Face-off Droop: A Case Report of Pediatric Stroke.

Introduction: Cerebrovascular accidents rarely occur in children; the incidence of ischemic stroke in patients <16 years of age is between 0.6-7.9/100,000. However, they are the fourth most common cause of acute neurological deficits in the pediatric population, and possible cases should be evaluated with a high index of suspicion to ensure timely intervention. Case Report: We describe a previously healthy 17-year-old male who presented to the pediatric emergency department with a left facial droop and hemiparesis consistent with a stroke. The patient's age and lack of comorbidities made this an extremely uncommon presentation. Our patient's neurologic symptoms were believed to have been caused by a recent traumatic clavicular injury sustained two weeks prior, which subsequently led to vascular insult. Conclusion: Cerebrovascular accidents are an important cause of morbidity and mortality in pediatric patients. Cerebrovascular accidents in children are most often secondary to congenital causes; however, care should be taken to assess for acquired causes, such as trauma to major blood vessels. While rarely implicated in traumatic injuries, arterial structures posterior to the medial clavicle can result in severe complications.

Barley Seeds miRNome Stability during Long-Term Storage and Aging.

Seed aging is a complex biological process that has been attracting scientists' attention for many years. High-throughput small RNA sequencing was applied to examine microRNAs contribution in barley seeds senescence. Unique samples of seeds that, despite having the same genetic makeup, differed in viability after over 45 years of storage in a dry state were investigated. In total, 61 known and 81 novel miRNA were identified in dry seeds. The highest level of expression was found in four conserved miRNA families, i.e., miR159, miR156, miR166, and miR168. However, the most astonishing result was the lack of significant differences in the level of almost all miRNAs in seed samples with significantly different viability. This result reveals that miRNAs in dry seeds are extremely stable. This is also the first identified RNA fraction that is not deteriorating along with the loss of seed viability. Moreover, the novel miRNA hvu-new41, with higher expression in seeds with the lowest viability as detected by RT-qPCR, has the potential to become an indicator of the decreasing viability of seeds during storage in a dry state.

Genes ScBx1 and ScIgl-Competitors or Cooperators?

Two genes, Bx1 and Igl, both encoding indole-3-glycerol phosphate lyase (IGL), are believed to control the conversion of indole-3-glycerol phosphate (IGP) to indole. The first of these has generally been supposed to be regulated developmentally, being expressed at early stages of plant development with the indole being used in the benzoxazinoid (BX) biosynthesis pathway. In contrast, it has been proposed that the second one is regulated by stresses and that the associated free indole is secreted as a volatile. However, our previous results contradicted this. In the present study, we show that the ScIgl gene takes over the role of ScBx1 at later developmental stages, between the 42nd and 70th days after germination. In the majority of plants with silenced ScBx1 expression, ScIgl was either expressed at a significantly higher level than ScBx1 or it was the only gene with detectable expression. Therefore, we postulate that the synthesis of indole used in BX biosynthesis in rye is controlled by both ScBx1 and ScIgl, which are both regulated developmentally and by stresses. In silico and in vivo analyses of the promoter sequences further confirmed our hypothesis that the roles and modes of regulation of the ScBx1 and ScIgl genes are similar.

An open-label phase 1 clinical trial of the anti-α4ß7 monoclonal antibody vedolizumab in HIV-infected individuals.

Despite the substantial clinical benefits of antiretroviral therapy (ART), complete eradication of HIV has not been possible. The gastrointestinal tract and associated lymphoid tissues may play an important role in the pathogenesis of HIV infection. The integrin α4ß7 facilitates homing of T lymphocytes to the gut by binding to the mucosal addressin cell adhesion molecule-1 (MAdCAM-1) expressed on venules in gut-associated lymphoid tissue. CD4+ T cells with increased expression of α4ß7 are susceptible to HIV infection and may be key players in subsequent virus dissemination. Data from nonhuman primate models infected with simian immunodeficiency virus (SIV) have suggested that blockade of the α4ß7/MAdCAM-1 interaction may be effective at preventing SIV infection and may have beneficial effects in animals with established viral infection. To explore whether these findings could be reproduced in HIV-infected individuals after interruption of ART, we conducted an open-label phase 1 clinical trial of vedolizumab, a monoclonal antibody against α4ß7 integrin. Vedolizumab infusions in 20 HIV-infected individuals were well tolerated with no serious adverse events related to the study drug. After interruption of ART, the median time to meeting protocol criteria to restart therapy was 13 weeks. The median duration of plasma viremia of <400 copies/ml was 5.4 weeks. Only a single subject in the trial experienced prolonged suppression of plasma viremia after interruption of ART. These results suggest that blockade of α4ß7 may not be an effective strategy for inducing virological remission in HIV-infected individuals after ART interruption.

The miRNAome dynamics during developmental and metabolic reprogramming of tomato root infected with potato cyst nematode.

Cyst-forming plant-parasitic nematodes are pests threatening many crops. By means of their secretions cyst nematodes induce the developmental and metabolic reprogramming of host cells that lead to the formation of a syncytium, which is the sole food source for growing nematodes. The in depth micro RNA (miRNA) dynamics in the syncytia induced by Globodera rostochiensis in tomato roots was studied. The miRNAomes were obtained from syncytia covering the early and intermediate developmental stages, and were the subject of differential expression analysis. The expression of 1235 miRNAs was monitored. The fold change (log2FC) ranged from -7.36 to 8.38, indicating that this transcriptome fraction was very variable. Moreover, we showed that the DE (differentially expressed) miRNAs do not fully overlap between the selected time points, suggesting infection stage specific regulation by miRNA. The correctness of RNA-seq expression profiling was confirmed by qRT-PCR (quantitative Real Time Polymerase Chain Reaction) for seven miRNA species. Down- and up-regulated miRNA species, including their isomiRs, were further used to identify their potential targets. Among them there are a large number of transcription factors linked to different aspects of plant development belonging to gene families, such as APETALA2 (AP2), SQUAMOSA (MADS-box), MYB, GRAS, and AUXIN RESPONSE FACTOR (ARF). The substantial portion of potential target genes belong to the NB-LRR and RLK (RECEPTOR-LIKE KINASE) families, indicating the involvement of miRNA mediated regulation in defense responses. We also collected the evidence for target cleavage in the case of 29 miRNAs using one of three alternative methods: 5' RACE (5' Rapid Amplification of cDNA Ends), a search of tasiRNA within our datasets, and the meta-analysis of tomato degradomes in the GEO (Gene Expression Omnibus) database. Eight target transcripts showed a negative correlation with their respective miRNAs at two or three time points. These results indicate a large regulatory potential for miRNAs in tuning the development and defense responses.

FAST MRI breast screening revisited.

INTRODUCTION: Screening for breast cancer in high-risk women takes about 40 minutes to acquire an MRI scan and is time-intensive to report. There is recent interest in the performance of an abbreviated MRI protocol (FAST) in the screening setting. FAST scans have a reported negative predictive value of 99.8%. This study evaluates the false positive rates (FPR) and recall rates for FAST scans as compared to full diagnostic studies (FD). METHODS: A database of all screening breast MRI scans performed at our institution between 30 June 2013 and 1 July 2014 (n = 591) was created by one of the researchers, who did not subsequently analyse the MRI scans. The T1W and first post-contrast and subtracted images from each of these scans (FAST protocol) were assessed by experienced breast MRI radiologists, blinded to the final diagnosis. The findings were then compared with the FD result. RESULTS: The recall rates were 6.6% for FAST scans and 5.8% for FD scans. FPR rates were 4.7% and 3.9% respectively. There is no statistically significant difference in the recall rates or FPR of FAST scans in comparison with full diagnostic studies. CONCLUSIONS: Given the absence of statistically significant difference in the FPR and recall rates in comparison with FD, FAST scans can replace FD for screening of breast cancer.

Changes in Serum Amino Acids in Migraine Patients without and with Aura and their Possible Usefulness in the Study of Migraine Pathogenesis.

BACKGROUND: Results of several studies suggest that serum amino acids monitoring in migraine might be useful as an objective measurement of the disease status. OBJECTIVE: The aim of the present work was to analyze the profile of aliphatic and aromatic amino acids in blood serum of migraine patients without and with aura between attacks. METHODS: A total number of 37 migraine patients (26 with migraine without aura and 11 with migraine with aura), mean age 39±12 years, and 40 age-matched healthy subjects as the control group, mean age 38±14 years, were included into the study. The levels of glutamic acid, glutamine, histidine (His), valine (Val), isoleucine, leucine (Leu), phenyloalanine, lysine were evaluated. RESULTS: The level of His was significantly higher in both groups of migraine patients (without and with aura) compared to the control group (F(2,74)=10.17, p=0.00). The levels of Val and Leu were significantly different in migraine without but not with aura, when compared with the control group (F(2,74)=4.70, p=0.01 and F(2,74)=4.39, p=0.02, respectively). CONCLUSION: We found higher level of His in migraine patients without and with aura and lower level of Val and Leu in patients with migraine without aura.

[Hyperventilation echocardiography in vasospastic angina pectoris diagnosing]. / Hyperventilacní echokardiografie v diagnostice vazospastické anginy pectoris.

Hyperventilation echocardiography is an established diagnostic test in patients with suspected variant angina pectoris. It has got sufficient sensitivity (60-80%) and specificity (85-100%). Positive hyperventilation test is rarely found, which relates to low prevalence of variant angina. The diagnostic yield of the test depends on the population selected for testing: positive result can be expected in patients with a history of typical burning chest pain, ST segment elevation/depression and/or inversions of U wave during the chest pain episode, arrhythmias related to the chest pain, coronary artery stenosis less than 50% of artery diameter, multi-vessel disease, high activity of illness at the time of hyperventilation test. We present a case of 37 years old man with typical angina pectoris at rest and non-Q myocardial infarction, in whom the coronary angiography was negative. Variant angina pectoris was diagnosed by hyperventilation echocardiography. The ECG tracings showing typical ischemic patterns during the hyperventilation test are included.

[Infective endocarditis--crucial early diagnosis]. / Infekcní endokarditida--klícová vcasná diagnóza.

A 64-year old female patient with polymorbidity, including diabetes, was admitted to our clinic with a suspicion of silent myocardial infarction. However, no acute coronary lesion was subsequently confirmed. Transthoracal echocardiography performed just after the admission suggested infective endocarditis. The diagnosis was difficult due to a lack of collaboration by the patient who was disoriented and confused. Because of the suspicion ofendocarditis, a therapy with antibiotics was started immediately and the diagnosis was confirmed by oesophageal echocardiography and heart MR performed after the patient was stabilized. An early diagnosis and an intensive antibiotic treatment are crucial for the management of the frequently fatal infective endocarditis.

Mutations in GBA are associated with familial Parkinson disease susceptibility and age at onset.

OBJECTIVE: To characterize sequence variation within the glucocerebrosidase (GBA) gene in a select subset of our sample of patients with familial Parkinson disease (PD) and then to test in our full sample whether these sequence variants increased the risk for PD and were associated with an earlier onset of disease. METHODS: We performed a comprehensive study of all GBA exons in one patient with PD from each of 96 PD families, selected based on the family-specific lod scores at the GBA locus. Identified GBA variants were subsequently screened in all 1325 PD cases from 566 multiplex PD families and in 359 controls. RESULTS: Nine different GBA variants, five previously reported, were identified in 21 of the 96 PD cases sequenced. Screening for these variants in the full sample identified 161 variant carriers (12.2%) in 99 different PD families. An unbiased estimate of the frequency of the five previously reported GBA variants in the familial PD sample was 12.6% and in the control sample was 5.3% (odds ratio 2.6; 95% confidence interval 1.5-4.4). Presence of a GBA variant was associated with an earlier age at onset (p = 0.0001). On average, those patients carrying a GBA variant had onset with PD 6.04 years earlier than those without a GBA variant. CONCLUSIONS: This study suggests that GBA is a susceptibility gene for familial Parkinson disease (PD) and patients with GBA variants have an earlier age at onset than patients with PD without GBA variants.

"TOF2H": a precision toolbox for rapid, high density/high coverage hydrogen-deuterium exchange mass spectrometry via an LC-MALDI approach, covering the data pipeline from spectral acquisition to HDX rate analysis.

BACKGROUND: Protein-amide proton hydrogen-deuterium exchange (HDX) is used to investigate protein conformation, conformational changes and surface binding sites for other molecules. To our knowledge, software tools to automate data processing and analysis from sample fractionating (LC-MALDI) mass-spectrometry-based HDX workflows are not publicly available. RESULTS: An integrated data pipeline (Solvent Explorer/TOF2H) has been developed for the processing of LC-MALDI-derived HDX data. Based on an experiment-wide template, and taking an ab initio approach to chromatographic and spectral peak finding, initial data processing is based on accurate mass-matching to fully deisotoped peaklists accommodating, in MS/MS-confirmed peptide library searches, ambiguous mass-hits to non-target proteins. Isotope-shift re-interrogation of library search results allows quick assessment of the extent of deuteration from peaklist data alone. During raw spectrum editing, each spectral segment is validated in real time, consistent with the manageable spectral numbers resulting from LC-MALDI experiments. A semi-automated spectral-segment editor includes a semi-automated or automated assessment of the quality of all spectral segments as they are pooled across an XIC peak for summing, centroid mass determination, building of rates plots on-the-fly, and automated back exchange correction. The resulting deuterium uptake rates plots from various experiments can be averaged, subtracted, re-scaled, error-barred, and/or scatter-plotted from individual spectral segment centroids, compared to solvent exposure and hydrogen bonding predictions and receive a color suggestion for 3D visualization. This software lends itself to a "divorced" HDX approach in which MS/MS-confirmed peptide libraries are built via nano or standard ESI without source modification, and HDX is performed via LC-MALDI using a standard MALDI-TOF. The complete TOF2H package includes additional (eg LC analysis) modules. CONCLUSION: "TOF2H" provides a comprehensive HDX data analysis package that has accelerated the processing of LC-MALDI-based HDX data in the authors' lab from weeks to hours. It runs in a standard MS Windows (XP or Vista) environment, and can be downloaded http://tof2h.bio.uci.edu or obtained from the authors at no cost.

[Chronic benign exsudative pericarditis--a case of familiar incidence]. / Chronická exsudativní benigní perikarditida--popis familiárního výskytu.

Pericarditis is a common disease caused by a number of factors. Chronic pericarditis is defined as the presence of pericardial effusion for more than 3 or 6 months. The case study reports a case of familiar incidence of chronic exsudative pericarditis in a young woman, her sister and her mother, with an analysis of diagnostic and therapeutic options. According to available literature, this is the second case described of such form of familiar incidence.

LRRK2 mutation analysis in Parkinson disease families with evidence of linkage to PARK8.

BACKGROUND: Pathogenic mutations in the leucine-rich repeat kinase 2 gene (LRRK2) have been found to cause typical, later-onset Parkinson disease (PD). Although G2019S is the most common mutation, other mutations have also been reported. It is critical to catalog the types of mutations found in LRRK2 that can cause PD, so as to provide insight regarding disease susceptibility and potential novel treatments. METHODS: We performed a comprehensive study of all 51 exons of the LRRK2 gene in one PD patient from each of 88 multiplex PD families who had the highest family-specific multipoint lod score at the LRRK2 locus from a cohort of 430 PD families without the G2019S mutation. RESULTS: Five families (5.7%) harbored what seem to be novel, pathogenic mutations (L1795F, I1192V, E10K, E334K, Q1111H). Three of these apparent mutations were in known, functional domains of the LRRK2 protein, where other studies have also identified disease producing mutations. However, two of the novel variants were found in the N-terminal region of LRRK2, where no pathogenic substitutions have yet been reported. Similar to previous studies, all subjects with an LRRK2 mutation had classic symptoms of PD and typical, later age at onset. CONCLUSIONS: We have identified five novel variants in LRRK2, with two of these in the N-terminal region of LRRK2, where no pathogenic substitutions have been previously reported. If confirmed to be causative, these mutations would broaden the potential mechanisms whereby mutations in LRRK2 result in Parkinson disease.

[Long term results of cardiac resynchronisation therapy for patients with severe heart failure]. / Dlouhodobé výsledky srdecní resynchronizacní lécby u pacientu s tezkým srdecním selhaním.

INTRODUCTION: Cardiac resynchronisation therapy (CRT) has been shown to be a highly effective treatment strategy for patients with severe chronic heart failure (ChHF). OBJECTIVE OF STUDY: To determine the clinical response of patients to CRT, to measure morbidity and mortality for this population of patients, to determine causes and predictors of death. To test whether the parameters of tissue Doppler echocardiography are able to predict response to CRT. PATIENTS AND METHODS: Before and after implantation of the CRT and 12 months later, echocardiograph tests were carried out and relevant clinical data was recorded during the monitoring of patients. RESULTS: 102 patients (71 men, 31 women) with an average age of 71 +/- 9 years took part in the study. 68% patients had cardiac ischemia, 29% had idiopathic dilated cardiomyopathy. 75% patients were in functional class NYHA III, 25% NYHA IV. After a monitoring period of 711 +/- 329 days, 26 patients had died and 35 patients had been hospitalised. 34% of all hospitalisations were for acute exacerbation of ChHF. Patients with initial functional classification NYHA IV had a higher mortality rate in years one and two than patients in class NYHA III. The proportion of clinical responders was 64% after 12 months of CRT. In 58% of patients, a year of CRT produced a relative increase in the ejection fraction of the left ventricle (EF LV) of > or = 30%. 1/3 of patients had EF LV ? 45% with minimal symptoms of ChHF. The following were found to predict reverse remodelling of the left ventricle: less advanced state of the basic illness (EFLV > 23%, left ventricular end-diastolic diameter < 65 mm, left ventricular end-diastolic volume < 160 ml and left ventricular end-systolic volume < 120 ml) and interventricular mechanical delay > 45 ms. CONCLUSION: CRT is a safe method with a high success rate. There continues to be a problem with identifying responders. Symptoms of less advanced heart disease and interventricular delay were identified as sensitive predictors of the response to treatment.

Acute ischemia as a complication by transoesophageal echocardiography.

Transoesophageal echocardiography (TEE) is a safe procedure with a low complication rate. We present a patient with severe coronary artery stenosis or disease who developed acute coronary ischemia of the anterior wall as a complication of TEE procedure. The possible mechanism can be stress during the procedure.

Differential detergent fractionation for non-electrophoretic eukaryote cell proteomics.

Differential detergent fractionation (DDF), which relies on detergents to sequentially extract proteins from eukaryotic cells, has been used to increase proteome coverage of 2D-PAGE. Here, we used DDF extraction in conjunction with the nonelectrophoretic proteomics method of liquid chromatography and electrospray ionization tandem mass spectrometry. We demonstrate that DDF can be used with 2D-LC ESI MS2 for comprehensive cellular proteomics, including a large proportion of membrane proteins. Compared to some published methods designed to isolate membrane proteins specifically, DDF extraction yields comprehensive proteomes which include twice as many membrane proteins. Two-thirds of these membrane proteins have more than one trans-membrane domain. Since DDF separates proteins based upon their physicochemistry and subcellular localization, this method also provides data useful for functional genome annotation. As more genome sequences are completed, methods which can aid in functional annotation will become increasingly important.

[Acute myocardial infarct as a result of external compression caused by an expanding pulmonary adenocarcinoma]. / Akutní infarkt myokardu jako dusledek zevní komprese prorustajícího adenokarcinomu plic.

The case report describes a rare cause of acute myocardial infarction. 68-years old man with the cancer triplicity (follicular and papillar carcinoma of the thyroid gland, Grawitz tumor, bronchogenic carcinoma with generalization) was admitted to the intensive care unit for the sudden onset of chest pain, positivity of cardioselective enzymes and signs of cardiogenic shock. Echocardiographic examination proved diaphragmatic akinesis with low left ventricular ejection fraction about 30%. The acute coronary angiogram revealed unimportant atherosclerotic narrowing of the left coronary artery and a long significant stenosis in the proximal portion of the right coronary artery untypical for atherosclerotic lesion suspicious of extramural compression. A pathological vascularization to the extramyocardial region was documented during right coronary angiogram. The patient was treated conservatively and finally transferred to the pulmonary disease department, where he died of progression of the cancer disease two months later. Postmortem examination found spreading of the bronchogenic adenocarcinoma to the pericardium along the course of the right coronary artery, but neither direct infiltration of the vessel wall by the tumor nor atherosclerotic disease of the proximal portion of the right coronary artery were proved. These findings together with the coronary angiogram demonstrate a rare cause of the myocardial infarction due to the extramural compression by the malignant tumor.

Successful Day 5 embryo transfer and pregnancies resulting after transport of embryos by air for biopsy and genetic analysis.

PURPOSE: Case studies of four in vitro fertilization (IVF) cycles where embryo transport by commercial airline followed by biopsy and genetic analysis with subsequent culture to Day 5 and resulting ongoing pregnancy. METHOD: Retrospective clinical case study of 4 patients requiring preimplantation genetic diagnosis (PGD) testing. Normally fertilized embryos were transported in a battery-powered portable incubator by commercial airline following evaluation for fertilization under controlled conditions from the Center for Assisted Reproduction, Bedford, Texas to the Reproductive Genetic Institute, Chicago, Illinois. Following Day 3 embryo biopsy and genetic analysis, embryos were transported back to the Center for Assisted Reproduction for Day 5 embryo transfer. RESULTS: Ongoing clinical pregnancy resulted for all patients receiving embryo transfer. CONCLUSION: These results demonstrate the feasibility of embryo transport by air for centers that do not have the in-house capabilities to perform genetic analysis. With successful pregnancies obtained through extended culture to Day 5, embryos requiring genetic analysis can be successfully transported by air, tested, and returned to the initial facility for embryo transfer without time restriction.