Insulin sensitizer prevents and ameliorates experimental type 1 diabetes.
Insulin-dependent type-1 diabetes (T1D) is driven by autoimmune β-cell failure, whereas systemic resistance to insulin is considered the hallmark of insulin-independent type-2 diabetes (T2D). In contrast to this canonical dichotomy, insulin resistance appears to precede the overt diabetic stage of T1D and predict its progression, implying that insulin sensitizers may change the course of T1D. However, previous attempts to ameliorate T1D in animal models or patients by insulin sensitizers have largely failed. Sensitization to insulin by MEthyl-substituted long-chain DICArboxylic acid (MEDICA) analogs in T2D animal models surpasses that of current insulin sensitizers, thus prompting our interest in probing MEDICA in the T1D context. MEDICA efficacy in modulating the course of T1D was verified in streptozotocin (STZ) diabetic rats and autoimmune nonobese diabetic (NOD) mice. MEDICA treatment normalizes overt diabetes in STZ diabetic rats when added on to subtherapeutic insulin, and prevents/delays autoimmune T1D in NOD mice. MEDICA treatment does not improve β-cell insulin content or insulitis score, but its efficacy is accounted for by pronounced total body sensitization to insulin. In conclusion, potent insulin sensitizers may counteract genetic predisposition to autoimmune T1D and amplify subtherapeutic insulin into an effective therapeutic measure for the treatment of overt T1D.


Verbal Fluency Performance in Patients with Non-demented Parkinson's Disease.
While Parkinson's disease (PD) has traditionally been defined by motor symptoms, many researches have indicated that mild cognitive impairment is common in non-demented PD patients. The purpose of this study was to compare verbal fluency performance in non-demented Parkinson's disease patients with healthy controls. In this cross-sectional study thirty non-demented Parkinson's disease patients and 30 healthy controls, matched by age, gender and education, were compared on verbal fluency performance. Verbal fluency was studied with a Phonemic Fluency task using the letters F, A, and S, a semantic fluency task using the categories animals and fruits. The independent t-test was used for data analysis. Overall, participants generated more words in the semantic fluency task than in the phonemic fluency task. Results revealed significant differences between patients and controls in semantic fluency task (p<.05). In addition, PD patients showed a significant reduction of correctly generated words in letter fluency task. The total number of words produced was also significantly lower in the PD group (p<.05). Verbal fluency disruption is implied in non-demented PD patients in association with incipient cognitive impairment.


In vitro antiprotozoal activity of extracts of five Turkish Lamiaceae species.
The in vitro antiprotozoal activities of crude methanolic extracts from the aerial parts of five Lamiaceae plants (Salvia tomentosa, S. sclarea, S. dichroantha, Nepeta nuda subsp. nuda and Marrubium astracanicum subsp. macrodon) were evaluated against four parasitic protozoa, i.e. Trypanosoma brucei rhodesiense, T. cruzi, Leishmania donovani and Plasmodium falciparum. The cytotoxic potentials of the extracts on L6 cells were also evaluated. Melarsoprol, benznidazole, miltefosine, chloroquine and podophyllotoxin were used as reference drugs. All crude MeOH extracts showed antiprotozoal potential against at least three parasites, so they were dispersed in water and partitioned against n-hexane and chloroform to yield three subextracts that were screened in the same test systems. The n-hexane extract of N. nuda was the most active against T. brucei rhodesiense while the CHCl3 extracts of S. tomentosa and S. dichroantha showed significant activity against L. donovani. All organic extracts displayed in vitro antimalarial and moderate trypanocidal activities against T. cruzi with the n-hexane extract of S. sclarea being the most active against the latter. The extracts displayed low or no cytotoxicity towards mammalian L6 cells.


Active Ebola Virus Replication and Heterogeneous Evolutionary Rates in EVD Survivors.
Following cessation of continuous Ebola virus (EBOV) transmission within Western Africa, sporadic EBOV disease (EVD) cases continued to re-emerge beyond the viral incubation period. Epidemiological and genomic evidence strongly suggests that this represented transmission from EVD survivors. To investigate whether persistent infections are characterized by ongoing viral replication, we sequenced EBOV from the semen of nine EVD survivors and a subset of corresponding acute specimens. EBOV evolutionary rates during persistence were either similar to or reduced relative to acute infection rates. Active EBOV replication/transcription continued during convalescence, but decreased over time, consistent with viral persistence rather than viral latency. Patterns of genetic divergence suggest a moderate relaxation of selective constraints within the sGP carboxy-terminal tail during persistent infections, but do not support widespread diversifying selection. Altogether, our data illustrate that EBOV persistence in semen, urine, and aqueous humor is not a quiescent or latent infection.


TiO2 Nanoparticle-Induced Oxidation of the Plasma Membrane: Importance of the Protein Corona.
Titanium dioxide (TiO2) nanoparticles, used as pigments and photocatalysts, are widely present in modern society. Inhalation or ingestion of these nanoparticles can lead to cellular-level interactions. We examined the very first step in this cellular interaction, the effect of TiO2 nanoparticles on the lipids of the plasma membrane. Within 12 h of TiO2 nanoparticle exposure, the lipids of the plasma membrane were oxidized, determined with a malondialdehyde assay. Lipid peroxidation was inhibited by surface passivation of the TiO2 nanoparticles, incubation with an antioxidant (Trolox), and the presence of serum proteins in solution. Subsequent experiments determined that serum proteins adsorbed on the surface of the TiO2 nanoparticles, forming a protein corona, inhibit lipid peroxidation. Super-resolution fluorescence microscopy showed that these serum proteins were clustered on the nanoparticle surface. These protein clusters slow lipid peroxidation, but by 24 h, the level of lipid peroxidation is similar, independent of the protein corona or free serum proteins. Additionally, over 24 h, this corona of proteins was displaced from the nanoparticle surface by free proteins in solution. Overall, these experiments provide the first mechanistic investigation of plasma membrane oxidation by TiO2 nanoparticles, in the absence of UV light and as a function of the protein corona, approximating a physiological environment.


Human activity as a manifold valued random process.
Most of previous shape based human activity models were built with either a linear assumption or an extrinsic interpretation of the nonlinear geometry of the shape space, both of which proved to be problematic on account of the nonlinear intrinsic geometry of the associated shape spaces. In this paper we propose an intrinsic stochastic modeling of human activity on a shape manifold. More importantly, within an elegant and theoretically sound framework, our work effectively bridges the nonlinear modeling of human activity on a nonlinear space, with the classic stochastic modeling in a Euclidean space, and thereby provides a foundation for a more effective and accurate analysis of the nonlinear feature space of activity models. From a video sequence, human activity is extracted as a sequence of shapes. Such a sequence is considered as one realization of a random process on a shape manifold. Different activities are then modeled as manifold valued random processes with different distributions. To address the problem of stochastic modeling on a manifold, we first construct a nonlinear invertible map of a manifold valued process to a Euclidean process. The resulting process is then modeled as a global or piecewise Brownian motion. The mapping from a manifold to a Euclidean space is known as a stochastic development. The advantage of such a technique is that it yields a one-one correspondence, and the resulting Euclidean process intrinsically captures the curvature on the original manifold. The proposed algorithm is validated on two activity databases [15], [5] and compared with the related works on each of these. The substantiating results demonstrate the viability and high accuracy of our modeling technique in characterizing and classifying different activities.


Altered lateralisation of emotional prosody processing in schizophrenia.
Alterations of cerebral lateralisation in schizophrenia have been reported consistently, and a reduced left-lateralisation has been suggested for language functions. Speech contains non-verbal information, e.g. prosody, and on a behavioural level, the extraction of emotional information from prosody is often impaired in schizophrenia. A previous functional magnetic resonance imaging study suggests increased left-lateralisation in schizophrenia during prosody processing, but did not disentangle effects of speech processing as such and emotional prosody processing. Here, we used meaningless syllables spoken with neutral, angry or fearful speech melody and measured blood oxygen level-dependent (BOLD) responses in 15 in-patients with schizophrenia and 15 healthy control participants matched for age and gender. Lateralisation indices were calculated for responses to emotional versus neutral prosody, and for all types of prosody versus baseline. Compared to control participants, patients with schizophrenia showed an increased right-lateralisation of emotional and non-emotional prosody processing in the temporal and parietal cortex. This right-lateralisation was increased in patients with reduced right-handedness and decreased in patients with stronger negative symptoms, particularly affective blunting, and with longer hospitalisation. Although patients with schizophrenia performed worse in emotion identification, this deficit was not related to lateralisation indices. Enhanced right-lateralisation to prosody resembles previous findings on laterality changes in speech processing and might suggest a common underlying cause in the organization of language functions.


A conceptual model of quality of life in schizophrenia: description and preliminary clinical validation.
The utility of quality of life (QOL) as an evaluative tool in clinical psychiatric research and drug trials could be enhanced by developing appropriate conceptual models of QOL, specific for psychiatric disorders. In our proposed model, QOL of individuals maintained on antipsychotic drug therapy for schizophrenia, is viewed as the subject's perception of the outcome of an interaction between severity of psychotic symptoms, side-effects including subjective responses to antipsychotic drugs, and the level of psychosocial performance. In order to test the validity of the model in clinical setting, we selected a sample of 62 schizophrenic patients clinically stabilized on antipsychotic drug therapy, and measured their subjective QOL and other potentially relevant clinical and psychosocial factors. Standardized scales including the positive and negative syndromes scale (PANSS), abnormal involuntary movements scale (AIMS), Hillside Akathisia scale (HAI), and the social performance schedule (SPS) were used for this purpose. Results of a multiple regression analysis using subjective quality of life as the outcome variable, indicated that severity of schizophrenic symptoms (partial R2 = 0.32, p < 0.0001) and subjective distress caused by akathisia (partial R2 = 0.11, p < 0.01) and neuroleptic dysphoria (partial R2 = 0.06, p < 0.05), accounted for nearly half of the variance, while the contribution from the psychosocial indicators was negligible. These results broadly endorse key aspects of the proposed model, and suggest further studies in this direction. These results experiences during antipsychotic therapy can enhance patients' QOL. This conceptual model has been developed with particular focus on the impact of antipsychotic medications on the QOL of persons with schizophrenia. As such, it is more applicable to clinical trials of new antipsychotic medications but may not be broad enough to be applicable for other social or vocational interventions.


Asperger's syndrome: to be or not to be?
It has been questioned whether Asperger's syndrome (AS) is in fact a specific (high functioning) subgroup of autism, rather than a distinct entity. Thirteen AS patients were compared with 13 autistic patients and 13 developmentally disordered controls. While there was symptom overlap between AS and autism, patients could be separated into one or other group. However, current criteria are based on symptoms, and it is argued that studies of genetics and treatment response are needed to elucidate the relationship between these developmental disorders.


Detection of Salmonella in meat: comparative and interlaboratory validation of a noncomplex and cost-effective pre-PCR protocol.
Cost-effective and rapid monitoring of Salmonella in the meat production chain can contribute to food safety. The objective of this study was to validate an easy-to-use pre-PCR sample preparation method based on a simple boiling protocol for screening of Salmonella in meat and carcass swab samples using a real-time PCR method. The protocol included incubation in buffered peptone water, centrifugation of an aliquot, and a boiling procedure. The validation study included comparative and interlaboratory trials recommended by the Nordic Organization for Validation of Alternative Microbiological Methods (NordVal). The comparative trial was performed against a reference method (NMKL 187, 2007) and a PCR method previously approved by NordVal with a semiautomated magnetic bead-based DNA extraction step using 122 artificially contaminated samples. The LOD was found to be 3.0, 3.2, and 3.4 CFU/sample for the boiling, magnetic bead-based, and NMKL 187 methods, respectively. When comparing the boiling method with the magnetic beads, the relative accuracy (AC), relative sensitivity (SE), and relative specificity (SP) were 98, 102, and 98%, respectively (Cohen's kappa index 0.95). When comparing results obtained by the boiling to the culture-based method, the AC, SE, and SP were found to be 98, 102, and 98%, respectively (kappa index 0.93). In the interlaboratory trial including valid results from 11 laboratories, apart from two false-positive samples by the boiling method combined with PCR, no deviating results were obtained (SP, SE, and AC were 100, 95, and 97%, respectively). This test is under implementation by the Danish meat industry, and can be useful for screening of large number of samples in the meat production, especially for fast release of minced meat with a short shelf life.


Late sinus and atrial tachycardia after pediatric heart transplantation might predict poor outcome.
Our objective was to examine clinical/electrocardiogram (ECG) predictors and outcomes of arrhythmias beyond 1 year after pediatric heart transplantation (HTx). We performed a retrospective chart review of 94 1-year HTx survivors, 1988-2006. Clinical records identified patients with arrhythmias occurring >1 year after HTx requiring pharmacotherapy, excluding acute rejection. We reviewed preoperative diagnosis, gender, age at HTx, operative details, transplant coronary artery disease (TCAD), and mortality. We analyzed serial ECGs after HTx for HR, PR, QRS, QT, and QTc intervals. Our results found complete data in 58 patients, 14 (24%) with arrhythmia and 44 controls. Arrhythmias occurred 1.1-17.9 years after HTx (mean = 6.8): 11 focal atrial tachycardia, 1 atrial fibrillation/flutter, 1 atrioventricular node reentry tachycardia; only 1 patient had ventricular tachycardia (VT). Serial ECG intervals were similar between groups, as well as surgical technique, ischemic time, and rejection history. Seven patients (50%) with arrhythmias had death or graft death versus 11% of the controls (P = 0.006). Patients with arrhythmias were more likely to be diagnosed with TCAD (P = 0.007). The patient with VT had no TCAD. In conclusion, supraventricular arrhythmias were frequent (22%) in 1-year survivors of pediatric HTx. These patients were more likely to develop TCAD and/or graft loss/mortality.


Use of oscillatory positive expiratory pressure (OPEP) devices to augment sputum clearance in COPD: a systematic review and meta-analysis.
Oscillating positive expiratory pressure (OPEP) devices are intended to facilitate sputum clearance in chronic obstructive pulmonary disease (COPD), but there is uncertainty as to their place in treatment pathways. We aimed to review the existing literature to establish the evidence base for their use. A systematic search of records up to March 2020 was performed on PubMed, CINAHL, Medline (Ovid), Cochrane and Embase to retrieve clinical trials that evaluated the efficacy of OPEP devices in patients with COPD. Two independent reviewers retrieved the titles, abstracts and full texts, and completed the data extraction. Following full-text review of 77 articles, eight (six randomised control trials and 2 cross-over studies) were eligible for inclusion. Pooled analysis showed low-grade evidence that the use of OPEP devices was associated with decreased COPD symptoms and exacerbations (OR 0.37, 95% CI 0.19 to 0.72), and enhanced exercise capacity; 6 min walk distance (mean difference (95% CI), 49.8 m (14.2 m to 85.5 m); p=0.009]). However, studies were mostly short term with the majority having a high risk of bias. The average acceptance, completion and drop-out rates were 82%, 91% and 8%, respectively. The use of OPEP devices can have a positive impact in COPD, but confidence in effect sizes is low and there is a need for further, higher quality studies to examine their long-term efficacy in COPD as well as to identify specific patient phenotypes that are more likely to respond. CRD 42016041835.


Implications of recombination for HIV diversity.
The human immunodeficiency virus (HIV) population is characterised by extensive genetic variability that results from high error and recombination rates of the reverse transcription process, and from the fast turnover of virions in HIV-infected individuals. Among the viral variants encountered at the global scale, recombinant forms are extremely abundant. Some of these recombinants (known as circulating recombinant forms) become fixed and undergo rapid expansion in the population. The reasons underlying their epidemiological success remain at present poorly understood and constitute a fascinating area for future research to improve our understanding of immune escape, pathogenicity and transmission. Recombinant viruses are generated during reverse transcription as a consequence of template switching between the two genetically different genomic RNAs present in a heterozygous virus. Recombination can thereby generate shortcuts in evolution by producing mosaic reverse transcription products of parental genomes. Therefore, in a single infectious cycle multiple mutations that are positively selected can be combined or, conversely, negatively selected mutations can be removed. Recombination is therefore involved in different aspects of HIV evolution, adaptation to its host, and escape from antiviral treatments.


HIV prevalence and behavioral risk factors in the Sudan People's Liberation Army: Data from South Sudan.
After two decades of civil war, South Sudan has limited published data on HIV prevalence and behavioral determinants of HIV infection risk. A surge in HIV/AIDS prevalence is a real concern for this new country with limited access to medical or HIV preventive services, and low education and literacy levels. We present findings from the first bio-behavioral surveillance survey conducted within the Sudan People's Liberation Army (SPLA). A cross-sectional survey of 1,149 randomly selected soldiers from thirteen SPLA bases was conducted in two phases: July to August 2010 and April to May 2012. Consenting participants received HIV rapid tests, pre- and post-test counseling, and a personal interview. Demographics, knowledge, attitudes, and behaviors, including sexual behavior, alcohol use, and mental health were assessed using computer-assisted interviews. The final sample included 1,063 survey participants (96.7% male). Education levels within the SPLA are low; only 16.4% attended school beyond the primary level. The overall HIV prevalence in the sample was 5.0% (95% confidence interval [CI]: 3.6-6.9). High-risk behaviors (e.g., multiple or concurrent sexual partners, heavy alcohol use, low condom use) were noted among SPLA members. High levels of HIV stigma were identified: 90.6% (n = 916) responded with one or more negative beliefs towards PLHIV, and 60.3% thought a healthy-looking person with HIV should not be allowed to remain in the SPLA. Results from this first evaluation of risk behaviors and HIV prevalence among the SPLA highlight high-risk behaviors that may contribute to the spread of HIV. Understanding potential comorbid conditions will be critical to designing strategies to reduce HIV risk. This survey represents the first steps in understanding the HIV epidemic within the SPLA context.


Development of a Gold Nanoparticle-Based Lateral-Flow Immunoassay for Pneumocystis Pneumonia Serological Diagnosis at Point-of-Care.
Pneumocystis jirovecii pneumonia (PcP) is a major human immunodeficiency virus (HIV)-related illness, rising among immunocompromised non-HIV patients and in developing countries. Presently, the diagnosis requires respiratory specimens obtained through invasive and costly techniques that are difficult to perform in all patients or implement in all economic settings. Therefore, the development of a faster, cost-effective, non-invasive and field-friendly test to diagnose PcP would be a significant advance. In this study, recombinant synthetic antigens (RSA) of P. jirovecii's major surface glycoprotein (Msg) and kexin-like serine protease (Kex1) were produced and purified. These RSA were applied as antigenic tools in immunoenzymatic assays for detection of specific anti-P. jirovecii antibodies (IgG and IgM) in sera of patients with (n = 48) and without (n = 28) PcP. Results showed that only IgM anti-P. jirovecii levels were significantly increased in patients with PcP compared with patients without P. jirovecii infection (p ≤ 0.001 with both RSA). Thus, two strip lateral flow immunoassays (LFIA), based on the detection of specific IgM anti-P. jirovecii antibodies in human sera samples, were developed using the innovative association of P. jirovecii's RSA with spherical gold nanoparticles (AuNPs). For that, alkanethiol-functionalized spherical AuNPs with ca. ~40 nm in diameter were synthetized and conjugated with the two RSA (Msg or Kex1) produced. These AuNP-RSA conjugates were characterized by agarose gel electrophoresis (AGE) and optimized to improve their ability to interact specifically with serum IgM anti-P. jirovecii antibodies. Finally, two LFIA prototypes were developed and tested with pools of sera from patients with (positive sample) and without (negative sample) PcP. Both LFIA had the expected performance, namely, the presence of a test and control red colored lines with the positive sample, and only a control red colored line with the negative sample. These results provide valuable insights into the possibility of PcP serodiagnosis at point-of-care. The optimization, validation and implementation of this strip-based approach may help to reduce the high cost of medical diagnosis and subsequent treatment of PcP both in industrialized and low-income regions, helping to manage the disease all around the world.


Magnetization transfer MR imaging correlation with histopathology in intracranial tuberculomas.
The purpose of this study was to correlate the differences in the magnetization transfer (MT) ratios of different components of the tuberculoma with histopathology and to see whether MT or conventional MR imaging correlates better with histopathology. MT T1 and conventional spin echo MR imaging was performed in six patients with intracranial tuberculomas. The tuberculomas were excised as a single mass and ex vivo MR imaging was performed using the same protocol. The gross histopathology was compared with in vivo imaging with respect to the MR signal intensity (MT ratio) in all six specimens. The size of the tuberculomas was larger on MT T1-weighted images compared to T2-weighted images and matched the gross measurements of each specimen. The MT hyperintense rim matched the cellular component of the tuberculoma that was masked on T2-weighted images because of the associated perifocal oedema. The cellular component had a lower MT ratio compared to the necrotic components. The outer hyperintense rim and hyperintense strands are due to the cellular infiltrate, noncaseating granulomas, and gliosis while the hypointense core represents solid caseation. The cellular outer rim shows lower MT ratio compared to the core of the tuberculoma. Histological correlation of the cellular and necrotic components of tuberculomas is best shown with MT T1 imaging.


A review of deep learning in medical imaging: Imaging traits, technology trends, case studies with progress highlights, and future promises.
Since its renaissance, deep learning has been widely used in various medical imaging tasks and has achieved remarkable success in many medical imaging applications, thereby propelling us into the so-called artificial intelligence (AI) era. It is known that the success of AI is mostly attributed to the availability of big data with annotations for a single task and the advances in high performance computing. However, medical imaging presents unique challenges that confront deep learning approaches. In this survey paper, we first present traits of medical imaging, highlight both clinical needs and technical challenges in medical imaging, and describe how emerging trends in deep learning are addressing these issues. We cover the topics of network architecture, sparse and noisy labels, federating learning, interpretability, uncertainty quantification, etc. Then, we present several case studies that are commonly found in clinical practice, including digital pathology and chest, brain, cardiovascular, and abdominal imaging. Rather than presenting an exhaustive literature survey, we instead describe some prominent research highlights related to these case study applications. We conclude with a discussion and presentation of promising future directions.


Stimulated parametric emission microscopy.
We propose a novel microscopy technique based on the four-wave mixing (FWM) process that is enhanced by two-photon electronic resonance induced by a pump pulse along with stimulated emission induced by a dump pulse. A Ti:sapphire laser and an optical parametric oscillator are used as light sources for the pump and dump pulses, respectively. We demonstrate that our proposed FWM technique can be used to obtain a one-dimensional image of ethanol-thinned Coumarin 120 solution sandwiched between a hole-slide glass and a cover slip, and a two-dimensional image of a leaf of Camellia sinensis.


A clinical test for superior glenoid labral or 'SLAP' lesions.
To describe a clinical test associated with unstable lesions of the superior glenoid labrum-long head biceps tendon origin, or SLAP (superior labrum anterior to posterior). Description of a newly discovered clinical sign that correlated with SLAP pathology. Retrospective review of 66 consecutive arthroscopically confirmed SLAP lesions to determine the sensitivity of the SLAPprehension test. Orthopedic sports medicine clinics with an emphasis on shoulder problems. Patients with shoulder pain and arthroscopically verified lesions of the superior glenoid labrum and conjoined long head biceps tendon. Shoulder arthroscopy and in some cases arthroscopic SLAP lesion repair. Nonapplicable. The SLAPprehension test involves cross chest adduction (horizontal flexion) of the affected shoulder with the elbow extended and forearm pronated. A positive maneuver produces either apprehension, pain referable to the bicipital groove, and an audible or palpable click. The test is repeated with the forearm supinated, which must cause diminution of the pain. Mechanically, elbow extension and forearm pronation places traction on the long head biceps tendon. When anterior scapular protraction is limited by the clavicle, further adduction entraps the unstable biceps tendon and superior glenoid labrum between the glenoid fossa and humeral head. Forearm supination decreases traction on the long head biceps tendon and allows for reduction of the unstable labrum complex with lessening of the pain. A retrospective chart review of 66 consecutive arthroscopically verified shoulders with SLAP lesions revealed the SLAPprehension test to be 87.5% sensitive for unstable SLAP lesions. The SLAPprehension test is helpful in the clinical evaluation of patients with unstable superior glenoid labrum lesions whose symptoms are often confused and overlap with those of shoulder impingement or acromioclavicular arthrosis.


Modeling Outcomes of First-Line Antiretroviral Therapy and Rate of CD4 Counts Change among a Cohort of HIV/AIDS Patients in Ethiopia: A Retrospective Cohort Study.
Antiretroviral therapy has shown to be effective in reducing morbidity and mortality in patients infected with HIV for the past couples of decades. However, there remains a need to better understand the characteristics of long-term treatment outcomes in resource poor settings. The main aim of this study was to determine and compare the long-term response of patients on nevirapine and efavirenz based first line antiretroviral therapy regimen in Ethiopia. Hospital based retrospective cohort study was conducted from January 2009 to December 2013 at University hospital located in Northwest Ethiopia. Human subject research approval for this study was received from University of Gondar Research Ethics Committee and the medical director of the hospital. Cox-proportional hazards model was used to assess the effect of baseline covariates on composite outcome and a semi-parametric mixed effect model was used to investigate CD4 counts response to treatments. A total of 2386 HIV/AIDS naive patients were included in this study. Nearly one-in-four patients experienced the events, of which death, lost to follow up, treatment substitution and discontinuation of Non-Nucleoside Reverse Transcriptase Inhibitors(NNRTI) accounted: 99 (26.8%), 122 (33.0%), 137 (37.0%) and 12 (3.2%), respectively. The hazard of composite outcome on nevirapine compared with efavirenz was 1.02(95%CI: 0.52-1.99) with p-value = 0.96. Similarly, the hazard of composite outcome on tenofovir and stavudine compared with zidovudine were 1.87 (95%CI: 1.52-2.32), p-value < 0.0001 and 1.72(95% CI: 1.22-2.32), p-value = 0.002, respectively. The rate of CD4 increase in response to treatment was high during the first 10 months and stabilized later. This study revealed that treatment responses were comparable whether nevirapine or efavirenz was chosen to initiate antiretroviral therapy for HIV/AIDS patients in Ethiopia. There was significant difference on risk of composite outcome between patients who were initiated with Tenofovir containing ART regimen compared with zidovudine after controlling for NNRTI drug combinations.


Persistence of Amygdala-Hippocampal Connectivity and Multi-Voxel Correlation Structures During Awake Rest After Fear Learning Predicts Long-Term Expression of Fear.
After encoding, memories undergo a process of consolidation that determines long-term retention. For conditioned fear, animal models postulate that consolidation involves reactivations of neuronal assemblies supporting fear learning during postlearning "offline" periods. However, no human studies to date have investigated such processes, particularly in relation to long-term expression of fear. We tested 24 participants using functional MRI on 2 consecutive days in a fear conditioning paradigm involving 1 habituation block, 2 acquisition blocks, and 2 extinction blocks on day 1, and 2 re-extinction blocks on day 2. Conditioning blocks were preceded and followed by 4.5-min rest blocks. Strength of spontaneous recovery of fear on day 2 served as a measure of long-term expression of fear. Amygdala connectivity primarily with hippocampus increased progressively during postacquisition and postextinction rest on day 1. Intraregional multi-voxel correlation structures within amygdala and hippocampus sampled during a block of differential fear conditioning furthermore persisted after fear learning. Critically, both these main findings were stronger in participants who exhibited spontaneous recovery 24 h later. Our findings indicate that neural circuits activated during fear conditioning exhibit persistent postlearning activity that may be functionally relevant in promoting consolidation of the fear memory.


The 1.38 A crystal structure of DmsD protein from Salmonella typhimurium, a proofreading chaperone on the Tat pathway.
The DmsD protein is necessary for the biogenesis of dimethyl sulphoxide (DMSO) reductase in many prokaryotes. It performs a critical chaperone function initiated through its binding to the twin-arginine signal peptide of DmsA, the catalytic subunit of DMSO reductase. Upon binding to DmsD, DmsA is translocated to the periplasm via the so-called twin-arginine translocation (Tat) pathway. Here we report the 1.38 A crystal structure of the protein DmsD from Salmonella typhimurium and compare it with a close functional homolog, TorD. DmsD has an all-alpha fold structure with a notable helical extension located at its N-terminus with two solvent exposed hydrophobic residues. A major difference between DmsD and TorD is that TorD structure is a domain-swapped dimer, while DmsD exists as a monomer. Nevertheless, these two proteins have a number of common features suggesting they function by using similar mechanisms. A possible signal peptide-binding site is proposed based on structural similarities. Computational analysis was used to identify a potential GTP binding pocket on similar surfaces of DmsD and TorD structures.


Omalizumab in severe allergic asthma inadequately controlled with standard therapy: a randomized trial.
Inhaled corticosteroids (ICS) and long-acting β(2)-agonists (LABAs) are recommended in patients with asthma that is not well-controlled; however, many patients continue to have inadequately controlled asthma despite this therapy. To evaluate the efficacy and safety of omalizumab in patients with inadequately controlled severe asthma who are receiving high-dose ICS and LABAs, with or without additional controller therapy. Prospective, multicenter, randomized, parallel-group, double-blind, placebo-controlled trial. (ClinicalTrials.gov registration number: NCT00314575). 193 investigational sites in the United States and 4 sites in Canada. 850 patients aged 12 to 75 years who had inadequately controlled asthma despite treatment with high-dose ICS plus LABAs, with or without other controllers. Omalizumab (n = 427) or placebo (n = 423) was added to existing medication regimens for 48 weeks. The primary end point was the rate of protocol-defined exacerbations over the study period. Secondary efficacy end points included the change from baseline to week 48 in mean daily number of puffs of albuterol, mean total asthma symptom score, and mean overall score on the standardized version of the Asthma Quality of Life Questionnaire (AQLQ[S]). Safety end points included the frequency and severity of treatment-emergent adverse events. During 48 weeks, the rate of protocol-defined asthma exacerbations was significantly reduced for omalizumab compared with placebo (0.66 vs. 0.88 per patient; P = 0.006), representing a 25% relative reduction (incidence rate ratio, 0.75 [95% CI, 0.61 to 0.92]). Omalizumab improved mean AQLQ(S) scores (0.29 point [CI, 0.15 to 0.43]), reduced mean daily albuterol puffs (-0.27 puff/d [CI, -0.49 to -0.04 puff/d]), and decreased mean asthma symptom score (-0.26 [CI, -0.42 to -0.10]) compared with placebo during the 48-week study period. The incidence of adverse events (80.4% vs. 79.5%) and serious adverse events (9.3% vs. 10.5%) were similar in the omalizumab and placebo groups, respectively. The results are limited by early patient discontinuation (20.8%). The study was not powered to detect rare safety events or the treatment effect in the oral corticosteroid subgroup. In this study, omalizumab provided additional clinical benefit for patients with severe allergic asthma that is inadequately controlled with high-dose ICS and LABA therapy. Genentech and Novartis Pharmaceuticals.


Time from symptom onset to treatment and outcomes after thrombolytic therapy. GUSTO-1 Investigators.
This study sought to examine the relations among patient characteristics, time to thrombolysis and outcomes in the international GUSTO-I trial. Studies have shown better left ventricular function and decreased infarct size as well as increased survival with earlier thrombolysis, but the relative benefits of various thrombolytic agents with earlier administration are uncertain. We evaluated the relations of baseline characteristics to three prospectively defined time variables: symptom onset to treatment, symptom onset to hospital arrival (presentation delay) and hospital arrival to treatment (treatment delay). We also examined the relations of delays to clinical outcomes and to the relative 30-day mortality benefit with accelerated tissue-type plasminogen activator (t-PA) versus streptokinase. Female, elderly, diabetic and hypertensive patients had longer delays at all stages. Previous infarction or bypass surgery was an additional risk factor for treatment delay. Early thrombolysis was associated with lower overall mortality rate (< 2 h, 5.5%; > 4 h, 9.0%), but no additional relative benefit resulted from earlier treatment with accelerated t-PA versus streptokinase (p = 0.38). Longer presentation and treatment delays were both associated with increased mortality rate (presentation delay < 1 h, 5.6% and > 4 h, 8.6%; treatment delay < 1 h, 5.4%, and > 90 min, 8.1%). As time to treatment increased, the incidence of recurrent ischemia or reinfarction decreased, but the rates of shock, heart failure and stroke increased. Earlier treatment resulted in better outcomes, regardless of thrombolytic strategy. Elderly, female and diabetic patients were treated later, adding to their already substantial risk.


Comparison of the Tissue Distribution of a Long-Circulating Glucagon-like Peptide-1 Agonist Determined by Positron Emission Tomography and Quantitative Whole-Body Autoradiography.
Positron emission tomography (PET) is a molecular imaging modality that enables non-invasive visualization of tracer distribution and pharmacology. Recently, peptides with long half-lives allowed once-a-week dosing of glucagon-like peptide-1 receptor (GLP-1R) agonists with therapeutic applications in diabetes and obesity. PET imaging for such long-lived peptides is hindered by the typically used short-lived radionuclides. Zirconium-89 (89Zr) emerged as a promising PET radionuclide with a sufficiently long half-life to be applied for biodistribution studies of long-circulating biomolecules. A comparison between the biodistribution profiles obtained via 89Zr-PET and the current standard, quantitative whole-body autoradiography (QWBA), will be valuable for the development of novel peptide drugs. We determined the PET biodistribution of a 89Zr-labeled acylated peptide agonist of GLP-1R and compared it to the profile obtained by QWBA using analogous tritiated tracers for up to 1 week after administration. The plasma metabolic profile was obtained and identification was done for the tritiated tracers. We found that, at early time points, the biodistribution profiles agreed between PET and QWBA. At the latertime points, the 89Zr tracer remained primarily trapped in the kidneys. The introduction of desferrioxamine (DFO) chelator reduced the peptide stability, and UPLC-MS analysis identified a circulating metabolite arising from DFO hydrolysis. Kidney accumulation of radiolabeled peptides and DFO metabolic instability may compromise biodistribution studies using 89Zr-PET to support the development of new biopharmaceuticals. PET and QWBA biodistribution data correlated well during the absorption phase, but new and more stable 89Zr chelators are needed for a more accurate description of the elimination phase.


Ozone-induced foliar damage and release of stress volatiles is highly dependent on stomatal openness and priming by low-level ozone exposure in Phaseolus vulgaris.
Acute ozone exposure triggers major emissions of volatile organic compounds (VOCs), but quantitatively, it is unclear how different ozone doses alter the start and the total amount of these emissions, and the induction rate of different stress volatiles. It is also unclear whether priming (i.e. pre-exposure to lower O3 concentrations) can modify the magnitude and kinetics of volatile emissions. We investigated photosynthetic characteristics and VOC emissions in Phaseolus vulgaris following acute ozone exposure (600 nmol mol-1 for 30 min) under illumination and in darkness and after priming with 200 nmol mol-1 O3 for 30 min. Methanol and lipoxygenase (LOX) pathway product emissions were induced rapidly, followed by moderate emissions of methyl salicylate (MeSA). Stomatal conductance prior to acute exposure was lower in darkness and after low O3 priming than in light and without priming. After low O3 priming, no MeSA and lower LOX emissions were detected under acute exposure. Overall, maximum emission rates and the total amount of emitted LOX products and methanol were quantitatively correlated with total stomatal ozone uptake. These results indicate that different stress volatiles scale differently with ozone dose and highlight the key role of stomatal conductance in controlling ozone uptake, leaf injury and volatile release.


Combination of high-sensitivity troponin I and N-terminal pro-B-type natriuretic peptide predicts future hospital admission for heart failure in high-risk hypertensive patients with preserved left ventricular ejection fraction.
Additional risk stratification may provide more aggressive and focalized preventive treatment to high-risk hypertensive patients according to the Japanese hypertension guidelines. We prospectively investigated the predictive value of high-sensitivity troponin I (hsTnI), both independently and in combination with N-terminal pro-B-type natriuretic peptide (NT-proBNP), for incident heart failure (HF) in high-risk hypertensive patients with preserved left ventricular ejection fraction (LVEF). Baseline hsTnI and NT-proBNP levels and echocardiography data were obtained for 493 Japanese hypertensive outpatients (mean age, 68.5 years) with LVEF ≥ 50%, no symptomatic HF, and at least one of the following comorbidities: stage 3-4 chronic kidney disease, diabetes mellitus, and stable coronary artery disease. During a mean follow-up period of 86.1 months, 44 HF admissions occurred, including 31 for HF with preserved ejection fraction (HFpEF) and 13 for HF with reduced ejection fraction (HFrEF; LVEF <50%). Both hsTnI (p < 0.01) and NT-proBNP (p < 0.005) levels were significant independent predictors of HF admission. Furthermore, when the patients were stratified into 4 groups according to increased hsTnI (≥highest tertile value of 10.6 pg/ml) and/or increased NT-proBNP (≥highest tertile value of 239.7 pg/ml), the adjusted relative risks for patients with increased levels of both biomarkers versus neither biomarker were 13.5 for HF admission (p < 0.0001), 9.45 for HFpEF (p = 0.0009), and 23.2 for HFrEF (p = 0.003). Finally, the combined use of hsTnI and NT-proBNP enhanced the C-index (p < 0.05), net reclassification improvement (p = 0.0001), and integrated discrimination improvement (p < 0.05) to a greater extent than that of any single biomarker. The combination of hsTnI and NT-proBNP, which are individually independently predictive of HF admission, could improve predictions of incident HF in high-risk hypertensive patients but could not predict future HF phenotypes.


The nature of the mutagenicity and carcinogenicity of nitrated, aromatic compounds in the environment.
Gaseous substances such as nitrogen dioxide (NO2) and sulfur dioxide (SO2) stimulate the process of nitration of polycyclic aromatic hydrocarbons, and the transformation products display a broad spectrum of mutagenicity, genotoxicity, and carcinogenicity. Bacterial mutation by nitroarenes is specific. Tetracyclic nitroarenes are thought to be the most mutagenic compounds in the Salmonella test system, and some are carcinogenic in rats and mice. Furthermore, it was found that the mutational nitroarenes produced mostly DNA damage, which is subject to recombination repair in the rec assay system using Bacillus subtilis. Nitroarenes in the environment seem to be ubiquitous; the majority of the compounds are emitted directly from diesel emissions, kerosene heaters, and gas and liquefied-gas burners or heaters. In nitroarenes induced during incomplete combustion, nitropyrene and nitrofluoranthene derivatives are the most important mutagens/carcinogens for determining the chronic toxicity of nitroarenes overall.


Effect of alpha thalassaemia on the rheology of homozygous sickle cell disease.
A study of rheological determinants (plasma viscosity, whole-blood viscosity, and erythrocyte deformability) was made in 24 matched pairs of patients with homozygous sickle cell disease, with and without homozygous alpha-thalassaemia 2. Patients with coexisting alpha-thalassaemia showed a significant increase in erythrocyte deformability measured as filtration of washed erythrocytes through 5 micron diameter pores and also as viscosity of whole blood at high shear rate (230s-1) and standard haematocrit (0.45). This rheological advantage may explain the beneficial effect of alpha-thalassaemia 2 on haematological parameters and clinical events in homozygous sickle cell disease.


A Novel Typing Method for Streptococcus pneumoniae Using Selected Surface Proteins.
The diverse pneumococcal diseases are associated with different pneumococcal lineages, or clonal complexes. Nevertheless, intra-clonal genomic variability, which influences pathogenicity, has been reported for surface virulence factors. These factors constitute the communication interface between the pathogen and its host and their corresponding genes are subjected to strong selective pressures affecting functionality and immunogenicity. First, the presence and allelic dispersion of 97 outer protein families were screened in 19 complete pneumococcal genomes. Seventeen families were deemed variable and were then examined in 216 draft genomes. This procedure allowed the generation of binary vectors with 17 positions and the classification of strains into surfotypes. They represent the outer protein subsets with the highest inter-strain discriminative power. A total of 116 non-redundant surfotypes were identified. Those sharing a critical number of common protein features were hierarchically clustered into 18 surfogroups. Most clonal complexes with comparable epidemiological characteristics belonged to the same or similar surfogroups. However, the very large CC156 clonal complex was dispersed over several surfogroups. In order to establish a relationship between surfogroup and pathogenicity, the surfotypes of 95 clinical isolates with different serogroup/serotype combinations were analyzed. We found a significant correlation between surfogroup and type of pathogenic behavior (primary invasive, opportunistic invasive, and non-invasive). We conclude that the virulent behavior of S. pneumoniae is related to the activity of collections of, rather than individual, surface virulence factors. Since surfotypes evolve faster than MLSTs and directly reflect virulence potential, this novel typing protocol is appropriate for the identification of emerging clones.


The meaning of measures: pitfalls in behavioral and developmental research.
This article considers a number of issues encountered in the selection and use of measures in research in the area of behavioral and developmental pediatrics. In addition to considering basic psychometric principles, it is argued that problems inherent in the developmental process, the lack of correspondence between study and standardization samples, and methodological factors affecting measurement may threaten the validity of individual measures and bias research outcome. A number of recommendations are made to counter the effects of these common measurement problems that indirectly result from the absence of a "gold standard" for measuring behavior and development.


Multinuclear zinc bisamidinate catalyzed asymmetric alkylation of α-ketoesters and its unique chemoselectivity.
The multinuclear Zn-bisamidinate catalyzed enantioselective addition of Et2Zn to α-ketoesters has been developed. The steric tuning of two amidinate units as well as multiple coordination on the Zn atoms play a key role in achieving high enantioselectivity (up to 98% ee) and unique chemoselectivity. The present catalyst exhibited the preferential alkylation of α-ketoesters even in the presence of aldehydes.


Experimental Investigation and Simplistic Geochemical Modeling of CO₂ Mineral Carbonation Using the Mount Tawai Peridotite.
In this work, the potential of CO₂ mineral carbonation of brucite (Mg(OH)2) derived from the Mount Tawai peridotite (forsterite based (Mg)₂SiO4) to produce thermodynamically stable magnesium carbonate (MgCO3) was evaluated. The effect of three main factors (reaction temperature, particle size, and water vapor) were investigated in a sequence of experiments consisting of aqueous acid leaching, evaporation to dryness of the slurry mass, and then gas-solid carbonation under pressurized CO2. The maximum amount of Mg converted to MgCO₃ is ~99%, which occurred at temperatures between 150 and 175 °C. It was also found that the reduction of particle size range from >200 to <75 µm enhanced the leaching rate significantly. In addition, the results showed the essential role of water vapor in promoting effective carbonation. By increasing water vapor concentration from 5 to 10 vol %, the mineral carbonation rate increased by 30%. This work has also numerically modeled the process by which CO₂ gas may be sequestered, by reaction with forsterite in the presence of moisture. In both experimental analysis and geochemical modeling, the results showed that the reaction is favored and of high yield; going almost to completion (within about one year) with the bulk of the carbon partitioning into magnesite and that very little remains in solution.


Management and outcome in 75 individuals with long-chain fatty acid oxidation defects: results from a workshop.
At present, long-chain fatty acid oxidation (FAO) defects are diagnosed in a number of countries by newborn screening using tandem mass spectrometry. In the majority of cases, affected newborns are asymptomatic at time of diagnosis and acute clinical presentations can be avoided by early preventive measures. Because evidence-based studies on management of long-chain FAO defects are lacking, we carried out a retrospective analysis of 75 patients from 18 metabolic centres in Germany, Switzerland, Austria and the Netherlands with special regard to treatment and disease outcome. Dietary treatment is effective in many patients and can prevent acute metabolic derangements and prevent or reverse severe long-term complications such as cardiomyopathy. However, 38% of patients with very long-chain acyl-CoA dehydrogenase (VLCAD) deficiency had intermittent muscle weakness and pain despite adhering to therapy. Seventy-six per cent of patients with disorders of the mitochondrial trifunctional protein (TFP)-complex including long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency, had long-term myopathic symptoms. Of these, 21% had irreversible peripheral neuropathy and 43% had retinopathy. The main principle of treatment was a fat-reduced and fat-modified diet. Fat restriction differed among patients with different enzyme defects and was strictest in disorders of the TFP-complex. Patients with a medium-chain fat-based diet received supplementation of essential long-chain fatty acids. l-Carnitine was supplemented in about half of the patients, but in none of the patients with VLCAD deficiency identified by newborn screening. In summary, in this cohort the treatment regimen was adapted to the severity of the underlying enzyme defect and thus differed among the group of long-chain FAO defects.


Managing fatigue after cancer treatment: development of RESTORE, a web-based resource to support self-management.
The aim of this study is to co-create an evidence-based and theoretically informed web-based intervention (RESTORE) designed to enhance self-efficacy to live with cancer-related fatigue (CRF) following primary cancer treatment. A nine-step process informed the development of the intervention: (1) review of empirical literature; (2) review of existing patient resources; (3) establish theoretical framework; (4) establish design team with expertise in web-based interventions, CRF and people affected by cancer; (5) develop prototype intervention; (6) user testing phase 1; (7) refinement of prototype; (8) user testing phase 2; and (9) develop final intervention. Key stakeholders made a critical contribution at every step of intervention development, and user testing, which involved an iterative process and resulted in the final intervention. The RESTORE intervention has five sessions; sessions 1 and 2 include an introduction to CRF and goal setting. Sessions 3-5 can be tailored to user preference and are designed to cover areas of life where CRF may have an impact: home and work life, personal relationships and emotional adjustment. It is feasible to systematically 'co-create' an evidence-based and theory-driven web-based self-management intervention to support cancer survivors living with the consequences of cancer and its treatment. This is the first account of the development of a web-based intervention to support self-efficacy to manage CRF. An exploratory trial to test the feasibility and acceptability of RESTORE is now warranted.


An electron microscope study of basophile substances of frozen-dried rat liver.
Small pieces of liver from rats subjected to different dietary regimes were fixed by freeze-drying, and postfixed by in vacuo heating and denaturation with alcohol. Specimens were digested with ribo- or deoxyribonuclease, and stained with gallocyanin-chromalum, azure II, the Feulgen procedure or alcoholic platinic tetrabromide. Some specimens were reserved as controls of the effects of enzyme treatment. Stained and unstained specimens were embedded in methacrylate and examined by light and electron microscopy. Basophilic and Feulgen-positive substances, after contact with watery reagents, were found by electron microscopy to exist as small dense granules embedded in a less dense homogeneous matrix, forming the walls of submicroscopic vacuoles. These granules were absent after digestion with nucleodepolymerases. In specimens (unstained, or stained with platinic tetrabromide) which had not passed through water, the dense (basophile) substances in nuclei and cytoplasm were found to exist, not as granules, but as ill defined submicroscopic concentrates which blended imperceptibly into the homogeneous matrix of the vacuolar walls. Objections to the use of stains for improving contrast conditions in electron microscopy of tissues are discussed, and it is concluded that the reagents do not necessarily produce the observed increases in contrast by selectively stabilizing certain structures. The concept of microsomes as pre-existing distinct morphological entities in intact (unhomogenized) cells is thought to be inconsistent with the distribution of basophile substances in frozen-dried liver.


SKALP/elafin is an inducible proteinase inhibitor in human epidermal keratinocytes.
Skin-derived antileukoproteinase (SKALP), otherwise known as elafin, is a recently discovered epidermal proteinase inhibitor with specificity for polymorphonuclear leukocyte (PMN)-derived elastase and proteinase-3; in addition to the proteinase-inhibiting domain, SKALP contains several transglutaminase substrate motifs. SKALP is virtually absent in normal human epidermis but is found in a number of inflammatory skin diseases, including psoriasis. Here we report the induction and processing of SKALP in vivo and in vitro. SKALP expression in vivo could be demonstrated following injury in normal human epidermis, using histology, western blotting, northern blotting and a functional assay. In vitro, SKALP expression was studied in conventional submerged keratinocyte culture systems and in keratinocytes cultured in an air-liquid interface model. Induction of SKALP activity in epidermis could be measured as early as 16 hours after skin injury; immunohistological examination showed that SKALP expression was confined to the outer layers of the stratum spinosum and the stratum granulosum. Northern blot analysis revealed a 0.8 kb transcript, both in vivo (psoriatic skin, injured skin) and in vitro (cultured keratinocytes). Western blot analysis showed that the major SKALP form in vivo was a low molecular mass fragment, containing the antiproteinase domain. In all cultures that were positive for SKALP, larger (8-10 kDa) forms of SKALP, containing the N-terminal transglutaminase substrate motifs in addition to the antiproteinase domain, were found. SKALP expression in cultured cells was found to be dependent on the system used. In a submerged culture system, SKALP could be induced by fetal calf serum.(ABSTRACT TRUNCATED AT 250 WORDS)


Enhanced corticobulbar excitability in chronic smokers during visual exposure to cigarette smoking cues.
Neuroimaging studies of chronic smokers report altered activity of several neural regions involved in the processing of rewarding outcomes. Neuroanatomical evidence suggests that these regions are directly connected to the tongue muscle through the corticobulbar pathways. Accordingly, we examined whether corticobulbar excitability might be considered a somatic marker for nicotine craving. We compared motor-evoked potential (MEP) amplitudes recorded from the tongue and the extensor carpi radialis (control muscle) of chronic smokers under drug withdrawal and intake conditions as well as a nonsmoker group. All participants were tested during passive exposure to pictures showing a smoking cue or a meaningless stimulus. In the intake condition, chronic smokers were asked to smoke a real cigarette (CSn: group 1) or a placebo (CSp: group 2). Results show that MEP amplitudes recorded from the tongues of participants in the CSn and CSp groups under the withdrawal condition were selectively enhanced during exposure to a smoking cue. However, this effect on tongue MEP amplitudes disappeared in the intake condition for both the CSn and CSp groups. Limitations include the fact that the study was conducted in 2 different laboratories, the small sample size, the absence of data on chronic smoker craving strength and the different tastes of the real and placebo cigarettes. These results suggest that, in chronic smokers, tongue muscle MEP amplitudes are sensitive to neural processes active under the physiological status of nicotine craving. This finding implicates a possible functional link between neural excitability of the corticobulbar pathway and the reward system in chronic smokers.


Are state patterns of smoking different for different racial/ethnic groups? an application of multilevel analysis.
Tobacco use research has often assumed "average" effects across place, race, and socioeconomic position. We explored and mapped the variation in smoking prevalence for racial/ethnic groups by gender and state after adjusting for demographic factors. We executed a cross-sectional, weighted, two-level multilevel multiple regression analysis (individuals in states), with current smoking as the outcome, using the 1995-1996 Current Population Survey Tobacco Use Supplement, for non-Hispanic (NH) whites, NH blacks, and Hispanics. We also calculated adjusted smoking prevalence, 95% confidence intervals, Spearman correlations, and state residual-based maps to examine state patterns. We found different smoking patterns for each racial group. Black women's smoking rates were markedly lower than the national subgroup rate in six clustered states in the deep South. Smoking rates for whites were higher than the subgroup national rate in several Great Lakes states, Texas, Nevada, and North Carolina. For white women, several rural Midwest states displayed lower-than-expected smoking rates (Idaho, Utah, South Dakota, and Nebraska). We documented positive correlations for smoking prevalence between men and women within each racial group, but not between racial groups, indicating a race-specific pattern of smoking. We found that state tobacco variables (taxation and agriculture) did not account for remaining state smoking variance after inclusion of demographic variables. Multilevel modeling may enhance surveillance of tobacco use patterns. Focusing on race-specific state smoking patterns may illuminate why racial/ethnic minority groups exhibit lower smoking prevalence compared to their white counterparts, by examining context of smoking that may be race-specific.


Host genetics influences on HIV type-1 disease.
HIV host genetic studies seek to describe as comprehensively as possible the effect of human genetic variation on the individual response to HIV type-1 (HIV-1) infection. Many associations between specific gene variants and HIV-1 disease outcomes have been reported over the past 15 years. Although most of them have yet to be confirmed or have been proven false-positives, the identification of several definitive genotype-phenotype associations has shed new light on HIV-1 pathogenesis. This review discusses these results in the context of the new genome-wide approaches that now make it possible to globally assess the influence of the host genome on HIV-1-related outcomes.


Identifying high-yield low-emission pathways for the cereal production in South Asia.
Increasing agricultural production to meet the growing demand for food whilst reducing agricultural greenhouse gas (GHG) emissions is the major challenge under the changing climate. To develop long-term policies that address these challenges, strategies are needed to identify high-yield low-emission pathways for particular agricultural production systems. In this paper, we used bio-physical and socio-economic models to analyze the impact of different management practices on crop yield and emissions in two contrasting agricultural production systems of the Indo-Gangetic Plain (IGP) of India. The result revealed the importance of considering both management and socio-economic factors in the development of high-yield low-emission pathways for cereal production systems. Nitrogen use rate and frequency of application, tillage and residue management and manure application significantly affected GHG emissions from the cereal systems. In addition, various socio-economic factors such as gender, level of education, training on climate change adaptation and mitigation and access to information significantly influenced the adoption of technologies contributing to high-yield low-emission pathways. We discussed the policy implications of these findings in the context of food security and climate change.


Imaging canopy temperature: shedding (thermal) light on ecosystem processes.
Canopy temperature Tcan is a key driver of plant function that emerges as a result of interacting biotic and abiotic processes and properties. However, understanding controls on Tcan and forecasting canopy responses to weather extremes and climate change are difficult due to sparse measurements of Tcan at appropriate spatial and temporal scales. Burgeoning observations of Tcan from thermal cameras enable evaluation of energy budget theory and better understanding of how environmental controls, leaf traits and canopy structure influence temperature patterns. The canopy scale is relevant for connecting to remote sensing and testing biosphere model predictions. We anticipate that future breakthroughs in understanding of ecosystem responses to climate change will result from multiscale observations of Tcan across a range of ecosystems.


Validation of an 'endovascular-first' approach to spinal dural arteriovenous fistulas: an intention-to-treat analysis.
Spinal dural arteriovenous fistulas (SDAVFs) require pretreatment angiography; embolization can be performed in the same session. To validate this approach, obliteration and morbidity rates of 'endovascular-first' (embolization and microsurgery in the case of embolization failures) must be compared with rates for 'microsurgery-first' (microsurgical ligation without attempted embolization) approaches. We reviewed our institutional database (January 1998-October 2015) for SDAVFs, performing an intention-to-treat analysis comparing endovascular-first and microsurgery-first approaches. A total of 71 patients underwent surgical and/or endovascular treatment for SDAVFs. All SDAVFs were ultimately occluded. Of 35 patients under consideration for an endovascular-first approach, radicular artery anatomy or anterior spinal artery embolization risk precluded attempting embolization in seven cases (20%). Among 28 patients undergoing embolization, angiographic non-opacification of the fistula was noted in 18 (64%). Fourteen patients had obliteration with excellent casting of the draining vein (50%) and did not undergo surgery. There were no significant differences in total complications (9% vs 11%; p=1.0) or permanent complications (3% vs 4%; p=1.0) after attempted endovascular and surgical treatment. Based on an intention-to-treat analysis, there were no significant differences in total complications (11% vs 14%; p=1.0), permanent complications (6% vs 3%; p=0.61), or the symptomatic resolution/improvement rate (80% vs 78%; p=1.0) between endovascular-first and microsurgery-first groups. Our results support attempted embolization of SDAVFs prior to consideration of microsurgery, allowing for a less invasive treatment option in the same session as diagnostic angiography.


Attenuation of compensatory right ventricular hypertrophy and heart failure following monocrotaline-induced pulmonary vascular injury by the Na+-H+ exchange inhibitor cariporide.
Pulmonary hypertension results in compensatory right ventricular (RV) hypertrophy. We studied the role of the Na+-H+ exchange (NHE) in the latter process by determining the effect of the NHE-1 inhibitor cariporide after monocrotaline-induced pulmonary artery injury. Sprague-Dawley rats received a control or cariporide diet for 7 days, at which time they were administered either monocrotaline (60 mg/kg) or its vehicle. Twenty-one days later, monocrotaline control, but not cariporide-fed animals, demonstrated increased RV weights and cell size of 65 and 52%, respectively. Monocrotaline alone significantly increased RV systolic pressure and end diastolic pressure by 70 and 94%, respectively, whereas corresponding values with cariporide were significantly reduced to 33 and 42%. Central venous pressure increased by 414% in control animals, which was significantly reduced by cariporide. Monocrotaline treatment produced a decrease in cardiac output of 28 and 8% in the absence or presence of cariporide (P < 0.05 between groups), respectively. Although body weights were significantly lower in both monocrotaline-treated groups compared with vehicle treatment, with cariporide the net gain in body weight was twice that seen in the monocrotaline-treated animals without cariporide. Monocrotaline also increased RV NHE-1 and atrial natriuretic peptide mRNA expression, which was abrogated by cariporide. Monocrotaline-induced myocardial necrosis, fibrosis, and mononuclear infiltration was completely prevented by cariporide. Cariporide had no effect on monocrotaline-induced pulmonary intimal wall thickening. Our results demonstrate that cariporide directly attenuates myocardial dysfunction after monocrotaline administration independent of pulmonary vascular effects. NHE-1 inhibition may represent an effective adjunctive therapy that selectively targets myocardial hypertrophic responses in pulmonary vascular injury.


4-D generative model for PET/MRI reconstruction.
We introduce a 4-dimensional joint generative probabilistic model for estimation of activity in a PET/MRI imaging system. The model is based on a mixture of Gaussians, relating time dependent activity and MRI image intensity to a hidden static variable, allowing one to estimate jointly activity, the parameters that capture the interdependence of the two images and motion parameters. An iterative algorithm for optimisation of the model is described. Noisy simulation data, modeling 3-D patient head movements, is obtained with realistic PET and MRI simulators and with a brain phantom from the BrainWeb database. Joint estimation of activity and motion parameters within the same framework allows us to use information from the MRI images to improve the activity estimate in terms of noise and recovery.


Economic impacts of the Pueblo Smoke-Free Air Act.
On July 1, 2003, the city of Pueblo CO enacted a smokefree ordinance (Pueblo Smoke-Free Air Act [PSFAA]) that prohibited smoking in public places and workplaces, including taverns and restaurants. Opponents to this ordinance argued that it would have a negative impact on tavern and restaurant sales. The purpose of this study was to determine whether the PSFAA had a negative economic impact on tavern and restaurant sales tax revenues. With data gathered in 2007, this study implemented an interrupted time-series model in 2008, using 42 months of pre- and post-intervention sales tax revenue data for Pueblo to determine whether implementation of this ordinance had an effect on tavern and restaurant sales tax revenues. Ratios of tavern and restaurant openings to closings were also computed for the pre- and post-intervention periods. Pre-post sales tax revenue data showed slight losses in sales tax revenue for taverns, and gains for restaurants, which more than offset the tavern losses. After adjusting for the consumer price index, the city of Pueblo experienced a 20.3% gain in combined tavern and restaurant sales tax revenues from the pre-ordinance period to the post-ordinance period. The ratio of tavern openings to closings improved from 1:1 pre-period to 3.3:1 post-period and the restaurant ratio remained unchanged at approximately 1.78:1 from pre- to post-period. There is no evidence that the PSFAA had a negative economic impact on consumer price index-adjusted tavern and restaurant sales tax revenues. From a fiscal policy perspective, this ordinance may have contributed to a net increase in sales tax revenues for the city of Pueblo. The business openings/closings data suggest that the confidence Pueblo's business sector had in the local hospitality industry was not negatively influenced by the PSFAA.


How Oxygen Exposure Improves the Back Contact and Performance of Antimony Selenide Solar Cells.
The improvement of antimony selenide solar cells by short-term air exposure is explained using complementary cell and material studies. We demonstrate that exposure to air yields a relative efficiency improvement of n-type Sb2Se3 solar cells of ca. 10% by oxidation of the back surface and a reduction in the back contact barrier height (measured by J-V-T) from 320 to 280 meV. X-ray photoelectron spectroscopy (XPS) measurements of the back surface reveal that during 5 days in air, Sb2O3 content at the sample surface increased by 27%, leaving a more Se-rich Sb2Se3 film along with a 4% increase in elemental Se. Conversely, exposure to 5 days of vacuum resulted in a loss of Se from the Sb2Se3 film, which increased the back contact barrier height to 370 meV. Inclusion of a thermally evaporated thin film of Sb2O3 and Se at the back of the Sb2Se3 absorber achieved a peak solar cell efficiency of 5.87%. These results demonstrate the importance of a Se-rich back surface for high-efficiency devices and the positive effects of an ultrathin antimony oxide layer. This study reveals a possible role of back contact etching in exposing a beneficial back surface and provides a route to increasing device efficiency.


Divergent evolution peaks under intermediate population bottlenecks during bacterial experimental evolution.
There is growing evidence that parallel molecular evolution is common, but its causes remain poorly understood. Demographic parameters such as population bottlenecks are predicted to be major determinants of parallelism. Here, we test the hypothesis that bottleneck intensity shapes parallel evolution by elucidating the genomic basis of adaptation to antibiotic-supplemented media in hundreds of populations of the bacterium Pseudomonas fluorescens Pf0-1. As expected, bottlenecking decreased the rate of phenotypic and molecular adaptation. Surprisingly, bottlenecking had no impact on the likelihood of parallel adaptive molecular evolution at a genome-wide scale. However, bottlenecking had a profound impact on the genes involved in antibiotic resistance. Specifically, under either intense or weak bottlenecking, resistance predominantly evolved by strongly beneficial mutations which provide high levels of antibiotic resistance. In contrast with intermediate bottlenecking regimes, resistance evolved by a greater diversity of genetic mechanisms, significantly reducing the observed levels of parallel genetic evolution. Our results demonstrate that population bottlenecking can be a major predictor of parallel evolution, but precisely how may be more complex than many simple theoretical predictions.


Constitutive RAC activation is not sufficient to initiate melanocyte neoplasia but accelerates malignant progression.
Deregulated Ras signaling initiates and maintains melanocyte neoplasia. The Rho-like GTPase Rac has been implicated in Ras-induced neoplastic transformation. Moreover, a recurrent UV-induced mutation activating RAC1 has recently been detected in human melanoma. Here, a role for Rac in melanoma initiation and progression was investigated in human melanomas and zebrafish models of melanocyte neoplasia. Immunohistochemical analysis revealed RAC expression and activity restricted to melanocytes at the junction of the epidermis and dermis in benign neoplasms. Malignant melanocytes displayed elevated RAC activity that extended into the suprabasal epidermis, deeper into the dermis, and was maintained in metastases. Previously, we have used zebrafish transgenic models to demonstrate that deregulated Ras/Raf/mitogen-activated protein kinase signaling can initiate melanocyte neoplasia. Expression of a constitutively active RAC1 mutant (V12RAC1) was not sufficient to initiate melanocyte neoplasia in this organism. Furthermore, we did not detect an additive effect when combined with V600EBRAF, nor could V12RAC1 substitute for suppressed Pi3k signaling to restore melanoma progression. However, coexpression of V12RAC1 and oncogenic RAS accelerated tumor nodule formation. Immunohistochemical analysis revealed that the Rac activator Tiam1 (T-cell lymphoma invasion and metastasis 1) is overexpressed in melanoma tumor nodules in both zebrafish and humans. Thus, our data suggest that Rac contributes to the progression of melanoma and that Tiam1 may activate Rac in nodular presentations.


Toward Anion-π Interactions Directed Self-Assembly with Predesigned Dual Macrocyclic Receptors and Dianions.
Realizing anion-π interaction induced self-assembly with charge-neutral π receptors as building components is extremely challenging. We designed and synthesized a series of bisoxacalix[2]arene[2]triazines 7-11 in which two macrocyclic motifs are linked in diverse branching angle and rigidity. Crystal structures showed the use of rigid linkers is able to control the orientation of the two macrocyclic cavities. The interplay between the two cavities was revealed by binding studies of 8-11 with chloride in solution. Whereas 180°- and 120°-branched hosts 8 and 9 possess dual complexation ability, 60°-branched and flexibly linked hosts 10 and 11 only form 1:1 complex with chloride. Association and self-assembly of these bismacrocyclic building units with dianionic naphthalene-1,5-disulfonate were systematically investigated. The formation of oligomeric self-assemblies and large aggregates in solution was suggested by 1H NMR titrations, concentration- and temperature-variable 1H NMR, diffusion-ordered spectroscopy (DOSY), ESI-MS, and dynamic light scattering (DLS). The anion-π induced long-distance self-assembly with coherent particle formation was revealed by SEM, TEM, cryo-TEM, and AFM techniques. The systematic studies allowed us to draw the conclusion that the dianion served to bridge the initial host aggregates through anion-π interactions and was responsible for the coherent particle formation. The cavity orientation of the bismacrocycle components was found to have a significant influence on the coherent particle morphology.


Fabrication of dual stimuli-responsive multicompartmental drug carriers for tumor-selective drug release.
There has been increasing attention to the development of multi-stimuli-responsive drug carriers for precisely controlled drug release at target disease areas. In this study, pH- and redox-responsive hybrid drug carriers were fabricated by using both ketal-based acid-cleavable precursors and disulfide-based reducible precursors via stop-flow lithography. pH- and redox-sensitive drug release of the dual stimuli-responsive hybrid particles was confirmed, demonstrating their feasibility for selective and efficient drug release into tumor tissues in acidic and highly reductive environments. It was also found that the drug release rate of the particles was fine-tuned by modulating monomer compositions in the precursor. Importantly, the dual stimuli-responsive hybrid particles exhibited synergistic, controlled drug release in complex stimuli (both pH and redox stimuli) environments. To achieve tumor-selective combination chemotherapy, multicompartmental drug carriers consist of an acid-degradable compartment and a reducible compartment, which can separately encapsulate individual model drugs in each of the compartments. The multicompartmental particles exhibited independent drug release upon exposure to the corresponding stimulus. The dual stimuli-responsive, multicompartmental particles are effective drug carriers for tumor-selective release of a drug cocktail, leading to synergistic combination chemotherapy.


Low sensitivity in clinical diagnoses of dementia with Lewy bodies.
The success of future neurodegenerative disease (ND) therapies depends partly on accurate antemortem diagnoses. Relatively few prior studies have been performed on large, multicenter-derived datasets to test the accuracy of final clinical ND diagnoses in relation to definitive neuropathological findings. Data were analyzed from the University of Kentucky Alzheimer's Disease Center autopsy series and from the National Alzheimer's Coordinating Center (NACC) registry. NACC data are derived from 31 different academic medical centers, each with strong clinical expertise and infrastructure pertaining to NDs. The final clinical diagnoses were compared systematically with subsequent neuropathology diagnoses. Among subjects meeting final inclusion criteria (N = 2,861 for NACC Registry data), the strength of the associations between clinical diagnoses and subsequent ND diagnoses was only moderate. This was particularly true in the case of dementia with Lewy bodies (DLB): the sensitivity of clinical diagnoses was quite low (32.1% for pure DLB and 12.1% for Alzheimer's disease (AD + DLB) although specificity was over 95%. AD clinical diagnoses were more accurate (85.0% sensitivity and 51.1% specificity). The accuracy of clinical DLB diagnoses became somewhat lower over the past decade, due apparently to increased "over-calling" the diagnosis in patients with severe cognitive impairment. Furthermore, using visual hallucinations, extrapyramidal signs, and/or fluctuating cognition as part of the clinical criteria for DLB diagnosis was of minimal utility in a group (N = 237) with high prevalence of severe dementia. Our data suggest that further work is needed to refine our ability to identify specific aging-related brain disease mechanisms, especially in DLB.


Detection of West Nile virus-infected mosquitoes and seropositive juvenile birds in the vicinity of virus-positive dead birds.
Mosquitoes and wild birds were collected from three sites near locations in the New York City metropolitan area where single, West Nile (WN) virus-positive dead birds were found early in the 2000 transmission season. The mosquitoes were tested for the presence of infectious virus with a Vero cell culture assay and for WN viral RNA by using reverse transcriptase-polymerase chain reaction (RT-PCR) protocols. Serum samples from wild birds were tested for the presence of neutralizing antibodies against WN virus. Infectious WN virus and WN viral RNA were found in Culex species adult mosquitoes from each of the three sites, and a seropositive hatch-year house sparrow (Passer domesticus) was found in one of the three sites. Molecular techniques used to identify the species in the positive mosquito pools found that most of the pools contained a combination of Culex pipiens and Cx. restuans. The minimum infection rate in Culex species mosquitoes from the sites ranged from 0.2 to 6.0 per 1,000 specimens tested. The results demonstrated that, at least early in the transmission season, detection of a WN virus-positive dead bird indicates a local WN virus transmission cycle. This information is valuable in focusing subsequent surveillance and vector management programs. In addition, the RT-PCR procedure for detecting WN viral RNA in mosquito pools detected more positive pools than did the Vero cell plaque assay.


Towards wireless highly sensitive capacitive strain sensors based on gold colloidal nanoparticles.
We designed, produced and characterized new capacitive strain sensors based on colloidal gold nanoparticles. The active area of these sensors, made up of a 1 mm2 close-packed assembly of gold nanoparticles between interdigitated electrodes, was designed to achieve measurable capacitance (>∼1 pF) and overcome parasitic capacitances. Electro-mechanical experiments revealed that the sensitivity of such capacitive sensors increases in relation to the size of the nanoparticles. In the case of 14 nm gold NPs, such sensors present a relative capacitance variation of -5.2% for a strain of 1.5%, which is more than 5 times higher than that observed for conventional capacitive strain gauges. The existence of two domains (pure capacitive domain and mixed capacitive-resistance domain) as a function of the frequency measurement allows for the adaptation of sensitivity of these capacitive sensors. A simple low-cost circuit based on a microcontroller board was finally developed to detect the capacitance variations of such NP based strain sensors. This low-cost equipment paves the way for the development of an entirely wireless application set-up.


Anchor-based classification and type-C inhibitors for tyrosine kinases.
Tyrosine kinases regulate various biological processes and are drug targets for cancers. At present, the design of selective and anti-resistant inhibitors of kinases is an emergent task. Here, we inferred specific site-moiety maps containing two specific anchors to uncover a new binding pocket in the C-terminal hinge region by docking 4,680 kinase inhibitors into 51 protein kinases, and this finding provides an opportunity for the development of kinase inhibitors with high selectivity and anti-drug resistance. We present an anchor-based classification for tyrosine kinases and discover two type-C inhibitors, namely rosmarinic acid (RA) and EGCG, which occupy two and one specific anchors, respectively, by screening 118,759 natural compounds. Our profiling reveals that RA and EGCG selectively inhibit 3% (EGFR and SYK) and 14% of 64 kinases, respectively. According to the guide of our anchor model, we synthesized three RA derivatives with better potency. These type-C inhibitors are able to maintain activities for drug-resistant EGFR and decrease the invasion ability of breast cancer cells. Our results show that the type-C inhibitors occupying a new pocket are promising for cancer treatments due to their kinase selectivity and anti-drug resistance.


Thoracodorsal vessels as recipient vessels for the free TRAM flap in delayed breast reconstruction.
The internal mammary vessels have been recommended as the first choice recipient vessels for delayed breast reconstruction with the free TRAM flap. This approach has avoided surgery in the previously operated axilla, has required a shorter pedicle length, and has allowed for more medial placement of the TRAM tissue. Frequency of nonusable axillary vessels has been reported at 11 percent, with a 6 percent incidence of flap loss in the delayed reconstructive setting. We reviewed our experience with the thoracodorsal vessels as recipient vessels in delayed free TRAM breast reconstruction to assess more accurately the adequacy of these potential recipient vessels. All patients undergoing delayed TRAM reconstruction were reviewed. Forty-seven of 300 consecutive TRAM procedures were for planned delayed free reconstruction. In seven of the patients (15 percent), the thoracodorsal vessels were found to be inadequate for free reconstruction. A supercharged pedicled TRAM was used for reconstruction in each of these seven patients. Average operating room time was 7 hours. Mean follow-up time was 38 months. Nineteen percent of all patients developed at least one complication. Twelve percent of free TRAM patients developed a complication, whereas 57 percent of supercharged patients developed a postoperative complication. The difference in complication rates was statistically significant. The thoracodorsal vessels have provided an adequate recipient vessel in 85 percent of delayed free TRAM reconstructions, comparable to previous reports. Pedicling and supercharging the flap, in those situations in which the thoracodorsal vessels were inadequate, were associated with an increased incidence of postoperative complications. This suggests that in the delayed reconstructive setting, higher-risk patients benefit from free reconstruction over supercharged reconstructions. A second recipient vessel should be used when the thoracodorsal vessels are inadequate for planned free TRAM reconstruction. In these circumstances, we would recommend the use of the internal mammary vessels followed by the thoracoacromial vessels as reliable alternative recipient sites for delayed free TRAM reconstruction.


Rapid Low-Cost Microarray-Based Genotyping for Genetic Screening in Primary Immunodeficiency.
Background: Genetic tests for primary immunodeficiency disorders (PIDs) are expensive, time-consuming, and not easily accessible in developing countries. Therefore, we studied the feasibility of a customized single nucleotide variant (SNV) microarray that we developed to detect disease-causing variants and copy number variation (CNV) in patients with PIDs for only 40 Euros. Methods: Probes were custom-designed to genotype 9,415 variants of 277 PID-related genes, and were added to the genome-wide Illumina Global Screening Array (GSA). Data analysis of GSA was performed using Illumina GenomeStudio 2.0, Biodiscovery Nexus 10.0, and R-3.4.4 software. Validation of genotype calling was performed by comparing the GSA with whole-genome sequencing (WGS) data of 56 non-PID controls. DNA samples of 95 clinically diagnosed PID patients, of which 60 patients (63%) had a genetically established diagnosis (by Next-Generation Sequencing (NGS) PID panels or Sanger sequencing), were analyzed to test the performance of the GSA. The additional SNVs detected by GSA were validated by Sanger sequencing. Results: Genotype calling of the customized array had an accuracy rate of 99.7%. The sensitivity for detecting rare PID variants was high (87%). The single sample replication in two runs was high (94.9%). The customized GSA was able to generate a genetic diagnosis in 37 out of 95 patients (39%). These 37 patients included 29 patients in whom the genetic variants were confirmed by conventional methods (26 patients by SNV and 3 by CNV analysis), while in 8 patients a new genetic diagnosis was established (6 patients by SNV and 2 patients suspected for leukemia by CNV analysis). Twenty-eight patients could not be detected due to the limited coverage of the custom probes. However, the diagnostic yield can potentially be increased when newly updated variants are added. Conclusion: Our robust customized GSA seems to be a promising first-line rapid screening tool for PIDs at an affordable price, which opens opportunities for low-cost genetic testing in developing countries. The technique is scalable, allows numerous new genetic variants to be added, and offers the potential for genetic testing not only in PIDs, but also in many other genetic diseases.


Studies of human antiviral CD8+ lymphocytes using class I peptide tetramers.
Understanding the interactions between a host and a pathogen relies crucially on quantitative measurements of immune responses. Until recently, measurements of the levels of cellular immune responses, i.e. those mediated by CD4+ and CD8+ T lymphocytes have depended largely on culture in vitro and subsequent measurement of specific functions (such as cytolysis). More recently, new technologies based around tetrameric class I peptide complexes (tetramers) have allowed immunologists to measure CD8+ T lymphocyte levels directly ex vivo and independently of function. Since CD8+ lymphocytes play a key role in a number of important human viral infections, these tools have yielded useful insights into the dynamics, phenotype and function of human antiviral lymphocyte populations. In this review we describe some of the basic aspects of the biology of virus-specific CD8+ lymphocytes, and the current methods available to detect them. The use of tetramers has, in just four years, transformed our understanding of the immune responses against HIV, HTLV-1, HBV, HCV, CMV and EBV, and holds promise in a number of areas where quantitative analysis of the antiviral response in terms of both number and function is critical.


Angiogenesis in atherosclerotic plaque obtained from carotid endarterectomy: association between symptomatology and plaque morphology.
Carotid plaque with hemorrhage leads to cerebral embolism and ischemic stroke. Plaque angiogenesis and angiogenetic factors such as vascular endothelial growth factor (VEGF) are critical in the progression of atherosclerotic carotid plaque and intraplaque hemorrhage. The correlation between plaque angiogenesis and presence of clinical symptoms was studied in 41 specimens obtained during carotid endarterectomy from 20 symptomatic and 21 asymptomatic patients treated for carotid artery stenosis. Histological findings using hematoxylin-eosin and immunohistochemical staining against von Willebrand factor and VEGF were examined. Intraplaque hemorrhage, calcification, necrosis, and invasion of foam cells were frequently observed in the carotid plaques from symptomatic patients compared with asymptomatic patients. Higher microvessel density was found in the carotid plaques with necrosis and invasion of foam cells compared with plaques without necrosis and/or foam cell invasion, and higher expression of VEGF was found from symptomatic patients compared with asymptomatic patents. These results suggest that plaque angiogenesis and higher level of VEGF expression may enhance the progression of ischemic symptoms in patients with carotid artery stenosis. Invasive macrophages in the plaque of symptomatic patients increase levels of VEGF and might enhance plaque angiogenesis and atherosclerosis progression.


Results of complex rehabilitation patients with temporomandibular joint disease and parafunction of masticatory muscles.
Definition of the effective and appropriate essential use of myogymnastic exercises and orthopedic methods of posture correction in the complex rehabilitation of patients with TMJ pathology and parafunction of the masticatory muscles. In the study, 63 patients (11 men and 52 women) aged from 20 to 67, with the middle age 31±2.3 were treated, 43 of them had TMJ disease. In 51 patients, parafunctions of masticatory muscles were diagnosed. The treatment was carried out with the help of uncoupling the dentition rows of the splint, made after the TENS-therapy; used medicamentation treatment, using an individual plan for myogymnastic exercises and individual insoles for correction the posture. Total effectiveness of the treatment is shown as the average of the findings. Analysis of treatment dynamics was due to only those data the results of which after treatment in the control and main groups were different. Treatment effectiveness of the main group in comparison with the control one is as follows - periodontal vessels dopplerography - 16.22%, tonus of the right chewing muscle - 11.45%, right temporal muscle tone - 4.8%, the number of hours per a day in case of increased tone of the chewing muscles - 12.9%, mouth opening amplitude - 17.1%. Thus, total treatment effectiveness in the main group was 12.49±2.18% higher than in the control group. Results of complex rehabilitation, including myorelaxation procedures, medicamentation treatment, splint therapy, an individual plan of myogymnastic exercises for chewing muscles, and correction of posture using individual insoles showed its effectiveness more than 12.49±2.18% in comparison with traditional methods of treatment.


Distinct phenotypes of congenital acetylcholine receptor deficiency.
We contrast the phenotypes associated with hereditary acetylcholine receptor deficiency arising from mutations in either the acetylcholine receptor epsilon subunit or the endplate acetylcholine receptor clustering protein rapsyn. Mutational screening was performed by amplification of promoter and coding regions by PCR and direct DNA sequencing. We identified mutations in 37 acetylcholine receptor deficiency patients; 18 had acetylcholine receptor-epsilon mutations, 19 had rapsyn mutations. Mutated acetylcholine receptor-epsilon associated with bulbar symptoms, ptosis and ophthalmoplegia at birth, and generalized weakness. Mutated rapsyn caused either an early onset (rapsyn-EO) or late onset (rapsyn-LO) phenotype. Rapsyn-EO associated with arthrogryposis and life-threatening exacerbations during early childhood. Rapsyn-LO presented with limb weakness in adolescence or adulthood resembling seronegative myasthenia gravis. Awareness of distinct phenotypic features of acetylcholine receptor deficiency resulting from acetylcholine receptor-epsilon or rapsyn mutations should facilitate targeted genetic diagnosis, avoid inappropriate immunological therapy and, in some infants, prompt the rapid introduction of treatment that could be life saving.


[Sports and physical load bearing capacity after spondylodesis in patients with Scheuermann kyphosis].
In this investigation it was examined which is the effect of an extensive spondylodesis by patients with Scheuermann's kyphosis on their sport activity and their physical load capacity. Between 1983 and 1990, 10 patients with severe kyphosis secondary to Scheuermann's disease underwent surgical correction and spinal fusion. In 1997 all 10 patients were clinically and radiologically examined and interviewed at an average follow-up of 9.6 years. At the time of follow-up one patient was active in top sports, seven patients in normal sport, two patients were members in a sports-club and one patient was not interested in sport. Eight patients went in for sports regulary. Mostly the patients were active in gymnastics, cycling and swimming. At follow-up almost all patients found that their physical load capacity and the behaviour in relation to back pain were better than before operation. Half of the patients were exposed to a middle physical load in the daily life. The presented study shows that patients after extensive spondylodesis for Scheuermann's kyphosis and clear reduction of pain are not automatically limited in the daily life, both for sport and professional activity. However, generalized recommendations for the physical load capacity of the operated patient cannot be given, and an individualized decision must be taken for each case.


Blueberry Anthocyanin-Enriched Extracts Attenuate Fine Particulate Matter (PM2.5)-Induced Cardiovascular Dysfunction.
Blueberry anthocyanin-enriched extracts (BAE) at three doses (0.5, 1.0, and 2.0 g/kg) were administered by oral gavage to rats exposed to 10 mg/kg fine particulate matter (PM2.5) three times a week. A positive control group was exposed to PM2.5 without BAE treatment. We analyzed heart rate (HR), electrocardiogram (ECG), and histopathology, and biomarkers of cardiovascular system injuries, systemic inflammation, oxidative stress, endothelial function, and apoptosis. Results indicated that BAE, particularly at 1.0 g/kg, improved ECG and decreased cytokine levels in PM2.5-exposed rats. These changes were accompanied by an increase in interleukin 10 levels and superoxide dismutase activity in heart tissue and Bcl-2 protein expression, as well as a decrease in interleukin 6, malondialdehyde, endothelin 1, and angiotensin II levels and a reduction in Bax protein expression. This study demonstrates that BAE at certain doses can protect the cardiovascular system from PM2.5-induced damage.