Melatonin pretreatment improves gastric mucosal blood flow and maintains intestinal barrier function during hemorrhagic shock in dogs.
Melatonin improves hepatic perfusion after hemorrhagic shock and may reduce stress-induced gastric lesions. This study was designed to investigate whether pretreatment with melatonin may influence gastric mucosal microcirculatory perfusion (μflow), oxygenation (μHbO2 ), or intestinal barrier function during physiological and hemorrhagic conditions in dogs. In a randomized crossover study, five anesthetized foxhounds received melatonin 100 μg kg-1 or vehicle (ethanol 5%) intravenously in the absence or presence of hemorrhagic shock (60 minutes, -20% blood volume). Systemic hemodynamic variables, gastric mucosal perfusion, and oxygenation were recorded continuously; intestinal barrier function was assessed intermittently via xylose absorption. During hemorrhagic shock, melatonin significantly attenuated the decrease in μflow, compared with vehicle (-19±9 vs -43±10 aU, P<.05), without influence on μHbO2 . A significant increase in xylose absorption was detected during hemorrhage in vehicle-treated dogs, compared with sham-operated animals (13±2 vs 8±1 relative amounts, P<.05); this was absent in melatonin-treated animals (6±1 relative amounts). Melatonin did not influence macrocirculation. Melatonin improves regional blood flow suggesting improved oxygen delivery in gastric mucosa during hemorrhagic shock. This could provide a mechanism for the observed protection of intestinal barrier function in dogs.


Associations between sexually submissive and dominant behaviors and sexual function in men and women.
The current study investigated the associations between sexually submissive and dominant behaviors and sexual dysfunction in Finnish men and women. We analyzed three population-based data sets from 2006, 2009, and 2021-2022, including 29,821 participants in total. Participants filled out a questionnaire about their sexually submissive and dominant behaviors, Sexual Distress Scale, Checklist for Early Ejaculation Symptoms and International Index of Erectile Function Questionnaire-5 (men), and Female Sexual Function Index (women). Pearson Correlations showed that for both sexes, sexually submissive (men: r = 0.119, p < 0.001; women: r = 0.175, p < 0.001) and dominant (men: r = 0.150, p < 0.001; women: r = 0.147, p < 0.001) behaviors were both associated with more sexual distress. However, for men, sexually submissive (r = -0.126, p < 0.001) and dominant behaviors (r = -0.156, p < 0.001) were associated with less early ejaculation symptoms. Both sexually submissive (r = 0.040, p = 0.026) and dominant behaviors (r = 0.062, p < 0.001) were also associated with better erectile function while sexually dominant behavior alone was associated with better orgasmic function (r = 0.049, p = 0.007), intercourse satisfaction (r = 0.068, p < 0.001), and overall satisfaction (r = 0.042, p = 0.018). For women, both sexually submissive (r = 0.184, p < 0.001) and dominant behaviors (r = 0.173, p < 0.001) were also associated with better overall female sexual function. One possible explanation is that these individuals have a clear idea of what they prefer sexually facilitating arousal. Particularly, sexually submissive behavior may reduce high-level self-awareness and, in this way, contribute to reduced performance anxiety. However, non-normative interests seem to simultaneously result in increased sexual distress probably due to the absence of self-acceptance. Further research about the causal mechanisms between non-normative sexual interest and sexual function is needed.


Membrane remodulation and hyperactivation are impaired in frozen-thawed sperm of low-fertility bulls.
Bulls used in artificial insemination programmes worldwide undergo quality control checks, which are typically based on the evaluation of sperm motility and morphology. Despite this, some bulls can have lower than expected field fertility and the reasons for this remain to be elucidated. Here we hypothesised that sperm from bulls of varying fertility will differ in their ability to undergo capacitation-related events including an increase in membrane fluidity, protein tyrosine phosphorylation, hyperactivation and the acrosome reaction. Firstly, we used frozen-thawed semen from 10 high-fertility (HF) and 10 low-fertility (LF) bulls, and subjected them to in vitro capacitating conditions, following which sperm viability, membrane fluidity, acrosome integrity and protein tyrosine phosphorylation were assessed using flow cytometry. We then assessed the ability of sperm to undergo hyperactivation (induced using caffeine) utilising computer-assisted sperm analysis, and the acrosome reaction (induced using calcium ionophore) using flow cytometry. When sperm were incubated in capacitating conditions, a higher percentage of viable sperm from HF bulls exhibited high membrane fluidity when compared to LF bulls (8.8 ± 0.8% and 5.8 ± 1.2%, respectively; mean ± standard error; P < 0.05). There was no difference between fertility groups in the percentage of acrosome-reacted sperm following the incubation in in vitro capacitating conditions or following the induction of the acrosome reaction using calcium ionophore. However, more sperm from HF bulls became hyperactive in response to caffeine stimulation than sperm from LF bulls (21.6 ± 2.5% versus 14.1 ± 2.4%, respectively; mean ± standard error; P < 0.05). Taken together, sperm from LF bulls had an impaired ability to undergo membrane remodulation and to hyperactivate when induced in vitro. These are key events in the journey of sperm along the female reproductive tract and in the interaction with the oocyte and thus could explain the lower field fertility exhibited by some bulls.


Gain-of-function mutant p53 together with ERG proto-oncogene drive prostate cancer by beta-catenin activation and pyrimidine synthesis.
Whether TMPRSS2-ERG fusion and TP53 gene alteration coordinately promote prostate cancer (PCa) remains unclear. Here we demonstrate that TMPRSS2-ERG fusion and TP53 mutation / deletion co-occur in PCa patient specimens and this co-occurrence accelerates prostatic oncogenesis. p53 gain-of-function (GOF) mutants are now shown to bind to a unique DNA sequence in the CTNNB1 gene promoter and transactivate its expression. ERG and β-Catenin co-occupy sites at pyrimidine synthesis gene (PSG) loci and promote PSG expression, pyrimidine synthesis and PCa growth. β-Catenin inhibition by small molecule inhibitors or oligonucleotide-based PROTAC suppresses TMPRSS2-ERG- and p53 mutant-positive PCa cell growth in vitro and in mice. Our study identifies a gene transactivation function of GOF mutant p53 and reveals β-Catenin as a transcriptional target gene of p53 GOF mutants and a driver and therapeutic target of TMPRSS2-ERG- and p53 GOF mutant-positive PCa.


Quantification of tricuspid regurgitation area by 3-dimensional color Doppler echocardiography considering different clinical settings.
The precise assessment of tricuspid regurgitation (TR) using 2D imaging techniques may be associated with significant difficulties due to the nonround regurgitation area. Direct analysis of the regurgitation area by 3D color Doppler echocardiography at the vena contracta (3D VCA) has the potential to adequately quantify even complex TR. This study compared 3D VCA for quantification of the TR with the regurgitant area determined by proximal isovolumetric convergence method (PISA-EROA) considering different clinical settings. In 95 patients with TR of different severity, the regurgitant orifice area was determined by 3D color Doppler echocardiography and by PISA-EROA. Using 3D color Doppler echocardiography, the regurgitant orifice area was determined three times in each patient considering 3 datasets. Mean 3D VCA was 0.27 ± 0.14, 0.27 ± 0.13, and 0.29 ± 0.14 cm2 , respectively, as determined by three separate measurements in each of the 95 patients. There was a mean relative deviation between the three measurements in each patient of 12.4 ± 14.9%. The regurgitant orifice area using the PISA method was 0.28 ± 0.14 cm2 . There was a mean difference of 0.07 cm2 (95% CI -0.124 to 0.138 cm2 ) between 3D VCA and PISA-EROA. The correlation between 3D VCA and PISA-EROA was r = .88 (P < .001). Considering a grading of TR severity in grade I (regurgitant area < 0.2 cm2 ), grade II (area 0.2-0.4 cm2 ), and grade III (area > 0.4 cm2 ), there was a good agreement between severity grade determined by 3D VCA and severity grade determined by PISA-EROA (kappa 0.71). The analysis of the VCA of a TR using 3D color Doppler echocardiography is an alternative method to determine the regurgitant severity with good agreement to the PISA method.


Effects of melanocortin receptor ligands on thyrotropin-releasing hormone release: evidence for the differential roles of melanocortin 3 and 4 receptors.
The hypothalamic melanocortin system is important in the central regulation of food intake and body weight. We have previously demonstrated that intracerebroventricular administration of alpha-melanocyte stimulating hormone (alpha-MSH), a nonselective MC3 and MC4 receptor agonist, stimulated plasma thyroid-stimulating hormone, and agouti-related protein (AgRP), an MC3 and MC4 receptor antagonist, suppressed it. In this study, we investigated the effects of MC3 and MC4 receptor (MC3-R and MC4-R) selective agonists and antagonists on the release of thyrotropin-releasing hormone (TRH) from hypothalamic explants in vitro. alpha-MSH stimulated TRH release from the rat hypothalamic explants (alpha-MSH 100 nm 230 +/- 22.9% basal, P < 0.005). In contrast, gamma 2-MSH, a selective MC3-R agonist, suppressed TRH release (gamma 2-MSH 10 microns 76.2 +/- 7.4% basal, P < 0.05). AgRP (83-132), a nonselective MC3/4-R antagonist, induced no change in TRH release whilst JKC-363 (cyclic [Mpr11, D-Nal14, Cys18, Asp22-NH2]-beta-MSH 11-22), a selective MC4-R antagonist, suppressed it (JKC-363 10 nm 57.2 +/- 11.5% basal, P < 0.05). Both AgRP (83-132) and JKC-363 blocked alpha-MSH stimulated TRH release but only AgRP (83-132) blocked the inhibitory effect of gamma 2-MSH on TRH release. These data suggest differential roles for the MC3 and MC4 receptors in TRH release; MC3-R agonism inhibiting and MC4-R agonism stimulating TRH release.


Association study between the D10S1423 microsatellite marker and Alzheimer's disease.
Several studies have reported conflicting results concerning the genetic association between Alzheimer's disease (AD) and the microsatellite marker D10S1423 on chromosome 10p12-14. In an ethnically homogeneous German population of 422 patients with AD and 254 cognitively healthy controls, the 238-bp allele of the D10S1423 marker showed a weak, but after correction for multiple testing no longer significant association with AD (p = 0.015, uncorrected; p = 0.11, corrected). These findings do not support the presence of a relevant susceptibility locus for AD on chromosome 10p12-14.


Wiberg Type III patellae and J-sign during extension compromise outcomes of total knee arthroplasty without patellar resurfacing.
Understanding the risk factors associated with postoperative pain and worse outcome can guide surgeons on whether primary patellar resurfacing is warranted during total knee arthroplasty (TKA). The purpose of this study was to determine whether clinical scores and pain after TKA without patellar resurfacing are correlated with patellar shape and postoperative patellar position and kinematics. Radiographs as well as anterior knee pain according to the Visual Analogue Scale (pVAS) were collected pre- and postoperatively for 100 knees aged 68 ± 7.7 years that received uncemented TKA without patellar resurfacing. At a minimum follow-up of 12 months the Forgotten Joint Score (FJS), the Oxford Knee Score (OKS) as well as the flexion range of motion and the presence of J-sign during active extension were recorded. Uni- and multivariable linear regression analyses were performed to determine associations between the collected clinical scores and patient demographic and radiographic data. Postoperative OKS (79 ± 14.4) was worse for Wiberg Type III patellae (β = -11.4, P = .020, compared with Type II). Anterior pVAS (2 ± 2) was greater in knees with J-sign during extension (β = 2.8, P < .001). None of the other radiographic measurements (patellar tilt, congruence angle and lateral patellar displacement) were correlated with postoperative OKS or anterior pVAS. Incongruent patellar shape (Wiberg Type III) is associated with worse clinical scores, and abnormal kinematics (J-sign) with increased pain after TKA without patellar resurfacing. The authors therefore recommend routine resurfacing for Wiberg Type III patellae, though further studies are required to confirm whether resurfacing truly improves clinical scores and pain in this subgroup.


Pacemaker therapy for academic medicine.
Both the clinical practice and the administrative environments of academic medicine have become progressively more complex. Although management methods for dealing with the administrative intricacies are commonly not part of academic training, guidelines may be derived from medical practice. The authors identify specific parallels and analogies between recent developments in the design of pacemakers and the requirements of organizations that implant them.


Intracranial paragangliomas versus schwannomas: Role of dynamic susceptibility contrast perfusion and diffusion MRI.
Differentiating paragangliomas from schwannomas and distinguishing sporadic from neurofibromatosis type 2 (NF 2)-related schwannomas is challenging but clinically important. This study aimed to assess the utility of dynamic susceptibility contrast perfusion MRI (DSC-MRI) and diffusion-weighted imaging (DWI) in discriminating infratentorial extra-axial schwannomas from paragangliomas and NF2-related schwannomas. This retrospective study included 41 patients diagnosed with paragangliomas, sporadic schwannomas, and NF2-related schwannomas in the infratentorial extra-axial space between April 2013 and August 2021. All cases had pretreatment DSC-MRI and DWI. Normalized mean apparent diffusion coefficient (nADCmean), normalized relative cerebral blood volume (nrCBV), and normalized relative cerebral blood flow (nrCBF) were compared between paragangliomas and schwannomas and between sporadic and NF2-related schwannomas as appropriate. nrCBV and nrCBF were significantly higher in paragangliomas than in sporadic/NF2-related schwannomas (nrCBV: median 11.5 vs. 1.14/3.74; p < .001 and .004, nrCBF: median 7.43 vs. 1.13/2.85; p < .001 and .007, respectively), while nADCmean were not. The corresponding diagnostic performances were area under the curves (AUCs) of .99/.92 and 1.0/.90 with cutoffs of 2.56/4.22 and 1.94/3.36, respectively. nADCmean were lower, and nrCBV and nrCBF were higher in NF2-related than in sporadic schwannomas (nADCmean: median 1.23 vs. 1.58, nrCBV: median 3.74 vs. 1.14, nrCBF: median 2.85 vs. 1.13; all p < .001), and the corresponding diagnostic performances were AUCs of .93, .91, and .95 with cutoffs of 1.37, 2.63, and 2.48, respectively. DSC-MRI and DWI both can aid in differentiating paragangliomas from schwannomas and sporadic from NF2-related schwannomas.


The human antibody repertoire to infectious agents: implications for disease pathogenesis.
Antibodies are critical entities used for the protection of an individual organism against infection and disease. Through a complex series of events genes rearrange to encode repertoires of molecules, which are tested for their ability to identify foreign molecules. In this report, we discuss factors defined by the individual as well as by infectious organisms that shape the antibody repertoire. A more thorough understanding of the interplay between these factors will eventually allow us to elucidate the underlying mechanisms of disease susceptibility following infection, and to design potent vaccines and other immunomodulating reagents.


Golden Jubilee of Singapore National Institute of Chemistry (1970-2020): Celebrating its Partnership with Wiley-VCH.
This year Singapore National Institute of Chemistry (SNIC) is celebrating its golden jubilee (1970-2020). Wiley-VCH has been a steadfast partner accompanying the rapid rise of chemistry research in Singapore. In celebration of this golden jubilee, we highlight 50 significant papers published in Angewandte Chemie by scholars currently based in Singapore, covering the widest possible spectrum of chemistry research.


"Nocturia and obstructive sleep apnea syndrome: A systematic review".
Lower urinary tract symptoms represent a significant public health problem worldwide, impairing patients' quality of life, especially in elderly people. Among LUTS, nocturia is assessed as the most experienced entity related to several disorders such as sleep disorders and/or obstructive sleep apnea syndrome (OSAS). Among OSAS patients, nocturia stands as a bothersome symptom that increases alongside with the OSAS severity. However, despite the nocturia and OSAS shared a long-acknowledged link, the causes, and the pathophysiology for development of nocturia in OSAS have remained largely unexamined. Generally, the patients with OSAS experienced nocturia due to easy waking or increased bladder filling. However, nor the effect of treatment on management of nocturia in OSAS patients are well-established.


High concentrations of STOP protein induce a microtubule super-stable state.
We have previously shown that mammalian brain crude extracts contained two classes of stable microtubules: "cold stable" and "super-stable" microtubules. We now find that both species are generated by a single protein factor (STOP protein) in a dose dependent manner. These results show that STOP protein action can be extreme, inducing resistance to -80 degrees C or to sonication and that no other factor seems to be required to account for the various subclasses of highly stable microtubules in brain. Finally, the rapid procedure described for the preparation of purified "super-stable microtubules" should be useful for the obtention of fractions with high STOP protein activity.


3D-QSAR study of Chk1 kinase inhibitors based on docking.
Checkpoint kinase 1 (Chk1), a kind of a serine/threonine protein kinase, plays a significant role in DNA damage-induced checkpoints. Chk1 inhibitors have been demonstrated to abrogate the S and G2 checkpoints and disrupt the DNA repair process, which results in immature mitotic progression, mitotic catastrophe, and cell death. Normal cells remain at the G1 phase via p53 to repair their DNA damages, and are less influenced by the abrogation of S and G2 checkpoint. Therefore, selective inhibitors of Chk1 may be of great therapeutic value in cancer treatment. In this paper, in order to understand the structure-activity relationship of macro-cyclic urea Chk1 inhibitors, a study combined molecular docking and 3D-QSAR modeling was carried out, which resulted in two substructure-based 3D-QSAR models, including the CoMFA model (r(2), 0.873; q(2), 0.572) and CoMSIA model (r(2), 0.897; q(2), 0.599). The detailed microscopic structures of Chk1 binding with inhibitors were performed by molecular docking. Two docking based 3D-QSAR models were developed (CoMFA with r(2), 0.887; q(2), 0.501; CoMSIA with r(2), 0.872; q(2), 0.520). The contour maps obtained from the 3D-QSAR models in combination with the docked binding structures would be helpful to better understand the structure-activity relationship. All the conclusions drawn from both the 3D-QSAR contour maps and molecular docking were in accordance with the experimental activity dates. The results suggested that the developed models and the obtained CHk1 inhibitor binding structures might be reliable to predict the activity of new inhibitors and reasonable for the future drug design.


Ergonomics in IR.
Ergonomic research in the field of interventional radiology remains limited. Existing literature suggests that operators are at increased risk for work-related musculoskeletal disorders related to the use of lead garments and incomplete knowledge of ergonomic principles. Data from existing surgical literature suggest that musculoskeletal disorders may contribute to physician burnout and female operators are at a higher risk of developing musculoskeletal disorders. This review article aims to summarize the existing ergonomic challenges faced by interventional radiologists, reiterate existing solutions to these challenges, and highlight the need for further ergonomic research in multiple areas, including burnout and gender.


Busulfan produces efficient human cell engraftment in NOD/LtSz-Scid IL2Rgamma(null) mice.
Xenografting immunodeficient mice after low-dose irradiation has been used as a surrogate human hematopoietic stem cell (HSC) assay; however, irradiation requires strict and meticulous animal support and can produce significant mortality rates, limiting the usefulness of this model. In this work, we examined the use of parenteral busulfan as an alternative conditioning agent. Busulfan led to dose-dependent human HSC engraftment in NOD/LtSz-scid/IL2Rgamma(null) mice, with marked improvement in survival rates. Terminally differentiated B and T lymphocytes made up most of the human CD45+ cells observed during the initial 5 weeks post-transplant when unselected cord blood (CB) products were infused, suggesting derivation from existing mature elements rather than HSCs. Beyond 5 weeks, CD34+-enriched products produced and sustained superior engraftment rates compared with unselected grafts (CB CD34+, 65.8% +/- 5.35%, vs. whole CB, 4.27% +/- 0.67%, at 24 weeks). CB CD34+ group achieved significantly higher levels of engraftment than mobilized CD34+-enriched peripheral blood (PB CD34+). At 8 weeks, all leukocyte subsets were detected, yet human red blood cells (RBCs) were not observed. Transfused human red cells persisted in the chimeric mice for up to 3 days; an accompanying rise in total bilirubin suggested hemolysis as a contributing factor to their clearance. Recipient mouse-derived human HSCs had the capacity to form erythroid colonies in vitro at various time points post-transplant in the presence of human transferrin (Tf). When human Tf was administered singly or in combination with anti-CD122 antibody and human cytokines, up to 0.1% human RBCs were detectable in the peripheral blood. This long evasive model should prove valuable for the study of human erythroid cells.


Effect of a leaf-shredding invertebrate on organic matter dynamics and phosphorus spiralling in heterotrophic laboratory streams.
The effect of invertebrate shredders on organic matter dynamics and phosphorus spiralling was studied over a 30-week period in laboratory streams. The streams were fed by groundwater, layered with cobble and gravel from a natural stream, covered with opaque material to eliminate algal growth, and initially contained 195 g/m2 of autumn-shed leaves. Four weeks after leaf addition, leaf-shredding snails (Goniobasis clavaeformis) were added to each of three streams in densities of 75, 220, and 800/m2. A fourth stream contained no snails and served as a control.Presence of snails increased the loss rates of coarse particulate organic matter (CPOM) and total organic matter (TOM), primarily by increasing leaf fragmentation and seston export. Although snail feeding increased specific metabolism of microbes associated with CPOM and cobble surfaces, it was not enough to compensate for reduction in bacterial cell numbers per unit surface area and in stream TOM. Consequently mineralization of detritus and whole stream phosphorus utilization rate were maximum in the stream with no snails and decreased with increasing snail density. From previous simulations of a stream model based on the nutrient spiralling concept, we predicted that there should be an intermediate shredder density which would minimize phosphorus spiralling length (maximize phosphorus utilization) in a natural stream nearby. Our current results conflict with the model-based predictions primarily because the increase in microbial metabolism was less important than reduction in bacterial cell numbers and total benthic organic matter resulting from snail feeding. Although our results indicate macroinvertebrate shredders reduce phosphorus utilization in headwater streams, shreders may increase nutrient utilization downstream where riparian inputs are lower, thus linking low- and high-order streams.


Age-related changes in the fibre composition of equine peripheral nerve.
Transverse sections of the lateral palmar nerve from a group of horses free from neuromuscular disease were assessed quantitatively, with emphasis on differences in the fibre population related to age. Morphometric analysis of the population of myelinated and unmyelinated fibres was performed. Changes were identified, there being a loss of larger myelinated fibres in older horses. The relationship of myelin thickness to axon diameter and the unmyelinated fibre population did not alter with age. The lateral palmar nerve is suitable for biopsy and electrophysiological study in the horse, but age related changes must be appreciated in the interpretation of results.


Local versus general anaesthesia in carotid surgery. A prospective, randomised study.
A randomised, prospective study was performed to compare local (LA) and general anaesthesia (GA) in carotid surgery with special emphasis on complications and the need for intra-operative shunting. Fifty-six patients were randomised to LA and 55 to GA. Eight patients in the LA group required a GA for various reasons. During the same period 14 patients were not randomised. Seven perioperative neurological deficits occurred (5.6%), four in the LA group, two in the GA group, and one in the non-randomised group (NS). Selective shunting was used, in the Ga group according to stump pressure or in cases with a previous stroke and in the LA group according to the appearance of neurological symptoms. In the GA group 25 patients were shunted and in the LA group five patients (P less than 0.001) needed a shunt. If strict pressure criteria for shunting had been used in the LA patients, ten would have been shunted and three of the patients who developed symptoms during clamping would not have been shunted. During surgery the highest recorded systolic pressure was significantly higher in the LA group (210 mmHg versus 173 mmHg, P less than 0.001). LA for carotid endarterectomy is comparable with general anaesthesia regarding peroperative complications but produces significantly higher blood pressures than general anaesthesia. On the other hand it allows the possibility of neurologic monitoring of the patient and leads to significantly less use of an intra-operative shunt.


Generalized loss of inhibitory innervation reverses serotonergic inhibition into excitation in a rabbit model of TNBS-colitis.
1. Inflammation may affect subpopulations of neurons of the myenteric plexus. 2. In the present study the effect of trinitrobenzene sulphonic acid (TNBS) induced colitis on nitrergic, purinergic and adrenergic inhibitory neurotransmission was studied as well as the consequences of the related changes on the response of 5-HT agonists using these neurotransmitters to mediate their effect. 3. Strips from normal and colitis rabbits (135 mg kg(-1) TNBS) were subjected to electrical field stimulation (EFS, 0.3 ms, 6V, 0.5 - 32 Hz, 10 s train). The response was measured isometrically in the absence or presence of L-NAME, suramin, guanethidine, the 5-HT agonists (5-HT(1/5A/7): 5-carboxamidotryptamine (5-CT), 5-HT(2): alpha-methyl-5-HT, 5-HT(3): 2-methyl-5-HT, 5-HT(4): 5-methoxytryptamine (5-MeOT)) or a combination. 4. In normal strips L-NAME (1 - 32 Hz), suramin (0.5 - 2, 8 Hz) and guanethidine (4, 16, 32 Hz) increased the response to EFS. This effect was abolished in inflamed strips and was accompanied by a decrease in nNOS expression. 5. In normal strips all 5-HT agonists induced pronounced (5-CT, alpha-methyl-5-HT) or small (2-methyl-5-HT, 5-MeOT) inhibitory neural responses. In inflamed strips this was reversed to cholinergic excitatory responses. 6. The effect of inflammation on the 5-HT(4) response was mimicked by preincubation of normal strips with L-NAME or suramin. Accordingly, in inflamed strips L-NAME or suramin did not affect the excitatory effects of 5-MeOT. 7. TNBS-colitis abolishes nitrergic, purinergic and adrenergic neurotransmission. This reverses serotonergic inhibition into excitation.


Receptor specificity of Ia-restricted T lymphoblasts activated against trinitrobenzene sulfonate-coupled spleen cells: recognition of distinct trinitrophenyl and Ia moieties.
The receptor specificity of H-2-restricted T lymphoblasts activated against trinitrobenzene sulfonate (TNBS)-coupled spleen cells was examined using an antigen binding assay. A population of Lyt-1+,2-T lymphoblasts acquired syngeneic Ia determinants during 4 days of primary culture with hapten-coupled stimulator cells. Syngeneic Ia was not reexpressed after trypsin treatment of the T cells, but was found after incubation with soluble Ia shed from lipopolysaccharide-activated blasts. Self-Ia binding was specific in that Lyt-1+,2- but not Lyt-1-,2+ cells acquired the antigen, and in that self-Ia bound more effectively than allogeneic Ia material. To determine the relationship of self-Ia binding to the recognition of foreign antigen, the binding of trinitrophenyl (TNP)-coupled plasma membrane vesicles by TNP-specific T cells was studied. TNP-vesicle binding occurred via TNP and H-2(Ia) molecules on the vesicles in that binding was inhibited with antibodies against TNP or H-2(Ia) molecules but not non-major histocompatibility complex (e.g., Ly-6.2) molecules on the vesicles. Complete inhibition of TNP-vesicle binding by an Iak-restricted TNP-specific T-cell line occurred with soluble TNP-lysine, but not an unrelated hapten, N-iodoacetyl-N-(5-sulfonic-1-naphthyl)ethylenediamine (I-AED)-cysteine. Conversely, I-AED-cysteine, but not TNP-lysine, inhibited binding of I-AED-coupled B6 vesicles by B6 anti-I-AED T cells. Significant, but weak inhibition of TNP-vesicle binding by the anti-TNP line was observed with glycoprotein preparations containing partially purified self-Ia molecules. However, inhibition was specific for I-Ak molecules, in that inhibition was lost after removal of I-Ak molecules from the glycoprotein preparation, and very little inhibition occurred with soluble glycoproteins prepared from thymocytes which contained very little Ia material or from LPS blasts of an unrelated H-2 haplotype. These results suggest a recognition model in which TNP and Ia determinants are recognized by neighboring receptor combining sites.


Season, temperature and blood pressure: a complex interaction.
An increase in blood pressure values measured during winter either in the office, at home, or at ambulatory blood pressure monitoring was consistently observed. Besides potentially contributing to increase the risk for cardiovascular events during the cold season, long term blood pressure variations can influence results of clinical trials, epidemiological surveys, and require personalized management of antihypertensive medications in the single patient. Those variations are often considered as an effect of climate, due to the close correlation observed in various countries and in different settings between temperature and blood pressure among children, adults, and specially the elderly. However, obtaining true measurements of exposition is a main problem when investigating the effects of climate on human health especially when the aim is to disentangle the effects of climate from those of seasonality. The aim of the present note is not to provide a complete review of the literature demonstrating the implications of seasonal blood pressure changes in the clinical and experimental setting; rather it is to consider methodological aspects useful to investigate the interaction between seasonality and temperature on blood pressure and to make health care providers aware of the implications of environmental factors on blood pressure in clinical and research settings.


Induction of premature senescence in human lung cancer by fungal immunomodulatory protein from Ganoderma tsugae.
Purified recombinant fungal immunomodulatory protein from Ganoderma tsugae (reFIP-gts) has anti-telomerase effects in human lung adenocarcinoma A549 cells. However, how reFIP-gts affects cancer cell fates remains unclear. Here, we demonstrated that reFIP-gts-treated lung cancer cells are arrested at G1 phase by flow cytometry and possess morphological phenotype consistent with cellular senescence. The senescent nature of these cells was supported by positive staining for senescence-associated beta-galactosidase activity and increased lysosomal content in A549 and CaLu-1 lung cancer cells. Arrest of cells at G1 appears to be the key means through which reFIP-gts induces premature cellular senescence in A549 cells. Finally, reFIP-gts- treated A549 cells grew more slowly and formed significantly fewer cell colonies in soft agar than untreated A549 cells. In an in vivo mouse model, A549 cells treated with reFIP-gts grew significantly slower than cells treated with PBS alone, confirming that lung tumor can be inhibited by reFIP-gts. The use of reFIP-gts may be a powerful new strategy for chemoprevention and antineoplastic therapy.


PI3K signalling: the path to discovery and understanding.
Over the past two decades, our understanding of phospoinositide 3-kinases (PI3Ks) has progressed from the identification of an enzymatic activity associated with growth factors, GPCRs and certain oncogene products to a disease target in cancer and inflammation, with PI3K inhibitors currently in clinical trials. Elucidation of PI3K-dependent networks led to the discovery of the phosphoinositide-binding PH, PX and FYVE domains as conduits of intracellular lipid signalling, the determination of the molecular function of the tumour suppressor PTEN and the identification of AKT and mTOR protein kinases as key regulators of cell growth. Here we look back at the main discoveries that shaped the PI3K field.


Ocular and periorbital anthropometric measurements in term Turkish newborns.
This study was conducted in an effort to determine the normal landmarks in the eyelids and eyebrows of newborns and the relationships between them. We also correlated all measurements with gender and head circumferences. The anthropometric measurements were performed on 86 normal term newborns (40 males and 46 females) in the first 48 hours after delivery. There was no significant difference between either sex, or between the right and left eyes. In contrast, a statistically significant positive correlation was found between the head circumference and the horizontal fissure length (r = 0.42, p < 0.01). In addition, a significant correlation was found between the upper eyelid crease distance and the horizontal and the vertical fissure lengths (r = 0.27, p < 0.01; r = 0.22, p < 0.05, respectively). The results indicate that the normal eyelid crease location may be used as a landmark for guidance to plan surgical and reconstructive procedures.


Silent Information Regulator 1 Negatively Regulates Atherosclerotic Angiogenesis via Mammalian Target of Rapamycin Complex 1 Signaling Pathway.
This study aimed to investigate the interactions between silent information regulator 1 (SIRT1) and mammalian target of rapamycin (mTOR) in intraplaque angiogenesis and their potential mechanisms through in vivo and in vitro studies. An atherosclerosis model was established in 12 rabbits on a high-cholesterol diet. The rabbits were equally divided into 3 groups: a control group (high-lipid diet), RAP group (high-lipid diet supplemented with rapamycin) and RAP + NAM group (high-lipid diet supplemented with rapamycin and nicotinamide). At the end of 4 weeks, the area of plaques in the aorta was determined and the protein expression of CD31 and vascular endothelial growth factor (VEGF) was detected through hematoxylin and eosin staining and immunohistochemical staining, respectively. For in vitro study, a hypoxia model was established in human umbilical vein endothelial cells (HUVECs) by using the chemical method (CoCl2). The MTT assay, scratch assay and tube formation assay were performed to evaluate the proliferation and angiogenesis abilities of HUVECs. Reverse transcription polymerase chain reaction was used to examine the mRNA levels of SIRT1, hypoxia-inducible factor-1α (HIF-1α), mTOR and p70 ribosomal S6 kinase (p70S6K). Western blotting was used to examine the protein levels of SIRT1, HIF-1α, mTOR, p-mTOR, p-raptor and p-p70S6K. The results of the in vivo study indicated a significant inhibitory effect of rapamycin on plaque size and intraplaque angiogenesis (0.05 ± 0.02mm2 versus 5.44 ± 0.50mm2, P < 0.05). This effect was attenuated by nicotinamide (0.76 ± 0.15mm2 versus 0.05 ± 0.02mm2, P < 0.05). Compared with the RAP group, CD31- and VEGF-positive vessels were abundant in the RAP + NAM group. The RAP group showed lower expression of p-mTOR, p-p70S6K and HIF-1α than did the control group (P < 0.05), whereas the RAP + NAM group showed slightly higher expression of these factors than did the RAP group (P < 0.05). Furthermore, in vitro studies revealed that the inhibitory effect of rapamycin on the angiogenic ability of HUVECs and its significant inhibitory effects on the protein level of HIF-1α and the phosphorylation of proteins involved in the mTORC1 pathway, including mTOR, raptor and p70S6K (P < 0.05), were enhanced by cotreatment with SRT1720 and rapamycin (P < 0.05). In contrast to mTOR and SIRT1, the mRNA levels of p70S6K and HIF-1α were reduced by rapamycin (P < 0.05) and further reduced by cotreatment with SRT1720 and rapamycin. The study results indicate that SIRT1 might negatively regulate atherosclerotic angiogenesis via mTORC1 and HIF-1α signaling pathway and cointervention of SIRT1 and mTOR may serve as a crucial therapeutic strategy in cardiovascular medicine.


Evaluation of activity of epiphyseal plates in growing males and females.
To investigate the age-related activity of the epiphyseal plates, a retrospective study of (99m)Tc-methylene diphosphonate bone scans was undertaken. The study comprised 81 males and 46 females aged 2 weeks to 24 years. The total percentage (%) whole-body (ratio of total physis activity to whole-body activity) and the regional % whole-body (ratio of physis activity of one region to whole-body activity) were derived. The ratio of physis activity of one region to the total physis activity was defined as % physis. Before age 12, total physis activity was found to contribute about 10% to whole-body activity. All total and regional % whole-body activities followed sigmoid curves with age. The differences of the parameters (transition centers and widths) suggested that there might be a later and longer period for the disappearance of physis activity in males than in females. For all the regions, % physis changed little with age until after puberty. At age <1, the proportion of bone activity in the body was about 30-35% for skull, 20-25% for lower limbs, and 5-15% for the rest of the regions. The maximal changes during growth occurred in the skull and the lower limbs. The age-related changes of physis activity during growth reflect a combination of the potential of bone to grow and the processes of bone growth and bone turnover. Bone scintigraphy is useful in understanding the changes of physis activity during growth.


The Obscure Potential of AHNAK2.
AHNAK2 is a protein discovered in 2004, with a strong association with oncogenesis in various epithelial cancers. It has a large 616 kDa tripartite structure and is thought to take part in the formation of large multi-protein complexes. High expression is found in clear cell renal carcinoma, pancreatic ductal adenocarcinoma, uveal melanoma, and lung adenocarcinoma, with a relation to poor prognosis. Little work has been done in exploring the function and relation AHNAK2 has with cancer, with early studies showing promising potential as a future biomarker and therapeutic target.


Heavy chain ferritin activates regulatory T cells by induction of changes in dendritic cells.
Heavy chain ferritin (H-ferritin) is a component of the iron-binding protein, ferritin. We have previously shown that H-ferritin inhibits anti-CD3-stimulated lymphocyte proliferation and that this was due to increased production of interleukin-10 (IL-10). In the present study we have shown that induction of IL-10 production was due to effects of H-ferritin on adherent antigen-presenting cells (APCs) in blood and monocyte-derived dendritic cells (MoDCs). IL-10 was produced by a subpopulation of CD4 T cells, which expressed the CD25 component of the IL-2 receptor and the CTLA-4 receptor characteristic of regulatory T cells. The changes induced in MoDCs were compared with those induced by CD40L and their significance tested by inhibition with monoclonal antibodies. These studies indicated that H-ferritin induced relatively greater expression of CD86 and B7-H1 on MoDCs and that monoclonal antibodies against their receptors, CTLA-4 and programmed death receptor-1 (PD-1), inhibited IL-10 production from the regulatory T cells. H-ferritin did not appear to induce direct production of the cytokines IL-2, IL-4, IL-6, IL-10, IL-12, or interferon-gamma from the DCs. These results are consistent with the thesis that H-ferritin induces B7-H1 and CD86 (B7-2) on APCs, which in turn induce IL-10 production from regulatory T cells. This is possibly one mechanism by which melanoma cells may induce changes in APCs in the vicinity of the tumor and result in suppression of immune responses by induction of regulatory T cells.


Adult-type hypolactasia and calcium intake in polycystic ovary syndrome.
Adult-type hypolactasia (ATH) is related to lower calcium and milk intake, which might be associated with obesity and metabolic disturbances. Women with polycystic ovary syndrome (PCOS) frequently suffer from metabolic disturbances including central obesity. We aimed to examine the association of ATH and calcium intake with anthropometric, metabolic and endocrine parameters in a cohort of PCOS and control women. Metabolic, endocrine and anthropometric measurements and oral glucose tolerance tests were performed in 504 PCOS and 366 control women. Genotyping of ATH, defined by the -13910 variant of the MCM6 gene, was performed. Calcium intake was assessed by questionnaires. Adult-type hypolactasia was more prevalent in PCOS women (29·8%) than in controls (23·5%) (P = 0·040). PCOS women with ATH had higher waist-to-hip ratio (WHR) (0·80 [0·75-0·88] vs 0·78 [0·73-0·85], P = 0·046), glucose 2 h (5·28 [4·57-6·33] mmol/l vs 5·67 [4·68-6·78] mmol/l, P = 0·037), HbA1c (5·2 [5·0-5·4]%vs 5·1 [5·0-5·3]%, P = 0·009), parathyroid hormone (3·72(2·91-4·86] pmol/l vs 3·61 [2·94-4·63] pmol/l, P = 0·030) and Ferriman-Gallwey-Scores (FG Scores) (7 [3-12] vs 4 [1-9], P = 0·002) and lower 25(OH)D levels (54·4 [35·2-80·6] nmol/l vs 68·4 [49·7-89·4] nmol/l, P < 0·001) than PCOS women without ATH. The association of 25(OH)D and FG-Scores with ATH remained significant in age-, BMI- and WHR-adjusted analyses. PCOS women within the highest quartile of calcium intake had significantly lower testosterone (P = 0·023) and androstenedione (P = 0·032) and significantly higher high-density lipoprotein (HDL) levels (P = 0·035) than PCOS women with lower calcium intake. Our results indicate an association of ATH with PCOS susceptibility. Moreover, ATH might influence WHR, HbA1c and FG-Scores as well as 25(OH)D levels. Higher calcium intake was associated with lower androgens and higher HDL levels.


Analysis of Cyanogenic Compounds Derived from Mandelonitrile by Ultrasound-Assisted Extraction and High-Performance Liquid Chromatography in Rosaceae and Sambucus Families.
An analytical method for extraction and quantitative determination of amygdalin, prunasin, and sambunigrin in plant material is described. The method is based on extraction with high-power ultrasound (UAE), with acidified water as solvent and quantification by HPLC-DAD. The best extraction conditions were: 80% sonication amplitude, 55 s extraction time, 70% duty cycle, 0.1 g sample mass, and 10 mL acidified water (0.1% perchloric acid). Once developed, the method was validated in terms of accuracy and precision. Good linearity was obtained, with correlation coefficients exceeding 0.999 and the quantification limits ranged from 2.2 μg/g (amygdalin) to 9.6 μg/g (sambunigrin). The accuracy (recovery study) ranged between 90 and 104% and the reproducibility of the method was always <2.3% (RSD). Special attention should be paid to the ratio sample/solvent in samples with potential β-glucosidase activity to avoid degradation of the cyanogenic glycosides (CNGs). The proposed method was used to evaluate the content of CNGs in kernels of Prunus genera, apple seeds, apple pomace, and different plant materials of Sambucus nigra.


Some Common SNPs of the T-Cell Homeostasis-Related Genes Are Associated with Multiple Sclerosis, but Not with the Clinical Manifestations of the Disease, in the Polish Population.
Multiple sclerosis (MS) is an autoimmune disease, and genetic factors play an important role in its pathogenesis and progression. The aim of our study was to evaluate the frequencies of alleles and genetic variants of the T-cell homeostasis-related genes, in subjects with MS, as well as to investigate the association with MS clinical manifestations and disability. 94 subjects with MS and 160 healthy individuals have been genotyped for seven common single-nucleotide variants in IL-2RA, CTLA4, CD40, and PADI4 genes. The ages of onset, duration of the disease, and clinical condition of the MS subjects were analysed. We used the Chi2 test confirmed with Fisher's exact test for statistical analysis. The frequency of allele T and CT/TT genotypes (rs7093069) in the IL2RA gene, as well as the T allele and CT/TT genotypes in rs12722598, were significantly higher in the control group. The significant differences between studied groups we also found for the G allele and GG/GA genotypes of rs3087243 in CTLA4 gene, which were more common among the control group. The heterozygous genotype TC (rs1883832) of CD40 gene was more common in the control subjects, and the frequency of the alleles and genotypes in the rs1748033 of the PADI4 gene did not differ between the studied groups. Between the studied genotypes, we did not observe any significant differences in the age of onset and duration of disease, including sex stratification. Our results highlight the protective role of some of the T-cell homeostasis-related genetic variants in MS development, but not in its clinical manifestation.


Visual modulation of otolith-dependent units in cat vestibular nuclei.
Vestibular nucleus units responding to translational self-motion are shown to respond also to translational movement of a large visual field. These visual-vestibular interactions in otolith-dependent units are similar to those found in canal-related units and could provide the basis for linearvection.


Superior patient and technique survival with very high standard Kt/V in quotidian home hemodialysis.
We studied the association of patient and dialysis factors with patient and technique survival in a cohort of all of our 191 of patients surviving >3 months on quotidian home hemodialysis (QHHD). Eighty-one patients were on nocturnal QHHD and 110 on short -daily QHHD. Weekly dialysis time was 7.5-48 hours, single pool Kt/V was 0.38-4.5 per treatment, and weekly standardKt/V was 2.1-7.5. The association of 18 patient and dialysis variables with patient and technique survival was analyzed by Kaplan-Meier and Cox analyses. Ninety-nine patients (52%) remained on QHHD, 34 (18%) were transplanted, 31 (16%) returned to 3/week HD, and 27 (14%) died. The 5-year patient survival was 71% ± 6% (night: 79% ± 7%, day: 69% ± 9%, P = 0.002). The 5-year technique survival was 80% ± 4% (night: 93% ± 3%, day: 46% ± 17%, P = 0.001). In Cox analyses, patient survival was independently associated with standard Kt/V (hazard ratio [HR] = 0.29, P < 0.0001), graduating from high school (HS) (HR = 0.11, P = 0.0002), and use of graft/fistula (HR = 0.22, P = 0.007). Technique survival was independently associated with standard Kt/V (HR = 0.50, P = 0.0003) and start of QHHD after 2003 (HR = 0.18, P = 0.007). Every increase in standard Kt/V was associated with improved survival. The highest survival occurred when standard Kt/V exceeded 5.1, only possible when weekly dialysis hours exceed 35 hours. In QHHD, higher standard Kt/V, education, and subcutaneous access are associated with better patient survival and higher standard Kt/V and longer experience of center with better technique survival. There was no upper limit of standard Kt/V, where survival plateaus. The amount of minimally "adequate" dialysis should be much increased.


Postnatal cumulative incidence of supraventricular tachycardia in a general pediatric population: A national birth cohort database study.
Supraventricular tachycardia (SVT) is a common pediatric tachycardia, but the true incidence is unknown. We sought to investigate the true postnatal incidence and its medical needs. We derived a birth cohort comprised children born between 2000 and 2008 who had complete postnatal medical data in the Taiwan National Health Insurance Database for the period from 2000 to 2014. From 1,967,911 live births, we identified 2021 patients with SVT (51.6% men), accounting for an overall incidence of 1.03 per 1000 patient-years (Wolff-Parkinson-White syndrome accounted for 16.2%). The cumulative incidence was 0.06, 0.25, 0.45, 0.88, and 1.39 per 1000 patient-years by the age of 1 month, 1 year, 5 years, 10 years, and 15 years, respectively. Major congenital heart disease (5.3%; hazard ratio 6.66; 95% confidence interval 2.98-14.87) and cardiomyopathy (0.9%; hazard ratio 8.78; 95% confidence interval 3.39-22.78) were associated with mortality. In patients without major congenital heart disease, the cumulative incidence of SVT was 0.05, 0.22, 0.41, 0.84, and 1.33 per 1000 patient-years by the age of 1 month, 1 year, 5 years, 10 years, and 15 years, respectively. By the age of 15 years, the annual risk of death and sudden death was 0.13% and 0.01% per patient-year, respectively. Radiofrequency catheter ablation was performed in 173 patients at the median age of 11 years: 1.7% during infancy, 5.8% by the age of 5 years, and 31.8% by the age of 10 years. The probability of being free from receiving ablation by the age of 15 years was 83.4%. This birth cohort study provides the true incidence of pediatric SVT and indicates that almost one-fifth of the patients with SVT have already received ablation in the pediatric ages.


Studies on chemically induced dominant lethality. III. Cytogenetic analyses of TEM-effects on maturing dictyate mouse oocytes.
A comparative cytogenetic study was done on metaphase-I oocytes and first cleavage mitoses following treatment of young mature female mice with triethylenemelamine (TEM). The ova were collected at intervals ranging from 12 h to 10.5 days after single intraperitoneal injection of TEM. Very few structural aberrations were seen in the metaphase-I cells after TEM treatment. There was, however, a very clear effect on the female genome when first cleavage mitoses were analyzed. The aberrations seen were principally chromatid deletions and interchanges, and their frequency varied with dose and the time between treatment and mating.


Influenza Vaccine-Induced CD4 Effectors Require Antigen Recognition at an Effector Checkpoint to Generate CD4 Lung Memory and Antibody Production.
Previously, we discovered that influenza-generated CD4 effectors must recognize cognate Ag at a defined effector checkpoint to become memory cells. Ag recognition was also required for efficient protection against lethal influenza infection. To extend these findings, we investigated if vaccine-generated effectors would have the same requirement. We compared live infection with influenza to an inactivated whole influenza vaccine. Live infection provided strong, long-lasting Ag presentation that persisted through the effector phase. It stimulated effector generation, long-lived CD4 memory generation, and robust generation of Ab-producing B cells. In contrast, immunization with an inactivated virus vaccine, even when enhanced by additional Ag-pulsed APC, presented Ag for 3 d or less and generated few CD4 memory cells or long-lived Ab-producing B cells. To test if checkpoint Ag addition would enhance this vaccine response, we immunized mice with inactivated vaccine and injected Ag-pulsed activated APC at the predicted effector checkpoint to provide Ag presentation to the effector CD4 T cells. This enhanced generation of CD4 memory, especially tissue-resident memory in the lung, long-lived bone marrow Ab-secreting cells, and influenza-specific IgG Ab. All responses increased as we increased the density of peptide Ag on the APC to high levels. This suggests that CD4 effectors induced by inactivated vaccine require high levels of cognate Ag recognition at the effector checkpoint to most efficiently become memory cells. Thus, we suggest that nonlive vaccines will need to provide high levels of Ag recognition throughout the effector checkpoint to optimize CD4 memory generation.


Bienzymatic glucose biosensor based on direct electrochemistry of cytochrome c on gold nanoparticles/polyaniline nanospheres composite.
Gold nanoparticles (GNPs) assembled polyaniline nanospheres (PANS) composite were prepared by noncovalent strategy. Cytochrome c (Cyt c) was electrostatically adsorbed on the GNPs/PANS modified electrode. The direct electron transfer (DET) between Cyt c and GNPs/PANS modified electrode was investigated. Cyt c shows a couple of quasi-reversible and well-defined cyclic voltammetry (CV) peaks with a formal potential (E(0')) of -0.27 V (versus Ag/AgCl) in pH 7.0 phosphate buffer solution (PBS). The Cyt c/GNPs/PANS modified electrode gives an improved electrocatalytic activity toward the reduction of hydrogen peroxide (H2O2). The catalysis currents increase linearly to the H2O2 concentration in a wide range of 0.01-2.4 mM with a correlation coefficient 0.998. After immobilized glucose oxidase (GOD) on the Cyt c/GNPs/PANS modified electrode by Nafion polymer, a bienzymatic glucose biosensor was further prepared for sensitive detection of glucose. The amperometric detection of glucose was assayed by potentiostating the bienzymatic electrode at -0.2 V versus Ag/AgCl to reduce the enzymatically produced H2O2 with minimal interference from the coexisting electroactive compounds. The proposed glucose biosensor exhibits good response performance to glucose with a wide linear range from 0.01 to 3.2mM with a correlation coefficient of 0.997. The electrode has high sensitivity of 63.1 μA mM(-1)cm(-2) and a detection limit 0.01 mM at the signal-to-noise ratio of 3. Moreover, the glucose biosensor possesses good stability and reproducibility.


Soil microbe-mediated carbon and nitrogen cycling during primary succession of biological soil crusts in tailings ponds.
Tailings ponds resulting from mining operations have led to serious environmental hazards, and their bioremediation is an area of ongoing exploration. Primary succession represents the starting point of biotic community establishment and development, with soil carbon and nitrogen cycling being critical to this process. To investigate the soil microbial-mediated carbon and nitrogen cycling patterns accompanying primary succession, we selected three types of tailings ponds as study areas and set up sampling sites for different stages of primary succession. The results showed that primary succession promoted microbe-mediated carbon and nitrogen cycling. It also led to improvements in soil nutrient availability and enzyme activity. In primary succession, the main pathways of carbon cycling are 3HP and rTCA, and nitrogen cycling is nitrate assimilation. In the early stages, microbes mediated more anaerobic and microaerobic processes. As succession proceeded, the pattern of microbial contributions to the carbon and nitrogen cycles changed. As succession proceeds, the functional metabolic potential of the carbon cycle gradually rises, while the nitrogen cycle shows a dramatic increase after the accumulation of autotrophic biomass. In addition, we found a positive coupling pattern between the carbon and nitrogen cycles. These findings support the optimization of bioremediation strategies for tailings ponds.


How physicians document outpatient visit notes in an electronic health record.
Clinical documentation, an essential process within electronic health records (EHRs), takes a significant amount of clinician time. How best to optimize documentation methods to deliver effective care remains unclear. We evaluated whether EHR visit note documentation method was influenced by physician or practice characteristics, and the association of physician satisfaction with an EHR notes module. We surveyed primary care physicians (PCPs) and specialists, and used EHR and provider data to perform a multinomial logistic regression of visit notes from 2008. We measured physician documentation method use and satisfaction with an EHR notes module and determined the relationship between method and physician and practice characteristics. Of 1088 physicians, 85% used a single method to document the majority of their visits. PCPs predominantly documented using templates (60%) compared to 34% of specialists, while 38% of specialists predominantly dictated. Physicians affiliated with academic medical centers (OR 1.96, CI (1.23, 3.12)), based at a hospital (OR 1.57, 95% CI (1.04, 2.36)) and using the EHR for longer (OR 1.13, 95% CI (1.03, 1.25)) were more likely to dictate than use templates. Most physicians of 383 survey responders were satisfied with the EHR notes module, regardless of their preferred documentation method. Physicians predominantly utilized a single method of visit note documentation and were satisfied with their approach, but the approaches they chose varied. Demographic characteristics were associated with preferred documentation method. Further research should focus on why variation exists, and the quality of the documentation resulting from different methods used.


[The effect of hemodilution and autotransfusion on diagnostic levels of digoxin in serum of patients operated on with extracorporeal circulation].
In many patients operated on for valvular heart diseases indications are present to digitalis administration in the preoperative period. During extracorporeal circulation fluctuations occur of blood potassium concentration which increase the risk for toxic effect of digitalis. In the work, digoxin concentration was studied in the serum of patients operated on in extracorporeal circulation in whom administration of this drug was discontinued 48 hours before the operation. The first digoxin dose (0.5 mg) was given directly after withdrawal of extracorporeal circulation, and another 0.5 mg was given after 4 hours. Digoxin concentrations were determined by fluorescence-polarization-immunological method using TD* Abbott device. The digoxin concentration increased rapidly and reached its peak after 6 hours (1.9 ng/ml), decreasing after 12 hours to 0.9 ng/ml and persisted at this therapeutic level in further samples. The present studies demonstrated that digoxin administration directly after withdrawal of extracorporeal circulation and within 24 hours after the operation in commonly accepted doses provided therapeutic concentration of the drug and caused no increase of the risk for toxic reaction development.


[The expression of Cath-D, c-erbB-2 and EGFR in breast cancer and its correlation to lymphatic metastasis].
The expression of Cath-D c-erbB-2 and EGFR in breast cancer and its correlation of lymphatic metastases were studied in 277 cases by immunohistochemical technique. Positive staining of Cath-D was detected in 107 cases (38.62%), of which, 89 cases (83.17%) had documented metastasis in the lymph nodes. One hundred and seventy cases (61.38%) stained negative for Cath-D, of which 64 cases (37.64%) had detectable lymphatic metastases. There was a significant difference in the rate of the lymphatic metastases between the Cath-D positive and Cath-D negative groups (P < 0.0001). Fifty-six out of 107 Cath-D positive cases (52.34%) were c-erbB-2 positive as well. However, only 27 out of 170 Cath-D negative cases (15.88%) were c-erbB-2 positive. The positive rate of c-erbB-2 in Cath-D positive group was significantly different from that of Cath-D negative group (P < 0.0001). Among those 107 Cath-D positive cases, 49 cases (45.79%) were EGFR positive. Only 24 cases (14.12%) were EGFR positive among the 170 Cath-D negative cases. The positive rate of EGFR between these two groups was also significantly different (P < 0.0001). The results suggest that Cath-D status can be used as an indicator of the aggressiveness of the breast cancer. Breast cancer cases with a positive Cath-D staining are more likely to have lymphatic metastases and a poor prognosis. Therefore, alternative therapeutic strategies and close follow-ups are needed for these patients.


Targeting malaria parasite proteins to the erythrocyte.
The intraerythrocytic stages of the protozoan parasite Plasmodium falciparum reside within a parasitophorous vacuole (PV) and set up unique "extraparasite, intraerythrocyte" protein-trafficking pathways that target parasite-encoded proteins to the erythrocyte cytoplasm and cell surface. Two recent articles report the identification of trafficking motifs that regulate the transport of parasite-encoded proteins across the PV. These articles greatly aid the annotation of the parasite "secretome" catalog of proteins that are targeted to the erythrocyte cytoplasm or cell membrane.


[State of the stroma in melanoma in patients with different delayed hypersensitivity reaction to neoplasm antigen].
The data are reported on studying the stromal response of 19 melanomas of analogous localization, showing the same type of growth, histological structure, the level of invasion and area of ulceration in patients with positive and negative delayed hypersensitivity reaction to injection of the polysaccharide fraction from melanoma tissue. A high degree of pronouncement of lymphoid-cellular infiltration of tumor stroma correlated with a positive delayed hypersensitivity response, whereas a low degree -- with a negative one.


Surviving verbal abuse: a win-win game plan.
Verbal abuse, one of many tactics used when communication breaks down, aggressively manipulates others into competitive power struggles loaded with hidden agendas. A specific win-win game plan is given to survive these verbal abuse tactics.


Spatial and temporal visual filtering in patients with glaucoma and ocular hypertension.
To investigate the spatial and temporal response characteristics in patients with glaucoma and ocular hypertension (OHT), in order to demonstrate the earliest specific measure of vision loss and to elucidate the precise mechanism of early glaucomatous damage. The Open Optical View System was used to assess changes in spatial and temporal vision in patients with clearly defined glaucoma (30 eyes), patients with OHT (30 eyes) and 30 eye-matched normal control eyes. The psychophysical methods applied were developed by Barber and Ruddock, who showed that the responses called ST1 and ST2 have similar characteristics to those found in the parvocellular and magnocellular pathway respectively. The measurement of spatial and temporal responses rests upon the measurement of a target that moves across a background, itself modulated spatially (gratings) and temporally (flicker). The results reported, taken together with the current data on retinal ganglion cell function, reveal damage to both the parvocellular and magnocellular pathway in patients with glaucoma. In the OHT groups only marginal changes were found in the parvocellular response, but significant changes in the magnocellular response. In both OHT and glaucoma, the temporal processing shows greatest abnormalities at a small number of low flicker frequencies. We provide psychophysical evidence of the detectable ganglion cell damage in patients with OHT who remain normal on testing with automated perimetry. The psychophysical method chosen (which measures the function of specific sub-divisions of the visual pathway) may prove useful in screening for glaucomatous damage.


Rheumatoid Arthritis Exacerbates the Severity of Osteonecrosis of the Jaws (ONJ) in Mice. A Randomized, Prospective, Controlled Animal Study.
Rheumatoid arthritis (RA), an autoimmune inflammatory disorder, results in persistent synovitis with severe bone and cartilage destruction. Bisphosphonates (BPs) are often utilized in RA patients to reduce bone destruction and manage osteoporosis. However, BPs, especially at high doses, are associated with osteonecrosis of the jaw (ONJ). Here, utilizing previously published ONJ animal models, we are exploring interactions between RA and ONJ incidence and severity. DBA1/J mice were divided into four groups: control, zoledronic acid (ZA), collagen-induced arthritis (CIA), and CIA-ZA. Animals were pretreated with vehicle or ZA. Bovine collagen II emulsified in Freund's adjuvant was injected to induce arthritis (CIA) and the mandibular molar crowns were drilled to induce periapical disease. Vehicle or ZA treatment continued for 8 weeks. ONJ indices were measured by micro-CT (µCT) and histological examination of maxillae and mandibles. Arthritis development was assessed by visual scoring of paw swelling, and by µCT and histology of interphalangeal and knee joints. Maxillae and mandibles of control and CIA mice showed bone loss, periodontal ligament (PDL) space widening, lamina dura loss, and cortex thinning. ZA prevented these changes in both ZA and CIA-ZA groups. Epithelial to alveolar crest distance was increased in the control and CIA mice. This distance was preserved in ZA and CIA-ZA animals. Empty osteocytic lacunae and areas of osteonecrosis were present in ZA and CIA-ZA but more extensively in CIA-ZA animals, indicating more severe ONJ. CIA and CIA-ZA groups developed severe arthritis in the paws and knees. Interphalangeal and knee joints of CIA mice showed advanced bone destruction with cortical erosions and trabecular bone loss, and ZA treatment reduced these effects. Importantly, no osteonecrosis was noted adjacent to areas of articular inflammation in CIA-ZA mice. Our data suggest that ONJ burden was more pronounced in ZA treated CIA mice and that RA could be a risk factor for ONJ development. © 2016 American Society for Bone and Mineral Research.


Low serum sclerostin levels in newborns with vitamin D deficiency.
Sclerostin is a glycoprotein produced by osteocytes that is being evaluated as a potential clinical marker of bone turnover. The aim of this study was to investigate the association between neonatal vitamin D status and levels of circulating sclerostin. Forty newborns were recruited for the study. Vitamin D deficiency was defined as a serum 25-hydroxyvitamin D3 [25(OH)D] concentration <20 ng/mL and the newborns were divided into two groups as vitamin D deficient and vitamin D sufficient groups. Calcium, phosphate, alkaline phosphatase and sclerostin were measured at birth. Newborns with vitamin D deficiency had markedly lower 25(OH)D levels than vitamin D sufficient newborns (8.5±4.4 ng/mL vs. 35.3±10.6 ng/mL, p<0.001). Vitamin D deficient infants also had significantly lower serum sclerostin levels (188.4±21.9 vs. 282.3±30.4 pg/mL; p: 0.026) than vitamin D sufficient newborns at birth. However, we did not detect a significant linear association between neonatal sclerostin and maternal/neonatal 25(OH)D levels. Our data also demonstrated that vitamin D deficient newborns exhibited lower sclerostin levels than vitamin D sufficient newborns. The low sclerostin level might serve as a marker of decreased osteocyte activity in newborns with vitamin D deficiency.


General linear models.
This chapter presents the general linear model as an extension to the two-sample t-test, analysis of variance (ANOVA), and linear regression. We illustrate the general linear model using two-way ANOVA as a prime example. The underlying principle of ANOVA, which is based on the decomposition of the value of an observed variable into grand mean, group effect and random noise, is emphasized. Further into this chapter, the F test is introduced as a means to test for the strength of group effect. The procedure of F test for identifying a parsimonious set of factors in explaining an outcome of interest is also described.


Somatotopic organization and functional properties of mechanosensory neurons expressing sensorin-A mRNA in Aplysia californica.
A previous study reported that a peptide, sensorin-A, is expressed exclusively in mechanosensory neurons having somata in central ganglia of Aplysia. The present study utilized in situ hybridization, staining by nerve back-fill and soma injection, and electrophysiological methods to characterize the locations, numbers, and functions of sensorin-A-expressing neurons and to define the relationships between soma locations and the locations of peripheral axons and receptive fields. Approximately 1,000 cells express sensorin-A mRNA in young adult animals (10-30 g) and 1,200 cells in larger adults (100-300 g). All of the labeled somata are in the CNS, primarily in the abdominal LE, rLE, RE and RF, pleural VC, cerebral J and K, and buccal S clusters. Expression also occurs in a few sparsely distributed cells in most ganglia. Together, receptive fields of all these mechanosensory clusters cover the entire body surface. Each VC cluster forms a somatotopic map of the ipsilateral body, a "sensory aplunculus." Cells in the pleural and cerebral clusters have partially overlapping sensory fields and synaptic targets. Buccal S cells have receptive fields on the buccal mass and lips and display notable differences in electrophysiological properties from other sensorin-A-expressing neurons. Neurons in all of the clusters have relatively high mechanosensory thresholds, responding preferentially to threatening or noxious stimuli. Synaptic outputs to target cells having defensive functions support a nociceptive role, as does peripheral sensitization following noxious stimulation, although additional functions are likely in some clusters. Interesting questions arise from observations that mRNA for sensorin-A is present not only in the somata but also in synaptic regions, connectives, and peripheral fibers.


European viper envenomation recorded by French poison control centers: A clinical assessment and management study.
Immunotherapy is the gold standard treatment for patients bitten by European vipers in France; it significantly decreases morbidity, frequency and severity of complications and length of stay. A national prospective study was performed by all Poison Control Centers (PCC) to validate the emergency protocol for viper envenomations. This prospective study included all cases of viper bites in France, treated or not with Viperfav(®) in 2013. In 2013, 277 cases of viper bites were collected: ratio M/F 2.1; mean aged 43 years (<15 years 25% 15-65 63% > 65 12%). The final severity was divided into 68 grades 0, 58 grades I, 62 grades IIA, 71 grades IIB and 18 grades III. One death was reported. Five patients had neurological signs. For the 114 patients who received Viperfav(®), all systemic signs disappeared in 5 h and in 24 h for biological and neurological signs. No severe anaphylactic reaction with Viperfav(®) was reported. Late Viperfav(®) administration increased the risk of functional impairment 15 days after the bite (OR = 3.21 p = 0.043). The administration of Low Molecular Weight Heparin (LMWH) increased the frequency of functional impairment to 15 days after the bite (OR = 6.38 p = 0.064), although Viperfav(®) was given in the first 18 h. This study confirms the efficiency, safety and recommendation of an early administration of a single dose of Viperfav(®), LMWH should not be used. It also shows the extension of neurotoxic venom of vipers in France.


Multistage microfluidic cell sorting method and chip based on size and stiffness.
High performance sorting of circulating tumor cells (CTCs) from peripheral blood is key to liquid biopsies. Size-based deterministic lateral displacement (DLD) technique is widely used in cell sorting. But conventional microcolumns have poor fluid regulation ability, which limits the sorting performance of DLD. When the size difference between CTCs and leukocytes is small (e.g., less than 3 μm), not only DLD, many size-based separation techniques fail due to low specificity. CTCs have been confirmed to be softer than leukocytes, which could serve as a basis for sorting. In this study, we presented a multistage microfluidic CTCs sorting method, first sorting CTCs using a size-based two-array DLD chip, then purifying CTCs mixed by leukocytes using a stiffness-based cone channel chip, and finally identifying cell types using Raman techniques. The entire CTCs sorting and analysis process was label free, highly pure, high-throughput and efficient. The two-array DLD chip employed a droplet-shaped microcolumn (DMC) developed by optimization design rather than empirical design. Attributed to the excellent fluid regulation capability of DMC, the CTCs sorter system developed by parallelizing four DMC two-array DLD chips was able to process a sample of 2.5 mL per minute with a recovery efficiency of 96.30 ± 2.10% and a purity of 98.25 ± 2.48%. To isolate CTCs mixed dimensionally by leukocytes, a cone channel sorting method and chip were developed based on solid and hydrodynamic coupled analysis. The cone channel chip allowed CTCs to pass through the channel and entrap leukocytes, improving the purity of CTCs mixed by leukocytes by 1.8-fold.


Altered mRNP granule dynamics in FTLD pathogenesis.
In neurons, RNA-binding proteins (RBPs) play a key role in post-transcriptional gene regulation, for example alternative splicing, mRNA localization in neurites and local translation upon synaptic stimulation. There is increasing evidence that defective or mislocalized RBPs - and consequently altered mRNA processing - lead to neuronal dysfunction and cause neurodegeneration, including frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Cytosolic RBP aggregates containing TAR DNA-binding protein of 43 kDa (TDP-43) or fused in sarcoma (FUS) are a common hallmark of both disorders. There is mounting evidence that translationally silent mRNP granules, such as stress granules or transport granules, play an important role in the formation of these RBP aggregates. These granules are thought to be 'catalytic convertors' of RBP aggregation by providing a high local concentration of RBPs. As recently shown in vitro, RBPs that contain a so-called low-complexity domain start to 'solidify' and eventually aggregate at high protein concentrations. The same may happen in mRNP granules in vivo, leading to 'solidified' granules that lose their dynamic properties and ability to fulfill their physiological functions. This may result in a disturbed stress response, altered mRNA transport and local translation, and formation of pathological TDP-43 or FUS aggregates, all of which may contribute to neuronal dysfunction and neurodegeneration. Here, we discuss the general functional properties of these mRNP granules, how their dynamics may be disrupted in frontotemporal lobar degeneration/amyotrophic lateral sclerosis, for example by loss or gain of function of TDP-43 and FUS, and how this may contribute to the development of RBP aggregates and neurotoxicity. In this review, we discuss how dynamic mRNP granules, such as stress granules or neuronal transport granules, may be converted into pathological aggregates containing misfolded RNA-binding proteins (RBPs), such as TDP-43 and FUS. Abnormal interactions between low-complexity domains in RBPs may cause dynamic mRNP granules to solidify and become dysfunctional. This may result in a disturbed stress response, altered mRNA transport and local translation, as well as RBP aggregation, all of which may contribute to neuronal dysfunction and neurodegeneration.


Metal-Ceramic Compatibility in Dental Restorations According to the Metallic Component Manufacturing Procedure.
In terms of production technology, metal-ceramic systems for dental restorations comply with a concrete algorithm, the efficiency of which is always dependent on the applications for which they are intended. The first stage involves obtaining metal support, followed by firing the ceramic on the surface of the metal to meet the list of functional and aesthetic requirements of a future restoration. The compatibility of the two materials-the metal component and the ceramic component-must be ensured in several respects: chemical compatibility, thermo-chemical compatibility, and mechanical compatibility. Thus, there is a need to simulate the thermal behavior of the metal-ceramic couple in its processing to achieve appropriate dental prostheses. In this study, three types of Co-Cr metal frames were manufactured using three different production technologies: conventional casting, milling (CAM), and selective laser melting (SLM). Composition analyses, scanning electron microscopy (SEM), and microstructural analyses of the metal-ceramic interface for each type of production technology, as well as the determination of the hardness and the thermal expansion coefficients of experimental materials and three-point bending tests, were carried out in this study. Considering all these aspects, we demonstrated the influence of the technology of producing the metallic part of the metal-ceramic bonding process in dental prostheses.


Correction: Analyzing data from the digital healthcare exchange platform for surveillance of antibiotic prescriptions in primary care in urban Kenya: A mixed-methods study.
[This corrects the article DOI: 10.1371/journal.pone.0222651.].


[Reconstruction [correction to Reconstrction] of mandibular bone defects by using the techniques [correction to teachniques] of the reverse engineering and rapid prototyping].
To evaluate the feasibility of designing and fabricating customized titanium bone substitutes to restore mandibular bone defects using reverse engineering (RE) and rapid prototyping (RP) techniques. Titanium tray for mandibular defects were designed and fabricated through multi-step procedures of reverse engineering and rapid prototyping, then in operation it was filled with cancellous bone and fixed. The bone substitutes fabricated by this method had been successfully put into clinical use for maxillofacial surgery in 2 patients and got a satisfactory result. Reverse engineering combining with rapid prototyping could accomplish the design and manufacture of implant for the restoration of mandibular bone defects.


Conceptualizing race in research.
The use of race as a variable in research continues to spark debate about whether it should be used, as well as the implications it has for research on health differences. Given this continued controversy, we examined how investigators interpret the concept of "race" and whether their views of race are reflected in their published work. Thirty-three semistructured interviews were conducted with investigators from 3 southeastern universities to discuss recruitment of participants, the use of race as a variable in research and analyses, and their assessment of the National Institutes of Health mandate on the inclusion of women and minorities. The interview data were analyzed using the principles of constant comparative method, theme identification and pattern investigation. Up to 2 publications for each respondent were also used to assess the use of race in their research. Results reflect a spectrum of views on the definition of race, from biological to social. Findings also suggest that investigators think critically about the use and implications of using race in their research, although this is not consistently reflected in their published work. In our view, authors, journal editors and peer reviewers have an important role in moving this debate forward, and advocate that they engage more directly in shaping the process. When reporting results by race, investigators should provide a statement on the theory or conceptual framework underlying the hypothesized racial differences in health examined in the study. They should be also cautious in invoking either biological or social constructions of race, thus demonstrating an appreciation of the nuances and implications of using this variable.


Pure primary ovarian squamous cell carcinoma.
A 65-year-old woman with pure primary ovarian squamous cell carcinoma invading the fallopian tube and the terminal ileum is described. The tumor recurred shortly after surgery in spite of complete resection. No response to chemotherapy was observed and the patient expired 6 months after diagnosis. In order to establish the diagnosis of pure primary ovarian squamous cell carcinoma extragenital and genital tumors containing squamous elements as well as endometriosis have to be excluded.


The highest prevalence of human metapneumovirus in Ahwaz children accompanied by acute respiratory infections.
The newly described human metapneumovirus (hMPV) has been recently discovered as an etiological agent of acute respiratory infections (ARTI) in infants and children. The aim of this study was to determine the prevalence of hMPV and its potential role as causative agent of ARTI in Ahwaz children. In the present study, we examined 124 nasal swabs from infants affected by ARTI for the presence of hMPV by RT-PCR technique. Sixty-eight out of 124 (54.4%) cases were positive for hMPV which is the highest incidence of hMPV ever reported in the world, 94.1% of positive cases belonged to genotype A; whereas no B genotype was detected. Our positive hMPV children were affected by upper (URTI) as well as lower respiratory tract infection (LRTI); however, LARTIs had higher prevalence. We suggest a probable role of F protein alteration as the causative agent for the highest prevalence of hMPV infection among Ahvaz children.


Combined treatment of high-intensity interval training with neural stem cell generation on contusive model of spinal cord injury in rats.
Spinal cord injury (SCI) leads to inflammation, axonal degeneration, and gliosis. A combined treatment of exercise and neural stem cells (NSC) has been proposed to improve neural repair. This study evaluated a combined treatment of high-intensity interval training (HIIT) with NSC generation from adipose-derived stem cells (ADSCs) on a contusive model of SCI in rats. In vitro, rat ADSCs were isolated from the perinephric regions of Sprague-Dawley rats using enzymatic digestion. The ADSCs were transdifferentiated into neurospheres using B27, EGF, and bFGF. After production of NSC, they were labeled using green fluorescent protein (GFP). For the in vivo study, rats were divided into eight groups: control group, sham operation group, sham operation + HIIT group, sham operation + NSC group, SCI group, SCI + HIIT group, SCI + NSC group, and SCI/HIIT/NSC group. Laminectomy was carried out at the T12 level using the impactor system. HIIT was performed three times per week. To assess behavioral function, the Basso-Beattie-Bresnahan (BBB) locomotor test and H-reflex was carried out once a week for 12 weeks. We examined glial fibrillary acidic protein (GFAP), S100β, and NF200 expression. NSC transplantation, HIIT and combined therapy with NSC transplantation, and the HIIT protocol improved locomotor function with decreased maximum H to maximum M reflexes (H/M ratio) and increased the Basso-Beattie-Bresnahan score. Combined therapy in contused rats using the HIIT protocol and neurosphere-derived NSC transplantation improves functional and histological outcomes.


FAN score comprising fibrosis-4 index, albumin-bilirubin score and neutrophil-lymphocyte ratio is a prognostic marker of urothelial carcinoma patients treated with pembrolizumab.
It is important to identify prognostic and predictive markers of metastatic urothelial carcinoma (mUC) treated with immunocheckpoint inhibitors. We sought to establish a prognostic marker for patients with mUC treated with pembrolizumab based on only blood test results. We included 165 patients with mUC in the discovery cohort and 103 with mUC who were treated with pembrolizumab in the validation cohort. Multivariate and Cox regression analyses were used to analyse the data. In the discovery cohort, the fibrosis-4 index (hazard ratio [HR]: 2.13, 95% confidence interval [CI] 1.20-3.76, p = 0.010), albumin-bilirubin score (HR 1.91, 95% CI 1.27-2.88, p = 0.002), and neutrophil-lymphocyte ratio (HR: 1.84, 95% CI 1.22-2.79, p = 0.004) were independent significant prognostic factors. We established a 'FAN score' that included these three aforementioned items, which were assigned one point each. We divided patients into the 0-1 point (n = 116) and 2-3 points (n = 49) groups. The FAN score was a significant prognostic marker for cancer-specific survival (CSS) (HR 1.48, 95% CI 1.19-1.83, p < 0.001) along with the Eastern Cooperative Oncology Group Performance Status. The FAN score was also a prognostic factor of progression-free survival (PFS) (HR: 1.25, 95% CI 1.01-1.54, p = 0.036) along with the presence of liver metastasis. In the validation cohort, the FAN score was a significant prognostic factor for CSS (HR: 1.48, 95% CI 1.19-1.85, p = 0.001) and PFS (HR: 1.29, 95% CI 1.02-1.62, p = 0.034). We established the FAN score as a prognostic marker for patients with mUC treated with pembrolizumab.


Identification and functional prediction of stress responsive AP2/ERF transcription factors in Brassica napus by genome-wide analysis.
Using homology and domain authentication, 321 putative AP2/ERF transcription factors were identified in Brassica napus, called BnAP2/ERF TFs. BnAP2/ERF TFs were classified into five major subfamilies, including DREB, ERF, AP2, RAV, and BnSoloist. This classification is based on phylogenetic analysis, motif identification, gene structure analysis, and physiochemical characterization. These TFs were annotated based on phylogenetic relationship with Brassica rapa. BnAP2/ERF TFs were located on 19 chromosomes of B. napus. Orthologs and paralogs were identified using synteny-based methods Ks calculation within B. napus genome and between B. napus with other species such as B. rapa, Brassica oleracea, and Arabidopsis thaliana indicated that BnAP2/ERF TFs were formed through duplication events occurred before B. napus formation. Kn/Ks values were between 0 and 1, suggesting the purifying selection among BnAP2/ERF TFs. Gene ontology annotation, cis-regulatory elements and functional interaction networks suggested that BnAP2/ERF TFs participate in response to stressors, including drought, high salinity, heat and cold as well as developmental processes particularly organ specification and embryogenesis. The identified cis-regulatory elements in the upstream of BnAP2/ERF TFs were responsive to abscisic acid. Analysis of the expression data derived from Illumina Hiseq 2000 RNA sequencing revealed that BnAP2/ERF genes were highly expressed in the roots comparing to flower buds, leaves, and stems. Also, the ERF subfamily was over-expressed under salt and fungal treatments. BnERF039 and BnERF245 are candidates for salt-tolerant B. napus. BnERF253-256 and BnERF260-277 are potential cytokinin response factors. BnERF227, BnERF228, BnERF234, BnERF134, BnERF132, BnERF176, and BnERF235 were suggested for resistance against Leptosphaeria maculan and Leptosphaeria biglobosa.


Amniotic basement membrane: a barrier to neutrophil invasion.
Polymorphonuclear neutrophils activated by N-formylmethionyl-leucylphenylalanine are able to cross the human amnion as shown by Russo et al. (1981). With the same technique, we added polymorphonuclear neutrophils to either the epithelial or stromal surface of the amnion. The lower compartment of the incubation chamber contained 10(-8) mol/L of N-formylmethionyl-leucylphenylalanine. Of 24 amnion tested, only one showed polymorphonuclear neutrophil permeation. Ultrastructural observations indicated that the basal lamina and the zona reticularis of the basement membrane acted as barriers to polymorphonuclear neutrophil invasion. Recent studies indicate that the basal lamina and the zona reticularis of the basement membrane contain collagen type V. No data support the existence of collagenase of polymorphonuclear neutrophil origin that cleaves collagen type V. If enzymatic degradation of extracellular matrix components play a role in polymorphonuclear neutrophil invasion, it is possible that type V collagen acts as a barrier to polymorphonuclear neutrophil invasion in the human amnion.


The BTB-TAZ protein MdBT2 negatively regulates the drought stress response by interacting with the transcription factor MdNAC143 in apple.
Drought stress is a severe source of abiotic stress that can affect apple yield and quality, yet the underlying molecular mechanism of the drought stress response and the role of MdBT2 in the process remain unclear. Here, we find that MdBT2 negatively regulates the drought stress response. Both in vivo and in vitro assays indicated that MdBT2 interacted physically with and ubiquitinated MdNAC143, a member of the NAC TF family that is a positive regulator under drought stress. In addition, MdBT2 promotes the degradation of MdNAC143 via the 26S proteasome system. A series of transgenic assays in apple calli and Arabidopsis verify that MdBT2 confers susceptibility to drought stress at least in part by the regulation of MdNAC143. Overall, our findings provide new insight into the mechanism of MdBT2, which functions antagonistically to MdNAC143 in regulating drought stress by regulating the potential downstream target protein MdNAC143 for proteasomal degradation in apple.


A peroxyoxalate chemiluminescence-based assay for the evaluation of hydrogen peroxide scavenging activity employing 9,10-diphenylanthracene as the fluorophore.
A simple, rapid, sensitive, and enzyme-free analytical method for estimating scavenging of hydrogen peroxide (H2O2) was developed. Peroxyoxalate chemiluminescence (POCL) was measured, using 9,10-diphenylanthracene as fluorophore. The chemiluminescence signal was found to be linear in response to increasing amounts of H2O2 in ethyl acetate/acetonitrile (9:1) (r2 = .9990), within a range of concentrations varying from 9.0 to 72.0 microM. In contrast, acetonitrile was highly unsuitable because of poor linearity (r2 = .3736) and poor signal stability. The linearity of POCL inhibition, as a measure of H2O2 scavenging, was tested employing well-known, lipid-soluble antioxidants, including beta-carotene, butylated hydroxytoluene (BHT) and alpha-tocopherol, and also the more polar flavonol quercetin, and the water-soluble L-ascorbic acid (AA). Under the experimental conditions, the corresponding values of H2O2 scavenging activity (SA(HP)) for quercetin, beta-carotene, alpha-tocopherol, and L-AA were 19.1 +/- 0.4, 70.9 +/- 20.1, 8.4 +/- 0.4, and 44.8 +/- 5.6 x 10(-3) microM(-1) The data establish the assay as a method for assessing the H2O2 quenching activity of lipid-soluble antioxidants.


Auxin-regulated changes in protein phosphorylation in pea epicotyls.
Auxins regulate various aspects of plant growth and development. However, the mechanism by which these hormones elicit diverse physiological processes is not clear. We present evidence for the role of auxin in protein phosphorylation and the possible involvement of calmodulin in auxin-induced changes. In the presence of auxin, phosphorylation of 23,000, 82,000, 105,000 and 110,000 molecular weight polypeptides markedly decreased whereas phosphorylation of 19,000, 24,000 and 28,000 molecular weight polypeptides increased. These results open up a new experimental approach in understanding the molecular mechanism by which auxins regulate various physiological processes in plants.


Involvement of cannabinoid receptors and neuroinflammation in early sepsis: Implications for posttraumatic stress disorder.
Sepsis is associated with several comorbidities in survivors, such as posttraumatic stress disorder (PTSD). This study investigated whether rats that survive sepsis develop the generalization of fear memory as a model of PTSD. Responses to interventions that target the endothelin-1 (ET-1)/cannabinoid system and glial activation in the initial stages of sepsis were evaluated. As a control, we evaluated hyperalgesia before fear conditioning. Sepsis was induced by cecal ligation and puncture (CLP) in Wistar rats. CLP-induced sepsis with one or three punctures resulted in fear generalization in the survivors 13 and 20 days after the CLP procedure, a process that was not associated with hyperalgesia. Septic animals were intracerebroventricularly treated with vehicle, the endothelin receptor A (ETA) antagonist BQ123, the cannabinoid CB1 and CB2 receptor antagonists AM251 and AM630, respectively, and the glial blocker minocycline 4 h after CLP. The blockade of either CB1 or ETA receptors increased the survival rate, but only the former reversed fear memory generalization. The endothelinergic system blockade is important for improving survival but not for fear memory. Treatment with the CB2 receptor antagonist or minocycline also reversed the generalization of fear memory but did not increase the survival rate that was associated with CLP. Minocycline treatment also reduced tumor necrosis factor-α levels in the hippocampus suggesting that neuroinflammation is important for the generalization of fear memory induced by CLP. The influence of CLP on the generalization of fear memory was not related to Arc protein expression, a regulator of synaptic plasticity, in the dorsal hippocampus.


C-Jun modulates apoptosis but not terminal cell differentiation in murine erythroleukemia cells.
The proto-oncogene c-Jun has been implicated in the control of cell proliferation and differentiation and more recently in the regulation of apoptosis. We have previously reported the involvement of c-Jun in the erythroid differentiation block in murine erythroleukemia (MEL) cells. As reported here, we investigated the role of c-Jun in the regulation of terminal differentiation and apoptosis of MEL cells by studying different stable transfectant clones containing c-jun constructs in sense or antisense orientation. c-Jun did not prevent cell growth arrest in G0/G1 and p21 induction that are normally associated with terminal differentiation induced by DMSO treatment, suggesting that c-Jun may uncouple phenotypic differentiation and terminal cell division in the MEL cell system. Spontaneous apoptosis was accelerated in c-jun expressing MEL cells before and after DMSO treatment. Moreover, c-Jun sensitized apoptosis induced by various drugs. Drug-induced apoptosis was associated with c-Jun N-terminal kinase (JNK) activation and c-Jun N-terminal phosphorylation (JNP). In contrast, overexpression of c-jun delayed apoptosis in serum-starved cells, indicating that c-Jun may reduce or accelerate apoptosis in MEL cells depending on the nature of the apoptotic stimulus. These results suggest that the proto-oncogene c-Jun may modulate differentiation and apoptosis of leukemic cells.


Reconstruction of postleishmaniasis scar with lateral cheek rotation flap.
the aim of this study was to find a simple and economical technique to remove postleishmaniasis scar from the face. a non-randomized descriptive case series. the study was conducted between 1st January 1998 to 31st July 2000 in a private hospital of D.I.Khan. Ten patients with postleishmaniasis scar on the face were selected for excision and reconstruction in this study. They were operated as day care under local anesthesia. Their age ranged between 18-30 years, with an average of 23.5 years. The size of scar varied from 2 cm2 to 6 cm2 with an average of 4.1 cm(2). After scar excision, lateral cheek rotation flap (reverse face-lift) was used to reconstruct the defect. Results were statistically analyzed. No major complications were encountered. However, minor complications like edema of the flap, seroma under the flap, flap tip necrosis and temporary neurapraxia were noted in two patients. This is an excellent technique, which can be done on a day care basis under local anesthesia. Larger studies will help to explore this technique further.


Green Highly Clay-Filled Polyethylene Composites as Coating Materials for Cable Industry-A New Application Route of Non-Organophilised Natural Montmorillonites in Polymeric Materials.
In order to develop flame retardant and relatively green cable coating materials, polyethylene (PE) was melt blended with 5, 7.5, or 10 wt. % of a natural calcium montmorillonite (C-Ca) pre-dispersed in EBA (ethylene-butyl acrylate copolymer), EVA (ethylene-vinyl acetate copolymer), or mEVA (EVA modified with maleic anhydride). For comparison, an organophilised montmorillonite (CW9) was tested. The main study of composites containing EBA/C-Ca, EVA/C-Ca, and mEVA/CW9 pre-dispersions revealed that both clays were not fully exfoliated in the matrix, however, C-Ca (7.5 wt. %) markedly increased limited oxygen index (LOI) from 18% O2 (PE) up to 22.0% O2. An insignificantly higher LOI value (22.2% O2) was noted for a sample with 10 wt. % of CW9. The fillers did not affect hardness, but spectacularly increased Young's modulus of the compression-moulded samples (tensile strength and elongation at break values were reduced). Thermal features of the matrix were not unpredictably changed by the clays. Generally, all the tests revealed that PE filled with the chemically untreated natural C-Ca reached similar or better mechanical and thermal features than materials containing the ammonium salt-modified montmorillonite.