Tyrosines in the carboxyl terminus regulate Syk kinase activity and function.
The Syk tyrosine kinase family plays an essential role in immunoreceptor tyrosine-based activation motif (ITAM) signaling. The binding of Syk to tyrosine-phosphorylated ITAM subunits of immunoreceptors, such as FcepsilonRI on mast cells, results in a conformational change, with an increase of enzymatic activity of Syk. This conformational change exposes the COOH-terminal tail of Syk, which has three conserved Tyr residues (Tyr-623, Tyr-624, and Tyr-625 of rat Syk). To understand the role of these residues in signaling, wild-type and mutant Syk with these three Tyr mutated to Phe was expressed in Syk-deficient mast cells. There was decreased FcepsilonRI-induced degranulation, nuclear factor for T cell activation and NFkappaB activation with the mutated Syk together with reduced phosphorylation of MAP kinases p38 and p42/44 ERK. In non-stimulated cells, the mutated Syk was more tyrosine phosphorylated predominantly as a result of autophosphorylation. In vitro, there was reduced binding of mutated Syk to phosphorylated ITAM due to this increased phosphorylation. This mutated Syk from non-stimulated cells had significantly reduced kinase activity toward an exogenous substrate, whereas its autophosphorylation capacity was not affected. However, the kinase activity and the autophosphorylation capacity of this mutated Syk were dramatically decreased when the protein was dephosphorylated before the in vitro kinase reaction. Furthermore, mutation of these tyrosines in the COOH-terminal region of Syk transforms it to an enzyme, similar to its homolog ZAP-70, which depends on other tyrosine kinases for optimal activation. In testing Syk mutated singly at each one of the tyrosines, Tyr-624 but especially Tyr-625 had the major role in these reactions. Therefore, these results indicate that these tyrosines in the tail region play a critical role in regulating the kinase activity and function of Syk.


Minimal Heating at the Skin Surface During Transcranial Direct Current Stimulation.
To assess if transcranial direct current stimulation (tDCS) produces a temperature change at the skin surface, if any change is stimulation polarity (anode or cathode) specific, and the contribution of passive heating (joule heat) or blood flow on such change. Temperature differences (ΔTs) in an agar phantom study and an in vivo study (forearm stimulation) including 20 volunteers with both experimental measures and finite element method (FEM) multiphysics prediction (current flow and bioheat) models of skin comprising three tissue layers (epidermis, dermis, and subcutaneous layer with blood perfusion) or of the phantom for active stimulation and control cases were compared. Temperature was measured during pre, post, and stimulation phases for both phantom and subject's forearms using thermocouples. In the phantom, ΔT under both anode and cathode, compared to control, was not significantly different and less than 0.1°C. Stimulation of subjects resulted in a gradual increase in temperature under both anode and cathode electrodes, compared to control (at t = 20 min: ΔTanode = 0.9°C, ΔTcathode = 1.1°C, ΔTcontrol = 0.05°C). The FEM phantom model predicted comparable maximum ΔT of 0.27°C and 0.28°C (at t = 20 min) for the control and anode/cathode cases, respectively. The FEM skin model predicted a maximum ΔT at t = 20 min of 0.98°C for control and 1.36°C under anode/cathode electrodes. Taken together, our results indicate a moderate and nonhazardous increase in temperature at the skin surface during 2 mA tDCS that is independent of polarity, and results from stimulation induced blood flow rather than joule heat.


[Zygomycosis following liver transplantation in adults: report of three cases].
We report three cases of zygomycosis following liver transplant in a series of 300 patients. Diagnosis was determined via anatomicopathological examination (on necropsy in two cases and during surgery in one case). The disease had different manifestations: rhinomaxillary, gastrointestinal and, in one case, it compromised the liver artery anastomosis. In this case, surgical removal of the affected region and use of amphotericin B achieved resolution.


Infectivity of Lactobacillus rhamnosus and Lactobacillus paracasei isolates in a rat model of experimental endocarditis.
The potential pathogenicity of selected (potentially) probiotic and clinical isolates of Lactobacillus rhamnosus and Lactobacillus paracasei was investigated in a rat model of experimental endocarditis. In addition, adhesion properties of the lactobacilli for fibrinogen, fibronectin, collagen and laminin, as well as the killing activity of the platelet-microbicidal proteins fibrinopeptide A (FP-A) and connective tissue activating peptide 3 (CTAP-3), were assessed. The 90 % infective dose (ID(90)) of the L. rhamnosus endocarditis isolates varied between 10(6) and 10(7) c.f.u., whereas four of the six (potentially) probiotic L. rhamnosus isolates showed an ID(90) that was at least 10-fold higher (10(8) c.f.u.) (P<0.001). In contrast, the two other probiotic L. rhamnosus isolates exhibited an ID(90) (10(6) and 10(7) c.f.u.) comparable to the ID(90) of the clinical isolates of this species investigated (P>0.05). Importantly, these two probiotic isolates shared the same fluorescent amplified fragment length polymorphism cluster type as the clinical isolate showing the lowest ID(90) (10(6) c.f.u.). L. paracasei tended to have a lower infectivity than L. rhamnosus (ID(90) of 10(7) to > or =10(8) c.f.u.). All isolates had comparable bacterial counts in cardiac vegetations (P>0.05). Except for one L. paracasei strain adhering to all substrates, all tested lactobacilli adhered only weakly or not at all. The platelet peptide FP-A did not show any microbicidal activity against the tested lactobacilli, whereas CTAP-3 killed the majority of the isolates. In general, these results indicate that probiotic lactobacilli display a lower infectivity in experimental endocarditis compared with true endocarditis pathogens. However, the difference in infectivity between L. rhamnosus endocarditis and (potentially) probiotic isolates could not be explained by differences in adherence or platelet microbicidal protein susceptibility. Other disease-promoting factors may exist in these organisms and warrant further investigation.


Nominal group technique: an effective method for obtaining group consensus.
This paper aims to demonstrate the versatility and application of nominal group technique as a method for generating priority information. Nominal group technique was used in the context of four focus groups involving clinical experts from the emergency department (ED) and obstetric and midwifery areas of a busy regional hospital to assess the triage and management of pregnant women in the ED. The data generated were used to create a priority list of discussion triggers for the subsequent Participatory Action Research Group. This technique proved to be a productive and efficient data collection method which produced information in a hierarchy of perceived importance and identified real world problems. This information was vital in initiating the participatory action research project and is recommended as an effective and reliable data collection method, especially when undertaking research with clinical experts.


The coactivator TIF2 contains three nuclear receptor-binding motifs and mediates transactivation through CBP binding-dependent and -independent pathways.
The nuclear receptor (NR) coactivator TIF2 possesses a single NR interaction domain (NID) and two autonomous activation domains, AD1 and AD2. The TIF2 NID is composed of three NR-interacting modules each containing the NR box motif LxxLL. Mutation of boxes I, II and III abrogates TIF2-NR interaction and stimulation, in transfected cells, of the ligand-induced activation function-2 (AF-2) present in the ligand-binding domains (LBDs) of several NRs. The presence of an intact NR interaction module II in the NID is sufficient for both efficient interaction with NR holo-LBDs and stimulation of AF-2 activity. Modules I and III are poorly efficient on their own, but synergistically can promote interaction with NR holo-LBDs and AF-2 stimulation. TIF2 AD1 activity appears to be mediated through CBP, as AD1 could not be separated mutationally from the CBP interaction domain. In contrast, TIF2 AD2 activity apparently does not involve interaction with CBP. TIF2 exhibited the characteristics expected for a bona fide NR coactivator, in both mammalian and yeast cells. Moreover, in mammalian cells, a peptide encompassing the TIF2 NID inhibited the ligand-induced AF-2 activity of several NRs, indicating that NR AF-2 activity is either mediated by endogenous TIF2 or by coactivators recognizing a similar surface on NR holo-LBDs.


Wavelength-dependent model of Kr flash lamp emission and absorption.
We present a simple model of the angular intensity distribution of linear flash lamps for laser pumping application, which also takes into account the plasma column absorption during flash pulse. The model parameters are derived by single-shot measurements of plasma emission and transmission, performed with an imaging spectrometer and a CCD detector. The model consists in a linear superposition, wavelength by wavelength, of a surface (opaque) and a volume (transparent) emitter and represents a good approximation of the time-averaged lamp emission and absorption for pulse durations from 50 mus to 2 ms. This model is a suitable tool for reflector optimization of flash-lamp-pumped solid-state lasers, allowing complete wavelength-dependent ray tracing with currently available computing power. Some sample applications are also shown and discussed, with results of ray tracing for a slab laser and evaluation of the effects of lamp blackbody emission for several laser active media and flash lamp excitation pulses.


CD4 segregates into specific detergent-resistant T-cell membrane microdomains.
In T cells, glycolipids, glycoproteins attached to the membrane via a glycosylphosphatidylinositol (GPI) anchor, and Src-like tyrosine kinases are highly enriched in a membrane fraction resistant to solubilization by nonionic detergents. We have investigated the distribution of CD4 in T-cell membranes and found that approximately 10% of the CD4 co-receptor is associated with detergent-insoluble membrane microdomains, whilst the remaining 90% is in soluble membranes. Moreover, approximately 60% of the "insoluble CD4" is present in membrane microdomains containing GPI-anchored proteins and high glycolipid-dependent kinase activity, whereas the remaining 40% displays no association with GPI-anchored proteins and lacks glycolipid-associated kinase activity These results indicate that CD4 segregates at least into three different membrane microenvironments: 1) soluble membranes; 2) insoluble membrane microdomains containing GPI-anchored proteins; and 3) insoluble membrane microdomains devoid of GPI-anchored proteins. The level of CD4 in insoluble membranes was not modified upon triggering activation by T-cell receptor-crosslinking but detectable amounts of CD3 subunits were recruited into these specialized membranes under those conditions. The physical separation of CD4 into different membrane microenvironments raises the possibility of that some of the multiple functions of CD4 might segregate into distinct types of lipid microenvironment. The fact that components of T-cell receptor/CD3 complex were recruited into insoluble membranes upon stimulation is consistent with the CD4 present in this membrane fraction might participate in T-cell receptor-triggered activation events.


Activation of the glutamatergic cingulate cortical-cortical connection facilitates pain in adult mice.
The brain consists of the left and right cerebral hemispheres and both are connected by callosal projections. Less is known about the basic mechanism of this cortical-cortical connection and its functional importance. Here we investigate the cortical-cortical connection between the bilateral anterior cingulate cortex (ACC) by using the classic electrophysiological and optogenetic approach. We find that there is a direct synaptic projection from one side ACC to the contralateral ACC. Glutamate is the major excitatory transmitter for bilateral ACC connection, including projections to pyramidal cells in superficial (II/III) and deep (V/VI) layers of the ACC. Both AMPA and kainate receptors contribute to synaptic transmission. Repetitive stimulation of the projection also evoked postsynaptic Ca2+ influx in contralateral ACC pyramidal neurons. Behaviorally, light activation of the ACC-ACC connection facilitated behavioral withdrawal responses to mechanical stimuli and noxious heat. In an animal model of neuropathic pain, light inhibitory of ACC-ACC connection reduces both primary and secondary hyperalgesia. Our findings provide strong direct evidence for the excitatory or facilitatory contribution of ACC-ACC connection to pain perception, and this mechanism may provide therapeutic targets for future treatment of chronic pain and related emotional disorders.


Progress in preservation of intestinal grafts by oxygenated hypothermic machine perfusion.
Intestine transplantation (IT) is a critical treatment strategy for irreversible intestinal failure. Among all abdominal solid organ transplants, the intestine was the most vulnerable to ischemia and reperfusion injury (IRI). The static cold storage (SCS) technique is currently the most commonly used graft preservation method, but its hypoxia condition causes metabolic disorders, resulting in the occurrence of IRI, limiting its application in marginal organs. It is especially important to improve preservation techniques in order to minimize damage to marginal donor organs, which draws more attention to machine perfusion (MP). There has been much debate about whether it is necessary to increase oxygen in these conditions to support low levels of metabolism since the use of machine perfusion to preserve organs. There is evidence that oxygenation helps to restore intracellular ATP levels in the intestine after thermal or cold ischemia damage. The goal of this review is to provide an overview of the role of oxygen in maintaining environmental stability in the gut under hypoxic conditions, as well as to investigate the possibilities and mechanisms of oxygen delivery during preservation in intestine transplantation studies and clinical models.


A positive feedback circuit driven by m6A-modified circular RNA facilitates colorectal cancer liver metastasis.
Liver metastasis is the leading cause of death in patients with colorectal cancer (CRC). Emerge evidence suggests that circular RNA (circRNA) is a pivotal player in cancer progression. However, its role in CRC liver metastasis remains largely unknown. Circ-YAP expression was detected by qRT-PCR and in situ hybridization. The function of circ-YAP was tested by wound healing, transwell and CCK-8 assays. RNA immunoprecipitation, pull-down, luciferase reporter, chromatin immunoprecipitation assays were used to investigate the mechanism underlying circ-YAP promoting CRC liver metastasis. CRC liver metastasis animal model was established to assess the effect of circ-YAP in vivo. Circ-YAP was notably upregulated in CRC with liver metastasis, which was associated with dismal prognosis. Circ-YAP promoted CRC cell migration and invasion in vitro, and facilitated liver metastasis in patient-derived xenografts (PDX) models in vivo. Mechanistically, circ-YAP encoded a novel truncated protein containing 220 amino acids, termed as YAP-220aa, which competitively bound to LATS1, resulting in YAP dephosphorylation and nuclear translocation, thereby activating a cohort of metastasis-promoting genes. Importantly, N6-methyladenosine (m6A) modification orchestrated efficient initiation of circ-YAP translation, requiring m6A reader YTHDF3 and eIF4G2 translation initiation complex. Intriguingly, circ-YAP was transcriptionally enhanced by YAP/TEAD complex, thus forming a positive regulatory feed-forward loop. Our findings reveal a previously uncharacterized oncoprotein encoded by circ-YAP, implying a promising biomarker and therapeutic target for CRC patients with liver metastasis.


Extreme hyperphosphatemia and acute renal failure after a phosphorus-containing bowel regimen.
Phosphate intoxication, manifested by hypocalcemic tetany and acute renal failure, may complicate bowel-cleansing preparations which contain phosphate. These preparations are commonly used to prepare patients for various gastrointestinal procedures. Often, patients who receive these regimens are at increased risk of phosphate intoxication from diseases which slow gastrointestinal transit or decrease renal excretion (renal insufficiency). We present a patient who developed oliguric acute renal failure from severe phosphate intoxication associated with a phosphate-containing bowel-cleansing regimen.


Uridine monophosphate kinase as potential target for tuberculosis: from target to lead identification.
Mycobacterium tuberculosis (Mtb) is a causative agent of tuberculosis (TB) disease, which has affected approximately 2 billion people worldwide. Due to the emergence of resistance towards the existing drugs, discovery of new anti-TB drugs is an important global healthcare challenge. To address this problem, there is an urgent need to identify new drug targets in Mtb. In the present study, the subtractive genomics approach has been employed for the identification of new drug targets against TB. Screening the Mtb proteome using the Database of Essential Genes (DEG) and human proteome resulted in the identification of 60 key proteins which have no eukaryotic counterparts. Critical analysis of these proteins using Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathways database revealed uridine monophosphate kinase (UMPK) enzyme as a potential drug target for developing novel anti-TB drugs. Homology model of Mtb-UMPK was constructed for the first time on the basis of the crystal structure of E. coli-UMPK, in order to understand its structure-function relationships, and which would in turn facilitate to perform structure-based inhibitor design. Furthermore, the structural similarity search was carried out using physiological inhibitor UTP of Mtb-UMPK to virtually screen ZINC database. Retrieved hits were further screened by implementing several filters like ADME and toxicity followed by molecular docking. Finally, on the basis of the Glide docking score and the mode of binding, 6 putative leads were identified as inhibitors of this enzyme which can potentially emerge as future drugs for the treatment of TB.


Role of a behavioral therapy involving psychodrama for the treatment of sleep disorders in pediatric primary care.
Sleep disorders are common in young children. Our objective was to describe a psychodrama using puppets and to assess the interest of this approach for the treatment of sleep disorders in ambulatory pediatric patients. This retrospective, observational, monocenter study was carried between 1st January 2014 and 31st December 2015. Children aged 12 months to 6 years with a sleep disorder confirmed according to the International Classification of Sleep Disorders-2 were recruited. The parents were questioned separately about their child's sleep rhythm and the family's quality of life (QoL). The child sitting on their parent's knee and, using puppets to represent the family members, the pediatrician reproduced the scenario that took place at home and demonstrated what should be changed. The parents were contacted by telephone 1-2 weeks after the therapy to determine whether the child's sleep patterns had improved. The primary endpoint was resolution of the sleep disorder. Thirty-eight children (mean age 27.2±14.0 months; mean duration of sleep disorder 12.7±9.5 months) were assessed. The most common sleep disorders were difficulties in falling asleep at evening bed-time (76.3%) and night-time wakening (76.3%). The main triggering factors were the birth of a sibling (30.8%) or an illness (30.8%). In the majority (52.2%) of families who were convinced that the psychodrama would work, an immediate and complete resolution of the sleep disorder was obtained within three days of the consultation. QoL improved in the majority of families after the consultation (QoL was average in 68.4% of families before the consultation vs. good in 84.2% after the consultation). Psychodrama was effective at bringing about an immediate and complete resolution of sleep disorders in children. This confirms the findings of other reports that show that behavioral therapy is effective in this context. This method could be adapted effectively by individual practitioners, enabling children to establish a good sleep pattern without the use of medications. A further large-scale, randomized, controlled trial is required to confirm these results.


Incidence and determinants of hyperkalemia and hypokalemia in a large healthcare system.
Hypo- and hyperkalemia in clinical settings are insufficiently characterized and large-scale data from Europe lacking. We studied incidence and determinants of these abnormalities in a large Swedish healthcare system. Observational study from the Stockholm CREAtinine Measurements project, including adult individuals from Stockholm accessing healthcare in 2009 (n=364,955). Over 3-years, we estimated the incidence of hypokalemia, defined as potassium<3.5mmol/L, hyperkalemia, defined as potassium>5mmol/L, and moderate/severe hyperkalemia, defined as potassium>5.5mmol/L. Kidney function was assessed by estimated glomerular filtration rate (eGFR). Of 364,955 participants, 69.4% had 1-2 potassium tests, 16.7% had 3-4 tests and the remaining 13.9% had >4potassiumtests/year. Hypokalemia occurred in 49,662 (13.6%) individuals, with 33% recurrence. Hyperkalemia occurred in 25,461 (7%) individuals, with 35.7% recurrence. Moderate/severe hyperkalemia occurred in 9059 (2.5%) with 28% recurrence. The frequency of potassium testing was an important determinant of dyskalemia risk. The incidence proportion of hyperkalemia was higher in the presence of diabetes, lower eGFR, myocardial infarction, heart failure (HF), or use of renin angiotensin-aldosterone system inhibitors (RAASi). In adjusted analyses, women and use of loop/thiazide diuretics were associated with lower hyperkalemia risk. Older age, lower eGFR, diabetes, HF and use of RAASi were associated with higher hyperkalemia risk. On the other hand, women, younger age, higher eGFR and baseline use of diuretics were associated with higher hypokalemia risk. Hypo- and hyperkalemia are common in healthcare. Optimal RAASi and diuretics use and careful potassium monitoring in the presence of certain comorbidities, especially lower eGFR, is advocated.


BAMM gives misleading rate estimates in simulated and empirical datasets.
In a previous paper, we used simulations and empirical data to show that BAMM (Bayesian Analysis of Macroevolutionary Mixtures) can give misleading estimates of rates and rate shifts. In simulations, BAMM underestimated rate shifts across every tree analyzed, and assigned incorrect rates to most clades in most trees. In empirical analyses, BAMM behaved as expected from simulations, and assigned different rates to clades when clades were analyzed alone versus across the tree (i.e., with rate heterogeneity). Rabosky recently criticized our paper, focusing primarily on the idea that our comparison of BAMM to another approach (method-of-moments estimators of Magallón and Sanderson, or MS estimators) was unfair to BAMM. Here, we provide further evidence that BAMM gives misleading rate estimates in empirical studies. We then describe how Rabosky's rown method comparisons were either acknowledged as being problematic or were described inaccurately (to favor BAMM). Finally, we show that the MS estimators can perform well when rates vary over time, despite untested assertions that they require constant rates to be accurate. Many other methods are available for analyzing diversification rates: we argue that BAMM should be avoided for estimating both diversification rates and rate shifts.


Microphotometric differentiation of human T and B cells tagged with monospecific immunoadsorbent beads.
A comparative and statistical study was done of the classification of peripheral blood lymphocytes (PBLs) as T cells, B cells and monocytes by various immunologic procedures and computerized microphotometric analysis. PBLs from 15 healthy male subjects were examined by immunofluorescence, E rosettes, and immunoadsorbent beads (IAB) for T cells and B cells, by phase contrast microscopy and as fixed slide preparations. Cells tagged with IAB for T cells and B cells were fixed, stained with Papanicolaou stain and analyzed. Evaluation of immunologic data shows 50% to 57% T cells, 9.7% to 24.1% B cells, 13.4% to 16.1% monocytes and about 20% unmarked cells. Analysis of T cells shows significant correlations between E and T cell-IAB rosettes, but neither rosetting procedure reveals a positive correlation with immunofluorescence-labelled T cells. Comparison of B cells shows an insignificant correlation between B-cell IAB and immunofluorescence. Results showed that permanent slides of rosetted cells can be made without alteration in relative numbers of rosetted cells. Assessment of immunologically tagged cell samples by image analysis correctly classified 80% to 90% of cells as T and B cells. Evaluation of homogeneity of the T and B cell populations shows the existence of four subsets of B cells and five subsets of T cells.


Anterior innominate osteotomy in repair of bladder exstrophy.
Classic bladder exstrophy is a developmental defect presenting at birth with a wide pubic separation and an exposed bladder; cloacal exstrophy involves, in addition, intestinal prolapse. Reconstruction requires several surgical procedures. The use of anterior iliac osteotomies in this process has not been reviewed in a large series. We reviewed the results of eighty-six anterior innominate osteotomies performed in conjunction with genitourinary repair of classic and cloacal bladder exstrophy in eighty-two patients. Clinical outcome measures were successful bladder closure, achievement of continence, and maintenance of a normal gait. Radiographs of the pelvis were reviewed, and the pubic intersymphyseal diastasis (a measure of the reduction in tension on the anterior closure) was measured preoperatively and at three time-points postoperatively. Children with classic exstrophy who had undergone osteotomy and bladder neck reconstruction but not bladder augmentation were divided into four groups on the basis of the degree of continence. In addition, children with classic exstrophy were stratified according to age at the time of the osteotomy. The mean postoperative percent reduction in the amount of the original diastasis was determined for all groups. Children with classic exstrophy and those with cloacal exstrophy had correction of the diastasis after the osteotomy, with greater correction in those with classic exstrophy, presumably because of better bone quality. Daytime continence was achieved with anterior osteotomy and bladder neck reconstruction in 74% of the children for whom continence was a goal. However, no difference in the symphyseal diastasis or in the percentage of pubic reduction was detected among the four continence groups. Children who were older at the time of the osteotomy maintained better correction over time. Wound dehiscence or bladder prolapse occurred in 4% of the patients who had osteotomy and primary closure, and the only important complication of the osteotomies was transient palsy of the left femoral nerve in seven children. Anterior innominate osteotomy is an effective part of reconstructive repair of bladder exstrophy. The primary goals of the osteotomy are to reduce the tension in the closed bladder and the lower abdominal wall and to promote continence by restoring the sling of the pelvic floor muscles. These goals can be achieved in the majority of patients.


Suicide and 'violent' death in a six-year cohort of male probationers compared with pattern of mortality in the general population: evidence of accumulative socio-psychiatric vulnerability.
The 'Health of the Nation' (Department of Health, 1992) suicide targets focus upon the mentally ill, but virtually ignore the mentally abnormal offender. Whilst forensic services deal with the severely disturbed, the majority of offenders remain in the community, despite long-standing psychosocial difficulties. This study explores the mortality rates of a six-year cohort of male probationers (1990-1995) with males in the general population. Male offenders (aged 17-54) had double the death rate, five times the 'external death' rate and nine times the suicide rate of the general population. This paper highlights the need to further improve the health-psychiatric-criminal justice collaboration.


Spontaneous Subcapsular Renal Haematoma : A Case Report.
Spontaneous subcapsular renal haematoma is a rare condition with wide range of presentation; it poses a dilemma for diagnosis and management. We present a case of 38-year-old female who presented with right flank pain (continuous, dull aching) for a week with right renal angle tenderness and high blood pressure (though she was not known hypertensive). On imaging, there was right renal subcapsular collection. She did not respond to conservative management except that her blood pressure was controlled with single drug Angiotensin Converting Enzyme (ACE) inhibitor. Right double J stent was placed (in view of urinoma) and patient was followed for six weeks. Repeat computed tomography scan showed persistence of right renal subcapsular collection but the cause was not found. Except hypertension, no definitive cause for the condition could be found. Patient was intervened surgically with right subcostal exploration and subcapsular haematoma was found and drained. Patient was asymptomatic thereafter.


Proteomics of human umbilical vein endothelial cells applied to etoposide-induced apoptosis.
We have undertaken to continue the proteomic study of human umbilical vein endothelial cells (HUVECs) using the combination of 2-DE, automated trypsin digestion, and PMF analysis after MALDI-TOF MS and peptide sequencing using nano LC-ESI-MS/MS. The overall functional characterization of the 162 identified proteins from primary cultures of HUVECs confirms the metabolic capabilities of endothelium and illustrates various cellular functions more related to cell motility and angiogenesis, protein folding, anti-oxidant defenses, signal transduction, proteasome pathway and resistance to apoptosis. In comparison with controls cells, the differential proteomic analysis of HUVECs treated by the pro-apoptotic topoisomerase inhibitor etoposide further revealed the variation of eight proteins, namely, GRP78, GRP94, valosin-containing protein, proteinase inhibitor 9, cofilin, 37-kDa laminin receptor protein, bovine apolipoprotein, and tropomyosin. These data suggest that etoposide-induced apoptosis of human vascular endothelial cells results from the intricate involvement of multiple apoptosis processes including at least the mitochondrial and the ER stress pathways. The presented 2-D pattern and protein database, as well as the data related to apoptosis of HUVECs, are available at http://www.huvec.com.


Utility of diagnostic ultrasound in evaluating fracture healing.
With increase in population, modes of transportation and a fast pace of life, an individual's chances of accident and thereby chances of getting fractured have increased significantly. Fracture has thus become a significant factor contributing to morbidity and mortality. To resume a normal life, after one suffers from a fracture is also an ordeal. The transition appears smooth if the fracture healing goes on in a smooth manner as is expected through its routine stages of reactive phase, reparative phase and remodeling phase. But if in this chain something goes wrong or some factors are not optimum upto the mark, then the process becomes unsuccessful and the repair is either partial or directionless. It is therefore very vital to confirm whether or not the callus which bridges the fractured fragments is healthy or not. Here in lies the role of imaging as it can show the status of callus without disturbing it. What complicates the picture is that a callus might not be well demonstratable unless it mineralizes or calcifies. An imaging modality like ultrasound therefore stands out as it can show the state of callus in its different stages. This article aims at demonstrating how ultrasound- a non invasive diagnostic imaging modality can give precise information about the progress of fracture healing and thereby aid in management of fractures, so that an individual can return back to normal productive lifestyle. This preliminary study highlights the spectrum of fracture healing as seen on ultrasound.


Does the Fast Track Intervention Prevent Later Psychosis Symptoms?
The Fast Track (FT) intervention was a multimodal preventive intervention addressing antisocial development across 10 years of childhood and early adolescence. The intervention included parent management training, child social-cognitive skills training, peer coaching and mentoring, academic skills tutoring, and a classroom social-emotional learning program. While not specifically designed to target psychosis symptoms (e.g., social withdrawal, thought abnormalities), the present study aimed to examine whether the FT intervention prevented psychosis symptoms through childhood and adolescence and into adulthood. Participants included the FT intervention and high-risk control samples (N = 891; 69% male; M age = 6.58 years, SD = .48). Psychosis symptoms were assessed using the "thought problems" subscale of the parent-report Child Behavior Checklist during grades 1, 2, 4, 5, and 7, and the self-report Adult Behavior Checklist at age 25 years, in line with prior research using this measure. Growth models included the FT condition and covariates (i.e., initial risk screen score, cohort, socioeconomic status, rural/urban status, race, and sex) as predictors; and child, adolescent, and adult psychosis symptoms as outcomes. Intervention status was not significantly associated with the slope of psychosis symptoms; however, after controlling for concurrent cannabis use, intervention participants reported lower levels of psychosis symptoms over time. Findings suggest that interventions targeting antisocial behavior may prevent psychosis symptoms in the long term.


Dissociation of second messenger activation by parathyroid hormone fragments in osteosarcoma cells.
PTH activates multiple second messengers in its target cells, but the level at which the hormonal signal splits into different pathways is still unknown. To achieve insights on this issue, we have studied the structure-function relationship of PTH by analyzing the effects of bovine PTH-(1-34) [bPTH-(1-34)] and PTH fragments truncated at the N-terminus on the intracellular calcium concentration [( Ca2+]i) and cAMP production in the rat osteogenic sarcoma cell line UMR 106-01. [Ca2+]i was measured in single cells using fura-2. When exposed to 10(-7) M bPTH-(1-34), 20% of the cells responded with a transient increase in [Ca2+]i of variable amplitude. Equimolar doses of bPTH-(2-34), propionyl bPTH-(2-34) [(pbPTH-(2-34)], and bPTH-(3-34) also transiently increased [Ca2+]i, whereas both [tyrosine34]bPTH-(7-34) amide [bPTH-(7-34)] and bPTH-(30-34) were ineffective. The amplitude of the [Ca2+] i transients was dose-dependent, with threshold concentrations of 10(-10) M for bPTH-(1-34) and bPTH-(2-34), and 10(-9) M for bPTH-(3-34). The response rate to the active peptides ranged between 10-30%, without a clear dose-relatedness. A second addition of 10(-7) M bPTH-(1-34) to cells prestimulated with equimolar doses of bPTH-(2-34), pbPTH-(2-34), or bPTH-(3-34) produced another transient, whereas after exposure to 10(-7) M bPTH-(1-34), the cells were completely desensitized to a second homologous stimulation, suggesting that the binding affinity of the truncated peptides for the PTH receptor is lower than that of the intact bPTH-(1-34) fragment. In addition, both bPTH-(1-34) and bPTH-(2-34) dose-dependently stimulated cAMP production, but the former was more potent (ED50 = 10(-9) vs. 10(-7) M, respectively). On the contrary, pbPTH-(2-34), bPTH-(3-34), and bPTH-(7-34) had no effect on cAMP. Pretreating the cells with pertussis toxin to enhance cAMP responses via inhibition of Gi potentiated the effect of bPTH-(1-34) and bPTH-(2-34) and disclosed weak but detectable agonist action of pbPTH-(2-34). These results indicate that specific domains of the PTH molecule are linked to activation of different second messenger pathways; while the first two amino acids are indispensable for activating the cAMP system, generation of the [Ca2+]i signal appears to involve a longer domain, including the amino acid residue in position 3.


Perchloro-2,5,8-triazaphenalenyl radical.
[reaction: see text] The unusually stable perchloro-2,5,8-triazaphenalenyl radical 1 and its twisted dechlorinated dimer 2 were synthesized and characterized by ESR spectroscopy and X-ray crystallography. The X-ray structure of dimer 2 shows that the double bond connecting the two triazaphenalene systems is strongly twisted. Dimer 2 has a dramatic color shift from the solid state to solution, which may be due to a change of the twisting angle between both states.


A stop or go switch: glycogen synthase kinase 3β phosphorylation of the kinesin 1 motor domain at Ser314 halts motility without detaching from microtubules.
It is more than 25 years since the discovery that kinesin 1 is phosphorylated by several protein kinases. However, fundamental questions still remain as to how specific protein kinase(s) contribute to particular motor functions under physiological conditions. Because, within an whole organism, kinase cascades display considerable crosstalk and play multiple roles in cell homeostasis, deciphering which kinase(s) is/are involved in a particular process has been challenging. Previously, we found that GSK3β plays a role in motor function. Here, we report that a particular site on kinesin 1 motor domain (KHC), S314, is phosphorylated by GSK3β in vivo. The GSK3β-phosphomimetic-KHCS314D stalled kinesin 1 motility without dissociating from microtubules, indicating that constitutive GSK3β phosphorylation of the motor domain acts as a STOP. In contrast, uncoordinated mitochondrial motility was observed in CRISPR/Cas9-GSK3β non-phosphorylatable-KHCS314A Drosophila larval axons, owing to decreased kinesin 1 attachment to microtubules and/or membranes, and reduced ATPase activity. Together, we propose that GSK3β phosphorylation fine-tunes kinesin 1 movement in vivo via differential phosphorylation, unraveling the complex in vivo regulatory mechanisms that exist during axonal motility of cargos attached to multiple kinesin 1 and dynein motors.


Increased pain and muscle glutamate concentration after single ingestion of monosodium glutamate by myofascial temporomandibular disorders patients.
A randomized, double-blinded, placebo-controlled study was conducted to investigate if single monosodium glutamate (MSG) administration would elevate muscle/serum glutamate concentrations and affect muscle pain sensitivity in myofascial temporomandibular disorders (TMD) patients more than in healthy individuals. Twelve myofascial TMD patients and 12 sex- and age-matched healthy controls participated in two sessions. Participants drank MSG (150 mg/kg) or NaCl (24 mg/kg; control) diluted in 400 mL of soda. The concentration of glutamate in the masseter muscle, blood plasma and saliva was determined before and after the ingestion of MSG or control. At baseline and every 15 min after the ingestion, pain intensity was scored on a 0-10 numeric rating scale. Pressure pain threshold, pressure pain tolerance (PPTol) and autonomic parameters were measured. All participants were asked to report adverse effects after the ingestion. In TMD, interstitial glutamate concentration was significantly greater after the MSG ingestion when compared with healthy controls. TMD reported a mean pain intensity of 2.8/10 at baseline, which significantly increased by 40% 30 min post MSG ingestion. At baseline, TMD showed lower PPTols in the masseter and trapezius, and higher diastolic blood pressure and heart rate than healthy controls. The MSG ingestion resulted in reports of headache by half of the TMD and healthy controls, respectively. These findings suggest that myofascial TMD patients may be particularly sensitive to the effects of ingested MSG. WHAT DOES THIS STUDY ADD?': Elevation of interstitial glutamate concentration in the masseter muscle caused by monosodium glutamate (MSG) ingestion was significantly greater in myofascial myofascial temporomandibular disorders (TMD) patients than healthy individuals. This elevation of interstitial glutamate concentration in the masseter muscle significantly increased the intensity of spontaneous pain in myofascial TMD patients.


Aberrant splicing of tau pre-mRNA caused by intronic mutations associated with the inherited dementia frontotemporal dementia with parkinsonism linked to chromosome 17.
Frontotemporal dementia accounts for a significant fraction of dementia cases. Frontotemporal dementia with parkinsonism linked to chromosome 17 is associated with either exonic or intronic mutations in the tau gene. This highlights the involvement of aberrant pre-mRNA splicing in the pathogenesis of neurodegenerative disorders. Little is known about the molecular mechanisms of the splicing defects underlying these diseases. To establish a model system for studying the role of pre-mRNA splicing in neurodegenerative diseases, we have constructed a tau minigene that reproduces tau alternative splicing in both cultured cells and in vitro biochemical assays. We demonstrate that mutations in a nonconserved intronic region of the human tau gene lead to increased splicing between exon 10 and exon 11. Systematic biochemical analyses indicate the importance of U1 snRNP and, to a lesser extent, U6 snRNP in differentially recognizing wild-type versus intron mutant tau pre-mRNAs. Gel mobility shift assays with purified U1 snRNP and oligonucleotide-directed RNase H cleavage experiments support the idea that the intronic mutations destabilize a stem-loop structure that sequesters the 5' splice site downstream of exon 10 in tau pre-mRNA, leading to increases in U1 snRNP binding and in splicing between exon 10 and exon 11. Thus, mutations in nonconserved intronic regions that increase rather than decrease alternative splicing can be an important pathogenic mechanism for the development of human diseases.


Aging affects the retinoic acid and the triiodothyronine nuclear receptor mRNA expression in human peripheral blood mononuclear cells.
Inadequate retinoid status has often been described as occurring with aging. Moreover, subclinical hypothyroid status has also been evoked in the elderly. Several studies performed in animals have described the crucial incidence of age-related hypo-functioning of retinoid and thyroid signalling pathways, particularly in the brain. The aim of the present study was to clarify whether aging modifies retinoid and thyroid signalling in humans. Using real-time RT-PCR the relative amount of mRNA of the retinoid (RARalpha, RARgamma and RXRalpha) and thyroid (TRalpha and TRbeta) nuclear receptors in peripheral blood mononuclear cells (PBMC) of young (24-57 years old, n = 22) compared with elderly (69-90 years old, n = 24) healthy subjects was quantitated. Classical plasma parameters used to characterize the retinoid and thyroid status - retinol (ROH), retinol-binding protein (RBP), free triiodothyronine (FT3) and thyroxine (FT4), thyroid-stimulating hormone (TSH) and transthyretin (TTR) - were also assessed. RARgamma expression was significantly decreased in elderly versus young subjects while no modification of the retinoid-related plasma parameters ROH and RBP were emphasized by aging. Concerning thyroid criteria, the elderly exhibited an increase in TSH concentration (+39%) without significant modifications of FT3 and FT4, which indicated an age-related sub-clinical hypothyroidism. Concurrently, the amount of TR mRNA (alpha as well as beta subtypes) was significantly decreased in the elderly. These data constitute the first evidence of an age-related hypo-activation of the retinoid and thyroid nuclear pathways in PBMC. Further study of the possible association between the expression of the retinoid and thyroid nuclear receptors and age-related cognitive alterations in humans would be interesting.


[Relationship of DNMT3b expression and SEPT9 methylation in theprogression of colorectal carcinogenesis].
Objective: To investigate the relationship between the expression of DNA methyltransferase 3b (DNMT3b) and the methylation of SEPT9 gene, and their application prospects in the diagnosis and treatment of colorectal cancer. Methods: Seventy-five cases of colorectal cancer and adjacent tissues, 68 cases of colorectal high-grade internal neoplasia tissues (referred to as precancerous tissues) and high-grade internal adjacent neoplasia tissues (referred to as adjacent precancerous tissues) were collected. Pyrosequencing was used to detect the methylationlevel of SETP9. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to examine the mRNA expressionof SEPT9 and immunohistochemistry(IHC) was applied to detect the protein expressions of SETP9 and DNMT3b. Liposome-mediated method was used to transfect DNMT3b siRNA and negative control siRNA into HT-29 cells. Five groups including DNMT3b siRNA 15 nmol/L group, DNMT3b siRNA 30 nmol/L group, negative control siRNA 15 nmol/L group, negative control siRNA 30 nmol/L group and blank control group were set up. Pyrosequencing was applied to determine the methylation level of SEPT9 and mRNA expression of DNMT3b in each group. Results: The methylation rates of SEPT9 gene in colorectal cancer tissues, adjacent tissues, precancerous tissues and adjacent precancerous tissues were (76.8±6.5)%, (14.4±2.6)%, (34.6±5.0)% and (7.4±1.2)%, respectively, which was highest in colorectal cancer tissue (P<0.001). The relative expressions of SEPT9 mRNA were 0.18±0.03, 0.89±0.41, 0.69±0.41 and 1.01±0.21, respectively, which was lowest in colorectal cancer tissue (P<0.001), while there were no statistically significant differences in adjacent tissues, precancerous tissues and adjacent precancerous tissues (P>0.05). The positive rates of SEPT9 protein expression were 12.0% (9/75), 53.3% (40/75), 55.1% (38/69) and 62.3% (43/69), which was lowest in the colorectal cancer tissue (P<0.001), while there were no statistically significant differences in the adjacent group, precancerous group and adjacent precancerous group (P>0.016 7). The positive rates of DNMT3b protein expression were 56.3% (45/75), 26.7% (20/75), 46.4% (32/69) and 33.3% (23/69), respectively, which was highest in colorectal cancer tissue (P<0.001), while without statistically significant difference from the precancerous tissue (P>0.016 7). Experiments in vitro showed that DNMT3b mRNA expression was lowest in DNMT3b siRNA 30 nmol/L group among five groups and was statistically different from other groups (all P<0.05). Meanwhile, the methylationrate of SEPT9 gene was lowest in this group, but without statistically significant difference from the DNMT3b siRNA 15 nmol/L group (P>0.05). Conclusions: The expression of DNMT3b is significantly correlated with the methylation level of SEPT9 gene in different stages of colorectal cancer. The high expression of DNMT3b may be an important molecular event before SEPT9 gene methylation and it may have an important potential application value in the diagnosis and treatment of early colorectal cancer.


Functional and cortical adaptations to central vision loss.
Age-related macular degeneration (AMD), affecting the retina, afflicts one out of ten people aged 80 years or older in the United States. AMD often results in vision loss to the central 15-20 deg of the visual field (i.e. central scotoma), and frequently afflicts both eyes. In most cases, when the central scotoma includes the fovea, patients will adopt an eccentric preferred retinal locus (PRL) for fixation. The onset of a central scotoma results in the absence of retinal inputs to corresponding regions of retinotopically mapped visual cortex. Animal studies have shown evidence for reorganization in adult mammals for such cortical areas following experimentally induced central scotomata. However, it is still unknown whether reorganization occurs in primary visual cortex (V1) of AMD patients. Nor is it known whether the adoption of a PRL corresponds to changes to the retinotopic mapping of V1. Two recent advances hold out the promise for addressing these issues and for contributing to the rehabilitation of AMD patients: improved methods for assessing visual function across the fields of AMD patients using the scanning laser ophthalmoscope, and the advent of brain-imaging methods for studying retinotopic mapping in humans. For the most part, specialists in these two areas come from different disciplines and communities, with few opportunities to interact. The purpose of this review is to summarize key findings on both the clinical and neuroscience issues related to questions about visual adaptation in AMD patients.


[Post traumatic myositis ossificans of the iliopsoas muscle. Apropos of a case with review of the literature].
A case of traumatic myositis ossificans of the iliopsoas muscle in a 13 year old ballet dancer is presented. A history of multiple minor injuries to the region of the left hip in addition to the radiographic appearance of calcification were indicative of ectopic bone formation. The lesion was explored surgically because of the clinical symptoms of the coexistant femoral hernia as well as to obtain biopsy material to exclude a soft tissue tumor. Histological examination demonstrated the zoning effect of myositis ossificans with patterns varying from a cellular central zone to a peripheral one of fairly well oriented bone. The patient remains symptom-free and no radiographic evidence of recurrence was observed four years postoperatively.


A conserved RpoS-dependent small RNA controls the synthesis of major porin OmpD.
A remarkable feature of many small non-coding RNAs (sRNAs) of Escherichia coli and Salmonella is their accumulation in the stationary phase of bacterial growth. Several stress response regulators and sigma factors have been reported to direct the transcription of stationary phase-specific sRNAs, but a widely conserved sRNA gene that is controlled by the major stationary phase and stress sigma factor, σ(S) (RpoS), has remained elusive. We have studied in Salmonella the conserved SdsR sRNA, previously known as RyeB, one of the most abundant stationary phase-specific sRNAs in E. coli. Alignments of the sdsR promoter region and genetic analysis strongly suggest that this sRNA gene is selectively transcribed by σ(S). We show that SdsR down-regulates the synthesis of the major Salmonella porin OmpD by Hfq-dependent base pairing; SdsR thus represents the fourth sRNA to regulate this major outer membrane porin. Similar to the InvR, MicC and RybB sRNAs, SdsR recognizes the ompD mRNA in the coding sequence, suggesting that this mRNA may be primarily targeted downstream of the start codon. The SdsR-binding site in ompD was localized by 3'-RACE, an experimental approach that promises to be of use in predicting other sRNA-target interactions in bacteria.


Species' ecological functionality alters the outcome of fish stocking success predicted by a food-web model.
Fish stocking is used worldwide in conservation and management, but its effects on food-web dynamics and ecosystem stability are poorly known. To better understand these effects and predict the outcomes of stocking, we used an empirically validated network model of a well-studied lake ecosystem. We simulate two stocking scenarios with two native fish species valuable for fishing. In the first scenario, we stock planktivorous fish (whitefish) larvae in the ecosystem. This leads to a 1% increase in adult whitefish biomasses and decreases the biomasses of the top predator (perch). In the second scenario, we also stock perch larvae in the ecosystem. This decreases the planktivorous whitefish and the oldest top predator age class biomasses, and destabilizes the ecosystem. Our results demonstrate that the effects of stocking depend on the species' position in the food web and thus cannot be assessed without considering interacting species. We further show that stocking can lead to undesired outcomes from both management and conservation perspectives. The gains of stocking can remain minor and have adverse effects on the entire ecosystem.


Multinational, double-blind, randomised, placebo-controlled, prospective study of esomeprazole in the prevention of recurrent peptic ulcer in low-dose acetylsalicylic acid users: the LAVENDER study.
To evaluate if esomeprazole prevents recurrent peptic ulcer in adult patients with a history of peptic ulcer receiving low-dose acetylsalicylic acid (ASA, aspirin) for cardiovascular protection in East Asia. In this prospective, randomised, double-blind, placebo-controlled trial conducted in Japan, Korea and Taiwan, eligible patients receiving low-dose ASA for cardiovascular protection (81-324 mg/day) were randomised to esomeprazole 20 mg/day or placebo for ≤72 weeks. All patients received concomitant mucosal protection (gefarnate 100 mg/day). The primary endpoint was time to ulcer recurrence (Kaplan-Meier analysis). Efficacy findings are presented up to week 48, as per a planned interim analysis within the study protocol. A total of 364 patients (79.9% men; mean age, 67.1 years) comprised the full analysis set (esomeprazole, n=182; placebo, n=182). There was a statistically significant difference in the time to ulcer recurrence between esomeprazole and placebo (HR 0.09; 96.65% CI 0.02 to 0.41; p<0.001). The estimated ulcer-free rate at week 12 was 99.3% (esomeprazole) and 89.0% (placebo). The high estimated ulcer-free rate for esomeprazole was maintained through to week 48 (98.3% vs. 81.2% of placebo-treated patients). No factors, other than female gender, reduced time to ulcer recurrence in addition to the effect of esomeprazole (p<0.001). Treatment with esomeprazole was generally well tolerated. Daily esomeprazole 20 mg is efficacious and well tolerated in reducing the recurrence of peptic ulcer in East-Asian patients with a history of ulcers who are taking low-dose ASA for cardiovascular protection. NCT01069939.


[Effectiveness of bemitil in recurrent erysipelas].
The trial entered 66 patients with recurrent erysipelas treated conventionally with addition of either immunostimulator bemitil (0.25-0.5 g/day orally for 5-7 days) or placebo. The bemitil group was free of intoxication symptoms and local manifestations, discharged from hospital sooner than the controls. Therapeutic efficacy of bemitil is due to its promotion of mononuclear phagocytes function which acts as an essential mechanism of antibacterial defence activation in patients with recurrent erysipelas.


Watching in situ the hydrogen diffusion dynamics in magnesium on the nanoscale.
Active plasmonic and nanophotonic systems require switchable materials with extreme material contrast, short switching times, and negligible degradation. On the quest for these supreme properties, an in-depth understanding of the nanoscopic processes is essential. Here, we unravel the nanoscopic details of the phase transition dynamics of metallic magnesium (Mg) to dielectric magnesium hydride (MgH2) using free-standing films for in situ nanoimaging. A characteristic MgH2 phonon resonance is used to achieve unprecedented chemical specificity between the material states. Our results reveal that the hydride phase nucleates at grain boundaries, from where the hydrogenation progresses into the adjoining nanocrystallites. We measure a much faster nanoscopic hydride phase propagation in comparison to the macroscopic propagation dynamics. Our innovative method offers an engineering strategy to overcome the hitherto limited diffusion coefficients and has substantial impact on the further design, development, and analysis of switchable phase transition as well as hydrogen storage and generation materials.


Characterization of mosquito breeding sites in and in the vicinity of tigray microdams.
Malaria vector control in Ethiopia has a history of more than 50 years, but malaria remains a major cause of morbidity and mortality in Ethiopia. Thus, targeting the control program on the larval stage is of paramount importance. This study aimed to characterize the aquatic habitats of vector mosquito larvae associated with micro-dams. Cross-sectional larval survey was conducted on six micro-dams in Tigray, Northern Ethiopia in 2005/06. The study area on each dam was divided into eight zones. Immature stages of mosquitoes were collected using standard dippers. The physico-chemical characteristics of the aquatic habitats were measured onsite. A total of 301 aquatic habitats were surveyed for mosquito larvae; in 32.56% (n=301) only Anopheles, in 27.91% only Culex, both genera were found mixed in 21.59% and no mosquito larvae were found in 17.94%. The findings depicted that dissolved oxygen (r = 0.34, p =0.04), pH (r = 0.35, p =0.03), conductivity (r = 0.36, p =0.03), vegetation (F = 3.54, p =0.002), microhabitat (F = 2.65, p =0.04), fauna and bottom surface of the water body were positively associated and important in explaining the presence and abundance of Culex. On the other hand, dissolved oxygen (r = 0.39, p =0.02), pH (r = 0.42, p =0.008), vegetation (F = 5.6, p =0.000), water transparency (F = 2.72, p =0.00), rainfall (F = 2.22, p = 0.027) and fauna were positively associated and important in explaining the presence and abundance of Anopheles. The findings of this study suggest that both biotic (vegetation and fauna) and abiotic (chemical and physical) factors play a significant role in larvae's habitat preference in both Culex and Anopheles.


Close, stable homolog juxtaposition during meiosis in budding yeast is dependent on meiotic recombination, occurs independently of synapsis, and is distinct from DSB-independent pairing contacts.
A site-specific recombination system that probes the relative probabilities that pairs of chromosomal loci collide with one another in living cells of budding yeast was used to explore the relative contributions of pairing, recombination, synaptonemal complex formation, and telomere clustering to the close juxtaposition of homologous chromosome pairs during meiosis. The level of Cre-mediated recombination between a pair of loxP sites located at an allelic position on homologous chromosomes was 13-fold greater than that between a pair of loxP sites located at ectopic positions on nonhomologous chromosomes. Mutations affecting meiotic recombination initiation and the processing of DNA double-strand breaks (DSBs) into single-end invasions (SEIs) reduced the levels of allelic Cre-mediated recombination levels by three- to sixfold. The severity of Cre/loxP phenotypes is presented in contrast to relatively weak DSB-independent pairing defects as assayed using fluorescence in situ hybridization for these mutants. Mutations affecting synaptonemal complex (SC) formation or crossover control gave wild-type levels of allelic Cre-mediated recombination. A delay in attaining maximum levels of allelic Cre-mediated recombination was observed for a mutant defective in telomere clustering. None of the mutants affected ectopic levels of recombination. These data suggest that stable, close homolog juxtaposition in yeast is distinct from pre-DSB pairing interactions, requires both DSB and SEI formation, but does not depend on crossovers or SC.


Regulation of angiotensin II AT1 and AT2 receptors in neonatal ureteral obstruction.
Chronic unilateral ureteral obstruction (UUO) in early development activates the intrarenal renin-angiotensin system and leads to profound renal vasoconstriction, renal growth arrest, and interstitial fibrosis. To investigate the response of the AT1 and AT2 subtypes of the angiotensin II (ANG II) receptors to UUO, Sprague-Dawley rats underwent UUO or control sham operation in the first 48 h of life and were studied 1-28 days later. Renal mRNA for renin, AT1 and AT2 receptor, and receptor binding and distribution were determined. In contrast to controls, renin mRNA increased from 14 to 28 days in the obstructed kidney. After ipsilateral UUO, AT1 mRNA was suppressed at 1 day, but had increased compared with controls at 28 days. AT2 receptor mRNA fell rapidly in all kidneys from 1 to 3 days of age, after which it remained undetectable. Compared with the intact opposite kidney, AT2 mRNA was suppressed in the obstructed kidney 1 day after UUO. Compared with controls, AT1 and AT2 receptor binding was decreased by ipsilateral UUO at 1 day, whereas AT1 binding was increased at 28 days. Renal ANG II content was increased in the obstructed compared with the intact opposite kidney 28 days after UUO. In view of the increase in renal renin and angiotensin II production resulting from UUO, increased renal AT1 mRNA and receptor binding are likely to contribute to the vasoconstriction and interstitial fibrosis of the neonatal kidney after prolonged UUO.


Fear in dreams and in wakefulness: Evidence for day/night affective homeostasis.
Recent neuroscientific theories have proposed that emotions experienced in dreams contribute to the resolution of emotional distress and preparation for future affective reactions. We addressed one emerging prediction, namely that experiencing fear in dreams is associated with more adapted responses to threatening signals during wakefulness. Using a stepwise approach across two studies, we identified brain regions activated when experiencing fear in dreams and showed that frightening dreams modulated the response of these same regions to threatening stimuli during wakefulness. Specifically, in Study 1, we performed serial awakenings in 18 participants recorded throughout the night with high-density electroencephalography (EEG) and asked them whether they experienced any fear in their dreams. Insula and midcingulate cortex activity increased for dreams containing fear. In Study 2, we tested 89 participants and found that those reporting higher incidence of fear in their dreams showed reduced emotional arousal and fMRI response to fear-eliciting stimuli in the insula, amygdala and midcingulate cortex, while awake. Consistent with better emotion regulation processes, the same participants displayed increased medial prefrontal cortex activity. These findings support that emotions in dreams and wakefulness engage similar neural substrates, and substantiate a link between emotional processes occurring during sleep and emotional brain functions during wakefulness.


Association of pain with frequency and magnitude of knee loading in knee osteoarthritis.
Although the relationship between pain and the magnitude of medial knee loading has been previously studied, the contribution of frequency of loading has not. The objective of this study was to determine whether the addition of loading frequency (steps/day) to loading magnitude (knee adduction moment [KAM] impulse) helps explain variance in knee pain in people with knee osteoarthritis (OA). Participants were adults with symptomatic knee OA with radiographic signs in the medial knee compartment (n = 38, 10 women). Pain was measured using the pain subscale of the Knee Injury and Osteoarthritis Outcome Score. Participants wore an accelerometer for 1 week to determine the average number of steps/day. The external KAM impulse was calculated from 3-dimensional gait analysis as participants ambulated at self-selected speeds. Knee extensor strength was measured with an isokinetic dynamometer. Linear regression was used to examine the relationship between pain and steps/day after controlling for the KAM impulse, knee extensor strength, and body mass index (BMI). After controlling for BMI (R(2) = 0.02), knee extensor strength (R2 change = 0.26, P < 0.05), and KAM impulse (R2 change = 0.11, P < 0.05), steps/day contributed an additional 9% of variance in pain (P < 0.05). This model accounted for a total of 49% of the variance in pain (F[4,33] = 7.77, P < 0.05). Increased knee loading frequency and magnitude were associated with increased pain. Objective measures of loading frequency should be considered when investigating the incidence and progression of knee OA.


Mechanism of protein-guided folding of the active site U2/U6 RNA during spliceosome activation.
Spliceosome activation involves extensive protein and RNA rearrangements that lead to formation of a catalytically active U2/U6 RNA structure. At present, little is known about the assembly pathway of the latter and the mechanism whereby proteins aid its proper folding. Here, we report the cryo-electron microscopy structures of two human, activated spliceosome precursors (that is, pre-Bact complexes) at core resolutions of 3.9 and 4.2 angstroms. These structures elucidate the order of the numerous protein exchanges that occur during activation, the mutually exclusive interactions that ensure the correct order of ribonucleoprotein rearrangements needed to form the U2/U6 catalytic RNA, and the stepwise folding pathway of the latter. Structural comparisons with mature Bact complexes reveal the molecular mechanism whereby a conformational change in the scaffold protein PRP8 facilitates final three-dimensional folding of the U2/U6 catalytic RNA.


[The functional colonic sphincter in the rat].
In the proximal colon of the rat typically aligned mucosal folds are found which are displaceable but not erasable (transposable folds). The spatial arrangement of these folds produces a tierlike structure in this section of the intestine by the development of functional ring valves, which allow the flow of food in an oral to anal direction. The basis of these mucosal folds is formed by a vascular apparatus which is peculiar to them. The functional unit of the folds and their vascular apparatus is referred to as the 'angioplicate lock system'. A reversal of operation of even the smallest sections of this lock system inevitably produces a fatal ileus.


Deep sequencing of New World screw-worm transcripts to discover genes involved in insecticide resistance.
The New World screw-worm (NWS), Cochliomyia hominivorax, is one of the most important myiasis-causing flies, causing severe losses to the livestock industry. In its current geographical distribution, this species has been controlled by the application of insecticides, mainly organophosphate (OP) compounds, but a number of lineages have been identified that are resistant to such chemicals. Despite its economic importance, only limited genetic information is available for the NWS. Here, as a part of an effort to characterize the C. hominivorax genome and identify putative genes involved in insecticide resistance, we sampled its transcriptome by deep sequencing of polyadenylated transcripts using the 454 sequencing technology. Deep sequencing on the 454 platform of three normalized libraries (larval, adult male and adult female) generated a total of 548,940 reads. Eighteen candidate genes coding for three metabolic detoxification enzyme families, cytochrome P450 monooxygenases, glutathione S-transferases and carboxyl/cholinesterases were selected and gene expression levels were measured using quantitative real-time polymerase chain reaction (qRT-PCR). Of the investigated candidates, only one gene was expressed differently between control and resistant larvae with, at least, a 10-fold down-regulation in the resistant larvae. The presence of mutations in the acetylcholinesterase (target site) and carboxylesterase E3 genes was investigated and all of the resistant flies presented E3 mutations previously associated with insecticide resistance. Here, we provided the largest database of NWS expressed sequence tags that is an important resource, not only for further studies on the molecular basis of the OP resistance in NWS fly, but also for functional and comparative studies among Calliphoridae flies. Among our candidates, only one gene was found differentially expressed in resistant individuals, and its role on insecticide resistance should be further investigated. Furthermore, the absence of mutations in the OP target site and the high frequency of mutant carboxylesterase E3 indicate that metabolic resistance mechanisms have evolved predominantly in this species.


Treatment of acute otitis media: are children entered into clinical trials representative?
This study involved a meta-analysis of randomised control studies of the treatment of acute otitis media and judged the characteristics of these trials on the basis of methods, age groups, interventions, and outcomes. An investigation of the eight trials reported in the meta-analysis of clinical trials revealed that the number of children entered per doctor in all practices was low and that the recruitment rate was probably lower than 44%. Low recruitment rates indicate that the type of children entered into trials may only relate to those with mild to moderate symptoms and signs, which could explain why the results of these trials show that antibiotics are not essential in treating acute otitis media.


Assigning breed origin to alleles in crossbred animals.
For some species, animal production systems are based on the use of crossbreeding to take advantage of the increased performance of crossbred compared to purebred animals. Effects of single nucleotide polymorphisms (SNPs) may differ between purebred and crossbred animals for several reasons: (1) differences in linkage disequilibrium between SNP alleles and a quantitative trait locus; (2) differences in genetic backgrounds (e.g., dominance and epistatic interactions); and (3) differences in environmental conditions, which result in genotype-by-environment interactions. Thus, SNP effects may be breed-specific, which has led to the development of genomic evaluations for crossbred performance that take such effects into account. However, to estimate breed-specific effects, it is necessary to know breed origin of alleles in crossbred animals. Therefore, our aim was to develop an approach for assigning breed origin to alleles of crossbred animals (termed BOA) without information on pedigree and to study its accuracy by considering various factors, including distance between breeds. The BOA approach consists of: (1) phasing genotypes of purebred and crossbred animals; (2) assigning breed origin to phased haplotypes; and (3) assigning breed origin to alleles of crossbred animals based on a library of assigned haplotypes, the breed composition of crossbred animals, and their SNP genotypes. The accuracy of allele assignments was determined for simulated datasets that include crosses between closely-related, distantly-related and unrelated breeds. Across these scenarios, the percentage of alleles of a crossbred animal that were correctly assigned to their breed origin was greater than 90 %, and increased with increasing distance between breeds, while the percentage of incorrectly assigned alleles was always less than 2 %. For the remaining alleles, i.e. 0 to 10 % of all alleles of a crossbred animal, breed origin could not be assigned. The BOA approach accurately assigns breed origin to alleles of crossbred animals, even if their pedigree is not recorded.


Prospective cohort study of neoadjuvant treatment in conservative surgery of soft tissue sarcomas.
1994 marked a decade since the inception of a prospective population-based study on the value of neoadjuvant approach for soft tissue sarcomas of head, neck, and limbs at the Tom Baker Cancer Centre, Calgary, Alberta. To date, 42 patients have been followed for a minimum of 5 years or until death. Each patient received a protocol of 60 mg to 90 mg of Adriamycin infused intra-arterially or intravenously over 3 days into a vessel feeding the involved area, 30 Gy of radiotherapy given over 10 days, and complete resection of the sarcoma 4 to 6 weeks later. The lower dose was used empirically for smaller limbs (e.g., arm). Two of the 42 patients were immediate failures of protocol, with one requiring amputation and one requiring later reexcision. In the 38 appendicular lesions, the ultimate limb salvage rate was 97.5%. All tumors were associated with a high risk of local recurrence with 15 being previous local failures. The rest were deep and grade 2 or 3 lesions. Serious local complications were seen in one patient (2.5%) who had wound necrosis requiring reoperation. Minor wound complications were seen in five patients (12.5%) (one wound infection, one resolved edema, three long-term drainage). There was one local recurrence; thus 5-year local control was 97%. No patient had long-term morbidity related to the treatment. No effect on systemic control was suggested. Our report demonstrates that this combined modality approach provides superior local control of soft tissue sarcomas with low postoperative morbidity.


Alterations in predicted growth rates of pediatric kidneys treated with extracorporeal shockwave lithotripsy.
The long-term effects of extracorporeal shockwave lithotripsy (SWL) on the kidneys of children treated for renal calculi are unclear. In order to determine if SWL has any negative effects on renal growth rates, we reviewed long-term (mean 9-year) follow-up data on 29 pediatric patients treated between 1984 and 1988 with an unmodified Dornier HM3 lithotripter. Changes in renal length, serum creatinine, and blood pressure were analyzed. Predicted renal growth was calculated using a formula for age-adjusted renal length. Treated kidneys were stratified into normal and abnormal groups based on a history of renal surgery, evidence of recurrent infection, and obvious anatomic abnormalities. Fifty-six upper urinary tract calculi were treated in 34 renal units. Twenty-two renal units (68%) were rendered stone free, and 65% of the patients continue to be stone free. At follow-up, one patient was classified as having new-onset hypertension, and the mean serum creatinine was 0.93 +/- 0.08 mg/dL. Both at treatment and at follow-up, no significant differences were found in the sizes of the treated and untreated kidneys. However, at treatment, the abnormal group of kidneys seemed to be smaller than expected (mean Z -1.30 +/- 1.10), whereas the group of normal kidneys was very close (mean Z 0.18 +/- 0.54) to the predicted length. At follow-up, the deviations between actual and predicted renal length were significantly more negative. Treated kidneys were an additional 1.26 +/- 0.49 SD units below their expected length (p = 0.02). Untreated kidneys were further below normal as well but possibly to a lesser degree (-0.82 +/- 0.36; p <0.04). Although there was a trend for the abnormal group to have smaller kidneys than the normal group, both groups showed the same trend toward an age-adjusted reduction in renal growth at follow-up. The alterations in renal growth patterns observed in this population are unsettling and could be secondary to either treatment effect (SWL) or, more likely, to some underlying pathology intrinsic to pediatric kidneys with urolithiasis. Until further data are available, SWL in the pediatric population should be applied with caution and at the lowest dosage sufficient to achieve stone comminution.


Fossil and Living Conchostracan Distribution in Kansas-Oklahoma across a 2OO-Millon-Year Time Gap.
Fifty-nine of 493 ponds sampled in the Wellington fossil conchostracan belt contained Cyzicus mexicanus (Claus). Persistent habitat preference and faunal association were also found for four orders of insects (Odonata, Ephemeroptera, Neuroptera, and Homoptera). Comparative limnology is detailed. Greater geographic fractionation of Permian conchostracan gene-pools is attributed to a more arid climate indicated by evaporites.


The ProtekDuo as double lumen return cannula in V-VP ECMO configuration: A first-in-man method description.
We present a case of acute respiratory distress syndrome (ARDS) secondary to COVID-19 who required venovenous extracorporeal membrane oxygenation (V-V ECMO). Initially, a right ventricular assist device (RVAD), the ProtekDuo with an oxygenator, was placed in an outside heart center and the patient was transferred to us for ECMO management. Due to severe hypoxia, the configuration was later modified, and a 25 Fr femoral drainage cannula was inserted for venous drainage only. The arterial return tubing was spliced and using a Y-connector, arterialized blood was returned through both limbs of the ProtekDuo resulting in a significantly increased oxygenation and flow.


Biochemical and morpho-mechanical properties, and structural organization of rat tail tendon collagen in diet-induced obesity model.
We have investigated the impact of obesity on the structural organization, morpho-mechanical properties of collagen fibers from rat tail tendon fascicles (RTTFs). Polarized Raman microspectroscopy showed that the collagen bands 855, 875, 938, and 960 cm-1 as well as those 1631 and 1660 cm-1 were affected by diet. Mechanical properties exhibited an increase in the yield strength from control (CTRL) to high fat (HF) diet (9.60 ± 1.71 and 13.09 ± 1.81 MPa) (p < 0.01) and ultimate tensile strength (13.12 ± 2.37 and 18.32 ± 2.83 MPa) (p < 0.05) with no significant change in the Young's Modulus. During mechanical, the band at 875 cm-1 exhibited the most relevant frequency shift (2 cm-1). The intensity of those at 855, 875, and 938 cm-1 in HF collagen displayed a comparable response to mechanical stress as compared to CTRL collagen with no significant diet-related changes in the Full Width at Half Maximum. Second harmonic generation technique revealed i) similar fiber straightness (0.963 ± 0.004 and 0.965 ± 0.003) and ii) significant changes in fibers diameter (1.48 ± 0.07 and 1.52 ± 0.08 μm) (p < 0.05) and length (22.06 ± 2.38 and 29.00 ± 3.76 μm) (p < 0.001) between CTRL and HF diet, respectively. The quantification of advanced glycation end products (AGEs) revealed an increase in both carboxymethyl-lysine and total fluorescence AGEs from CTRL to HF RTTFs.


Retroperitoneal approach for portasystemic decompression.
A left retroperitoneal approach has been developed and used to perform a renal-splenic shunt for portal decompression in seven patients with massive variceal hemorrhage. In each patient, at least one intra-abdominal finding contraindicated a conventional transperitoneal portasystemic shunt. Retroperitoneal approach avoided possible complications and permitted successful portal decompression with cessation of bleeding in each instance. These results support the use of this technique when it is necessary to avoid the peritoneal cavity in a patient requiring portasystemic decompression.


Two Acute Liver Injuries in a Patient With Malnutrition.
Chronic starvation and refeeding have been associated with liver injury (LI). We present a patient with anorexia nervosa who exhibited both phenomena of malnutrition-related LI. At presentation, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were elevated at 154 and 136 U/L, respectively, and rose rapidly to peaks of 750 and 638 U/L, respectively, as nutrition was introduced. Mechanisms of starvation-related LI include impaired degradation and secretion of lipids, as well as starvation-induced autophagy. LI during refeeding may be related to rapid increase in glucose availability. These phenomena are crucial to consider in patients with chronic starvation undergoing refeeding.


Aluminum inhibits the fast phase of voltage-dependent calcium influx into synaptosomes.
Aluminum has been shown to have neurotoxic effects, but the mechanisms by which it acts are not well understood. Because it has been reported that aluminum can interact with Ca2+-binding sites, the possibility that aluminum might interfere with Ca2+ influx into synaptosomes was examined. At concentrations of 50 microM and greater, aluminum significantly inhibited the fast phase (0-1 s) of the voltage-dependent uptake of 45Ca2+ into synaptosomes. Higher concentrations of aluminum also reduced 45Ca2+ uptake measured at 1 s in nondepolarizing media and inhibited the slow phase of 45Ca2+ uptake into synaptosomes whether they were suspended in either low K or high K media. The possibility that aluminum competitively inhibits the fast phase of Ca2+ influx was investigated. Aluminum (250 microM) increased the apparent KT (concentration of Ca2+ at which Ca2+ transport is half maximal) for 45Ca2+ of fast phase voltage-dependent channels and slightly decreased the maximal influx (Jmax). These effects are characteristic of a mixed type inhibitor, and the apparent Ki for Al3+ is estimated to be 0.64 mM. The interaction of aluminum with the fast phase of voltage-dependent calcium influx may disrupt intraneuronal calcium homeostasis and may also represent a means by which aluminum could accumulate intraneuronally.


UPLC-PDA-ESI-QTOF-MS/MS and GC-MS analysis of Iranian Dracocephalum moldavica L.
Dracocephalum moldavica L. is a significant component in the Iranian food basket. This study aimed to investigate the bioactive compounds and biological activities of different extracts obtained from D. moldavica aerial parts. From the aerial parts, a crude methanolic (MeOH) extract and its four sub-fractions, that is, petroleum ether (Pet), ethyl acetate (EtOAc), n-butanol (n-BuOH), and aqueous (water) extracts were obtained. The total phenolic and flavonoid contents as well as the antioxidant and cytotoxic activities of the extracts were determined. Moreover, the phytochemical profiles of the essential oil (EO) and of those extracts with the highest antioxidant activity measured by GC/MS and UPLC-PDA-ESI-QTOF-MS/MS. Results showed that the highest concentrations of phenols and flavonoids as well as the most potent antioxidant potential according to the DPPH method were determined in the EtOAc and MeOH extracts with IC50 values of 22.0 and 34.4 µg.ml-1, respectively. Quantitative analysis of these extracts was subsequently performed by UPLC-PDA-ESI-QTOF-MS/MS. Both extracts contained mainly rosmarinic acid, caffeic acid, and 2-hydroxycinnamic acid, which may be responsible for their high antioxidant activity. Moreover, none of the extracts showed cytotoxic effects against MCF7, SW48, and a normal cell line of mouse embryonic fibroblast cells (NIH/3T3) in the tested concentrations (up to 400 μg.ml-1). Additionally, GC-MS analysis showed that oxygenated monoterpenes (55.4%) were the main constituents of the EO of D. moldavica.


Dynamics of Bacterial Communities Mediating the Treatment of an As-Rich Acid Mine Drainage in a Field Pilot.
Passive treatment based on iron biological oxidation is a promising strategy for Arsenic (As)-rich acid mine drainage (AMD) remediation. In the present study, we characterized by 16S rRNA metabarcoding the bacterial diversity in a field-pilot bioreactor treating extremely As-rich AMD in situ, over a 6 months monitoring period. Inside the bioreactor, the bacterial communities responsible for iron and arsenic removal formed a biofilm ("biogenic precipitate") whose composition varied in time and space. These communities evolved from a structure at first similar to the one of the feed water used as an inoculum to a structure quite similar to the natural biofilm developing in situ in the AMD. Over the monitoring period, iron-oxidizing bacteria always largely dominated the biogenic precipitate, with distinct populations (Gallionella, Ferrovum, Leptospirillum, Acidithiobacillus, Ferritrophicum), whose relative proportions extensively varied among time and space. A spatial structuring was observed inside the trays (arranged in series) composing the bioreactor. This spatial dynamic could be linked to the variation of the physico-chemistry of the AMD water between the raw water entering and the treated water exiting the pilot. According to redundancy analysis (RDA), the following parameters exerted a control on the bacterial communities potentially involved in the water treatment process: dissolved oxygen, temperature, pH, dissolved sulfates, arsenic and Fe(II) concentrations and redox potential. Appreciable arsenite oxidation occurring in the bioreactor could be linked to the stable presence of two distinct monophylogenetic groups of Thiomonas related bacteria. The ubiquity and the physiological diversity of the bacteria identified, as well as the presence of bacteria of biotechnological relevance, suggested that this treatment system could be applied to the treatment of other AMD.


Gene-mediated restoration of cartilage matrix by combination insulin-like growth factor-I/interleukin-1 receptor antagonist therapy.
Combination of growth factor gene-enhanced cartilage matrix synthesis with interleukin-1 receptor antagonist protein (IL-1Ra) abrogation of cartilage matrix degradation may reduce and possibly reverse cartilage loss in synovitis and osteoarthritis. The feasibility of cotransduction of synovial membrane with two such genes that may act on cartilage homeostasis was investigated in an in vitro coculture system. Cultured synoviocytes in monolayer were cotransduced with E1-deleted adenoviral vectors, one containing IGF-I coding sequence under cytomegalovirus (CMV) promoter control (200 multiplicities of infection (moi)), and the second containing IL-1Ra sequence under CMV promoter control (100 moi). Adenovirus-IGF-I (AdIGF-I) transduction and AdIGF-I/AdIL-1Ra cotransduction of synovial monolayer cultures resulted in increased IGF-I mRNA and ligand expression, and similarly AdIL-1Ra and AdIGF-I/AdIL-1Ra-transduced cultures expressed high levels of IL-1Ra. Northern analysis confirmed a single mRNA transcript of the appropriate size for both IGF-I and IL-1Ra transgene expression. Synovial cell monolayer and cartilage explant coculture experiments were used to examine the effects of IGF-I and IL-1Ra protein expressed by transduced synoviocytes on normal and IL-1-depleted cartilage. Transduced monolayer cultures produced peak medium IGF-I content of 114+/-20.2 ng/ml and IL-1Ra levels of 241.8+/-10.5 ng/ml at 48 h after transduction. These IGF-I concentrations were sufficient to produce significantly increased proteoglycan (PG) content of normal cartilage cultured in medium conditioned by AdIGF-I and AdIGF-I/AdIL-1Ra-transduced synoviocytes. Interleukin-1-exposed cartilage was markedly depleted of PG, and this catabolic state was partially reversed in AdIGF-I-transduced cultures and fully reversed by AdIGF-I/AdIL-1Ra-transduced synovial cocultures. These data indicate that cultured synoviocytes are readily cotransduced by two recombinant adenoviral vectors containing transgenes active in restoring joint health. The AdIL-1Ra and AdIGF-I transgenes were abundantly expressed and the secreted products achieved therapeutic concentrations by day 2. The resulting increase in matrix biosynthesis returned cartilage PG content to normal levels. These data suggest that there may be significant value in cotransduction of synovial membrane to attenuate cartilage malacia associated with synovitis, injury, or early arthritis.


Combination Treatment of Withalongolide a Triacetate with Cisplatin Induces Apoptosis by Targeting Translational Initiation, Migration, and Epithelial to Mesenchymal Transition in Head and Neck Squamous Cell Carcinoma.
Treatment regimens for head and neck squamous cell carcinoma (HNSCC) typically include cisplatin and radiotherapy and are limited by toxicities. We have identified naturally derived withalongolide A triacetate (WGA-TA) from Physalis longifolia as a lead compound for targeting HNSCC. We hypothesized that combining WGA-TA with cisplatin may allow for lower, less toxic cisplatin doses. HNSCC cell lines were treated with WGA-TA and cisplatin. After treatment with the drugs, the cell viability was determined by MTS assay. The combination index was calculated using CompuSyn. The expression of proteins involved in the targeting of translational initiation complex, epithelial to mesenchymal transition (EMT), and apoptosis were measured by western blot. Invasion and migration were measured using the Boyden-chamber assay. Treatment of MDA-1986 and UMSCC-22B cell lines with either WGA-TA or cisplatin alone for 72 h resulted in a dose dependent decrease in cell viability. Cisplatin in combination with WGA-TA resulted in significant synergistic cell death starting from 1.25 μM cisplatin. Combination treatment with WGA-TA resulted in lower cisplatin dosing while maintaining the downregulation of translational initiation complex proteins, the induction of apoptosis, and the blockade of migration, invasion, and EMT transition. These results suggest that combining a low concentration of cisplatin with WGA-TA may provide a safer, more effective therapeutic option for HNSCC that warrants translational validation.


Mobilization of peripheral-blood stem cells by concurrent administration of daniplestim and granulocyte colony-stimulating factor in patients with breast cancer or lymphoma.
To evaluate the safety and hematopoietic activity of daniplestim administered concurrently with granulocyte colony-stimulating factor (G-CSF) for peripheral-blood stem-cell (PBSC) mobilization. In the initial dose-escalation phase, 25 patients with adenocarcinoma of the breast (AB; 13 patients) or lymphoma (12 patients) were given daniplestim at doses ranging from 0.1 to 3.75 microgram/kg/d plus G-CSF 10 microgram/kg/d. In the randomized phase, 52 patients with AB (27 patients) or lymphoma (25 patients) were randomized within disease categories to the daniplestim dose chosen in the dose-escalation phase plus G-CSF 10 microgram/kg/d (D+G) or placebo plus G-CSF 10 microgram/kg/d (P+G) for up to 7 days. A daniplestim dose of 2. 5 microg/kg/d was chosen for further study because it was hematopoietically active and had an acceptable side-effect profile. In the randomized phase, in patients with AB, D+G was associated with a higher probability (P =.0696) of collecting >/= 2.5 x 10(6) CD34(+) cells/kg and significantly higher circulating CD34(+) cell counts (P =.0498) on days 6 through 9 after the initiation of dosing. The target level was more likely to be reached with additional leukaphereses in the patients given D+G. Patients given P+G did not benefit from additional leukaphereses beyond the first procedure. The type of mobilization did show a trend toward a shorter duration of neutropenia in the D+G group. The adverse events with D+G consisted largely of mild to moderate flu-like symptoms, including headache and fever, and occurred more frequently than with P+G. Daniplestim administered at 2.5 microgram/kg/d is tolerable and active when combined with G-CSF, and the combination may prove more effective than G-CSF alone in promoting the collection of adequate numbers of CD34(+) cells for PBSC infusion in patients with AB.


Biologic banking in cohort studies, with special reference to blood.
Those who conduct cohort studies in cancer epidemiology increasingly use biochemical analyses as an important component. Some of the potentially important considerations when banked blood is used include the conditions and temperature of storage, effects of thawing, and the stability of specific substances under prolonged subfreezing temperatures. I have reviewed a selected number of biochemical substances.


Comparison of hamstring/quadriceps isokinetic strength ratios and power in tennis, squash and track athletes.
Isokinetic assessment of elite squash, tennis and track athletes confirms the accepted ratio of 60 to 80 per cent hamstring to quadriceps when testing at 90 deg-sec-1 for peak strength (torque). However, significant variations occur at higher test speeds up to 300 deg.sec-1 with the hamstrings becoming more prominent especially (p less than 0.001) in the nonpreferred (NP) leg. There was no significant difference between sports, and wide individual differences occurred. Analysis of (work.sec-1.kg-1 body weight) power showed a significantly higher work output (p less than 0.01) by track athletes than squash and tennis players, but, unlike hamstring/quadriceps ratio, no significant difference between preferred (P) and nonpreferred leg. The maximum power output was achieved around 220 to 250 deg.sec-1. Power between preferred and nonpreferred legs was the same but the torque ratio differed indicating that the hamstrings provided proportionately more work in the NP leg at higher speeds.


Oral bisphosphonate therapy for vitamin D intoxication of the infant.
Vitamin D intoxication in infancy has serious consequences attributable to acute hypercalcemia and subsequent hypercalcuria/nephrocalcinosis. Current treatments of patients with vitamin D intoxication are unsatisfactory and associated with prolonged hypercalcemia. We now report the use of oral alendronate sodium in a 3-month-old infant with vitamin D intoxication. Short-term oral alendronate sodium treatment effectively corrected hypercalcemia/hypercalciuria, decreased the duration of hospitalization, and appears safe in 15 months of observation.


The impact of COVID-19 on surgical training: a systematic review.
Coronavirus disease (COVID-19) has caused global disruption to health care. Non-urgent elective surgical cases have been cancelled, outpatient clinics have reduced and there has been a reduction in the number of patients presenting as an emergency. These factors will drastically affect the training opportunities of surgical trainees. The aim of this systematic review is to describe the impact of COVID-19 on surgical training globally. The review was performed in line with Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines and registered with the Open Science Framework (OSF). Medline, EMBASE, PubMed and the Cochrane Central Register of Controlled Trials were searched. The searches identified 499 articles, 29 of which were included in the review. This contained data from more than 20 countries with 5260 trainees and 339 programme directors. Redeployment to non-surgical roles varied across studies from 6% to 35.1%. According to all of the studies, operative experience has been reduced. Knowledge learning had been switched to online platforms across 17 of the studies and 7 reported trainees had increased time to devote to educational/academic activities. All of the studies reporting on mental health report negative associations with increased stress, ranging from 54.9% to 91.6% of trainees. The impact of COVID-19 on surgical trainees has been experienced globally and across all specialities. Negative effects are not limited to operative and clinical experience, but also the mental health and wellbeing of trainees. Delivery of surgical training will need to move away from traditional models of learning to ensure trainees are competent and well supported.


The activity of the stapedius muscle in man during vocalization.
The EMG of the stapedius muscle and visible movements in the stapedius tendon during the subject's own vocalization of an [a:] and during contralateral acoustic stimulation were studied in subjects with eardrum perforation. The threshold for stapedius activity was near the lowest vocal intensity that the subjects could produce. At normal vocal effort the stapedius muscle was activated to about 50% of its maximum value. The EMG often started before the vocal sound, indicating that the stapedius muscle can be activated from the central nervous system as a part of the vocalization process. It is suggested that the contraction of the stapedius muscle during vocalization reduces the masking caused by the low frequency components of the person's own voice even at normal vocal effort and thereby improves intelligibility of simultaneous external speech.


Inhibition of miR-495 Improves Both Vascular Remodeling and Angiogenesis in Pulmonary Hypertension.
Pulmonary hypertension (PH) is a chronic progressing vascular disease characterized by pulmonary arteriole remodeling and loss of pulmonary microvasculature. The aim of this study was to investigate a potential role for the miR-495 in PH pathogenesis and to explore its therapeutic potential in PH. Male C57BL/6J mice were injected with SU5416 weekly during 3 weeks of exposure to 10% oxygen to cause PH. We first tested the effects of adeno-associated virus 9 (AAV9) delivery which was specifically designed to block miR-495 in the lungs of the PH model. Then, the biological function of miR-495 was analyzed in cultured pulmonary arterial endothelial cells (PAECs) under hypoxic condition. The inhibition of miR-495 improves hemodynamics and vascular remodeling in PH. At the same time, these effects were associated with increases in angiogenic transcription factor VEZF1 and marked upregulation of other angiogenic genes such as Angpt-1 and IGF1. In vitro, cultured mouse PAECs were transfected with miR-495 inhibitor or miR-495 mimics. Both the flow cytometry results and CCK8 assay showed that miR-495 inhibitor increased the percentage of cells in the G2/M+S phase, and the wound healing assays indicated that the migration capacity of PAECs transfected with miR-495 inhibitor was increased compared to the inhibitor-NC cells. Our results indicate that AAV9-TuD-miR-495 delivery improves hemodynamic and pulmonary vascular structural changes in PH mice.


Differential optical microscopy based on higher-order Gaussian-Hermite beam patterns.
A new method for performing scanning differential optical microscopy is described. This method is based on the use of a higher-order transverse mode of a laser. The technique is capable of responding to both amplitude and phase structures on the object. The sensitivity of the system in detecting differential object phase structures is similar to that of a dc scanning Nomarski microscope. The system is employed to image the surface structure of an integrated circuit.