Data integration for prediction of weight loss in randomized controlled dietary trials.
Diet is an important component in weight management strategies, but heterogeneous responses to the same diet make it difficult to foresee individual weight-loss outcomes. Omics-based technologies now allow for analysis of multiple factors for weight loss prediction at the individual level. Here, we classify weight loss responders (N = 106) and non-responders (N = 97) of overweight non-diabetic middle-aged Danes to two earlier reported dietary trials over 8 weeks. Random forest models integrated gut microbiome, host genetics, urine metabolome, measures of physiology and anthropometrics measured prior to any dietary intervention to identify individual predisposing features of weight loss in combination with diet. The most predictive models for weight loss included features of diet, gut bacterial species and urine metabolites (ROC-AUC: 0.84-0.88) compared to a diet-only model (ROC-AUC: 0.62). A model ensemble integrating multi-omics identified 64% of the non-responders with 80% confidence. Such models will be useful to assist in selecting appropriate weight management strategies, as individual predisposition to diet response varies.


Children's and parents' perceptions concerning surgical attire: a systematic review.
To review the literature about children's and parent's perceptions on surgical attire. A systematic search was conducted in the databases EMBASE, Latin American and Caribbean Health Sciences (LILACS), PubMed, PsycINFO, Scopus and Web of Science. Grey literature was searched on Google Scholar, Open Grey and ProQuest Dissertations, and Theses Database. A total of 2,567 papers were identified. After a two-phase selection, 15 studies were included in narrative synthesis. Children favored wearing white coats in five of the nine included studies (55.5% [95%CI 48.3-62.7]; p=1.00). With respect to parents' preferences, results of vote counting showed that in 11 of 15 included studies, they favored physicians wearing white coats (73.3% [95%CI 67.9-78.6]; p=0.11). Children and parents have preferred physicians to wear a white coat with a very low certainty of evidence.


A correlated histochemical and electron microscopic study of the intranuclear crystalline aggregates of adenovirus (RI-APC virus) in HeLa cells.
HeLa cells in tissue cultures infected with types 3, 4, or 7 of adenovirus (RI-APC virus) were studied in order to correlate certain histochemical and electron microscopic findings. Adjacent thin (ca. 0.05 micro) and thick (2-4 micro) sections of osmium-fixed, methacrylate-embedded cells were cut; by mapping the sections the same cells could be identified with both the electron and the light microscope. Intranuclear crystalline aggregates seen with the electron microscope to be composed of ordered arrays of viral particles were found by means of the Feulgen reaction to contain DNA. DNA is therefore assumed to be a constituent of the viral particle. The virus appeared to develop from an osmiophilic Feulgen-negative matrix. Displacement of nuclear chromatin occurred during this process. A Feulgen-azure staining method was found to permit clear distinction between viral and nuclear (host) DNA in thick sections.


Regular dilatation is unnecessary after tubularized incised-plate hypospadias repair.
To determine if patients who undergo tubularized incised-plate (TIP) urethroplasty need regular dilatation to prevent neourethral strictures or meatal stenosis. The follow-up of two consecutive series of patients who underwent primary TIP hypospadias repair by one surgeon was reviewed. The first 72 patients (group 1) had periodic neourethral calibrations for 1 year after repair, while the next 62 patients (group 2) had no routine dilatation, except for six patients (10%) in whom the meatus appeared to be small. Urethroscopy was performed in both groups at the time of fistula repair, skin revisions or during anaesthesia for unrelated procedures. There was no evidence of scarring in group 1; one case of meatal stenosis and one neourethral stricture were detected in group 2. The difference in outcome was not statistically significant (P = 0.4). Overall, the incidence of meatal stenosis was low (0.7%) and comparable with most recent series in which postoperative calibration was not used routinely. Dilatation of the neourethra is unnecessary after TIP urethroplasty. Calibration or uroflowmetry 6 months after surgery may be useful to detect subclinical obstruction.


Development of aquatic life criteria for nitrobenzene in China.
Nitrobenzene is a toxic pollutant and was the main compound involved in the Songhuajiang accident in 2007, one of the largest water pollution accidents in China in the last decade. No aquatic life criteria for nitrobenzene have previously been proposed. In this study, published toxicity data of nitrobenzene to Chinese aquatic species were gathered, and six resident Chinese aquatic organisms were used in toxicity tests to supplement the existing toxicity data for nitrobenzene. Seventeen genuses mean acute values, three genuses mean chronic values to freshwater aquatic animals, and six genus toxicity values to aquatic plants were collected in total. A criterion maximum concentration of 0.018 mg/L and a criterion continuous concentration of 0.001 mg/L were developed based on these data, according to the U.S. Environmental Protection Agency guidelines. These criteria may be useful in the determination of water quality standard of nitrobenzene.


[Cytological diagnosis of Hashimoto's hyperthyroidism].
To assess the value of thyroid fine needle biopsy (FNB) in Hashimoto's hyperthyroidism. FNB for was performed 58 (5 males and 53 females, aged from 13 to 66 years) of 1 031 patients who was performed clinically dignosised as having hyperthyroidism according to clinic symptoms, the results of physical and, laboratory examination, especially the increased the serum circle antibody and lowered sTSH during March 1995 to February 1998. Their smears were classified into 4 types according to the degree of degeneration of adenoidal cell and lymphocyte invasion. Hyperthyroidism was observed in 5 cases, I type in 9, II type in 9, III type in 4, and IV type in 31 (53%). FNB is helpful in diagnosis of hyperthyroidism.


Nitric oxide-based protein modification: formation and site-specificity of protein S-nitrosylation: Could protein S-nitrosylation be the unifying oxidative modification to explain the cellular signaling activity of superoxide and hydrogen peroxide?
Accumulating evidence indicates that reactive oxygen species (ROS) and reactive nitrogen species (RNS) function as signaling molecules in physiological settings by acting as second messengers in response to external stimuli such as growth factors, cytokines and hormones. The nature of the ROS involved in cell signaling as well as the underlying mechanisms by which ROS modify protein function to influence cellular processes have been unfolding over the past decade. ROS and RNS influence various cellular processes by altering the function of critical proteins via reversible oxidation of "reactive cysteine" residues. Protein S-nitrosylation is a mechanism of nitric oxide-based signaling, however, while the presence of NO is sufficient and may be a prerequisite for the formation of cysteine-SNO, we reasoned that if protein-SNO formation is a critical cystein modification for redox driven signal transduction, an increase in intracellular ROS such as H2O2 and O2(-), shown to be independently involved in cell signaling, might both promote the formation of protein-SNO. In this respect, the present study shows that an increase in protein-SNO was detected not only upon an increase in the intracellular level of nitric oxide (NO), but also following exposure to low concentration of exogenous hydrogen peroxide (H2O2) or upon inhibition of the Cu/Zn superoxide dismutase that results in increased intracellular O2(-).


Colony size distribution as a measure of age in cultured human cells. A brief note.
Individual human diploid cells from the same culture differ greatly in the number of doublings they can achieve and consequently the size of colony they can initiate. Computer simulations suggested that the life-span of a culture could be determined from the distribution of colony sizes. Colony size distributions of cultures of a human diploid fibroblast, WI-38, were determined experimentally and found to be a sensitive measure of in vitro age. In particular, there is a highly significant linear correlation between the percentage of colonies of sixteen or more cells and the number of population doublings remaining in the in vitro life-span. Thus, the colony size distribution can be used to predict the number of population doublings remaining in the vitro life-span of a human diploid fibroblast cell culture without knowledge of its prior history.


Management of non-functional pancreatic neuroendocrine tumors by endoscopic ultrasound-guided radiofrequency ablation: Retrospective study in two tertiary centers.
Recently, there has been growing interest in investigating endoscopic ultrasound-guided radiofrequency ablation (EUS-RFA) for the management of small non-functional pancreatic neuroendocrine tumors (nf pNETs). A bicentric retrospective study was performed that included patients with histologically confirmed nf pNETs who were consecutively treated by EUS-RFA between December 2015 and March 2021 at two tertiary referral centers. In 27 patients (mean age 65.0 years, 52% male), EUS-RFA was successfully performed. All patients had sporadic G1 lesions (mean size 14.0 ± 4.6 mm, 7% uncinated process, 22% head, 11% body, 19% body/tail junction, and 41% tail). Overall, 9/27 lesions (33%) were cystic. The mean hospital stay was 3.2 days. Complete treatment response was confirmed in 25/27 patients (93%) on cross-sectional imaging (mean follow-up 15.7 ± 12.2 months, range 2-41 months). Two patients had two EUS-RFA sessions until complete necrosis was observed. Periprocedural acute pancreatitis occurred in 4/27 (14.8%), three of them were treated by endoscopic cystogastrostomy (11.1%). One patient underwent secondary surgery. The histopathology of the resected specimen revealed 3 mm of residual tumor tissue. EUS-RFA seems to be a promising treatment strategy for the management of small nf pNETs with excellent efficacy. Further evidence focusing on long-term survival, safety profile and recurrence is needed.


Modulation of the soleus H-reflex during knee rotations is not consistent with muscle fascicle length changes.
The purpose of this study was to examine whether passively rotating the knee would result in parallel or differential changes to the medial gastrocnemius (MG) and soleus (SOL) H-reflex amplitudes. Since passive knee rotation alters the muscle length of the MG, but not the SOL, it was hypothesized that the MG H-reflex would reflect the lengthening or shortening actions that occur during knee rotation, whereas the SOL H-reflex would remain unaltered. MG and SOL Hoffman reflexes (H-reflexes) were evoked with the knee joint held static at 10° or as the joint was passively flexed or extended past 10°. Ultrasound recordings were used to confirm whether the knee rotations altered MG but not SOL muscle fascicle lengths. In contrast to our hypothesis, results indicated that the MG and SOL H-reflexes were similarly affected during knee rotations, with both MG and SOL H(max):M(max) smaller during the knee extension than the knee flexion (33-43% reduction) and static (22-28% reduction) conditions. Parallel changes to the MG and SOL H-reflexes occurred despite a differential effect of knee rotation on muscle fascicle lengths. Whereas, MG muscle fascicles lengthened and shortened during knee extension and flexion, respectively, SOL fascicles length remained unchanged. Given the strong neural coupling between the MG and SOL motoneuron pools, the results highlight the difficulty in isolating specific variables (e.g., muscle length) when determining the modulatory influences on the triceps surae H-reflex amplitude.


The centenary of the sinking of the Mediterranean Fleet flagship, HMS Victoria. What was the role of Malta fever?
This article commemorates the centenary of the tragic loss of the battleship HMS Victoria and 358 of her crew, together with the Commander-in-Chief of the Mediterranean Fleet, Vice-Admiral Sir George Tryon, as a result of an enigmatic order by Tryon himself. It also investigates the medical aspects and explores the contention that Tryon was suffering from Malta or Mediterranean fever (Brucellosis).


Second-Hit Somatic Mutations in Mevalonate Pathway Genes Underlie Porokeratosis.
Familial and sporadic porokeratosis are associated with germline heterozygous mutations in mevalonate pathway genes. Kubo et al. show that each skin lesion of disseminated superficial actinic porokeratosis originates from a postnatal keratinocyte clone with a different second-hit genetic event in the wild-type allele of the corresponding gene. They also confirm that linear porokeratosis derives from a single prenatal clone of keratinocytes with a second-hit genetic event.


The relation between polarity sensitivity and neural degeneration in a computational model of cochlear implant stimulation.
The main aim of this computational modelling study was to test the validity of the hypothesis that sensitivity to the polarity of cochlear implant stimulation can be interpreted as a measure of neural health. For this purpose, the effects of stimulus polarity on neural excitation patterns were investigated in a volume conduction model of the implanted human cochlea, which was coupled with a deterministic active nerve fibre model based on characteristics of human auditory neurons. The nerve fibres were modelled in three stages of neural degeneration: intact, with shortened peripheral terminal nodes and with complete loss of the peripheral processes. The model simulated neural responses to monophasic, biphasic, triphasic and pseudomonophasic pulses of both polarities. Polarity sensitivity was quantified as the so-called polarity effect (PE), which is defined as the dB difference between cathodic and anodic thresholds. Results showed that anodic pulses mostly excited the auditory neurons in their central axons, while cathodic stimuli generally excited neurons in their peripheral processes or near their cell bodies. As a consequence, cathodic thresholds were more affected by neural degeneration than anodic thresholds. Neural degeneration did not have a consistent effect on the modelled PE values, though there were notable effects of electrode contact insertion angle and distance from the modiolus. Furthermore, determining PE values using charge-balanced multiphasic pulses as approximations of monophasic stimuli produced different results than those obtained with true monophasic pulses, at a degree that depended on the specific pulse shape; in general, pulses with lower secondary phase amplitudes showed polarity sensitivities closer to those obtained with true monophasic pulses. The main conclusion of this study is that polarity sensitivity is not a reliable indicator of neural health; neural degeneration affects simulated polarity sensitivity, but its effect is not consistently related to the degree of degeneration. Polarity sensitivity is not simply a product of the state of the neurons, but also depends on spatial factors.


A preganglionic autonomic nucleus in the dorsal gray commissure of the lumbar spinal cord of the rat.
The distribution in the spinal cord of the rat of preganglionic neurons sending fibers into the hypogastric nerve was determined with the retrograde transport of horseradish peroxidase (HRP). Labeled cells were present in the intermediate gray matter of spinal segments L1-L2. The majority (81%) of HRP-filled cells formed a continuous column along the midline in the dorsal gray commissure. This cell column was termed the dorsal commissural nucleus (DCN). The remainder of HRP-labeled cells were present bilaterally in the middle and lateral regions of the intermediate gray; the majority of the latter cells were located along the lateral border of the intermediate gray. The present finding of a midline preganglionic autonomic cell column in the spinal cord of a mammal is contrary to previous reports, in which sympathetic preganglionic neurons have been localized primarily in the lateral intermediate gray. The DCN may be species-specific and related to the system of short adrenergic neurons present in the pelvis.


Ethoxyresorufin O-deethylase activity in rat brain subcellular fractions.
Non-polar, lipid-soluble drugs or xenobiotics are able to cross the blood-brain barrier. The brain cytochrome P-450-dependent monooxygenases may oxidize these molecules to more polar and somewhat hazardous metabolites responsible for neurotoxicity. In order to characterize the cytochrome P-450 dependent aryl hydrocarbon hydroxylase activity in brain subcellular fractions, we used 7-ethoxyresorufin as a substrate, as its O-deethylation reflects specifically the activity of the cytochrome P-450 isoform which metabolizes and is induced by polycyclic aromatic hydrocarbons. The results reported here show that this enzymatic activity occurs in both microsomal and mitochondrial fractions, and that the induction after 3-methylcholanthrene treatment remains limited, thus preventing the formation of high levels of harmful metabolites.


Microarray-based analysis of gene expression profiles in peripheral blood of patients with acute primary angle closure.
We investigated the expression of molecules in peripheral blood mononuclear cells (PBMCs) and plasma of patients with acute primary angle closure (APAC). Peripheral blood was collected from patients with APAC (n = 10) and age-matched controls (n = 5). The gene transcription profile was analyzed in PBMCs using microarrays and validated by real-time reverse transcription polymerase chain reaction (RT-PCR). The levels of secreted proteins were evaluated in plasma by ELISA. 347 gene transcripts were up-regulated by 2-fold or more, and 696 transcripts down-regulated 2-fold or more in PBMCs from patients compared to controls. The most highly up-regulated gene was thrombospondin-1 (TSP-1, 8.66-fold increase), and the most down-regulated gene was prostaglandin-endoperoxide synthase 2 (PTGS2, 9.09-fold decrease). Real-time RT-PCR assay confirmed the increase of TSP-1 and the decrease of PTGS2 in PBMCs of patients. ELISA revealed that the levels of TSP-1 and active transforming growth factor (TGF)-β1 that is activated by TSP-1 were elevated in plasma of patients, while the level of prostaglandin E2 (PGE2) that is converted by PTGS2 was reduced. The plasma level of TSP-1 was positively correlated with that of active TGF-β1. Our data suggest that the molecular network including TSP-1, TGF-β1, and PGE2 might be involved in the pathogenesis of APAC and PACG.


[Idiopathic mitral subvalvular aneurysm. Report of a case and review of the literature].
An unusual type of annular, subvalvular left ventricular aneurysm is described for the first time in a 64 years-old mexican woman. The pathological characteristics are similar to those cardiac aneurysms of unknown etiology described in African negroes. In this paper, we review the clinical features, diagnostic difficulties and discuss possible etiologic factors related to this condition.


Thrombotic superior vena caval obstruction after repair of tetralogy of Fallot.
A case of tetralogy of Fallot is described, in which thrombotic obstruction of the superior vena cava developed after total correction. Partial thrombolysis was achieved with streptokinase therapy.


The magic spot: a ppGpp binding site on E. coli RNA polymerase responsible for regulation of transcription initiation.
The global regulatory nucleotide ppGpp ("magic spot") regulates transcription from a large subset of Escherichia coli promoters, illustrating how small molecules can control gene expression promoter-specifically by interacting with RNA polymerase (RNAP) without binding to DNA. However, ppGpp's target site on RNAP, and therefore its mechanism of action, has remained unclear. We report here a binding site for ppGpp on E. coli RNAP, identified by crosslinking, protease mapping, and analysis of mutant RNAPs that fail to respond to ppGpp. A strain with a mutant ppGpp binding site displays properties characteristic of cells defective for ppGpp synthesis. The binding site is at an interface of two RNAP subunits, ω and β', and its position suggests an allosteric mechanism of action involving restriction of motion between two mobile RNAP modules. Identification of the binding site allows prediction of bacterial species in which ppGpp exerts its effects by targeting RNAP.


A mouse c-ros genomic clone: identification of a highly conserved 22-amino acid segment in the juxta-membrane domain.
The mouse c-ros protooncogene genomic sequences have been cloned; an analysis of the partial genomic clone revealed a high conservation of the exons encoding the juxta-membrane (JM) and the 5' most protein tyrosine kinase domains. We have identified a segment of 22 amino acids conserved between the human and mouse JM domains; this segment may have a critical role in the function of the c-ros-encoded protein tyrosine kinase receptor.


Inflammatory cytokines enhance procoagulant activity of platelets and endothelial cells through phosphatidylserine exposure in patients with essential hypertension.
The exact mechanism of the prothrombotic state of essential hypertension (EH) patients remains elusive. Our objective was to assess whether phosphatidylserine (PS) exposure on endothelial cells (ECs), platelets, and microparticles (MPs) can account for the hypercoagulability in EH patients. PS exposure on cells and MPs, mainly from platelets and ECs was analyzed with flow cytometry. Procoagulant activity (PCA) was evaluated by purified coagulation complex assays, clotting time, and fibrin turbidity. We found that EH patients exhibited elevated levels of PS+ platelets, serum-cultured ECs, MPs, endothelial-derived MPs and platelet-derived MPs compared to the controls (all P < 0.05). Moreover, platelets and MPs from the patients and their sera-cultured ECs showed markedly enhanced intrinsic/extrinsic FXa, thrombin, and fibrin generation, and greatly shortened coagulation time. This PCA could be blocked approximately 80%, by the addition of lactadherin. Furthermore, we detected elevated levels of IL-8, IL-6, and TNF-α in EH patients could activate platelets/ECs and induce elevated PS exposure on their membranes. Our results suggest that inflammatory cytokines could enhance procoagulant activity of platelets and endothelial cells via their PS exposure in EH patients. As such, a PS blockade may be a viable therapeutic strategy for treating such patients.


Synthesis of β-arabinofuranoside glycolipids, studies of their binding to surfactant protein-A and effect on sliding motilities of M. smegmatis.
Surfactant protein A (SP-A), which is a lung innate immune system component, is known to bind glycolipids present at the cell surface of a mycobacterial pathogen. Lipoarabinomannan (LAM), a component of mycobacterial thick, waxy cell wall, is one of the glycolipid ligands for SP-A. In order to assess binding of synthetic glycolipids with SP-A and the glycosidic linkage preferences for the interaction, β-arabinofuranoside trisaccharide glycolipids constituted with β-(1→2), β-(1→3) and β-(1→2), β-(1→5) linkages relevant to LAM were synthesized through chemical glycosylations. The efficacies of synthetic glycolipids to interact with SP-A were assessed by using the surface plasmon resonance (SPR) technique, from which association-dissociation rate constants and equilibrium binding constants were derived. The equilibrium binding constants of the interaction of two constitutionally varying β-arabinofuranoside glycolipids with SP-A were found to be in the millimolar range. A comparison of the results with few α-anomeric arabinofuranoside glycolipids showed that glycolipids with β-anomeric linkages were having relatively lower equilibrium binding constants than those with α-anomeric linkages in binding to the protein, whereas oligosaccharides alone, without lipidic chains, exhibited higher equilibrium binding constants. Further, the synthetic compounds inhibited the growth of mycobacteria and affected sliding motilities of the bacteria, although to an extent relatively lesser than that of synthetic compounds constituted with α-anomeric linkages.


Expanding the knowledge around antitubercular 5-(2-aminothiazol-4-yl)isoxazole-3-carboxamides: Hit-to-lead optimization and release of a novel antitubercular chemotype via scaffold derivatization.
Tuberculosis is one of the deadliest infectious diseases in the world, and the increased number of multidrug-resistant and extensively drug-resistant strains is a reason for concern. We have previously reported a series of substituted 5-(2-aminothiazol-4-yl)isoxazole-3-carboxamides with growth inhibitory activity against Mycobacterium tuberculosis strains and low propensity to be substrate of efflux pumps. Encouraged by these preliminary results, we have undertaken a medicinal chemistry campaign to determine the metabolic fate of these compounds and to delineate a reliable body of Structure-Activity Relationships. Keeping intact the (thiazol-4-yl)isoxazole-3-carboxamide core, as it is deemed to be the pharmacophore of the molecule, we have extensively explored the structural modifications able to confer good activity and avoid rapid clearance. Also, a small set of analogues based on isostere manipulation of the 2-aminothiazole were prepared and tested, with the aim to disclose novel antitubercular chemotypes. These studies, combined, were instrumental in designing improved compounds such as 42g and 42l, escaping metabolic degradation by human liver microsomes and, at the same time, maintaining good antitubercular activity against both drug-susceptible and drug-resistant strains.


AlgPred: prediction of allergenic proteins and mapping of IgE epitopes.
In this study a systematic attempt has been made to integrate various approaches in order to predict allergenic proteins with high accuracy. The dataset used for testing and training consists of 578 allergens and 700 non-allergens obtained from A. K. Bjorklund, D. Soeria-Atmadja, A. Zorzet, U. Hammerling and M. G. Gustafsson (2005) Bioinformatics, 21, 39-50. First, we developed methods based on support vector machine using amino acid and dipeptide composition and achieved an accuracy of 85.02 and 84.00%, respectively. Second, a motif-based method has been developed using MEME/MAST software that achieved sensitivity of 93.94 with 33.34% specificity. Third, a database of known IgE epitopes was searched and this predicted allergenic proteins with 17.47% sensitivity at specificity of 98.14%. Fourth, we predicted allergenic proteins by performing BLAST search against allergen representative peptides. Finally hybrid approaches have been developed, which combine two or more than two approaches. The performance of all these algorithms has been evaluated on an independent dataset of 323 allergens and on 101 725 non-allergens obtained from Swiss-Prot. A web server AlgPred has been developed for the predicting allergenic proteins and for mapping IgE epitopes on allergenic proteins (http://www.imtech.res.in/raghava/algpred/). AlgPred is available at www.imtech.res.in/raghava/algpred/.


Solving the heart mechanics equations with Newton and quasi Newton methods--a comparison.
The non-linear elasticity equations of heart mechanics are solved while emulating the effects of a propagating activation wave. The dynamics of a 1 cm(3) slab of active cardiac tissue was simulated as the electrical wave traversed the muscular heart wall transmurally. The regular Newton (Newton-Raphson) method was compared to two modified Newton approaches, and also to a third approach that delayed update only of some selected Jacobian elements. In addition, the impact of changing the time step (0.01, 0.1 and 1 ms) and the relative non-linear convergence tolerance (10(-4), 10(-3) and 10(-2)) was investigated. Updating the Jacobian only when slow convergence occurred was by far the most efficient approach, giving time savings of 83-96%. For each of the four methods, CPU times were reduced by 48-90% when the time step was increased by a factor 10. Increasing the convergence tolerance by the same factor gave time savings of 3-71%. Different combinations of activation wave speed, stress rate and bulk modulus revealed that the fastest method became relatively even faster as stress rate and bulk modulus was decreased, while the activation speed had negligible influence in this respect.


Aortico-right atrial tunnel in an adult patient.
We describe an asymptomatic 27-year-old man who presented a continuous murmur, and was found to have a tunnel between the aorta and the right atrium. Successful surgical treatment was performed. We emphasize the rarity of this condition, and discuss its major features.


Quantification of the electron donating capacity and UV absorbance of dissolved organic matter during ozonation of secondary wastewater effluent by an assay and an automated analyzer.
Ozonation of secondary wastewater treatment plant effluent for the abatement of organic micropollutants requires an accurate process control, which can be based on monitoring ozone-induced changes in dissolved organic matter (DOM). This study presents a novel automated analytical system for monitoring changes in the electron donating capacity (EDC) and UV absorbance of DOM during ozonation. In a first step, a quantitative photometric EDC assay was developed based on electron-transfer reactions from phenolic moieties in DOM to an added chemical oxidant, the radical cation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS·+). The assay is highly sensitive (limit of quantification ∼0.5 mgDOC·L-1) and EDC values of model DOM isolates determined by this assay were in good agreement with values determined previously by mediated electrochemical oxidation (slope = 1.01 ± 0.07, R2 = 0.98). In a second step, the photometric EDC measurement method was transferred onto an automated fluidic system coupled to a photometer (EDC analyzer). The EDC analyzer was then used to monitor changes in EDC and UV absorbance of secondary wastewater effluent treated with ozone. While both parameters exhibited a dose-dependent decrease, a more pronounced decrease in EDC as compared to UV absorbance was observed at specific ozone doses up to 0.4 mgO3·gDOC-1. The concentration of 17α-ethinylestradiol, a phenolic micropollutant with a high ozone reactivity, decreased proportionally to the EDC decrease. In contrast, abatement of less ozone-reactive micropollutants and bromate formation started only after a pronounced initial decrease in EDC. The on-line EDC analyzer presented herein will enable a comprehensive assessment of the combination of EDC and UV absorbance as control parameters for full-scale ozonation.


Comparison of the effects of sensorimotor training programs on pain, electromyography and kinematics in patients with scapular downward rotation syndrome.
Comparison of the effects of six weeks sensorimotor and sensorimotor with passive interventions programs on pain, electromyography (EMG) and kinematics in patients with scapular downward rotation syndrome (SDRS). Randomized Controlled Trial. Institutional practice. 140 active subjects with unilateral SDRS were randomized to three groups. Group one received sensorimotor (n = 46), group two received sensorimotor with passive interventions (n = 48), and group three received active self-exercise as a control group (n = 46). Pain, EMG of the levator scapula (LS), upper trapezius (UT), lower trapezius (LT), and serratus anterior (SA), as well as kinematics were measured at the baseline and after the interventions. Primary outcome: Pain - Secondary outcomes: EMG and kinematics. There were significant between-group differences between intervention groups one and two in pain, LS and SA onset activation favoring group two and LS muscle activity favoring group one. There were significant within-group changes in almost all dependent variables except LT muscle onset activation in both groups one and two. The addition of passive interventions on the scapula and neck may be superior to conservative training alone on the scapula and neck for improving neck pain, EMG and kinematics in participants with SDRS.


Zika virus infects human blood mononuclear cells.
Zika virus (ZIKV) infection gained public health concern after the 2015 outbreak in Brazil, when microcephaly rates increased in babies born from infected mothers. It was demonstrated that ZIKV causes a congenital Zika virus syndrome, including various alterations in the development of the central nervous system. Although the infection of cells from the nervous system has been well documented, less is known in respect of ZIKV ability to infect immune cells. Herein, we investigated if peripheral blood mononuclear cells (PBMCs), freshly-isolated from healthy donors, could be infected by ZIKV. PBMCs from healthy donors were isolated and cultured in medium with ZIKV strain Rio-U1 (MOI = 0.1). Infection was analyzed by RT-qPCR and flow cytometry. We detected the ZIKV RNA in PBMCs from all donors by RT-qPCR analysis. The detection of viral antigens by flow cytometry revealed that PBMC from more than 50% the donors were infected by ZIKV, with CD3+CD4+ T cells, CD3-CD19+ B cells and CD3+CD8+ T cells being, respectively, the most frequently infected subpopulations, followed by CD14+ monocytes. Additionally, we observed high variability in PBMC infection rates among different donors, either by numbers or type infected cells. These findings raise the hypothesis that PBMCs can act as a reservoir of the virus, which may facilitate viral dissemination to different organs, including immune-privileged sites.


Virulence traits and antibiotic resistance among enterococci isolated from dogs with periodontal disease.
Periodontal disease - PD - is one of the most widespread diseases in dogs, but the role of this odontogenic infection in the dissemination of pathogenic bacteria present in the oral mucosa to other animals or pet owners is understudied. Trying to unveil the putative pathogenicity of enterococci present in the gums of dogs diagnosed with PD, thirty-two animals were investigated during routine visits to a private veterinary clinic. Seventy-one enterococci were recovered and characterized regarding species, genomic variability, virulence traits, antimicrobial resistance and biofilm-forming ability. Isolates were mainly identified as Enterococcus faecalis, with the large majority (95%) being able to produce biofilm. Regarding antibiotic resistance, all dog-enterococci were susceptible to ampicillin, amoxicillin/clavulanate, gentamicin-120, imipenem and vancomycin; while distinct levels of resistance were observed for chloramphenicol (10%), erythromycin (20%), streptomycin-300 (35%) and tetracycline (95%). For virulence traits incidence levels of 35% were observed for β-hemolysis and 25% for cylA, 25% for gelatinase and 35% for gelE; 85% harbor efaAfs and ebpABC; while ace, agg and esp are present respectively in 50, 30 and 10% of the dog-enterococci; efaAfm and acm were detected in all the Enterococcus faecium. Overall, the widespread prevalence of PD in dogs, associated with the close contact between companion animals, other animals and humans, may act as source for the dissemination of opportunistic pathogenic bacteria. Hence, aforementioned data on virulence and resistance features, emphasizes the need for active surveillance measures, such as the diagnose of PD in companion animals during routine visits to the veterinary clinic.


Efficacy and tolerance of spironolactone in essential hypertension.
The long-term efficacy and tolerance of spironolactone in essential hypertension was evaluated among 20,812 patients referred to the Broussais and St. Joseph systemic hypertension clinics between 1976 and 1985 by using information prospectively collected in the computerized ARTEMIS data bank. In 182 patients (51 men, 131 women) treated with spironolactone alone during a mean follow-up period of 23 months, a mean dose of 96.5 mg decreased systolic and diastolic blood pressure (BP) by 18 and 10 mm Hg, respectively, below pretherapeutic levels. The BP decrease was greater with doses of 75 to 100 mg (12.4% and 12.2%) than with doses of 25 to 50 mg (5.3 and 6.5%, p less than 0.001), but no additional decrease was found with doses above 150 mg. Plasma creatinine level increased modestly (8.3 mumol/liters), as did plasma potassium level (0.6 mmol/liters) (both p less than 0.001); uric acid level increased, but not significantly (10.5 mumol/liter). Fasting blood glucose and total cholesterol levels did not change, triglyceride levels increased slightly (0.1 mmol/liter, p less than 0.05). These changes were similar in both sexes and were not influenced by length of follow-up. Among the 699 men prescribed spironolactone alone or in association with another antihypertensive treatment, 91 cases of gynecomastia developed (13%). Gynecomastia was reversible and dose-related; at doses of 50 mg or less the incidence was 6.9%, but 52.2% for doses of 150 mg or higher. Despite limitations inherent in the interpretation of data banks, it is concluded that spironolactone administered in daily practice reduced BP without inducing adverse metabolic adverse effects and that in patients with essential hypertension, doses should be kept below 100 mg.


Activity and reproductive state in the hamster: independent control by social stimuli and a circadian pacemaker.
Entrainment of circadian rhythms by social communication between male and female Syrian hamsters (Mesocricetus auratus) was tested by recording the wheel-running activity of pairs kept in the same cage but separated by a wire mesh barrier. Before pairing, males and females were synchronized to light/dark cycles that were 180 degrees out of phase, and at the time of pairing the hamsters were placed into constant darkness (DD). The activity rhythms of males and females housed in a cage alone (isolated) were also recorded. The freerunning periods of paired and isolated hamsters were not different over six weeks in DD, and no phase-shifts of the paired animals' rhythms were seen, indicating that the close proximity of a hamster of the opposite sex had no effect on the timing of the other's activity/rest rhythm. This was not due to a lack of communication between the paired males and females. Males showed four-day cycles in the amount and distribution of activity which corresponded to the estrous cycle of the female, and regression of the reproductive system which occurred in the isolated hamsters was delayed in both the paired males and females. Despite the fact that locomotor activity and reproduction are each regulated in part by a circadian pacemaker, social stimuli can affect both of these without influencing the circadian pacemaker that underlies the activity/rest rhythm.


Are there any differences between different testing sites? A cross-sectional study of a Norwegian low-threshold HIV testing service for men who have sex with men.
To describe a Norwegian low-threshold HIV testing service targeting men who have sex with men (MSM). After the HIV testing consultation, all users of the HIV testing service were invited to answer the study questionnaire. The study setting included the sites where testing was performed, that is, the testing service's office in Oslo, cruising areas, bars/clubs and in hotels in other Norwegian cities. MSM users of the testing service. Data were collected on demographics, HIV testing and sexual behaviour as well as the participant's motivations for choosing to take an HIV test at this low-threshold HIV testing service. The data are stratified by testing site. 1577 HIV testing consultations were performed, the study sample consisted of 732 MSM users. 11 tested positive for HIV. 21.7% had a non-western background, 27.1% reported having a non-gay sexual orientation. 21.9% had 10 or more male sexual partners during the last year, 27.9% reported also having had a female sexual partner. 56.4% reported having practised unprotected anal intercourse during the last 6 months. 20.1% had never tested for HIV before. Most of these user characteristics varied by testing sites. The Norwegian low-threshold testing service recruits target groups that are otherwise hard to reach with HIV testing. This may indicate that the testing service contributes to increase HIV testing rates among MSM in Norway.


Association between leukaemia inhibitory factor gene polymorphism and pregnancy outcomes after assisted reproduction techniques.
Certain gene polymorphisms are associated with implantation failure and pregnancy loss. Studies of leukaemia inhibitory factor (LIF) gene polymorphisms are scarce. The LIF single nucleotide polymorphism (SNP) thymine (T)/guanine (G) (rs929271) was studied in women to determine whether an association existed with pregnancy outcomes after intracytoplasmic sperm injection (ICSI); 411 women who underwent ICSI were recruited. DNA was extracted from the peripheral blood, and the LIF gene SNP T/G (rs929271) was genotyped using real-time polymerase chain reaction. Participants were divided into three groups according to their LIF genotype: T/T (n = 168), T/G (n = 202) and G/G (n = 41). All IVF and ICSI procedures were carried out under the same clinical and laboratory conditions. The ICSI cumulative results (from fresh plus frozen cycles) of each genotype group were analysed. The G/G genotype in women was associated with a higher implantation rate (T/T: 15.9%, T/G: 16.2%, G/G: 27.0%; P < 0.05), ongoing pregnancy rate/patient (T/T: 31.5%, T/G: 36.1%, G/G: 53.7%; P < 0.05) and ongoing pregnancy rate/transfer (T/T: 18.5%, T/G: 20.2%, G/G: 36.7%; P < 0.05). LIF SNP T/G (rs929271) seems to be a susceptibility biomarker capable of predicting implantation efficiency and pregnancy outcomes.


Kinetic study on the production and degradation of leupeptin in Streptomyces exfoliatus SMF13.
Medium composition and cultivation conditions were constructed for the optimum production of leupeptin by Streptomyces exfoliatus SMF13. The production of leupeptin was related to mycelial growth, being optimum in the cultivation with glucose-excess, phosphate-limited, and casamino acids medium. However, leupeptin-inactivating enzyme (LIE) was produced in the cultivation with glucose-limited, phosphate-excess, and Na-caseinate medium where mycelium degradation was accompanied. LIE was one of the most important factors in optimizing the leupeptin productivity. Kinetic parameters calculated from batch and chemostat cultivations revealed that qlpt was closely related to qs and mu, but qLIE was increased after mu declined to near zero, and followed by kd. Optimum production process of leupeptin was determined with phosphate-limited continuous cultivation, which did not permit LIE production. The maximum productivity (0.24 g l-1 h-1) and production yield (1.64 g leupeptin per g glucose) of phosphate-limited chemostat cultivation were 2.4- and 4-times larger than those of batch cultivation, respectively. This is the first cultivation kinetic analysis for leupeptin production and its inactivation by LIE in relation to mycelium differentiation.


Leucocyte chemotaxis: Examination of mitogen-activated protein kinase and phosphoinositide 3-kinase activation by Monocyte Chemoattractant Proteins-1, -2, -3 and -4.
Monocyte Chemoattractant Proteins (MCPs) form a distinct, structurally-related subclass of CC chemokines. They are major chemoattractants for monocytes and T lymphocytes. The MCPs bind to specific G-protein-coupled receptors, initiating a signal cascade within the cell. Though the signal transduction pathways involved in MCP-1-mediated chemotaxis have been studied, the signalling pathways through which MCP-2, -3 and -4 trigger cell migration are not established. In this study, we examined the mitogen-activated protein kinase (MAPK) activation elicited by the MCPs (1-4) and its specific role in chemotaxis. Within 2 min, the MCPs (1-4) elicited a rapid and transient activation of MAPK in peripheral blood mononuclear cells and in HEK-293 cells expressing CCR2b. U0126, an inhibitor of MAPK-kinase (MEK) activation, not only prevented extracellular signal-regulated kinase 1/2 (ERK1/2) activation but also significantly inhibited the MCP-mediated chemotaxis. PI3K inhibitors Wortmannin and LY294002 also partially inhibited the MCP-induced chemotaxis. However, these compounds did not significantly inhibit ERK1/2 activation. As PI3K inhibitors partially inhibit the MCP-mediated chemotaxis but do not significantly effect ERK1/2 activation, these data suggest that co-ordinated action of distinct signal pathways is required to produce chemokine-mediated chemotaxis.


Rapid isolation of viral integration site reveals frequent integration of HTLV-1 into expressed loci.
Although there is tight association of the human T-cell leukemia virus type-1 (HTLV-1) with adult T-cell leukemia/lymphoma (ATLL), it has remained unresolved whether the HTLV-1 integration into the host genome has any role in the development of this disease. We isolated a total of 58 HTLV-1 integration sites using newly developed, adaptor-ligated PCR from 33 ATLL patients and five ATLL cell lines. We compared our data as well as the previously reported ones with the complete human genomic sequence for the location of its placement, structure, and expression of genes nearby the integration site. The chromosomal target for integration was selected at random, but the integration favorably occurred within the transcription units; more than 59.5% of total integration was observed within the transcriptional unit. All inserted genes by HTLV-1 integration were expressed in normal T cells. Upregulation of genes due to viral integration was found in two out of nine ATLL cases; about 4.4- and 102-fold elevated ankyrin-1 ( ANK-1) and gephyrin ( GPHN) gene expressions were observed, respectively. These data suggest that the preferential integration of HTLV-1 into an expressed locus occasionally causes deregulation of corresponding gene, which may lead to leukemogenesis of a fraction of ATLL.


Vestibular Evoked Myogenic Potential (VEMP) Test-retest Reliability in Children.
Vestibular evoked myogenic potentials (VEMPs) are short-latency muscle potentials measured from the neck (cervical VEMP; cVEMP) or under the eyes (ocular VEMP; oVEMP), which provide information regarding function of the saccule and utricle, respectively. VEMPs are reliable when performed in adults; however, reliability of VEMPs in children is unknown. Therefore, the purpose of the study was to determine the test-retest reliability of c- and oVEMP testing in normal control children. Prospective. Hospital. Ten adults, 14 adolescent children and 13 young children with normal hearing. c- and oVEMP testing were completed across two test sessions in response to air-conduction 500 Hz tone-burst and impulse hammer stimuli. Additionally, oVEMP was completed using eyes-open and eyes-closed conditions. Intraclass correlation coefficients were calculated to determine the reliability of c- and oVEMP outcomes. When using air-conduction stimuli, c- and oVEMP amplitudes are reliable across test sessions in normal control children and adults. With impulse hammer stimuli, cVEMP amplitudes showed high reliability; however, oVEMP amplitudes showed low reliability in both eyes-open and eyes-closed conditions. Comparison between eyes-open and eyes-closed oVEMP conditions revealed shorter latencies and higher peak-to-peak amplitudes in the eyes-open condition. In this small cohort of normal control children, cVEMPs are reliable using air-conduction and impulse hammer stimuli and oVEMPs are reliable using air-conduction stimuli in the eyes-open condition. oVEMP in eyes-closed conditions were less reliable compared with eyes-open conditions and resulted in a large number of absent responses.


Profiling cell envelope-antibiotic interactions reveals vulnerabilities to β-lactams in a multidrug-resistant bacterium.
The cell envelope of Gram-negative bacteria belonging to the Burkholderia cepacia complex (Bcc) presents unique restrictions to antibiotic penetration. As a consequence, Bcc species are notorious for causing recalcitrant multidrug-resistant infections in immunocompromised individuals. Here, we present the results of a genome-wide screen for cell envelope-associated resistance and susceptibility determinants in a Burkholderia cenocepacia clinical isolate. For this purpose, we construct a high-density, randomly-barcoded transposon mutant library and expose it to 19 cell envelope-targeting antibiotics. By quantifying relative mutant fitness with BarSeq, followed by validation with CRISPR-interference, we profile over a hundred functional associations and identify mediators of antibiotic susceptibility in the Bcc cell envelope. We reveal connections between β-lactam susceptibility, peptidoglycan synthesis, and blockages in undecaprenyl phosphate metabolism. The synergy of the β-lactam/β-lactamase inhibitor combination ceftazidime/avibactam is primarily mediated by inhibition of the PenB carbapenemase. In comparison with ceftazidime, avibactam more strongly potentiates the activity of aztreonam and meropenem in a panel of Bcc clinical isolates. Finally, we characterize in Bcc the iron and receptor-dependent activity of the siderophore-cephalosporin antibiotic, cefiderocol. Our work has implications for antibiotic target prioritization, and for using additional combinations of β-lactam/β-lactamase inhibitors that can extend the utility of current antibacterial therapies.


Change in subfoveal choroidal thickness in diabetes and in various grades of diabetic retinopathy.
To evaluate subfoveal choroidal thickness (SFCT) change in diabetes and in various grades of diabetic retinopathy (DR) in comparison to age-matched healthy subjects. This prospective observational study included 100 eyes of diabetic patients without DR (group D), 100 eyes with DR (group R), and 100 eyes of healthy subjects (group N). The assessment included demographics, duration of diabetes, comprehensive ocular examination, fundus photography with/without fundus fluorescein angiography, spectral domain optical coherence tomography with enhanced depth imaging to assess SFCT. The SFCT was comparable between groups N (310.65 ± 37.34 µm) and D (308.48 ± 30.06 µm; P = 0.60), but was significantly lower in R (296.52 ± 21.41 µm; P < 0.01). The SFCT was significantly lower in proliferative DR (n = 36; SFCT = 284.56 ± 21.09 µm) as compared to non-proliferative DR (n = 64; SFCT = 303.25 ± 18.59 µm; P < 0.001). The SFCT had moderately negative correlation with severity of DR (R = - 0.50; P < 0.01). The difference in SFCT when compared with normal subjects was significant only in severe/very severe non-proliferative DR (294.47 ± 15.65 µm; P < 0.01) and in proliferative DR (284.56 ± 21.09 µm; P < 0.01). There was a negative correlation of SFCT with the duration of diabetes (R = - 0.41; P < 0.01). SFCT decreases with increasing duration of diabetes. The decrease is significant after the onset of severe DR, and is proportionate to the severity of DR.


Effect of restoration volume on stresses in a mandibular molar: a finite element study.
There can be significant disagreement among dentists when planning treatment for a tooth with a failing medium-to-large--sized restoration. The clinician must determine whether the restoration should be replaced or treated with a crown, which covers and protects the remaining weakened tooth structure during function. The purpose of this study was to evaluate the stresses generated in different sized amalgam restorations via a computational modeling approach and reveal whether a predictable pattern emerges. A computer tomography scan was performed of an extracted mandibular first molar, and the resulting images were imported into a medical imaging software package for tissue segmentation. The software was used to separate the enamel, dentin, and pulp cavity through density thresholding and surface rendering. These tissue structures then were imported into 3-dimensional computer-aided design software in which material properties appropriate to the tissues in the model were assigned. A static finite element analysis was conducted to investigate the stresses that result from normal occlusal forces. Five models were analyzed, 1 with no restoration and 4 with increasingly larger restoration volume proportions: a normal-sized tooth, a small-sized restoration, 2 medium-sized restorations, and 1 large restoration as determined from bitewing radiographs and occlusal surface digital photographs. The resulting von Mises stresses for dentin-enamel of the loaded portion of the tooth grew progressively greater as the size of the restoration increased. The average stress in the normal, unrestored tooth was 4.13 MPa, whereas the smallest restoration size increased this stress to 5.52 MPa. The largest restoration had a dentin-enamel stress of 6.47 MPa. A linear correlation existed between restoration size and dentin-enamel stress, with an R(2) of 0.97. A larger restoration volume proportion resulted in higher dentin-enamel stresses under static loading. A comparison of the von Mises stresses to the yield strengths of the materials revealed a relationship between a tooth's restoration volume proportion and the potential for failure, although factors other than restoration volume proportion may also impact the stresses generated in moderate-sized restorations.


Thalamic projections to the feline motor cortex studied with horseradish peroxidase.
Cells in the thalamus projecting to the distal forelimb regions of the motor cortex, area 3a, and the sensory cortex were identified with horseradish peroxidase (HRP). These cortical areas were defined by mapping evoked potentials from superficial and deep radial nerve stimulation. Following injections into the motor cortex, the labelled cells were distributed throughout a large region (greater than 2 mm wide) in the ventral portion of the ventral lateral nucleus (VL). The border area between VL and the ventral posterolateral nucleus (VPL) also contained labelled cells. The region containing labelled cells following injections into area 3a was in the rostral, dorsal VPL, and overlapped with the region containing cells labelled by injections into the motor cortex. Cells labelled by injections into the sensory cortex were near the center of VPL distinctly separated from those labelled after injections into either area 3a or the motor cortex.


Gossypiboma mimicking posterior urethral stricture.
Foreign bodies in the urogenital tract are not uncommon. Hairpins, glass rods, umbilical tapes, ball point pen are described in lower urogenital tract. Retained gauze piece (gossypiboma) in posterior urethra may cause diagnostic dilemma. Symptoms and investigations may mimic stricture of posterior urethra. Two cases of retained gauze pieces in the urethra are described here. The micturating cystourethrogram was suggestive of posterior urethral stricture. Two cases described here had retained gauze piece as a cause of filling defect and abnormal appearance in the micturating cystourethrogram. Gossypiboma may be a possibility where posterior urethral stricture are seen after previous surgery in paediatric age group. In the setting of previous urogenital surgery gossypiboma should be kept in the differential diagnosis where posterior urethral stricture are seen in the paediatric age group.


Protein modifications involved in neurotransmitter and gasotransmitter signaling.
Covalent modifications of intracellular proteins, such as phosphorylation, are generally thought to occur as secondary or tertiary responses to neurotransmitters, following the intermediation of membrane receptors and second messengers such as cyclic AMP. By contrast, the gasotransmitter nitric oxide directly S-nitrosylates cysteine residues in diverse intracellular proteins. Recently, hydrogen sulfide has been acknowledged as a gasotransmitter, which analogously sulfhydrates cysteine residues in proteins. Cysteine residues are also modified by palmitoylation in response to neurotransmitter signaling, possibly in reciprocity with S-nitrosylation. Neurotransmission also elicits sumoylation and acetylation of lysine residues within diverse proteins. This review addresses how these recently appreciated protein modifications impact our thinking about ways in which neurotransmission regulates intracellular protein disposition.


A computed tomography study of Alzheimer's disease.
Computed tomography (CT) was used to study cerebral atrophy in 18 patients with clinically diagnosed Alzheimer's disease of presenile type and in 14 healthy age-matched subjects as controls. Using the computerized planimetric method, Subarachnoid Space Volume Index and Ventricle Volume Index were calculated as the measure of cortical atrophy and ventricular dilatation respectively. From the results the following conclusions were drawn: 1. The cerebral atrophy in Alzheimer patients could be attributable to the disease processes rather than to physiological aging of the brain. 2. The degree of atrophy increases in parallel with the progress of the clinical stage, and the cortical atrophy is already apparent at an early stage, whereas the ventricular dilatation becomes pronounced at later stages. 3. CT could be one of the most useful clinical tests available for the diagnosis of Alzheimer's disease.


The role of iron in mitochondrial function.
Iron is an essential element for life, as it is a cofactor for enzymes involved in many metabolic processes, but it can also be harmful, since its excess is thought to enhance the production of reactive oxygen species and induce oxidative damage. Iron is transformed into its biologically available form in the mitochondrion by the iron-sulfur (Fe/S) cluster and heme synthesis pathways. During the past decade, substantial progress has been made in the elucidation of iron-linked mechanisms that occur in the mitochondrion, demonstrating the crucial role played by this organelle in maintaining cellular iron homeostasis. This review summarizes current knowledge of the mechanisms underlying iron trafficking in mitochondria and how it is handled inside the organelle. Relevant updates with regard to the Fe/S cluster and heme biosynthetic pathways, as well as the relationship between mitochondrial iron homeostasis impairment and related diseases, are also discussed.


The impact of e-cigarette exposure on different organ systems: A review of recent evidence and future perspectives.
The use of electronic cigarettes (e-cigs) is rapidly increasing worldwide and is promoted as a smoking cessation tool. The impact of traditional cigs on human health has been well-defined in both animal and human studies. In contrast, little is known about the adverse effects of e-cigs exposure on human health. This review summarizes the impact of e-cigs exposure on different organ systems based on the rapidly expanding recent evidence from experimental and human studies. A number of growing studies have shown the adverse effects of e-cigs exposure on various organ systems. The summarized data in this review indicate that while e-cigs use causes less adverse effects on different organs compared to traditional cigs, its long-term exposure may lead to serious health effects. Data on short-term organ effects are limited and there is no sufficient evidence on long-term organ effects. Moreover, the adverse effects of secondhand and third hand e-cigs vapour exposure have not been thoroughly investigated in previous studies. Although some studies demonstrated e-cigs used as a smoking cessation tool, there is a lack of strong evidence to support it. While some researchers suggested e-cigs as a safer alternative to tobacco smoking, their long-term exposure health effects remain largely unknown. Therefore, more epidemiological and prospective studies including mechanistic studies are needed to address the potential adverse health effects of e-cigs to draw a firm conclusion about their safe use. A wide variation in e-cigs products and the lack of standardized testing methods are the major barriers to evaluating the existing data. Specific regulatory guidelines for both e-cigs components and the manufacturing process may be effective to protect consumer health.


Functional differentiation in the anterior gills of the aquatic air-breathing fish, Trichogaster leeri.
The fish gill is a multifunctional organ responsible for gas exchange and ionic regulation. It is hypothesized that both morphological and functional differentiation can be found in the gills of the aquatic air-breathing fish, Trichogaster leeri. To test this, we used the air-breathing fish, Trichogaster leeri, to investigate various morphological/functional parameters. First, we evaluated the importance of performing the aquatic surface respiration behavior in T. leeri. A reduced survival rate was observed when fish were kept in the restrained cages in hypoxic conditions. On the gross anatomy of gills, we found evidence of both morphological and functional modification in the first and the second gills and are responsible for ionic regulation. There were large-bore arterioarterial shunts in the fourth gill arch. It is specialized for the transport of oxygenated blood and is less responsive to environmental stress. In addition, the anterior and the posterior gills differed in the Na(+), K(+)-ATPase activity upon ionic stresses. That is, only the Na(+), K(+)-ATPase activity of the anterior two gills was up-regulated significantly in the deionized water. Lastly, we found that the number of mitochondria-rich cells in the first and the second gills increased following ionic stress and no difference was found in the third and the fourth gills following such an exposure. These results supported the hypothesis that there are morphological and functional differences between anterior and posterior gill arches within the air-breathing Trichogaster leeri. In contrast, no significant difference was found among gills in gross anatomy, filament density and Na(+), K(+)-ATPase activity in the non-air-breather, Barbodes schwanenfeldi.


Children's immigrant generational status and delinquency: The mediating effects of friendship networks.
A mounting body of empirical studies demonstrates that first-generation immigrant children have a lower level of delinquency and crime but second and third-plus generations report a precipitous increase in these behaviors. Adopting a social network approach, we analyzed the behavioral and structural characteristics of children's friendship networks across the first, second, and third-plus immigrant generations, and investigated the mediating role of these friendship traits in explaining generational disparity. Our results reveal that children's friendship networks differ in structural (e.g., popularity) and behavioral features (e.g., network deviance) across immigrant generations. These friendship features, particularly network peer deviance, the percentage of second-generation friends, and children's popularity mediate the association between immigrant generational status and children's delinquency. Extending previous research, our study highlights the importance of applying the social network approach to understand delinquency disparity across immigrant generations and suggests that the composition of friendship networks play an important role in immigrant children's delinquency involvement.


Nucleotide sequence and translation of satellite tobacco mosaic virus RNA.
Satellite tobacco mosaic virus (STMV) is a plant virus with a 17-nm icosahedral particle encapsidating a 0.3 X 10(6) Mr ssRNA genome that depends on tobamoviruses for its replication. The complete nucleotide sequence of STMV RNA deduced in the experiments described here was 1059 nucleotides in length. The efficiency of labeling viral RNA with [gamma-32P]ATP using T4 polynucleotide kinase was not affected by treatment with tobacco acid pyrophosphatase and/or bacterial alkaline phosphatase, indicating that the majority of the 5' termini of encapsidated STMV RNAs were not phosphorylated. The 240 3'-terminal nucleotides of STMV RNA and either tobacco mosaic virus (TMV) U1 RNA or TMV U2/U5 RNA had greater than 65% overall sequence similarity, with two nearly identical regions of 40 and 50 bases, respectively. There were no other regions of sequence relatedness to TMV RNA. The 19 5'-terminal nucleotides of STMV RNA had greater than 65% sequence similarity with the 16 5'-terminal nucleotides of brome mosaic virus (RNA 3 and 50% sequence similarity with the 12 5'-terminal nucleotides of the Q strain of cucumber mosaic virus RNA 3. The first open reading frame (ORF) beginning at base 53 encoded a 6800 Mr protein that corresponded in size to a major in vitro translation product directed by STMV RNA. A second ORF, beginning at nucleotide 163, had the capacity to code for a protein that corresponded in size (17,500 Mr) to the other major in vitro translation product. The first 12 codons of this ORF corresponded to the sequence of the N-terminal amino acids of the capsid protein. Western-blot analysis of the in vitro translation products revealed that the 17,500 Mr protein had the same electrophoretic mobility as the authentic capsid protein; it was also antigenically related to the capsid protein, but the 6800 Mr protein was not. Time course analysis of in vitro translation demonstrated that the 6800 Mr protein was synthesized at the same time as the capsid protein and did not arise by the proteolytic cleavage of a larger precursor polypeptide. These results suggest that the genome of STMV functioned as a polycistronic messenger RNA. It has not been determined if the 6800 Mr protein is synthesized in vivo. STMV RNA had untranslated regions of 52 and 418 nucleotides at its 5' and 3' termini, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)


Preconditioning by light-load eccentric exercise is equally effective as low-level laser therapy in attenuating exercise-induced muscle damage in collegiate men.
Previous studies have already reported an independent effect of light-load eccentric exercise (10% eccentric exercise contraction [EEC]) and low-level laser therapy (LLLT) as a protective measure against more strenuous eccentric exercise. However, the difference between these two interventions is largely unknown. Therefore, the present study aimed to compare the preconditioning effect of 10% EEC vs. LLLT on subjective, physiological, and biochemical markers of muscle damage in elbow flexors in collegiate men. All 36 enrolled subjects were randomly assigned to either 10% EEC or LLLT group. Subjects in 10% EEC group performed 30 repetitions of an eccentric exercise with 10% maximal voluntary contraction strength 2 days prior to maximal eccentric exercise bout, whereas subjects in LLLT group were given LLLT. All the indirect markers of muscle damage were measured pre-exercise and at 24, 48, and 72 hours after the exercise-induced muscle damage protocol. The muscle soreness was reduced in both groups (p = 0.024); however, soreness was attenuated more in LLLT group at 48 hours (33.5 vs. 42.7, p = 0.004). There was no significant difference between the effect of 10% EEC and LLLT groups on other markers of muscle damage like a maximum voluntary isometric contraction (p = 0.47), range of motion (p = 0.16), upper arm circumference (p = 0.70), creatine kinase (p = 0.42), and lactate dehydrogenase (p = 0.08). Within-group analysis showed both interventions provided similar protection over time. This study indicated that light-load eccentric exercise confers similar protective effect against subsequent maximal eccentric exercise as LLLT. Both the treatments could be used reciprocally based on the patient preference, costs, and feasibility of the equipment.


Trophic structure of fish communities in mangrove systems subject to different levels of anthropogenic intervention, Tropical Eastern Pacific, Colombia.
Mangroves are important ecosystems of tropical and subtropical shorelines. Anthropogenic activity decreases their habitat quality, affecting structural and functional trophic features. We hypothesized that higher levels of anthropogenic intervention generate diversity loss and modify the trophic structure of tropical mangrove fish communities. We compared the taxonomic and isotopic (δ13C/δ15N) composition, abundance, trophic position, and isotopic niche of fish communities from three mangrove systems with different anthropogenic intervention levels in the Colombian Pacific. Non-parametric statistical tests and a Bayesian approach were used to analyze data. A total of 1254 specimens belonging to 23 families, 25 genera, and 30 species were identified, presenting higher abundance (821) in moderate anthropogenic intervention level mangrove (Moderate-AIL), with high dominance of one species (Lile stolifera). The low anthropogenic intervention level mangrove (Low-AIL) was the second in abundance (291) but exhibited a greater number of species (23), while the high anthropogenic intervention level mangrove (High-AIL) presented the least abundance (142) and species number (17). The isotopic composition reveals that Moderate and High-AIL mangroves presented enriched 13C and 15 N compared to Low-AIL (~ 2 to 4 ‰). Mean trophic position (TP) of communities was slightly higher in the more intervened systems (~ 1 to 2 orders of magnitude), as well as in specific species (Centropomus spp.). Isotopic niche width (TA and SEAc) was greater in High-AIL (41.1 and 9.2), more than doubled compared to Moderate-AIL (33.0 and 4.1). In High-AIL isotopic niche width increased, indicating lesser availability of prey and basal resources. The results obtained in this study support the proposed hypothesis and, suggest that anthropogenic intervention modifies diversity and food webs dynamics, affecting the transfer of matter and energy from macrotidal tropical mangroves to coastal ecosystems. However, it is recommended to be careful concluding differences based exclusively on the anthropogenic intervention level, since it is widely documented that mangrove settings also influence the analyzed trophic parameters.


Hospital care in severe trauma: Initial strategies and life-saving surgical procedures.
Severe trauma patients should be received at the hospital by a multidisciplinary team directed by a "trauma leader" and all institutions capable of receiving such patients should be well organized. As soon as the patient is accepted for care, the entire team should be prepared so that there is no interruption in the pre-hospital chain of care. All caregivers should thoroughly understand the pre-established protocols of diagnostic and therapeutic strategies to allow optimal management of unstable trauma victims in whom hemostasis must be obtained as soon as possible to decrease the morbid consequences of post-hemorrhagic shock. In patients with acute respiratory, circulatory or neurologic distress, several surgical procedures must be performed without delay by whichever surgeon is on call. Our goal is to describe these salvage procedures including invasive approaches to the upper respiratory tract, decompressive thoracostomy, hemostatic or resuscitative thoracotomy, hemostatic laparotomy, preperitoneal pelvic packing, external pelvic fixation by a pelvi-clamp, decompressive craniotomy. All of these procedures can be performed by all practitioners but they require polyvalent skills and training beforehand.


Relationship between motor learning and gambling propensity in Parkinson's disease.
The basal ganglia and related dopaminergic cortical areas are important neural systems underlying motor learning and are also implicated in impulse control disorders (ICDs). Motor learning impairments and ICDs are frequently observed in Parkinson's disease (PD). Nevertheless, the relationship between motor learning ability and ICDs has not been elucidated. We examined the relationship between motor learning ability and gambling propensity, a possible symptom for prodromal ICDs, in PD patients. Fifty-nine PD patients without clinical ICDs and 43 normal controls (NC) were administered a visuomotor rotation perturbation task and the Iowa Gambling Task (IGT) to evaluate motor learning ability and gambling propensity, respectively. Participants also performed additional cognitive assessments and underwent brain perfusion SPECT imaging. Better motor learning ability was significantly correlated with lower IGT scores, i.e., higher gambling propensity, in PD patients but not in NC. The higher scores on assessments reflecting prefrontal lobe function and well-preserved blood perfusion in prefrontal areas were correlated with lower IGT scores along with better motor learning ability. Our findings suggest that better motor learning ability and higher gambling propensity are based on better prefrontal functions, which are in accordance with the theory that the prefrontal cortex is one of the common essential regions for both motor learning and ICDs.


Dementia of the Alzheimer type: effects on the spontaneous EEG described by dipole sources.
The present study on brain electrical activity in healthy subjects (n = 35) and patients suffering from dementia of the Alzheimer type (DAT; n = 35) used Fast Fourier Transformation-dipole approximation to quantify differences between the two groups. DAT patients showed a shift of alpha and beta activity toward frontal brain regions. The amount of this shift correlated with the degree of dementia. The relative distribution of magnitude of activity between the frequency bands differed between DAT patients and control subjects. DAT patients had higher magnitudes in the slow frequency range, correlating with the severity of dementia, and lower ones in the alpha and beta range compared with findings in age-matched control subjects.


Rb1 negatively regulates bone formation and remodeling through inhibiting transcriptional regulation of YAP in Glut1 and OPG expression and glucose metabolism in male mice.
Bone is a highly dynamic organ that undergoes constant bone formation and remodeling, and glucose as a major nutrient is necessary for bone formation and remodeling. Retinoblastoma (Rb1) is a critical regulator of mesenchymal stem cells (MSCs) fate, but how Rb1 regulates bone formation and remodeling is poorly understood. We generated MSCs- and osteoprogenitors-specific Rb1 knockout mouse models and utilized these models to explore the function and mechanism of Rb1 in regulating bone formation and remodeling in vivo and in vitro primary cell culture. Rb1 deficiency in MSCs significantly increased bone mass and impaired osteoclastogenesis. Consistently, depletion of Rb1 in osteoprogenitors significantly promoted bone formation. Mechanistically, loss of Rb1 in MSCs elevated YAP nuclear translocation and transcriptional activity of YAP/TEAD1 complex, thereby increasing the transcriptional expression of Glut1 and OPG. Moreover Prx1-Cre; Rb1f/f mice displayed hypoglycemia with increased systemic glucose tolerance instead of increased insulin level. In vitro data revealed that Rb1-mutant MSCs enhanced glucose uptake and lactate and ATP production. Increased osteogenesis caused by increased glucose metabolism and decreased osteoclastogenesis caused by increased expression of OPG eventually resulted in increased bone formation and remodeling. Collectively, these findings demonstrated that Rb1 in MSCs inhibits YAP-medicated Glut1 and OPG expression to control glucose metabolism, osteogenesis and osteoclastogenesis during bone formation and remodeling, which provide new insights that controlling Rb1 signaling may be a potential strategy for osteopetrosis.


Schmallenberg virus infection in South American camelids: Field and experimental investigations.
During the first epizootic wave of the novel, teratogenic Schmallenberg virus (SBV, Orthobunyavirus) in ruminants in Northern Europe, serological evidence of a previous SBV-infection demonstrated that South American camelids (SAC) are also susceptible to SBV. However, their potential role in SBV spread remains unknown. To investigate the prevalence and course of SBV-infection in SAC, a German field study and an animal trial with three llamas and three alpacas were conducted. From September 2012 to December 2013, 313 of 502 SAC (62.35%) were found SBV seropositive, but negative for SBV-RNA. The estimated between-district (94.23% of 52) and median within-district (71.43%) and herd (73.13%) SBV seroprevalence in German SAC was similar to the seroprevalence reported in cattle herds and sheep flocks at the time. An age of >1 year was found a statistically significant risk factor for SBV-infection, which could be explained by the spatio-temporal spread of SBV in Germany during the study period. No clinical signs or an increase of abortion and congenital malformation associated with SBV-infection in SAC were reported by the study participants. Similar to SBV-infected ruminants, SBV-RNAemia in experimentally SBV-infected SAC was detected for a short time between days 3 and 7 after infection (dpi), and seroconversion occurred between 9 and 21 dpi. Despite the similar virological and serological results, the lack of clinical signs and congenital malformation associated with SBV-infection suggests that SBV causes subclinical infection in SAC. However, their role as reservoirs in the spread of SBV has to be further investigated.


Long-term impact of arsenic in drinking water on bladder cancer health care and mortality rates 20 years after end of exposure.
In this study we assessed bladder cancer health care and mortality trends in recent decades in a well studied arsenic exposed area in Northern Chile. Arsenic levels in the affected region were obtained for the last 60 years, and correlated with bladder cancer hospital discharge and mortality rates in recent decades. Bladder cancer hospital discharge rates were significantly higher in the affected region (peak RR 3.6, 95% CI 3.0-4.7). Mortality rates for bladder cancer showed a trend of increase during the period analyzed, reaching peak mortality rates of 28.4 per 100,000 for men and 18.7 per 100,000 for women in the last 10 years. Poisson regression models showed an increased mortality risk in the studied region compared to the rest of the country until the present for men (IRR 5.3, 95% CI 4.8-5.8) and women (IRR 7.8, 95% CI 7.0-8.7). Mean age at cancer specific death was significantly lower in the exposed region (69.6 years, 95% CI 68.4-70.7 vs 73.7 years, 95% CI 73.3-74.2, p <0.01). Exposure to arsenic is related to a significant need for bladder cancer health care and to high mortality rates even 20 years after having controlled arsenic levels in drinking water. Affected individuals should be aware of the significant impact of this ecological factor. Further research is required to identify strategies for the management of bladder cancer in arsenic exposed populations.


[Importance of the friction between plate and bone in the anchoring of plates for osteosynthesis. Determination of the coefficient of metal-bone friction in animal in vivo].
In mechanical devices in general the friction plays an important role in force transmission. To find out about the magnitude of frictional forces between internal fixation plate and bone in vivo, simultaneous strain recording in plated bone and instrumented plate were performed. After 3 weeks of implantation the torque applied to a single plate-screw was set to definite levels. Thus it could be determined at what level of screw compression and at what tangential load of the related surface bone-plate, gliding occurred. The coefficient of friction was found to be 0.2 +/- 0.09 (x +/- Sx). It can be concluded that in vivo friction produced at the plate-bone contact surface is able to guarantee stability of fixation.


Identification of a potent DNase activity associated with RNase T of Escherichia coli.
RNase T was first identified as an enzyme responsible for end turnover of tRNA in Escherichia coli. Its activity, specific for tRNA-C-C-A, catalyzes the release of tRNA-C-C and AMP. RNase T, along with several other RNases, plays a role in maturation of several other RNA species by a similar limited nuclease activity. In previous work, we identified the gene for RNase T, rnt, as a high copy suppressor of the UV sensitivity conferred by deficiency in three single-strand DNA-specific exonucleases, RecJ, exonuclease I, and exonuclease VII. This suggested that RNase T may process DNA substrates as well. In this work, we show that purified RNase T possesses a potent 3' to 5' single-strand DNA-specific exonucleolytic activity. Its Km for single-strand DNA substrates is many orders of magnitude lower than that for tRNA, suggesting that single-strand DNA may be a natural biological substrate for RNase T. We suggest that the DNase activity of RNase T may play a role in end trimming reactions during DNA recombination and/or DNA repair.


Fructose-1,6-bisphosphate reduces the mortality in Candida albicans bloodstream infection and prevents the septic-induced platelet decrease.
Due to the fact that an increased number of patients have experienced bloodstream infections caused by Candida species and the high mortality of this infection, there is a need for a strategy to reduce this scenery. One possible strategy is the use of new drugs, such as fructose-1,6-bisphosphate (FBP), which is a high-energy glycolytic metabolite and has shown to have therapeutic effects in several pathological conditions such as ischemia, shock, toxic injuries, and bacterial sepsis. The aim of this manuscript was to determine the role of FBP in experimental Candida albicans bloodstream infection. We used mice that were divided into three experimental groups: sham (not induced), bloodstream infection (induced with intratracheal instillation of C. albicans) and FBP (bloodstream infection plus FBP 500 mg/kg i.p.). Blood was taken for assessment of complete hematological profile and cytokine assay (IL-6 and MCP-1). Results of the study demonstrated that mortality decreased significantly in groups that received FBP. All cytokine and hematological indexes of FBP group were similar to bloodstream infection group with exception of platelets count. FBP significantly prevented the decrease in platelets. Taken together, our results demonstrate that FBP prevented the mortality in C. albicans bloodstream infection.


Behavioral and pharmacological assessment of butyrylcholinesterase in rats.
Advances in the treatment of organophosphorus (OP) toxicity have focussed on the use of exogenous cholinesterases to act as scavengers for the OP agent. To further investigate the feasibility of the scavenger approach, we evaluated the effects of highly purified horse serum butyrylcholinesterase (HS-BChE) on performance in rats. HS-BChE (5000 U, IP) produced substantial increases in blood enzyme activity for up to 72 h after injection. HS-BChE (5000 U, IP) had no effect on acquisition or retention of a passive avoidance task. In contrast, atropine sulfate (10 mg/kg) impaired retention when tested 168 h after administration. When examined for 10 days following administration, HS-BChE (7500 U, IP) had no effect on either total daily motor activity or circadian pattern of activity. HS-BChE (5000 U, IM) also had no acute or prolonged effects on the rate of lever pressing maintained by a VI56 s schedule of food reinforcement. HS-BChE (7500 U, IM) was observed to confer significant, but partial, protection against response rate decreases produced by the OP, MEPQ, under the VI56 s schedule of reinforcement. These results suggest that, in rats, HS-BChE, at doses that attenuate OP toxicity, may be devoid of cognitive or motor effects.


The six Tomato yellow leaf curl virus genes expressed individually in tomato induce different levels of plant stress response attenuation.
Tomato yellow leaf curl virus (TYLCV) is a begomovirus infecting tomato plants worldwide. TYLCV needs a healthy host environment to ensure a successful infection cycle for long periods. Hence, TYLCV restrains its destructive effect and induces neither a hypersensitive response nor cell death in infected tomatoes. On the contrary, TYLCV counteracts cell death induced by other factors, such as inactivation of HSP90 functionality. Suppression of plant death is associated with the inhibition of the ubiquitin 26S proteasome degradation and with a deactivation of the heat shock transcription factor HSFA2 pathways (including decreased HSP17 levels). The goal of the current study was to find if the individual TYLCV genes were capable of suppressing HSP90-dependent death and HSFA2 deactivation. The expression of C2 (C3 and CP to a lesser extent) caused a decrease in the severity of death phenotypes, while the expression of V2 (C1 and C4 to a lesser extent) strengthened cell death. However, C2 or V2 markedly affected stress response under conditions of viral infection. The downregulation of HSFA2 signaling, initiated by the expression of C1 and V2, was detected in the absence of virus infection, but was enhanced in infected plants, while CP and C4 mitigated HSFA2 levels only in the infected tomatoes. The dependence of analyzed plant stress response suppression on the interaction of the expressed genes with the environment created by the whole virus infection was more pronounced than on the expression of individual TYLCV genes.


Protective action of 17beta-estradiol in cardiac cells: implications for hyperkalemic cardioplegia.
Hyperkalemic cardioplegic solutions effectively arrest the heart, but may also induce intracellular Ca2+ loading and cellular hypercontracture, which could contribute to ventricular dysfunction associated with global surgical ischemia. Recently, it has been proposed that 17beta-estradiol may possess protective properties in the ischemic myocardium. The purpose of the present study was to examine the action of 17beta-estradiol on cardiac cells exposed to hyperkalemic stress. Single ventricular cardiomyocytes, a preparation devoid of vascular and neuronal elements, were isolated from guinea pig hearts, loaded with a Ca2+-sensitive fluorescent probe, and imaged by digital epifluorescent microscopy. The emitted fluorescence of the probe, a measure of intracellular Ca2+ concentration, and cell length were simultaneously recorded during hyperkalemic challenge, in the absence or presence of 17beta-estradiol. In control cardiomyocytes, the cytosolic concentration of Ca2+ was 138+/-11 nmol/L and cell length 93+/-11 microm. Exposure to high K+ (+16 mmol/L KCl) significantly increased cytosolic Ca2+ to 2,191+/-87 nmol/L (p < 0.001), and produced cell shortening (length at 39+/-5 microm; p < 0.001). 17beta-Estradiol (10 micromol/L) acutely prevented high K+ to induce either intracellular Ca2+ loading (144+/-13 nmol/L, p < 0.001) or hypercontracture (91+/-10 microm, p < 0.001). Tamoxifen (10 micromol/L), an antiestrogen, abolished the protective effect of 17beta-estradiol. We conclude that 17beta-estradiol prevents hyperkalemia-induced Ca2+ loading and hypercontracture through a direct and tamoxifen-sensitive action in cardiomyocytes. This study raises the possibility that 17beta-estradiol should be considered as a cardioprotective adjunct toward a safer hyperkalemic cardioplegia.


Transcriptomic complexity of the human malaria parasite Plasmodium falciparum revealed by long-read sequencing.
The Plasmodium falciparum human malaria parasite genome is incompletely annotated and does not accurately represent the transcriptomic diversity of this species. To address this need, we performed long-read transcriptomic sequencing. 5' capped mRNA was enriched from samples of total and nuclear-fractionated RNA from intra-erythrocytic stages and converted to cDNA library. The cDNA libraries were sequenced on PacBio and Nanopore long-read platforms. 12,495 novel isoforms were annotated from the data. Alternative 5' and 3' ends represent the majority of isoform events among the novel isoforms, with retained introns being the next most common event. The majority of alternative 5' ends correspond to genomic regions with features similar to those of the reference transcript 5' ends. However, a minority of alternative 5' ends showed markedly different features, including locations within protein-coding regions. Alternative 3' ends showed similar features to the reference transcript 3' ends, notably adenine-rich termination signals. Distinguishing features of retained introns could not be observed, except for a tendency towards shorter length and greater GC content compared with spliced introns. Expression of antisense and retained intron isoforms was detected at different intra-erythrocytic stages, suggesting developmental regulation of these isoform events. To gain insights into the possible functions of the novel isoforms, their protein-coding potential was assessed. Variants of P. falciparum proteins and novel proteins encoded by alternative open reading frames suggest that P. falciparum has a greater proteomic repertoire than the current annotation. We provide a catalog of annotated transcripts and encoded alternative proteins to support further studies on gene and protein regulation of this pathogen.


Pituitary metastasis from renal cell carcinoma presenting with significant hyperprolactinemia, case report.
Kidney metastasis to the pituitary gland can cause hyperprolactinemia even above 250 ng/mL. Although the treatment of metastasis is palliative, surgical decompression could play a major role in the recovery of symptoms and improve quality of life. Pituitary metastasis should be considered in the evaluation of an unusual pituitary mass. Pituitary tumors are frequently encountered in the neurosurgical setting. Although the majority of them are pituitary adenomas, rare entities encompass pituitary metastasis. They should be differentiated from pituitary adenomas because their management and prognosis are different. We report a 53-year-old female who complained of headache and had remarkable hyperprolactinemia (271.1 ng/mL). Having considered macroprolactinoma as the initial diagnosis, medical treatment was initiated with Cabergoline. Subsequently, the patient's vision deteriorated which prompted us to perform endoscopic endonasal transsphenoidal surgery. Histologic examination of the resected tumor revealed metastatic renal cell carcinoma. Main treatment for these subjects is palliative; and unlike the pituitary adenoma, the prognosis is unfortunately poor. Pituitary metastasis should be considered in the evaluation of an unusual pituitary mass associated with hyperprolactinemia.


A New Hydrogen-Producing Strain and Its Characterization of Hydrogen Production.
A newly isolated photo non-sulfur (PNS) bacterium was identified as Rhodopseudomonas palustris PB-Z by sequencing of 16S ribosomal DNA (rDNA) genes and phylogenetic analysis. Under vigorous stirring (240 rpm), the hydrogen production performances were greatly improved: The maximum hydrogen production rate and cumulative hydrogen production increased by 188.9 ± 0.07 % and 83.0 ± 0.06 %, respectively, due to the hydrogen bubbles were immediately removed from the culture medium. The effects of different wavelength of light on hydrogen production with stirring were much different from that without stirring. The ranking on the photo-hydrogen production performance was white > yellow > green > blue > red without stirring and white > yellow > blue > red > green under stirring. The best light source for hydrogen production was tungsten filament lamp. The optimum temperature was 35 °C. The maximal hydrogen production rate and cumulative hydrogen production reached 78.7 ± 2.3 ml/l/h and 1728.1 ± 92.7 mol H2/l culture, respectively, under 35 °C, 240 rpm, and illumination of 4000 lux. Pyruvate was one of the main sources of CO2 and has a great impact on the gas composition.


Order statistics theory of unfolding of multimeric proteins.
Dynamic force spectroscopy has become indispensable for the exploration of the mechanical properties of proteins. In force-ramp experiments, performed by utilizing a time-dependent pulling force, the peak forces for unfolding transitions in a multimeric protein (D)(N) are used to map the free energy landscape for unfolding for a protein domain D. We show that theoretical modeling of unfolding transitions based on combining the observed first (f(1)), second (f(2)), …, N(th) (f(N)) unfolding forces for a protein tandem of fixed length N, and pooling the force data for tandems of different length, n(1) <n(2) < … <N, leads to an inaccurate estimation of the distribution of unfolding forces for the protein D, ψ(D)(f). This problem can be overcome by using Order statistics theory, which, in conjunction with analytically tractable models, can be used to resolve the molecular characteristics that determine the unfolding micromechanics. We present a simple method of estimation of the parent distribution, ψ(D)(f), based on analyzing the force data for a tandem (D)(n) of arbitrary length n. Order statistics theory is exemplified through a detailed analysis and modeling of the unfolding forces obtained from pulling simulations of the monomer and oligomers of the all-β-sheet WW domain.


Participant preferences for the provision of registration trials results.
Clinical trials leading to drug approval (registration trials) play a central role in the drug development process, and attention has recently been paid to providing trial results to participants. In the present study, we examined the preferences of participants of registration trials for the provision of trial-related information. We used questionnaires to survey the preferences of registration trial participants at Tokushima University Hospital and Tokushima National Hospital. Of the 15 questions, 6 related to participant characteristics and the trials in which they participated, while 9 questions were concerned with preferences for the provision of information. A five-point scale (strongly agree, agree, neutral, disagree, and strongly disagree) was used, and positive answers (strongly agree and agree) were considered to indicate a positive preference. Of the 58 subjects, 1 declined, giving a response rate of 98%. More than 70% of participants preferred to obtain information, even if they had served as controls. More than 80% of participants agreed to obtain information relating to trial results, even if the results were negative, and more than 80% of participants agreed to obtain information on the labeling state of the agent, even if development had ceased. Although more than 60% of participants agreed for the provision of information on their allocation and around more than 70% agreed to the provision of information on registration trials status, significantly fewer participants with difficult-to-treat diseases (for example, neurological and malignant diseases) agreed to obtain information compared with participants with other types of diseases (for example, acute, chronic, and psychological diseases). More than 50% of participants desired information to be provided directly by the physician, while a considerable number of participants desired information by means of clinical research coordinators (CRCs) (24.4%) or by posted letter (33.3%). The present results suggest the preferences for the provision of individual and overall information concerning research results. However, further study is warranted to determine participant preferences more precisely and the effect of the CRC-initiated infrastructure for providing information on patient satisfaction and for promoting registration trials.


Investigation of anal sphincter function following Mainz pouch type II urinary diversion after radical cystectomy.
Our aim was to evaluate the anal sphincter function following cystectomy with urinary diversion of Mainz pouch II. Seventy-six patients were involved in our survey, and the cohort was for two groups divided. The first group was a retrospective review of 40 patients with examination of the state of continence. Comparative examinations on anal sphincter function and the quality of life survey were carried out. The second group consisting of 15 patients underwent a prospective investigation including rectal manometry in both the pre- and postoperative periods. Measurements of resting anal sphincter pressure (RASP), maximal anal closing pressure (MACP) and the function of the recto anal inhibitions reflex were taken. In the first part of our investigation, 80% of the patients were considered as continent. There were no significant differences observed between RASP values in the cases of continent as well as of incontinent patients (79.2 ± 2 vs. 73.6 ± 68.4 mmHg, p = 0-53); however, the MACP values of the continent patients were significantly higher (204.3 ± 22.8 vs. 117.3 ± 14 mmHg, p = 0.001). In the course of the second experiment, both the RASP (86.3 ± 18.7 vs. 76.1 ± 13.9 mmHg p = 0.0049) and the MACP (232.2 ± 53.8 vs. 194.1 ± 74.5 mmHg, p = 0.0054) were detected as decreasing in the case of the incontinent group. A decrease in rectal sphincter function is responsible for incontinence following Mainz pouch type II diversion, and this dysfunction can be correlated with the surgery. Ureterosigmoideostomy is therefore considered as a useful method of urinary diversion only in selected cases with proven good sphincter function.


Uniportal Video-Assisted Thoracic Surgery Esophagectomy.
Several minimally invasive approaches for esophagectomy have been described, including robot-assisted esophagectomy and hybrid techniques, total transhiatal laparoscopic approach, esophagectomy using right thoracoscopy, combined laparoscopic and right thoracoscopic esophagectomy, and esophageal resection through mediastinoscopy. However, very few publications have focused on the uniportal video-assisted thoracic surgery (VATS) approach. The authors describe their technique of the minimally invasive esophagectomy using uniportal VATS as the thoracic step.


An ancient defense mechanism: Conservation of gasdermin-mediated pyroptosis.
The gasdermins are a family of pore-forming proteins involved in various cellular processes such as cell death and inflammation. A new study in PLOS Biology explores the evolutionary history of gasdermins across metazoans, highlighting the conservation and divergence of gasdermin E.