with methylene groups and have good application prospects. However, the residual elimination pattern of these hybrid molecules in animals needs to be explored. In the present study, the amoxicillin–sulbactam hybrid molecule (AS group) and a mixture of amoxicillin and sulbactam (mixture group) were administered to rats by gavage, and the levels of the major metabolites of amoxicillin, amoxicilloic acid, amoxicillin diketopiperazine, and sulbactam were determined by UPLC–MS/MS. The residue elimination patterns of the major metabolites in the liver, kidney, urine, and feces of rats in the AS group and the mixture group were compared. The results showed that the total amount of amoxicillin, amoxicilloic acid, amoxicillin diketopiperazine, and the highest concentration of sulbactam in the liver and kidney samples of the AS group and the mixture group appeared at 1 h after drug withdrawal. Between 1 h and 12 h post discontinuation, the total amount of amoxicillin, amoxicilloic acid, and amoxicillin diketopiperazine in the two tissues decreased rapidly, and the elimination half-life of the AS group was significantly higher than that in the mixture group (p < 0.05); the residual amount of sulbactam also decreased rapidly, and the elimination half-life was not significantly different (p > 0.05). In 72 h urine samples, the total excretion rates were 60.61 ± 2.13% and 62.62 ± 1.73% in the AS group and mixture group, respectively. The total excretion rates of fecal samples (at 72 h) for the AS group and mixture group were 9.54 ± 0.26% and 10.60 ± 0.24%, respectively. These results showed that the total quantity of amoxicillin, amoxicilloic acid, and amoxicillin diketopiperazine was eliminated more slowly in the liver and kidney of the AS group than those of the mixture group and that the excretion rate through urine and feces was essentially the same for both groups. The residual elimination pattern of the hybrid molecule in rats determined in this study provides a theoretical basis for the in-depth development and application of hybrid molecules, as well as guidelines for the development of similar drugs.

Ultrastructure expansion microscopy (U-ExM) of mouse and human kidneys for analysis of subcellular structures.

Ultrastructure expansion microscopy (U-ExM) involves the physical magnification of specimens embedded in hydrogels, which allows for super-resolution imaging of subcellular structures using a conventional diffraction-limited microscope. Methods for expansion microscopy exist for several organisms, organs, and cell types, and used to analyze cellular organelles and substructures in nanoscale resolution. Here, we describe a simple step-by-step U-ExM protocol for the expansion, immunostaining, imaging, and analysis of cytoskeletal and organellar structures in kidney tissue. We detail the critical modified steps to optimize isotropic kidney tissue expansion, and preservation of the renal cell structures of interest. We demonstrate the utility of the approach using several markers of renal cell types, centrioles, cilia, the extracellular matrix, and other cytoskeletal elements. Finally, we show that the approach works well on mouse and human kidney samples that were preserved using different fixation and embedding conditions. Overall, this protocol provides a simple and cost-effective approach to analyze both preclinical and clinical renal samples in high detail, using conventional lab supplies and standard widefield or confocal microscopy.

184 The metabolism of vitamin D is modulated by vitamin C supplementation during porcine reproductive and respiratory syndrome challenge in post-weaning piglets

Abstract This study evaluated the impact of different dietary vitamin D sources combined or not with vitamin C on serum 25(OH)D3 concentrations and the expression of genes related to antimicrobial peptides and to the metabolism of vitamin D, vitamin C, calcium (Ca), and phosphorus (P). From weaning (d 21) until d 68, 27 piglets were fed a post-weaning diet supplemented with: 1,500 IU of vitamin D3 (CTR; n = 9); 200 IU of vitamin D3 + 2,000 IU of 25(OH)D3 (HD; n = 8); or HD + 500 mg/kg of vitamin C (HD+C; n = 10). From d 40 to 68 piglets were exposed to porcine reproductive and respiratory syndrome (PRRS) using a natural disease challenge model. At d 68, after blood collection, all piglets were euthanized for the collection of lung and kidney samples. The concentrations of serum 25(OH)D3 were determined by HPLC and the mRNA expression of genes of interest was assessed by qPCR. Data were analyzed in a one-way analysis of variance with three treatments. At d 40 (after 19 d of supplementation), serum 25(OH)D3 concentrations were greatest for HD+C (22.48 ± 1.25 ng/mL), intermediate for HD (17.43 ± 0.84 ng/mL), and least for CTR (8.54 ± 0.65 ng/mL) treatments (P < 0.01). At d 68 (after 28 d of PRRS challenge) these values were greatest for HD (14.64 ± 2.77 ng/mL), intermediate for CTR (6.22 ± 0.59 ng/mL), and least for HD+C (2.18 ± 0.13 ng/mL; P < 0.01). No treatments effect was detected on pulmonary CYP27B1, RXRα, SLC23A1, RIGI, PR-39, and pBD1. In lungs, VDR expression was least for HD+C (P ≤ 0.06) whereas MDA5 expression was greatest for HD+C (P ≤ 0.07), and SLC23A2 expression was greatest for HD (P ≤ 0.09; Table 1). No treatments effect was detected on expression of renal FGFR1, KL, RXRα, SLC23A1, SLC23A2, and TPRV5. In kidney, CYP27B1 expression was greatest for HD+C groups (P = 0.03) and CYP24A expression was greatest for HD (P = 0.01). Renal EGR1 and NPT2c expressions were greatest for CTR groups and least for HD+C (P = 0.09) whereas WNK4 expression was greatest for HD+C and least for CTR (P = 0.10). Renal VDR expression was greatest for HD groups and least for HD+C (P ≤ 0.05). In conclusion, vitamin C supplementation increased serum 25(OH)D3 concentrations in pre-challenged piglets. These concentrations were reduced by vitamin C supplementation after PRRS challenge, which may be related to a greater synthesis of the active form of vitamin D in the kidney. Little effects were detected on pulmonary antimicrobial peptides, but the gene expression related to renal P reabsorption was influenced by HD+C whereas Ca reabsorption was apparently not impacted by treatments.

262 Modulation of vitamin D, calcium and phosphorous metabolism and immune response in pre- and post-weaning piglets under different vitamin D supplementation strategies

Abstract This study evaluated the impact of different vitamin D supplementation strategies to pre- and post-weaning piglets on vitamin D, calcium and phosphorous metabolism and immune status. Sixty piglets were selected at d 2 of age [body weight (BW) = 1.71 ± 0.03 kg] and orally supplemented with saline (CTR) or a solution of 25(OH)D3 at d 2, d 8, and d 21 (0.1, 0.2 and 0.3 mg, respectively) along with UVB exposure every second day from d1 4 up to d 21 (HD). From weaning (d 21) until d 42, CTR animals were supplemented with 2,000 UI of vitamin D3/kg of feed and HD animals were supplemented with 2,000 UI 25(OH)D3/kg. Piglets were sacrificed at d 21, d 28, and d 35 (n = 10/treatment/d) for the collection of blood, jejunum, liver, and kidney samples. Serum 25(OH)D3 concentrations were determined by HPLC and the mRNA expression of genes implicated in vitamin D, calcium and phosphorous metabolism and immune status were assessed by quantitative real-time PCR. Data were analyzed using a 2x3 factorial design. Serum 25(OH)D3 concentrations were 2.8, 3.6, and 2.2-fold greater in HD piglets at d 21, 28, and 35 (P ≤ 0.01), respectively (Table 1). No treatment or treatment x time interaction was detected on the expression of genes related to vitamin D metabolism in the jejunum mucosa and liver (P ≥ 0.19). In kidney, HD piglets had greater expression of CYP27B1 (P = 0.09) and decreased expression of CYP24A1 (P = 0.06). Renal EGR1 expression was greater in HD piglets (P = 0.01). No treatment effect was detected on the expression of genes directly implicated in phosphate or calcium reabsorption (P ≥ 0.16). No treatment or treatment x time interaction was detected on the mRNA expression of selected genes related to immune response in the jejunum mucosa, liver, and kidney (P ≥ 0.14). Independently of vitamin D treatments, time effects were detected for gene expression. The mRNA expression of CYP27B1 increased and that of CYP24A1 decreased from d 28 to 35 (P ≤ 0.04) in kidney. Genes related to calcium (TRPV5) and phosphate (NPT2c) reabsorption in kidney had reduced mRNA expression on d 35 compared with d 21 (P ≤ 0.01). Overall, the mRNA expression of genes related to immune response in all studied tissues was lower at d 35 compared with d 21 (P ≤ 0.08). Bodyweight at slaughter was not affected by treatments at d 21, 28, and 35 (P ≥ 0.29). In conclusion, pre-weaning vitamin D supplementation enhanced vitamin D status at weaning and the use of dietary 25(OH)D3 supplementation was more effective in increasing the vitamin D status post-weaning. The mRNA expression results suggest that greater vitamin D status at weaning and during the first 2 wk post-weaning do not affect calcium and phosphate reabsorption in kidney and the immune defense of pre- and post-weaning piglets.

A Systematic Review and Meta-Analysis of microRNA Profiling Studies in Chronic Kidney Diseases

Chronic kidney disease (CKD) represents an increasing health burden. Evidence suggests the importance of miRNA in diagnosing CKD, yet the reports are inconsistent. This study aimed to determine novel miRNA biomarkers and potential therapeutic targets from hypothesis-free miRNA profiling studies in human and murine CKDs. Comprehensive literature searches were conducted on five databases. Subgroup analyses of kidney diseases, sample types, disease stages, and species were conducted. A total of 38 human and 12 murine eligible studies were analyzed using Robust Rank Aggregation (RRA) and vote-counting analyses. Gene set enrichment analyses of miRNA signatures in each kidney disease were conducted using DIANA-miRPath v4.0 and MIENTURNET. As a result, top target genes, Gene Ontology terms, the interaction network between miRNA and target genes, and molecular pathways in each kidney disease were identified. According to vote-counting analysis, 145 miRNAs were dysregulated in human kidney diseases, and 32 were dysregulated in murine CKD models. By RRA, miR-26a-5p was significantly reduced in the kidney tissue of Lupus nephritis (LN), while miR-107 was decreased in LN patients’ blood samples. In both species, epithelial-mesenchymal transition, Notch, mTOR signaling, apoptosis, G2/M checkpoint, and hypoxia were the most enriched pathways. These miRNA signatures and their target genes must be validated in large patient cohort studies.

The nephroprotective effect of Quercetin in Cyclophosphamide-induced renal toxicity might be associated with MAPK/ERK and NF-κB signal modulation activity

The present study aimed to examine the protective effect of quercetin (QUE) on cyclophosphamide (CTX)-induced nephrotoxicity. For that purpose, 24 mice were divided into four groups (Control, QUE, CTX, and CTX + QUE). The CTX and CTX + QUE groups received 200 mg/kg of cyclophosphamide on the 1st and 7th days. The QUE and CTX + QUE groups were treated with 50 mg/kg of quercetin daily for 14 days. At the end of the experiment, the animals were sacrificed, and kidney samples were analyzed. The results indicated that CTX leads to severe morphological degenerations and disruption in renal function. Serum BUN, Creatinine, Uric acid, tissue Bax, Caspase 3, TNF-α and IL-1β expression levels were upregulated in the CTX group compared to Control and QUE groups (p < 0.05). Although MAPK/ERK phosphorylation level is not affected in CTX group, there was a significant increase in CTX + QUE group (p < 0.05), but the NF-κB was significantly suppressed in this group (p < 0.01). The RT-qPCR results showed that the cyt-c and the Bax/Bcl-2 ratio mRNA expression folds were upregulated in the CTX group (p < 0.01), which was downregulated in the CTX + QUE group. However, there was a significant difference in the CTX + QUE group compared to the Control and QUE groups (p < 0.01). The findings showed that administering quercetin along with cyclophosphamide alleviated renal injury by regulating apoptotic and inflammatory expression. Moreover, the administration of quercetin and cyclophosphamide could synergistically improve renal function test results, and activate cellular responses, which upmodulate MAPK/ERK phosphorylation and suppression of NF-κB.

Effects of phoxim on antibacterial infection of silver carp

The efficacy of phoxim in treating bacterial sepsis in silver carp is significant, yet its underlying mechanism remains elusive. This study aimed to establish a model of Aeromonas veronii infection in silver carp and subsequently treat the infected fish with 10 μg/L phoxim. Kidney and intestine samples from silver carp were collected for transcriptome analysis and assessment of intestinal microbial composition, with the aim of elucidating the mechanism underlying the efficacy of phoxim in treating bacterial sepsis in silver carp. The results of transcriptome and intestinal microbial composition analysis of silver carp kidney indicated that A. veronii infection could up-regulate the expression of il1β, il6, nos2, ctsl, casp3, et al, which means, signifying that the kidney of silver carp would undergo inflammation, induce apoptosis, and alter the composition of intestinal microorganisms. Phoxim immersion might enhance the energy metabolism of silver carp and change its intestinal microbial composition, potentially elevating the antibacterial infection resistance of silver carp. These findings may contribute to an understanding of how phoxim can effectively treat bacterial sepsis in silver carp.

Dual-specificity phosphatase 26 protects against kidney injury caused by ischaemia-reperfusion through restraint of TAK1-JNK/p38-mediated apoptosis and inflammation of renal tubular epithelial cells

Dual-specificity phosphatase 26 (DUSP26) acts as a pivotal player in the transduction of signalling cascades with its dephosphorylating activity. Currently, DUSP26 attracts extensive attention due to its particular function in several pathological conditions. However, whether DUSP26 plays a role in kidney ischaemia-reperfusion (IR) injury is unknown. Aims of the current work were to explore the relevance of DUSP26 in kidney IR damage. DUSP26 levels were found to be decreased in renal tubular epithelial cells following hypoxia-reoxygenation (HR) and kidney samples subjected to IR treatments. DUSP26-overexpressed renal tubular epithelial cells exhibited protection against HR-caused apoptosis and inflammation, while DUSP26-depleted renal tubular epithelial cells were more sensitive to HR damage. Upregulation of DUSP26 in rat kidneys by infecting adenovirus expressing DUSP26 markedly ameliorated kidney injury caused by IR, while also effectively reducing apoptosis and inflammation. The mechanistic studies showed that the activation of transforming growth factor-β-activated kinase 1 (TAK1)-JNK/p38 MAPK, contributing to kidney injury under HR or IR conditions, was restrained by increasing DUSP26 expression. Pharmacological restraint of TAK1 markedly diminished DUSP26-depletion-exacebated effects on JNK/p38 activation and HR injury of renal tubular cells. The work reported a renal-protective function of DUSP26, which protects against IR-related kidney damage via the intervention effects on the TAK1-JNK/p38 axis. The findings laid a foundation for understanding the molecular pathogenesis of kidney IR injury and provide a prospective target for treating this condition.

Effects of Short-, Medium-, and Long-Term Treatment Using Photobiomodulation Therapy Combined with Static Magnetic Field in Aging Rats

(1) Background: We investigated the detrimental and protective effects of short-, medium, and long-term treatment with different doses of photobiomodulation therapy combined with static magnetic field (PBMT-sMF) during the aging process. (2) Methods: Rats were treated for 15, 30, and 60 weeks with 1, 3, 10, and 30 J of PBMT-sMF or a placebo control. In addition, eight young rats were not subjected to any procedure or treatment and were euthanized at six weeks old. Skin, muscle, bone, kidney, liver, and blood samples were analyzed. (3) Results: No differences between the groups in the morphology of the skin, muscle, and bone was observed. Glutamic pyruvic transaminase levels were increased in the placebo group after 30 and 60 weeks. Glutamic oxaloacetic transaminase levels were also increased in the placebo group after 30 weeks. An increase in creatinine in the PBMT-sMF 3, 10, and 30 J groups compared with that in the young control group was observed. No significant difference in urea levels between the groups was noted. Vascular endothelial growth factor increased in the PBMT-sMF 10 and 30 J groups after 15 weeks of treatment and in the PBMT-sMF 3 J after 60 weeks. Finally, vascular endothelial growth factor decreased in the PBMT-sMF 30 J group after 30 weeks of treatment. (4) Conclusions: PBMT-sMF did not have detrimental effects on the skin, muscle, bone, kidney, or liver after short-, medium-, and long-term treatments in aging rats. In addition, PBMT-sMF may have protective effects on the muscle tissue in aging rats after short- and long-term treatment.

Early diagnosis and management of finish type congenital nephrotic syndrome in a 3-year-old child: A case report

This study describes a 3-year-old non Finnish girl who was diagnosed with Finnish-type nephrotic syndrome (FTNS), a rare kidney illness brought on by NPHS1 gene abnormalities. Even though she had low protein levels and severe swelling when she was three months old, her case of congenital nephrotic syndrome (CNS) was incorrectly identified as minimal change disease (MCD). The standard MCD treatments, cyclosporine and steroids, did not affect her. A genetic test and a kidney sample confirmed the diagnosis of FTNS, a condition that necessitates an early renal transplant for survival. She is awaiting a kidney transplant and was treated with drugs that save proteins to lessen her edema. The study emphasizes the significance of FTNS early and correct diagnosis as well as the requirement for alternate treatments for this difficult illness.

Essential and toxic elements analysis of wild boar tissues from north-eastern Romania and health risk implications

IntroductionThe level of essential minerals in meat is an important factor in human nutrition and health. Meat from responsibly managed wildlife is an alternative raw material with considerable nutritional benefits. Meat from hunted animals has essential and non-essential elements for the human body. It is important to carefully monitor the levels of heavy metals accumulated in the tissues of hunted animals in polluted areas to ensure food safety and environmental contamination. High levels of heavy metals in food and the environment can pose a danger to human health.MethodsThe study aims to investigate the levels of essential mineral elements and heavy metals in the muscle tissue and organs of wild boar harvested through the herd density control plan over the last decade in north-eastern Romania.Results and discussionThe statistical analysis indicates that the age of the animals had a significant impact on Fe, Cu, and Zn levels in Longissimus lumborum. In the kidney a highly significant difference in Fe content by sex, with males showing higher values than females. The sex was shown to significantly influence the Mg levels. However, there are concerns about the accumulation of heavy metals such as Lead (Pb) and Cadmium (Cd), which may hurt the health of game meat consumers in the study area. Cd level it shows significant differences according to both age and sex, with higher concentrations in adults and males. Statistical analysis shows a negative correlation between Fe and Zn concentrations in muscle samples, while a positive correlation was found between Fe and Mn in kidney samples. There was also a positive association between Zn and Cu in muscle samples, but a negative association in kidney samples. Principal component analysis shows significant variation in essential element and heavy metals data between muscle and kidney samples. The loading plot shows a direct correlation between Pb and Cu and between Pb and Cd. However, an opposite correlation also is observed between Cu and Mg, Cd and Mg, and Pb and Mg. HQ (Hazard Quotient) for children compared to adults indicates a potentially higher risk associated with meat consumption among children because children are more vulnerable than adults. We report for the first time, to the best authors’ knowledge, various levels of essential minerals and exceeded maximum admitted level of heavy metals in the muscle tissues and kidneys of Sus scrofa ferus from Romania intended for human consumption, moreover, our findings highlight the need for strict monitoring and implementation of appropriate corrective measures, given the significant percentages of muscle and kidney samples exceeding the allowable limits for two of the most common toxic metals in the environment.

Occurrence of antimicrobial residues in broiler chickens from selected farms and markets in Kano Metropolis, Nigeria

The indiscriminate use of antibiotics in animal meant for human and animal’s consumption lead to occurrence of antibiotic residues which threaten the health of both animals and humans. The aim of the study was to detect the presence of antimicrobial residues in broiler chicken in Kano state, Nigeria. Samples of fresh breast muscle, thigh muscle, liver and kidney were collected from 100 birds and examined using microbial inhibition test and ELISA test to determine the occurrence of antimicrobial residues and Enrofloxacin residues. A questionnaire was also administered to 33 workers and managers of the selected bird’s farms to assess their knowledge on the use of antibiotics and presence of antimicrobial residues. The result of the study revealed that 43% of the studied birds tested positive to antibiotic residue. Kidney samples recorded the highest (43%) residue present while thigh muscles (35%) recorded the least. The result indicated that among the birds detected with positive antibiotic residues, 36.4% were also positive for Enrofloxacin residue. All the respondents were using one or more antibiotic for treatment, prophylaxis or both and to a lesser extent as growth promoters. Majority (55.5%) of the respondents were not aware of the presence of residues in poultry. Most of the respondents (63.7%) have knowledge of withdrawal period but all of them were not observing withdrawal practice before disposing their birds. Doxycycline antibiotic was found to be the most indiscriminately use antibiotic, while erythromycin and chloramphenicol are the least. The study advocates the need for awareness campaigns for the farmers on the correct way of administering the antibiotics with special emphasis on withdrawal periods as well as continuous monitoring of the residues for public health reasons.

Detection of selected antimicrobial residues in red meat and kidney of beef cattle slaughtered at Nekemte municipal abattoir, Ethiopia.

Antibiotics are commonly used drugs in farm animals for therapeutic, prophylactic and diagnosis purposes. As a result of the use of antibiotics in livestock, residues of antibiotics may be present in animal-derived foods, especially in meat. This study aimed at determining the antibiotic residues in cattle slaughtered at Nekemte municipal abattoir and assessing the knowledge, attitude and practice (KAP) level of the community about antibiotic residue in food of cattle origin. A cross-sectional study was conducted on randomly selected kidney and muscle samples slaughtered at Nekemte municipal abattoir. The samples were collected aseptically and analysed using liquid chromatography mass spectrometer. In addition the KAP of cattle handlers, butcher men and meat users were collected using structured questionnaire. The data was analysed by SPSS and intercooled Stata version 7.0, 2001. Results are presented as percentages and frequency distributions in tabular and graphical form. From 120 individuals interviewed, only 25.83% knew about withdrawal time and had limited knowledge on effect of antibiotic residue on human health, whereas about 47.5% did not heard about antibiotic residue in meat. Tetracycline was detected in all the sampled meat 60 (100%), whereas oxytetracycline residues were detected in half 15 (50%) of the samples. In the current study doxycycline, sulphadiazine, penicillin G and enrofloxacin were not detected in all samples. Oxytetracycline residue levels ranged from 0.00 to 463.35µg/kg for the kidney and 0.00 to 354.55µg/kg for muscle samples. About 10% of kidney and 3.33% muscle samples collected had oxytetracycline residues above maximum residue limits. In general, the study revealed that oxytetracycline residues were prevalent among antimicrobial residues analysed from the study area. The study indicated the presence of high antimicrobial residue and hence exposes for antimicrobial resistance of pathogens warranting coordinated effort to mitigate its health effect on the animal and hence human being.

Grainyhead-like 2 Deficiency and Kidney Cyst Growth in a Mouse Model.

The transcription factor Grainyhead-like 2 (GRHL2) plays a crucial role in maintaining the epithelial barrier properties of the renal collecting duct and is essential for osmoregulation. We noticed a reduction in GRHL2 expression in cysts derived from the collecting ducts in kidneys affected by Autosomal Dominant Polycystic Kidney Disease (ADPKD). However, the specific role of GRHL2 in cystic kidney disease remains unknown. The functional role of the transcription factor Grhl2 in the context of cystic kidney disease was examined through analysis of its expression pattern in patient samples with ADPKD and generating a transgenic cystic kidney disease (TCKD) mouse model by overexpressing the human proto-oncogene c-MYC in kidney collecting ducts. Next, TCKD mice bred with collecting duct-specific Grhl2 knockout mice (Grhl2KO). The resulting TCKD-Grhl2KO mice and their littermates were examined by various types of histological and biochemical assays and gene profiling analysis via RNA-seq. A comprehensive examination of kidney samples from patients with ADPKD revealed GRHL2 downregulation in collecting duct-derived cyst epithelia. Comparative analysis of TCKD and TCKD-Grhl2KO mice exhibited that the collecting duct-specific deletion of Grhl2 resulted in markedly aggravated cyst growth, worsened kidney dysfunction, and shortened life span. Furthermore, transcriptomic analyses indicated sequential downregulation of kidney epithelial cyst development regulators (Frem2, Muc1, Cdkn2c, Pkd2, and Tsc1) during cyst progression in kidneys of TCKD-Grhl2KO mice which included presumed direct Grhl2 target genes. These results suggest GRHL2 as a potential progression modifier, especially for cysts originating from collecting ducts.

Overexpression of cyclin F/CCNF as an independent prognostic factor for poor survival in clear cell renal cell carcinoma

Cyclin F (encoded by CCNF gene) has been reported to be implicated in the pathobiology of several human cancers. However, its potential clinical significance in clear cell renal cell carcinoma (ccRCC) remains unknown. The present study aimed to evaluate the potential significance of cyclin F, assessed by immunohistochemical (IHC) staining and molecular (bioinformatics) techniques, as a prognostic marker in ccRCC in relation to clinicopathological features and outcomes. IHC staining was performed using two independent ccRCC tissue array cohorts, herein called tissue macroarray (TMA)_1 and tissue microarray (TMA)_2, composed of 108 ccRCCs and 37 histologically normal tissues adjacent to the tumor (NAT) and 192 ccRCCs and 16 normal kidney samples, respectively. The mRNA expression data were obtained from The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) public datasets, followed by bioinformatics analysis of biological mechanisms underlying prognosis. The relationship between immune cell infiltration level and CCNF expression in ccRCC was investigated using the Tumor Immune Estimation Resource 2.0 (TIMER2) and Gene Expression Profiling Interactive Analysis 2 (GEPIA2). Cyclin F expression was significantly elevated in ccRCC lesions compared to both NAT and normal renal tissues. Likewise, CCNF mRNA was markedly increased in ccRCCs relative to non-cancerous tissues. In all analyzed cohorts, tumors with features of more aggressive behavior were more likely to display cyclin F/CCNF-high expression than low. Furthermore, patients with high cyclin F/CCNF expression had shorter overall survival (OS) times than those with low expression. In addition, multivariable analysis revealed that cyclin F/CCNF-high expression was an independent prognostic factor for poor OS in ccRCC. Enrichment analysis for mechanistically relevant processes showed that CCNF and its highly correlated genes initiate the signaling pathways that eventually result in uncontrolled cell proliferation. CCNF expression was also correlated with immune cell infiltration and caused poor outcomes depending on the abundance of tumor-infiltrating immune cells in ccRCC. Our findings suggest that cyclin F/CCNF expression is likely to have an essential role in ccRCC pathobiology through regulating multiple oncogenic signaling pathways and affecting the tumor immune microenvironment and may serve as prognostic biomarker and promising therapeutic target in ccRCC.

Characterization of Alternative Splicing in High-Risk Wilms' Tumors.

The significant heterogeneity of Wilms' tumors between different patients is thought to arise from genetic and epigenetic distortions that occur during various stages of fetal kidney development in a way that is poorly understood. To address this, we characterized the heterogeneity of alternative mRNA splicing in Wilms' tumors using a publicly available RNAseq dataset of high-risk Wilms' tumors and normal kidney samples. Through Pareto task inference and cell deconvolution, we found that the tumors and normal kidney samples are organized according to progressive stages of kidney development within a triangle-shaped region in latent space, whose vertices, or "archetypes", resemble the cap mesenchyme, the nephrogenic stroma, and epithelial tubular structures of the fetal kidney. We identified a set of genes that are alternatively spliced between tumors located in different regions of latent space and found that many of these genes are associated with the epithelial-to-mesenchymal transition (EMT) and muscle development. Using motif enrichment analysis, we identified putative splicing regulators, some of which are associated with kidney development. Our findings provide new insights into the etiology of Wilms' tumors and suggest that specific splicing mechanisms in early stages of development may contribute to tumor development in different patients.

Metabolites and metabolic pathway analysis of sulfadimidine in carp (Cyprinus carpio) based on UHPLC-Q-orbitrap HRMS

Sulfadimidine (SM2) is an N-substituted derivative of p-aminobenzenesulfonyl structure. This study aimed to analyze the metabolism of SM2 in carp (Cyprinus carpio). The carps were fed with SM2 at a dose of 200 mg/(kg · bw) and then killed. The blood, muscle, liver, kidney, gill, other guts, and carp aquaculture water samples were collected. The UHPLC-Q-Exactive Plus Orbitrap-MS was adopted for determining the metabolites of SM2 in the aforementioned samples. Twelve metabolites, which were divided into metabolites in vivo and metabolites in vitro, were identified using Compound Discoverer software. The metabolic pathways in vivo of SM2 in carp included acetylation, hydroxylation, glucoside conjugation, glycine conjugation, carboxylation, glucuronide conjugation, reduction, and methylation. The metabolic pathways in vitro included oxidation and acetylation. This study clarified the metabolites and metabolic pathways of SM2 in carp and provided a reference for further pharmacodynamic evaluation and use in aquaculture.

Inhibition of HDAC6 with CAY10603 alleviates acute and chronic kidney injury by suppressing the ATF6 branch of UPR

BackgroundHistone deacetylase 6 (HDAC6) inhibitor CAY10603 has been identified as a potential therapeutic agent for the treatment of diabetic kidney disease (DKD). The objective of this study was to investigate the therapeutic effects of CAY10603 in mice with acute kidney injury (AKI) and chronic kidney diseases (CKD). MethodsRenal immunohistology was performed to assess the expression levels of HDAC6 in both human and mouse kidney samples. C57BL/6J mice were intraperitoneal injected with lipopolysaccharide (LPS) to induce AKI; CD-1 mice were fed with adenine diet to induce adenine-nephropathy as CKD model. Serum creatinine, blood urea nitrogen and uric acid were measured to reflect renal function; renal histology was applied to assess kidney damage. Western blot and immunohistology were used to analyze the unfolded protein response (UPR) level. ResultsHDAC6 was significantly upregulated in renal tubular epithelial cells (RTECs) of both AKI and CKD patients as well as mice. In the murine models of AKI induced by LPS and adenine-induced nephropathy, CAY10603 exhibited notable protective effects, including improvement in biochemical indices and pathological changes. In vivo and in vitro studies revealed that CAY10603 effectively suppressed the activation of activating transcription factor 6 (ATF6) branch of UPR triggered by thapsigargin (Tg), a commonly employed endoplasmic reticulum (ER) stressor. Consistent with these findings, CAY10603 also displayed substantial inhibition of ATF6 activation in RTECs from both murine models of LPS-induced AKI and adenine-induced nephropathy. ConclusionsCollectively, these results suggest that CAY10603 holds promise as a potential therapeutic agent for both acute and chronic kidney injury.