Sections Of Renal Cortex Research Articles

O237: Conversion of human kidney tissue to Bombay type using FucOB enzyme

Abstract Introduction The Bombay phenotype (Oh) is characterised by an absence of H antigen expression on red blood cells and vascular endothelium. The Oh phenotype is rare but most prevalent in the Indian subcontinent where it poses significant challenges in performing blood transfusions and kidney transplants, as Oh recipients can only receive from Oh blood type donors. The enzyme α-1,2-fucosidase (FucOB) derived from Akkermansia muciniphila bacteria has been shown to hydrolyse blood group H antigens. This study examined the efficiency of the FucOB enzyme on O type human kidney tissue to obtain Bombay phenotype. Methods H antigen removal was investigated in formalin-fixed paraffin embedded (FFPE) human kidney cortical biopsies from blood group O-type donors (n=8). Biopsies were incubated at 37OC for 1h with FucOB enzyme at concentrations 0µg/ml, 0.05µg/ml, 0.1µg/ml, 0.5µg/ml and 5µg/ml and inactive enzyme (iFucOB; 5µg/ml). Quantitative immunofluorescence was used to determine co-localisation of H antigen and vascular endothelium (Ulex). Results Increasing concentrations of FucOB progressively removed H antigen from FFPE sections of renal cortex. When compared to untreated control and iFucOB, a FucOB concentration of 5µg/ml caused 95% loss of H antigen expression on renal vascular endothelium (p<0.0001; Friedman test with Dunn’s multiple comparisons test). Conclusion This is the first description of enzymatic cleavage of H antigen from human O-type kidney tissue. The next step is to deliver FucOB enzyme to human kidneys during normothermic machine perfusion. This strategy has the potential to dramatically increase the availability of ABO compatible kidneys from individuals with Bombay phenotype.

Mitigative Effects of Alpha-lipoic Acid on the Toxicity of Dimethoate in Male Rats

Background and Aim: Organophosphates are currently widely used by humans, but these compounds have tremendous negative effects on human health. Therefore, this study aims to use alpha-lipoic acid (ALA) to alleviate the negative effects of dimethoate (DM). Methods: This study was designed as follows: Thirty adult male Wistar albino rats were utilized and further subdivided into control, DM and DM+ALA groups. Liver and renal cortex sections from all groups were processed for histopathological examination and biochemical estimation of liver function tests, serum urea, creatinine, BUN, testosterone and lipid profile was performed. Results: This study clarified the ameliorative effects of ALA on the negative effects of DM, where ALA induced a significant recovery of hepatic (ALT, AST, ALP and total protein) and renal functions by normalizing them in the DM + ALA group and promoted some improvement of lipid profile and testosterone levels. Additionally, ALA restored normal hepatic and renal histomorphology. Conclusion: ALA therapy can be concluded to ameliorate the negative effects of DM that affect vital organs such as the liver and kidney. Additionally, ALA can reduce the occurrence of atherogenesis by decreasing the levels of unhealthy cholesterol in the blood. ALA also boosts testosterone levels, thus augmenting male sexual characteristics.

Aqueous garlic extract supresses experimental gentamicin induced renal pathophysiology mediated by oxidative stress, inflammation and Kim-1

BackgroundGentamicin (Gent) has rapid & high bactericidal action in addition to its cheap price. Nevertheless, 30% of gentamicin-treated patients develop nephrotoxicity. ObjectiveTo explore the probable nephroprotective effects of the aqueous garlic extract (AGE) & to elucidate its underlying mechanisms via monitoring proinflammatory cytokines as tumer necrosis factor (TNF-α), interleukin 6 (IL-6) and interferon-γ (INF-γ), oxidative stess markers as malondialdehyde (MDA) & superoxide dismutase (SOD) & kidney injury molecule (Kim-1) as a promising early specific biomarker of renal dysfunction. Methods32 adult male rats were divided into 4 equal groups treated for 21 days as: normal control group received normal saline orally, AGE-treated group received AGE at 250 mg/kg/day orally, Gent-treated group received Gent-sulphate intraperitoneal injection at 80 mg/kg /day, and AGE & Gent cotreated group received AGE and Gent concomitantly in the same previous doses. Serum urea, creatinine, glomerular filteration rate (GFR), systolic (SBP) and diastolic blood pressure (DBP), TNF-α, IL-6, INF-γ, MDA and SOD and Kim-1 mRNA expression were evaluated in kidney tissue homogenate. Renal cortex sections stained with Haematoxylin & eosin (H&E) were examined. ResultsAGE is nephroprotective through significantly reducing serum urea, creatinine, SBP and DBP, TNF-α, IL-6, INF-γ and MDA (the main product of lipid peroxidation), decreasing expression of Kim-1 mRNA in renal tissue and increasing level of GFR, the natural antioxidant SOD and improving renal histological features of Gent-treated rats. ConclusionAGE normalizes Gent-induced renal dysfunction. Their co-administration is a plausible advice, although the therapeutic efficiency of Gent was not investigated.

Synergistic therapeutic effects of Vitis vinifera extract and Silymarin on experimentally induced cardiorenal injury: The pertinent role of Nrf2

BackgroundCardiorenal crosstalk has gained growing scientific curiosity recently. Clinical observations have approved that heart and kidney performances are intimately interrelated; acute or chronic dysfunction of either is inevitably mirrored on the other. This coexistence usually has the poor prognosis and worsened outcome. MethodsWe designed this study to explore therapeutic potentials of combined Vitis vinifera and Silymarin extracts on histopathological alterations of experimentally induced cardiorenal injury model. Moreover, to examine the pertinent role of Nrf2 in their bio-molecular actions. Sixty adult male Wistar albino rats were utilized, further subdivided into control, doxorubicin (DXR), DXR + Silymarin, DXR + Aqueous Vitis, DXR + Ethanolic Vitis, DXR + Ethanolic Vitis + Silymarin. Left ventricle and renal cortex sections from all groups were processed for histopathological examination, biochemical estimation of serum Urea, Creatinine, BUN, lipid profile and hs-CRP and real-time PCR of Nrf2 expression in cardiac and renal tissue homogenate were performed. ResultsOur results proved that combined ethanolic extract of Vitis vinifera and Silymarin restored normal renal and cardiac histomorphology. Significant improvement of Creatinine, BUN, lipid profile and hs-CRP cardiac and renal biochemical indicators confirmed our results. Moreover, significant elevation of mRNA expression levels of Nrf2 proved that combined Vitis vinifera and Silymarin action was directly related to the redox-sensitive regulator pathway. ConclusionsWe concluded that synergistic therapeutic effect of Vitis vinifera extract and Silymarin on experimental cardiorenal injury model owes principally to promoting activation of the Keap1/Nrf2 signaling pathway.

ACE INHIBITORS VS. AT1 RECEPTOR BLOCKERS: RENAL HEMODYNAMICS IN AN EXPERIMENTAL MODEL OF CONGESTIVE HEART FAILURE

Gentamicin (Gent) has rapid & high bactericidal action in addition to its cheap price. Nevertheless, 30% of gentamicin-treated patients develop nephrotoxicity.To explore the probable nephroprotective effects of the aqueous garlic extract (AGE) & to elucidate its underlying mechanisms via monitoring proinflammatory cytokines as tumer necrosis factor (TNF-α), interleukin 6 (IL-6) and interferon-γ (INF-γ), oxidative stess markers as malondialdehyde (MDA) & superoxide dismutase (SOD) & kidney injury molecule (Kim-1) as a promising early specific biomarker of renal dysfunction.32 adult male rats were divided into 4 equal groups treated for 21 days as: normal control group received normal saline orally, AGE-treated group received AGE at 250 mg/kg/day orally, Gent-treated group received Gent-sulphate intraperitoneal injection at 80 mg/kg /day, and AGE & Gent cotreated group received AGE and Gent concomitantly in the same previous doses. Serum urea, creatinine, glomerular filteration rate (GFR), systolic (SBP) and diastolic blood pressure (DBP), TNF-α, IL-6, INF-γ, MDA and SOD and Kim-1 mRNA expression were evaluated in kidney tissue homogenate. Renal cortex sections stained with Haematoxylin & eosin (H&E) were examined.AGE is nephroprotective through significantly reducing serum urea, creatinine, SBP and DBP, TNF-α, IL-6, INF-γ and MDA (the main product of lipid peroxidation), decreasing expression of Kim-1 mRNA in renal tissue and increasing level of GFR, the natural antioxidant SOD and improving renal histological features of Gent-treated rats.AGE normalizes Gent-induced renal dysfunction. Their co-administration is a plausible advice, although the therapeutic efficiency of Gent was not investigated.

Glomerulosclerosis in transgenic rabbits with ubiquitous Venus protein expression.

Focal segmental glomerulosclerosis (FSGS) is a potential cause of nephrotic syndrome both in humans and pet mammals. Glomerulopathy was reported earlier in green fluorescent protein (GFP) transgenic (TG) mice, but glomerulosclerosis has not been examined in GFP TG rabbits so far. In the present study, the potential manifestation of FSGS was investigated in both Venus TG rabbits generated by Sleeping Beauty (SB) transposition and age-matched control New Zealand White (NZW) rabbits. Venus protein fluorescence was detected by confocal microscopy and quantified by microplate reader. Urinalysis, haematology, serum biochemistry and renal histology were performed to assess the signs of FSGS. Higher levels of Venus fluorescence were determined in renal cortex samples than in the myocardium by both methods. Urinalysis revealed proteinuria in Venus heterozygote TG bucks, while Venus homozygote TG bucks developed microscopic haematuria. Supporting the urinalysis data, the histological findings of FSGS (glomerulomegaly and sclerotic glomeruli) were observed in renal cortex sections of Venus TG rabbits. Taken together, Venus TG bucks were diagnosed with FSGS; thus, this type of glomerulopathy could be a common disease in TG animals overexpressing GFP.

Effect of osteopontin knockdown in the kidneys of hyperoxaluric rats on renal calcium oxalate crystal deposition

Objective To address the effects of osteopontin (OPN) on calcium oxalate renal calculus formation. Methods We silenced the OPN gene in rat kidney in vivo using lentivirus vector-deliverd short hairpin RNA (shRNA) targeting OPN through renal venous transduction. On the day 7 after injection with lentivirus, rats were fed on water containing 0.8% glycol and 1% ammonium chloride for 9 days. At the end of the experiments, the rats were killed and the kidney tissues were removed for subsequent analysis. Results The lentivirus vector could effectively infect renal cortex and medulla tissue. OPN-shRNA transfection resulted in significant reduction in the content of OPN. Crystals recorded as the average total pixels (object area) were 20 716±7 755 and 54 921±3 469 in renal cortex sections in the OPN-shRNA group and negative lentivirus vector group, respectively (P<0.01), as well as 18 618±2 285 and 30 475±1 735 in the medulla sections in both groups, respectively (P<0.01). Conclusion Specific knockdown of OPN gene in kidneys of hyperoxaluric rats leads to a decrease in OPN production and significantly inhabits calcium oxalate crystal deposition. OPN may play a certain role in promoting calcium oxalate renal calculus formation. Key words: Osteopontin; Calcium oxalate; Nephrolithiasis; RNA interference

Cloisonné kidney in mouflon sheep ( Ovis orientalis musimon )

This study describes a case of kidney associated with infection by Babesia ovis in a wild mouflon ( Ovis orientalis musimon j found dead in year in eastern Sierra Morena (Andalusia, Spain). The major macroscopic lesions were a) icteric coloration in ocular and oral mucosa, and b) dark metal coloration on the kidneys' external surface. Intense dark coloration alternating with intense greyish strips was observed on renal cortex sections. Blood samples were collected for haematological and parasitological analyses, as well as samples from various organs for histopathological examination. Infection by B. ovis was confirmed by using the Polymerase Chain Reaction test (PCR). Histopathological examination revealed tubulonephrosis associated with thickening of the basal membrane of the convoluted portions of the renal tubules (both proximal and distal), which took on an intense brownish pigmentation. These renal lesions have been described as cloisonne kidney. Cloisonne kidneys show uncommon coloring of the renal tubular system, associated with ferritin and hemosiderin deposits in these structures resulting from a chronic hemolitic process (hemoparasites, toxins, etc.). In our case, hemolitic phenomena are associated with infection by B. ovis . The seroprevalence of this hemoparasite in wild mouflon populations in other regions of Spain (Catalonia) is up to 15%.

Experimental study on AT1-receptor-peptide-induced myocardial immune damage in rat.

In order to investigate the immunological damage in rat immunized with AT1-receptor peptide, 18 male Wistar rats were divided into two groups: immunized-group (n = 12), each rat was immunized with 150 micrograms AT1-receptor peptide coupled to bovine serum albumin, together with Freund's adjuvant. Control group (n = 6), sham-immunized, "immunized liquid" was same as immunized-group except AT1-receptor peptide. Systolic blood pressure (SBP) was measured by using the tail-cuff technique, antibody against AT1-receptor peptide detected by using ELISA method, and left ventricular myocardium and renal cortex sections were observed under light and electron microscopy. There was no significant difference in SBP and light microscopic observation of the tissue sections between the immunized-group and control group. The O.D. value of anti-AT1-receptor peptide antiserum was significantly higher in the immunized-group than in the rats before immunization and control group (P < 0.01). Positive rate in the immunized-group was 100%, while 0% in the control group. Ultramicroscopic morphology showed potential myocardial injury, including: increase in number of mitochondria, swelling of many mitochondria with reduction in number or absence of their cristae and cristolysis, disorder of the cardiac myofibrils, and myofibrillar disruption and myocytolysis. And lysosomes were increased in renal tubular epithelia. The AT1-receptor peptide could induce to generate the antibody against AT1-receptor peptide and lead to myocardial and renal damage in rats.

Recent declines of hypertensive renovasculopathies in New Orleans blacks

Some evidence indicates that renovasculopathies measured in paraffin sections of renal cortex obtained at autopsy might offer a general guide to prevailing mean blood pressure (MBP) in the averages of demographic groups. In this study, data on forensic autopsies done in New Orleans from 1968 to 1998 were retrieved from computer archives. Observations are confined to basal cases (ie, those with causes of death having no known correlation with hypertension and therefore, who can be taken as approximately representative of the sampled populations, although overt hypertensives may be underrepresented in such a series). MBP derived from renovasculopathies were used to follow birth date cohorts born around the years 1928, 1938, 1948, 1958, 1968, and 1978. Vasculopathy-derived MBP of white men and women increased with age in a stable pattern with no shifting of positions among the cohorts. This pattern offered a baseline for comparison with the data on blacks. Against this baseline, blacks displayed the highest MBP in the earliest cohort with stair-step declines in later born cohorts, and an abrupt decrease between the 1958 and 1968 cohorts. Data from National Health And Nutrition Examination Surveys are generally supportive of these conclusions, but are of uncertain significance because of severe downward drift due to method changes in the National Health And Nutrition Examination Survey III 1988–91 survey. The data analyzed here provide no direct evidence on the matter of whether we are gaining better blood pressure control in hypertensive individuals who really need it.

Pharmacological characterisation and autoradiographic localisation of a putative dopamine D 3 receptor in the rat kidney

The pharmacological profile and the microanatomical localisation of a putative dopamine D 3 receptor in the rat renal cortex were investigated using radioligand binding assay and light microscope autoradiography techniques. [ 3 H ]7-hydroxy- N, N-di- n-propyl-2-aminotetraline ([ 3 H ]7-OH-DPAT) was used as a ligand. [ 3 H ]7-OH-DPAT was bound specifically to sections of renal cortex. The binding was time-, temperature-, concentration-dependent, of high affinity and guanine nucleotide-insensitive. The dissociation constant ( K d) value was 0.57±0.02 nM and the maximum density of binding sites ( B max) was 62.4±3.5 fmol/mg tissue. The pharmacological profile of [ 3 H ]-7-OH-DPAT binding to sections of rat renal cortex suggests the labelling of a dopamine D 3 receptor. Light microscope autoradiography revealed the accumulation of the radioligand primarily within cortical tubules and to a lesser extent in the glomerular tuft. In glomeruli, binding sites were found mainly in mesangium and mesangial cells. The demonstration of a putative dopamine D 3 receptor in slide-mounted sections of rat renal cortex suggests that appropriate radioligand binding assay techniques combined with autoradiography, may contribute to characterise peripheral dopamine receptor subtypes.

Expression and tissue localization of donor‐specific complement C3 synthesized in human renal allografts

Recent evidence suggests that the third component of complement, C3, is synthesized in renal tissue, and that increased C3 synthesis occurs in allograft rejection and immune complex-mediated nephritis. However, it is unclear whether intrinsic renal cells or migratory cells in the inflammatory infiltrate, possibly of recipient bone marrow origin, are the source of the C3 detected. This was investigated by determining the C3 allotypes of mRNA and protein produced by transplanted human kidney. Twenty donor-recipient pairs were examined, of which nine pairs had C3 allotypes that were informatively mismatched at the C3 F/S locus. Reverse transcriptase polymerase chain reaction (RT-PCR) followed by amplification refractory mutation system analysis showed intracellular donor-specific mRNA expression in six of these nine cases, at up to 61 days post-transplantation. Nested PCR reactions and the size of PCR products excluded contamination by genomic DNA. Allotype-specific staining of frozen sections of renal cortex demonstrated donor-derived C3 protein in both glomeruli and tubules of all biopsies examined, in a predominantly tubular distribution. These results imply that at least some of the pro-inflammatory effects of complement arise from intrinsic tissue synthesis of donor C3, and that this may represent a previously unrecognized source of tissue injury. The occurrence of local synthesis of C3 of donor allotype may have functional implications related to C3 allotype, and may also be relevant to strategies to inhibit intrarenal complement-mediated injury.

Detection of antibodies against mitochondria by the fluorescent antibody technic with smears of rat liver cells.

SERA of patients with primary biliary cirrhosis contain autoantibodies against a non-organ specific cytoplasmic component, which are probably mitochondria1,2. A convenient way to detect these antibodies is by the fluorescent antibody technique using unfixed cryostat sections of human or rat organs. Sections of renal cortex proved to be the most suitable for the demonstration of cytoplasmic antibodies. Liver sections showed the same coarsely granular staining, but because of the uniform appearance of this tissue, it was difficult to distinguish the fluorescence obtained with the weaker sera from non-specific artefacts1.