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Background: Despite advances in surgery and pharmacotherapy, there remains significant residual ischemic risk after coronary artery bypass grafting surgery. Methods: In REDUCE-IT (Reduction of Cardiovascular Events With Icosapent Ethyl–Intervention Trial), a multicenter, placebo-controlled, double-blind trial, statin-treated patients with controlled low-density lipoprotein cholesterol and mild to moderate hypertriglyceridemia were randomized to 4 g daily of icosapent ethyl or placebo. They experienced a 25% reduction in risk of a primary efficacy end point (composite of cardiovascular death, myocardial infarction, stroke, coronary revascularization, or hospitalization for unstable angina) and a 26% reduction in risk of a key secondary efficacy end point (composite of cardiovascular death, myocardial infarction, or stroke) when compared with placebo. The current analysis reports on the subgroup of patients from the trial with a history of coronary artery bypass grafting. Results: Of the 8179 patients randomized in REDUCE-IT, a total of 1837 (22.5%) had a history of coronary artery bypass grafting, with 897 patients randomized to icosapent ethyl and 940 to placebo. Baseline characteristics were similar between treatment groups. Randomization to icosapent ethyl was associated with a significant reduction in the primary end point (hazard ratio [HR], 0.76 [95% CI, 0.63–0.92]; P =0.004), in the key secondary end point (HR, 0.69 [95% CI, 0.56–0.87]; P =0.001), and in total (first plus subsequent or recurrent) ischemic events (rate ratio, 0.64 [95% CI, 0.50–0.81]; P =0.0002) compared with placebo. This yielded an absolute risk reduction of 6.2% (95% CI, 2.3%–10.2%) in first events, with a number needed to treat of 16 (95% CI, 10–44) during a median follow-up time of 4.8 years. Safety findings were similar to the overall study: beyond an increased rate of atrial fibrillation/flutter requiring hospitalization for at least 24 hours (5.0% vs 3.1%; P =0.03) and a nonsignificant increase in bleeding, occurrences of adverse events were comparable between groups. Conclusions: In REDUCE-IT patients with a history of coronary artery bypass grafting, treatment with icosapent ethyl was associated with significant reductions in first and recurrent ischemic events. Registration: URL: https://www.clinicaltrials.gov ; Unique identifier: NCT01492361.

BACKGROUND: To investigate the association between altered sex hormone expression and long-term right ventricular (RV) adaptation and progression of right heart failure in a Dutch cohort of Pulmonary Arterial Hypertension (PAH)-patients across a wide range of ages. METHODS: In this study we included 279 PAH-patients, of which 169 females and 110 males. From 59 patients and 21 controls we collected plasma samples for sex hormone analysis. Right heart catheterization (RHC) and/or cardiac magnetic resonance (CMR) imaging was performed at baseline. For longitudinal data analysis, we selected patients that underwent a RHC and/or CMR maximally 1.5 years prior to an event (death or transplantation, N = 49). RESULTS: Dehydroepiandrosterone-sulfate (DHEA-S) levels were reduced in male and female PAHpatients compared to controls, whereas androstenedione and testosterone were only reduced in female patients. Interestingly, low DHEA-S and high testosterone levels were correlated to worse RV function in male patients only. Subsequently, we analyzed prognosis and RV adaptation in females stratified by age. Females < 45years had best prognosis in comparison to females & GE;55years and males. No differences in RV function at baseline were observed, despite higher pressure-overload in females < 45years. Longitudinal data demonstrated a clear distinction in RV adaptation. Although females < 45years had an event at a later time point, RV function was more impaired at end-stage disease. CONCLUSIONS: Sex hormones are differently associated with RV function in male and female PAHpatients. DHEA-S appeared to be lower in male and female PAH-patients. Females < 45years could persevere pressure-overload for a longer time, but had a more severe RV phenotype at end-stage disease.J Heart Lung Transplant 2022;41:445-457 (c) 2021 The Author(s). Published by Elsevier Inc. on behalf of International Society for Heart and Lung Transplantation. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)

The objective: to investigate the influence of the scheme of basic pharmacologic therapy on the dynamics of carbohydrate and lipid metabolism in a group of patients with type 2 diabetes on the background of the use of group DSME. Materials and methods. The study was performed on the basis of the Department of Family Medicine and Outpatient Care within the territorial boundaries of the Kyiv agglomeration. The study included 117 patients with type 2 diabetes. The study group of patients was randomly divided into a groups of classroom training (38 people), a group of remote training (39 people) and a control group (40 people), which did not receive additional training load. Subsequently, patients from the study groups took the appropriate course in classroom (outpatient) or remote (using Skype video conferencing) formats. The education of patients methodically and meaningfully complied with the consensus recommendations of the American Diabetes Association, the American Association of Diabetes Educators and the Academy of Nutrition and Dietetics. Subsequently, the relevant indicators of diabetes were monitored at 3 and 12 months after the start of the study and the correlation of their dynamics with the basic scheme of pharmacologic therapy ts and its change during the observation period was analyzed. Results. During the observation period, changes in the scheme of hypoglycemic and hypolipidemic therapy were made in 10 (8,5 %) and 20 (17,0 %) patients. It should be noted that the relevant changes were bilateral, both in the direction of simplification and in the direction of intensification of drug therapy regimens. Analysis of the ratio of 3- and 12-month dynamics of carbohydrate and lipid metabolism and the scheme of hypoglycemic and hypolipidemic therapy in all study groups did not reveal a significant correlation. Conclusions. There is no significant correlation between the basic scheme of hypoglycemic and hypolipidemic therapy with the dynamics of carbohydrate and lipid metabolism at the stages of 3- and 12-month check-points. The lack of influence of the pharmacologic therapy scheme on the positive dynamics of diabetes mellitus emphasizes the importance of group DSME in patients with type 2 diabetes mellitus as a separate independent factor of improving of the disease.

The article is devoted to the issue of legal regulation of medical services in Ukraine. The main focus is on family medicine and regulation of the relationship between the family doctor and the patient with whom the declaration was signed, as well as the topic of reforming the health care system of Ukraine, which began in 2015. After all, it was the reform of the health care system that opened such a state institution for Ukraine as family medicine. The relevance of this research topic is that the quality of medical services is one of the main indicators of the effective functioning of the health care system. It is the factor of proper provision of medical services that contributes to: population size, mortality, life expectancy and, in general, public health and population demographics. These important factors determine the development of the state and its socioeconomic situation. If we do not take into account the modern fundamental factor of quality, the medical system will not be able to function properly, and as a result will have problems and shortcomings that will prevent the country from moving forward, both in medicine and in other areas. After the implementation of the first stage of reform, the Ukrainian health care system has undergone significant changes. The main of which was family medicine and all its principles and innovations. The topic of family medicine in Ukraine is quite important, because the family doctor is almost the first to whom the patient turns, so attention should be focused on primary care in order for this institute of medicine to function well and efficiently. Today, Ukraine faces a problem such as the COVID-19 pandemic, which has led to certain gaps in health care, such as inefficient and poor quality of primary care services, lack of responsibility for quality on the part of both doctors and patients, and society. dissatisfied with the reform of the health care system in Ukraine, and the lack of legal force in declarations between family doctors and patients is also a problem today. These problems were not so obvious and noticeable before the COVID-19 epidemic, but it is this situation with the coronavirus that highlights all the above gaps.

Purpose. To evaluate the hypotensive effect of endotrabeculectomy as a standalone operation and in combination with cataract phacoemulsification in patients with primary open-angle glaucoma. Methods. There were 88 patients (88 eyes) with open-angle glaucoma under our observation. The patients of the first group (23 patients, 23 eyes) underwent standalone endotrabeculectomy. The patients of the second group (65 patients, 65 eyes) underwent endotrabeculectomy in combination with cataract phacoemulsification and intraocular lens implantation. Results. When comparing the IOP of the two groups, we found that no significant difference in its decrease depending on the type of surgery was detected, in all cases p > 0.05. A significant difference was found only when comparing the “Before surgery” subgroups – p = 0.031. When comparing the usage of glaucoma medications, we found that at the first 6 months, no significant difference in quantities of drugs depending on the type of surgery was found (p > 0.05), but in the next 6 months, there were significantly higher amounts of glaucoma medications used in the frst group to reach the target IOP than in the second group, which can be explained by the slightly lower baseline IOP and the lower number of glaucoma medications instilled during the preoperative period in the second group. Conclusion. Endotrabeculectomy is an effective operation for patients with primary open-angle glaucoma, both as an stand-alone surgery and in combination with cataract phacoemulsification.

Experiments were undertaken to identify the nature of a previously identified inhibitor of PAF-induced platelet aggregation (PA) in human saliva. Human saliva fractionated by preparative thin layer chromatography (TLC) yielded a fraction that co-migrated with fatty acids (FAs) and inhibited PAF-induced aggregation of platelets. Synthetic FAs tested for their capacities to inhibit 0.1 nM PAF-induced PA showed that only the cis-unsaturated compounds were inhibitory with activities of some of the polyunsaturated FAs (PUFA) reaching almost 100% at 20 μM. Eicosapentanoic acid (EPA) and 8,11,14-eicosatrienoic acid also deaggregated the PAF-induced aggregates. With the exception of oleic acid (OLA), cis-monounsaturated FAs, and elaidic acid, the trans isomer of OLA, were poor inhibitors. In a direct comparison with other platelet agonists, ADP, thrombin, and ionophore A23187, the active saliva fraction and selected individual FAs inhibited, to greater or lesser extent, PA induced by each of the agonists. EPA, OLA, linoleic acid (LNA), and the active saliva fraction were potent inhibitors of ADP-induced PA, EPA completely inhibited thrombin-induced PA and the saliva fraction showed only weak - moderate inhibitory activity to both thrombin- and ionophore A23187-induced PA. Other reports of endogenous PAF inhibitors in mammalian tissues are compared to the present results. PAF can trigger and amplify inflammatory cascades suggesting a possible modulation role for cis-unsaturated FAs in some diseases.

FZR1, which encodes the Cdh1 subunit of the Anaphase Promoting Complex, plays an important role in neurodevelopment by regulating cell cycle and by its multiple post-mitotic functions in neurons. In this study, evaluation of 250 unrelated patients with developmental epileptic encephalopathies (DEE) and a connection on GeneMatcher led to the identification of three de novo missense variants in FZR1. Two variants led to the same amino acid change. All individuals had a DEE with childhood onset generalized epilepsy, intellectual disability, mild ataxia and normal head circumference. Two individuals were diagnosed with the DEE subtype Myoclonic Atonic Epilepsy (MAE). We provide gene burden testing using two independent statistical tests to support FZR1 association with DEE. Further, we provide functional evidence that the missense variants are loss-of-function (LOF) alleles using Drosophila neurodevelopment assays. Using three fly mutant alleles of the Drosophila homolog fzr and overexpression studies, we show that patient variants do not support proper neurodevelopment. With the recent report of a patient with neonatal-onset DEE with microcephaly who also carries a de novo FZR1 missense variant, our study consolidates the relationship between FZR1 and DEE, and expands the associated phenotype. We conclude that heterozygous LOF of FZR1 leads to DEE associated with a spectrum of neonatal to childhood onset seizure types, developmental delay and mild ataxia. Microcephaly can be present but is not an essential feature of FZR1-encephalopathy. In summary, our approach of targeted sequencing using novel gene candidates and functional testing in Drosophila will help solve undiagnosed MAE/DEE cases.

FZR1, which encodes the Cdh1 subunit of the anaphase-promoting complex, plays an important role in neurodevelopment by regulating the cell cycle and by its multiple post-mitotic functions in neurons. In this study, evaluation of 250 unrelated patients with developmental and epileptic encephalopathies and a connection on GeneMatcher led to the identification of three de novo missense variants in FZR1. Whole-exome sequencing in 39 patient–parent trios and subsequent targeted sequencing in an additional cohort of 211 patients was performed to identify novel genes involved in developmental and epileptic encephalopathy. Functional studies in Drosophila were performed using three different mutant alleles of the Drosophila homologue of FZR1 fzr. All three individuals carrying de novo variants in FZR1 had childhood-onset generalized epilepsy, intellectual disability, mild ataxia and normal head circumference. Two individuals were diagnosed with the developmental and epileptic encephalopathy subtype myoclonic atonic epilepsy. We provide genetic-association testing using two independent statistical tests to support FZR1 association with developmental and epileptic encephalopathies. Further, we provide functional evidence that the missense variants are loss-of-function alleles using Drosophila neurodevelopment assays. Using three fly mutant alleles of the Drosophila homologue fzr and overexpression studies, we show that patient variants can affect proper neurodevelopment. With the recent report of a patient with neonatal-onset with microcephaly who also carries a de novo FZR1 missense variant, our study consolidates the relationship between FZR1 and developmental and epileptic encephalopathy and expands the associated phenotype. We conclude that heterozygous loss-of-function of FZR1 leads to developmental and epileptic encephalopathies associated with a spectrum of neonatal to childhood-onset seizure types, developmental delay and mild ataxia. Microcephaly can be present but is not an essential feature of FZR1-encephalopathy. In summary, our approach of targeted sequencing using novel gene candidates and functional testing in Drosophila will help solve undiagnosed myoclonic atonic epilepsy or developmental and epileptic encephalopathy cases.

The stalled fork protection pathway mediated by breast cancer 1/2 (BRCA1/2) proteins is critical for replication fork stability. However, it is unclear whether additional mechanisms are required to maintain replication fork stability. We describe a hitherto unknown mechanism, by which the SWI/SNF-related matrix-associated actindependent regulator of chromatin subfamily-A containing DEAD/H box-1 (SMARCAD1) stabilizes active replication forks, that is essential to maintaining resistance towards replication poisons. We find that SMARCAD1 prevents accumulation of 53BP1-associated nucleosomes to preclude toxic enrichment of 53BP1 at the forks. In the absence of SMARCAD1, 53BP1 mediates untimely dissociation of PCNA via the PCNA-unloader ATAD5, causing frequent fork stalling, inefficient fork restart, and accumulation of single-stranded DNA. Although loss of 53BP1 in SMARCAD1 mutants rescues these defects and restores genome stability, this rescued stabilization also requires BRCA1-mediated fork protection. Notably, fork protection-challenged BRCA1-deficient naïve- or chemoresistant tumors require SMARCAD1-mediated active fork stabilization to maintain unperturbed fork progression and cellular proliferation.

Aim. To determine the influence of the polymorphic variants of the base (OGG1 and XRCC1) and nucleotide (ERRC2/XPD and ERRC6/CSB) excision repair genes on the mutational and epigenetic status of bladder tumors. Methods. For this study, we used previously obtained data on the polymorphism of DNA repair genes in bladder cancer (BC) patients, whose tumor tissue samples were analyzed for the presence of key mutational and epigenetic changes. Results. Genotypes containing at least one minor OGG1 rs1052133 allele were significantly associated with an increased frequency of RAS family gene mutations and a reduced frequency of PIK3CA mutations. The polymorphisms of both base excision repair genes influenced the epigenetic variability of urothelial carcinomas, the modifying effect of OGG1 rs1052133 manifesting in ISL1 methylation and XRCC1 rs25487 impacting p16 or TIMP3 methylation. The minor ERRC6/CSB rs2228526 allele was associated with RAS mutations and lack of TIMP3 methylation. Likewise, carriers of the genotypes containing at least one minor ERRC2/XPD rs1799793 allele were less frequently found to have methylated RUNX3 gene in tumor tissues. Conclusions. The polymorphisms of the base (OGG1, XRCC1) and nucleotide (ERRC2/XPD, ERRC6/CSB) excision repair genes modify the mutational and epigenetic variability of a number of key BC genes. The study of the mechanisms of such interactions is necessary for understanding the molecular basis of BC pathogenesis. Keywords: bladder cancer, OGG1, XRCC1, ERRC2/XPD, ERRC6/CSB gene polymorphism, mutation, methylation.

Many chemotherapy drugs block tumor cell division by damaging DNA. DNA polymerases eta (Pol η), iota (Pol ι), kappa (Pol κ), REV1 of the Y-family and zeta (Pol ζ) of the B-family efficiently incorporate nucleotides opposite a number of DNA lesions during translesion DNA synthesis. Primase-polymerase PrimPol and the Pol α-primase complex reinitiate DNA synthesis downstream of the damaged sites using their DNA primase activity. These enzymes can decrease the efficacy of chemotherapy drugs, contribute to the survival of tumor cells and to the progression of malignant diseases. DNA polymerases are promising targets for increasing the effectiveness of chemotherapy, and mutations and polymorphisms in some DNA polymerases can serve as additional prognostic markers in a number of oncological disorders.

Y-family DNA polymerase iota (Pol ι) is involved in DNA damage response and tolerance. Mutations and altered expression level of POLI gene are linked to a higher incidence of cancer. We biochemically characterized five active site polymorphic variants of human Pol ι: R71G (rs3218778), P118L (rs554252419), I236M (rs3218784), E251K (rs3218783) and P365R (rs200852409). We analyzed fidelity of nucleotide incorporation on undamaged DNA, efficiency and accuracy of DNA damage bypass, as well as 5′-deoxyribophosphate lyase (dRP-lyase) activity. The I236M and P118L variants were indistinguishable from the wild-type Pol ι in activity. The E251K and P365R substitutions altered the spectrum of nucleotide incorporation opposite several undamaged DNA bases. The P365R variant also reduced the dRP-lyase activity and possessed the decreased TLS activity opposite 8-oxo-G. The R71G mutation dramatically affected the catalytic activities of Pol ι. The reduced DNA polymerase activity of the R71G variant correlated with an enhanced fidelity of nucleotide incorporation on undamaged DNA, altered lesion-bypass activity and reduced dRP-lyase activity. Therefore, this amino acid substitution likely alters Pol ι functions in vivo.

Percutaneous coronary intervention (PCI) is typically performed with image guidance using X-ray angiograms in which coronary arteries are opacified with X-ray opaque contrast agents. Interventional cardiologists typically navigate instruments using non-contrast-enhanced fluoroscopic images, since higher use of contrast agents increases the risk of kidney failure. When using fluoroscopic images, the interventional cardiologist needs to rely on a mental anatomical reconstruction. This paper reports on the development of a novel dynamic coronary roadmapping approach for improving visual feedback and reducing contrast use during PCI. The approach compensates cardiac and respiratory induced vessel motion by ECG alignment and catheter tip tracking in X-ray fluoroscopy, respectively. In particular, for accurate and robust tracking of the catheter tip, we proposed a new deep learning based Bayesian filtering method that integrates the detection outcome of a convolutional neural network and the motion estimation between frames using a particle filtering framework. The proposed roadmapping and tracking approaches were validated on clinical X-ray images, achieving accurate performance on both catheter tip tracking and dynamic coronary roadmapping experiments. In addition, our approach runs in real-time on a computer with a single GPU and has the potential to be integrated into the clinical workflow of PCI procedures, providing cardiologists with visual guidance during interventions without the need of extra use of contrast agent.

The transcription factor Zeb2 controls fate specification and subsequent differentiation and maturation of multiple cell types in various embryonic tissues. It binds many protein partners, including activated Smad proteins and the NuRD co-repressor complex. How Zeb2 subdomains support cell differentiation in various contexts has remained elusive. Here, we have studied the role of Zeb2 and its domains in neurogenesis and neural differentiation in the young postnatal ventricular-subventricular zone (V-SVZ), where neural stem cells generate olfactory bulb-destined interneurons. Conditional Zeb2 knockouts and separate acute loss- and gain-of-function approaches indicated that Zeb2 is essential to control apoptosis and neuronal differentiation of V-SVZ progenitors before and after birth, and identified Sox6 as Zeb2-dependent and potential downstream target gene. Zeb2 genetic inactivation impaired the differentiation potential of the V-SVZ niche in a cell-autonomous fashion. We also provide evidence that its normal function in the V-SVZ involves non-autonomous mechanisms as well. Additionally, we could demonstrate distinct roles for Zeb2 protein-binding domains, suggesting that Zeb2 partners co-determine neuronal output from the mouse V-SVZ in both quantitative and qualitative manners in early postnatal life. ispartof: Development vol:147 issue:10 ispartof: location:England status: published

Promoter hypermethylation of tumor suppressor genes is one of the mechanisms of epigenetic regulation disturbance of gene expression and is often observed in different cancer types. The profile of mutational and epigenetic changes characterizes a malignant potential of a tumor, as well as its ability to invade and metastasize.The aim of the study was to determine a prognostic value of p16, p14ARF and TIMP3 gene methylation in the group of 158 bladder cancer patients. Epigenetic changes in these genes were observed with a frequency of 11.4, 0 and 10.8 %, respectively, and did not depend on clinic-morphological characteristics.A statistically significant association of p16 and TIMP3 abnormal methylation with smoking was found, indicating a possible influence of tobacco smoke carcinogens on the occurrence of these epigenetic changes. In the multivariate Cox regression analysis, p16 promoter hypermethylation was an independent predictor for bladder cancer progression (HR 6.84; 95 % CI 1.6–29.9; р = 0.011).The use of the data on the p16 methylation status may improve the accuracy of prognosis of the bladder cancer clinical course and the selection of appropriate treatment strategy.

To accurately predict the tumor behavior and individualize the treatment approach, new methods for bladder cancer (BC) prognosis are required. The most promising prognostic markers are the mutational and epigenetic changes of genes involved in maintaining cellular homeostasis. In the present study, we evaluated the influence of p16 promoter hypermethylation on the risk of recurrence, progression and disease outcome in the group of 158 BC patients. p16 epigenetic changes were found in 11.4 % of urothelial carcinomas and did not depend on clinicоmorphological characteristics. However, in the subgroup of patients with non-muscle invasive tumors, p16 abnormal methylation was significantly associated with smoking, and in the subgroup of patients with muscle-invasive BC, it was linked to a high tumor grade (G3). In the multivariate Cox regression analysis, p16 promoter hypermethylation was an independent predictor for bladder cancer progression (HR 6.84; 95 % CI 1.6–29.9; р = 0.011). The use of the data on the p16 methylation status may improve the accuracy of prognosis of the bladder cancer clinical course and the selection of appropriate treatment strategy.

W artykule przedstawiono podstawy zarządzania ryzykiem, a zwłaszcza kwestie związane z metodyką oceny ryzyka zawodowego na wybranych stanowiskach pracy. Na potrzeby oceny ryzyka zawodowego w badanej instytucji zaproponowano metodę Risk Score oraz przedstawiono efekty tej oceny na przykładzie wybranego stanowiska z użyciem karty oceny ryzyka zawodowego, która została dostosowana do lokalnych uwarunkowań. Przygotowana propozycja karty oceny ryzyka zawiera charakterystykę danego stanowiska pracy, identyfikację zagrożeń, ich źródeł, możliwych skutków i stosowanych zabezpieczeń na wybranym stanowisku oraz kartę zapoznania pracownika z oceną ryzyka zawodowego. Głównym założeniem dokonanej oceny ryzyka zawodowego było zbadanie możliwych zagrożeń, które występują w środowisku pracowniczym oraz możliwie najlepsze dostosowanie zabezpieczeń zapobiegających wystąpieniu niepożądanego zdarzenia.

Infection during pregnancy is common and the developing fetal brain is vulnerable to vertical transmission due to immaturity of the fetal immune system. Infection is a major cause of multiple organ abnormalities, including the neuraxis, due to the neurotropism of the infectious agents. This review sets out to give an overview of fetal infection, review the general principles of the nature and timing of the infectious insult with respect to outcomes, review the neuroimaging of infection by ultrasound and magnetic resonance imaging (MRI), and review the various pathogens involved, including the two most common, cytomegalovirus (CMV) and Toxoplasma, and also other common viral and nonviral infections.

In minimal invasive image guided catheterization procedures, physicians require information of the catheter position with respect to the patient’s vasculature. However, in fluoroscopic images, visualization of the vasculature requires toxic contrast agent. Static vasculature roadmapping, which can reduce the usage of iodine contrast, is hampered by the breathing motion in abdominal catheterization. In this paper, we propose a method to track the catheter tip inside the patient’s 3D vessel tree using intra-operative single-plane 2D X-ray image sequences and a peri-operative 3D rotational angiography (3DRA). The method is based on a hidden Markov model (HMM) where states of the model are the possible positions of the catheter tip inside the 3D vessel tree. The transitions from state to state model the probabilities for the catheter tip to move from one position to another. The HMM is updated following the observation scores, based on the registration between the 2D catheter centerline extracted from the 2D X-ray image, and the 2D projection of 3D vessel tree centerline extracted from the 3DRA. The method is extensively evaluated on simulated and clinical datasets acquired during liver abdominal catheterization. The evaluations show a median 3D tip tracking error of 2.3 mm with optimal settings in simulated data. The registered vessels close to the tip have a median distance error of 4.7 mm with angiographic data and optimal settings. Such accuracy is sufficient to help the physicians with an up-to-date roadmapping. The method tracks in real-time the catheter tip and enables roadmapping during catheterization procedures.

The rapid development of nanotechnologies and increased production and use of nanomaterials raise concerns about their potential toxic effects for human health and environment. To evaluate the biological effects of nanomaterials, a set of reliable and reproducible methods and development of standard operating procedures (SOPs) is required. In the framework of the European FP7 NanoValid project, three different cell viability assays (MTS, ATP content, and caspase-3/7 activity) with different readouts (absorbance, luminescence and fluorescence) and two immune assays (ELISA of pro-inflammatory cytokines IL1-β and TNF-α) were evaluated by inter-laboratory comparison. The aim was to determine the suitability and reliability of these assays for nanosafety assessment. Studies on silver and copper oxide nanoparticles (NPs) were performed, and SOPs for particle handling, cell culture, and in vitro assays were established or adapted. These SOPs give precise descriptions of assay procedures, cell culture/seeding conditions, NPs/positive control preparation and dilutions, experimental well plate preparation, and evaluation of NPs interference. The following conclusions can be highlighted from the pan-European inter-laboratory studies: Testing of NPs interference with the toxicity assays should always be conducted. Interference tests should be designed as close as possible to the cell exposure conditions. ATP and MTS assays gave consistent toxicity results with low inter-laboratory variability using Ag and CuO NPs and different cell lines and therefore, could be recommended for further validation and standardization. High inter-laboratory variability was observed for Caspase 3/7 assay and ELISA for IL1-β and TNF-α measurements., by Superb K. Misra et al.

AIM: The intensity of the most appropriate exercise to use in depressed youth is unclear due to differences in methodology and the lack of evidence documenting the effect of physical activity in children. Therefore, the authors of this study attempted to document the effectiveness of different training intensities to reduce symptoms of depression and anxiety in pre-teens. METHODS: The study included twenty-seven, randomly selected pre-adolescents (aged between 9-11 years of age) all of whom had Primary education. The participants were enrolled and, over a 5-week period, were subject to either intensive or low-to-moderate exercise programs four times a week. Psychological self-reports, as well as physical examinations, were conducted before and after such programs in blinded assessments. Psychological effects were considered the primary outcome, whilst physical condition was secondary. RESULTS: Four subjects were lost and twenty-three were analyzed. General linear model with 2 criteria revealed significant changes (p = 0.05) in trait anxiety symptoms over time in the low-to-moderate intensity group (LMIG). Within group changes followed a significant decrease in levels of anxiety (38.82 ± 2.20 to 33.36 ± 2.83, p = 0.004) and depression (10.36 ± 2.83 to 6.73 ± 1.88, p = 0.006) related symptoms amongst those in the LMIG. INTERPRETATION: This study indicated that depression and anxiety symptoms were reduced amongst a non-clinical sample of Primary educated pre-adolescents when they were subject to a low-to-moderate exercise program. The program focused on associating movement with pleasure, encouraged positive and non-competitive interactions between participants. CLINICAL TRIAL REGISTRATION: www.ClinicalTrials.gov, identifier NCT02970825, autumn 2016, updated May 7, 2018 (https://clinicaltrials.gov/ct2/show/NCT02970825).

Diagnosis of patients suffering from small-fiber neuropathy is a challenging task and requires accurate measurement of the density of nerve fibers crossing the dermal-epidermal junction in the skin. Currently this is typically done by expert manual counting in microscopy images of sliced and stained skin biopsies. It is a rather subjective and labor-intensive process that would benefit greatly from more automated approaches. Previously we have explored classical image processing methods for this, with very limited success. Here we explore the potential of convolutional neural networks and deep learning for the task. The results of preliminary experiments show the networks perform close to the expert and outperform novices and our previous method.

// Eliza Starczewska 1 , Maxime Beyaert 1 , Lucienne Michaux 2 , Marie-Christiane Vekemans 2 , Pascale Saussoy 3 , Vanesa Bol 4 , Ainhoa Arana Echarri 1 , Caroline Smal 1 , Eric Van Den Neste 1, 2 , Francoise Bontemps 1 1 de Duve Institute, Universite Catholique de Louvain, B-1200 Brussels, Belgium 2 Department of Hematology, Cliniques Universitaires Saint-Luc, Universite Catholique de Louvain, B-1200 Brussels, Belgium 3 Service de Biologie Clinique, Cliniques Universitaires Saint-Luc, Universite Catholique de Louvain, B-1200 Brussels, Belgium 4 Center for Molecular Imaging, Radiotherapy and Oncology, Institut de Recherche Experimentale et Clinique (IREC), Universite Catholique de Louvain, B-1200 Brussels, Belgium Correspondence to: Francoise Bontemps, email: francoise.bontemps@uclouvain.be Keywords: chronic lymphocytic leukemia, DNA damage, fludarabine, aphidicolin, γH2AX Received: March 09, 2016 Accepted: May 04, 2016 Published: May 20, 2016 ABSTRACT Purine analogs are among the most effective chemotherapeutic drugs for the treatment of chronic lymphocytic leukemia (CLL). However, chemoresistance and toxicity limit their clinical use. Here, we report that the DNA polymerase inhibitor aphidicolin, which displayed negligible cytotoxicity as a single agent in primary CLL cells, markedly synergizes with fludarabine and cladribine via enhanced apoptosis. Importantly, synergy was recorded regardless of CLL prognostic markers. At the molecular level, aphidicolin enhanced purine analog-induced phosphorylation of p53 and accumulation of γH2AX, consistent with increase in DNA damage. In addition, aphidicolin delayed γH2AX disappearance that arises after removal of purine analogs, suggesting that aphidicolin causes an increase in DNA damage by impeding DNA damage repair. Similarly, aphidicolin inhibited UV-induced DNA repair known to occur primarily through the nucleotide excision repair (NER) pathway. Finally, we showed that fludarabine induced nuclear import of XPA, an indispensable factor for NER, and that XPA silencing sensitized cell lines to undergo apoptosis in response to fludarabine. Together, our data indicate that aphidicolin potentiates the cytotoxicity of purine analogs by inhibiting a DNA repair pathway that involves DNA polymerases, most likely NER, and provide a rationale for manipulating it to therapeutic advantage.

Проведено проспективное исследование по оценке прогностического значения мутационного статуса гена FGFR 3 у пациентов с раком мочевого пузыря без мышечной инвазии (РМПБМИ). В исследование включено 265 пациентов, у 168 (63,4%) обнаружены мутации гена FGFR 3 . Установлено, что частота мутаций гена FGFR 3 была статистически значимо выше в высокодифференцированных опухолях (р=0,00004). При медиане наблюдения 34 мес. не выявлено статистически значимого показателя относительного риска прогрессирования РМПБМИ при наличии мутации гена FGFR 3 по сравнению с ее отсутствием (0,50; 95%ДИ 0,17-1,49; P = 0,21). При анализе прогностического значения мутационной изменчивости гена FGFR 3 в различных подгруппах было обнаружено, что у пациентов с опухолями T1 high grade (n=41) мутации гена FGFR 3 были связаны со статистически значимо лучшим прогнозом: 3-летняя выживаемость до прогрессирования при наличии мутации (n=17) составила 100% по сравнению с 71,2% (95%ДИ 42,8-99,6%) при отсутствии мутации (n=24). При остальных категориях опухолей (Ta, T1 low grade) статистически значимых различий в выживаемости до прогрессирования в зависимости от мутационного статуса FGFR 3 выявлено не было.

A novel mechanism is described for the agrin-mediated focal delivery of acetylcholine receptors (AChRs) to the postsynaptic membrane of the neuromuscular junction. Microtubule capture mediated by CLASP2 and its interaction partner, LL5β, and an intact subsynaptic actin cytoskeleton are both required for focal AChR transport to the synaptic membrane., A hallmark of the neuromuscular junction (NMJ) is the high density of acetylcholine receptors (AChRs) in the postsynaptic muscle membrane. The postsynaptic apparatus of the NMJ is organized by agrin secreted from motor neurons. The mechanisms that underlie the focal delivery of AChRs to the adult NMJ are not yet understood in detail. We previously showed that microtubule (MT) capture by the plus end–tracking protein CLASP2 regulates AChR density at agrin-induced AChR clusters in cultured myotubes via PI3 kinase acting through GSK3β. Here we show that knockdown of the CLASP2-interaction partner LL5β by RNAi and forced expression of a CLASP2 fragment blocking the CLASP2/LL5β interaction inhibit microtubule capture. The same treatments impair focal vesicle delivery to the clusters. Consistent with these findings, knockdown of LL5β at the NMJ in vivo reduces the density and insertion of AChRs into the postsynaptic membrane. MT capture and focal vesicle delivery to agrin-induced AChR clusters are also inhibited by microtubule- and actin-depolymerizing drugs, invoking both cytoskeletal systems in MT capture and in the fusion of AChR vesicles with the cluster membrane. Combined our data identify a transport system, organized by agrin through PI3 kinase, GSK3β, CLASP2, and LL5β, for precise delivery of AChR vesicles from the subsynaptic nuclei to the overlying synaptic membrane.

Dengue virus nonstructural protein 3 (NS3) is a multifunctional protein formed by a superfamily-2 RNA helicase linked to a protease domain. In this work, we report results from in vitro experiments designed to determine the oligomeric state of dengue virus NS3 helicase (NS3h) and to characterize fundamental properties of the interaction with single-stranded (ss)RNA. Pulsed field gradient-NMR spectroscopy was used to determine the effective hydrodynamic radius of NS3h, which was constant over a wide range of protein concentrations in the absence and presence of ssRNA. Size exclusion chromatography-static light scattering experiments showed that NS3h eluted as a monomeric molecule even in the presence of ssRNA. Binding of NS3h to ssRNA was studied by quantitative fluorescence titrations using fluorescein-labeled and unlabeled ssRNA oligonucleotides of different lengths, and the effect of the fluorescein label on the interaction parameters was also analyzed. Experimental results were well described by a statistical thermodynamic model based on the theory of non-specific interactions of large ligands to a one-dimensional lattice. We found that binding of NS3h to ssRNA oligonucleotides and to poly(A) is characterized by minimum and occluded binding site sizes both of 10 nucleotides and by a weak positive cooperativity between adjacent proteins. Fil: Gebhard, Leopoldo German. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina Fil: Incicco, Juan Jeremías. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina Fil: Smal, Clara. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina Fil: Gallo, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina Fil: Gamarnik, Andrea Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina Fil: Kaufman, Sergio Benjamín. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica; Argentina

Interactions with receptors for the Fc region of IgG (Fc gamma Rs) have been shown to contribute to the in vivo protection against influenza A viruses provided by broadly neutralizing antibodies (bnAbs) that bind to the viral hemagglutinin (HA) stem. In particular, Fc-mediated antibody-dependent cellular cytotoxicity (ADCC) has been shown to contribute to protection by stem-binding bnAbs. Fc-mediated effector functions appear not to contribute to protection provided by strain-specific HA head-binding antibodies. We used a panel of anti-stem and anti-head influenza A and B monoclonal antibodies with identical human IgG1 Fc domains and investigated their ability to mediate ADCC-associated Fc gamma RIIIa activation. Antibodies which do not interfere with sialic acid binding of HA can mediate Fc gamma RIIIa activation. However, the Fc gamma RIIIa activation was inhibited when a mutant HA, unable to bind sialic acids, was used. Antibodies which block sialic acid receptor interactions of HA interfered with Fc gamma RIIIa activation. The inhibition of Fc gamma RIIIa activation by HA head-binding and sialic acid receptor-blocking antibodies was confirmed in plasma samples of H5N1 vaccinated human subjects. Together, these results suggest that in addition to Fc-Fc gamma R binding, interactions between HA and sialic acids on immune cells are required for optimal Fc-mediated effector functions by anti-HA antibodies

A potent anti-vascular endothelial growth factor (VEGF) biologic and a compatible delivery system were co-evaluated for protection against wet age-related macular degeneration (AMD) over a 6month period following a single intravitreal (IVT) injection. The anti-VEGF molecule is dimeric, containing two different anti-VEGF domain antibodies (dAb) attached to a human IgG1 Fc region: a dual dAb. The delivery system is based on microparticles of PolyActive™ hydrogel co-polymer. The molecule was evaluated both in vitro for potency against VEGF and in ocular VEGF-driven efficacy models in vivo. The dual dAb is highly potent, showing a lower IC50 than aflibercept in VEGF receptor binding assays (RBAs) and retaining activity upon release from microparticles over 12months in vitro. Microparticles released functional dual dAb in rabbit and primate eyes over 6months at sufficient levels to protect Cynomolgus against laser-induced grade IV choroidal neovascularisation (CNV). This demonstrates proof of concept for delivery of an anti-VEGF molecule within a sustained-release system, showing protection in a pre-clinical primate model of wet AMD over 6months. Polymer breakdown and movement of microparticles in the eye may limit development of particle-based approaches for sustained release after IVT injection.

Item does not contain fulltext We present the case of a 2-year-old asthmatic boy with atrioventricular (AV)-reentry tachycardia following albuterol inhalation, who was later diagnosed with Wolff-Parkinson-White (WPW) syndrome. The Naranjo adverse drug reaction probability scale score for this adverse event was 7, indicating that the association between his AV-reentry tachycardia and inhalation of albuterol is probable. To our knowledge, this is the first case report that shows the potential arrhythmogenic effects of albuterol in a child with WPW syndrome. We urge clinicians to be aware of this potentially life-threatening adverse effect and to closely monitor these patients when they need beta-adrenergic drugs in case of emergency. Furthermore, this report highlights the dilemma regarding the safe treatment of pediatric patients with both asthma and WPW syndrome.

Congenital infiltrating lipomatosis of the face (CILF) is a rare disorder in which overgrowth of fatty tissue with infiltration of underlying structures leads to craniofacial deformities and asymmetry. Imaging plays an important role in the diagnosis, surgical planning and evaluation of underlying structures. To our knowledge, our case is the first to describe CILF associated with isolated enlargement of a cerebellar hemisphere in the absence of an enlarged ipsilateral cerebral hemisphere (hemimegalencephaly).

Williams-Beuren syndrome (WBS) is the chromosomal disorder arising from a hemizygous microdeletion at 7q11.23. The present study was focused on a comparative investigation of genomic integrity in WBS patients by use of cytogenetic methods and the alkaline comet assay. Lymphocytes of whole peripheral blood were cultured and metaphases were examined for frequency and spectrum of chromosome aberrations. A WBS-related microdeletion was detected by means of the FISH (fluorescence in situ hybridization) technique. The blood samples from patients who were carriers of this microdeletion, were tested in the comet assay. For this purpose, freshly collected lymphocytes were exposed to hydrogen peroxide (100μM, 1min, 4°C). The frequencies of endogenous and exogenous DNA damage, and the kinetics and efficiency of DNA repair were measured during three subsequent hours of incubation. Comparison of the two data sets in this group of patients demonstrated a slightly elevated average frequency of chromosome aberrations, significantly increased levels of endogenous and H(2)O(2)-induced DNA damage, and somewhat impaired DNA repair. The relationship between an abnormal DNA-damage response and the 7q11.23 hemizygous microdeletion was confirmed experimentally when comparing the comet assay data in FISH-positive and FISH-negative lymphocytes from WBS-suspected patients. Briefly, our results indicate the impact of chromosomal instability within this region on susceptibility towards DNA damage, which may contribute to pathogenesis of this disease. It was shown also that the comet assay, as well as an experimental design proposed here, seem to be useful tools for estimating genome integrity in WBS patients.

To determine conversion factors for the new International Commission on Radiological Protection (ICRP) publication 103 recommendations for adult and pediatric patients and to compare the effective doses derived from Monte Carlo calculations with those derived from dose-length product (DLP) for different body regions and computed tomographic (CT) scanning protocols.Effective dose values for the Oak Ridge National Laboratory phantom series, including phantoms for newborns; 1-, 5-, and 10-year-old children; and adults were determined by using Monte Carlo methods for a 64-section multidetector CT scanner. For each phantom, five anatomic regions (head, neck, chest, abdomen, and pelvis) were considered. Monte Carlo simulations were performed for spiral scanning protocols with different voltages. Effective dose was computed by using ICRP publication 60 and publication 103 recommendations. The calculated effective doses were compared with those derived from the DLP by using previously published conversion factors.In general, conversion factors determined on the basis of Monte Carlo calculations led to lower values for adults with both ICRP publications. Values up to 33% and 32% lower than previously published data were found for ICRP publication 60 and ICRP publication 103, respectively. For pediatric individuals, effective doses based on the Monte Carlo calculations were higher than those obtained from DLP and previously published conversion factors (eg, for chest CT scanning in 5-year-old children, an increase of about 76% would be expected). For children, a variation in conversion factors of up to 15% was observed when the tube voltage was varied. For adult individuals, no dependence on voltage was observed.Conversion factors from DLP to effective dose should be specified separately for both sexes and should reflect the new ICRP recommendations. For pediatric patients, new conversion factors specific for the spectrum used should be established.

One of the most important steps in achieving aesthetical and functional result in mandible reconstruction with fibula free flap is positioning of the bony part relative to the basis of the native mandible. Positioning fibular bony part continuing inferior mandible border may produce a high discrepancy between the residual mandible and fibula bone that results in difficulty in denture rehabilitation. Positioning fibular bony part continuing alveolar mandible border creates asymmetric jowls and changed shape of jawline in border-line zone. The double-barrel technique solves this problem, but needs a meticulous preoperative planning and precision free fibula flap modelling.При устранении дефектов нижней челюсти (НЧ) с помощью реваскуляризуемого малоберцового лоскута одним из ключевых моментов, влияющих на достижение функционального и эстетического результатов, является позиционирование малоберцовой кости (МБК) относительно интактной части нативной НЧ. Если малоберцовая кость располагается по нижнему краю (основанию) нативной НЧ, то высота несоответствия и мальпозиции относительно проекции альвеолярной части затрудняет процесс реабилитации функции зубочелюстной системы посредством изготовления ортопедической конструкции с опорой на дентальные имплантаты. Однако если малоберцовый трансплантат располагается по верхнему краю, в проекции альвеолярной части, в дальнейшем возникают эстетические проблемы в виде несимметричных провисающих 'брылей' или выраженных заломов мягких тканей на границе малоберцовая кость - нативная НЧ. Методика 'double barrel' (сдвоенной малоберцовой кости по типу 'двуствольного ружья с вертикальным расположением стволов - двустволки') позволяет решить данную проблему, но эта техника требует тщательного предоперационного планирования и точного интраоперационного моделирования.

The combining site specificities of IgE antibodies that react with the oral antibacterial agent trimethoprim and found in the sera of two subjects who experienced anaphylaxis after taking the drug, were investigated. Hapten inhibition studies with some close analogues of trimethoprim and a range of other structurally related compounds showed that the allergenic determinant complementary to the IgE antibodies in the serum of one of the subjects was the 3,4-dimethoxybenzyl group. The complementary allergenic structure recognized by the IgE antibodies in the serum from the second subject comprised both the trimethoxybenzyl and diaminopyrimidine rings of trimethoprim. Thus, as with thiopentone, but unlike the neuromuscular blocking drugs, the trimethoprim molecule has more than one determinant each with the capacity to provoke IgE formation, interact with the antibody combining site and provoke drug-induced allergic reactions. The general approach set out here employing carefully selected structural analogues in hapten inhibition studies should be invaluable for confirming specificity and identifying allergenic determinants in IgE antibody-mediated allergic drug reactions.

Sturge-Weber syndrome, also known as encephalotrigeminal angiomatosis or meningofacial angiomatosis, is characterised in its classical form by a congenital, usually unilateral, ‘portwine stain’ (capillary naevus) on the face, convulsions, typical intracranial calcification and some degree of mental retardation and hemiparesis. The clinical correlation of intractable seizures with the presence of bilateral intracranial disease has management and prognostic implications, thus making the presence of bilateral disease an important factor to all those involved in the management of the child with Sturge-Weber syndrome.

In a former publication the authors showed that low amounts of amorphous content (LOQ of 0.5%) in a hydrophobic model API (Ciclesonide) can be measured with an individually adjusted one-step dynamic organic vapor sorption (DVS). In this investigation the applicability is tested on various APIs which differ in lipophilicity (poor water solubility) and hygroscopicity (absorption of water). The vapor sorption method proved to be applicable in almost all cases. Moisture sorption isotherms were determined for all five investigated crystalline and amorphous APIs. However, it was necessary to select the parameters individually for each API. The used solvents (water, methanol, isopropanol and methylene chloride) and the humidity-levels (0.05 p/p0 until 0.5 p/p0) were chosen carefully because otherwise the amorphous amounts switch to their crystalline counterparts and are not detectable. The production of fully amorphous samples (absence of crystalline material measured by DSC, mDSC and XRPD) was optimized over several trials. As successfully methods proved ball-milling, freeze-drying, spray-drying and/or quench cooling. In the next step these fully amorphous amounts were blended with crystalline starting material to calibration curves (Turbula blender, influence of electrostatic charge to homogeneity) for the calculation of amorphous content. In summary, the following presented methods were used to determine and quantify low amorphous amounts (between 1.5% and 17.0%) in jet-milled powders (grinding pressure of 8bar, 1-3 grinding cycles), respectively.

Integration of pre- or peri-operative images may improve image guidance in minimally invasive interventions. In abdominal catheterization procedures such as transcatheter arterial chemoembolization, 3D pre-/peri-operative images contain relevant information, such as complete 3D vasculature, that is not directly available from 2D imaging. Accurate knowledge of the catheter tip position in 3D is currently not available, and after registration of 3D information to 2D images (angiographies), the registration is invalidated by breathing motion and thus requires continuous updates. We propose a hidden Markov model based method to track the 3D catheter position, using 2D fluoroscopic image sequences and a 3D vessel tree obtained from 3D Rotational Angiography. Such a tracking facilitates display of the catheter in the 3D anatomy, and it enables to use the 3D vessels as a roadmap in 2D imaging. The tracking is initialized with the first 2D image of the sequence. For the subsequent images, based on a state transition probability distribution and the registration observations, the catheter tip position is tracked in the 3D vessel tree using registrations to the 2D fluoroscopic images. The method is evaluated on simulated data and two clinical sequences. In the simulations, we obtain a median tip position accuracies up to 2.9 mm. On clinical sequence, the distance between the catheter and the projected vessels after registration is below 1.9 mm.

2-Chloro-2'-deoxyadenosine (CdA) is a nucleoside analogue active in B-cell chronic lymphocytic leukemia (B-CLL). Although the mechanism of action of CdA has been extensively investigated in leukemic cells, the possibility that this nucleoside analogue interacts with the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway has never been explored. In this study, we show that CdA, at concentrations close to the IC50, activated the ERK pathway in the B-cell line EHEB. Because activation of this pathway is assumed to exert anti-apoptotic effect, we combined CdA with inhibitors of the ERK pathway. The latter were found to enhance CdA-induced apoptosis. These results suggest that the efficacy of CdA could be strengthened by combination with inhibitors of the ERK pathway.

Monoclonal antibodies are widely used at GlaxoSmithKline Biologicals (GSK Bio) for the quantification and characterization of antigens and for the release of vaccine lots. In 1998, GSK Bio decided to change the production of monoclonal antibodies (MAbs) designed for immunological tools from in vivo to in vitro technology. In 2004, all MAbs used at GSK Bio were produced in vitro. These MAbs cover more than 100 different targets with a variety of 1500 hybridomas, and approximately 60 to 90 MAbs are produced every year. This article describes the development process, including a description of the different systems tested based on double membrane or hollow fiber technology. The productivity, assets, and drawbacks of the different technologies are presented, and evaluation strategies for the choice of in vitro systems are discussed. Binding kinetics displayed by MAbs produced in vitro and in vivo were found to be similar, and MAbs produced in vitro are suitable tools for various immunological applications.

Moyamoya is a rare cerebrovascular disease characterised by progressive stenosis of the terminal portion of the internal carotid artery and its main branches. We report on the progression of the disease in a 3-year-old boy, confirmed to have idiopathic Moyamoya disease, over 6 ensuing years, using serial magnetic resonance imaging with MR angiography. Comparison is also made with conventional angiography at the last visit. Characteristic imaging appearances of asymmetric narrowing of the internal carotid arteries (especially the supra-clinoid portion) with numerous collateral vessels around the brainstem especially within the ambient and quadrigeminal plate cisterns are demonstrated with progression.