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Pamabrom is a diuretic that is effective in treating premenstrual syndrome and primary dysmenorrhea. The aim of this study was to examine the effect of metformin and modulators of the opioid receptor-nitric oxide (NO)-cyclic guanosine monophosphate (cGMP)-[K.sup.+] channel pathway on the local antinociception induced by pamabrom. The rat paw 1% formalin test was used to assess the effects. Rats were treated with local administration of pamabrom (200-800 [micro]g/paw) or indomethacin (200800 [micro]g/paw). The antinociception of pamabrom or indomethacin was evaluated with and without the local pretreatment of the blockers. Local administration of pamabrom and indomethacin produced dose-dependent antinociception during the second phase of the test. Local pretreatment of the paws with naloxone (50 [micro]g/paw), L-nitro-arginine methyl ester (10-100 [micro]g/paw), or 1H-(1,2,4)-oxadiazolo[4,2-a]quinoxalin-1-one (10-100 [micro]g/paw) reverted the antinociception induced by local pamabrom, but not of indomethacin. Similarly, the [K.sup.+] channel blockers glibenclamide, glipizide, 4-aminopyridine, tetraethylammonium, charybdotoxin, or apamin reverted the pamabrom-induced antinociception, but not of indomethacin. Metformin significantly blocked the antinociception of pamabrom and indomethacin. Our data suggest that pamabrom could activate the opioid receptor-NO-cGMP-[K.sup.+] channel pathway to produce its peripheral antinociception in the formalin test. Likewise, a biguanide-dependent mechanism could be activated by pamabrom and indomethacin to generate antinociception. Key words: pamabrom, antinociception, nitric oxide, cGMP, opioid receptor, metformin, [K.sup.+] channels, Introduction Premenstrual syndrome (PMS) is a disorder of cyclic recurrence that encompasses a set of repetitive symptoms, which can occur a few days before menstruation and decrease or disappear with [...]

The objective of the present study was to scrutinize the effect of nitric oxide (NO), cyclic GMP (cGMP), potassium channel blockers, and metformin on the citral-produced peripheral antinociception. The rat paw 1% formalin test was used to assess nociception and antinociception. Rats were treated with local peripheral administration of citral (10- 100 [micro]g/paw). The antinociception of citral (100 [micro]g/paw) was evaluated with and without the local pretreatment of naloxone, NG-L-nitro-arginine methyl ester (L-NAME, a NO synthesis inhibitor), 1H-(1,2,4)-oxadiazolo(4,2-a)quinoxalin-1-one (ODQ a soluble guanylyl cyclase inhibitor), metformin, opioid receptors antagonists, and [K.sup.+] channel blockers. Injection of citral in the rat paw significantly decreased the nociceptive effect of formalin administration during the two phases of the test. Local pretreatment of the paws with L-NAME and ODQ did not reduced the citral-induced antinociception. Glipizide or glibenclamide ([K.sub.ir]6.1-2; ATP-sensitive [K.sup.+] channel blockers), tetraethylammonium or 4-aminopyridine ([K.sub.V]; voltage-gated [K.sup.+] channel blockers), charybdotoxin ([K.sub.ca]1.1; big conductance calcium-activated [K.sup.+] channel blocker), apamin ([K.sub.ca]2.1- 3; small conductance [Ca.sup.2+]-activated [K.sup.+] channel antagonist), or metformin, but not the opioid antagonists, reduced the antinociception of citral. Citral produced peripheral antinociception during both phases of the formalin test. These effects were due to the activation of [K.sup.+] channels and a biguanide-dependent mechanism. Key words: antinociception, citral, nociception, [K.sup.+] channels, metformin, formalin test, rat, peripheral effect. La presente etude avait pour objectif de scruter l'effet de l'oxyde nitrique (NO), du GMPc, de bloqueurs des canaux potassiques et de la metformine sur l'antinociception peripherique entrainee par le citral. Nous avons evalue la nociception et l'antinociception a l'aide du test a la formaline (1 %) sur la patte de rat. Nous avons procede a l'administration peripherique locale de citral (10-100 [micro]g/patte) chez les rats. Nous avons evalue le pouvoir antinociceptif du citral (100 [micro]g/patte) avec et sans administration locale prealable de naloxone, de L-NAME (pour << NG-L-nitro-arginine methyl ester >>, un inhibiteur de la synthese du NO), d'ODQ (pour << 1H-(1,2,4)-oxadiazolo(4,2-a)quinoxalin-1-one >>, un inhibiteur de la guanylyl cyclase soluble), de metformine, d'antagonistes des recepteurs des opiaces et de bloqueurs des canaux [K.sup.+]. L'injection de citral dans la patte des rats entrainait une diminution notable de l'effet nociceptif de l'administration de formaline au cours des deux phases du test. L'administration locale prealable de L-NAME et d'ODQ n'a pas entraine de diminution de l'antinociception entrainee par le citral. Le glipizide ou le glibenclamide ([K.sub.ir]6.1-2; des bloqueurs des canaux [K.sup.+] sensibles a l'ATP), le tetraethylammonium ou la 4-aminopyridine ([K.sub.V]; des bloqueurs des canaux [K.sup.+] dependants du voltage), la charybdotoxine ([K.sub.ca]1.1; un bloqueur des canaux [K.sup.+] a conductance elevee actives par le calcium) ou l'apamine ([K.sub.Ca]2.1-3; un antagoniste de canaux [K.sup.+] a faible conductance actives par les ions [Ca.sup.2+]), ou la metformine ont entraine une diminution de l'effet antinociceptif du citral, mais pas les antagonistes des recepteurs des opiaces. Le citral a entraine une antinociception peripherique au cours des deux phases du test a la formaline. Ces effets etaient lies a l'activation des canaux [K.sup.+] et a un mode d'action dependant des biguanides. [Traduit par la Redaction] Mots-cles : antinociception, citral, nociception, canaux [K.sup.+], metformine, test a la formaline, rat, effet peripherique., Introduction Citral, also known as geranial or neral, is an aromatic terpene derivative that can be found in different plants and citrus fruits. Citral is found as a clear yellowish [...]

The aim of this study was to examine if the peripheral antinociception of [alpha]-bisabolol involves the participation of nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) synthesis followed by [K.sup.+] channel opening in the formalin test. Wistar rats were injected in the dorsal surface of the right hind paw with formalin (1%). Rats received a subcutaneous injection into the dorsal surface of the paw of vehicles or increasing doses of [alpha]-bisabolol (100-300 [micro]g/paw). To determine whether the peripheral antinociception induced by [alpha]-bisabolol was mediated by either the opioid receptors or the NO-cGMP-[K.sup.+] channels pathway, the effect of pretreatment (10 min before formalin injection) with the appropriate vehicles, naloxone, naltrexone, [N.sup.G]-nitro-L-arginine methyl ester (L-NAME), 1H-[1,2,4]-oxadiazolo[4,2-a]quinoxalin-1-one (ODQ), glibenclamide, glipizide, apamin, charybdotoxin, tetraethylammonium, or 4-aminopyridine on the antinociceptive effects induced by local peripheral [alpha]-bisabolol (300 [micro]g/paw) were assessed. [alpha]-Bisabolol produced antinociception during both phases of the formalin test. [alpha]-Bisabolol antinociception was blocked by L-NAME, ODQ, and all the [K.sup.+] channels blockers. The peripheral antinociceptive effect produced by [alpha]-bisabolol was not blocked by the opioid receptor inhibitors. [alpha]-Bisabolol was able to active the NO-cGMP-[K.sup.+] channels pathway to produce its antinoceptive effect. The participation of opioid receptors in the peripheral local antinociception induced by [alpha]-bisabolol is excluded. Key words: [alpha]-bisabolol, nociception, nitric oxide, cGMP, [K.sup.+] channels. Cette etude visait a evaluer si les effets de la synthese d'oxyde nitrique (NO) et de guanosine monophosphate cyclique (GMPc) suivie de l'ouverture des canaux [K.sup.+] participent aux effets antinociceptifs peripheriques de l'[alpha]-bisabolol dans le test a la formaline. Nous avons injecte de la formaline (1 %) dans la face dorsale de la patte posterieure droite de rats Wistar. Nous avons injecte par voie sous- cutanee les vehicules ou de l'[alpha]-bisabolol a doses croissantes dans la face dorsale de la patte des rats (100-300 [micro]g/patte). En vue d'etablir si l'antinociception peripherique engendree par l'[alpha]-bisabolol etait mediee par les recepteurs aux opiaces ou par la voie de signalisation NO-cGMP--canaux [K.sup.+], nous avons evalue l'effet de l'administration prealable (10 min avant l'injection de formaline) des vehicules appropries, de naloxone, de naltrexone, de L-NAME, d'ODQ de glibenclamide, de glipizide, d'apamine, de charybdotoxine, de tetraethylammonium ou de 4-aminopyridine sur les effets antinociceptifs engendres par l'administration locale d'[alpha]-bisabolol en peripherie (300 [micro]g/patte). L'[alpha]-bisabolol entrainait une antinociception aux cours des deux phases du test a la formaline. Nous avons inhibe l'antinociception par l'[alpha]-bisabolol a l'aide de L-NAME, d'ODQ et de tous les bloqueurs des canaux [K.sup.+]. L'effet antinociceptif peripherique produit par l'[alpha]-bisabolol n'etait pas inhibe par les inhibiteurs des recepteurs aux opiaces. L'[alpha]-bisabolol permettait d'activer la voie de signalisation NO-cGMP--canaux [K.sup.+] pour produire son effet antinociceptif. Nous excluons la participation des recepteurs aux opiaces dans l'antinociception peripherique locale engendree par l'[alpha]- bisabolol. [Traduit par la Redaction] Mots-cles: [alpha]-bisabolol, nociception, oxyde nitrique, GMPc, canaux [K.sup.+]., Introduction [alpha]-Bisabolol (IUPAC name: (2 R)-6-methyl-2-[(1 R)-4-methylcyclohex3-en-1- yl] hept-5-en-2-ol) is a plant-derived, natural, unsaturated, monocyclic sesquiterpene alcohol. [alpha]-Bisabolol is the primary constituent from several aromatic plants such as German chamomile [...]

The aim of this study was to examine if the peripheral antinociceptive effects of the opioid agonist/antagonist nalbuphine and buprenorphine involve the sequential participation of nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) synthesis followed by [K.sup.+] channel opening in the formalin test. Wistar rats (180-220 g) were injected in the dorsal surface of the right hind paw with formalin (1%). Rats received a subcutaneous (s.c.) injection into the dorsal surface of the paw of vehicles or increasing doses of nalbuphine (50-200 [micro]g/paw) or buprenorphine (1-5 [micro]g/paw) 20 min before formalin injection into the paw. Nalbuphine antinociception was reversed by the s.c. injection into the paw of the inhibitor of NO synthesis ([N.sup.G]-nitro-L-arginine methyl ester (L-NAME)), by the inhibitor of guanylyl cyclase (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ)), by the [K.sub.ir]6.1-2, ATP-sensitive [K.sup.+] channel inhibitors (glibenclamide and glipizide), by the [K.sub.Ca]2.1-3, small conductance [Ca.sup.2+]-activated [K.sup.+] channel blocker (apamin), by the [K.sub.Ca]1.1, large conductance [Ca.sup.2+]-activated [K.sup.+] channel blocker (charybdotoxin), and by the KV, voltage-dependent [K.sup.+] channel inhibitors (4-aminopyridine (4-AP) and tetraethylammonium chloride (TEA)). The antinociceptive effect produced by buprenorphine was blocked by the s.c. injection of 4-AP and TEA but not by L-NAME, ODQ, glibenclamide, glipizide, apamin, or charybdotoxin. The present results provide evidence for differences in peripheral mechanisms of action between these opioid drugs. Key words: opioids, potassium channels, nitric oxide, nociception, rat formalin test. Cette etude portait sur les effets antinociceptifs peripheriques d'opiaces agoniste et antagoniste, la nalbuphine et la buprenorphine, jouant un role dans la participation sequentielle de la synthese d'oxyde nitrique (NO) et de la guanosine monophosphate cyclique (GMPc), suivie de l'ouverture des canaux [K.sup.+] a l'aide du test a la formaline. Nous avons injecte de la formaline (1 %) dans la face dorsale de la patte posterieure de rats Wistar (180-220 g). Les rats ont recu dans la face dorsale de la patte une injection par voie sous-cutanee (s.-c.) de vehicule ou de nalbuphine (50-200 [micro]g/patte) ou de buprenorphine (1-5 [micro]g/patte) a doses croissantes 20 min avant l'injection de formaline dans la patte. L'antinociception par la nalbuphine s'inversait avec l'injection par voie s.-c. de l'inhibiteur de la synthese de NO (methyl ester de [N.sup.G]-nitro-L-arginine (L-NAME)), d'un inhibiteur de la guanylyl cyclase (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ)), d'inhibiteurs des canaux [K.sup.+] sensibles a l'ATP, [K.sub.ir]6.1-2 (glibenclamide et glipizide), d'un inhibiteur des canaux [K.sup.+] actives par le [Ca.sup.2+] de faible conductance [K.sub.Ca]2.1-3 (apamine), d'un inhibiteur des canaux [K.sup.+] actives par le [Ca.sup.2+]de forte conductance, [K.sub.Ca]1.1 (charybdotoxine) et d'inhibiteurs des canaux [K.sup.+] dependants du voltage, KV (4-aminopyridine (4-AP) et chlorure de tetraethylammonium (TEA)). L'effet antinociceptif produit par la buprenorphine etait inhibe par l'injection par voie s.-c. de 4-AP et de TEA, mais pas avec le L-NAME, l'ODQ, le glibenclamide, le glipizide, l'apamine ou la charybdotoxine. Les presents resultats apportent des donnees probantes quant aux differences entre ces opiaces selon leurs modes d'action peripheriques. [Traduit par la Redaction] Mots-cles : opiaces, canaux potassiques, oxyde nitrique, nociception, test a la formaline chez le rat., Introduction The opioid system modulates several physiological processes, including analgesia, stress responses, and neuroendocrine function. Opioid drugs are some of the most efficacious analgesics used in humans (Ballantyne and Sullivan [...]

Simple Summary: In diabetic nephropathy, the injury of renal cells such as podocytes worsens the glomerular filtration process due to the cells' loss and detachment from the basal membrane. Intra- and intercellular communications in cells are partially mediated by the Rab3A/Rab27A system. We have observed that silencing the Rab3A/Rab27A system in podocytes experiencing glucose overload has a differential impact on differentiation, cytoskeleton organisation, apoptosis, CD63 distribution, and miRNA levels. Generally, when cells are subjected to glucose stress and the Rab system is silenced, we observed a reduction in the detrimental cell processes. These results suggest that the Rab3A/Rab27A system is a key participant in podocyte injury and vesicular traffic regulation in diabetic nephropathy. Diabetic nephropathy is a major complication in diabetic patients. Podocytes undergo loss and detachment from the basal membrane. Intra- and intercellular communication through exosomes are key processes for maintaining function, and the Rab3A/Rab27A system is an important counterpart. Previously, we observed significant changes in the Rab3A/Rab27A system in podocytes under glucose overload, demonstrating its important role in podocyte injury. We investigated the implication of silencing the Rab3A/Rab27A system in high glucose-treated podocytes and analysed the effect on differentiation, apoptosis, cytoskeletal organisation, vesicle distribution, and microRNA expression in cells and exosomes. For this, we subjected podocytes to high glucose and transfection through siRNAs, and we isolated extracellular vesicles and performed western blotting, transmission electron microscopy, RT-qPCR, immunofluorescence and flow cytometry assays. We found that silencing RAB3A and RAB27A generally leads to a decrease in podocyte differentiation and cytoskeleton organization and an increase in apoptosis. Moreover, CD63-positive vesicles experienced a pattern distribution change. Under high glucose, Rab3A/Rab27A silencing ameliorates some of these detrimental processes, suggesting a differential influence depending on the presence or absence of cellular stress. We also observed substantial expression changes in miRNAs that were relevant in diabetic nephropathy upon silencing and glucose treatment. Our findings highlight the Rab3A/Rab27A system as a key participant in podocyte injury and vesicular traffic regulation in diabetic nephropathy. [ABSTRACT FROM AUTHOR]

Despite considerable progress in our understanding of systemic lupus erythematosus (SLE) pathophysiology, patient diagnosis is often deficient and late, and this has an impact on disease progression. The aim of this study was to analyze non-coding RNA (ncRNA) packaged into exosomes by next-generation sequencing to assess the molecular profile associated with renal damage, one of the most serious complications of SLE, to identify new potential targets to improve disease diagnosis and management using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. The plasma exosomes had a specific ncRNA profile associated with lupus nephritis (LN). The three ncRNA types with the highest number of differentially expressed transcripts were microRNAs (miRNAs), long non-coding RNAs (lncRNAs) and piwi-interacting RNAs (piRNAs). We identified an exosomal 29-ncRNA molecular signature, of which 15 were associated only with LN presence; piRNAs were the most representative, followed by lncRNAs and miRNAs. The transcriptional regulatory network showed a significant role for four lncRNAs (LINC01015, LINC01986, AC087257.1 and AC022596.1) and two miRNAs (miR-16-5p and miR-101-3p) in network organization, targeting critical pathways implicated in inflammation, fibrosis, epithelial–mesenchymal transition and actin cytoskeleton. From these, a handful of potential targets, such as transforming growth factor-β (TGF-β) superfamily binding proteins (activin-A, TGFB receptors, etc.), WNT/β-catenin and fibroblast growth factors (FGFs) have been identified for use as therapeutic targets of renal damage in SLE. [ABSTRACT FROM AUTHOR]

Non-coding RNA (ncRNA)-mediated targeting of various genes regulates the molecular mechanisms of the pathogenesis of hypertension (HTN). However, very few circulating long ncRNAs (lncRNAs) have been reported to be altered in essential HTN. The aim of our study was to identify a lncRNA profile in plasma and plasma exosomes associated with urinary albumin excretion in HTN by next-generation sequencing and to assess biological functions enriched in response to albuminuria using GO and KEGG analysis. Plasma exosomes showed higher diversity and fold change of lncRNAs than plasma, and low transcript overlapping was found between the two biofluids. Enrichment analysis identified different biological pathways regulated in plasma or exosome fraction, which were implicated in fatty acid metabolism, extracellular matrix, and mechanisms of sorting ncRNAs into exosomes, while plasma pathways were implicated in genome reorganization, interference with RNA polymerase, and as scaffolds for assembling transcriptional regulators. Our study found a biofluid specific lncRNA profile associated with albuminuria, with higher diversity in exosomal fraction, which identifies several potential targets that may be utilized to study mechanisms of albuminuria and cardiovascular damage. [ABSTRACT FROM AUTHOR]

Autoimmune diseases (ADs) are characterized by the activation of the immune system against self-antigens. More common in women than in men and with an early onset, their incidence is increasing worldwide, and this, combined with their chronic nature, is contributing to an enlarged medical and economic burden. Conventional immunosuppressive agents are designed to alleviate symptoms but do not constitute an effective therapy, highlighting a need to develop new alternatives. In this regard, mesenchymal stem cells (MSCs) have demonstrated powerful immunosuppressive and regenerative effects. MSC-derived extracellular vesicles (MSC-EVs) have shown some advantages, such as less immunogenicity, and are proposed as novel therapies for ADs. In this review, we summarize current perspectives on therapeutic options for ADs based on MSCs and MSC-EVs, focusing particularly on their mechanism of action exerted through their non-coding RNA (ncRNA) cargo. A complete state-of-the-art review was performed, centralized on some of the most severe ADs (rheumatoid arthritis, autoimmune type 1 diabetes mellitus, and systemic lupus erythematosus), giving evidence that a promising field is evolving to overcome the current knowledge and provide new therapeutic possibilities centered on MSC-EVs and their role as ncRNA delivery vehicles for AD gene therapy. [ABSTRACT FROM AUTHOR]

Artificial Intelligence is creating a paradigm shift in health care, with phenotyping patients through clustering techniques being one of the areas of interest. Objective: To develop a predictive model to classify heart failure (HF) patients according to their left ventricular ejection fraction (LVEF), by using available data from Electronic Health Records (EHR). Subjects and methods: 2854 subjects over 25 years old with a diagnosis of HF and LVEF, measured by echocardiography, were selected to develop an algorithm to predict patients with reduced EF using supervised analysis. The performance of the developed algorithm was tested in heart failure patients from Primary Care. To select the most influentual variables, the LASSO algorithm setting was used, and to tackle the issue of one class exceeding the other one by a large amount, we used the Synthetic Minority Oversampling Technique (SMOTE). Finally, Random Forest (RF) and XGBoost models were constructed. Results: The full XGBoost model obtained the maximum accuracy, a high negative predictive value, and the highest positive predictive value. Gender, age, unstable angina, atrial fibrillation and acute myocardial infarct are the variables that most influence EF value. Applied in the EHR dataset, with a total of 25,594 patients with an ICD-code of HF and no regular follow-up in cardiology clinics, 6170 (21.1%) were identified as pertaining to the reduced EF group. Conclusion: The obtained algorithm was able to identify a number of HF patients with reduced ejection fraction, who could benefit from a protocol with a strong possibility of success. Furthermore, the methodology can be used for studies using data extracted from the Electronic Health Records. [ABSTRACT FROM AUTHOR]

Kidney injury in hypertension and diabetes entails, among in other structures, damage in a key cell of the glomerular filtration barrier, the podocyte. Podocytes are polarized and highly differentiated cells in which vesicular transport, partly driven by Rab GTPases, is a relevant process. The aim of the present study was to analyze Rab GTPases of the Rab-Rabphilin system in human immortalized podocytes and the impact of high glucose and angiotensin II. Furthermore, alterations of the system in urine cell pellets from patients with hypertension and diabetes were studied. Apoptosis was analyzed in podocytes, and mRNA level quantification, Western blot analysis, and immunofluorescence were developed to quantify podocyte-specific molecules and Rab-Rabphilin components (Rab3A, Rab27A, and Rabphilin3A). Quantitative RTPCR was performed on urinary cell pellet from patients. The results showed that differentiated cells had reduced protein levels of the Rab-rabphillin system compared with undifferentiated cells. After glucose overload and angiotensin II treatment, apoptosis was increased and podocyte-specific proteins were reduced. Rab3A and Rab27A protein levels were increased under glucose overload, and Rabphilin3A decreased. Furthermore, this system exhibited higher levels under stress conditions in a manner of angiotensin II dose and time treatment. Immunofluorescence imaging indicated different expression patterns of podocyte markers and Rab27A under treatments. Finally, Rab3A and Rab27A were increased in patient urine pellets and showed a direct relationship with albuminuria. Collectively, these results suggest that the Rab-Rabphilin system could be involved in the alterations observed in injured podocytes and that a mechanism may be activated to reduce damage through the vesicular transport enhancement directed by this system. [ABSTRACT FROM AUTHOR]

Circular RNAs (circRNAs) are a class of non-coding RNAs that regulate gene expression by acting as competitive endogenous RNAs (ceRNAs) and modulating gene transcription. Several studies support the implication of circRNAs in a variety of human diseases, but research on the role of circRNAs in systemic lupus erythematosus (SLE) is lacking. In a study recently published in Clinical Science (2018), Zhang et al. identified hsa circ 0012919 as a potential biomarker of disease activity in SLE patients. The authors observed different circRNA expression between SLE patients and healthy controls, an association with clinical variables and with the abnormal DNA methylation present in SLE CD4+ T cells. Finally, Zhang et al. demonstrated that hsa circ 0012919 acts as a miRNA sponge for miR-125a-3p, regulating the gene expression of targets RANTES and KLF13 that are involved in the physiology and pathophysiology of acute and chronic inflammatory processes. These findings support the role of circRNAs in the pathophysiology of SLE. [ABSTRACT FROM AUTHOR]

Identifying new markers of disease flares in lupus nephritis (LN) that facilitate patient stratification and prognosis is important. Therefore, the aim of the present study was to analyze whether urinary SIRT1 expression was altered in LN and whether SIRT1 values in urine could be valuable biomarker of disease activity. In a cohort study, urinary pellets from 40 patients diagnosed with systemic lupus erythematosus (SLE) were analyzed. Clinical measures of lupus activity were assessed. The expression of SIRT1 was quantified by quantitative PCR (qRT-PCR) and immunoblot, then compared between patients with active lupus nephritis, in remission and healthy controls. Association with lupus activity and renal histological features was also analyzed. A significant increase in SIRT1 mRNA levels in patients with active LN was observed compared with those in remission (P=0.02) or healthy controls (P=0.009). In addition, SIRT-1 protein levels were also augmented in LN group than remission (P=0.029) and controls (P=0.001). A strong association was found between SIRT1 expression with anti-dsDNA in SLE and in patients with LN. In addition, histological features in LN biopsies were related with SIRT1, increasing its expression in proliferative forms. Finally, SIRT1 expression values showed a strong discriminatory power of renal injury in SLE. Our study demonstrated an altered urinary expression of SIRT1 and a strong association with disease activity in LN patients, being a valuable marker of renal injury. These results showed the role of the SIRT1 pathway in the SLE pathogenesis. [ABSTRACT FROM AUTHOR]

Objectives To investigate the association between IL18RAP and body mass index (BMI) and obesity and to verify the effect of a polymorphism in the microRNA136 (MIR136) IL18RAP binding region. Design We analysed samples from two Spanish cross-sectional studies, VALCAR (Spanish Mediterranean coast) and Hortega (Spanish centre). These studies aimed at analysing cardiovascular risk and development of cardiovascular disease in the general population. Both populations correspond to regions with different characteristics. Setting Five IL18RAP single nucleotide polymorphisms were selected using the SYSNPs web tool and analysed by oligonucleotide ligation assay (SNPlex). For the MIR136 functional study, cells were transfected with plasmids containing different rs7559479 polymorphism alleles and analysed by luciferase reporter assays. Participants 1970 individuals (Caucasian, both genders): VALCAR (468) and Hortega (1502). Results rs2293225, rs2272127 and rs7559479 showed the following associations: rs7559479 G allele correlated with a higher obesity risk (P=0.01; OR=1.82; 95% CI 1.15 to 2.87 for the VALCAR group; P=0.033; OR=1.35; 95% CI 1.03 to 1.79 for the Hortega population) and higher body mass index (BMI) values (P=0.0045; P=0.1 for VALCAR and Hortega, respectively); a significant association with obesity (P=0.0024, OR=1.44, 95% CI 1.14 to 1.82) and increased BMI values (P=0.008) was found when considering both populations together. rs2293225 T allele was associated with lower obesity risk (P=0.036; OR=0.60; 95% CI 0.35 to 0.96) and lower BMI values (P=0.0038; OR=1.41) while the rs2272127 G allele was associated with lower obesity risk (P=0.028; OR=0.66; 95% CI 0.44 to 0.97) only in the VALCAR population. A reporter assay showed that the presence of the A allele in rs7559479 was associated with increased MIR136 binding to IL18RAP. Conclusions Our results suggest that polymorphisms in IL18RAP influence susceptibility to obesity. We demonstrated that the A allele in rs7559479 increases MIR136 binding, which regulates IL-18 system activity. [ABSTRACT FROM AUTHOR]

Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disease that affects multiple organs. Currently, therapeutic molecules present adverse side effects and are only effective in some SLE patient subgroups. Extracellular vesicles (EV), including exosomes, microvesicles and apoptotic bodies, are released by most cell types, carry nucleic acids, proteins and lipids and play a crucial role in cell-to-cell communication. EVs can stimulate or suppress the immune responses depending on the context. In SLE, EVs can work as autoadjuvants, enhance immune complex formation and maintaining inflammation state. Over the last years, EVs derived from mesenchymal stem cells and antigen presenting cells have emerged as cell-free therapeutic agents to treat autoimmune and inflammatory diseases. In this review, we summarize the current therapeutic applications of extracellular vesicles to regulate immune responses and to ameliorate disease activity in SLE and other autoimmune disorders. [ABSTRACT FROM AUTHOR]

Context:Heliopsis longipes(A. Gray) Blake (Asteraceae), a plant native to Mexico, is used in traditional medicine as analgesic and microbicide. The main component in theH. longipesethanolic extract (HLEE) is affinin, as determined by HPLC/UV–visible and NMR measurement. To date, there is no documented evidence on the spermicidal activity of this extract. Objective: The objective of this study was to assessin vitrothe effectiveness of HLEE as spermicide. Materials and methods: The spermicidal activity of HLEE was evaluated by the Sander–Cramer assay. Spermatozoa were incubated for 20 s with HLEE in concentrations ranging from 75 to 2000 µg/mL to determine the minimum effective concentration (MEC) value. The 50% effective concentration (EC50) of HLEE was estimated by assaying serial dilutions from the MEC. Additionally, sperms were incubated with 125, 250, or 500 µg/mL of HLEE to evaluate the viability and the integrity of sperm membrane. Lipid peroxidation was assessed by the thiobarbituric acid reactive substances assay. Results: HLEE caused an inhibition of 100% in spermatozoa motility at a MEC value of 2000 µg/mL; the EC50value was 125 µg/mL. Additionally, exposure to HLEE at 125, 250, or 500 µg/mL for 30 min decreased sperm viability to 27%, 8%, and 2% of the control value, respectively, and significantly increased the percentage of sperms with structurally disorganized membrane. HLEE also increased significantly the level of lipid peroxidation in sperms with respect to controls. Discussion and conclusion: The results demonstrate the spermicidal activity of HLEEin vitroand suggest that this action is caused by oxidative damage and alterations in the spermatozoal membrane. [ABSTRACT FROM PUBLISHER]

Background and Aims Familial hypercholesterolemia (FH) is associated with higher levels of inflammatory mediators such as chemokines, which contribute to an increased risk of premature atherosclerosis in these patients. We studied the response of chemokines related to early atherosclerotic processes during an oral unsaturated fat load test (OFLT) in patients with heterozygous FH and compared this response to normolipidemic and normoglycemic subjects. Methods Blood samples were taken from 12 FH patients and 20 healthy controls with a similar age, gender distribution, and body mass index. Plasma chemokine levels were determined in both groups in a fasting state and at 2, 4, 6, and 8 h after an OFLT using human cytokine multiplex kits (Linco) and a Luminex LABScan™100 system. Results In the fasting state MIP-1β, MIP-1α, IP-10, IFN-γ, MCP-1, and IL-8 were significantly increased in the FH group compared to the healthy controls ( p <0.05). In addition, a significant decrease in postprandial chemokine plasma values were found in the FH group compared to fasting values after the OFLT. In normolipidemic nondiabetic controls no significant changes were found in the postprandial state. Conclusions There was a postprandial decrease in chemokines related to early atherosclerotic processes after an OFLT in FH patients. These results confirm the influence of dietary patterns in this group of patients. [ABSTRACT FROM AUTHOR]

There is increased interest in using microRNAs (miRNAs) as biomarkers in different diseases. Present in body fluids, it is controversial whether or not they are mainly enclosed in exosomes, thus we studied if urinary miRNAs are concentrated inside exosomes and if the presence of systemic lupus erythematosus with or without lupus nephritis modifies their distribution pattern. We quantified specific miRNAs in urine of patients with systemic lupus erythematosus (n = 38) and healthy controls (n = 12) by quantitative reverse-transcription PCR in cell-free urine, exosome-depleted supernatant and exosome pellet obtained by ultracentrifugation. In control group, miR-335* and miR-302d were consistently higher in exosomes than in exosome-depleted supernatant, and miR-200c and miR-146a were higher in cell-free fraction. In lupus patients, all urinary miRNAs tested were mainly in exosomes with lower levels outside them (p<0.05 and p<0.01, respectively). This pattern is especially relevant in patients with active lupus nephritis compared to the control group or to the SLE patients in absence of lupus nephritis, with miR-146a being the most augmented (100-fold change, p<0.001). Among the exosomal miRNAs tested, only the miR-146a discriminates the presence of active lupus nephritis. In conclusion, urinary miRNAs are contained primarily in exosomes in systemic lupus erythematosus, and the main increment was found in the presence of active lupus nephritis. These findings underscore the attractiveness of exosomal miRNAs in urine, a non-invasive method, as potential renal disease markers. [ABSTRACT FROM AUTHOR]

Non-coding RNA (ncRNA), released into circulation or packaged into exosomes, plays important roles in many biological processes in the kidney. The purpose of the present study is to identify a common ncRNA signature associated with early renal damage and its related molecular pathways. Three individual libraries (plasma and urinary exosomes, and total plasma) were prepared from each hypertensive patient (with or without albuminuria) for ncRNA sequencing analysis. Next, an RNA-based transcriptional regulatory network was constructed. The three RNA biotypes with the greatest number of differentially expressed transcripts were long-ncRNA (lncRNA), microRNA (miRNA) and piwi-interacting RNA (piRNAs). We identified a common 24 ncRNA molecular signature related to hypertension-associated urinary albumin excretion, of which lncRNAs were the most representative. In addition, the transcriptional regulatory network showed five lncRNAs (LINC02614, BAALC-AS1, FAM230B, LOC100505824 and LINC01484) and the miR-301a-3p to play a significant role in network organization and targeting critical pathways regulating filtration barrier integrity and tubule reabsorption. Our study found an ncRNA profile associated with albuminuria, independent of biofluid origin (urine or plasma, circulating or in exosomes) that identifies a handful of potential targets, which may be utilized to study mechanisms of albuminuria and cardiovascular damage. [ABSTRACT FROM AUTHOR]

Small Rab GTPases, the largest group of small monomeric GTPases, regulate vesicle trafficking in cells, which are integral to many cellular processes. Their role in neurological diseases, such as cancer and inflammation have been extensively studied, but their implication in kidney disease has not been researched in depth. Rab3a and its effector Rabphillin-3A (Rph3A) expression have been demonstrated to be present in the podocytes of normal kidneys of mice rats and humans, around vesicles contained in the foot processes, and they are overexpressed in diseases with proteinuria. In addition, the Rab3A knockout mice model induced profound cytoskeletal changes in podocytes of high glucose fed animals. Likewise, RphA interference in the Drosophila model produced structural and functional damage in nephrocytes with reduction in filtration capacities and nephrocyte number. Changes in the structure of cardiac fiber in the same RphA-interference model, open the question if Rab3A dysfunction would produce simultaneous damage in the heart and kidney cells, an attractive field that will require attention in the future. [ABSTRACT FROM AUTHOR]

Exosomes, nanometer-sized lipid-bilayer-enclosed extracellular vesicles (EVs), have attracted increasing attention due to their inherent ability to shuttle proteins, lipids and genes between cells and their natural affinity to target cells. Their intrinsic features such as stability, biocompatibility, low immunogenicity and ability to overcome biological barriers, have prompted interest in using exosomes as drug delivery vehicles, especially for gene therapy. Evidence indicates that exosomes play roles in both immune stimulation and tolerance, regulating immune signaling and inflammation. To date, exosome-based nanocarriers delivering small molecule drugs have been developed to treat many prevalent autoimmune diseases. This review highlights the key features of exosomes as drug delivery vehicles, such as therapeutic cargo, use of targeting peptide, loading method and administration route with a broad focus. In addition, we outline the current state of evidence in the field of exosome-based drug delivery systems in systemic lupus erythematosus (SLE), evaluating exosomes derived from various cell types and engineered exosomes. [ABSTRACT FROM AUTHOR]

Sirtuins have become important players in renal damage in hypertension and diabetes, but their value as biomarkers is poorly assessed. The aims of the study were to evaluate the levels of sirtuin1 (SIRT1), and two miRNAs that regulate SIRT1 expression in hypertensive patients with incipient renal damage with and without diabetes. We quantified urinary SIRT1 and claudin 1 (CLDN1) mRNA and miR34-a and miR-200a levels by quantitative real-time polymerase chain reaction (RT-qPCR) from patients and in cultured podocytes treated with high glucose and angiotensin II. Western blot and fluorescence analyses were also performed. We found decreased SIRT1 levels in patients with increased urinary albumin excretion (UAE), the lowest with diabetes presence, and a strong association with UAE, discriminating incipient renal damage. In vitro experiments also showed SIRT1 overall decreases in podocyte cultures under treatment conditions. In urine samples, miR-34a was reduced and miR-200a increased, both related to UAE levels. However, both miRNAs were generally increased in podocyte cultures under high glucose and angiotensin-II treatment. These results show a significant urinary SIRT1 decrease in albuminuric hypertensive patients, strongly associated with albuminuria, suggesting that SIRT1 could be a potential and non-invasive method to assess incipient renal damage in hypertensive patients. [ABSTRACT FROM AUTHOR]

Exosomes, nanometer-sized lipid-bilayer-enclosed extracellular vesicles (EVs), have attracted increasing attention due to their inherent ability to shuttle proteins, lipids and genes between cells and their natural affinity to target cells. Their intrinsic features such as stability, biocompatibility, low immunogenicity and ability to overcome biological barriers, have prompted interest in using exosomes as drug delivery vehicles, especially for gene therapy. Evidence indicates that exosomes play roles in both immune stimulation and tolerance, regulating immune signaling and inflammation. To date, exosome-based nanocarriers delivering small molecule drugs have been developed to treat many prevalent autoimmune diseases. This review highlights the key features of exosomes as drug delivery vehicles, such as therapeutic cargo, use of targeting peptide, loading method and administration route with a broad focus. In addition, we outline the current state of evidence in the field of exosome-based drug delivery systems in systemic lupus erythematosus (SLE), evaluating exosomes derived from various cell types and engineered exosomes.

Despite the wide application of carvacrol (CAR) in different biological and medical areas, there is still insufficient electrophysiological data on the mechanisms of action of CAR, particularly in the pregnant uterine function. The aim of this study was to evaluate the in vitro tocolytic effect of CAR on the contractility of isolated pregnant rat uterus in the presence of a calcium channel antagonist (nifedipine) and a cyclooxygenase inhibitor (indomethacin). The uteri were isolated from pregnant Wistar rats at 16-18 days of pregnancy and suspended in an isolated organ bath chamber containing a Ringer's physiological solution and aerated with 95% O 2 and 5% CO 2 . Samples were used in functional tests to evaluate the inhibitory effect of CAR at increasing concentrations on the rhythmic spontaneous, oxytocin-induced phasic, K + -induced tonic, and Ca 2+ -induced contractions. The differences in inhibitory concentration-50 and E max among the compounds were determined using the one-way ANOVA followed by a post hoc Student-Newman-Keuls or Bonferroni test, in all casesP < 0.05 was considered statistically significant. Nifedipine was used as positive controls where required. CAR caused a significant concentration-dependent inhibition of the uterine contractions induced by the pharmaco- and electro-mechanic stimuli. We showed that the inhibitory effects of CAR depends on the type of muscle contraction stimuli, and that it acts stronger in spontaneous rhythmic activity and in contractions of isolated rat uterus induced by Ca 2+ . Nifedipine was more potent than CAR and indomethacin on the uterine contractility (P < 0.05), but none of them was more effective than nifedipine. Therefore, the tocolytic effect induced by CAR was associated with the blockade of the calcium channels in the pregnant rat uterus. This property placed CAR as a potentially safe and effective adjuvant agent in cases of preterm labor, an area of pharmacological treatment that requires urgent improvement., Competing Interests: None