Helminth induced monocytosis conveys protection from respiratory syncytial virus infection in mice.

BACKGROUND: Respiratory syncytial virus (RSV) infection in infants is a major cause of viral bronchiolitis and hospitalisation. We have previously shown in a murine model that ongoing infection with the gut helminth Heligmosomoides polygyrus protects against RSV infection through type I interferon (IFN-I) dependent reduction of viral load. Yet, the cellular basis for this protection has remained elusive. Given that recruitment of mononuclear phagocytes to the lung is critical for early RSV infection control, we assessed their role in this coinfection model. METHODS: Mice were infected by oral gavage with H. polygyrus. Myeloid immune cell populations were assessed by flow cytometry in lung, blood and bone marrow throughout infection and after secondary infection with RSV. Monocyte numbers were depleted by anti-CCR2 antibody or increased by intravenous transfer of enriched monocytes. RESULTS: H. polygyrus infection induces bone marrow monopoiesis, increasing circulatory monocytes and lung mononuclear phagocytes in a IFN-I signalling dependent manner. This expansion causes enhanced lung mononuclear phagocyte counts early in RSV infection that may contribute to the reduction of RSV load. Depletion or supplementation of circulatory monocytes prior to RSV infection confirms that these are both necessary and sufficient for helminth induced antiviral protection. CONCLUSIONS: H. polygyrus infection induces systemic monocytosis contributing to elevated mononuclear phagocyte numbers in the lung. These cells are central to an anti-viral effect that reduces the peak viral load in RSV infection. Treatments to promote or modulate these cells may provide novel paths to control RSV infection in high risk individuals.

Cold dispase digestion of murine lungs improves recovery and culture of airway epithelial cells.

Airway epithelial cells (AECs) play a key role in maintaining lung homeostasis, epithelium regeneration and the initiation of pulmonary immune responses. To isolate and study murine AECs investigators have classically used short and hot (1h 37°C) digestion protocols. Here, we present a workflow for efficient AECs isolation and culture, utilizing long and cold (20h 4°C) dispase II digestion of murine lungs. This protocol yields a greater number of viable AECs compared to an established 1h 37°C dispase II digestion. Using a combination of flow cytometry and immunofluorescent microscopy, we demonstrate that compared to the established method, the cold digestion allows for recovery of a 3-fold higher number of CD45-CD31-EpCAM+ cells from murine lungs. Their viability is increased compared to established protocols, they can be isolated in larger numbers by magnetic-activated cell sorting (MACS), and they result in greater numbers of distal airway stem cell (DASC) KRT5+p63+ colonies in vitro. Our findings demonstrate that temperature and duration of murine lung enzymatic digestion have a considerable impact on AEC yield, viability, and ability to form colonies in vitro. We believe this workflow will be helpful for studying lung AECs and their role in the biology of lung.

Early and late outcomes of percutaneous transluminal angioplasty of cephalad arteries.

BACKGROUND: Efficacy of carotid endarterectomy (CEA) in prevention of stroke in patients with carotid artery stenosis has been confirmed in randomised trials. Carotid artery stenting (CAS) is a routine clinical practice and recent results of CAS are not worse than CEA. Moreover, percutaneous transluminal angioplasty (PTA) techniques allow other cephalad arteries to be dilated. AIM: To assess early and long-term outcome of PTA of cephalad arteries and to determine risk factors of early and late major adverse cardiovascular and cerebral events (MACCE). METHODS: The study group consisted of 223 consecutive patients (151 males, 67.7%, mean age 65.3+/-8.6) in whom 256 PTA procedures of cephalad arteries were performed. Two hundred and forty-two internal carotid, 7 common carotid and 15 vertebral arteries were dilated. Thirty-four patients underwent one-stage carotid and coronary procedures, while in 46 patients one-stage carotid and peripheral procedures were performed. Neuroprotection with a distal protection device was used in 51.5% of cases. The procedures were divided into two groups: with high (n=181) and low (n=75) risk of cardiovascular events. Early and late events were recorded and analysed subsequently. RESULTS: In hospital 30-day MACCE occurred in 12 (4.6%) patients, including 7 (2.7%) strokes, 3 (1.1%) myocardial infarctions and two (0.8%) deaths. Transient ischaemic attacks were observed in 8 patients, pulmonary oedema in 3 cases, as well as a single episode of retinal artery embolisation and acute renal insufficiency. The incidence of 30-day MACCE was not significantly higher in the high-risk group (6.07 vs. 1.33%; NS), but the risk of any adverse event was significantly higher (p=0.03). There was no difference in stroke incidence between procedures with or without neuroprotection (2.27 vs. 3.22%; NS). There was no difference in risk of MACCE between angioplasty of cephalad artery and one-stage cephalad and coronary artery angioplasty procedure (3.6 vs. 5.5%; NS). During 50.3+/-20 months of follow-up there were 16 (7.1%) deaths, 9 (3.5%) strokes and 6 (2.3%) re-stenoses confirmed angiographically. One-year total survival and one-year MACCE-free survival rates according to the Kaplan-Meier analysis were 94.9% and 89.0%, showing a trend towards better outcome in the low-risk group (F-Cox=2.46; p=0.19 and F-Cox=2.17; p=0.09 respectively). CONCLUSIONS: Percutaneous transluminal angioplasty of cephalad arteries is safe and feasible, with a low periprocedural complication rate and good late outcome. Carotid artery stenting is an alternative method to CEA.

[Clinical significance of transesophageal and epiaortal echocardiography of ascending aorta in risk estimation of neurological complications in coronary artery by-pass patients]. / Znaczenie badania echokardiograficznego przezprzelykowego i epiaortalnego aorty wstepujacej w prognozowaniu ryzyka powiklan neurologicznych u chorych poddanych zabiegowi pomostowania tetnic wiencowych.

UNLABELLED: Advanced atherosclerotic changes in aortal wall are an important factor in taking decision to use minimal-invasive method of coronary artery by-pass grafting. There are some methods for diagnosing atherosclerotic changes in ascending aorta, i.e.: roentgenogram, computer tomography, magnetic resonance imaging, transthoracic echocardiography and especially transesophageal echocardiography and epiaortal echocardiography. THE AIM OF THE STUDY: To define usefulness of transesophageal and epiaortal echocardiography as a method of prognosing neurological complications in patients during coronary artery by-pass grafting. MATERIAL AND METHODS: Study group consisted of 32 consecutive patients who had coronary surgery in II Chair and Department of Cardiosurgery, Silesian Medical University in Katowice due to ischemic heart disease in whom before the surgery ascending aorta wall was evaluated with transesophageal and epiaortal echocardiography and then monitoring of microembolism was performed. Transesophageal examination was performed with Philips Sonos 7500 device with 5 MHz transducer in anesthetized patient. Ascending aorta from level of aortic valve to the aortic arch in long and short axis was evaluated. Epiaortal echocardiography was performer with 7.5 MHz vesseltransducer and Hewlett-Packard Sonos 100 CF H-P device immediately after chest opening. Transesophageal echocardiography of ascending aorta evaluated: intima-media complex thickness and presence of atherosclerotic plaques and calcifications. Microembolism monitoring was performed in 18 patients before and during surgery. Using 2 MHz transducer placed in left and right temporal region number of microembolic incidents were evaluated. RESULTS: Analysis of relationship between aortic wall thickness and microembolic signals during cardiopulmonary by-pass was performed. During cardiopulmonary by-pass microembolic signals (from 7 to 698 signals) were detected in 7 patients. During analysis number of microembolic signals was normalized to cardiopulmonary by-pass duration time. CONCLUSIONS: There was found correlation between number of microembolic signals during cardiopulmonary by-pass and thickness of aortic posterior wall in all of it's levels, using epiaortal method in short axis. This same was found using lateral wall measurements. There was no correlation between aortic wall thickness evaluated with transesophageal echocardiography and numbers of microembolic signals.

Patient- and lesion-tailored algorithm of endovascular treatment for arterial occlusive disease of extracranial arteries supplying the brain: safety of the treatment at 30-day follow-up.

INTRODUCTION: Although surgical endarterectomy remains the treatment of choice for carotid artery stenosis, stenting plays an important role as an alternative treatment modality, especially in high-risk patients. The actual safety profile associated with stenting procedures is probably better than that reported by randomized controlled trials. AIM: To assess the safety of stent implantations in extracranial arteries supplying the brain, and also to identify risk factors associated with this procedure. MATERIAL AND METHODS: This was a post hoc analysis, with 30-day follow-up. We analyzed the results of treatment of 372 patients who underwent 408 procedures, 197 such procedures in asymptomatic, and 211 in symptomatic individuals. Stenting procedures were performed using a technique and armamentarium which were tailored to the type and anatomy of lesions. RESULTS: There were 6 (1.5%) strokes, including 2 (0.5%) major strokes, 1 ipsi- and 1 contralateral, and 4 (1.0%) minor strokes. In asymptomatic patients there was 1 (0.3%) minor stroke. Transient ischemic attacks occurred in 5 (1.2%) patients. There were 2 (0.5%) non-STEMI myocardial infarctions and 2 (0.5%) non-stroke related fatalities. Risk factors of these adverse events were diabetes mellitus, lesions localized in a tortuous segment of the artery, embolic material in the filter and bilateral stenoses of carotid arteries. Additional risk factors in asymptomatic patients were renal impairment and advanced coronary artery disease; and in symptomatic patients, grade 3 arterial hypertension, dislipidemia, cigarette smoking and lesions requiring predilatation. CONCLUSIONS: Stenting procedures of extracranial arteries supplying the brain, which are tailored to the type and anatomy of lesions, seem to be relatively safe.

Protein production and secretion in an Aspergillus nidulans mutant impaired in glycosylation.

O-glycosylation has been considered a limiting factor in protein secretion in filamentous fungi. Overexpression of the yeast DPM1 gene encoding dolichylphosphate mannose synthase (DPMS) in an Aspergillus nidulans mutant (BWB26A) deficient in O-glycosylation caused an increase in the number of secretory vesicles and changes in protein secretion. However, the secretory proteins, primarily O-mannosylated glucoamylase and N-glycosylated invertase, were mainly trapped in the periplasmic space. Different glycoforms of invertase were found insite the cells, in the periplasmic space and in the cultivation medium. Our data point to the importance of the cell wall as a barrier in protein secretion.

[Composed angioplasty of the multilevel right common and internal carotid artery stenoses with implantation stents with used of proximal and distal protection system]. / Zlozona angioplastyka prawej tetnicy szyjnej wewnetrznej i wspólnej z implantacja stentów z zastosowaniem równoczesnej protekcji proksymalnej i dystalnej.

We present a case of a 73 year-old man with, long-segment thigh stenosis in the right common and internal carotid artery with occlusion left common carotid artery. Complex angioplasty with modification proximal and distal system protection was successful performed. There were no procedure-related complications. We concluded that in very complex, multilevel lesions in carotid artery, there is a place for safe double system protection. This maneuver can increase safe of the carotid artery stenting and minimise potential complications.