Fluorescentna in situ hibridizacija za dokazivanje virusa klasične svinjske kuge i praćenje njegove lokalizacije u zaraženim tkivima.

Classical swine fever (CSF) may be diagnosed by various conventional techniques but nucleic acid based methods have gained popularity because of their high sensitivity and specificity. For detection of viral RNA in infected tissues, a fluorescent in-situ hybridization (FISH) based method was developed using a 308 bp biotinylated DNA probe targeting E2/NS2 gene of CSFV. In-situ RNA/DNA hybrids were detected with fluorescently labeled Streptavidin conjugate and viewed under a fluorescent microscope. Viral nucleic acids could be demonstrated in the lymphoid tissues such as the spleen and lymph nodes collected from CSFV infected animals. Infected macrophages and reticuloendothelial cells revealed bright intracytoplasmic fluorescence. In conclusion, the fluorescent in situ hybridization technique used in the study provided an effective means of detection and localization of CSFV nucleic acids.Klasična svinjska kuga može se dijagnosticirati različitim uobičajenim metodama, ali se, zbog visoke osjetljivosti i specifičnosti, najčešće rabe metode zasnovane na dokazivanju specifične nukleinske kiseline. Za dokaz virusne RNA u zaraženom tkivu u ovom je radu razvijena fluorescentna in situ hibridizacija (FISH) uporabom biotinom obilježene DNA probe od 308 bp za ciljni gen E2/NS2 virusa klasične svinjske kuge. RNA/DNA hibridne molekule bile su dokazane uporabom konjugata obilježenog streptavidinom pretragom fluorescentnim mikroskopom. Virusna nukleinska kiselina bila je dokazana u limfoidnim tkivima odnosno u slezeni i limfnim čvorovima životinja zaraženih virusom. Fluorescencija je jasno bila izražena u citoplazmi zaraženih makrofaga i retikuloendotelijalnih stanica. Može se zaključiti da fluorescentna in situ hibridizacija opisana u ovom radu pruža učinkovit alat za dokaz nukleinske kiseline virusa klasične svinjske kuge i njegove lokalizacije u zaraženim tkivima

Major adverse cardiac and cerebrovascular event and patients' quality of life after endoscopic vein harvesting as compared with open vein harvest (MAQEH): a pilot study.

This is a prospective, comparative, pilot and follow-up (2-year postoperatively) study in patients undergoing coronary artery bypass graft surgery where the long saphenous vein was harvested either by the endoscopic vein harvest (EVH) technique or open vein harvest (OVH) technique. Quality of life (QOL) and major adverse cardiac and cerebrovascular events (MACCE) were assessed

Balanced forced-diuresis compared to control as a reno-protective approach in cardiac surgery: secondary outcome of a randomized controlled trial, assessment of neutrophil gelatinase-associated lipocalin levels

© 2021 The Authors. Published by Springer Nature. This is an open access article available under a Creative Commons licence. The published version can be accessed at the following link on the publisher’s website: https://doi.org/10.1186/s13019-021-01620-wBackground: Neutrophil gelatinase-associated lipocalin (NGAL) is a recognised biomarker for acute kidney injury (AKI).This study investigated the impact of balanced forced-diuresis using RenalGuard® system (RG), in reducing acute kidney injury (AKI) rates and the associated NGAL levels (6-h post-CPB plasma level) post adult cardiac surgery with cardiopulmonary bypass (CPB). Methods: Patients included in the study were at high-risk for AKI post cardiac surgery, namely history of diabetes and/or anaemia, e-GFR 20–60 ml/min/1.73 m2, Logistic EuroScore > 5, anticipated CPB time > 120 min. Patients were randomized to either RG (n = 110) or managed as per current practice (control = 110). RIFLE-defined AKI rate (based on serum creatinine level increase) within first 3 days of surgery and 6-h post CPB NGAL levels were the primary and secondary end-points. Results: Pre and intra-operative characteristics between the two groups were similar (p > 0.05) including the pre-op NGAL levels, the oxygen delivery (ecDO2i) and the carbon dioxide production (ecVCO2i) during CPB. Patients in the RG group had a significantly lower post-operative RIFLE-defined AKI rate compared to control (10% (11/110) v/s 20.9% (23/110), p = 0.03). Overall, median 6-h post CPB NGAL levels in patients with AKI were significantly higher than those who did not develop AKI (211 vs 150 ng/ml, p < 0.001). Patients managed by balanced forced-diuresis had lower post-operative NGAL levels (146 vs 178 ng/ml, p = 0.09). Using previously reported NGAL cut-off level for AKI (142 ng/ml), binary logistic regression analysis confirmed a beneficial effect of the RG system, with an increased risk of AKI of 2.2 times in the control group (OR 2.2, 95% CI 1.14–4.27, p = 0.02). Conclusions: Overall, the 6-h post-CPB plasma NGAL levels were significantly higher in patients who developed AKI. Patients managed with the novel approach of balanced forced-diuresis, provided by the RenalGuard® system, had a lower AKI rate and lower NGAL levels indicating a lesser degree of renal tissue injury. Trial registration ClinicalTrials.gov website, NCT02974946, https://clinicaltrials.gov/ct2/show/NCT02974946.This work was supported by a research grant from RenalGuard Solutions and by the National Institute of Healthcare Research (NIHR), Clinical Research Network, UK [Ref: NIHR CRN No. 32769].Accepted versio

Preserved quality of life in octogenarians at early, mid, and late follow-up intervals irrespective of cardiac procedure.

Cardiac surgery has become established in octogenarians over the past decade. This study assessed the quality of life (QOL) and survival in patients undergoing various cardiac procedures at various time intervals postoperative. Patients older than 80 years at the time of their cardiac procedure were initially included (n = 427). Patients were grouped according to the time interval from their operations namely as within 3 years postoperative (Group A), 3-5 years postoperative (Group B), and older than 5 years postoperative (Group C). Patients who were at least 2 years postoperative and who were still alive were sent the Medical Outcomes Study Short Form 12 Health Survey version 2 QOL questionnaire (n = 308). In results, there were no significant differences in the preoperative characteristics among the groups including type of surgery and logistic Euroscore. There were also no significant differences in the immediate postoperative phase in the complication rates except for renal replacement therapy (P < 0.01). At follow-up, a further 20 patients had died, and for those still alive 87% (61 of 70), 86% (86 of 100), and 74% (87 of 118) of patients returned questionnaire for each group, respectively. There were no significant differences in mental scores (P = 0.3) and physical scores (P = 0.07) among the groups at the various time intervals. This was irrespective of the type of surgery performed on multivariate analysis. Moreover, most octogenarians who underwent cardiac surgery had equivalent or better QOL than expected when compared with the general population of the same age and sex. In conclusion, the QOL in octogenarians undergoing cardiac surgery is preserved, irrespective of the interval from and the type of procedure. Octogenarians enjoy a good QOL, both physical and mental performance, irrespective of the time interval after surgery and the type of cardiac surgery that they have undergone. Based on these data, any type of cardiac surgery should still be an option in this age group including complex cardiac procedures

Reduction in acute kidney injury post cardiac surgery using balanced forced diuresis: a randomized, controlled trial

© 2020 The Authors. Published by Oxford University Press on behalf of the European Association for Cardio-Thoracic Surgery. This is an open access article available under a Creative Commons licence. The published version can be accessed at the following link on the publisher’s website: https://doi.org/10.1093/ejcts/ezaa395OBJECTIVES Our goal was to investigate the efficacy of balanced forced diuresis in reducing the rate of acute kidney injury (AKI) in cardiac surgical patients requiring cardiopulmonary bypass (CPB), using the RenalGuard® (RG) system. METHODS Patients at risk of developing AKI (history of diabetes and/or anaemia; estimated glomerular filtration rate 20–60 ml/min/1.73 m2; anticipated CPB time >120 min; log EuroSCORE > 5) were randomized to the RG system group (n = 110) or managed according to current practice (control = 110). The primary end point was the development of AKI within the first 3 postoperative days as defined by the RIFLE (Risk, Injury, Failure, Loss of kidney function, End-stage renal disease) criteria. RESULTS There were no significant differences in preoperative and intraoperative characteristics between the 2 groups. Postoperative AKI rates were significantly lower in the RG system group compared to the control group [10% (11/110) vs 20.9% (23/110); P = 0.025]. This effect persisted even after controlling for a number of potential confounders (odds ratio 2.82, 95% confidence interval 1.20–6.60; P = 0.017) when assessed by binary logistic regression analysis. The mean volumes of urine produced during surgery and within the first 24 h postoperatively were significantly higher in the RG system group (P < 0.001). There were no significant differences in the incidence of blood transfusions, atrial fibrillation and infections and in the median duration of intensive care unit stays between the groups. The number needed to treat with the RG system to prevent AKI was 9 patients (95% confidence interval 6.0–19.2). CONCLUSIONS In patients at risk for AKI who had cardiac surgery with CPB, the RS RG system significantly reduced the incidence of AKI and can be used safely and reproducibly. Larger studies are required to confirm cost benefits.This work was supported by RenalGuard Solutions; and National Institute of Healthcare Research (NIHR), Clinical Research Network, UK [Ref: NIHR CRN No 32769].Published onlin

Activation of Human Monocytes by Live Borrelia burgdorferi Generates TLR2-Dependent and -Independent Responses Which Include Induction of IFN-β

It is widely believed that innate immune responses to Borrelia burgdorferi (Bb) are primarily triggered by the spirochete's outer membrane lipoproteins signaling through cell surface TLR1/2. We recently challenged this notion by demonstrating that phagocytosis of live Bb by peripheral blood mononuclear cells (PBMCs) elicited greater production of proinflammatory cytokines than did equivalent bacterial lysates. Using whole genome microarrays, we show herein that, compared to lysates, live spirochetes elicited a more intense and much broader transcriptional response involving genes associated with diverse cellular processes; among these were IFN-β and a number of interferon-stimulated genes (ISGs), which are not known to result from TLR2 signaling. Using isolated monocytes, we demonstrated that cell activation signals elicited by live Bb result from cell surface interactions and uptake and degradation of organisms within phagosomes. As with PBCMs, live Bb induced markedly greater transcription and secretion of TNF-α, IL-6, IL-10 and IL-1β in monocytes than did lysates. Secreted IL-18, which, like IL-1β, also requires cleavage by activated caspase-1, was generated only in response to live Bb. Pro-inflammatory cytokine production by TLR2-deficient murine macrophages was only moderately diminished in response to live Bb but was drastically impaired against lysates; TLR2 deficiency had no significant effect on uptake and degradation of spirochetes. As with PBMCs, live Bb was a much more potent inducer of IFN-β and ISGs in isolated monocytes than were lysates or a synthetic TLR2 agonist. Collectively, our results indicate that the enhanced innate immune responses of monocytes following phagocytosis of live Bb have both TLR2-dependent and -independent components and that the latter induce transcription of type I IFNs and ISGs

Subulura brumpti infection - An outbreak in Japanese quails (Coturnix coturnix japonica)

Subulura brumpti infection was observed in Japanese quails and caused heavy production loss up to 10%. Gross lesions were confined to caecum of the affected birds. Pathological changes suggestive of acute cecal hemorrhagic enteritis were recorded. Closer observation of the cecal loop revealed wavy movement with thousands of tiny worms. Based on morphometry, the worms were identified as S. brumpti. Condition was responded to the albendazole treatment efficiently and all the birds were recovered and production of the flock has been improved