Extrarenal Tissues Research Articles

Differential Effect of Aldosterone or Mineralocorticoid Receptor Overexpression on Retinal Inflammation.

Overactivation of the mineralocorticoid receptor (MR) pathway is proinflammatory and contributes to the pathogenesis of diabetic retinopathy and of age-related macular degeneration. Excess of aldosterone, the specific MR ligand, is known to stimulate the production of proinflammatory cytokines and chemokines in extrarenal tissues and cells. In the RPE/choroid complex, aldosterone upregulated genes encoding proteins of the inflammatory response and downregulated genes encoding proteins involved in synaptic activity and neurotransmitters. Yet, cortisol, which is the main MR ligand in the eye, is a potent anti-inflammatory endogenous glucocorticoid. The aim of the present work was to better understand the role of MR activation in retinal inflammation either by acute injection of aldosterone or overexpression of the receptor. We first analyzed the retinal transcriptomic regulation induced by acute intraocular injection of aldosterone in the rat. Then, we used a transgenic rat overexpressing human MR (hMR) to also conduct retinal transcriptomic analysis as well as histological evaluation of the retina, retinal pigment epithelium and choroid. Our results show that acute intravitreal injection of aldosterone is highly proinflammatory, upregulating pathways related to microglial activation, oxidative stress, cell death, and downregulating pathways related to glial/neuronal cells activity and proper neurotransmission. On the other hand, hMR overexpression mediates a low-grade inflammation in the retina, associated with notable choroidal inflammation and choroidal neuropathy. Consequences of hMR overexpression or aldosterone-injection on retinal transcriptome reveal very distinct pathological mechanisms, with only a few common genes regulated, most of them not being regulated in the same way. Although aldosterone is highly proinflammatory in the retina, MR overactivation in its physiologic milieu mediates a low-grade inflammation in the neural retina.

An efficient inducible model for the control of gene expression in renin cells.

Fate mapping and genetic manipulation of renin cells have relied on either noninducible Cre lines that can introduce the developmental effects of gene deletion or bacterial artificial chromosome transgene-based inducible models that may be prone to spurious and/or ectopic gene expression. To circumvent these problems, we generated an inducible mouse model in which CreERT2 is under the control of the endogenous Akr1b7 gene, an independent marker of renin cells that is expressed in a few extrarenal tissues. We confirmed the proper expression of Cre using Akr1b7CreERT2/+;R26RmTmG/+ mice in which Akr1b7+/renin+ cells become green fluorescent protein (GFP)+ upon tamoxifen administration. In embryos and neonates, GFP was found in juxtaglomerular cells, along the arterioles, and in the mesangium, and in adults, GFP was present mainly in juxtaglomerular cells. In mice treated with captopril and a low-salt diet to induce recruitment of renin cells, GFP extended along the afferent arterioles and in the mesangium. We generated Akr1b7CreERT2/+;Ren1cFl/-;R26RmTmG/+ mice to conditionally delete renin in adult mice and found a marked reduction in kidney renin mRNA and protein and mean arterial pressure in mutant animals. When subjected to a homeostatic threat, mutant mice were unable to recruit renin+ cells. Most importantly, these mice developed concentric vascular hypertrophy ruling out potential developmental effects on the vasculature due to the lack of renin. We conclude that Akr1b7CreERT2 mice constitute an excellent model for the fate mapping of renin cells and for the spatial and temporal control of gene expression in renin cells.NEW & NOTEWORTHY Fate mapping and genetic manipulation are important tools to study the identity of renin cells. Here, we report on a novel Cre mouse model, Akr1b7CreERT2, for the spatial and temporal regulation of gene expression in renin cells. Cre is properly expressed in renin cells during development and in the adult under basal conditions and under physiological stress. Moreover, renin can be efficiently deleted in the adult, leading to the development of concentric vascular hypertrophy.

Interplay between caveolin-1 and mineralocorticoid receptor in cardiometabolic disease.

Over the past decades, research has clearly established the important role of the mineralocorticoid receptor (MR) in both renal and extra-renal tissues. Recently, caveolin-1 (Cav-1) has emerged as a mediator of MR signaling in several tissues, with implications on cardiovascular and metabolic dysfunction. The main structural component of caveolae (plasma membrane invaginations with diverse functions), Cav-1 is a modulator of cardiovascular function, cellular glucose, and lipid homeostasis, via its effects on signal transduction pathways that mediate inflammatory responses and oxidative stress. In this review, we present evidence indicating an overlap between the roles of the MR and Cav-1 in cardiometabolic disease and the relevant signaling pathways involved. Furthermore, we discuss the potential use of Cav-1 as a biomarker and/or target for MR-mediated dysfunction.

A Rare Synchronous Existence of Warthin’s Tumour and Oral Cancer

Oral squamous cell carcinoma (OSCC) is one of the most common malignancies worldwide while Warthin’s tumor (WT) is a common type of benign salivary gland neoplasm mostly seen in the parotid gland but rarely involves extra parotid tissues. Extra parotid WT is itself a rare entity and its synchronous occurrence with OSCC is an unusual phenomenon. It is vital to accurately identify the pathology as confusion may arise due to its rare occurrence and an intriguing presentation of resembling metastasis in lymph nodes. In the present review, a systematic literature search was performed and case reports and series in which synchronous existence of WT and OSCC were included. Total 13 papers have been included with 17 cases of synchronous WT and OSCC. Quality assessment for cases was done based on the CARE guidelines. The given results describe the histological features and site of WT in individuals who also had OSCC. Most of the studies described the histological findings of WT, which were seen involving the lymph nodes. More than 50% of the included cases had a history of tobacco chewing or smoking. The cases that have reported for follow up showed no evidence of any recurrence. WT with OSCC synchronous occurrence at a different site from the primary tumor can mimic a metastatic lesion from a primary lesion. It is a diagnostic challenge and could alter the management of these patients if not identified accurately. Awareness about this synchronous occurrence can avoid the overtreatment in such cases. Majority of the WTs described were seen in cervical lymph nodes.
 Keywords: Oral cancer; Warthin tumor; Benign Neoplasm; Mixed salivary gland tumor; Synchronous neoplasm; metastasis; Second malignancy; Prognosis.

Impaired systemic proteostasis and peripheral immune cell dysfunction in kidney diseases

ABSTRACT Kidney diseases, encompassing a spectrum of either acute or chronic disorders, manifest complex systemic repercussions beyond renal dysfunction. These include a reduced efficacy of tissue proteostasis mechanisms to control protein synthesis, folding, and degradation in both the renal and extra-renal compartments. Emerging research has revealed a pivotal interplay between proteostasis and immune regulation in the pathogenesis of kidney diseases, along with their cardiovascular, immunometabolic, and organ dysfunction symptoms. This review outlines the multifaceted connections between proteostasis dysregulation and immune dysfunction in kidney disease onset and progression. The crosstalk between the proteostasis network and immune cells orchestrates a bidirectional communication that amplifies pathological cascades, exacerbating kidney injury and impairing organ resilience. Furthermore, the article focuses on the implications of systemic proteostasis defects in modulating immune responses in extra-renal tissues underscore the systemic nature of kidney diseases. The disruption of proteostasis mechanisms triggers the release of damage-associated molecular patterns (DAMPs) and inflammatory mediators, perpetuating a state of heightened immune activation, contributing to systemic complications in affected individuals. Understanding the intricate interaction between proteostasis and immune regulation in kidney diseases, including both chronic and acute forms, promises novel therapeutic interventions. Targeting proteostasis pathways to restore cellular homeostasis and modulating immune responses could offer innovative strategies to mitigate renal damage and ameliorate systemic complications associated with kidney diseases. Harnessing this knowledge may also pave the way for the development of more efficient dialysis therapies and interventions with sorbents and hemoperfusion methods, to improve the clinical outcome of the patients.

Sweet taste receptors play roles in artificial sweetener-induced enhanced urine output in mice

Sweet taste receptors found in oral and extra oral tissues play important roles in the regulation of many physiological functions. Studies have shown that urine volume increases during the lifetime exposure to artificial sweeteners. However, the detailed molecular mechanism and the general effects of different artificial sweeteners exposure on urine volume remain unclear. In this study, we investigated the relationship between urinary excretion and the sweet taste receptor expression in mice after three artificial sweeteners exposure in a higher or lower concentration via animal behavioral studies, western blotting, and real-time quantitative PCR experiment in rodent model. Our results showed that high dose of acesulfame potassium and saccharin can significantly enhance the urine output and there was a positive correlation between K+ and urination volume. The acesulfame potassium administration assay of T1R3 knockout mice showed that artificial sweeteners may affect the urine output directly through the sweet taste signaling pathway. The expression of T1R3 encoding gene can be up-regulated specifically in bladder but not in kidney or other organs we tested. Through our study, the sweet taste receptors, distributing in many tissues as bladder, were indicated to function in the enhanced urine output. Different effects of long-term exposure to the three artificial sweeteners were shown and acesulfame potassium increased urine output even at a very low concentration.

A selective mineralocorticoid receptor blocker, esaxerenone, attenuates vascular dysfunction and inhibits atherogenesis in apolipoprotein e-deficient mice

Abstract Introduction Pharmacological blockade of mineralocorticoid receptors (MR) is a potential therapeutic approach to reduce cardiovascular complications because MR play a crucial role in cardiovascular regulation. Recent studies suggest that MR antagonists affect several extrarenal tissues, including vascular function. We investigated the effects of a novel non-steroidal selective MR blocker, esaxerenone (ESAX), recently approved for hypertension treatment in Japan, on vascular function and the development of atherosclerosis. Methods and results ESAX (3mg/kg/day) or vehicle was orally administered to western type diet-fed apolipoprotein E-deficient mice for 20 weeks or for 8 weeks to investigate the effects on atherogenesis or vascular dysfunction. Administration of ESAX for 20 weeks suppressed the development of atherosclerotic plaques in the aortic arch compared with the vehicle group (p<0.001) without alteration of blood pressure. In addition, ESAX reduced the expression of intercellular cell adhesion molecule-1 (p<0.05), macrophage infiltration (p<0.01), lipid deposition (p<0.05) and tended to decrease the expression of vascular cell adhesion molecule-1 as determined by immunohistochemical analysis. Moreover, phosphorylation of eNOSSer1177 (p<0.05) and Akt (p<0.05) were increased in the aorta of ESAX-treated mice. Administration of ESAX for 8 weeks attenuated western type diet-induced endothelial dysfunction (p<0.05) accompanied with the decrease in expression of ICAM1 (p<0.05) and phosphorylation of JNK (p<0.05) in the descending aorta. In in vitro experiment, aldosterone decreased the phosphorylation of eNOSSer1177 and increased the phosphorylation of eNOSThr495 in a dose dependent manner, which were attenuated by ESAX in HUVECs. In ex vivo vascular reactivity assay, ESAX diminished the aldosterone-induced endothelial dysfunction. Conclusion ESAX ameliorated hyperlipidemia-induced atherosclerosis and vascular dysfunction in mice, suggesting that ESAX is a promising therapeutic approach for cardiovascular complications.

SPINAL TUMORS-05 CERVICAL DENDRITIC FOLLICULAR SARCOMA: ABOUT A CASE WITH LITERATURE REVIEW

Abstract Dendritic follicular sarcoma (DFS) is a very rare and often poorly known neoplasia. It develops mainly in peripheral lymph nodes at the expense of dendritic follicular cells. However, extra nodal locations were also found in the literature. Diagnosis is often difficult, based mainly on histopathological and immunohistochemical data. There is currently no consensual therapeutic strategy proposed by the National Understanding Cancer Network. The aim of this work is to focus on the various epidemiological, histopathological, immunohistochemical and therapeutic aspects of the follicular dendritic sarcoma. We report the case of a cervical dendritic follicular sarcoma with literature review. This is a 51-year-old patient with no particular pathological history, consulting for a right lateral cervical mass evolving for 5 years without dyspnea, dysphagia, dysphonia, nor other associated signs evolving in a context of apyrexia and general state conservation. The clinical examination had objectified a firm, fixed, painless right upper jugulocarotid mass without inflammatory signs in sight. The cervical MRI had objectified two contigüe lateral cervical tissue masses well encapsulated, well limited necrotic with mass effect on the larynx without vascular enclosing. The patient had benefited from tumor reduction. Histology was in favor of a dendritic follicular sarcoma. Cervical CT was performed 2 months postoperatively which had shown a mixed right lateral cervical mass with thickened wall and hypodense center pushing in and back the primary carotid and forward the internal jugular vein without any sign of invasion locoregional. The patient then benefited from a complete removal of the tumor mass with ligation of the internal jugular vein. The anatomopathological study had objectified a tumor proliferation made of fusiform cells with nuclei augmented of irregular volume, nucleoli presenting moderate atypies without peri nervous engainement or neoplastic vascular emboli. Immunohistochemistry had shown positive antibodies anti-CD23, anti-CD35, anti-vimentin and anti-PS100. The Ki-67 index was positive on 15% of the cells. This profile concluded in a dendritic follicular sarcoma. The post-operative follow-up was simple. The patient was then referred to oncology for complementary radiation therapy. At 6 months no sign of recurrence was noted. Dendritic follicular sarcoma (DFS) is a relatively rare entity derived from dendritic follicular cells, which are B-cell antigen-presenting cells in the germinal centres of lymphoid follicles. It mainly affects the peripheral lymph nodes especially those of the head and neck. However, extra nodal tissues are affected in about 30% of cases especially the pharynx, oral cavity, thyroid gland. The diagnosis of SFD is difficult and often poorly understood, due to its silent symptomatology, extremely varied morphology and aspecific imaging. The main differential diagnoses are inflammatory myofibroblastic tumors, pseudotumoral tumors, interdigitant dendritic cell sarcomas, lymphomas and undifferentiated carcinomas. Diagnosis is mainly based on pathological examination with immunohistochemical complement.

Recurrence of follicular dendritic cell sarcoma in tonsils after 7 years: a rare case report

Follicular dendritic cell sarcoma (FDCS) is a rare mesenchymal neoplasm. It arises from not only from lymph nodes but also from extra nodal tissues, either as acquired lymphoid tissue or as part of the organized constitutive lymphoid tissue. We here report this rare entity which developed again after 7 years post tonsillectomy in a 5-year old male patient. Patient underwent radical tonsillectomy and adjuvant treatment is awaited. Differential diagnosis includes large cell lymphoma, peripheral nerve sheath tumor, extracranial meningioma, malignant melanoma, metastatic carcinoma, ectopic thymoma, malignant fibrous histiocytoma, and interstitial reticulum cell sarcoma. Currently, the management of FDCS includes the therapeutic guidelines similar to that of high-grade soft tissue sarcomas that is complete surgical resection of the lesion with possibility of adjuvant radiotherapy and/or chemotherapy. The ideal combination of management of FDCS has yet to be defined.

A combination of 5/6-nephrectomy and unilateral ureteral obstruction model accelerates progression of remote organ fibrosis in chronic kidney disease.

Chronic kidney disease (CKD) involves progressive renal fibrosis, which gradually reduces kidney function and often causes various complications in extrarenal tissues. Therefore, we investigated fibrogenesis in extrarenal tissues (heart, liver, and lungs) in different experimental CKD models, such as the 5/6-nephrectomy (5/6 Nx), unilateral ureteral obstruction (UUO), and a combination (2/3 Nx + UUO). We evaluated the degree of fibrogenesis in kidneys and extrarenal tissues by histological analysis and quantification of fibrosis-related gene and protein expression. To elucidate the fibrosis mechanisms observed in 2/3 Nx + UUO mice, we evaluated the effect of indoxyl sulfate (IS), a typical uremic toxin accumulated in CKD, and transforming growth factor-β (TGF-β), a fibrosis-related factor, on fibrosis using human hepatoma (HepG2) and RAW264.7 cells. A significant decline in renal function was observed in the 5/6 Nx and 2/3 Nx + UUO models, whereas a significant increase in renal fibrosis was observed only in the obstructed kidneys. Notable amount of fibrosis was induced in the liver and heart in the 2/3 Nx + UUO model, with the induction of macrophage infiltration and increased tissue IS and TGF-β levels. In agreement with the results of in vivo experiments, co-stimulation with IS, TGF-β, and macrophage-conditioned medium increased the expression of fibrogenic genes in HepG2 cells. We demonstrated that the 2/3 Nx + UUO model induced both loss of renal function and renal fibrosis in the earlier stages, providing a novel CKD model that induces remote organ fibrosis in a shorter time.

A Selective Mineralocorticoid Receptor Blocker, Esaxerenone, Attenuates Vascular Dysfunction in Diabetic C57BL/6 Mice.

Pharmacological blockade of mineralocorticoid receptors (MRs) is a potential therapeutic approach to reduce cardiovascular complications since MRs play a crucial role in cardiovascular regulation. Recent studies suggest that MR antagonists affect several extrarenal tissues, including vessel function. We investigated the effect of a novel nonsteroidal selective MR blocker, esaxerenone, on diabetes-induced vascular dysfunction. Diabetes was induced by a single dose of streptozotocin in 8-week-old male C57BL/6 mice. Esaxerenone (3 mg/kg/day) or a vehicle was administered by gavage to diabetic mice for 3 weeks. Metabolic parameters, plasma aldosterone levels, and parameters related to renal function were measured. Endothelium-dependent or -independent vascular responses of the aortic segments were analyzed with acetylcholine or sodium nitroprusside, respectively. Human umbilical vein endothelial cells (HUVECs) were used for the in vitro study. Induction of diabetes elevated plasma aldosterone level (P＜0.05) and impaired endothelium-dependent vascular relaxation (P＜0.05). The administration of esaxerenone ameliorated the endothelial dysfunction (P＜0.01) without the alteration of metabolic parameters, blood pressure, and renal function. Esaxerenone improved the eNOSSer1177 phosphorylation in the aorta obtained from diabetic mice (P＜0.05) compared with that in the vehicle-treated group. Furthermore, a major MR agonist, aldosterone, decreased eNOSSer1177 phosphorylation and increased eNOSThr495 phosphorylation in HUVECs, which recovered with esaxerenone. Esaxerenone ameliorated the endothelium-dependent vascular relaxation caused by aldosterone in the aortic segments obtained from C57BL/6 mice (P＜0.001). Esaxerenone attenuates the development of diabetes-induced endothelial dysfunction in mice. These results suggest that esaxerenone has potential vascular protective effects in individuals with diabetes.

Vitamin D in hypoparathyroidism: insight into pathophysiology and perspectives in clinical practice.

Hypoparathyroidism (HypoPT) is a rare endocrine disorder characterized by the absence or insufficient parathyroid hormone production resulting in chronic hypocalcemia. Complications of HypoPT include perturbation of several target organs. The conventional treatment consists of the administration of active vitamin D, namely calcitriol. Regarding vitamin D status, few data are available, mostly in HypoPT subjects supplemented with parent vitamin D. In addition, perturbation of vitamin D metabolism has been poorly investigated, as well as the contribution of altered vitamin D status on the clinical expression of the disease. The most recent consensus on the management of chronic HypoPT suggests the baseline evaluation of serum 25-hydroxy-vitamin D [25(OH)D] and supplementation with parent vitamin D with the aim to achieve and maintain serum 25(OH)D levels in the range of 30-50 ng/mL. The rationale for using supplementation with parent vitamin D (either ergocalciferol or cholecalciferol) in HypoPT would be to provide sufficient 25(OH)D substrate to the residual 1-α-hydroxylase activity, thus ensuring its conversion to active vitamin D in renal and extra-renal tissues. More data from experimental and clinical studies are needed for better assessing how these mechanisms may significantly influence metabolic control in HypoPT and eventually skeletal and extra-skeletal manifestation of the disease. Finally, future data will clarify how the currently available parent vitamin D compounds (ergocalciferol, cholecalciferol, calcifediol) would perform in addressing these specific issues.

Hormonal imbalance in patients with chronic renal failure in the pre-dialysis and dialysis periods (part1)

The review describes the problem of studying progressive changes of hormones concentrations (parathyroid hormone, insulin, somatotropin, prolactin) in patients with chronic renal failure on the pre-dialysis and dialysis stages. The pathogenetic relationships between kidney function deterioration and hormone concentrations as well as changes of their biological effects were evaluated. Parathyroid hormone is considered as an uraemic toxin, since its concentration in the blood begins to increase when the glomerular filtration rate decreases below 50 ml/min. All stages of chronic kidney disease are accompanied by disorders of calcium-phosphorus metabolism. Prolonged excess of parathyroid hormone leads to bone loss and to the progression to secondary hyperparathyroidism that is a frequent complication in patients with the later stages of chronic renal failure and, especially, in those on dialysis treatment. The elevation of insulin level in chronic renal failure is the consequence of progressive decrease in glomerular filtration rate and insulin excretion by proximal tubules. So, it results in insulin half-life prolongation. Long-term dialysis therapy eliminates factors that reduce the degradation of insulin by extrarenal tissues, which results in an improvement of their insulin sensitivity. Experimental and clinical studies have shown that an excess of somatotropin can adversely affect the kidneys that leads to glomerular hyperfiltration and the progression to glomerulosclerosis. The risk of possible side effects on kidneys should be taken into account when prescribing recombinant human insulin-like growth factor. The prolactin concentration is usually increased in chronic kidney disease due to reduced clearance and increased secretion. Hyperprolactinemia manifests as galactorrhea and hypogonadism. Dialysis therapy can’t normalize the increased concentration of prolactin. Modern options for pathogenetic treatment of endocrine disorders in patients with chronic renal failure are outlined in this article. It was found that kidneys play an important role in regulating hormones concentrations in the blood. Endocrine disorders are one of the most important components of the uraemic syndrome, which requires further clinical studies, aimed on the searching of better treatment strategies and prevention of hormonal imbalance on the pre-dialysis and dialysis stages of chronic kidney disease.

Analysis Homestay Management During Covid-19 Pandemic in Kebangsaan Tourism Village Wonorejo

This study aims to analyze the homestay management during the covid-19 pandemic in the Kebangsaan Tourism Village Wonorejo. The existence of homestays is very important to support tourist activities, such as extending the length of stay of tourists. Homestay was developed from 2017 and continues to increase and has an impact on the local economy. Since the COVID-19 pandemic, management has implemented health protocols in accordance with government recommendations. Therefore, it is necessary to analyze the management of homestays during the covid-19 pandemic. This research was conducted with a descriptive-qualitative approach. The data analysis technique used the POAC management theory (planning, organizing, actuating, and controlling) to obtain the results of homestay management during the Covid-19 pandemic in the application of CHSE. The results of the study show that the management of homestays in the Kebangsaan Tourism Village during the Covid-19 pandemic has not implemented CHSE. Some have taken basic infection prevention measures such as encouraging frequent hand washing, providing hand sanitizer, urging employees to stay home if they are sick, preparing extra tissues and trash containers in public areas, increasing time in room maintenance. In addition, several managements have also implemented hygiene and environmental safety, such as: cleaning surfaces that look dirty followed by disinfection and performing additional cleaning on all frequently touched areas. Management has not implemented social distancing. Management needs training and socialization in implementing CHSE for homestay management.

PS-BPB06-8: THE NON-STEROIDAL MINERALOCORTICOID RECEPTOR ANTAGONIST, FINERENONE, REDUCES VASCULAR PATHOLOGY AND INFLAMMATION IN PRE-CLINICAL DIABETIC RETINOPATHY

Objective: Diabetic retinopathy is the leading cause of vision loss in the working population across the globe. Diabetes has devastating effects on the retinal vasculature, and can lead to vision-threatening vascular leakage, odema, and neovascularization. Current treatments such as anti-vascular endothelial growth factor (VEGF) are administered directly into the eye late in the disease process but are ineffective in about 50% of patients. Hypertension is a risk factor for the development of diabetic retinopathy and the renin-angiotensin aldosterone system contributes to disease pathogenesis. Here we aimed to determine if finerenone, a non-steroidal mineralocorticoid receptor (MR) antagonist, reduced vascular pathology and inflammation in animal models of diabetic retinopathy. Methods: To study diabetic retinopathy, streptozotocin diabetes was induced in female hypertensive transgenic Ren-2 rats overexpressing renin in extra-renal tissues. Diabetic Ren-2 rats were randomised to receive finerenone (10 mg/kg/day) by oral gavage or the angiotensin converting enzyme inhibitor perindopril (10 mg/kg/day) in drinking water for 12 weeks. As retinal neovascularization does not develop in diabetic rodents, the oxygen-induced retinopathy (OIR) model was studied. Litters of C57BL/6J mice were exposed to high oxygen (75%) between postnatal days 7 to 12 and then room air until postnatal day 18. Mouse pups were randomised to be OIR controls, or treated with finerenone (5 mg/kg/day) by intraperitoneal injection between postnatal days 7 to 18. Comparisons were made to mice exposed to room air. Damage to the blood-retinal barrier was evaluated by measuring the gliosis of macroglial Müller cells which confer barrier properties, vascular leakage (albumin ELISA), neovascularization, and the expression of VEGF. Inflammation was evaluated by the quantitation of retinal microglia and the expression of pro-inflammatory mediators. Results: Systolic blood pressure was similar between untreated non-diabetic (174.2 ± 6.3 mmHG) and diabetic (184.2 ± 8.5 mmHG) Ren-2 rats. Finerenone (138.5 ± 5.01 mmHG, P < 0.0001) and perindopril (106 ± 2.9 mmHG, P < 0.0001) reduced systolic blood pressure compared to untreated diabetic rats. In diabetic rats, finerenone reduced the gliosis of retinal Müller cells, vascular leakage, microglial density and the expression of VEGF, ICAM-1 and IL-1β;. In diabetic rats, perindopril did not reduce retinal VEGF levels, microglial density and the expression of pro-inflammatory mediators. In OIR mice, finerenone reduced retinal neovascularization, vascular leakage and microglial density. Conclusion: Our findings placed in the context of the FIDELIO-DKD trial reporting the benefits of finerenone on renal and cardiovascular outcomes in diabetic kidney disease, indicate the potential of finerenone to be an effective orally administered treatment for diabetic retinopathy.

Soluble ACE2 Is Filtered into the Urine.

ACE2 is a key enzyme in the renin-angiotensin system (RAS) capable of balancing the RAS by metabolizing angiotensin II (AngII). First described in cardiac tissue, abundance of ACE2 is highest in the kidney, and it is also expressed in several extrarenal tissues. Previously, we reported an association between enhanced susceptibility to hypertension and elevated renal AngII levels in global ACE2-knockout mice. To examine the effect of ACE2 expressed in the kidney, relative to extrarenal expression, on the development of hypertension, we used a kidney crosstransplantation strategy with ACE2-KO and WT mice. In this model, both native kidneys are removed and renal function is provided entirely by the transplanted kidney, such that four experimental groups with restricted ACE2 expression are generated: WT→WT (WT), KO→WT (KidneyKO), WT→KO (SystemicKO), and KO→KO (TotalKO). Additionally, we used nanoscale mass spectrometry-based proteomics to identify ACE2 fragments in early glomerular filtrate of mice. Although significant differences in BP were not detected, a major finding of our study is that shed or soluble ACE2 (sACE2) was present in urine of KidneyKO mice that lack renal ACE2 expression. Detection of sACE2 in the urine of KidneyKO mice during AngII-mediated hypertension suggests that sACE2 originating from extrarenal tissues can reach the kidney and be excreted in urine. To confirm glomerular filtration of ACE2, we used micropuncture and nanoscale proteomics to detect peptides derived from ACE2 in the Bowman's space. Our findings suggest that both systemic and renal tissues may contribute to sACE2 in urine, identifying the kidney as a major site for ACE2 actions. Moreover, filtration of sACE2 into the lumen of the nephron may contribute to the pathophysiology of kidney diseases characterized by disruption of the glomerular filtration barrier.

LBODP016 Lithocholic Acid Induced 25-hydroxyvitamin D-1alpha Hydroxylase (cyp27b1) Expression And Activities In Vitro And In Vivo

Abstract Our previous study reported that oleanolic acid, a natural agonist for bile acid membrane receptor Takeda G protein coupled receptor 5 (TGR5), could exert bone protective effects, increase circulating 1, 25(OH)2D3 level and induce renal CYP27B1 expression and activities. However, it is unclear if bile acids, as the endogenous agonist of bile acid receptors, could regulate CYP27B1 expression and activities in kidney and bone tissues. The present study aimed to characterize the effects of lithocholic acid (LCA), a secondary bile acid, on CYP27B1 expression and activities in vitro using human proximal tubule (HKC-8) and osteosarcoma (MG-63) and in vivo using mature (6-month-old) male C57BL/6 mice. HKC-8 and MG-63 cells were treated with either LCA (10-10 to 10-5 M), PTH (10-7M), or its vehicle for 4 hours. For MG-63 cells, LCA significantly induced CYP27B1 protein expression at 10-10 and 10-9M (P<0. 001) and CYP27B1 mRNA expression at 10-9M (P<0. 001). For HKC-8 cells, LCA significantly induced CYP27B1 protein expression at 10-10 to 10-6M (P<0. 01) and CYP27B1 mRNA expression at 10-10M (P<0. 001). In addition, LCA significantly induced CYP27B1 activity in MG-63 cells (10-10M) (P<0. 05) and in HKC-8 cells (10-6M) (P<0. 01). For the in vivo study, male C57BL/6 mice were fed with 25mg/kg LCA or its vehicle for three consecutive days and sacrificed 5 hours after the last gavage. Serum, kidney, and iliac crests were collected for subsequent analysis. LCA significantly induced circulating 1,25(OH)2D3 levels (P<0. 01) and CYP27B1 protein expression in both kidney and iliac crests in mice (P <0. 05). In addition, the time-dependent change (2, 5, 8 hours) in serum 1,25(OH)2D3 levels were determined upon feeding the male mice with a single dose of 25mg/kg LCA. LCA significantly increased serum 1,25(OH)2D3 levels in male mice at 2 and 5 hours (P <0. 01 and P <0. 05, respectively), and serum 1,25(OH)2D3 levels returned to basal level by 8 hours. LCA significantly increased CYP27B1 protein expression in kidney in mice at 8 hours after treatment (vs. control group, P<0. 05). In summary, our results showed that LCA significantly induced circulating 1,25(OH)2D3 levels, possibly by its actions to induce CYP27B1 expressions and activities in renal and extra-renal (bone) tissues. The study indicated that bile acids and other TGR5 ligands might play an important role in the regulation of vitamin D metabolism. Presentation: No date and time listed

Physics Informed Neural Networks (PINN) for Low Snr Magnetic Resonance Electrical Properties Tomography (MREPT).

Electrical properties (EPs) of tissues facilitate early detection of cancerous tissues. Magnetic resonance electrical properties tomography (MREPT) is a technique to non-invasively probe the EPs of tissues from MRI measurements. Most MREPT methods rely on numerical differentiation (ND) to solve partial differential Equations (PDEs) to reconstruct the EPs. However, they are not practical for clinical data because ND is noise sensitive and the MRI measurements for MREPT are noisy in nature. Recently, Physics informed neural networks (PINNs) have been introduced to solve PDEs by substituting ND with automatic differentiation (AD). To the best of our knowledge, it has not been applied to MREPT due to the challenges in using PINN on MREPT as (i) a PINN requires part of ground-truth EPs as collocation points to optimize the network's AD, (ii) the noisy input data disrupts the optimization of PINNs despite the noise-filtering nature of NNs and additional denoising processes. In this work, we propose a PINN-MREPT model based on a canonical analytic MREPT model. A reference padding layer with known EPs was added to surround the region of interest for providing additive collocation points. Moreover, an optimizable diffusion coefficient was embedded in the analytic MREPT model used in the PINN-MREPT. The noise robustness of the proposed PINN-MREPT for single-sample reconstruction was tested by using numerical phantoms of human brain with extra tumor-like tissues at different noise levels. The results of numerical experiments show that PINN-MREPT outperforms two typical numerical MREPT methods in terms of reconstruction accuracy, sensitivity to the extra tissues, and the correlations of line profiles in the regions of interest. The advantage of the PINN-MREPT is shown by the results of an experiment on phantom measurement, too. Moreover, it is found that the diffusion term plays an important role to achieve a noise-robust PINN-MREPT. This is an important step moving forward to a clinical application of MREPT.

Feasibility and safety of laparoscopic nephrectomy in emphysematous pyelonephritic kidneys

Background: Emphysematous pyelonephritis (EPN) is a serious suppurative infection of renal and extrarenal tissues. Most of them end up in nephrectomy, either in an emergency setting or delayed, which is increasingly favored. Laparoscopic nephrectomy is difficult in the post-EPN state due to dense adhesion around the kidney. Aims and Objectives: In this study, we want to check the feasibility of laparoscopic nephrectomy in EPN kidneys. To the best of our knowledge, our series is the single largest series as of today with 10 cases of laparoscopic nephrectomies in EPN kidneys. Materials and Methods: All patients of EPN who underwent laparoscopic nephrectomy at Institute of Kidney Diseases and Research Centre, Ahmedabad, between December 2017 and October 2019 were taken as study subjects and retrospectively reviewed. Patients in whom conservative management was successful were excluded from the study. All patients were diverted initially, either ultrasound/computed tomography-guided percutaneous nephrostomy (PCN) insertion or double J stenting, with or without PCN along with antibiotics and supportive therapy. Delayed transperitoneal laparoscopic nephrectomies were performed for poorly functioning EPN kidneys in follow-up renal scans. Results: A total of 10 patients underwent nephrectomy in this period. The male-to-female ratio was 3:7. As per Huang and Tseng’s EPN classification, one patient had Class 2, four patients had Class 3a, and five patients had Class 3b EPN disease. Mean operative time was 192 min and mean blood loss was 206 ml. Two cases had intraoperative complications managed laparoscopically. No conversion to open nephrectomy or mortality was seen. The drain was placed for a mean period of 3.1 days. The mean hospital stay was 4.8 days. Conclusion: Laparoscopy is safe and feasible for nephrectomy in EPN kidneys. Experience with laparoscopy and operating on pyelonephritis cases is important to reduce complications and extend the advantages of laparoscopy in these groups of patients. An increase in class does not increase the degree of difficulty during laparoscopy.

KLHL3 deficiency in mice ameliorates obesity, insulin resistance, and nonalcoholic fatty liver disease by regulating energy expenditure

Obesity is a growing global epidemic that can cause serious adverse health consequences, including insulin resistance (IR) and nonalcoholic fatty liver disease (NAFLD). Obesity development can be attributed to energy imbalance and metabolic inflexibility. Here, we demonstrated that lack of Kelch-like protein 3 (KLHL3) mitigated the development of obesity, IR, and NAFLD by increasing energy expenditure. KLHL3 mutations in humans cause Gordon’s hypertension syndrome; however, the role of KLHL3 in obesity was previously unknown. We examined differences in obesity-related parameters between control and Klhl3−/− mice. A significant decrease in body weight concomitant with fat mass loss and improved IR and NAFLD were observed in Klhl3−/− mice fed a high-fat (HF) diet and aged. KLHL3 deficiency inhibited obesity, IR, and NAFLD by increasing energy expenditure with augmentation of O2 consumption and CO2 production. Delivering dominant-negative (DN) Klhl3 using adeno-associated virus into mice, thereby dominantly expressing DN-KLHL3 in the liver, ameliorated diet-induced obesity, IR, and NAFLD. Finally, adenoviral overexpression of DN-KLHL3, but not wild-type KLHL3, in hepatocytes revealed an energetic phenotype with an increase in the oxygen consumption rate. The present findings demonstrate a novel function of KLHL3 mutation in extrarenal tissues, such as the liver, and may provide a therapeutic target against obesity and obesity-related diseases.