Two distinct mechanisms for redistribution of lymphocyte surface macromolecules. II. Contrasting effects of local anesthetics and a calcium ionophore.
In the previous study, lymphocyte surface molecules were separated into two subsets depending on whether capping was associated was associated with redistribution of cytoplasmic myosin. In the present study, the effects of the local anesthetic chlorpromazine and of the Ca2+ ionophore A23187 were compared. Both drugs affected the surface redistribution of immunoglobulin (Ig), Fc receptors, and the TL antigen--molecules that appear to cap by association with microfilaments--but had no effect on the Thy.1 (theta) and H2 antigens--molecules that cap slowly, apparently unlinked to microfilament function. The capping of Ig, Fc receptor, and TL was inhibited while that of H2 and theta was not. Both drugs reversed the Ig Fc receptor, and TL caps but not the H2 and theta caps. In the former group, the reversal of caps was accompanied by a parallel reversal of the myosin segregated to the cap area. The appearance of myosin after drug treatment varied: chlorpromazine resulted in a diffuse pattern similar to that of normal lymphocytes, whereas A23187 produced an array of aggregates and coarse filaments. The results are compatible with the view that two mechanisms for capping exist in the lymphocyte. The Ca2+ ionophore may affect capping of microfilament-dependent caps by producing a systemic activation of contractile proteins while chlorpromazine may act by disrupting a Ca2+-dependent link between surface complexes and the contractile proteins.


Therapeutic effects of serelaxin in acute heart failure.
Over the past few decades, research on the peptide hormone, relaxin, has significantly improved our understanding of its biological actions under physiological and diseased conditions. This has facilitated the conducting of clinical trials to explore the use of serelaxin (human recombinant relaxin). Acute heart failure (AHF) is a very difficult to treat clinical entity, with limited success so far in developing new drugs to combat it. A recent phase-III RELAX-AHF trial using serelaxin therapy given during hospitalization revealed acute (ameliorated dyspnea) and chronic (improved 180-day survival) effects. Although these findings support a substantial improvement by serelaxin therapy over currently available therapies for AHF, they also raise key questions and stimulate new hypotheses. To facilitate the development of serelaxin as a new drug for heart disease, joint efforts of clinicians, research scientists and pharmacological industries are necessary to study these questions and hypotheses. In this review, after providing a brief summary of clinical findings and the pathophysiology of AHF, we present a working hypothesis of the mechanisms responsible for the observed efficacy of serelaxin in AHF patients. The existing clinical and preclinical data supporting our hypotheses are summarized and discussed. The development of serelaxin as a drug provides an excellent example of the bilateral nature of translational research.


Foveal cone function in nonproliferative diabetic retinopathy and macular edema.
To study results of foveal cone electroretinography (ERG) in patients with nonproliferative diabetic retinopathy (NPDR), with and without clinically significant macular edema (CSME). Electroretinograms of foveal cones were elicited, with a dual-beam stimulator-ophthalmoscope, from 18 consecutive patients with bilateral NPDR and unilateral CSME. Analysis of resulting data revealed that mean amplitude was significantly lower in eyes with CSME and in eyes without CSME., compared with that in normal eyes. Mean implicit time was significantly longer in eyes with CSME compared with that in normal eyes, but in eyes of diabetic patients without CSME., mean implicit time was the same as that in normal eyes. Amplitudes were directly correlated and implicit times were inversely correlated with best-corrected Snellen visual acuity in eyes with and without CSME. Eyes with CSME had significantly lower amplitudes and longer implicit times than their fellow eyes without CSME. In addition, eyes with NPDR, with or without CSME, did not exhibit the normal rise in amplitude with increasing duration of light exposure. Electroretinographic findings showed abnormalities in foveal cone responses in eyes with NPDR, particularly in the presence of CSME. These results may support a functional role for outer retinal dysfunction in patients with diabetic retinopathy and loss of central vision.


Methyl-CpG Binding Protein 2 (Mecp2) Regulates Sensory Function Through Sema5b and Robo2.
Mutations in the gene encoding the MECP2 underlies Rett syndrome, a neurodevelopmental disorder in young females. Although reduced pain sensitivity in Rett syndrome patients and in partial MeCP2 deficient mice had been reported, these previous studies focused predominantly on motor impairments. Therefore, it is still unknown how MeCP2 is involved in these sensory defects. In addition, the human disease manifestations where males with mutations in MECP2 gene normally do not survive and females show typical neurological symptoms only after 18 months of age, is profoundly different in MeCP2-deficient mouse where all animals survived, and males but not females displayed Rett syndrome phenotypes at an early age. Thus, the mecp2-deficient zebrafish serves as an additional animal model to aid in deciphering the role and mechanisms of Mecp2 in neurodevelopment. Here, we used two independent methods of silencing expression of Mecp2 in zebrafish to uncover a novel role of Mecp2 in trigeminal ganglion sensory neurons during the embryonic development. mecp2-null mutation and morpholino-mediated silencing of Mecp2 in the zebrafish embryos resulted in defects in peripheral innervation of trigeminal sensory neurons and consequently affecting the sensory function. These defects were demonstrated to be dependent on the expression of Sema5b and Robo2. The expression of both proteins together could better overcome the defects caused by Mecp2 deficiency as compared to the expression of either Sema5b or Robo2 alone. Sema5b and Robo2 were downregulated upon Mecp2 silencing or in mecp2-null embryos, and Chromatin immunoprecipitation (ChIP) assay using antibody against Mecp2 was able to pull down specific regions of both Sema5b and Robo2 promoters, showing interaction between Mecp2 and the promoters of both genes. In addition, cell-specific expression of Mecp2 can overcome the innervation and sensory response defects in Mecp2 morphants indicating that these MeCP2-mediated defects are cell-autonomous. The sensory deficits caused by Mecp2 deficiency mirror the diminished sensory response observed in Rett syndrome patients. This suggests that zebrafish could be an unconventional but useful model for this disorder manifesting defects that are not easily studied in full using rodent models.


Mass Spectrometry in Proteomics: Technologies, Methods, and Research Applications for the Life Sciences.
Mass spectrometry is a powerful tool in the hand of life science researchers, who constantly develop and apply new methods for the investigation of biomolecules, such as proteins, peptides, metabolites, lipids, and glycans. In this review, we will discuss the importance of mass spectrometry for the life science sector, with a special focus on the most relevant current applications in the field of proteomics. Moreover, we will comment on the factors that research groups should consider when setting up a mass spectrometry laboratory, and on the fundamental role played by academic core facilities and industrial service providers.


Slightly elevated high-sensitivity C-reactive protein (hsCRP) concentrations are associated with carotid atherosclerosis in women with varying degrees of glucose tolerance.
We examined whether high-sensitivity C-reactive protein (hsCRP) ≥2.0 mg/L was associated with increased intima-media thickness (IMT), plaque burden, and plaque echolucency in carotid arteries. Women (n = 635) from a population sample of 64-year-old females with varying degrees of glucose tolerance underwent risk factor assessment, measurement of hsCRP, and ultrasound examinations of the carotid arteries. Participants with hsCRP levels ≥2.0 mg/L had elevated carotid bulb IMT independently of other cardiovascular risk factors compared with those with hsCRP <2.0 mg/L. The participants with plaques in the highhsCRP group had larger total plaque area compared to those with plaque in the lower hsCRP group. Plaque echolucency did not differ between groups. High-sensitivity CRP levels ≥2.0 mg/L were accompanied by elevated IMT in the carotid bulbs independently of other cardiovascular risk factors. Total plaque area was larger among women with plaques in the high hsCRP group versus the lower hsCRP group.


Increased incidence of VTE in sickle cell disease patients: risk factors, recurrence and impact on mortality.
Previous reports show increased incidence of venous thromboembolism [VTE, deep-vein thrombosis (DVT) and pulmonary embolus (PE)] in sickle cell disease (SCD) patients but did not account for frequency of hospitalization. We determined the incidence of VTE in a SCD cohort versus matched controls. For SCD patients, risk factors for incident VTE, recurrence and the impact on mortality were also determined. Among 6237 patients with SCD, 696 patients (11·2%) developed incident-VTE: 358 (51·6%) had PE (±DVT); 179 (25·7%) had lower-extremity DVT only and 158 (22·7%) had upper-extremity DVT. By 40 years of age, the cumulative incidence of VTE was 17·1% for severe SCD patients (hospitalized ≥3 times a year) versus 8·0% for the matched asthma controls. Amongst SCD patients, women (Hazard ratio [HR] = 1·22; 95% confidence interval [CI]: 1·05-1·43) and those with severe disease (HR = 2·86; 95% CI: 2·42-3·37) had an increased risk of VTE. Five-year recurrence was 36·8% in patients with severe SCD. VTE was associated with increased risk of death (HR = 2·88, 95% CI: 2·35-3·52). In this population-based study, the incidence of VTE was higher in SCD patients than matched controls and was associated with increased mortality. The high incidence of recurrent VTE in patients with severe SCD suggests that extended anticoagulation may be indicated.


Mechanisms of Ca2+ antagonism in imipramine-induced toxicity of isolated adult rat cardiomyocytes.
Classic explanations of cyclic antidepressant toxicity often focus on Na+ channel blockade; however, cyclic antidepressant toxicity often causes decreased myocardial contractile function. The present experiments first examine inhibition of cytosolic Ca2+ signals by imipramine. Second, the experiments test if alkalinization prevents the inhibition of Ca2+ signals. Cardiomyocytes from adult rat hearts were loaded with fura-2 dye, and intracellular calcium, [Ca2+]i, was quantified using ratio fluorescence techniques. Changes in [Ca2+]I were induced by electrical pacing, depolarization with KCl (84 mM), or treatment with caffeine (10 mM). Imipramine (10-30 microM) inhibited [Ca2+]i transients in electrically paced cardiomyocytes. Imipramine (7.5-30 microM) also inhibited Ca2+ signals in KCl depolarized cells. These inhibitory effects were similar to those observed with nisoldipine (100-2000 nM), a selective L-channel blocker. The rise in [Ca2+]i that was triggered with caffeine (10 mM) was not significantly changed by imipramine (30 microM). Inhibition of KCl-induced Ca2+ signals by imipramine was prevented by alkalinization of the medium (tris(hydroxymethyl)aminomethane, pH 7.6), but not by elevation of extracellular sodium to 170 mM. Alkalinization was effective in the presence of HOE642, a selective Na+/H+ (NHE) subtype 1 inhibitor. These data show that imipramine causes Ca2+ antagonism in heart cells which is independent of sarcoplasmic reticulum Ca2+, and that alkaline treatment prevents this Ca2+ antagonism rather than stimulating an alternate source of Ca2+ via Na+/H+ and subsequent Na+/Ca2+ exchange.


The human phosphotyrosine signaling network: evolution and hotspots of hijacking in cancer.
Phosphotyrosine (pTyr) signaling, which plays a central role in cell-cell and cell-environment interactions, has been considered to be an evolutionary innovation in multicellular metazoans. However, neither the emergence nor the evolution of the human pTyr signaling system is currently understood. Tyrosine kinase (TK) circuits, each of which consists of a TK writer, a kinase substrate, and a related reader, such as Src homology (SH) 2 domains and pTyr-binding (PTB) domains, comprise the core machinery of the pTyr signaling network. In this study, we analyzed the evolutionary trajectories of 583 literature-derived and 50,000 computationally predicted human TK circuits in 19 representative eukaryotic species and assigned their evolutionary origins. We found that human TK circuits for intracellular pTyr signaling originated largely from primitive organisms, whereas the inter- or extracellular signaling circuits experienced significant expansion in the bilaterian lineage through the "back-wiring" of newly evolved kinases to primitive substrates and SH2/PTB domains. Conversely, the TK circuits that are involved in tissue-specific signaling evolved mainly in vertebrates by the back-wiring of vertebrate substrates to primitive kinases and SH2/PTB domains. Importantly, we found that cancer signaling preferentially employs the pTyr sites, which are linked to more TK circuits. Our work provides insights into the evolutionary paths of the human pTyr signaling circuits and suggests the use of a network approach for cancer intervention through the targeting of key pTyr sites and their associated signaling hubs in the network.


Estimates of diffuse phosphorus sources in surface waters of the United States using a spatially referenced watershed model.
The statistical watershed model SPARROW (SPAtially Referenced Regression On Watershed attributes) was used to estimate the sources and transport of total phosphorus (TP) in surface waters of the United States. We calibrated the model using stream measurements of TP from 336 watersheds of mixed land use and spatial data on topography, soils, stream hydrography, and land use (agriculture, forest, shrub/grass, urban). The model explained 87% of the spatial variability in log transformed stream TP flux (kg yr(-1)). Predictions of stream yield (kg ha(-1) yr(-1)) were typically within 45% of the observed values at the monitoring sites. The model identified appreciable effects of soils, streams, and reservoirs on TP transport. The estimated aquatic rates of phosphorus removal declined with increasing stream size and rates of water flushing in reservoirs (i.e. areal hydraulic loads). A phosphorus budget for the 2.9 million km2 Mississippi River Basin provides a detailed accounting of TP delivery to streams, the removal of TP in surface waters, and the stream export of TP from major interior watersheds for sources associated with each land-use type.


Comparative Results Between "Epi-Off" Accelerated and "Epi-Off" Standard Corneal Collagen Crosslinking-UVA in Progressive Keratoconus - 7 Years of Follow-Up.
The purpose of the present study was to assess the long-term efficiency and safety of the "epi-off" accelerated CXL (9 mW/cm2 for 10 minutes) in comparison to the standard "epi-off" CXL (3 mW/cm2 for 30 minutes) in terms of topographical and keratometric parameters, refractive data and visual outcomes at 7 years of follow-up, in progressive keratoconus. A retrospective and comparative study was performed. A total of 183 eyes from 183 patients with documented progressive keratoconus were included in the study. The patients were divided in two groups: 93 eyes from 93 patients underwent "epi-off" standard cross-linking technique (3 mW/cm2 for 30 minutes) (S-CXL group) and 90 eyes from 90 patients underwent accelerated "epi-off" corneal CXL technique (9 mW/cm2 for 10 minutes) (A-CXL group). Improvements in uncorrected distance visual acuity (UDVA) were statistically significant compared to baseline values in both groups at each time-point visit (p=0.0421 at 1 year, p=0.0411 at 7 years for A-CXL and p=0.0375 at 1 year, p=0.0389 at 7 years for S-CXL). At 7 years there was a statistically significant increase in CDVA (p=0.039 in the A-CXL group and p=0.0343 in the S-CXL group at 7 years). Statistically significant reduction was noticed in Ksteep (p=0.0411 in A-CXL group and p=0.0224 in S-CXL group), Kflat (p=0.0198 in A-CXL group and p=0.008 in S-CXL group), K mean (p=0.0106 in A-CXL group and p=0.0193 in S-CXL group) and Kmax (p=0.0413 in A-CXL group and p=0.054 in S-CXL group) at 7 years, compared to baseline values, in both groups, but without any statistically difference between the two procedures, at all time-point visits (p>0.05). The long-term outcomes of "epi-off" accelerated corneal collagen crosslinking-UVA (9 mW/cm2 for 10 minutes) are similar to standard "epi-off" corneal collagen crosslinking procedure in the treatment of progressive keratoconus.


TAp73 is required for spermatogenesis and the maintenance of male fertility.
The generation of viable sperm proceeds through a series of coordinated steps, including germ cell self-renewal, meiotic recombination, and terminal differentiation into functional spermatozoa. The p53 family of transcription factors, including p53, p63, and p73, are critical for many physiological processes, including female fertility, but little is known about their functions in spermatogenesis. Here, we report that deficiency of the TAp73 isoform, but not p53 or ΔNp73, results in male infertility because of severe impairment of spermatogenesis. Mice lacking TAp73 exhibited increased DNA damage and cell death in spermatogonia, disorganized apical ectoplasmic specialization, malformed spermatids, and marked hyperspermia. We demonstrated that TAp73 regulates the mRNA levels of crucial genes involved in germ stem/progenitor cells (CDKN2B), spermatid maturation/spermiogenesis (metalloproteinase and serine proteinase inhibitors), and steroidogenesis (CYP21A2 and progesterone receptor). These alterations of testicular histology and gene expression patterns were specific to TAp73 null mice and not features of mice lacking p53. Our work provides previously unidentified in vivo evidence that TAp73 has a unique role in spermatogenesis that ensures the maintenance of mitotic cells and normal spermiogenesis. These results may have implications for the diagnosis and management of human male infertility.


Use of pentapeptide-insertion scanning mutagenesis for functional mapping of the plum pox virus helper component proteinase suppressor of gene silencing.
Helper component proteinase (HC-Pro) of Plum pox virus is a multifunctional potyvirus protein that has been examined intensively. In addition to its involvement in aphid transmission, genome amplification and long-distance movement, it is also one of the better-studied plant virus suppressors of RNA silencing. The first systematic analysis using pentapeptide-insertion scanning mutagenesis of the silencing suppression function of a potyvirus HC-Pro is presented here. Sixty-three in-frame insertion mutants, each containing five extra amino acids inserted randomly within the HC-Pro protein, were analysed for their ability to suppress transgene-induced RNA silencing using Agrobacterium infiltration in transgenic Nicotiana benthamiana plants expressing green fluorescent protein. A functional map was obtained, consisting of clearly defined regions with different classes of silencing-suppression activity (wild-type, restricted and disabled). This map confirmed that the N-terminal part of the protein, which is indispensable for aphid transmission, is dispensable for silencing suppression and supports the involvement of the central region in silencing suppression, in addition to its role in maintenance of genome amplification and synergism with other viruses. Moreover, evidence is provided that the C-terminal part of the protein, previously known to be necessary mainly for proteolytic activity, also participates in silencing suppression. Pentapeptide-insertion scanning mutagenesis has been shown to be a fast and powerful tool to functionally characterize plant virus proteins.


Dietary magnesium deficiency alters gut microbiota and leads to depressive-like behaviour.
Gut microbiota (GM) has previously been associated with alterations in rodent behaviour, and since the GM is affected by the diet, the composition of the diet may be an important factor contributing to behavioural changes. Interestingly, a magnesium restricted diet has been shown to induce anxiety and depressive-like behaviour in humans and rodents, and it could be suggested that magnesium deficiency may mediate the effects through an altered GM. The present study therefore fed C57BL/6 mice with a standard diet or a magnesium deficient diet (MgD) for 6 weeks, followed by behavioural testing in the forced swim test (FST) to evaluate depressive-like behaviour. An intraperitoneal glucose tolerance test (GTT) was performed 2 day after the FST to assess metabolic alterations. Neuroinflammatory markers were analysed from hippocampus. GM composition was analysed and correlated to the behaviour and hippocampal markers. It was found that mice exposed to MgD for 6 weeks were more immobile than control mice in the FST, suggesting an increased depressive-like behaviour. No significant difference was detected in the GTT. GM composition correlated positively with the behaviour of undisturbed C57BL/6 mice, feeding MgD diet altered the microbial composition. The altered GM correlated positively to the hippocampal interleukin-6. In conclusion, we hypothesise that imbalances of the microbiota-gut-brain axis induced by consuming a MgD diet, contributes to the development of depressive-like behaviour.


Benzothiophenes as Potent Analgesics Against Neuropathic Pain.
Neuropathic pain arises because of neuronal injury. Unlike inflammatory pain which can be managed by classical nonsteroid anti-inflammatory drugs (NSAIDs), neuropathic pain is difficult to treat. The classical NSAIDs work through inhibition of cyclooxygenase 2 (COX2) enzyme. However, COX2 inhibitors are insufficient to treat neuropathic pain. Hence, it becomes important to explore for novel molecules acting through cell surface molecules like ion channels, for the treatment of neuropathic pain. We investigated multiple bromobenzothiophene carboxamides for their efficacy against neuropathic pain. Interestingly, AS6 was found to be very effective in treating neuropathic pain through inhibition of Kv4.3 ion channel. AS6 also reduced the COX2 overexpression associated with neuropathic pain. These results as well as results from our previous study indicate that AS6 can be a potent antinociceptive agent against both inflammatory and neuropathic pain.


Extramedullary relapse of AML with t(9;11)(p22;q23) associated with clonal evolution from trisomy 8 into tetrasomy 8.
This report describes a patient with extramedullary relapse of acute myeloid leukemia (AML) without involving bone marrow. A 57-year-old man was diagnosed as having acute monoblastic leukemia with t(9;11)(p22;q23) and trisomy 8. Ten months after achieving complete response (CR) with chemotherapy, masses developed in his left forearm and in the back of his thigh, preceded by enigmatic peripheral neurological symptoms. Aspiration from the forearm showed leukemic relapse, and fluorescence in situ hybridization (FISH) revealed that the majority of the cells had 11q23 anomaly and tetrasomy 8. Bone marrow or meningeal relapse was not observed. To our knowledge, this is the first case report of clonal evolution associated with the development of myeloid sarcoma as a relapse in AML.


Dimensions of pure chronic fatigue: psychophysical, cognitive and biological correlates in the chronic fatigue syndrome.
To investigate associated dimensions of fatigue regarding cognitive impairment, psychomotor performances, muscular effort power and circulating cytokine levels and their relations to symptom intensity in a sample of pure chronic fatigue syndrome (CFS) patients without overlapping objective sleepiness or sleep disorders. 16 CFS patients were compared to 14 matched controls. We assessed structured symptom-scales, polysomnography, multiple sleep latency tests, attention (Zazzo-Cancellation ZCT, digit-symbol-substitution DSST), psychomotor vigilance and speed (PVT, finger tapping test, FTT), dynamometer handgrip force (tonic and phasic trials) and circulating cytokines (IFN-γ, IL-1b, IL-6, IL-8, IL-10, TNF-α). In addition to fatigue, CFS patients presented with higher affective symptom intensity and worse perceived sleep quality. Polysomnography showed more slow-wave sleep and microarousals in CFS but similar sleep time, efficiency and light-sleep durations than controls. Patients presented with impaired attention (DSST, ZCT), slower reaction times (PVT) but not with lower hit rates (FTT). Notwithstanding lower grip strength during tonic and phasic trials, CFS also presented with higher fatigability during phasic trials. Cytokine levels were increased for IL-1b, IL-8, IL-10 and TNF-α and fatigue intensity was correlated to grip strength and IL-8. In contrast to sleepiness, chronic fatigue is a more complex phenomenon that cannot be reduced to one single measured dimension (i.e., sleep propensity). Showing its relations to different measurements, our study reflects this multidimensionality, in a psychosomatic disorder such as CFS. To obtain objective information, routine assessments of fatigue should rule out sleepiness, combine aspects of mental and physical fatigue and focus on fatigability.


Potential of spectral ratio indices derived from hyperspectral LiDAR and laser-induced chlorophyll fluorescence spectra on estimating rice leaf nitrogen contents.
Accurate estimation of leaf nitrogen contents (LNCs) is essential for nutrition management in monitoring crop growth status. The aim of this study was to compare the potential of hyperspectral LiDAR (HSL) and laser-induced chlorophyll fluorescence (LIF) data in accurately predicting rice LNC. First of all, the intensity values of HSL at 694 and 742 nm and LIF at ~685 and ~740 nm were selected as the characteristic variables to analyze rice LNC using data collected in 2014 and 2015, respectively. Second, spectral indices derived from HSL (only) and LIF (only) were utilized to estimate LNC of rice, respectively. Third, a combined ratio indices (the ratio indices of reflectance to fluorescence and NDVI-based indices at the above four wavelengths) was developed and evaluated in estimating rice LNC. The statistical method of linking these spectral indices to rice LNC was the artificial neural network, which was to obtain the optimum performance in LNC estimation of rice. The results demonstrated that the combined ratio indices, especially the ratio of reflectance to fluorescence at ~740 nm, showed a moderate relationship with rice LNC (R2 = 0.736, 0.704, and 0.713 for the 2014 first experiment, 2014 second experiment, and 2015 experiment, respectively).


Inner composition alignment for inferring directed networks from short time series.
Identifying causal links (couplings) is a fundamental problem that facilitates the understanding of emerging structures in complex networks. We propose and analyze inner composition alignment-a novel, permutation-based asymmetric association measure to detect regulatory links from very short time series, currently applied to gene expression. The measure can be used to infer the direction of couplings, detect indirect (superfluous) links, and account for autoregulation. Applications to the gene regulatory network of E. coli are presented.


Strength of tissue attachment to composite and ePTFE grafts after ventral hernia repair.
We sought to measure the strength of tissue attachment to mesh after laparoscopic ventral hernia repair in a porcine model. Twelve swine had two 10-cm x 16-cm sheets of ePTFE and polypropylene/ePTFE composite mesh fixated to the abdominal wall laparoscopically. Animals were euthanized at 2, 4, 6, and 12 weeks. The strength of tissue ingrowth was measured using a lap-shear method. Data are reported as mean force in pounds. Average surface area of adhesions to percentage of surface area was not statistically significant between the composite and ePTFE materials. For the composite material, there was a 98.7% posterior probability that the force required at 2 weeks was less than that required at 12 weeks. There was no difference in graft-abdominal wall interface strength between week 2 and week 12 for ePTFE material. Both prosthetics achieved the majority of their strength from tissue ingrowth by 2 weeks, but the composite prosthesis continued to gain strength while the strength of the ePTFE plateaued. Composite mesh demonstrated a statistically significant increase in strength between the lap-shear force, whereas no statistically significant difference occurred in the ePTFE graft. For the composite material, there was complete cellular infiltration through the entire thickness of polypropylene (approximately 500 microm) to the ePTFE layer at 2 weeks. At 2 weeks for ePTFE, the cells did not penetrate into the graft on the visceral side. On the abdominal wall side, the grooves filled with tissue, but no cellular penetration into the ePTFE occurred. No histological difference existed in cellularity. This study demonstrates that the strength of tissue ingrowth is significantly higher (P<0.05) for the composite grafts relative to the ePTFE grafts at each time point. Approximately 74% of tissue ingrowth and strength occurs by 2 weeks postoperatively for the composite prosthesis. The ePTFE graft tissue strength peaked and plateaued by 2 weeks. This may have clinical implications for human ventral hernia repair by partly addressing the issue of graft fixation to the abdominal wall during laparoscopic ventral hernia repair.


Wherever I may roam: Processes of self-esteem development from adolescence to emerging adulthood in the context of international student exchange.
Previous studies on self-esteem development show substantial changes as well as interindividual differences in change from adolescence to young adulthood. However, the processes underlying these developmental trajectories are still not well understood. The aim of the present study was to shed light on the macro- and microprocesses of self-esteem development. We investigated a sample of 876 German high school students (M = 16.0 years at Time 1) participating in an international exchange year. Exchange students provided 3 waves of trait self-esteem data (shortly before they departed, immediately after return, and 1 year later), as well as 9 monthly state measures of self-esteem and social inclusion during their stay abroad. In addition, a control group of high school students who stayed in Germany (N = 714) provided 2 waves of trait self-esteem data. From a macroperspective, results showed an effect of student exchange on trait self-esteem development: Exchange students showed a steeper mean-level increase and a lower rank-order stability compared with control students. Zooming in on the microprocesses underlying these developmental patterns, we found trait changes in exchange students to be mediated by state changes in self-esteem during their exchange. These fluctuations in state self-esteem were found to be predicted by feelings of social inclusion in the host country, and vice versa, providing support for both sociometer and self-broadcasting perspectives on self-esteem dynamics. In sum, our findings emphasize the importance of incorporating a microanalytical approach when investigating self-esteem development by showing that the environment triggers changes in this relatively stable personality trait through changes in states.


Pseudomonas putida response to cadmium: changes in membrane and cytosolic proteomes.
Pseudomonas putida is a saprophytic bacterium with remarkable environmental adaptability and the capacity to tolerate high concentrations of heavy metals. The strain P. putida-Cd001 was isolated from soil contaminated with Cd, Zn and Pb. Membrane-associated and cytosolic proteomes were analyzed to identify proteins whose expression was modulated in response to 250 μM CdSO4. We identified 44 protein spots in the membrane and 21 in the cytosolic fraction differentially expressed in Cd-treated samples compared to untreated controls. Outer membrane porins from the OprD and OprI families were less abundant in bacteria exposed to Cd, whereas those from the OprF and OprL, OprH and OprB families were more abundant, reflecting the increased need to acquire energy sources, the need to maintain membrane integrity and the process of adaptation. Components of the efflux system, such as the CzcB subunit of the CBA system, were also induced by Cd. Analysis of the cytosolic proteome revealed that proteins involved in protein synthesis, degradation and folding were induced along with enzymes that combat oxidative stress, showing that the entire bacterial proteome is modulated by heavy metal exposure. This analysis provides new insights into the adaptation mechanisms used by P. putida-Cd001 to survive in Cd-polluted environments.


Estradiol induces diurnal shifts in GABA transmission to gonadotropin-releasing hormone neurons to provide a neural signal for ovulation.
Ovulation is initiated by a surge of gonadotropin-releasing hormone (GnRH) secretion by the brain. GnRH is normally under negative feedback control by ovarian steroids. During sustained exposure to estradiol in the late follicular phase of the reproductive cycle, however, the feedback action of this steroid switches to positive, inducing the surge. Here, we used an established ovariectomized, estradiol-treated (OVX+E) mouse model exhibiting daily surges to investigate the neurobiological mechanisms underlying this switch. Specifically, we examined changes in GABA transmission to GnRH neurons, which can be excited by GABA(A) receptor activation. Spontaneous GABAergic postsynaptic currents (PSCs) were recorded in GnRH neurons from OVX+E and OVX mice in coronal and sagittal slices. There were no diurnal changes in PSC frequency in cells from OVX mice in either slice orientation. In OVX+E cells in both orientations, PSC frequency was low during negative feedback but increased at surge onset. During the surge peak, this increase subsided in coronal slices but persisted in sagittal slices. Comparison of PSCs before and during tetrodotoxin (TTX) treatment showed TTX decreased PSC frequency in OVX+E cells in sagittal slices, but not coronal slices. This indicates estradiol acts on multiple GABAergic afferent populations to increase transmission through both activity-dependent and -independent mechanisms. Estradiol also increased PSC amplitude during the surge. Estradiol and the diurnal cycle thus interact to induce shifts in both GABA transmission and postsynaptic response that would produce appropriate changes in GnRH neuron firing activity and hormone release.


Whole mitogenome sequencing uncovers a relation between mitochondrial heteroplasmy and leprosy severity.
In recent years, the mitochondria/immune system interaction has been proposed, so that variants of mitochondrial genome and levels of heteroplasmy might deregulate important metabolic processes in fighting infections, such as leprosy. We sequenced the whole mitochondrial genome to investigate variants and heteroplasmy levels, considering patients with different clinical forms of leprosy and household contacts. After sequencing, a specific pipeline was used for preparation and bioinformatics analysis to select heteroplasmic variants. We found 116 variants in at least two of the subtypes of the case group (Borderline Tuberculoid, Borderline Lepromatous, Lepromatous), suggesting a possible clinical significance to these variants. Notably, 15 variants were exclusively found in these three clinical forms, of which five variants stand out for being missense (m.3791T > C in MT-ND1, m.5317C > A in MT-ND2, m.8545G > A in MT-ATP8, m.9044T > C in MT-ATP6 and m.15837T > C in MT-CYB). In addition, we found 26 variants shared only by leprosy poles, of which two are characterized as missense (m.4248T > C in MT-ND1 and m.8027G > A in MT-CO2). We found a significant number of variants and heteroplasmy levels in the leprosy patients from our cohort, as well as six genes that may influence leprosy susceptibility, suggesting for the first time that the mitogenome might be involved with the leprosy process, distinction of clinical forms and severity. Thus, future studies are needed to help understand the genetic consequences of these variants.


Effectiveness of a multidisciplinary team approach to hip fracture management.
The purpose of the current study was to evaluate the effectiveness of a multidisciplinary Hip Fracture Service in the treatment of hip fractures in elderly patients. Baseline information and hospital outcomes were compared for 510 patients over the age of 65 with hip fracture treated before and after the institution of the Hip Fracture Service. Data included basic demographic data, admission laboratory results, surgical information, number of comorbidities, mortality, medical complications, discharge information, time to surgery, and length of stay in hospital. The demographics of the two groups of patients were similar. Patients treated as part of the Hip Fracture Service had fewer medical complications (36% vs. 51%), more often had surgery within 24 hours (63% vs. 35%), and had shorter hospital stays (mean, 5.7 days vs. 8.1 days) than patients treated before the Hip Fracture Service. These findings provide the rationale for a prospective, randomized trial of the service.


The improved sensitive detection of C-reactive protein based on the chemiluminescence immunoassay by employing monodispersed PAA-Au/Fe3O4 nanoparticles and zwitterionic glycerophosphoryl choline.
Nanocomposites are emerging complex nanoscale materials composed of different phases that have received considerable attention as a result of their widespread impact on a myriad of fields. Having said this, it is important to recognize that poor colloidal stabilities in physiological media and non-specific interactions with biology are key limitations prohibiting their successful translation into the development of practical technologies with real impact. Herein, PAA coated Au/Fe3O4 composite magnetic nanoparticles (NPs) with monodispersity and high biostability were engineered for the development of a highly sensitive C-reactive protein (CRP) chemiluminescence immunoassay (CLIA), based on the horseradish peroxidase (HRP)-luminol-H2O2 system. In addition, zwitterionic glycerophosphoryl choline (GPC) was employed as a co-blocking agent to reduce the nonspecific adsorption. This assay reports a limit of detection (LOD) of 2.5 pM with high reproducibility verified clinically and demonstrating improved progress towards translation into clinics.


Elevated CO2 causes changes in the photosynthetic apparatus of a toxic cyanobacterium, Cylindrospermopsis raciborskii.
We studied the physiological acclimation of growth, photosynthesis and CO2-concentrating mechanism (CCM) in Cylindrospermopsis raciborskii exposed to low (present day; L-CO2) and high (1300ppm; H-CO2) pCO2. Results showed that under H-CO2 the cell specific division rate (μc) was higher and the CO2- and light-saturated photosynthetic rates (Vmax and Pmax) doubled. The cells' photosynthetic affinity for CO2 (K0.5CO2) was halved compared to L-CO2 cultures. However, no significant differences were found in dark respiration rates (Rd), pigment composition and light harvesting efficiency (α). In H-CO2 cells, non-photochemical quenching (NPQ), associated with state transitions of the electron transport chain (ETC), was negligible. Simultaneously, a reorganisation of PSII features including antenna connectivity (JconPSIIα), heterogeneity (PSIIα/β) and effective absorption cross sectional area (σPSIIα/β) was observed. In relation to different activities of the CCM, our findings suggest that for cells grown under H-CO2: (1) there is down-regulation of CCM activity; (2) the ability of cells to use the harvested light energy is altered; (3) the occurrence of state transitions is likely to be associated with changes of electron flow (cyclic vs linear) through the ETC; (4) changes in PSII characteristics are important in regulating state transitions.


RAB35 depletion affects spindle formation and actin-based spindle migration in mouse oocyte meiosis.
Mammalian oocyte maturation involves a unique asymmetric cell division, in which meiotic spindle formation and actin filament-mediated spindle migration to the oocyte cortex are key processes. Here, we report that the vesicle trafficking regulator, RAB35 GTPase, is involved in regulating cytoskeleton dynamics in mouse oocytes. RAB35 GTPase mainly accumulated at the meiotic spindle periphery and cortex during oocyte meiosis. Depletion of RAB35 by morpholino microinjection led to aberrant polar body extrusion and asymmetric division defects in almost half the treated oocytes. We also found that RAB35 affected SIRT2 and αTAT for tubulin acetylation, which further modulated microtubule stability and meiotic spindle formation. Additionally, we found that RAB35 associated with RHOA in oocytes and modulated the ROCK-cofilin pathway for actin assembly, which further facilitated spindle migration for oocyte asymmetric division. Importantly, microinjection of Myc-Rab35 cRNA into RAB35-depleted oocytes could significantly rescue these defects. In summary, our results suggest that RAB35 GTPase has multiple roles in spindle stability and actin-mediated spindle migration in mouse oocyte meiosis.


Contemporary management of concomitant carotid and coronary artery disease.
The best approach to the management of concomitant severe carotid and coronary artery disease remains unanswered. The American College of Cardiology/American Heart Association (ACC/AHA) guidelines recommend carotid endarterectomy (CEA) in asymptomatic carotid stenosis of ≥ 80% either prior to or combined with coronary artery bypass surgery (CABG). Currently, there is no consensus as to which surgical approach is superior. More recently, carotid artery stenting (CAS) prior to CABG is emerging as an alternative option with promising results in asymptomatic patients considered 'high risk' for CEA. A <3% composite event rate has been set as a benchmark for isolated CAS or CEA in asymptomatic patients by the ACC/AHA; however, most CEA or CAS studies in patients requiring concomitant CABG have shown event rates ranging from 10-12%. This review examines the available data on carotid revascularisation in relation to CABG surgery to aid in the risk-benefit decision analysis in this controversial area.


Elevation of soluble E-selectin levels following gestational diabetes is restricted to women with persistent abnormalities of glucose regulation.
Increased levels of the soluble adhesion molecule sE-selectin have been reported in normoglycaemic women with previous gestational diabetes but not in other groups at increased risk of future type 2 diabetes. To explore the basis for these discrepant findings, we studied the relationship between sE-selectin and glucose regulation in a large group of women with previous gestational diabetes. Comparison of sE-selectin levels between a study cohort ascertained on the basis of recent gestational diabetes and suitable control subjects. One hundred and forty women with recent gestational diabetes (104 European, 20 South Asian and 16 Afro-Caribbean) and 125 normoglycaemic control women (90 European, 19 South Asian and 16 Afro-Caribbean). sE-selectin, fasting lipids, insulin and current glucose regulation status. There was no overall difference in sE-selectin levels between women with a history of gestational diabetes and control women among the 3 ethnic groups. European post-GDM women with abnormal glucose regulation postpartum (n = 30) had higher sE-selectin than control women (67 (54-91) ng/ml vs. 57 (43-75) ng/ml, P = 0.049). There was no difference in sE-selectin between normoglycaemic European women with a history of gestational diabetes (n = 74) and control women, even though the former displayed metabolic abnormalities predictive of diabetes. In those European post-GDM women with normal glucose regulation postpartum, sE-selectin levels were negatively correlated with time since delivery (r = -0.25, P = 0.04), suggesting that the previously described elevation of sE-selectin following GDM pregnancies slowly resolves postpartum. There was no correlation between sE-selectin levels and features of insulin resistance. In contrast to previous findings, this larger study does not support a role for sE-selectin as a marker, independent of prevailing glucose levels, of early metabolic abnormalities or future diabetes risk in women with previous gestational diabetes.


Rat precision-cut intestinal slices to study P-gp activity and the potency of its inhibitors ex vivo.
Rat Precision-Cut Intestinal Slices (PCIS) were evaluated as ex vivo model to study the regional gradient of P-gp activity, and to investigate whether the rank order of inhibitory potency of P-gp inhibitors can be correctly reproduced in this model with more accurate IC50 values than with current in vitro models. PCIS were prepared from small intestine (duodenum, jejunum, ileum) and colon. Rhodamine 123 (R123) was used as P-gp substrate, while verapamil, cyclosporine A, quinidine, ketoconazole, PSC833 and CP100356 were employed as P-gp inhibitors. Increase in tissue accumulation of R123 in the presence of the inhibitors was considered as an indication of the inhibitory effect. The P-gp inhibitors increased the tissue accumulation of R123 in a concentration dependent manner. Fluorescence microscopy elucidated that this increase occurred predominantly in the enterocytes. The rank order of the corresponding IC50 values agreed well with reported values from cell lines expressing rat P-gp. The activity of and inhibitory effects on P-gp were significantly higher in ileum compared to the other regions. These data suggest that rat PCIS are a reliable ex vivo model to study the activity of intestinal P-gp and the inhibitory effect of drugs. PCIS have potential as ex vivo model for the prediction of transporter-mediated drug-drug interactions.


Validation of Prediction Models for Critical Care Outcomes Using Natural Language Processing of Electronic Health Record Data.
Accurate prediction of outcomes among patients in intensive care units (ICUs) is important for clinical research and monitoring care quality. Most existing prediction models do not take full advantage of the electronic health record, using only the single worst value of laboratory tests and vital signs and largely ignoring information present in free-text notes. Whether capturing more of the available data and applying machine learning and natural language processing (NLP) can improve and automate the prediction of outcomes among patients in the ICU remains unknown. To evaluate the change in power for a mortality prediction model among patients in the ICU achieved by incorporating measures of clinical trajectory together with NLP of clinical text and to assess the generalizability of this approach. This retrospective cohort study included 101 196 patients with a first-time admission to the ICU and a length of stay of at least 4 hours. Twenty ICUs at 2 academic medical centers (University of California, San Francisco [UCSF], and Beth Israel Deaconess Medical Center [BIDMC], Boston, Massachusetts) and 1 community hospital (Mills-Peninsula Medical Center [MPMC], Burlingame, California) contributed data from January 1, 2001, through June 1, 2017. Data were analyzed from July 1, 2017, through August 1, 2018. In-hospital mortality and model discrimination as assessed by the area under the receiver operating characteristic curve (AUC) and model calibration as assessed by the modified Hosmer-Lemeshow statistic. Among 101 196 patients included in the analysis, 51.3% (n = 51 899) were male, with a mean (SD) age of 61.3 (17.1) years; their in-hospital mortality rate was 10.4% (n = 10 505). A baseline model using only the highest and lowest observed values for each laboratory test result or vital sign achieved a cross-validated AUC of 0.831 (95% CI, 0.830-0.832). In contrast, that model augmented with measures of clinical trajectory achieved an AUC of 0.899 (95% CI, 0.896-0.902; P < .001 for AUC difference). Further augmenting this model with NLP-derived terms associated with mortality further increased the AUC to 0.922 (95% CI, 0.916-0.924; P < .001). These NLP-derived terms were associated with improved model performance even when applied across sites (AUC difference for UCSF: 0.077 to 0.021; AUC difference for MPMC: 0.071 to 0.051; AUC difference for BIDMC: 0.035 to 0.043; P < .001) when augmenting with NLP at each site. Intensive care unit mortality prediction models incorporating measures of clinical trajectory and NLP-derived terms yielded excellent predictive performance and generalized well in this sample of hospitals. The role of these automated algorithms, particularly those using unstructured data from notes and other sources, in clinical research and quality improvement seems to merit additional investigation.


Trends in metals in urban and reference lake sediments across the United States, 1970 to 2001.
Trends in metals concentrations in sediment cores from 35 reservoirs and lakes in urban and reference settings were analyzed to determine the effects of three decades of legislation, regulation, and changing demographics and industrial practices in the United States on concentrations of metals in the environment. Decreasing trends outnumber increasing trends for all seven metals analyzed (Cd, Cr, Cu, Pb, Hg, Ni, and Zn). The most consistent trends are for Pb and Cr: For Pb, 83% of the lakes have decreasing trends and 6% have increasing trends; for Cr, 54% of the lakes have decreasing trends and none have increasing trends. Mass accumulation rates of metals in cores, adjusted for background concentrations, decrease from the 1970s to the 1990s, with median changes ranging from -46% (Pb) to -3% (Hg and Zn). The largest decreases are from lakes in dense urban watersheds where the overall metals contamination in recently deposited sediments has decreased to one-half its 1970s median value. However, anthropogenic mass accumulation rates in dense urban lakes remain elevated over those in lakes in undeveloped watersheds, in some cases by as much as two orders of magnitude (Cr, Cu, and Zn), indicating that urban fluvial source signals can overwhelm those from regional atmospheric sources.


Feedback regulations of miR-21 and MAPKs via Pdcd4 and Spry1 are involved in arsenite-induced cell malignant transformation.
To establish the functions of miR-21 and the roles of two feedback regulation loops, miR-21-Spry1-ERK/NF-κB and miR-21-Pdcd4-JNK/c-Jun, in arsenite-transformed human embryo lung fibroblast (HELF) cells. For arsenite-transformed HELF cells, apoptosis, clonogenicity, and capacity for migration were determined by Hoechst staining, assessment of their capacity for anchorage-independent growth, and wound-healing, respectively, after blockage, with inhibitors or with siRNAs, of signal pathways for JNK/c-Jun or ERK/NF-κB. Decreases of miR-21 levels were determined with anti-miR-21, and the up-regulation of Pdcd4 and Spry1 was assessed in transfected cells; these cells were molecularly characterized by RT-PCR, qRT-PCR, Western blots, and immunofluorescence assays. MiR-21 was highly expressed in arsenite-transformed HELF cells and normal HELF cells acutely treated with arsenite, an effect that was concomitant with activation of JNK/c-Jun and ERK/NF-κB and down-regulation of Pdcd4 and Spry1 protein levels. However, there were no significant changes in mRNA levels for Pdcd4 and Spry1, which suggested that miR-21 regulates the expressions of Pdcd4 and Spry1 through translational repression. In arsenite-transformed HELF cells, blockages of JNK/c-Jun or ERK/NF-κB with inhibitors or with siRNAs prevented the increases of miR-21and the decreases of the protein levels but not the mRNA levels of Pdcd4 and Spry1. Down-regulation of miR-21 and up-regulations of Pdcd44 or Spry1 blocked the arsenite-induced activations of JNK/c-Jun or ERK/NF-κB, indicating that knockdown of miR-21 inhibits feedback of ERK activation and JNK activation via increases of Pdcd4 and Spry1 protein levels, respectively. Moreover, in arsenite-transformed HELF cells, inhibition of miR-21 promoted cell apoptosis, inhibited clonogenicity, and reduced migration. The results indicate that miR-21 is both a target and a regulator of ERK/NF-κB and JNK/c-Jun and the feedback regulations of miR-21 and MAPKs via Pdcd4 and Spry1, respectively, are involved in arsenite-induced malignant transformation of HELF cells.


The use of injectable alphaxalone as a single agent and in combination with ketamine, xylazine, and morphine in the Chilean rose tarantula, Grammostola rosea.
This study evaluated the use of the injectable anesthetic, alphaxalone, as a single agent and in combination with ketamine, xylazine, and morphine in the Chilean rose tarantula, Grammostola rosea. Between two and four animals were evaluated for each anesthetic protocol, and two unanesthetized animals were evaluated for comparative purposes. Anesthetic duration, depth, and quality were assessed by scoring responses to tactile and trichobothria stimulation, muscle tone, purposeful movement, righting response, and heart rate throughout each anesthetic event. Alphaxalone administered into the dorsal opisthosoma in the location of the heart at 200 mg/kg produced moderate anesthetic effect with a median duration of 28 min (n = 3; range 25-50). A combination of 200 mg/kg of alphaxalone and 20 mg/kg of ketamine induced a deep anesthetic state with a median anesthetic duration of 27 min (n = 4; range 16-42). The combination of 200 mg/kg of alphaxalone and 20 mg/kg of xylazine produced deep anesthesia with a median duration of 70 min (n = 4; range 37-207). Morphine administered at 5 mg/kg 30 min prior to injection with 200 mg/kg alphaxalone had anesthetic durations of 9 and 30 min (n = 2). Heartbeats could not be detected for periods of 7-27 min following anesthetic induction for the majority of animals receiving the alphaxalone/ketamine and alphaxalone/xylazine anesthetic combinations. No mortality was associated with any of the anesthetic protocols used; however, ambient temperature and ecdysis were identified as important factors that may alter response to anesthetics in these animals.


A Di-leucine signal in the ubiquitin moiety. Possible involvement in ubiquitination-mediated endocytosis.
Some plasma membrane receptors in yeast are known to be internalized and degraded in lysosomes upon ligand-dependent ubiquitination. However, the role of ubiquitination in endocytosis and lysosomal degradation in higher eukaryotes has been controversial. In order to directly assess this question, we investigated the fate of chimeric molecules in which ubiquitin moiety was fused in-frame to the cytoplasmic region of membrane proteins. The chimeric proteins with the wild-type ubiquitin were endocytosed and delivered to lysosomes efficiently. Mutant ubiquitin with lysine-to-arginine substitution could still mediate endocytosis, suggesting that polyubiquitination is not required for the endocytosis. We next searched for the existence of an endocytosis signal(s) in the ubiquitin moiety, and identified a di-leucine signal, Leu(43)-Ile(44). The Leu(43)-Ile(44) sequence mediated endocytosis and lysosomal sorting in a Leu(43)-dependent manner. These results suggest that the di-leucine signal in ubiquitin can be involved in ubiquitination-mediated endocytosis and lysosomal targeting of membrane proteins.


A bacterial toxin that controls cell cycle progression as a deoxyribonuclease I-like protein.
Many bacterial pathogens encode a multisubunit toxin, termed cytolethal distending toxin (CDT), that induces cell cycle arrest, cytoplasm distention, and, eventually, chromatin fragmentation and cell death. In one such pathogen, Campylobacter jejuni, one of the subunits of this toxin, CdtB, was shown to exhibit features of type I deoxyribonucleases. Transient expression of this subunit in cultured cells caused marked chromatin disruption. Microinjection of low amounts of CdtB induced cytoplasmic distention and cell cycle arrest. CdtB mutants with substitutions in residues equivalent to those required for catalysis or magnesium binding in type I deoxyribonucleases did not cause chromatin disruption. CDT holotoxin containing these mutant forms of CdtB did not induce morphological changes or cell cycle arrest.


TCR alpha-chain repertoire in pTalpha-deficient mice is diverse and developmentally regulated: implications for pre-TCR functions and TCRA gene rearrangement.
Pre-TCR expression on developing thymocytes allows cells with productive TCRB gene rearrangements to further differentiate. In wild-type mice, most TCRA gene rearrangements are initiated after pre-TCR expression. However, in pTalpha-deficient mice, a substantial number of alphabeta+ thymocytes are still produced, in part because early TCR alpha-chain expression can rescue immature thymocytes from cell death. In this study, the nature of these TCR alpha-chains, produced and expressed in the absence of pre-TCR expression, have been analyzed. We show, by FACS analysis and sequencing of rearranged transcripts, that the TCRA repertoire is diverse in pTalpha-/- mice and that the developmental regulation of AJ segment use is maintained, yet slightly delayed around birth when compared with wild-type mice. We also found that T cell differentiation is more affected by pTalpha inactivation during late gestation than later in life. These data suggest that the pre-TCR is not functionally required for the initiation and regulation of TCRA gene rearrangement and that fetal thymocytes are more dependent than adult cells on pTalpha-derived signals for their differentiation.


Effects of prior treatment with simvastatin on skeletal muscle structure and mitochondrial enzyme activities during early phases of sepsis.
We investigated the effects of the early phase of sepsis and prior treatment of Simvastatin on muscle structure and mitochondrial enzymes treated with lipopolysaccharide (LPS) in rats. We divided rats into control, LPS, simvastatin, simvastatin + LPS groups. Mitochondrial citrate synthase, complex I, II, I + III, II + III, cytochrome c oxidase (COX) activities were measured. Muscle tissue was stained using modified Gomori trichrome (MGT), succinic dehydrogenase (SDH) and cytochrome oxidase (COX). In all treated groups, complex I and citrate synthase activities were higher than in the controls. In the control and LPS groups, COX activity was increased when compared with simvastatins'. Complex II, II-III activities were higher in the LPS group than in the control group. Complex I-III activities were higher in the Simvastatin and Simvastatin + LPS groups than in the control and LPS groups (P < 0.05). Myopathic changes with LPS group were observed in MGT stained sections. Our findings showed improvements in the alterations of enzyme activities and muscle myofibrils after treating rats with LPS that had received a prior dose of simvastatin.


HIV in Africa (Communication arising): chemokine-receptor genes and AIDS risk.
Schliekelman et al. have provided a model to quantify the speed at which HIV-resistance haplotypes can become enriched in a susceptible population through a delay in the onset of AIDS, permitting greater lifetime reproduction and the selection of AIDS-delaying haplotypes. But we question their conclusion that there could be a rapid evolution of resistance to AIDS onset in some African populations if the current HIV epidemic persists, as this depends on an untested assumption: that variant forms of the chemokine-receptor-5 (CCR5) gene impart selective advantages or disadvantages in Africa that are comparable to those reported for African Americans. Here we test this premise in a large Ugandan population, and find that CCR5 variants are not associated with HIV/AIDS disease risk in Africa--the origin and centre of the current AIDS pandemic. This gene may therefore not be subject to rapid evolutionary change as a result of the HIV epidemic in Africa.


[The psychiatric hospital: from yesterday to tomorrow].
A study carried out in a psychiatric hospital in the Montreal region reveals that in spite of deinstitutionalization, long-term stay remains an important factor in the use of beds. For some patients the hospital remains a permanent home, either from their first admission or from the time they are institutionalized after multiple admissions. For both in- and out-patients, return to the hospital and long-term care are almost inevitable. Generally speaking, the services of the hospital are used by the same long-term patients, and this to the extent that the hospital cannot provide services to other establishments or play its second-line role. This is frustrating for other institutions in the network, even though they accept the inevitability of the situation given the lack of adequate community resources for these patients. One of the primary functions of a psychiatric hospital also seems to be to fill in the gaps in the service network. In this regard, its expertise in the field of intervention in chronic psychiatric patients must not be overlooked. Given the current situation, the hospital's responsibility in regard to second-line services cannot be clarified until it has been determined just how much of the responsibility for care of psychiatric patients can be assumed by the community itself.


Neutralization epitopes of human rhinovirus type 2.
Fourteen neutralizing monoclonal antibodies recognizing human rhinovirus (HRV) type 2 have been used to select a total of 51 virus escape mutants. Cross-resistance analysis of the mutants, together with RNA sequencing and identification of amino acid substitutions, have revealed three neutralization sites on the virus surface. Two of these appear to correspond to the NIm-IA and NIm-II sites described for HRV-type 14, although there are also substantial differences. The third site has not been described previously.


Schmallenberg virus detection in bovine semen after experimental infection of bulls.
To study Schmallenberg virus (SBV) excretion in bovine semen after experimental infection, two bulls were inoculated subcutaneously with a SBV isolate (1 ml Vero cell culture 106 TCID50). After inoculation (at day 0), semen was collected daily from both animals for 21 days and samples were tested for SBV by qRT-PCR assay. At 24 days post-inoculation both animals were subjected to necropsy and the genital organs and lymph nodes draining these organs were also tested for SBV RNA (qRT-PCR). After SBV infection both animals in the study showed viraemia (qRT-PCR) with fever and diarrhoea. SBV RNA could be detected in semen from both animals. The highest SBV RNA concentrations in semen were found in the first week (days 4-7 post-inoculation) but concentrations were relatively low (Ct values 30-39). Viable SBV was only isolated from blood samples and not from semen or genital tissues.


Endocrine disruption potentials of organophosphate flame retardants and related mechanisms in H295R and MVLN cell lines and in zebrafish.
Organophosphate flame retardants (OPFRs) are frequently detected in environment and biota. However, knowledge on their potential toxicological effects is limited. Endocrine disrupting potentials of six OPFRs, i.e., tris-(2-chloroethyl) phosphate (TCEP), tris-(2-chloroisopropyl) phosphate (TCPP), tris-(1,3-dichloro-2-propyl) phosphate (TDCPP), tris-(2-butoxyethyl) phosphate (TBEP), triphenyl phosphate (TPP), and tricresyl phosphate (TCP), were investigated using human cell lines as well as zebrafish (Danio rerio). Sex hormone synthesis and steroidogenic gene transcriptions were measured using H295R cells. With MVLN cells, estrogen receptor binding activities of OPFRs were evaluated. In zebrafish, sex hormones and related gene transcriptions were determined for each sex after 14d exposure to OPFRs. All six OPFRs increased both 17β-estradiol (E2) and testosterone (T) concentrations in H295R cells. In addition, transcription of four major steroidogenic genes was up-regulated and that of two sulfotransferase genes was down-regulated. In MVLN cells, no OPFRs acted as estrogen receptor agonists, while TDCPP, TPP, and TCP acted as antagonists inhibiting binding of E2 to estrogen receptor. After 14d of zebrafish exposure, TCP, TDCPP, or TPP significantly increased plasma T and E2 concentrations, but did not change 11-ketotestosterone (11-KT) among female fish. Among males, both T and 11-KT decreased and E2 increased. In general, transcription of CYP17 and CYP19a genes was significantly up-regulated in both sexes, while vitellogenin (VTG) 1 gene was down- and up-regulated in female and male fish, respectively. The results of this study showed that OPFRs could alter sex hormone balance through several mechanisms including alterations of steroidogenesis or estrogen metabolism.


Melatonin inhibits the expression of vascular endothelial growth factor in pancreatic cancer cells.
To investigate the effects of melatonin on cellular proliferation and endogenous vascular endothelial growth factor (VEGF) expression in pancreatic carcinoma cells (PANC-1). PANC-1 cells were cultured for this study. The secreted VEGF concentration in the culture medium was determined using ELISA method, VEGF production in the tumor cells was detected by immunocytochemistry, and VEGF mRNA expression was determined by RT-PCR. Higher melatonin concentrations significantly inhibited cellular proliferation, with 1 mmol/L concentration exhibiting the highest inhibitory effect (P<0.01). VEGF concentrations in the cell culture supernatants and intra-cellules were all significantly reduced after melatonin (1 mmol/L) incubation (P<0.05). VEGF mRNA expression decreased markedly in a time-dependent manner during the observation period (P<0.05). High melatonin concentrations markedly inhibited the proliferation of pancreatic carcinoma cells. The endogenous VEGF expression was also suppressed by melatonin incubation.


Patients' perceptions with musculoskeletal disorders regarding their experience with healthcare providers and health services: an overview of reviews.
This overview of reviews aimed to identify (1) aspects of the patient experience when seeking care for musculoskeletal disorders from healthcare providers and the healthcare system, and (2) which mechanisms are used to measure aspects of the patient experience. Four databases were searched from inception to December 20th, 2019. Systematic or scoping reviews examining patient experience in seeking care for musculoskeletal from healthcare providers and the healthcare system were included. Independent authors screened and selected studies, extracted data, and assessed the methodological quality of the reviews. Patient experience concepts were compiled into five themes from a perspective of a) relational and b) functional aspects. A list of mechanisms used to capture the patient experience was also collected. Thirty reviews were included (18 systematic and 12 scoping reviews). Relational aspects were reported in 29 reviews and functional aspects in 25 reviews. For relational aspects, the most prevalent themes were "information needs" (education and explanation on diseases, symptoms, and self-management strategies) and "understanding patient expectations" (respect and empathy). For functional aspects, the most prevalent themes were patient's "physical and environmental needs," (cleanliness, safety, and accessibility of clinics), and "trusted expertise," (healthcare providers' competence and clinical skills to provide holistic care). Interviews were the most frequent mechanism identified to collect patient experience. Measuring patient experience provides direct insights about the patient's perspectives and may help to promote better patient-centered health services and increase the quality of care. Areas of improvement identified were interpersonal skills of healthcare providers and logistics of health delivery, which may lead to a more desirable patient-perceived experience and thus better overall healthcare outcomes. Systematic review registration: PROSPERO (CRD42019136500).


Outpatient treatment of microbial corneal ulcers.
Outpatient and inpatient management of culture-proved microbial corneal ulcers was evaluated. Twenty-six consecutive patients were reviewed. Twenty had been managed as outpatients, six as inpatients. All ulcers were treated with intensive topical antimicrobial therapy, and all steadily healed, as evidenced by epithelialization and resolution of infiltrate. None progressed to extreme stromal thinning, descemetocele, or perforation. Inpatients and outpatients did not differ in terms of healing time, although this can vary considerably for the same causative organism. Complete epithelialization took two to 56 days. Pseudomonas ulcers healed in seven to 42 days (mean, 22 days), which is similar to previously published reports of inpatient therapy. This study suggests that with careful patient selection and follow-up, infectious corneal ulcers can be successfully managed on an outpatient basis.


Identification of a synthetic peptide as part of a major neutralization epitope of respiratory syncytial virus.
A 7000 Mr cleavage fragment of the F1 subunit that carries the major neutralization epitope has been identified by chemical and enzymatic cleavage of the fusion protein of respiratory syncytial (RS) virus (Long strain) with an efficient RS virus-neutralizing monoclonal antibody. Based on the published mRNA-deduced sequence of the A2 strain, coupled to the hydropathicity profile and prediction of protein conformation, the neutralization epitope has tentatively been localized on the first third of the F1 protein N-terminal, probably in the region of amino acids 215Ser to 236Glu. Analysis of three peptides covering different portions of the 212Cys to 236Glu region of the F1 fusion protein identified a peptide (Cys X 216Asn to 236Glu) that reacted strongly with the neutralizing monoclonal antibody and that was efficient in blocking neutralization and in plaque-reducing assays, confirming that the neutralization epitope was localized in that region. Further analysis with two other synthetic peptides (212Cys to 222Glu and Cys X 221Ile to 236Glu) indicated that the dodecapeptide Ile-Glu-Phe-Gln-Lys-Asn-Asn-Arg-Leu-Leu-Glu mimicked either the whole or a major part of the neutralization epitope. This opens a promising avenue for the simple design of a synthetic peptide vaccine to control RS virus infection.


Relationship between acuity for gratings and for tumbling-E letters in peripheral vision.
Earlier studies have reported that grating resolution is sampling-limited in peripheral vision but that letter acuity is generally poorer than grating acuity. These results suggest that peripheral resolution of objects with rich Fourier spectra may be limited by some factor other than neural sampling. To examine this suggestion we formulated and tested the hypothesis that letter acuity in the periphery is sampling-limited, just as it is for extended and truncated gratings. We tested this hypothesis with improved methodology to avoid the confounding factors of target similarity, alphabet size, individual variation, peripheral refractive error, and stimulus size. Acuity was measured for an orientation-discrimination task (horizontal versus vertical) for a three-bar resolution target and for a block-E letter in which all strokes have the same length. We confirmed previous reports in the literature that acuity for these targets is worse than for extended sinusoidal gratings. To account for these results quantitatively, we used difference-spectrum analysis to identify those frequency components of the targets that might form a basis for performing the visual discrimination task. We find that discrimination performance for the three-bar targets and the block-E letters can be accounted for by a sampling-limited model, provided that the limited number of cycles that are present in the characteristic frequency of the stimulus is taken into account. Quantitative differences in acuity for discriminating other letter pairs (e.g., right versus left letters E or characters with short central strokes) could not be attributed to undersampling of either the characteristic frequency or the frequency of maximum energy in the difference spectrum. These results suggest additional tests of the sampling theory of visual resolution, which are the subject of a companion paper.


Extracting morphologies from third harmonic generation images of structurally normal human brain tissue.
The morphologies contained in 3D third harmonic generation (THG) images of human brain tissue can report on the pathological state of the tissue. However, the complexity of THG brain images makes the usage of modern image processing tools, especially those of image filtering, segmentation and validation, to extract this information challenging. We developed a salient edge-enhancing model of anisotropic diffusion for image filtering, based on higher order statistics. We split the intrinsic 3-phase segmentation problem into two 2-phase segmentation problems, each of which we solved with a dedicated model, active contour weighted by prior extreme. We applied the novel proposed algorithms to THG images of structurally normal ex-vivo human brain tissue, revealing key tissue components-brain cells, microvessels and neuropil, enabling statistical characterization of these components. Comprehensive comparison to manually delineated ground truth validated the proposed algorithms. Quantitative comparison to second harmonic generation/auto-fluorescence images, acquired simultaneously from the same tissue area, confirmed the correctness of the main THG features detected. The software and test datasets are available from the authors. email@example.com. Supplementary data are available at Bioinformatics online.


Knee mega-prosthesis in the management of complex knee fracture of the elderly : a case series and review of the literature.
The management of complicated distal femur fractures (DFF) of the elderly continues to pose a challenge. Knee mega-prosthesis are mostly used for Total knee arthroplasty revision and tumor resection surgery but they can be used for the treatment of complex knee fractures. The purpose of the present study is to examine the short- to mid- term outcomes of their use for complex DFF of the elderly. We retrospectively identified 4 patients with DFF AO33C3 on osteoporotic bone treated by total knee arthroplasty from September 2015 to October 2019. The average age at the time of the surgery was 79,5 years (range, 69 to 95 years). All patients were females and underwent a total knee replacement by one senior surgeon, with the OSS TM Orthopaedic Salvage System (Zimmer Biomet, Warsaw, Indiana, USA). Outcome measures included clinical outcome scores, radiological analyses, reoperation rate and complications. At an average follow-up of 2,3 years (range, 0,6 to 4,2 years), the average Western Ontario and McMaster Universities Osteoarthritis index (WOMAC) was 17,25 (range, 7 to 37), the average Oxford knee score was 35,25 (range, 25 to 41) and the average pain Numerical Rating Scale (NRS) was 0,5 (range, 0 to 1). 3 Patients had postoperative anemia but no implant related complications has been reported. Complex DFF of the elderly treated with mega knee arthroplasty exhibit good clinical outcomes scores. The patients should be selected carefully as the complication rate found in the literature remains high.


Orientational Order in Self-Assembled Nanocrystal Superlattices.
Self-assembly of nanocrystals into functional materials requires precise control over nanoparticle interactions in solution that are dominated by organic ligands that densely cover the surface of nanocrystals. Recent experiments have demonstrated that small truncated-octahedral nanocrystals can self-assemble into a range of superstructures with different translational and orientational order of nanocrystals. The origin of this structural diversity remains unclear. Here, we use molecular dynamics computer simulations to study the self-assembly of these nanocrystals over a broad range of ligand lengths and solvent conditions. Our model, which is based on a coarse-grained description of ligands and solvent effects, reproduces the experimentally observed superstructures, including recently observed superlattices with partial and short-ranged orientational alignment of nanocrystals. We show that small differences in nanoparticle shape, ligand length and coverage, and solvent conditions can lead to markedly different self-assembled superstructures due to subtle changes in the free energetics of ligand interactions. Our results rationalize the large variety of different reported superlattices self-assembled from seemingly similar particles and can serve as a guide for the targeted self-assembly of nanocrystal superstructures.


Limited brain metabolism changes differentiate between the progression and clearance of rabies virus.
Central nervous system (CNS) metabolic profiles were examined from rabies virus (RABV)-infected mice that were either mock-treated or received post-exposure treatment (PET) with a single dose of the live recombinant RABV vaccine TriGAS. CNS tissue harvested from mock-treated mice at middle and late stage infection revealed numerous changes in energy metabolites, neurotransmitters and stress hormones that correlated with replication levels of viral RNA. Although the large majority of these metabolic changes were completely absent in the brains of TriGAS-treated mice most likely due to the strong reduction in virus spread, TriGAS treatment resulted in the up-regulation of the expression of carnitine and several acylcarnitines, suggesting that these compounds are neuroprotective. The most striking change seen in mock-treated RABV-infected mice was a dramatic increase in brain and serum corticosterone levels, with the later becoming elevated before clinical signs or loss of body weight occurred. We speculate that the rise in corticosterone is part of a strategy of RABV to block the induction of immune responses that would otherwise interfere with its spread. In support of this concept, we show that pharmacological intervention to inhibit corticosterone biosynthesis, in the absence of vaccine treatment, significantly reduces the pathogenicity of RABV. Our results suggest that widespread metabolic changes, including hypothalamic-pituitary-adrenal axis activation, contribute to the pathogenesis of RABV and that preventing these alterations early in infection with PET or pharmacological blockade helps protect brain homeostasis, thereby reducing disease mortality.


Color Shift Failure Prediction for Phosphor-Converted White LEDs by Modeling Features of Spectral Power Distribution with a Nonlinear Filter Approach.
With the expanding application of light-emitting diodes (LEDs), the color quality of white LEDs has attracted much attention in several color-sensitive application fields, such as museum lighting, healthcare lighting and displays. Reliability concerns for white LEDs are changing from the luminous efficiency to color quality. However, most of the current available research on the reliability of LEDs is still focused on luminous flux depreciation rather than color shift failure. The spectral power distribution (SPD), defined as the radiant power distribution emitted by a light source at a range of visible wavelength, contains the most fundamental luminescence mechanisms of a light source. SPD is used as the quantitative inference of an LED's optical characteristics, including color coordinates that are widely used to represent the color shift process. Thus, to model the color shift failure of white LEDs during aging, this paper first extracts the features of an SPD, representing the characteristics of blue LED chips and phosphors, by multi-peak curve-fitting and modeling them with statistical functions. Then, because the shift processes of extracted features in aged LEDs are always nonlinear, a nonlinear state-space model is then developed to predict the color shift failure time within a self-adaptive particle filter framework. The results show that: (1) the failure mechanisms of LEDs can be identified by analyzing the extracted features of SPD with statistical curve-fitting and (2) the developed method can dynamically and accurately predict the color coordinates, correlated color temperatures (CCTs), and color rendering indexes (CRIs) of phosphor-converted (pc)-white LEDs, and also can estimate the residual color life.


Population structure and accompanying biota of the snail Turbo (Callopoma) funiculosus (Gastropoda: Turbinidae), on Socorro Island, Revillagigedo Archipelago, Mexico.
The porcelain snail, Turbo funiculosus, is a potential fishery resource that almost has not been studied or used commercially. In March of 1992, we sampled T. funiculosus in Bahia Binners and Bahia Blanca, Socorro Island, Revillagigedo Archipelago, using 25 m2 quadrants. We found that total length varies between 3.0 and 85 mm, proportional weight between 3.0 and 228 g, and density between 6.21 and 9.87 ind/m2. The largest organisms (over 50 mm) contain 30.6% of soft parts. Porcelain snail populations remain unexploited on Revillagedo Archipelago and could be a potential resource under an adequate management strategy.


[Mental symptoms perceptions of healthcare needs, and health care seeking behaviors, among children and adolescents in Mexico City].
To estimate the prevalence of behavioral and emotional symptoms in children and adolescents; to identify parents' perception of their children's healthcare needs; and to estimate the frequency of seeking mental healthcare services. A household survey was carried out between July and November 1995 in Mexico City. The survey was representative of persons aged 18 to 65 years. Study subjects were 1685 children 4 to 16 years of age and living in the same household as their parents. Informant parents provided data on their children regarding the presence of 27 symptoms, their perceptions of healthcare needs of symptomatic children, and mental healthcare-seeking behaviors. Logistic regression was used for statistical analysis. Symptoms were reported for half of the population. Twenty-five percent of children was perceived by their parents as in need for healthcare, and only 13% sought healthcare. Internalization symptoms were more frequently perceived by parents and led to seeking healthcare. Study findings suggest a delay in seeking mental healthcare in children and should be of value for clinicians, and to establish primary level mental healthcare policies and programs.


Streptococcus pneumoniae biofilm formation and dispersion during colonization and disease.
Streptococcus pneumoniae (the pneumococcus) is a common colonizer of the human nasopharynx. Despite a low rate of invasive disease, the high prevalence of colonization results in millions of infections and over one million deaths per year, mostly in individuals under the age of 5 and the elderly. Colonizing pneumococci form well-organized biofilm communities in the nasopharyngeal environment, but the specific role of biofilms and their interaction with the host during colonization and disease is not yet clear. Pneumococci in biofilms are highly resistant to antimicrobial agents and this phenotype can be recapitulated when pneumococci are grown on respiratory epithelial cells under conditions found in the nasopharyngeal environment. Pneumococcal biofilms display lower levels of virulence in vivo and provide an optimal environment for increased genetic exchange both in vitro and in vivo, with increased natural transformation seen during co-colonization with multiple strains. Biofilms have also been detected on mucosal surfaces during pneumonia and middle ear infection, although the role of these biofilms in the disease process is debated. Recent studies have shown that changes in the nasopharyngeal environment caused by concomitant virus infection, changes in the microflora, inflammation, or other host assaults trigger active release of pneumococci from biofilms. These dispersed bacteria have distinct phenotypic properties and transcriptional profiles different from both biofilm and broth-grown, planktonic bacteria, resulting in a significantly increased virulence in vivo. In this review we discuss the properties of pneumococcal biofilms, the role of biofilm formation during pneumococcal colonization, including their propensity for increased ability to exchange genetic material, as well as mechanisms involved in transition from asymptomatic biofilm colonization to dissemination and disease of otherwise sterile sites. Greater understanding of pneumococcal biofilm formation and dispersion will elucidate novel avenues to interfere with the spread of antibiotic resistance and vaccine escape, as well as novel strategies to target the mechanisms involved in induction of pneumococcal disease.


[Systems theory of the pulsed Doppler technic for blood flow measurement. II].
Following the discussion of the signal generation of an ideal pw Doppler system in the time and frequency domain using the means of the one-dimensional system theory in part 1 in this part 2 the main system parameters as resolution in space, and unambiguity ranges for space and velocity are explained and mathematical formula are derived for this.


The Determination of Some Microbiological and Chemical Features in Herby Cheese.
The objective of this study is to measure the amounts of biogenic amines, microbial counts, values of pH, titratable acidity, dry matter, and salt (%) in herby cheese, a very popular staple in the Turkish diet, and to evaluate the concentration of biogenic amines in terms of public health risks. A high-performance liquid chromatography (HPLC) method was used for the determination of eight biogenic amines in 100 herby cheeses sold in the local markets of Van. The bacterial load of the herby cheeses ranged between 4.0 and 8.90 log CFU/g for viable total aerobic mesophilic bacteria (TAMB), <1 and 7.0 log CFU/g for lactic bacteria (LAB), <1 and 6.08 log CFU/g for coliform bacteria, <1 and 5.81 log CFU/g for Enterobacteriaceae, <1 and 2.60 log CFU/g for Staphylococcus aureus, and 3.70 and 8.05 log CFU/g for yeasts and molds. The results obtained suggested significant changes in the pH, titratable acidity, dry matter, and salt contents of the examined herby cheese samples. The detection levels of biogenic amines in the samples ranged from <0.025 to 33.36 mg/kg for tryptamine, from <0.038 to 404.57 mg/kg for β-phenylethylamine, from 0.03 to 426.35 mg/kg for putrescine, from <0.039 to 1438.22 mg/kg for cadaverine, from <0.033 to 469 mg/kg for histamine, from <0.309 to 725.21 mg/kg for tyramine, from <0.114 to 1.70 mg/kg for spermidine, and from <0.109 to 1.88 mg/kg for spermine. As a result, these cheeses are fit for consumption in terms of the amounts of biogenic amines they contain.


[Experiences with the implantation of the 2-loop-medaillon-lens (author's transl)].
The experience with 100 primary implantations of the 2-loop-Medaillon-lens is reported. In 4% complications occured that decreased the visual outcome. The avoidance of complications is discussed. In the author's opinion implanting artificial lenses is a suitable method of visual rehabilitation - connected with little risk - of patients who are not suitable for correction with glasses or contact lenses. The indications and contraindications are discussed. The main indication is a onesided cataract.


Enteric locus of action of prokinetics: ABT-229, motilin, and erythromycin.
We investigated the in vivo and in vitro locus of actions of prokinetics: motilin, erythromycin, and ABT-229. The test substances were infused close intra-arterially in short segments of the jejunum in the intact conscious state. Each prokinetic acted on a presynaptic neuron and utilized at least one nicotinic synapse to stimulate circular muscle contractions. The final neurotransmitter at the neuroeffector junction was ACh. Motilin and erythromycin, but not ABT-229, also released nitric oxide. Each prokinetic utilized somewhat different subtypes of muscarinic, serotonergic, tachykininergic, and histaminergic receptors, except for the M(3) receptor, which was common to all of them. In contrast, none of the prokinetics stimulated contractions in mucosa-free or mucosa-attached muscle strips, or rings, even though methacholine or electrical field stimulation induced phasic contractions in all of them. The prokinetics also did not release ACh in longitudinal muscle-myenteric plexus preparations. Each prokinetic, however, decreased the length of enzymatically dispersed single cells. In conclusion, each prokinetic may act on a different subset of presynaptic neurons that converge on the postsynaptic cholinergic and nonadrenergic noncholinergic motoneurons. The presynaptic neurons may be impaired in the muscle bath environment.


Non-alcoholic fatty liver disease among visitors to a hepatitis awareness programme.
Non-alcoholic fatty liver disease (NAFLD) is rapidly becoming the most common liver disease worldwide. The present study aimed to evaluate the prevalence of NAFLD among the visitors screened on World Hepatitis Day 2010 and 2011. Attendees of the hepatitis awareness programme (n = 928) were offered screening to rule out liver ailments. The participants were evaluated on history, body mass index (BMI) measurement, blood tests for hepatitis B surface antigen (HBsAg), anti-hepatitis C virus (anti-HCV), alanine aminotransferase (ALT) levels and abdominal ultrasound (US). On ultrasonography, fatty liver was present in 142 of 806 subjects who were found negative for viral serology and had no history of regular alcohol intake. This accounted for 15.3% of all the visitors. Comparing them with the rest of the serology-negative alcohol-negative subjects, there were more housewives (p = 0.005) with fatty liver. They were older with mean age 43.3 +/- 12.1 years compared to 33.1 +/- 16.1 years (p = 0.000) in patients without fatty liver and had a higher BMI 28.6 +/- 6.0 kg/m2 vs. 22.2 +/- 5.3 kg/m2 (p = 0.000). Regression analysis suggested BMI as the only significant independent risk factor (p = 0.000). However, 32 subjects with fatty liver had BMI < 25 kg/m2 and 6 of them had diabetes mellitus. Thirty-one subjects (21.8%) with fatty liver had raised ALT level (p = 0.000) and possible non-alcoholic steatohepatitis (NASH) which accounts for 3.34% (31/928) of the total visitors. This analysis reveals that our general population is at risk of having NAFLD and NASH and these should be looked into even in non-obese persons.


14 Years of Polish Experience in Non-Invasive Prenatal Blood Group Diagnosis.
Blood cell antigens may cause maternal alloimmunization leading to fetal/newborn disorders. Non-invasive prenatal diagnostics (NIPD) of the blood group permits the determination of feto-maternal incompatibility. To evaluate 14 years of blood group NIPD at the Institute of Hematology and Transfusion Medicine (IHTM) in Warsaw. Plasma DNA from 536 RhD-negative, 24 Rhc-negative, 26 RhE-negative, 43 K-negative, and 42 HPA-1a-negative pregnant women was examined by real-time PCR to detect RHD, RHCE*c, RHCE*E, RHCE*C, KEL*01 and HPA*1A, respectively. We tested for CCR5, SRY or bi-allelic polymorphisms and quantified the total or fetal DNA. The results of fetal antigen status prediction by NIPD in all but one case (false-positive result of KEL*01) were correct taking neonate serology as a reference. It was confirmed that all negative results of NIPD contained fetal DNA except for four cases where there was no difference between the parents' polymorphisms. A fetal genotype compatible with the mother was determined in 25% of all pregnancies tested at the IHTM for the fetal blood group. These cases were not at risk of disease, so it was possible to avoid invasive procedures.


Output characteristics of DeVilbiss No. 40 hand-held jet nebulizers.
DeVilbiss No. 40 hand-held nebulizers are widely used for quantifying airway responsiveness in large populations using pharmacological agents. We examined the aerosol characteristics of five nebulizers. Within each device, the aerosol output and droplet size were reasonably stable over a wide range of bulb pressures, although there were considerable differences in output characteristics between nebulizers. The droplet size was very large compared to conventional aerosol delivery systems, with a mass median diameter greater than 10 microns for three of the five devices. Between 28-50% of the output was in particles sufficiently small for airway deposition (less than 6.2 microns). A more vigorous compression of the bulb caused a small increase output and a reduction in droplet size, resulting in a much bigger variation in the output of the respirable aerosol (less than 6.2 microns) with changes in bulb pressure. The loss due to evaporation was about 3.5%, causing a similar rise in the osmolality of the nebulizer solution. In view of the variable nebulizer output and the marked between-operator variation in bulb pressure, the characteristics of individual DeVilbiss No. 40 nebulizers should be evaluated by individual operators before use in clinical practice or research.


In vivo pulsed magneto-motive ultrasound imaging using high-performance magnetoactive contrast nanoagents.
Previously, pulsed magneto-motive ultrasound (pMMUS) imaging has been introduced as a contrast-agent-assisted ultrasound-based imaging modality capable of visualizing biological events at the cellular and molecular level. In pMMUS imaging, a high intensity pulsed magnetic field is used to excite cells or tissue labeled with magnetic nanoparticles. Then, ultrasound (US) imaging is used to monitor the mechanical response of the tissue to an externally applied magnetic field (i.e., tissue displacement). Signal to noise ratio (SNR) in pMMUS imaging can be improved by using superparamagnetic nanoparticles with larger saturation magnetization. Metal-doped magnetic nanoparticles with enhanced tunable nanomagnetism are suitable candidates to improve the SNR and, therefore, sensitivity of pMMUS imaging, which is essential for in vivo pMMUS imaging. In this study, we demonstrate the capability of pMMUS imaging to identify the presence and distribution of zinc-doped iron oxide nanoparticles in live nude mice bearing A431 (human epithelial carcinoma) xenograft tumors.


Stable brain ATM message and residual kinase-active ATM protein in ataxia-telangiectasia.
The gene that is mutated in ataxia-telangiectasia (A-T), ATM, is catalytically activated in response to DNA damage. Yet a full accounting for the CNS deficits in human A-T or its mouse models remains elusive. We have analyzed the CNS phenotypes of two mouse Atm alleles--Atm(tm1Bal) (Bal) and Atm(tm1Awb) (Awb). Neither mutant has detectable mRNA or protein in peripheral tissues. In brain, although Bal/Bal mice have no ATM protein, they have nearly normal amounts of Atm mRNA. Bal/Bal neurons exhibit extensive cell cycle reentry and degeneration in both cortex and cerebellum. Unexpectedly, in Awb/Awb mice a novel mRNA is found in which the engineered mutation is excised. This mRNA is apparently translated and produces a catalytically active ATM protein that responds to DNA damage by phosphorylating p53 and Chk2. Prompted by these results, we examined eight cases of human A-T and found evidence for residual ATM protein in seven of them. These findings offer important new insights into the human disease and the role of brain ATM activity in the severity of the neurological symptoms of A-T.


Palladium-catalyzed cascade reaction of alpha,beta-unsaturated sulfones with aryl iodides.
Unlike traditionally used acyclic 1,2-disubstituted alkenes, the reaction of alpha,beta-unsaturated phenyl sulfones with aryl iodides under Heck reaction conditions (Pd(OAc)(2) as catalyst, Ag(2)CO(3) as base in DMF at 120 (0)C) takes place mainly by a cascade process, involving one unit of the alkene and three units of the aryl iodide, to afford a substituted 9-phenylsulfonyl-9,10-dihydrophenanthrene. The dominant formation of this 3:1 coupling product, instead of the Heck trisubstituted olefin, shows that aromatic C-H bond activation processes can compete with the usually fast syn beta-hydrogen elimination step in the Heck arylation of an acyclic olefin. The structural scope of this palladium-catalyzed cascade arylation of alpha,beta-unsaturated sulfones has proved to be wide with regard to substitution at the beta-position (alkyl, aryl, or alkenyl substitution), substitution at the sulfone unit (alkyl or phenyl sulfones), and configuration at the CdoublebondC bond (trans or cis). Moreover, although less favored than in the case of the arylation of alpha,beta-unsaturated sulfones, similarly substituted 9,10-dihydrophenanthrenes have also been obtained in the case of alpha,beta-unsaturated phosphine oxides and alpha,beta-unsaturated phosphonate esters. A Pd(0)-Pd(II)-Pd(IV) mechanistic pathway involving the successive formation of highly electrophilic sigma-alkylpalladium intermediates and palladacycles is proposed for this multicomponent arylation.


Learning Disabilities and Medical Students.
This article was migrated. The article was marked as recommended. Background: Learning disabilities (LD) are a mixed group of disorders exhibited by substantial difficulties in the achievement and use of listening, speaking, reading, writing, reasoning, or mathematical skills. Objective: The objective of this paper is to highlight different learning disabilities and their effects on medical students and suggest the best assessment strategy for such students. Methods: The medical education literature was searched for articles related to learning disabilities and how they effect medical students. Results and Conclusion: Learning disability constitutes to be challenging for the student as well as the faculty because apparently, there is no disability to be seen. It is difficult to diagnose and sometimes it remains unidentified till late adulthood when the compensatory mechanisms crash down leaving the student in despair and low self-confidence. The identification of such a disorder requires appropriate personnel to diagnose such condition which are currently not available in the medical schools. Literature shows that multiple choice questions (MCQs) are the best method of choice in assessment of students with learning disability as it does not discriminate between students with and without LD.