Gut immune dysfunction through impaired innate pattern recognition receptor expression and gut microbiota dysbiosis in chronic SIV infection.

HIV targets the gut mucosa early in infection, causing immune and epithelial barrier dysfunction and disease progression. However, gut mucosal sensing and innate immune signaling through mucosal pattern recognition receptors (PRRs) during HIV infection and disease progression are not well defined. Using the simian immunodeficiency virus (SIV)-infected rhesus macaque model of AIDS, we found a robust increase in PRRs and inflammatory cytokine gene expression during the acute SIV infection in both peripheral blood and gut mucosa, coinciding with viral replication. PRR expression remained elevated in peripheral blood following the transition to chronic SIV infection. In contrast, massive dampening of PRR expression was detected in the gut mucosa, despite the presence of detectable viral loads. Exceptionally, expression of Toll-like receptor 4 (TLR4) and TLR8 was downmodulated and diverged from expression patterns for most other TLRs in the gut. Decreased mucosal PRR expression was associated with increased abundance of several pathogenic bacterial taxa, including Pasteurellaceae members, Aggregatibacter and Actinobacillus, and Mycoplasmataceae family. Early antiretroviral therapy led to viral suppression but only partial maintenance of gut PRRs and cytokine gene expression. In summary, SIV infection dampens mucosal innate immunity through PRR dysregulation and may promote immune activation, gut microbiota changes, and ineffective viral clearance

Rapid fabrication of pressure-driven open-channel microfluidic devices in omniphobic RF paper

This paper describes the fabrication of pressure-driven, open-channel microfluidic systems with lateral dimensions of 45-300 microns carved in omniphobic paper using a craft-cutting tool. Vapor phase silanization with a fluorinated alkyltrichlorosilane renders paper omniphobic, but preserves its high gas permeability and mechanical properties. When sealed with tape, the carved channels form conduits capable of guiding liquid transport in the low-Reynolds number regime (i.e. laminar flow). These devices are compatible with complex fluids such as droplets of water in oil. The combination of omniphobic paper and a craft cutter enables the development of new types of valves and switches, such as “fold” valves and “porous switches,” which provide new methods to control fluid flow.Chemistry and Chemical Biolog

Inflammation and Immune-Related Candidate Gene Associations with Acute Lung Injury Susceptibility and Severity: A Validation Study

Introduction: Common variants in genes related to inflammation, innate immunity, epithelial cell function, and angiogenesis have been reported to be associated with risks for Acute Lung Injury (ALI) and related outcomes. We tested whether previously-reported associations can be validated in an independent cohort at risk for ALI. Methods: We identified 37 genetic variants in 27 genes previously associated with ALI and related outcomes. We prepared allelic discrimination assays for 12 SNPs from 11 genes with MAF>0.05 and genotyped these SNPs in Caucasian subjects from a cohort of critically ill patients meeting criteria for the systemic inflammatory response syndrome (SIRS) followed for development of ALI, duration of mechanical ventilation, and in-hospital death. We tested for associations using additive and recessive genetic models. Results: Among Caucasian subjects with SIRS (n = 750), we identified a nominal association between rs2069832 in IL6 and ALI susceptibility (OR$_{adj}$ 1.61; 95% confidence interval [CI], 1.04–2.48, P = 0.03). In a sensitivity analysis limiting ALI cases to those who qualified for the Acute Respiratory Distress Syndrome (ARDS), rs61330082 in NAMPT was nominally associated with risk for ARDS. In terms of ALI outcomes, SNPs in MBL2 (rs1800450) and IL8 (rs4073) were nominally associated with fewer ventilator-free days (VFDs), and SNPs in NFE2L2 (rs6721961) and NAMPT (rs61330082) were nominally associated with 28-day mortality. The directions of effect for these nominal associations were in the same direction as previously reported but none of the associations survived correction for multiple hypothesis testing. Conclusion: Although our primary analyses failed to statistically validate prior associations, our results provide some support for associations between SNPs in IL6 and NAMPT and risk for development of lung injury and for SNPs in IL8, MBL2, NFE2L2 and NAMPT with severity in ALI outcomes. These associations provide further evidence that genetic factors in genes related to immunity and inflammation contribute to ALI pathogenesis

Retinal glycoprotein enrichment by concanavalin a enabled identification of novel membrane autoantigen synaptotagmin-1 in equine recurrent uveitis.

Complete knowledge of autoantigen spectra is crucial for understanding pathomechanisms of autoimmune diseases like equine recurrent uveitis (ERU), a spontaneous model for human autoimmune uveitis. While several ERU autoantigens were identified previously, no membrane protein was found so far. As there is a great overlap between glycoproteins and membrane proteins, the aim of this study was to test whether pre-enrichment of retinal glycoproteins by ConA affinity is an effective tool to detect autoantigen candidates among membrane proteins. In 1D Western blots, the glycoprotein preparation allowed detection of IgG reactions to low abundant proteins in sera of ERU patients. Synaptotagmin-1, a Ca2+-sensing protein in synaptic vesicles, was identified as autoantigen candidate from the pre-enriched glycoprotein fraction by mass spectrometry and was validated as a highly prevalent autoantigen by enzyme-linked immunosorbent assay. Analysis of Syt1 expression in retinas of ERU cases showed a downregulation in the majority of ERU affected retinas to 24%. Results pointed to a dysregulation of retinal neurotransmitter release in ERU. Identification of synaptotagmin-1, the first cell membrane associated autoantigen in this spontaneous autoimmune disease, demonstrated that examination of tissue fractions can lead to the discovery of previously undetected novel autoantigens. Further experiments will address its role in ERU pathology

Patrick’s Point State Park Habitat Restoration Plan

Ongoing efforts at Patrick’s Point State Park (PPSP) seek to restore the ecosystems contained therein. Over time, non-native invasive plant species have become established within the Park. This habitat restoration plan (or Plan) aims to aid in restoring native and sensitive habitat through the removal of non-native invasive species. The Plan will take many different approaches to targeting the invasive non-native plant species depending on the target species and specific site conditions

Standardizacija kontrolnih laboratorijskih metoda u mljekarstvu - Uzimanje uzoraka mlijeka iz kanti i transportnih cisterni

Pravilan postupak uzimanja uzoraka mlijeka je složen i odgovoran rad ako se želi postići realne rezultate analiza. Različita fizikalno-kemijska svojstva pojedinih sastojaka mlijeka i različiti postupci čuvanja mlijeka prouzrokuju različit raspored sastojaka u mlijeku koje miruje duže vrijeme. Miješanje i proporcionalna količina, osnova su za prosječnost i time reprezentativnost uzorka. U radu su sustavno praćeni rezultati analiza mlijeka kod različitih postupaka ručnog miješanja u najčešće upotrebljavanim transportnim posudama, kako bi se pridonijelo raščišćavanju pitanja uspješnosti miješanja mlijeka prije uzimanja uzoraka

Open infrainguinal revascularization for chronic arterial occlusions: techniques and short-time clinical results

Catedra de chirurgie generală semiologie nr. 3, Universitatea de Stat de Medicină și Farmacie „Nicolae Testemiţanu”, Spitalul Clinic Municipal nr.1, Chişinău, Republica Moldova, Al XIII-lea Congres al Asociației Chirurgilor „Nicolae Anestiadi” și al III-lea Congres al Societății de Endoscopie, Chirurgie miniminvazivă și Ultrasonografie ”V.M.Guțu” din Republica MoldovaIntroducere: În pofida implementării active a tehnicilor endovasculare intervențiile arteriale deschise adresate leziunilor ocluzive cronice infrainghinale își păstrează utilitatea, reprezentând uneori chiar unica opțiune de revascularizare a extremității ischemizate. Scopul studiului a fost evidențierea ratei de aplicare în practica cotidiană a diverselor tehnici chirurgicale deschise de revascularizare în caz de ocluzii arteriale cronice infrainghinale și evaluarea rezultatelor clinice precoce ale acestora. Material și metode: Datele medicale referitoare la un grup constituit din 190 de pacienți revascularizați pentru leziuni aterosclerotice infrainghinale în cadrul Clinicii Chirurgie Generală – Semiologie nr.3, IMSP SCM nr.1 (Chișinău), au fost extrase din registrul electronic și supuse analizei retrospective. Rezultate: Vârsta medie a bolnavilor – 67,3±9,8 ani; bărbați – 83,2%. Repartizarea observațiilor conform gradului de ischemie (clasificarea Fontaine): IIB – 59 (31%), III – 52 (27,3%) și IV – 79 (41,5%). Ultrasonografia duplex/TC-angiografia au pus în evidență localizarea preponderentă a leziunilor ocluzive la nivelul arterei femurale (50%). Sediul ocluziei în 10,5% cazuri a fost artera femurală comună, iar în alte 10% – axul distal (trunchiul tibio-peronier, aa.tibiale). Revascularizarea s-a realizat prin: bypass femuropopliteu (46,4%), -infrapopliteu (13,3%) sau -distal (6,3%); endarterectomie cu angioplastie (18,4%); angioplastie cu petic (10,8%); arterializarea sistemului venos (2,5%) sau protezare arterială (1,9%). Rata amputațiilor secundare realizate în termen precoce (30 zile) de la revascularizare – 3,7%. Concluzii: La momentul adresării pacientului leziunile arteriale cronice ocluzive infrainghinale frecvent au caracter extins. Intervențiile deschise cu utilizarea întregului arsenal de tehnici de revascularizare oferă rezultate precoce acceptabile, în special la pacienții fără comorbidități critice.Introduction: Despite the active implementation of endovascular techniques, open arterial reconstructions for infrainguinal chronic occlusive arterial lesions (ICOAL) retain their usefulness, sometimes representing the sole option for revascularization and salvage of the affected extremity. Aim of study was to evaluate the rate of use in routine clinical practice of various open surgical techniques for revascularization in case of ICOAL and to assess their short-term clinical results. Material and methods: Medical data related to a group of 190 patients revascularized for ICOAL at the Department of General Surgery-Semiotics nr.3, Municipal Clinical Hospital no.1 (Chisinau), were extracted from the electronic registry and subjected to retrospective analysis. Results: The mean age of patients – 67.3±9.8 years; males – 83.2%. Distribution of cases according to the grade of ischemia (Fontaine classification): IIB – 59 (31%), III – 52 (27.3%) and IV – 79 (41.5%). Duplex ultrasound/CT-angiography revealed the predominant localization of ICOAL at the level of femoral artery (50%). The occlusion site in 10.5% cases was the common femoral artery, while in another 10% – the leg arteries (tibioperoneal trunk, tibial arteries). Revascularization was achieved by: femoro-popliteal (46.4%), -infrapopliteal (13.3%) or -distal (6.3%) bypasses; endarterectomy with angioplasty (18.4%); patch angioplasty (10.8%); venous arterialization (2.5%) or graft interposition (1.9%). The rate of secondary amputation within 30 days after revascularization was 3.7%. Conclusion: At the moment of patient presentation ICOAL frequently have extensive character. Open surgical procedures with utilization of all spectrum of techniques of revascularization provide acceptable short-term results, especially in patients without critical comorbidities

Electric Stimulation of Neurogenesis Improves Behavioral Recovery After Focal Ischemia in Aged Rats

© Copyright © 2020 Balseanu, Grigore, Pinosanu, Slevin, Hermann, Glavan and Popa-Wagner. The major aim of stroke therapies is to stimulate brain repair and to improve behavioral recuperation after cerebral ischemia. Despite remarkable advances in cell therapy for stroke, stem cell-based tissue replacement has not been achieved yet stimulating the search for alternative strategies for brain self-repair using the neurogenic zones of the brain, the dentate gyrus and the subventricular zone (SVZ). However, during aging, the potential of the hippocampus and the SVZ to generate new neuronal precursors, declines. We hypothesized that electrically stimulation of endogenous neurogenesis in aged rats could increase the odds of brain self-repair and improve behavioral recuperation after focal ischemia. Following stroke in aged animals, the rats were subjected to two sessions of electrical non-convulsive stimulation using ear-clip electrodes, at 7- and 24 days after MCAO. Animal were sacrificed after 48 days. We report that electrical stimulation (ES) stimulation of post-stroke aged rats led to an improved functional recovery of spatial long-term memory (T-maze) but not on the rotating pole or the inclined plane, both tests requiring complex sensorimotor skills. Surprisingly, ES had a detrimental effect on the asymmetric sensorimotor deficit. Histologically, there was a robust increase in the number of doublecortin-positive cells in the dentate gyrus and SVZ of the infarcted hemisphere and the presence of a considerable number of neurons expressing tubulin beta III in the infarcted area. Among the gene that were unique to ES, we noted increases in the expression of seizure related 6 homolog like which is one of the physiological substrate of the β-secretase BACE1 involved in the pathophysiology of the Alzheimer’s disease and Igfbp3 and BDNF receptor mRNAs which has been shown to have a neuroprotective effect after cerebral ischemia. However, ES was associated with a long-term down regulation of cortical gene expression after stroke in aged rats suggesting that gene expression in the peri-infarcted cortical area may not be related to electrical stimulation induced-neurogenesis in the subventricular zone and hippocampus

Differential Patterns of Synaptotagmin7 mRNA Expression in Rats with Kainate- and Pilocarpine-Induced Seizures

Previous studies in rat models of neurodegenerative disorders have shown disregulation of striatal synaptotagmin7 mRNA. Here we explored the expression of synaptotagmin7 mRNA in the brains of rats with seizures triggered by the glutamatergic agonist kainate (10 mg/kg) or by the muscarinic agonist pilocarpine (30 mg/kg) in LiCl (3 mEq/kg) pre-treated (24 h) rats, in a time-course experiment (30 min - 1 day). After kainate-induced seizures, synaptotagmin7 mRNA levels were transiently and uniformly increased throughout the dorsal and ventral striatum (accumbens) at 8 and 12 h, but not at 24 h, followed at 24 h by somewhat variable upregulation within different parts of the cerebral cortex, amigdala and thalamic nuclei, the hippocampus and the lateral septum. By contrast, after LiCl/pilocarpine-induced seizures, there was a more prolonged increase of striatal Synaptotagmin7 mRNA levels (at 8, 12 and 24 h), but only in the ventromedial striatum, while in some other of the aforementioned brain regions there was a decline to below the basal levels. After systemic post-treatment with muscarinic antagonist scopolamine in a dose of 2 mg/kg the seizures were either extinguished or attenuated. In scopolamine post-treated animals with extinguished seizures the striatal synaptotagmin7 mRNA levels (at 12 h after the onset of seizures) were not different from the levels in control animals without seizures, while in rats with attenuated seizures, the upregulation closely resembled kainate seizures-like pattern of striatal upregulation. In the dose of 1 mg/kg, scopolamine did not significantly affect the progression of pilocarpine-induced seizures or pilocarpine seizures-like pattern of striatal upregulation of synaptotagmin7 mRNA. In control experiments, equivalent doses of scopolamine per se did not affect the expression of synaptotagmin7 mRNA. We conclude that here described differential time course and pattern of synaptotagmin7 mRNA expression imply regional differences of pathophysiological brain activation and plasticity in these two models of seizures