Kidney Samples Research Articles

DIAGNOSTIC INVESTIGATION OF AMERICAN ROBINS (TURDUS MIGRATORIUS) WITH FATAL ETHYLENE GLYCOL TOXICOSIS AND CONCURRENT PLASMODIUM SPP. INFECTION WITH RETROSPECTIVE REVIEW OF 44 YEARS OF DIAGNOSTIC DATA.

We describe a mortality event involving at least 44 adult American robins (Turdus migratorius) that were found dead on the morning of February 25, 2023 in a residential backyard in Natchitoches Parish, Louisiana. Five carcasses were submitted for diagnostic evaluation. All five robins were in good nutritional and feather condition with no external indicators of injury or illness. Grossly, kidneys were diffusely pale in all robins and 4/5 also had splenomegaly. Consistent histologic lesions included severe, multifocal to coalescing, renal tubular degeneration and necrosis with intraluminal, birefringent calcium oxalate crystals and calcified concretions. Toxicologic testing revealed a markedly elevated calcium concentration (10,000 parts per million) in a pooled kidney sample. Collectively, these findings support a diagnosis of ethylene glycol toxicosis. Ethylene glycol, an ingredient in antifreeze, is a hazardous toxicant in domestic and wild animals living in residential areas, although confirmed reports in songbirds are scarce. In the present case, the circumstances of ethylene glycol exposure are unknown but most likely anthropogenic in origin. Additionally, splenomegaly prompted molecular testing, which detected Plasmodium unalis in three robins and P. vaughani in one robin. Both Plasmodium spp. can infect a wide range of hosts, and although infections likely were subclinical, their detection raises awareness of their potential pathogenicity in wild birds. This mortality event underscores the need to recognize health risks posed by environmental contaminants and vector-borne pathogens to wildlife, and reinforces that wildlife, especially peridomestic species, can serve as sentinels for risks to domestic animal and human health.

Attenuation of inflammation, oxidative stress and TGF-β1/Smad3 signaling and upregulation of Nrf2/HO-1 signaling mediate the protective effect of diallyl disulfide against cadmium nephrotoxicity

Heavy metals are toxic environmental pollutants with serious health effects on humans and animals. Cadmium (Cd) is known for its serious nephrotoxic effect and its toxicity involves oxidative stress (OS) and inflammation. Diallyl disulfide (DADS), a main constituent of garlic, exhibited cytoprotective and antioxidant activities. This study investigated the effect of DADS on OS, inflammation, and fibrosis induced by Cd in rat kidney, pointing to the involvement of transforming growth factor-β (TGF-β)/Smad3 and nuclear factor erythroid 2–related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling, and peroxisome proliferator-activated receptor gamma (PPARγ). Rats received DADS for 14 days and Cd on day 7 and blood and kidney samples were collected. Cd elevated serum creatinine, urea and uric acid, provoked kidney histopathological alterations and collagen deposition, increased kidney malondialdehyde (MDA) level, and decreased glutathione (GSH) and antioxidant enzymes. Nuclear factor-kappaB (NF-κB) p65, interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-1β, and CD68 were upregulated in Cd-administered rat kidney. DADS prevented kidney injury, mitigated OS, suppressed NF-κB, CD68 and pro-inflammatory mediators, and boosted antioxidants. DADS downregulated TGF-β1, Smad3 phosphorylation and Kelch-like ECH-associated protein-1 (Keap1), and increased Nrf2, HO-1, cytoglobin, and PPARγ. In conclusion, DADS protects the kidney against Cd toxicity by attenuating OS, inflammation, and TGF-β1/Smad3 signaling, and enhancement of Nrf2/HO-1 signaling, antioxidants, and PPARγ.

Depletion of intrinsic renal macrophages with moderate-to-high expression of CD163, MRC1, PTH2R, PDE4D, and CUBN in regulating podocyte injury in diabetic nephropathy: a single-cell RNA sequencing analysis.

Diabetic nephropathy (DN), a severe complication of diabetes, is characterized by glomerular and tubular damage, which often leads to end-stage renal disease (ESRD). The role of renal macrophages (Mφs), particularly their phenotypic plasticity and function in DN, remains poorly understood. This study investigated the key factors influencing Mφ polarization and their impact on podocyte (PODO) injury in DN. Single-nuclear RNA sequencing (snRNA-seq) data from DN and control (CON) kidney samples were analyzed for cell clustering, differential expression, and cell communication. Mφs were identified and categorized based on their gene expression profiles. The proportions and functions of different Mφ phenotypes were compared between DN and CON samples, with a focus on their interaction with PODOs. A subset of Mφs, characterized by high expression of CD163, MRC1, PTH2R, PDE4D, and CUBN, was significantly depleted in DN as compared to in CON samples. This depletion was associated with the overexpression of AHR and underexpression of IGF1R, inhibiting the differentiation of these protective Mφs. The remaining Mφs in the DN samples exhibited altered functions, particularly in regulating oxidative stress and tight junctions. Their interaction with PODOs through ligands including NRG3 and THBS1 suggested a role in promoting PODO dysfunction and apoptosis and their contribution to the progression of DN. The depletion of Mφs with a moderate-to-high expression of CD163, MRC1, PTH2R, PDE4D, and CUBN in patients with DN leads to enhanced PODO injury and apoptosis, highlighting a potential therapeutic target for mitigating DN progression. Further research into the mechanisms governing Mφ-PODO interactions could provide insights into novel treatment strategies for DN.

Dexamethasone: a cheap and convenient way to alleviate subsequent stenosis after thermal injury on ureter.

Ureteral thermal injury (UTI) occurs when the ureter is exposed to high temperatures during surgical procedures, leading to tissue damage and ureteral stricture. Dexamethasone (DEX) has been widely used to treat a variety of diseases for its anti-inflammatory and immunosuppressant effects. However, its role in treating UTI remains unclear. The aim of this study was to determine the efficacy of treatment in mice with UTI. Thirty C57BL/6J mice were randomly divided into three groups (n=10 per group): (I) control group, (II) UTI group, (III) UTI + DEX group. UTI mouse models were constructed using a fast-pack pro-obturation pack handpiece instrument, and the therapeutic effect of DEX (2.5 mg/kg) on UTI was investigated via intraperitoneal injection for two weeks. Pathological changes in the ureter, renal function, extracellular matrix protein deposition, and inflammatory markers in the kidneys were evaluated in mouse models. Ureteral histopathological changes were more severe in the UTI group than in the control group; however, DEX treatment alleviated these changes to a certain extent. Mice in the UTI group exhibited elevated renal function. Although DEX alleviated renal function deterioration, the difference was not statistically significant. Moreover, in comparison to the control group, kidney samples from the mice in the UTI group revealed increased messenger RNA (mRNA) levels of inflammation markers, indicating a progressive inflammation response, while DEX significantly reversed interleukin-1β (IL-1β) expression at the protein level. Additionally, fibrosis-related markers were markedly increased in the kidneys of UTI mice. These findings were significantly reversed by DEX treatment, revealing its anti-inflammatory and anti-fibrotic activities. DEX reduces the severity of renal fibrosis and inflammation after UTI and exerts a significant renal protective effect.

Novel polyomavirus in the endangered garden dormouse Eliomys quercinus

BackgroundThe garden dormouse (Eliomys quercinus) has experienced a significant population decline across Europe in recent decades. While habitat loss and climate change are often cited as primary factors, pathogen exposure, either to novel or to previously known, may play a role in such a decline. This study aimed to investigate the presence of polyomaviruses in garden dormice, given that these viruses are highly prevalent and can cause disease, particularly in immunocompromised individuals.MethodsThe carcasses of garden dormice (n = 89) were collected throughout Germany. Kidney samples were tested for the presence of polyomavirus DNA using nested degenerate and specific diagnostic PCRs. Seroprevalence was assessed from chest cavity fluid samples through an enzyme-linked immunosorbent assay using polyomavirus VP1 virus-like particles produced in yeast.ResultsA new polyomavirus, related to chimpanzee (Pan troglodytes) polyomaviruses 4 and 5 and human Merkel cell polyomavirus, was identified in the garden dormouse. Two 5,380 bp-length complete viral genomes were sequenced from dormice kidney samples (sequences PQ246041 and PQ246042). Genes encoding the putative structural proteins VP1, VP2, and VP3, as well as the Large, Middle, and small T antigens, containing conserved functional domains were identified. Polyomavirus DNA was detected in 2 of 74 dormice (2.7%, 95% confidence interval: 0-6.4%) through PCR, while 12 of 69 animals (17.4%, 95% confidence interval: 8.4-26.3%) tested positive for polyomavirus-specific antibodies.ConclusionsIn conclusion, here we describe a novel polyomavirus in the garden dormouse with molecular and serological detection. Pairwise sequence comparison and phylogenetic analysis suggest that this novel virus may represent a novel species within the genus Alphapolyomavirus. Future work should examine if this virus is garden dormouse-specific and whether it is associated with disease in dormice.

Insight into distribution and composition of nonhuman N-Glycans in mammalian organs via MALDI-TOF and MALDI-MSI

The major hurdle of xenotransplantation is the immune response triggered by human natural antibodies interacting with carbohydrate antigens on the transplanted animal organ. Specifically, terminal glycoprotein motifs such as galactose-α1,3-galactose (α-Gal) and N-glycolylneuraminic acid (Neu5Gc) are significant obstacles. Little is known about the abundance and compositions of asparagine-linked complex carbohydrates (N-glycans) carrying these motifs in mammalian organs. By studying heart, kidney, and liver tissues from pig, cattle, and sheep, we aimed to gain insights into the abundance and spatial distribution of α-Gal- or Neu5Gc-containing N-glycans. N-glycomes were analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), MALDI-mass spectrometry imaging (MSI), and capillary electrophoresis-electrospray ionization (CE-ESI)-MS. Both α-Gal- and Neu5Gc-containing N-glycans were present in all samples, with α-Gal-modified N-glycans being the most abundant nonhuman carbohydrate motif. The abundance of N-glycans terminating with α-Gal or Neu5Gc was higher in heart and kidney samples than livers. MSI revealed kidneys had the highest glycosylation levels, and α-Gal-containing N-glycans were abundant in the kidney cortex but scarce in the medulla. This study enhances our understanding of α-Gal- and Neu5Gc-modified N-glycans in animal organs and may guide research on carbohydrate antigen-induced immune rejection in xenotransplantation.

Isolation and Molecular Characterization of Pathogenic Leptospira spp. from Brown Rats (Rattus norvegicus) in Zaria and Environs, Kaduna, Nigeria, 2022.

Leptospirosis caused by the pathogenic serovars of Leptospira spp. is a zoonotic disease of global importance. Brown rats (Rattus norvegicus), due to their worldwide distribution and reservoir host status, are considered the most important reservoir for environmental leptospiral persistence and infections in animals. We aimed to isolate, identify, and characterize Leptospira spp. circulating among brown rat populations in Zaria, Kaduna state, Nigeria. Kidney samples from 305 brown rats from different locations in Zaria were collected April-June 2022 and cultured using bacteriologic methods. We also extracted DNA from 24 kidney tissue samples for molecular detection of pathogenic Leptospira spp. and for phylogenetic analysis. Leptospira spp. was detected in 57.7% of the samples by using culture methods and in 91.7% of the samples by using PCR. The phylogenetic tree revealed two distinct clusters among the reference sequences, aligning with the known P1 and P2 subclades. The sequences from this study all clustered together on the phylogenetic tree and were most similar to Leptospira interrogans serovar Pomona strain RZ 11. The high prevalence of Leptospira spp. detected in Zaria suggests that further surveillance and assessment of the public risk are warranted.

Pathological effects and immune modulation in host during Tilapia Parvovirus (TiPV) outbreak in cage and wetland Tilapia farms.

Viral diseases arising in farmed fish are an ongoing challenge to the aquaculture industry, causing severe mortality and economic losses. Recently, there has been a spike in the incidence of a viral disease caused by Tilapia Parvovirus (TiPV) inflicts irreparable damage, and large-scale fish kills in the farmed tilapia species. We investigated a case of disease outbreak and severe mortality in cage and wetland farms of tilapia in West Bengal and Odisha, India. The symptomatic fish showed clinical signs, including hemorrhage, discoloration, ulcer, and redness in the body surfaces. Further analysis revealed that Tilapia Parvovirus was associated (validated by PCR, phylogenetic analysis, and cell line assay) with the infection and mortality of tilapia. The virus was detected in gill, heart, spleen, liver, and kidney samples collected from apparently healthy (asymptomatic) and symptomatic tilapia samples from cage and wetland farms. At the same time, negative results were found in the brain and skin tissue samples. The histological analysis revealed that TiPV induces severe damage invariably in almost all studied tissue, including the liver, kidney, spleen, gill, heart, and brain of tilapia samples. The viral quantification analysis showed that the viral genome was higher in the liver, spleen, and heart than in the tilapia samples' gill, kidney, or brain tissue. Furthermore, the study indicated that TiPV infection has a significant effect on the health of tilapia. The tilapia exhibited an immune reactivity toward TiPV infection (upregulation of chemokine receptors, CRs and interleukin 1β, IL-1β), the majority of the studied immune genes (interleukin 8, IL-8; Toll-like receptors 7, TLR7; tumour necrosis factor α, TNF-α; major histocompatibility complex II, MHC II and nuclear factor kappa B, NF-kB) were significantly downregulated in the kidney, spleen and liver tissue samples of symptomatic tilapia. Further, the in vivo challenge assay confirms that the isolated TiPV is a novel parvovirus pathogen that causes massive mortality in tilapia. The lessons learned from the first cellular and molecular description associated with TiPV epidemiology from wetland and cage farms of tilapia could be critical to developing the current state of the tilapia farming industry. Additionally, a holistic approach is needed to develop management measures to control the virulence and risk factors of TiPV.

Radiographic and tomographic findings of excretory urography of avian kidneys after induced nephropathy in broiler chicken as a model.

Although kidney diseases have a high prevalence, initial recognition and diagnosis of renal disease are complicated in avian medicine. The aim of this study is to assess the intraosseous excretory urography (IOEU) in order to introduce a new method for evaluation of kidney's function in broilers as a primary model of birds. A total of 10 male broiler chickens at the age of 20 days were included and evaluated with plain and serial post-contrast radiography and computed tomography (CT) scan to check the time and concentration of the contrast media (CM) passage from the kidney. In the next step, after the nephropathy induction with gentamicin, plain and serial post-contrast radiography and CT scan were repeated. After the necropsy, kidney samples were sent for histopathology evaluation, and statistical analysis of data obtained from image analysis was conducted. After nephropathy induction, all of the chickens exhibited increase in kidney size and in histopathological evaluation, all the samples (30 of 30) showed severe tubular necrosis and mild glomerulopathy. In the IOEU, the average time to create the maximum concentration of CM, the average time to decrease the CM concentration from the kidney and the time of the complete clearance of CM indicated a significant incensement after nephropathy compared to the time before it. Intraosseous excretory urography (IOEU) is a safe contrast imaging modality and can be useful in the assessment of the urinary system in chickens to check the changes in kidney size and function.

The mercury and low molecular-weight antioxidants levels in ungulates of the Republic of Karelia

The high toxicity of mercury (Hg) poses a danger to the environment and humans, but studies of the concentration of this metal in organisms of terrestrial ecosystems are few. Ecotoxicologists also pay little attention to studying the role of antioxidant vitamins in protecting cells from toxic metals. The Republic of Karelia is one of the northwestern regions of Russia, the biogeochemical features of which can contribute to an increase in the mobility and bioavailability of Hg in food chains. The purpose of the work was to determine the concentration of Hg in the liver, kidneys, muscle and hair of ungulate mammals of the Republic of Karelia (wild boar Sus scrofa L. and moose Alces alces L.) and to analyze the relationship between the level of this toxic metal and the content of low molecular-weight antioxidants – reduced glutathione, retinol and α-tocopherol. Species and tissue-specific of the studied parameters in wild boars and moose are noted. The observations discovered by other researchers that omnivorous species accumulate more Hg in their tissues compared to herbivores, and also that this toxic metal is predominantly accumulated in the kidneys, while muscles contain a minimal amount, have been confirmed. Hg concentrations in most samples of liver and kidney of wild boars and in all samples of these same organs of moose were within the limits recorded for domestic pigs and deer, respectively. The levels of Hg we recorded in the tissues and hair of wild boars and moose were generally comparable to or lower than the levels of this metal noted in animals from other regions of Russia and other countries of the world. In wild boars and moose of Karelia, no statistically significant relationships were found between the Hg level and the content of the studied antioxidants in the internal organs. Moose were characterized by a higher content of α-tocopherol in the body than wild boars, which is a feature of this type of herbivorous ungulate mammal. The results of the study indicate a relatively low level of mercury pollution in terrestrial ecosystems in Karelia.

Vancomycin-induced acute kidney injury in rats: beneficial effects deriving from the co-administration of tadalafil or vitamin C

Background: Vancomycin (VAN) is widely employed in clinical settings for the treatment of severe infections attributable to multi-resistant bacteria, with a notable emphasis on Gram-positive strains. However, its therapeutic usage is significantly hampered by the potential for acute kidney injury (AKI); specifically, the nephrotoxic effects it can exert. In controlled experimental studies, the intraperitoneal (i.p.) administration of VAN at a dose of 400 mg/kg/day over a consecutive seven-day period has been shown to induce considerable AKI in rats. In addressing this concern, vitamin C (VC), recognized traditionally for its antioxidant properties, has been utilized to mitigate various forms of organ damage and inflammatory responses, primarily by diminishing oxidative stress and attenuating inflammation. Concurrently, tadalafil (TAD), a phosphodiesterase type-5 inhibitor commonly associated with the treatment of erectile dysfunction, is also noted for its beneficial anti-inflammatory and antioxidant effects, alongside vasodilatory properties. Aim: This study was designed in order to investigate the protective capabilities of TAD or VC against the VAN-induced nephrotoxic effects in a rat model of AKI. Methodology: A total of 24 male albino Wistar rats were allocated into four distinct groups for the purpose of comparative analysis. Following treatment, both kidney and blood samples were obtained for further evaluation. The study randomized the rats into four categories: group 1 served as the control group, group 2 received VAN at a dosage of 400 mg/kg (via an i.p. injection on a daily basis), group 3 was administered VAN (400 mg/kg; i.p., daily) in conjunction with TAD at a dose of 5 mg/kg daily, and group 4 was given VAN (400 mg/kg; i.p., daily) alongside VC at a dosage of 200 mg/kg daily. Results: Evaluations included the measurement of renal function indicators such as the serum levels of urea, of creatinine, and the creatinine clearance. Additionally, the analysis involved the assessment of antioxidant markers in renal tissue homogenates, including malondialdehyde (MDA), glutathione, and superoxide dismutase. To further elucidate the inflammatory response, indices such as the levels of the tumor necrosis factor-alpha, the interleukins (IL)-1β and IL-6, and the nuclear factor-κB were evaluated within the renal tissue homogenate samples of the studied rats. The administration of VAN resulted in a marked elevation of renal biomarkers and inflammatory indices, alongside a significant reduction in antioxidant levels, with an exception for MDA (which exhibited increased levels); these changes reached statistical significance (p<0.05) when measured against the control group. In contrast, the supplementation with TAD or VC was effective in significantly lowering the serum levels of both renal and inflammatory indicators when compared to the group treated solely with VAN. Furthermore, antioxidant measurements improved, alongside a reduction in MDA levels, thereby demonstrating significant differences (p<0.05). Conclusion: The findings of this study reveal that both TAD and VC provide a degree of renal protection against the VAN-induced damage by enhancing kidney function markers and, concurrently, lessening oxidative stress and inflammatory responses associated with nephrotoxicity.

Sirtuin1 Suppresses Calcium Oxalate Nephropathy via Inhibition of Renal Proximal Tubular Cell Ferroptosis Through PGC-1α-mediated Transcriptional Coactivation.

Calcium oxalate (CaOx) crystals induce renal tubular epithelial cell injury and subsequent nephropathy. However, the underlying mechanisms remain unclear. In the present study, single-cell transcriptome sequencing is performed on kidney samples from mice with CaOx nephrocalcinosis. Renal proximal tubular cells are identified as the most severely damaged cell population and are accompanied by elevated ferroptosis. Further studies demonstrated that sirtuin1 (Sirt1) effectively reduced ferroptosis and CaOx crystal-induced kidney injury in a glutathione peroxidase 4 (GPX4)-dependent manner. Mechanistically, Sirt1 relies on peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α) to promote resistance to ferroptosis in the tubular epithelium, and PGC-1α can recruit nuclear factor erythroid 2-related factor 2 (NRF2) to the promoter region of GPX4 and co-activate GPX4 transcription. This work provides new insight into the mechanism of CaOx crystal-induced kidney injury and identifies Sirt1 and PGC-1α as potential preventative and therapeutic targets for crystal nephropathies.

Toxicological Evaluation of Sargassum polycystum in Mice with Loperamide-Induced Constipation

Constipation gradually increases in people after the age of 50 years leading to difficulty in stool passage. Until now, there have been no effective drugs without side effects. This study investigated the preventive potential and toxicity of Sargassum polycystum extract (SPE) against loperamide-induced constipation in mice. Daily oral administration of SPE (100, 500 and 1000 mg/kg) for 14 days significantly increased defecation frequency in constipation mice. Additionally, no significant differences in blood haematological and biochemical indices between the control, constipation and SPE-treated groups were detected. Histopathological examinations revealed non-toxic effects in liver, kidney and colon tissue samples from the SPE-treated groups compared with the control and constipation groups. Real-time polymerase chain reaction showed that the administration of SPE significantly modulated Saa3, Odc1 and Clu expressions that promote the health of living organisms compared with the control group. Overall, this study showed that daily oral administration of Sargassum polycystumfor 14 days could prevent constipation without any adverse effects. Sargassum polycystumis suitable for further use as a supplement or drug to manage constipation.

Pharmacokinetics and cardioprotective efficacy of intravenous miR-125b* microRNA mimic in a mouse model of acute myocardial infarction.

MicroRNA (miRNA) therapy is a promising approach to induce cardioprotection. We have previously identified cardiac microRNA-125b* (microRNA-125b-2-3p; miR-125b*) as a potential cardioprotective miRNA, termed ProtectomiR. We aimed to characterize the pharmacokinetics and pharmacodynamics, and the effect of miR-125b* mimic on infarct size using an in vivo mouse model. To characterize the pharmacokinetics properties of miR-125b* mimic, a single injection of 10-μg miR-125b* mimic or its scramble miRNA control, or vehicle i.v. was given to C57BL/6 mice. MiR-125b* expression was measured from plasma, heart, kidney and liver samples. Effect of miR-125b* on area at risk and infarct size was assessed after 45-min coronary occlusion, followed by 24-h reperfusion; 10-μg miR-125b* mimic or 10-μg non-targeting miRNA mimic control or vehicle were administered via the right jugular vein at 10th mins of coronary occlusion. To assess molecular mechanism involved in cardioprotection, expression of mRNA targets of miR-125b* were measured from ventricular myocardium at 1, 2, 4, 8 or 24 hpost-treatment using quantitative real time polymerase chain reaction. MiR-125b* expression was markedly increased in plasma and myocardium 1h, and in the liver 2h after treatment. Infarct size was significantly reduced after miR-125b* mimic treatment when compared to the vehicle. The expression of Ccna2, Eef2k and Cacnb2 target mRNAs was significantly reduced 8h after injection of miR-125b* mimic. This is the first demonstration of pharmacokinetic and molecular pharmacodynamic properties as well as the cardioprotective effect of miR-125b* mimic in vivo.

First detection of D181 genotype of infectious bronchitis in poultry flocks of Morocco

BackgroundThis paper reports the first pathological and molecular characterization of the novel variant of infectious bronchitis virus (IBV) D181 in poultry flocks in Morocco and Africa.MethodsThe study includes six poultry farms, involving three flocks of layers aged between 28 and 67 weeks and three broiler flocks aged 27, 39 and 42 days from different regions of Morocco. In all affected layer flocks, a severe drop in egg production with poor eggshell quality was reported. Necropsy of dead birds was carried out, and samples of trachea, lungs, oviduct, ovaries, and kidneys were fixed in 10% neutral buffered formalin for histopathologic examinations, while other portions were stored at -20 °C for molecular analysis. Real time RT-qPCR for IBV gene group was performed, and IBV variants were identified. Partial S1 gene sequences were amplified by conventional RT-PCR, sequenced, and aligned for phylogenetic and amino acid similarity analysis.ResultsNecropsy of dead birds revealed misshapen and hemorrhagic ovarian follicles with an edematous oviduct and severe reaction in the cecal tonsils. A caseous material accumulation in the sinus was noted in few birds. In contrast, the broiler flocks exhibited respiratory clinical signs such as difficulty in breathing, sneezing, tracheal rales, watery eyes and lethargy, associated with a decrease in feed consumption. Mortality in broiler ranged from 2 to 15%. Histopathological analysis of samples showed a lympho-plasmocytic inflammation in the oviduct, trachea, and lungs. Individual necrosis of epithelial cells, with sloughing of the bronchial epithelium and accumulation of desquamated cells with mucus in the airways, was observed in some birds. Partial S1 gene sequencing and phylogenetic analyses showed that the Moroccan strains were very closely related to D181 strains isolated in Dutch layers and breeders in 2018. Nucleotide sequence identities reached 90.9–95% with the Dutch isolates (strain CK/NL/D181/2018).ConclusionOur sequencing results demonstrate for the first time that the D181 IBV genotype is circulating in Moroccan poultry. These findings justify permanent monitoring of circulating strains in order to appropriately adjust vaccination strategies to align with the evolving field situation.

Empagliflozin reduces renal calcium oxalate deposition in hyperoxaluria rats induced with ethylene glycol-ammonium chloride

A retrospective study reported that empagliflozin reduced the risk of urinary stone events in patients with diabetes mellitus. To further investigate empagliflozin's potential, we conducted an animal experiment to determine whether empagliflozin can prevent renal stone formation in hyperoxaluria rats. Hyperoxaluria rat models were constructed by administrating 0.75 % ethylene glycol and 1 % ammonium chloride in water. The empagliflozin-treated rats were gauged with empagliflozin at different concentrations, and their body weight and blood sugar data were recorded. After 30 days of treatment, we obtained 24-h urine, kidney, and blood samples. The urine samples were subjected to component detection. Blood samples were prepared for component detection and cytokines detection. Renal samples were subjected to von Kossa staining, transmission electron microscopy, immunohistochemistry, and transcriptome sequencing analysis. Results showed that in empagliflozin-treated hyperoxaluria rats, renal crystal deposition and mitochondria injury, urinary concentration, and excretion of oxalate were significantly decreased. Additionally, plasma levels of VEGF, IL-2, IL-1β, and MCP-1 were decreased. Immunohistochemistry showed that renal expression of KIM-1, MCP-1 was significantly decreased in empagliflozin-treated hyperoxaluria rats. Transcriptome sequencing of renal tissue represented that 25 genes were down-regulated while 12 were up-regulated in empagliflozin-treated hyperoxaluria rats. These regulated genes were mainly enriched in fatty acid metabolism, insulin resistance, muscle contraction, bile secretion, and parathyroid metabolism. Our animal experiments found that empagliflozin could reduce urinary concentration and excretion of oxalate and inhibit renal inflammation, then abating renal calcium oxalate deposition in hyperoxaluria rats in a non-diabetic state.

Neuropilin-1 as a Key Molecule for Renal Recovery in Lupus Nephritis: Insights from an NZB/W F1 Mouse Model.

Systemic lupus erythematosus (SLE) is an autoimmune disease affecting multiple organs, with lupus nephritis (LN) occurring in 40-50% of SLE patients. Despite advances in diagnosis and treatment, LN remains a major cause of morbidity and mortality, with 10-20% of patients progressing to end-stage renal disease (ESRD). While knowledge of LN's pathogenesis has improved, mechanisms of renal recovery are still largely unexplored. Neuropilin-1 (NRP-1), a transmembrane receptor expressed in renal tissue, has emerged as a promising biomarker for assessing renal recovery in LN. This study evaluates and correlates longitudinal levels of NRP-1 with kidney histology using an NZB/W F1 mouse model of LN. A total of 30 mice were used, with 15 receiving intravenous cyclophosphamide (CYC) and 15 being untreated. NRP-1 levels were measured in urine and serum, and kidney samples were taken from a subgroup of mice for histological evaluation. The results demonstrated a progressive increase in renal and urinary NRP-1 expression, particularly notable at weeks 26 and 32. Urinary NRP-1 levels above 34.40 ng/mL were strong predictors of favorable renal response, showing 100% sensitivity and 88% specificity. These findings indicate a robust correlation between urinary NRP-1 levels and renal histological recovery, underscoring the potential of NRP-1 as a valuable biomarker for assessing renal response in LN. This study demonstrates that renal production of NRP-1 is linked to histological recovery and establishes a foundation for future research into the role of NRP-1 in lupus kidney recovery.

Rapid identification of antibiotic residues in bovine kidney using coated blade spray-mass spectrometry

The use of certain antibiotics in food-producing animals is allowed in Europe following Regulation (EU) 2017/625. However, use could result in antibiotic residues in foodstuffs of animal origin. Maximum residue limits (MRLs) are in place to protect consumers. For monitoring purposes, animal matrices are tested to verify their compliance with these MRLs. Initially, matrices of (slaughtered) food animals are screened, often using a microbiological assay. Faster screening tests for antibiotics would be an advantage for control laboratories. Therefore, the present study describes, for the first time, the use of coated blade spray (CBS) followed by direct mass spectrometry (MS) analysis for the screening of tetracyclines, sulfonamides, quinolones, and macrolides residues from the renal area of intact bovine kidneys. An optimized workflow using two different desorption/ionization solutions per blade allowed screening of target compounds within 1 min per sample. The proof-of-principle of the CBS-MS method is validated according to (EU) 2021/808, presenting CCβ screening values of 0.1 × MRL for 43 analytes, 0.5 × MRL for 4 analytes, and 2.5 µg kg−1 for the prohibited substance dapsone, respectively. The developed method was successfully applied to seven official control samples of bovine kidneys. One of these samples was found to be positive using the CBS-MS method, which was confirmed as a true positive by LC-MSMS analysis. The developed method demonstrates that CBS devices can directly extract and analyze kidney samples for food safety testing.

Simultaneous determination of macrolide and quinolone antibiotics in animal tissues for human consumption validated by Regulation (EU) 2021/808

In this work, an analytical method for the simultaneous detection and quantification of 11 macrolides (erythromycin A, gamithromycin, lincomycin, neospiramycin, pirlimycin, spiramycin, tildipirosin, tilmicosin, tulathromycin A and its metabolite CP-60300, and tylosin A) and 10 quinolones (ciprofloxacin, danofloxacin, difloxacin, enrofloxacin, flumequine, marbofloxacin, nalidixic acid, norfloxacin, oxolinic acid, and sarafloxacin) in muscle (bovine, ovine, porcine, equine, rabbit, poultry including laying hens, and fish) and kidney (bovine, ovine, porcine, equine) for human consumption was developed and validated according to Commission Implementing Regulation (EU) 2021/808. Sample preparation consisted of liquid extraction using a citrate–phosphate/EDTA buffer solution (McIlvaine buffer) followed by solid phase extraction for cleanup and UHPLC-MS/MS analysis. The developed method was comprehensively validated with regard to relative matrix effects (CVMF ≤ 20 %), linearity (R2 ≥ 0.98), accuracy (84–119 %), trueness (−16 to + 19 %), repeatability (2.1–20.0 %), reproducibility (3.0–25.0 %), selectivity, and robustness. The method limit of quantification values ranged from 2 to 40 µg kg−1, established as 0.1 × the lowest maximum residue limit (MRL) by species for the authorized antibiotics. Method applicability was demonstrated in six interlaboratory proficiency tests, and in the official control analyses of 1200 field samples of muscle and kidney.

The integrated effect of roflumilast and selenium nanoparticles on nephrotoxicity generated by cisplatin through the regulation of the antioxidant and apoptotic pathways

AimThe current investigations aimed to investigate the potential synergistic effect of Roflumilast (ROF) and Selenium nanoparticles (SeNPs) administration on Cisplatin (Cis) -induced nephrotoxicity. Materials and methodsFifty male rats were divided into five groups; Control group: animals were administered 0.9 % saline solution. Cis group: animals were injected with a single dose of 6 mg/kg. ROF group: Rats received a dosage of 1.2 mg/kg orally daily for 11 days. SeNPs group: animals orally received 0.5 mg/kg of ROF daily for 11 days. The ROF + SeNPs group was administered both after receiving a Cis injection for 11 days. Animals were sacrificed at 5 and 11 days, and the urine and blood samples were collected on day 5 and day 11 for chemical analysis, while kidney samples were obtained for molecular, histological, and immunohistochemical studies. ResultsThe levels of serum creatinine, Blood urea nitrogen (BUN), and total protein were elevated in the Cis group compared to the control group (p < 0.05). While the combination of ROF and SeNPs dramatically decreased these values after 5 and 11 days (p < 0.05). In addition, Cis caused renal oxidative stress by elevating MDA levels and suppressing the activities of SOD, GSH, and CAT. Similarly, these effects were modulated by ROF and SeNPs after 11 days (p < 0.05). Furthermore, the concurrent administration of ROF and SeNPs resulted in a significant increase in the expression of HO-1, Nrf2, and Bcl2, while decreasing the expression of BAX and IL-6 compared to the Cis group after 11 days (P < 0.05). ConclusionThe study showed that both ROF and SeNPs had significant therapeutic potential in reducing the pathological alterations caused by Cis.