Gardnerella vaginalis induces NLRP3 inflammasome-mediated pyroptosis in macrophages and THP-1 monocytes

xlomafota13 shared this article with you from Inoreader Gardnerella vaginalis induces NLRP3 inflammasome-mediated pyroptosis in macrophages and THP-1 monocytes Via Experimental and therapeutic medicine Exp Ther Med. 2021 Oct;22(4):1174. doi: 10.3892/etm.2021.10609. Epub 2021 Aug 13. ABSTRACT The vagina is colonized by a variety of microbes that serve vital roles in the maintenance of vaginal health. The purpose of the present study was to explore the underlying mechanism by which Gardnerella vaginalis (GV) can induce bacterial vaginosis (BV). The viability of primary mouse macrophages and THP-1 cells was detected using a Cell Counting Kit-8 assay. Lactate dehydrogenase and caspase-1 activity in the culture medium of macrophages and THP-1 cells were measured using a colorimetric assay and a caspase-1 activity assay kit, respectively. In the macrophages and THP-1 cells, the levels of TNF-α, IL-1β and IL-18 were detected using ELISA whereas reactive oxygen species (ROS) levels were detected using flow cytometry. The pyroptosis of macrophages and THP-1 cells was detected using calcein-AM/PI double staining. Expression of protein s associated with the nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain-containing protein 3 inflammasome (NLRP3), including NLRP3, apoptosis associated speck-like protein (ASC), caspase-1 and pro-caspase-1, were measured by western blotting and reverse transcription-quantitative PCR. GV significantly inhibited cell viability and increased LDH activity in macrophages and THP-1 cells. In addition, GV markedly promoted the production of TNF-α, IL-1β, IL-18 and ROS by macrophages and THP-1 cells. GV significantly promoted caspase-1 activation-mediated pyroptosis in macrophages and THP-1 cells. Treatment with GV significantly increased the protein and mRNA expression of NLRP3, ASC and caspase-1 in macrophages and THP-1 cells. To conclude, data from the present study suggest that G. vaginalis can induce BV by promoting NLRP3 inflammasome-mediated pyroptosis, which provides one of the molecular mechanisms by which G. vaginalis can induce BV. PMID:34504619 | PMC:PMC8393845 | DOI:10.3892/etm.2021.10609 View on the web Inoreader. Take back control of your news feed. Follow us on Twitter and Facebook.

Gadolinium chloride pre-treatment reduces the inflammatory response and preserves intestinal barrier function in a rat model of sepsis

xlomafota13 shared this article with you from Inoreader Gadolinium chloride pre-treatment reduces the inflammatory response and preserves intestinal barrier function in a rat model of sepsis Via Experimental and therapeutic medicine Exp Ther Med. 2021 Oct;22(4):1143. doi: 10.3892/etm.2021.10577. Epub 2021 Aug 9. ABSTRACT The inflammatory response is closely associated with sepsis occurrence and progression. Damage to the function of the intestinal mucosal barrier is considered to be the ῾initiation factor᾿ for the development of multiple organ dysfunction syndrome, which is the most severe progression of sepsis. The aim of the present study was to investigate whether gadolinium chloride (GdCl3) could alleviate the systemic inflammatory response and protect the function of the intestinal mucosal barrier in a rat model of sepsis. The mechanism underlying this protective effect was also explored. Sprague-Dawley rats were divided into four groups: Sham, sham + GdCl3, cecal ligation and puncture (CLP; a model of sepsis) and CLP + GdCl3. In each group, blood was collected from the abdominal aorta, and intestinal tissue was collected after 6, 12 and 24 h of successful modeling. Levels of tumor necrosis factor-α, interleukin (IL)-6 and IL-1β were determined using ELISA. Western blot analysis was used to determine levels of occludin, tight junction protein ZO-1 (ZO-1), myosin light chain kinase 3 (MLCK), NF-κB and caspase-3 in intestinal tissues. Hematoxylin-eosin staining was used to observe the degree of damage to intestinal tissue. The results indicated that in CLP sepsis model rats treated with GdCl3, the release of systemic and intestinal pro-inflammatory factors was reduced and tissue damage was alleviated when compared with untreated CLP rats. Additionally, the expression of occludin and ZO-1 was increased, while that of NF-κB, MLCK, and caspase-3 was reduced in the CLP + GdCl3 rats compared with the CLP rats. GdCl3 may alleviate systemic and intestinal inflammatory responses and reduce the expression of MLCK through inhibition of the activation of NF-kB. The results of the present study also indicated that GdCl3 promoted the expression of occludin and ZO-1. GdCl3 was also demonstrated to reduce cell apoptosis through the inhibition of caspase-3 expression. PMID:34504589 | PMC:PMC8393272 | DOI:10.3892/etm.2021.10577 View on the web Inoreader. Take back control of your news feed. Follow us on Twitter and Facebook.

Piperazine ferulate prevents high-glucose-induced filtration barrier injury of glomerular endothelial cells

xlomafota13 shared this article with you from Inoreader Piperazine ferulate prevents high-glucose-induced filtration barrier injury of glomerular endothelial cells Via Experimental and therapeutic medicine Exp Ther Med. 2021 Oct;22(4):1175. doi: 10.3892/etm.2021.10607. Epub 2021 Aug 13. ABSTRACT Filtration barrier injury induced by high glucose (HG) levels leads to the development of diabetic nephropathy. The endothelial glycocalyx plays a critical role in glomerular barrier function. In the present study, the effects of piperazine ferulate (PF) on HG-induced filtration barrier injury of glomerular endothelial cells (GEnCs) were investigated and the underlying mechanism was assessed. Immunofluorescence was used to observe the distribution of the glycocalyx as well as the expression levels of syndecan-1 and Zonula occludens-1 (ZO-1). Endothelial permeability assays were performed to assess the effects of PF on the integrity of the filtration barrier. Protein and mRNA expression levels were measured by western blotting and reverse transcription-quantitative PCR analyses, respectively. In vitro experiments revealed that adenosine mon ophosphate-activated protein kinase (AMPK) mediated HG-induced glycocalyx degradation and endothelial barrier injury. PF inhibited the HG-induced endothelial barrier injury and restored the expression levels of heparanase-1 (Hpa-1), ZO-1 and occludin-1 by AMPK. In vivo assays demonstrated that PF reduced the expression levels of Hpa-1, increased the expression levels of ZO-1 and attenuated glycocalyx degradation in the glomerulus. These data suggested that PF attenuated HG-induced filtration barrier injury of GEnC by regulating AMPK expression. PMID:34504620 | PMC:PMC8393711 | DOI:10.3892/etm.2021.10607 View on the web Inoreader. Take back control of your news feed. Follow us on Twitter and Facebook.

Deacetylated Sp1 improves β-glycerophosphate-induced calcification in vascular smooth muscle cells

xlomafota13 shared this article with you from Inoreader Deacetylated Sp1 improves β-glycerophosphate-induced calcification in vascular smooth muscle cells Via Experimental and therapeutic medicine Exp Ther Med. 2021 Oct;22(4):1152. doi: 10.3892/etm.2021.10586. Epub 2021 Aug 10. ABSTRACT The aging of the population has led to an annual increase in the incidence of vascular calcification (VC). Specific protein 1 (Sp1) is a transcriptional activator that serves an important role in VC. The deacetylation of transcription factors represses their binding to the promoters of downstream genes, thereby causing their downregulation. The present study aimed to investigate the role of deacetylated Sp1 in the development of VC. In the present study, western blotting and immunoprecipitation (IP) were performed to detect the protein levels of acetylated Sp1. Western blotting and immunofluorescence staining were used to analyze phenotypic switching in vascular smooth muscle cells (VSMCs). Alizarin red S, alkaline phosphatase (ALP) activity and calcium content assays were used to assess calcium deposition in VSMCs. Western blotting, flow cytomet ry, TUNEL staining and caspase3 activity assay were used to evaluate apoptosis of VSMCs. Chromatin immunoprecipitation (ChIP) assay was used to detect Sp1 binding to the BMP2 promoter. The results indicated that, in a β-glycerophosphate (β-GP)-induced VSMC calcification model, the level of acetylated Sp1 was increased. Western blotting and immunofluorescence staining results showed that, compared with the Sp1 overexpression group (Sp1-WT), deacetylated Sp1 (Sp1-K704A) downregulated the expression of osteogenic markers runt-related transcription factor 2 (Runx2) and bone morphogenetic protein 2 (BMP2), and upregulated the expression of contraction marker α-smooth muscle actin (α-SMA) and calponin 1. In addition, deacetylated Sp1 also reduced the ALP activity and calcium content of calcified VSMCs, and the Alizarin red S assay revealed that the calcium crystallization of Sp1-K704A group was markedly decreased. Western blotting, flow cytometry, TUNEL staining and caspase-3 activity assay were detected to indicate that the B-cell lymphoma 2 (Bcl-2)/Bcl-2-associated X protein ratio was increased, and caspase-3 activity and the apoptotic rate of VSMCs were decreased, in the Sp1-K704A group, as compared with the Sp1-WT group. ChIP assay revealed that Sp1 binding to the BMP2 promoter was downregulated in the Sp1-K704A group, compared with that in theSp1-WT group. In conclusion, a deacetylated mutant of Sp1 decreased Sp1 binding to the BMP2 promoter, thus decreasing apoptosis, phenotypic switching and calcium deposition in calcified VSMCs. This finding may indicate potential therapeutic targets for VC. PMID:34504597 | PMC:PMC8394101 | DOI:10.3892/etm.2021.10586 View on the web Inoreader. Take back control of your news feed. Follow us on Twitter and Facebook.

Cyclophosphamide inhibits the progression of Meniere's disease by reducing the generation of circulating immune complex

xlomafota13 shared this article with you from Inoreader Cyclophosphamide inhibits the progression of Meniere's disease by reducing the generation of circulating immune complex Via Experimental and therapeutic medicine Exp Ther Med. 2021 Oct;22(4):1177. doi: 10.3892/etm.2021.10611. Epub 2021 Aug 13. ABSTRACT Endolymphatic hydrops is a characteristic pathological manifestation of Meniere's disease (MD) that has been previously associated with autoimmunity. Interest in the circulating immune complex (CIC) has increased due to its reported role in the occurrence of MD. The present study aimed to investigate the potential value of serum CIC concentration in the diagnosis of MD and the therapeutic potential of cyclophosphamide (CTX) for the treatment of MD. In the present study, guinea pigs were immunized with isologous crude inner ear antigens to establish an autoimmune MD model. Pure tone audiometry, Vestibular-evoked myogenic potential test, electrocochleography test and auditory brainstem response was applied in this study for assessing the severity of MD in guinea pigs. ELISA was applied to measure CIC, tumor necrosis factor α (TNF-α) and heat shoc k protein 70 (HSP70) expression levels in the serum samples of different groups of patients. Western blotting was applied to detect the protein expression of HSP70 in inner ear tissues in guinea pigs. Hematoxylin and eosin staining was applied to visualize the spiral ganglions in spiral ganglions models. CIC expression in the inner ear was detected by immunohistochemistry. In vivo experiments were performed to confirm the therapeutic effects of CTX in MD. Serum concentrations of CIC, TNF-α and HSP70 were found to be significantly higher in patients with MD, which were also associated with increases in hearing classification and the severity of endolymphatic hydrops. Using a guinea pig MD model, ELISA results revealed significantly increased serum CIC, TNF-α and HSP70 concentrations compared with those in the control group. ABR results showed that the thresholds in the CTX group were notably decreased compared with that in the dexamethasone group, whereas CIC concentrations in the serum were reduced following dexamethasone and CTX treatments compared with those after saline treatment. In the inner ear tissues, the CIC concentration in CTX group was lower than that in the dexamethasone group. Similarly, reductions in HSP70 and TNF-α concentrations was also observed in a similar manner. Immunohistochemistry staining found notably lower CIC deposition in the inner ear tissues following CTX treatment than that in dexamethasone group. Taken together, higher CIC expression can be used as a biomarker for MD diagnosis. The efficacy of CTX in MD was found to be higher compared with that in dexamethasone, which may be associated with the effective inhibition of CIC, HSP70 and TNF-α generation. PMID:34504622 | PMC:PMC8393373 | DOI:10.3892/etm.2021.10611 View on the web Inoreader. Take back control of your news feed. Follow us on Twitter and Facebook.

Effect of the combination of astragaloside IV and Panax notoginseng saponins on pyroptosis and necroptosis in rat models of cerebral ischemia-reperfusion

xlomafota13 shared this article with you from Inoreader Effect of the combination of astragaloside IV and <em>Panax notoginseng</em> saponins on pyroptosis and necroptosis in rat models of cerebral ischemia-reperfusion Via Experimental and therapeutic medicine Exp Ther Med. 2021 Oct;22(4):1123. doi: 10.3892/etm.2021.10557. Epub 2021 Aug 4. ABSTRACT Pyroptosis and necroptosis are closely associated with the mechanism underlying cerebral ischemia-reperfusion (I/R) injury. The combination of astragaloside IV (AST IV) and Panax notoginseng saponins (PNS) has remarkable effects on the alleviation of cerebral I/R damage. However, whether inhibition of pyroptosis and necroptosis is the mechanism underlying the beneficial effects of this drug combination on cerebral I/R injury remains unclear. To explore the effects and mechanisms of drug treatment, middle cerebral artery occlusion was performed to induce I/R injury in rats, which was verified based on neurological deficit score (NDS), infarct volume and H&E staining. Activation of pyroptosis and necroptosis was detected by western blot analysis of associated proteins. The results of the present study demonstrated that treatment with AST IV and PNS, either alone or in combination, significantly reduced the NDS, cerebral infarct volume and cell injury rate in the cerebral cortex of rats. The treatments also improved pathological injury to the cerebral cortex and reduced the levels of proteins associated with pyroptosis and necroptosis. These effects were stronger in the combination drug group compared with groups treated with a single drug alone. The findings of the present study suggested that the combination of AST IV and PNS exhibited stronger neuroprotective effects in I/R injury than either drug alone, and that the underlying mechanism was associated with inhibition of pyroptosis and necroptosis. PMID:34504577 | PMC:PMC8383753 | DOI:10.3892/etm.2021.10557 View on the web Inoreader. Take back control of your news feed. Follow us on Twitter and Facebook.

Auswirkungen der COVID-19-Pandemie auf die logopädische Therapie von Kindern mit einer Sprachentwicklungsstörung

xlomafota13 shared this article with you from Inoreader Auswirkungen der COVID-19-Pandemie auf die logopädische Therapie von Kindern mit einer Sprachentwicklungsstörung Via Laryngo-Rhino-Otologie Laryngorhinootologie DOI: 10.1055/a-1613-5747 Hintergrund Im Dezember 2019 begann sich die COVID-19-Pandemie weltweit auszubreiten und sorgte für massive Einschränkungen im täglichen Leben. Viele Bildungseinrichtungen und logopädische Praxen wurden vorübergehend geschlossen (sog. Lockdown). Kinder mit einer Sprachentwicklungsstörung waren gezwungen, ihre Therapie zu pausieren. Ziel dieser Studie war es zu beschreiben, ob und wie die logopädische Therapie während des Lockdowns durchgeführt wurde und welche psychische Belastung für die betroffenen Eltern damit verbunden war. Material und Methoden Eltern von Kindern mit einer Sprachentwicklungsstörung wurden über die Therapie ihres Kindes während des Lockdowns und über ihre damit verbundenen Ängste und Sorgen befragt. Ergebnisse Bei 17 Patienten wurde die Sprachtherapie während des Lockdowns pausiert, während 20 Patienten ihre Therapie fortsetzen konnten. Kinder, die eine andere Sprache als Deutsch sprachen, hatten ein höheres Risiko, dass ihre Therapie pausiert wurde (Odds Ratio (OR) 5,11; 95 %-Konfidenzintervall (KI) 1,09–32,54). Eltern, deren Kinder während des Lockdowns keine Sprachtherapie erhielten, machten sich mehr Sorgen um die Entwicklung ihres Kindes. Schlussfolgerungen Es gibt kein generelles Konzept, wie Logopädie während eines Lockdowns sicher durchgeführt werden kann. Mögliche Barrieren für die Gesundheitsversorgung könnten während der Pandemie stärker ausgeprägt sein. Die betroffenen Eltern erleben eine höhere psychosoziale Belastung. [...] Georg Thieme Verlag KG Rüdigerstraße 14, 70469 Stuttgart, Germany Article in Thieme eJournals: Table of contents  |  Abstract  |  Full text View on the web Inoreader. Take back control of your news feed. Follow us on Twitter and Facebook.

The development of posttraumatic maxillary sinusitis due to transconjunctival penetration of a foreign body with a fracture of the lower wall of the orbit

xlomafota13 shared this article with you from Inoreader The development of posttraumatic maxillary sinusitis due to transconjunctival penetration of a foreign body with a fracture of the lower wall of the orbit Via Vestnik otorinolaringologii Vestn Otorinolaringol. 2021;86(4):111-115. doi: 10.17116/otorino202186041111. ABSTRACT The article presents a clinical example of the course of posttraumatic acute purulent sinusitis with reactive soft tissue phenomena due to the previous injury of the orbit by a foreign body, the introduction of the latter into orbit and the maxillary sinus result in a fracture of the lower wall of the orbit. A feature of the injury is the penetration of a foreign body through the conjunctiva of the lower eyelid and lower conjunctival fornix, without damaging the skin. This case is professionally interesting for both young doctors and experienced specialists in otolaryngology, ophthalmology, maxillofacial surgery and neurosurgery. Experts, analyzing this clinical example, will be able to correctly diagnose, effectively eliminate the inflammatory process in the maxillary sinus. PMID:34499458 | DOI:10.17116/otorino202186041111 View on the web Inoreader. Take back control of your news feed. Follow us on Twitter and Facebook.

Clinical features of BPPV and their influence on the choice of the doctor's tactics

xlomafota13 shared this article with you from Inoreader Clinical features of BPPV and their influence on the choice of the doctor's tactics Via Vestnik otorinolaringologii Vestn Otorinolaringol. 2021;86(4):4-8. doi: 10.17116/otorino2021860414. ABSTRACT OBJECTIVES: BPPV is the most common cause of recurrent vertigo. Except vertigo attacks main clinical symptoms of BPPV can include autonomic symptoms and imbalance, which sometimes complicate the diagnosis of BPPV. Purpose To evaluate clinical symptoms and management of patients with BPPV before the setting of correct diagnose. MATERIAL AND METHODS: A total of 640 patients (504 (78.8%) women) aged from 20 to 86 years old, mean age 56.43±0.54 years with BPPV were included and diagnosed by roll and Dix-Hallpike tests. Among them 144 (22.5%) patients were inpatient and 496 (77.5%) patients were outpatient. The detailed patient intake comprised the disease onset, the type of dizziness, vertigo triggers, autonomic symptoms, similar attacks in the past and previously made definite diagnosis of BPPV. The period from the appearance of the first symptom s to the correct diagnosis was assessed. RESULTS: The majority of patients (75.3%) consult a neurologist at the initial visit. Only 30.6% of patients had a correct diagnosis within a week of the onset of the disease. Initial BPPV symptoms included persistent dizziness that increased with head turns (38.8%), nausea and vomiting (21.6%), significant increase in blood pressure (13.4%), persistent imbalance while walking (73.4%). Inpatients more frequently had constant continuous dizziness, high blood pressure, severe nausea and vomiting, and the onset of symptoms in the morning when getting out of bed (p<0.05). CONCLUSION: Initial BPPV symptoms may be similar to other diseases. Focusing on medical history and complaints leads to frequent diagnostic errors, unnecessary hospitalization and prolonged treatment of patients. Positional tests are necessary for the correct diagnosis of BPPV. PMID:34499439 | DOI:10.17116/otorino2021860414 View on the web Inoreader. Take back control of your news feed. Follow us on Twitter and Facebook.

The main steps in treatment of the children with otitis media with effusion

xlomafota13 shared this article with you from Inoreader The main steps in treatment of the children with otitis media with effusion Via Vestnik otorinolaringologii Vestn Otorinolaringol. 2021;86(4):13-16. doi: 10.17116/otorino20218604113. ABSTRACT THE AIM: Of the investigation was to establish the standard and improve the treatment of otitis media with effusion (OME) in children. 361 children at age from 11 months to 18 years were inspected after tympanostomy during 2013-2018 years. The main diagnostic methods were: otoscopy, tympanometry, endoscopy, CT. MATERIAL AND METHODS: Treatment takes into consideration the reveal of OME: surgical initially. The tympanostomy preferable place is anterior-inferior quadrant. RESULTS: In cases with cleft palate or reccurence OME long-term tubes and balloonisation of ET are preferable. CONCLUSION: Authors received normalization of the hearing thresholds in 97.6% cases, but after surgery the patients have to be followed-up during 12-24 months. PMID:34499441 | DOI:10.17116/otorino20218604113 View on the web Inoreader. Take back control of your news feed. Follow us on Twitter and Facebook.