Nox4 oxidase overexpression specifically decreases endogenous Nox4 mRNA and inhibits angiotensin II-induced adventitial myofibroblast migration

The vascular adventitia is emerging as an important modulator of vessel remodeling. Adventitial myofibroblasts migrate to the neointima after balloon angioplasty, contributing to restenosis. We postulated that angiotensin II (Ang II) enhances adventitial myofibroblast migration in vitro via reduced nicotinamide-adenine dinucleotide phosphate oxidase-derived H(2)O(2) and that Nox4-based oxidase promotes migration. Ang II increased myofibroblast migration in a concentration-dependent manner, with a peak increase of 1023+/-83%. Rat adventitial myofibroblasts were cotransfected with human Nox4 and human p22-phox plasmids or an empty vector. PCR showed an 8-fold increase in human Nox4 and human p22-phox plasmid expression. Using RT-PCR with primers specifically designed for rat reduced nicotinamide-adenine dinucleotide phosphate oxidases, endogenous Nox levels were determined. Ang II decreased endogenous Nox4 and Nox1 mRNA to 41% and 27% of control, respectively, but had no effect on Nox2. Cotransfection with human Nox4 and human p22-phox plasmids combined with Ang II reduced endogenous Nox4 mRNA levels (37+/-5% of control; P<0.05), whereas it had no significant effect on Nox1 or Nox2. In empty vector-transfected cells, Ang II increased myofibroblast migration by 192+/-32% versus vehicle (P<0.01) while increasing H(2)O(2) (473+/-22% versus control; P<0.001). Cotransfection with human Nox4 and human p22-phox plasmids decreased Ang II-induced migration (46+/-6%; P<0.001) in parallel with attenuation of H(2)O(2) production (23+/-8% versus empty vector; P<0.05). Our data suggest that Nox4 promotes Ang II-induced myofibroblast migration via an H(2)O(2)-dependent pathway. The data also suggest that Nox4 causes feedback inhibition of its own expression in adventitial myofibroblasts.     

Photovoltaic electrocoagulation treatment of olive mill wastewater using an external-loop airlift reactor

Over the years, olive mill wastewater (OMW), which is generated from olive oil production has attracted considerable attention as one of the most polluted wastewaters due to its high concentrations of toxic compounds. It is a truism nowadays to guide research towards a novel and efficient technologies in order to treat this hazardous waste.In this paper, detoxification of OMW in an external loop airlift reactor (ALR) by electrocoagulation (EC) powered by photovoltaic solar system as a renewable and sustainable energy source was investigated. The effects of the operating parameters, such as electrolysis time, initial pH, current density (CD), and the axial position of the electrode were studied in continuous flow with two aluminum electrodes. A rate of abatement of about 79.24% for COD, 94.82% for polyphenols and 97.87% for dark color was observed within 40 min of treatment at 32.14 mA/cm² current density, a position of the electrodes at 35 cm from the bottom of the riser compartment, and initial pH of 5.6. Furthermore, under these optimal conditions, the power supplied by photovoltaic cells led to amount of electrode dissolved (ELC) and specific energy consumption (SEC) of around 0.1118 kg/m³ and 9.86 kWh/m³, respectively. It has been proven that the EC treatment of OMW in ALR coupled with renewable energy source offers a low operation cost (0.2 USD/m³) compared to other treatment processes.     

Walker 256 tumour cells increase substance P immunoreactivity locally and modify the properties of the blood–brain barrier during extravasation and brain invasion

It is not yet known how tumour cells traverse the blood–brain barrier (BBB) to form brain metastases. Substance P (SP) release is a key component of neurogenic inflammation which has been recently shown to increase the permeability of the BBB following CNS insults, making it a possible candidate as a mediator of tumour cell extravasation into the brain. This study investigated the properties of the BBB in the early stages of tumour cell invasion into the brain, and the possible involvement of SP. Male Wistar rats were injected with Walker 256 breast carcinoma cells via the internal carotid artery and euthanised at 1, 3, 6 and 9 days post tumour inoculation. Culture medium-injected animals served as controls at 1 and 9 days. Evidence of tumour cell extravasation across the BBB was first observed at 3 days post-inoculation, which corresponded with significantly increased albumin (p < 0.05) and SP immunoreactivity (p < 0.01) and significantly reduced endothelial barrier antigen labelling of microvessels when compared to culture medium control animals (p < 0.001). By day 9 after tumour cell inoculation, 100 % of animals developed large intracranial neoplasms that had significantly increased albumin in the peri-tumoral area (p < 0.001). The increased SP immunoreactivity and altered BBB properties at 3 days post-inoculation that coincided with early tumour invasion may be indicative of a mechanism for tumour cell extravasation into the brain. Thus, extravasation of tumour cells into the brain to form cerebral metastases may be a SP-mediated process.     

A new automatic image analysis method for assessing estrogen receptors’ status in breast tissue specimens

Manual assessment of estrogen receptors′ (ER) status from breast tissue microscopy images is a subjective, time consuming and error prone process. Automatic image analysis methods offer the possibility to obtain consistent, objective and rapid diagnoses of histopathology specimens. In breast cancer biopsies immunohistochemically (IHC) stained for ER, cancer cell nuclei present a large variety in their characteristics that bring various difficulties for traditional image analysis methods. In this paper, we propose a new automatic method to perform both segmentation and classification of breast cell nuclei in order to give quantitative assessment and uniform indicators of IHC staining that will help pathologists in their diagnostic. Firstly, a color geometric active contour model incorporating a spatial fuzzy clustering algorithm is proposed to detect the contours of all cell nuclei in the image. Secondly, overlapping and touching nuclei are separated using an improved watershed algorithm based on a concave vertex graph. Finally, to identify positive and negative stained nuclei, all the segmented nuclei are classified into five categories according to their staining intensity and morphological features using a trained multilayer neural network combined with Fisher's linear discriminant preprocessing. The proposed method is tested on a large dataset containing several breast tissue images with different levels of malignancy. The experimental results show high agreement between the results of the method and ground-truth from the pathologist panel. Furthermore, a comparative study versus existing techniques is presented in order to demonstrate the efficiency and the superiority of the proposed method.     

Efficacy of different doses of sugammadex after continuous infusion of rocuronium

AIM: To evaluate the effects of two different doses of sugammadex after maintenance anesthesia with sevofluorane and remifentanil and deep rocuroniuminduced neuromuscular blockade(NMB).METHODS: Patients between 20 and 65 years of age, with American Society of Anesthesiologists physical status classification Ⅰ-Ⅱ, undergoing gynecological surgery were included in a prospective, comparative and randomized study. NMB was induced with an injection of 0.6 mg/kg of rocuronium followed by continuous infusion of 0.3-0.6 mg/kg per hour to maintain a deep block. Anesthesia was maintained with sevofluorane and remifentanil. Finally, when surgery was finished, a bolus of 2 mg/kg(group A) or 4 mg/kg(group B) of sugammadex was applied when the NMB first response in the train-of-four was reached. The primary clinical endpoint was time to recovery to a train-of-four ratio of 0.9. Other variables recorded were the time until recovery of train-of-four ratio of 0.7, 0.8, hemodynamic variables(arterial blood pressure and heart rate at baseline, starting sugammadex, and minutes 2, 5 and 10) and adverse events were presented after one hour in the post-anesthesia care unit.RESULTS: Thirty-two patients were included in the study: 16 patients in group A and 16 patients in group B. Only 14 patients each group were recorded because arterial pressure values were lost in two patients from each group in minute 10. The two groups were comparable. Median recovery time from starting of sugammadex administration to a train-of-four ratio of 0.9 in group A and B was 129 and 110 s, respectively.The estimated difference in recovery time between groups was 24 s(95%CI: 0 to 45 s, Hodges-Lehmann estimator), entirely within the predefined equivalence interval. Times to recovery to train-of-four ratios of 0.8(group A: 101 s; group B: 82.5 s) and 0.7(group A: 90 s; group B: 65 s) from start of sugammadex administration were not equivalent between groups. There was not a significant variation in the arterial pressure and heart rate values between the two groups and none of the patients showed any clinical evidence of residual or recurrent NMB. CONCLUSION: A dose of 2 mg/kg of sugammadex after continuous rocuronium infusion is enough to reverse the NMB when first response in the Train-OfFour is reached.     

The Impact of COVID-19 Pandemic on the Hospital Management of TAVI Patients: TAVI Team Thoughts and Recommendation

The COVID-19 Pandemic has put enormous pressure on the healthcare system globally, causing many healthcare organizations all over the world to cancel or stop elective procedures in their cardiac catheterization laboratoires. This delay in elective procedures with no doubt has led to a suspension of patient care primarily to those with severe aortic stenosis, which might place them at higher risk for cardiovascular complications like sudden death and heart failure.Health Care Worker are faced with the uncertainty of contracting infections while performing procedures in patients with a confirmed diagnosis of COVID-19 or suspected cases. This unprecedented situation is very challenging for the safety of Health Care Worker. Hence, in this article, we aim to summarize some of the current guidelines as to how to triage patients in need for Trans Catheter Aortic Valve Implantation (TAVI), during this ongoing pandemic, and will address some necessary considerations related to the preparation of catheterization laboratories and personal during the COVID-19 pandemic.     

Novel coding,translation, and gene expression of a replicating covalently closed circular RNA of 220 nt

The highly structured (64% GC) covalently closed circular (CCC) RNA (220 nt) of the virusoid associated with rice yellow mottle virus codes for a 16-kDa highly basic protein using novel modalities for coding, translation, and gene expression. This CCC RNA is the smallest among all known viroids and virusoids and the only one that codes proteins. Its sequence possesses an internal ribosome entry site and is directly translated through two (or three) completely overlapping ORFs (shifting to a new reading frame at the end of each round). The initiation and termination codons overlap UGAUGA (underline highlights the initiation codon AUG within the combined initiation-termination sequence). Termination codons can be ignored to obtain larger read-through proteins. This circular RNA with no noncoding sequences is a unique natural supercompact “nanogenome.”Viroids and virusoids (viroid-like satellite RNAs) are typically small (220–450 nt) covalently closed circular (CCC) RNAs with no coding capacity (i.e., no genetic information) (1–3) and are the smallest replicating circular RNA pathogens (3, 4). Because of their circular nature, they usually replicate through a rolling circle model to produce larger concatemers (4, 5) which are then processed into monomeric forms with a self-splicing hammerhead ribozyme (virusoids and viroids in the Avsunviroidae family) (6, 7) or by cellular enzymes (8). We have previously reported (9) the characterization and nucleotide sequence of the smallest circular virusoid (220 nt), that of the rice yellow mottle virus (sobemovirus) (RYMV). Like other known virusoids, the small circular satellite of RYMV (scRYMV) depends on a helper virus RYMV for replication and packaging (9, 10).In silico translation of scRYMV revealed the presence of an unusual ORF capable of initiating translation from the AUG in the sequence UGAUGA of the 220-nt circular RNA by internal ribosome binding site (IRBS). As 220 is not an integer multiple of 3, after the first round of translation, the same circular sequence (or possibly the linear head-to-tail concatemers generated in vivo during rolling-circle replication) would be read in a different frame register. After the second round of translation, termination at the same initiation–termination sequence UGAUGA would result in the production of a highly basic 16-kDa protein with the N- and C-terminal halves of the protein encoded by the same 220-nt sequence but read in two distinct, totally overlapping frames. This virusoid could also suppress the leaky tandem termination codons (UGAUGA) to read the same sequence in a third frame and produce a new (18 kDa) read-through protein with an 18-aa C-terminal extension ended by a UAG codon. However, even the latter UAG termination codon may occasionally be ignored to generate longer proteins.In this report, we present evidence demonstrating that the putative ORF(s) deduced from the 220-nt circular RNA sequence as described above are indeed operational.Although this scRYMV RNA is classified as a virusoid, we report here that this virusoid is the only one so far found to encode for a protein. Modalities of initiation of translation (e.g., overlapped initiation and termination codons), the distinct N- and C-terminal halves of the 16-kDa protein translated from the same 220-nt circular sequence and the generation of read-through proteins were examined. We discuss the evolutionary implications of the genetic information and biological functions densely packed into a 220-nt nanogenome.     

Scanning electron microscopic assessment of coating irregularities and their precursors in unexpanded durable polymer‐based drug‐eluting stents

Objectives : To assess and quantify coating irregularities on unexpanded and expanded durable polymer‐based drug‐eluting stents (DES) to gain insights into the origin of coating irregularities. Background: Previous scanning electron microscopy (SEM) studies in various expanded DES revealed differences in frequency and size of coating irregularities between DES types and specific distribution patterns, however, the origin of these irregularities is unclear. Methods: We assessed at bench side a total of 1,200 SEM images obtained in 30 DES samples (15 expanded and 15 unexpanded) of Cypher Select Plus, Taxus Liberté, Endeavor, Xience V, and resolute. Results: For most coating irregularities seen on expanded DES (72%; 23/32), a matching irregularity (n = 18/23) and/or its precursor (n = 11/23) was observed in unexpanded DES. Unexpanded Cypher select showed (small) crater lesions and cracks together with precursors of “peeling.” On unexpanded Taxus Liberté, thinning of polymer, small bare metal areas, wrinkles, and one precursor type were found. Unexpanded endeavor showed cracks, small bare metal areas, crater lesions, and precursors of the latter. Unexpanded Xience V and resolute mainly revealed crater lesions and their precursors. On unexpanded versus expanded DES, there was no difference in measured frequency of coating irregularities and precursors (P = ns) with the exception of more bare metal areas on expanded Taxus Liberte (P = 0.01). Conclusions : Most coating irregularities, or the potential to develop them, are inherent to the unexpanded DES. Important determinants of the formation of coating irregularities may be the stent geometry and the physical properties of the coating, while stent‐balloon interaction plays no major role. © 2011 Wiley Periodicals, Inc.