Prevalence of sickle cell disorders and malaria infection in children aged 1-12 years in the Volta Region, Ghana: a community-based study.
Alterations in the structure of haemoglobin (Hb) are usually brought about by point mutations affecting one or, in some cases, two codons encoding amino acids of the globin chains. One in three Ghanaians are said to have sickle cell disorders, whereas malaria continues to be one of the leading causes of mortality among children. This study determined the prevalence of sickle cell disorders and malaria infection among children aged 1-12 years in the Volta Region. This was a community-based cross-sectional survey that involved 938 children aged 1-12 years selected from three districts, one each from the 3 geographical zones of the Volta Region using a multistage sampling method. Demographic information was collected using a standard questionnaire and anthropometric indices were measured. Isoelectric focusing (IEF) electrophoresis was used to determine the Hb genotypes and sub-microscopic parasites were determined by PCR. The prevalence of sickling screening positive was 16.0% with an overall prevalence of sickle cell disorders being 2.0%. Among the individual genotypes making up the sickle cell disorders, genotype HbSF was the highest (0.9% as compared to 0.2%; HbSS, 0.6%; HbSC and 0.3%; HbSCF). Microscopic Plasmodium falciparum parasitaemia was detected among 5.5% of the children and 14.2% sub-microscopic prevalence by PCR. Children with sickle cell disorders were more likely to have sub-microscopic parasitaemia (AOR = 5.51 95%CI (2.15, 14.10), p < 0.001) as well as anaemia (AOR = 3.03 95% CI (1.04, 8.82), p = 0.042), compared to those with normal genotypes. There was no significant difference observed between sickle cell disorders and growth and development of the children screened. Sickle cell disorders were significantly associated with sub-microscopic parasitaemia as well as anaemia in this study. Establishment of sickle cell clinics in the district and regional hospitals will help in the management of children with the disorder and also generate a national database on sickle cell disorders. National neonatal screening policies must also be put in place to help in early detection and management of these disorders.


On the lack of coordination between protein folding and flavin insertion in Escherichia coli for flavocytochrome b2 mutant forms Y254L and D282N.
Wild-type flavocytochrome b2 (L-lactate dehydrogenase) from Saccharomyces cerevisiae, as well as a number of its point mutants, can be expressed to a reasonable level as recombinant proteins in Escherichia coli (20-25 mg per liter culture) with a full complement of prosthetic groups. At the same expression level, active-site mutants Y254L and D282N, on the other hand, were obtained with an FMN/heme ratio significantly less than unity, which could not be raised by addition of free FMN. Evidence is provided that the flavin deficit is due to incomplete prosthetic group incorporation during biosynthesis. Flavin-free and holo-forms for both mutants could be separated on a Blue-Trisacryl M column. The far-UV CD spectra of the two forms of each mutant protein were very similar to one another and to that of the wild-type enzyme, suggesting the existence of only local conformational differences between the active holo-enzymes and the nonreconstitutable flavin-free forms. Selective proteolysis with chymotrypsin attacked the same bond for the two mutant holo-enzymes as in the wild-type one, in the protease-sensitive loop. In contrast, for the flavin-free forms of both mutants, cleavage occurred at more than a single bond. Identification of the cleaved bonds suggested that the structural differences between the mutant flavin-free and holo-forms are located mostly at the C-terminal end of the barrel, which carries the prosthetic group and the active site. Altogether, these findings suggest that the two mutations induce an alteration of the protein-folding process during biosynthesis in E. coli; as a result, the synchrony between folding and flavin insertion is lost. Finally, a preliminary kinetic characterization of the mutant holo-forms showed the Km value for lactate to be little affected; kcat values fell by a factor of about 70 for the D282N mutant and of more than 500 for the Y254L mutant, compared to the wild-type enzyme.


Two novel steroids from Euphorbia officinarum latex.
Two novel steroids, 3beta,7alpha-dihydroxy-4alpha,14alpha-dimethyl-5alpha-cholest-8-en-11-one (2) and 3beta,7beta-dihydroxy-4alpha,14alpha-dimethyl-5alpha-cholest-8-en-11-one (3) were isolated from the latex of Euphorbia officinarum. Their structures were established on the basis of NMR and MS studies.


Upregulation of glycolytic enzyme PFKFB3 by deubiquitinase OTUD4 promotes cardiac fibrosis post myocardial infarction.
Metabolic dysregulations have emerged as a major mediator of cardiovascular disorders and fibrotic diseases. Metabolic reprogramming contributes a lot to cardiac fibroblast activation and cardiac fibrosis post-myocardial infarction (MI), yet the mechanism remains incompletely understood. Our work aimed to determine whether or not glycolytic reprogramming, regulated by phosphofructokinase-2/fructose-2,6-bisphosphatase 3 (PFKFB3), is a therapeutic target for alleviating post-MI cardiac fibrosis. Here, we showed that cardiac fibroblasts displayed cell energy phenotype toward augmented glycolysis in response to transforming growth factor-beta 1 (TGF-β1), evidenced by significant extracellular acidification rate (ECAR) increase and lactate accumulation. The expression of glycolytic enzyme PFKFB3, a master activator of glycolysis, was up-regulated in TGF-β1-treated cardiac fibroblasts and in cardiac fibroblasts of post-MI mice. Pharmacological inhibition of PFKFB3 by 3PO diminished TGF-β1-mediated profibrotic phenotypes, attenuated cardiac fibrosis, and preserved cardiac functions in post-MI mice. Meanwhile, the genetic inhibition of PFKFB3 decreased the cardiac fibroblast activation and reversed the differentiated phenotypes in vitro and in vivo. Mechanistically, we identified deubiquitinase OTUD4 as a new binding protein of PFKFB3, and their interaction blocked PFKFB3 degradation via OTUD4-mediated deubiquitylation. Taken together, this work characterized a key role for PFKFB3 in cardiac fibroblast activation and suggested that inhibiting PFKFB3-involved glycolysis is an alternative way to alleviate post-MI cardiac fibrosis. KEY MESSAGES: PFKFB3, a master activator of glycolysis, was highly expressed in ischemic cardiac fibroblasts to enhance cardiac fibrosis The deubiquitinase OTUD4 was identified as a new binding protein of PFKFB3 TGF-β1 blunted the ubiquitination-mediated degradation of PFKFB3 via OTUD4-mediated deubiquitylation Blockade of PFKFB3 contributed to ameliorating ischemia-induced cardiac fibrosis.


Investigation of the release behavior of cupric ion for three types of Cu-IUDs and indomethacin for medicated Cu-IUD in simulated uterine fluid.
A number of copper-bearing intrauterine devices (Cu-IUDs) are now commercially available in China. The release behavior of cupric ion from Cu-IUDs is essential to the success of contraception, and the release behavior of indomethacin from medicated Cu-IUD is related to its therapeutic effect. In this study, analytical methods were established to investigate the release behavior of cupric ion of three existing types of Cu-IUDs and indomethacin of one medicated Cu-IUD (Yuangong 365 Cu-IUD). Cu-IUDs were incubated in simulated uterine fluid (SUF). The concentrations of cupric ion and indomethacin were analyzed by flame atomic absorption spectrometer (FAAS) for 250 days and UV 752 spectrophotometer for 300 days, respectively. The release behavior of cupric ion for three types of Cu-IUDs was biphasic, which consisted of the initial burst release and then slow and constant release. The release of cupric ion from the medicated Cu-IUD Yuangong 365 showed a zero-order process. In vitro release experiment confirmed a sustained release of indomethacin from Yuangong 365 and the release was in accordance with the Weibull equation. The cupric ion release appears to be more constant in the medicated Cu-IUD than in the others. In view of the feature of indomethacin release from medicated Cu-IUD, it is conjectured that the design of this device could be useful to avoid the adverse effects caused by all Cu-IUDs.


Overcoming the challenges of interpreting complex and uncommon RH alleles from whole genomes.
Rh is one of the most diverse and complex blood group systems. Recently, next generation sequencing (NGS) has proven to be a viable option for RH genotyping. We have developed automated software (bloodTyper) for determining alleles encoding RBC antigens from NGS-based whole genome sequencing (WGS). The bloodTyper algorithm has not yet been optimized and evaluated for complex and uncommon RH alleles. Twenty-two samples with previous polymerase chain reaction (PCR) and Sanger sequencing-based RH genotyping underwent WGS. bloodTyper was used to detect RH alleles including those defined by structural variation (SV) using a combination of three independent strategies: sequence read depth of coverage, split reads and paired reads. bloodTyper was programmed to identify D negative and positive phenotypes as well as the presence of alleles encoding weak D, partial D and variant RHCE. Sequence read depth of coverage calculation accurately determined RHD zygosity and detected the presence of RHD/RHCE hybrids. RHCE*C was determined by sequence read depth of coverage and by split read methods. RHD hybrid alleles and RHCE*C were confirmed by using a paired read approach. Small SVs present in RHCE*CeRN and RHCE*ceHAR were detected by a combined read depth of coverage and paired read approach. The combination of several different interpretive approaches allowed for automated software based-RH genotyping of WGS data including RHD zygosity and complex compound RHD and RHCE heterozygotes. The scalable nature of this automated analysis will enable RH genotyping in large genomic sequencing projects.


The effect of ankle taping on detection of inversion-eversion movements in participants with recurrent ankle sprain.
Taping is often used to counter the proprioceptive deficit after joint injury such as ankle sprain. However, the effect of taping on proprioceptive acuity at the ankle is unclear, with conflicting findings. Application of tape improves detection of inversion and eversion movements at the ankle. Controlled laboratory study. The 70% threshold for movement detection was measured in 16 participants with recurrent ankle sprain under 2 conditions: with the ankle taped or untaped. The threshold for movement detection was examined at 3 velocities (0.1 deg/s, 0.5 deg/s, and 2.5 deg/s) and in 2 directions (inversion and eversion). Application of tape significantly decreased the ability to detect movements at the ankle (P < .023). For example, at 0.5 deg/s, the 70% detection threshold was 3.40 degrees +/- 1.05 degrees in inversion and 3.49 degrees +/- 1.15 degrees in eversion at the untaped ankle, and 4.02 degrees +/- 0.86 degrees in inversion and 4.04 degrees +/- 0.89 degrees in eversion at the taped ankle. Taping the ankle decreased the ability to detect movement in the inversion-eversion plane in participants with recurrent ankle sprain. The findings suggest that the efficacy of taping is unlikely to be explained by an enhanced ability to detect inversion or eversion movements. However, because it has been found effective in reducing the incidence of ankle sprain, clinicians should continue taping to reduce the likelihood of resprain.


Using the structure of social networks to map inter-agency relationships in public health services.
This article investigates network governance in the context of health and wellbeing services in England, focussing on relationships between managers in a range of services. There are three aims, namely to investigate, (i) the configurations of networks, (ii) the stability of network relationships over time and, (iii) the balance between formal and informal ties that underpin inter-agency relationships. Latent position cluster network models were used to characterise relationships. Managers were asked two questions, both designed to characterise informal relationships. The resulting networks differed substantially from one another in membership. Managers described networks of relationships that spanned organisational boundaries, and that changed substantially over time. The findings suggest that inter-agency co-ordination depends more on informal than on formal relationships.


Hypoxaemia during open-airway apnoea: a computational modelling analysis.
Hypoxaemia during open-airway apnoea, e.g. during brainstem death testing, may cause organ damage. The effect of ambient oxygen fraction on the extent of hypoxaemia has not been established. We validated the Nottingham Physiology Simulator in this context by reproducing the methodologies and results of four published clinical studies. We then used the simulator to examine the effects of different ambient oxygen fractions (0.21-1.0) and shunt fractions (1-30% of cardiac output) during apnoea. Increasing ambient oxygen fraction from 0.9 to 1.0 more than doubled the time to haemoglobin desaturation at all shunt fractions, and extended apnoea longer than when the ambient oxygen fraction was increased from 0.21 to 0.9. When ambient oxygen fraction and shunt fraction were large, arterial oxygen tension transiently increased during apnoea. A very high ambient oxygen fraction and a patent airway are likely to delay dangerous hypoxaemia during apnoea.


Changes of protein glycosylation in the course of radiotherapy.
This is the first study of changes in protein glycosylation due to exposure of human subjects to ionizing radiation. Site specific glycosylation patterns of 7 major plasma proteins were analyzed; 171 glycoforms were identified; and the abundance of 99 of these was followed in the course of cancer radiotherapy in 10 individual patients. It was found that glycosylation of plasma proteins does change in response to partial body irradiation (∼ 60 Gy), and the effects last during follow-up; the abundance of some glycoforms changed more than twofold. Both the degree of changes and their time-evolution showed large inter-individual variability.


A Gq-Ca2+ axis controls circuit-level encoding of circadian time in the suprachiasmatic nucleus.
The role of intracellular transcriptional/post-translational feedback loops (TTFL) within the circadian pacemaker of the suprachiasmatic nucleus (SCN) is well established. In contrast, contributions from G-coupled pathways and cytosolic rhythms to the intercellular control of SCN pacemaking are poorly understood. We therefore combined viral transduction of SCN slices with fluorescence/bioluminescence imaging to visualize GCaMP3-reported circadian oscillations of intracellular calcium [Ca2+]i alongside activation of Ca2+ /cAMP-responsive elements. We phase-mapped them to the TTFL, in time and SCN space, and demonstrated their dependence upon G-coupled vasoactive intestinal peptide (VIP) signaling. Pharmacogenetic manipulation revealed the individual contributions of Gq, Gs, and Gi to cytosolic and TTFL circadian rhythms. Importantly, activation of Gq-dependent (but not Gs or Gi) pathways in a minority of neurons reprogrammed [Ca2+]i and TTFL rhythms across the entire SCN. This reprogramming was mediated by intrinsic VIPergic signaling, thus revealing a Gq/[Ca2+]i-VIP leitmotif and unanticipated plasticity within network encoding of SCN circadian time.


Taxonomic revision of phlebotomine sand fly species in the series davisi and panamensis of the subgenus Psychodopygus Mangabeira, 1941 (Diptera: Psychodidae: Phlebotominae).
Several species of the subgenus Psychodopygus Mangabeira, 1941 are known to be leishmaniosis vectors in Brazil. Some of them are morphologically similar, which makes their identification quite difficult concerning epidemiological studies. The aim of the current work is to study the morphology of adult specimens of the subgenus Psychodopygus, in accordance with the morphological similarity and still taking into account the epidemiological importance of some species. Thus 11 species have been studied, including four subspecies of adult specimens deposited in the phlebotomine collection of Centro de Pesquisas René Rachou-Fiocruz. Morphological characters found in the literature and new features observed in this study were recorded in a taxonomic discussion format. These characters make it easy to separate such species. Four taxa, previously considered as subspecies, were raised to the category of species.


Initial numbers, serovars and phagevars of Listeria monocytogenes isolated in prepared foods in the city of Barcelona (Spain).
A total of 1100 samples of prepared foods purchased in restaurants and delicatessen shops in the city of Barcelona was examined for the presence of Listerio. L. monocytogenes was more frequently isolated from foods intended to be consumed without further cooking (9.3%) than from foods intended to receive further cooking prior to consumption (2.9%). A quantitative study, carried out with 773 samples, yielded 1% of samples with numbers of L. monocytogenes higher than 100 CFU/g. Strains of L. monocytogenes belonged to serovar 4b (36.3%), 1/2a (29.5%), 1/2b (22.7%) and 1/2c (11.3%). L. monocytogenes serovar 4b phagevar 3274: 2671 (6 of 11) was prevalent among the strains studied.


Novel chitosan/agarose/hydroxyapatite nanocomposite scaffold for bone tissue engineering applications: comprehensive evaluation of biocompatibility and osteoinductivity with the use of osteoblasts and mesenchymal stem cells.
Nanocomposites produced by reinforcement of polysaccharide matrix with nanoparticles are widely used in engineering of biomaterials. However, clinical applications of developed novel biomaterials are often limited due to their poor biocompatibility. The aim of this work was to comprehensively assess biocompatibility of highly macroporous chitosan/agarose/nanohydroxyapatite bone scaffolds produced by a novel method combining freeze-drying with a foaming agent. Within these studies, blood plasma protein adsorption, osteoblast (MC3T3-E1 Subclone 4 and hFOB 1.19) adhesion and proliferation, and osteogenic differentiation of mesenchymal stem cells derived from bone marrow and adipose tissue were determined. The obtained results were also correlated with materials' surface chemistry and wettability to explain the observed protein and cellular response. Obtained results clearly showed that the developed nanocomposite scaffolds were characterized by high biocompatibility and osteoconductivity. Importantly, the scaffolds also revealed osteoinductive properties since they have the ability to induce osteogenic differentiation (Runx2 synthesis) in undifferentiated mesenchymal stem cells. The surface of biomaterials is extremely hydrophilic, prone to protein adsorption with the highest affinity toward fibronectin binding, which allows for good osteoblast adhesion, spreading, and proliferation. Produced by a novel method, macroporous nanocomposite biomaterials have great potential to be used in regenerative medicine for acceleration of the bone healing process.


Influence of age on the effectiveness and duration of protection of Vaxzevria and CoronaVac vaccines: A population-based study.
Aging influences COVID-19 severity and response to vaccination, but previous vaccine effectiveness (VE) analyzes lack the power to evaluate its role in subgroups within the elderly age group. Here we analyzed the impact of age on viral vector and inactivated virus vaccines' effectiveness, the main platforms used in low- and middle-income countries. We report a retrospective longitudinal study of 75,919,840 Brazilian vaccinees from January 18 to July 24, 2021, evaluating documented infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), COVID-19-related hospitalisation, ICU admission, and death. Negative binomial regression models adjusted for sociodemographic characteristics were used for VE estimation. The overall analyzes of full vaccination showed VE against hospitalisation, ICU admission, and death of 91·4% (95%CI:90·1-92·5), 91·1% (95%CI:88·9-92·9) and 92·3% (95%CI:90·5-93·7) for Vaxzevria and 71·2% (95%CI:70·0-72·4), 72·2% (95%CI:70·2-74·0) and 73·7% (95%CI:72·1-75·2) for CoronaVac, respectively. VE for all outcomes is progressively lower with age. In fully-Vaxzevria-vaccinated individuals aged <60 years, VE against death was 96.5% (95%CI:82.1-99.3) versus 68·5% (95%CI:40·0-83·4) in those ≥90 years. Among fully-CoronaVac-vaccinated individuals, VE against death was 84.8% (95%CI:77.1-89.9) in those <60 years compared to 63.5 (95%CI 58.7-67.7) for vaccinees aged 80-89 years and 48·6%; (95%CI:35·0-59·3) for individuals aged ≥90 years. Post-vaccination daily cumulative incidence curves for all outcomes showed increased risk from younger to elder decades of life. There was no increase in the incidence of hospitalisation for individuals <60 years vaccinated during the same period as those aged ≥90 years. Although both vaccines have been effective in protecting against infection, hospitalization and death; Vaxzevria and CoronaVac demonstrated high effectiveness against severe outcomes for individuals up to 79 years of age. Our results reinforce the idea that booster doses should be carefully considered in elders. This study was partially supported by a donation from the "Fazer o bem faz bem" program.


Impact of maternal vaccination timing and influenza virus circulation on birth outcomes in rural Nepal.
To describe the effect of maternal vaccination on birth outcomes in rural Nepal, modified by timing of vaccination in pregnancy and influenza virus activity. A secondary analysis was conducted using data from two annual cohorts of a randomized controlled trial. A total of 3693 pregnant women from Sarlahi District were enrolled between April 25, 2011, and September 9, 2013. All participants were aged 15-40 years and received a trivalent inactivated influenza vaccine or placebo. The outcome measures included birth weight, pregnancy length, low birth weight (<2500 g), preterm birth, and small-for-gestational-age birth. Data were available on birth weight for 2741 births and on pregnancy length for 3623 births. Maternal vaccination increased mean birthweight by 42 g (95% confidence interval [CI] 8-76). The magnitude of this increase varied by season but was greatest among pregnancies with high influenza virus circulation during the third trimester. Birth weight increased by 111 g (95% CI -51 to 273) when 75%-100% of a pregnancy's third trimester had high influenza virus circulation versus 38 g (95% CI -6 to 81) when 0%-25% of a pregnancy's third trimester had high influenza virus circulation. However, these results were nonsignificant. Seasonal maternal influenza vaccination in rural Nepal increased birth weight; the magnitude appeared larger during periods of high influenza virus circulation. CLINICALTRIALS.GOV: NCT01034254.


UTRAD: Anomaly detection and localization with U-Transformer.
Anomaly detection is an active research field in industrial defect detection and medical disease detection. However, previous anomaly detection works suffer from unstable training, or non-universal criteria of evaluating feature distribution. In this paper, we introduce UTRAD, a U-TRansformer based Anomaly Detection framework. Deep pre-trained features are regarded as dispersed word tokens, and represented with transformer-based autoencoders. With reconstruction on more informative feature distribution instead of raw images, we achieve a more stable training process and a more precise anomaly detection and localization result. In addition, our proposed UTRAD has a multi-scale pyramidal hierarchy with skip connections that help detect both multi-scale structural and non-structural anomalies. As attention layers are decomposed to multi-level patches, UTRAD significantly reduces the computational cost and memory usage compared with the vanilla transformer. Experiments on industrial dataset MVtec AD and medical datasets Retinal-OCT, Brain-MRI, Head-CT have been conducted. Our proposed UTRAD out-performs all other state-of-the-art methods in the above datasets. Code released at https://github.com/gordon-chenmo/UTRAD.


TBI Rehabilomics Research: Conceptualizing a humoral triad for designing effective rehabilitation interventions.
Most areas of medicine use biomarkers in some capacity to aid in understanding how personal biology informs clinical care. This article draws upon the Rehabilomics research model as a translational framework for programs of precision rehabilitation and intervention research focused on linking personal biology to treatment response using biopsychosocial constructs that broadly represent function and that can be applied to many clinical populations with disability. The summary applies the Rehabilomics research framework to the population with traumatic brain injury (TBI) and emphasizes a broad vision for biomarker inclusion, beyond typical brain-derived biomarkers, to capture and/or reflect important neurological and non-neurological pathology associated with TBI as a chronic condition. Humoral signaling molecules are explored as important signaling and regulatory drivers of these chronic conditions and their impact on function. Importantly, secondary injury cascades involved in the humoral triad are influenced by the systemic response to TBI and the development of non-neurological organ dysfunction (NNOD). Biomarkers have been successfully leveraged in other medical fields to inform pre-randomization patient selection for clinical trials, however, this practice largely has not been utilized in TBI research. As such, the applicability of the Rehabilomics research model to contemporary clinical trials and comparative effectiveness research designs for neurological and rehabilitation populations is emphasized. Potential points of intervention to modify inflammation, hormonal, or neurotrophic support through rehabilitation interventions are discussed. This article is part of the Special Issue entitled "Novel Treatments for Traumatic Brain Injury".


The hemodynamic response of the conscious pregnant guinea pig to nicotine.
The hemodynamic response to continuous intravenous infusion of nicotine, at 4.5 and 18 micrograms/min, was measured using radiolabeled microspheres in nine chronically catheterized pregnant guinea pigs. These gave serum nicotine levels of 72.3 +/- 6.6 ng/ml and 315 +/- 32 ng/ml (mean +/- SEM), respectively. During low-dose nicotine infusion there was no significant change in cardiac output, its distribution, or uteroplacental blood flow. During high-dose nicotine infusion, cardiac output fell from 257.8 +/- 30.9 ml/min to 212.7 +/- 19.3 ml/min (p less than 0.05) and uteroplacental blood flow fell from 31.2 +/- 3.1 ml/min to 22.3 +/- 2.4 ml/min (p less than 0.05). During control, low-dose, and high-dose periods, serum epinephrine levels rose from control value of 60.2 +/- 2.6 to 98.9 +/- 35 and 1200 +/- 295 pg/ml (p less than 0.05 low dose versus high dose) and serum norepinephrine levels did not change significantly during nicotine infusion. Hence at nicotine levels 20 times but not at two to five times those seen in smokers, modest reductions in cardiac output and uteroplacental blood flow were observed.


Clinical outcomes of pediatric patients with acute abdominal pain and incidental findings of free intraperitoneal fluid on diagnostic imaging.
The presence of free intraperitoneal fluid on diagnostic imaging (sonography or computed tomography [CT]) may indicate an acute inflammatory process in children with abdominal pain in a nontraumatic setting. Although clinical outcomes of pediatric trauma patients with free fluid on diagnostic examinations without evidence of solid-organ injury have been studied, similar studies in the absence of trauma are rare. Our objective was to study clinical outcomes of children with acute abdominal pain of nontraumatic etiology and free intraperitoneal fluid on diagnostic imaging (abdominal/pelvic sonography, CT, or both). We conducted a retrospective review of medical records of children aged 0 to 18 years presenting to a pediatric emergency department with acute abdominal pain (nontraumatic) between April 2008 and March 2009. Patients with intraperitoneal free fluid on imaging were divided into 2 groups: group I, imaging suggestive of an intra-abdominal surgical condition such as appendicitis; and group II, no evidence of an acute surgical condition on imaging, including patients with equivocal studies. Computed tomograms and sonograms were reviewed by a board-certified radiologist, and the free fluid volume was quantitated. Of 1613 patients who underwent diagnostic imaging, 407 were eligible for the study; 134 (33%) had free fluid detected on diagnostic imaging. In patients with both sonography and CT, there was a significant correlation in the free fluid volume (r = 0.79; P < .0005). A significantly greater number of male patients with free fluid had a surgical condition identified on imaging (57.4% versus 25%; P < .001). Children with free fluid and an associated condition on imaging were more likely to have surgery (94.4% versus 6.3%; P < .001). We found clinical outcomes (surgical versus nonsurgical) to be most correlated with a surgical diagnosis on diagnostic imaging and not with the amount of fluid present.


Rapid cloning of antigen-specific T-cell receptors by leveraging the cis activation of T cells.
It is commonly understood that T cells are activated via trans interactions between antigen-specific T-cell receptors (TCRs) and antigenic peptides presented on major histocompatibility complex (MHC) molecules on antigen-presenting cells. By analysing a large number of T cells at the single-cell level on a microwell array, we show that T-cell activation can occur via cis interactions (where TCRs on the T cell interact with the antigenic peptides presented on MHC class-I molecules on the same cell), and that such cis activation can be used to detect antigen-specific T cells and clone their TCR within 4 d. We used the detection-and-cloning system to clone a tumour-antigen-specific TCR from peripheral blood mononuclear cells of healthy donors. TCR cloning by leveraging the cis activation of T cells may facilitate the development of TCR-engineered T cells for cancer therapy.


Volume-sensitive taurine transport in fish erythrocytes.
Taurine plays an important role in cell volume regulation in both vertebrates and invertebrates. Erythrocytes from two euryhaline fish species, the eel (Anguilla japonica) and the starry flounder (Platichthys stellatus) were found to contain high intracellular concentrations of this amino acid (approximately equal to 30 mmol per liter of cell water). Kinetic studies established that the cells possessed a saturable high-affinity Na+-dependent beta-amino-acid transport system which also required Cl- for activity (apparent Km (taurine) 75 and 80 microM; Vmax 0.85 and 0.29 mumol/g Hb per hr for eel (20 degrees C) and flounder cells (10 degrees C), respectively. This beta-system operated with an apparent Na+/Cl-/taurine coupling ratio of 2:1:1. A reduction in extracellular osmolarity, leading to an increase in cell volume, reversibly decreased the activity of the transporter. In contrast, low medium osmolarity stimulated the activity of a Na+-independent nonsaturable transport route selective for taurine, gamma-amino-n-butyric acid and small neutral amino acids, producing a net efflux of taurine from the cells. Neither component of taurine transport was detected in human erythrocytes. It is suggested that these functionally distinct transport routes participate in the osmotic regulation of intracellular taurine levels and hence contribute to the homeostatic regulation of cell volume. Volume-induced increases in Na+-independent taurine transport activity were suppressed by noradrenaline and 8-bromoadenosine-3', 5'-cyclic monophosphate, but unaffected by the anticalmodulin drug, pimozide.


Histotopography and ultrastructure of the thin limbs of the loop of Henle in the hamster.
In the kidney of the Syrian hamster the descending thin limbs of both the short and long loops of Henle are not spatially separated from each other and descend between the vascular bundles. Ultrastructurally, five different epithelial types are distinguished in the thin limbs of the short and long loops of Henle. Short loops possess only a descending thin limb with a simply organized epithelium (type 1). Long loops comprise an upper and a lower part of the descending thin limb and the ascending thin limb. The upper part of the long descending thin limb is equipped with a complex and highly interdigitating epithelium with shallow junctions (type 2), which gradually transforms into the simple noninterdigitating type-3 epithelium of the lower part. In a minor portion of long descending thin limbs, however, the upper part begins with an even more complexly organized epithelium (type 2a) than type 2. Type-2a epithelium is conspicuously thicker and possesses a more elaborate mode of cellular interdigitation. Along the descent of this tubular part through the inner stripe of the outer medulla, type-2a epithelium transforms into type-2 epithelium. It is suggested that the long descending thin limbs, which start with type-2a epithelium, belong to the longest loops. The type-4 epithelium of the ascending thin limbs is characterized by flat and extensively interdigitating cells with shallow junctions. The unique pattern of the type-2a epithelium favors the assumption that solute secretion essentially contributes to the increase in concentration of tubular fluid in long descending thin limbs.


Efficiency and cost effectiveness: PAGE-SSCP versus MDE and Phast gels for the identification of unknown beta thalassaemia mutations.
Prenatal diagnosis for beta thalassaemia has proved to be very effective in preventing the birth of an affected child and hence in controlling the disease. The success of prenatal diagnosis depends on the delineation of the underlying mutations in the population at risk. Each population carries a limited number of frequent defects (89-91%) and a variable number of rare alleles (4-5%), whereas 2-3% of alleles remain uncharacterised. To offer prenatal diagnosis when the parental mutation is unknown, the application of a non-specific detection method (such as single stranded conformational polymorphism (SSCP)) to localise the mutation, followed by direct sequencing of the amplified gene sequence, is required. With this objective in mind, this study was designed to devise the best protocol and system of SSCP for the rapid screening of unknown mutations in the beta globin gene. To detect mutations in this disease, three different systems-Phast gels, MDE gels, and polyacrylamide gels-were used under varying conditions. Polyacrylamide gels were found to be the most efficient, both in terms of resolution and cost. Polyacrylamide gels are the most rapid, efficient, reliable, and cost effective means for DNA mutation analysis of the beta globin gene.


Psychological distress after initial treatment for breast cancer: a comparison of partial and total mastectomy.
This study of the relation between type of mastectomy and psychological distress was based on all newly diagnosed breast cancer patients with localized or regional disease who were surgically treated in seven Quebec City hospitals in 1984. Among 235 eligible patients, 227 (96%) participated in a home interview 3 months after diagnosis and 205 of these women gave a second interview approximately 15 months later. The Psychiatric Symptom Index (PSI) was used to measure the severity of symptoms of psychological distress. At 3 months, 38.8% of partial mastectomy patients had high PSI scores compared to 25.8% of women treated by total mastectomy (OR = 1.8, p = 0.044). Fifteen months later, percentages with high scores were identical, 35.1%, in the two treatment groups. Age appeared to modify the surgery-distress relation. These results suggest that partial mastectomy does not protect against psychological distress after breast cancer. Moreover, they highlight the importance of adequate preparation and support for all breast cancer patients, regardless of type of initial surgery.


Synthesis and turnover of the putative T-lymphocyte antigen receptor.
Cells of an antigen-specific T-cell clone, A37.4, were treated with the proteolytic enzymes trypsin or pronase to remove the T-cell antigen receptor. Removal of the receptor, analysed by surface labelling and non-reduced/reduced gel electrophoresis, stimulated the cells to synthesize new protein rapidly. New membrane protein was readily detectable within 3 h after reculture, and it was able to interact with antigen-presenting cells, since the T cells could be stimulated to produce interferon. Treatment of cells with metabolic inhibitors demonstrated that there was only a small cytoplasmic pool of protein, and de novo synthesis of mRNA was necessary for quantitative replacement of the membrane protein. The protein also required glycosylation for transport and insertion into the membrane. Despite rapid resynthesis of new receptor, when surface-labelled cells were recultured either with or without antigen-presenting cells, there was not a rapid turnover of the receptor. There was also no evidence of any loss of receptor from the membrane of antigen-stimulated cells or of any shedding of receptor into the culture medium.


Photodiagnosis and photodynamic therapy of peritoneal metastasis of ovarian cancer.
Ovarian cancer is among the deadliest in women. Current treatment strategies fail to cure many patients owing to the difficulties of eradicating peritoneal implants frequently associated with this pathology. Photodynamic therapy represents a promising treatment as it offers many advantages over alternative strategies: diagnostic properties, specific targeting of abnormal cells, possibility to be combined with other therapies.


Obesity-related cardiometabolic indicators modify the associations of personal noise exposure with heart rate variability: A further investigation on the Study among Obese and Normal-weight Adults (SONA).
Elucidating the associations between environmental noise and heart rate variability (HRV) would be beneficial for the prevention and control of detrimental cardiovascular changes. Obese people have been found to manifest heightened susceptibility to the adverse effects of noise on HRV. However, the underlying mechanisms remain unclear. Based on 53 normal-weight and 44 obese young adults aged 18-26 years in Beijing, China, this study aimed to investigate the role of obesity-related cardiometabolic indicators for associations between short-term environmental noise exposure and HRV in the real-world context. The participants underwent personal noise exposure and ambulatory electrocardiogram monitoring using portable devices at 5-min intervals for 24 continuous hours. Obesity-related blood pressure, glucose and lipid metabolism, and inflammatory indicators were subsequently examined. Generalized mixed-effect models were used to estimate the associations between noise exposure and HRV parameters. The C-peptide, homeostasis model assessment of insulin resistance (HOMA-IR), and leptin levels were higher in obese participants compared to normal-weight participants. We observed amplified associations between short-term noise exposure and decreases in HRV among participants with higher C-peptide, HOMA-IR, and leptin levels. For instance, a 1 dB(A) increment in 3 h-average noise exposure level preceding each measurement was associated with changes of -0.20% (95%CI: -0.45%, 0.04%) and -1.35% (95%CI: -1.85%, -0.86%) in standard deviation of all normal to normal intervals (SDNN) among participants with lower and higher C-peptide levels, respectively (P for interaction <0.05). Meanwhile, co-existing fine particulate matter (PM2.5) could amplify the associations between noise and HRV among obese participants and participants with higher C-peptide, HOMA-IR, and leptin levels. The more apparent associations of short-term exposure to environmental noise with HRV and the effect modification by PM2.5 may be partially explained by the higher C-peptide, HOMA-IR, and leptin levels of obese people.


Isolation of a polymorphic CA repeat sequence at the human progesterone receptor (PGR) locus.
We isolated a polymorphic dinucleotide (CA) repeat sequence from a genomic clone containing the human progesterone receptor (PGR) gene. This polymorphism will be a useful marker in the genetic study of disorders affecting female endocrine systems, such as progesterone resistance and breast, uterine, and ovarian cancers.


Collaboratives grow, even among traditional health care competitors.
The IHI-pioneered concept already is generating positive results. Participants gain access to panels of experts and evidence-based best practices. Challenges seen in sustaining change once collaborative has been disbanded.


Systems identification and characterization of β-glucuronosyltransferase genes involved in arabinogalactan-protein biosynthesis in plant genomes.
Utilizing plant biomass for bioethanol production requires an understanding of the molecular mechanisms involved in plant cell wall assembly. Arabinogalactan-proteins (AGPs) are glycoproteins that interact with other cell wall polymers to influence plant growth and developmental processes. Glucuronic acid, which is transferred to the AGP glycan by β-glucuronosyltransferases (GLCATs), is the only acidic sugar in AGPs with the ability to bind calcium. We carried out a comprehensive genome-wide analysis of a putative GLCAT gene family involved in AGP biosynthesis by examining its sequence diversity, genetic architecture, phylogenetic and motif characteristics, selection pressure and gene expression in plants. We report the identification of 161 putative GLCAT genes distributed across 14 plant genomes and a widely conserved GLCAT catalytic domain. We discovered a phylogenetic clade shared between bryophytes and higher land plants of monocot grass and dicot lineages and identified positively selected sites that do not result in functional divergence of GLCATs. RNA-seq and microarray data analyses of the putative GLCAT genes revealed gene expression signatures that likely influence the assembly of plant cell wall polymers which is critical to the overall growth and development of edible and bioenergy crops.


Temperature- and pH-dependent effect of lactate on in vitro redox stability of red meat myoglobins.
Our objective was to evaluate the influence of lactate on in vitro redox stability and thermostability of beef, horse, pork, and sheep myoglobins. Lactate (200 mM) had no effect (P>0.05) on redox stability at physiological (pH7.4, 37°C) and meat (pH 5.6, 4°C) conditions. However, lactate increased (P<0.05) metmyoglobin formation at a condition simulating stressed live skeletal muscle (pH 6.5, 37°C). The redox stability of myoglobins at stressed live skeletal muscle and meat conditions was species-specific (P<0.05). Myoglobin thermostability at 71°C was lower (P<0.05) in the presence of lactate compared with controls and was influenced (P<0.05) by species. The results of the present study indicate that the effects of lactate on myoglobin are temperature and pH dependent. The observed lack of influence of lactate on myoglobin redox stability at meat condition suggests that the color stability of lactate-enhanced fresh meat is not due to direct interactions between the ingredient and the heme protein.


Green one-step synthesis of carbon quantum dots from orange peel for fluorescent detection of Escherichia coli in milk.
Carbon quantum dots (CQDs) prepared by a green one-step approach was used for sensitive and selective assay of Escherichia coli O157: H7 (E. coli). CQDs was synthesized from orange peel as a carbon source via a microwave-assisted method. The CQDs displayed strong green fluorescence under excitation wavelength of 420 nm. A fluorescent probe (CQDs-MNPs) for E. coli was fabricated based on CQDs labeled with aptamer (aptamer-CQDs) and magnetic nanoparticles labeled with complementary DNA (cDNA-MNPs). Fluorescent intensity of the CQDs-MNPs was decreased with addition of E. coli. The linearity between fluorescent intensity and E. coli concentration was used for developing a fluorescent method with detecting range of 500-106 CFU/mL and detection limit of 487 CFU/mL. Milk samples contaminated by E. coli were analyzed by this method, and the results agreed with that achieved by plate-counting methods. This fluorescent probe exhibits great potential in guaranteeing food quality and safety.


Pivampicillin-promoted excretion of pivaloylcarnitine in humans.
Pivampicillin treatment of seven children (five boys and two girls) for 7 days significantly reduced the amounts of total acid-soluble carnitine, free carnitine, and long-chain acylcarnitines and increased the amounts of acid-soluble acylcarnitine in plasma. The fasting plasma levels of 3-hydroxybutyrate at the end of treatment were 15% of the control value. The levels of free fatty acids were decreased, whereas triglyceride levels were unaffected, indicating impaired fat metabolism. Daily urinary excretion of total carnitine was four to five times higher than controls after the first day of treatment, although the amounts of free carnitine and acetylcarnitine were decreased. The urinary acylcarnitines were isolated and characterized by gas chromatography/electron impact mass spectrometry and fast-atom bombardment mass spectrometry. Pivaloylcarnitine was the predominant urinary acylcarnitine; it represented greater than 96% of the increased excretion of total carnitine and 75-80% of the total conjugated pivalic acid. The renal clearance of acylcarnitines was comparable to that of creatinine, indicating no reabsorption of pivaloylcarnitine. These data suggest a detoxification function of carnitine for pivalic acid in humans.


Posttraumatic coronary artery aneurysm.
A 31-year-old man sustained blunt deceleration trauma with dissection of the left anterior descending coronary artery. We repaired a posttraumatic coronary aneurysm to prevent late occlusion of the recanalized vessel. An internal mammary artery graft was applied as a precaution against postoperative thrombosis. Follow-up at 12 months showed improved left ventricular function.


Exploring the Relationship of a Gamified Assessment with Performance.
Our study explores the validity of a game-based assessment method assessing candidates' soft skills. Using self-reported measures of performance, (job performance, Organizational Citizenship Behaviors (OCBs), and Great Point Average (GPA), we examined the criterion-related and incremental validity of a game-based assessment, above and beyond the effect of cognitive ability and personality. Our findings indicate that a game-based assessment measuring soft skills (adaptability, flexibility, resilience and decision making) can predict self-reported job and academic performance. Moreover, a game-based assessment can predict academic performance above and beyond personality and cognitive ability tests. The effectiveness of gamification in personnel selection is discussed along with research and practical implications introducing recruiters and HR professionals to an innovative selection technique.


Inhibition of renal dipeptidyl peptidase IV enhances peptide YY1-36-induced potentiation of angiotensin II-mediated renal vasoconstriction in spontaneously hypertensive rats.
Dipeptidyl peptidase IV inhibitors are a new class of antidiabetic drugs. It is urgent, therefore, to fully understand the pharmacology of these inhibitors. Although dipeptidyl peptidase IV metabolizes at least 24 endogenous substrates, the pharmacological consequences of inhibiting the metabolism of most of these substrates is unknown. Our previous results show that Y(1) receptors, but not Y(2) receptors, enhance renovascular responses to angiotensin II in kidneys from genetically susceptible animals (spontaneously hypertensive rats). Dipeptidyl peptidase IV converts peptide YY(1-36) (circulating hormone) to peptide YY(3-36), and peptide YY(1-36) is a Y(1)-receptor agonist, whereas peptide YY(3-36) is a selective Y(2)-receptor agonist. Therefore, it is conceivable that inhibition of dipeptidyl peptidase IV in genetically susceptible kidneys may increase the ability of peptide YY(1-36) to potentiate angiotensin II-induced renal vasoconstriction. Here we demonstrate that in kidneys from spontaneously hypertensive rats 1) peptide YY(1-36) potentiates renovascular responses to angiotensin II, whereas peptide YY(3-36) has little effect, 2) 3-N-[(2S,3S)-2-amino-3-methylpentanoyl]-1,3-thiazolidine (P32/98) (dipeptidyl peptidase IV inhibitor) augments the ability of peptide YY(1-36) to enhance renovascular responses to angiotensin II, 3) dipeptidyl peptidase IV is expressed in preglomerular microvessels and glomeruli, 4) kidneys metabolize arterial PYY(1-36) to PYY(3-36) via a mechanism blocked by P32/98, and 5) preglomerular microvessels and glomeruli convert peptide YY(1-36) to peptide YY(3-36), and this conversion is inhibited by P32/98. We conclude that dipeptidyl peptidase IV is expressed in the renal microcirculation and inhibition of this ecto-enzyme causes arterial PYY(1-36) to more effectively enhance angiotensin II-induced renal vasoconstriction in genetically susceptible kidneys.


Hypoxia stress affects the physiological responses, apoptosis and innate immunity of Kuruma shrimp, Marsupenaeus japonicus.
For commercial aquatic animals, hypoxia phenomenon often occurs in live transport and aquaculture. In previous studies, much interest has been focused on antioxidant enzyme activities and could not present the complexities. The multifaceted responses, especially considering physiological indexes, histological structure, cell apoptosis, and immune pathways, are still unknown. In this study, we investigated the comprehensive hypoxic responses of Marsupenaeus japonicus. The results showed that the physiological indexes showed time-dependent changes upon hypoxia stress. Hypoxia stress led to significant tissue damage and cell apoptosis in the gill and hepatopancreas. Compared with the control group, the apoptosis index (AI) of the 12 h hypoxic treatment increased significantly (p < 0.05) in the gills and hepatopancreas. Comparative transcriptome analysis identified 900 and 1400 differentially expressed genes (DEGs) in the gill and hepatopancreas, respectively. Several DEGs were related to the lysosome, glycolysis/gluconeogenesis, citrate cycle, and apoptosis, and seven of them were validated using quantitative real-time PCR. This study provided valuable clues to understanding the mechanisms underlying the hypoxic responses of M. japonicus.


The mechanism of inhibition of hepatitis B virus replication by the carbocyclic analog of 2'-deoxyguanosine.
The carbocyclic analog of deoxyguanosine inhibits hepatitis B virus replication by greater than 95% in the hepatitis B virus-producing cell line (2.2.15) as monitored by decreases of secreted hepatitis B virus DNA, hepatitis B virus polymerase activity and intracellular episomal hepatitis B virus DNA. Transcription of hepatitis B virus RNA from chromosomally integrated hepatitis B virus DNA was unaffected. Radioactive carbocyclic 2'-deoxyguanosine was directly phosphorylated within the 2.2.15 cells and was incorporated exclusively into DNA. In contrast, radioactive deoxyguanosine was presumably metabolized through the "salvage" pathway in which the guanine was primarily incorporated into cellular RNAs. The rate of incorporation of carbocyclic 2'-deoxyguanosine in 2.2.15 cells was similar to that in the parental cell line (HepG2), which does not contain hepatitis B virus sequences. Greater than 90% of the analog was present at internal sites within the DNA, indicating that the analog did not function as a DNA chain terminator. Kinetic analysis of the Km and Ki of dGTP and carbocyclic 2'-deoxyguanosine 5'-triphosphate, respectively, using both hepatitis B virus polymerase and DNA polymerase delta indicated that the analog is a competitive inhibitor for dGTP. Although both polymerases had similar Km's for dGTP, the Ki for carbocyclic 2'-deoxyguanosine 5'-triphosphate was about 6 times lower using the hepatitis B virus polymerase. This would indicate that, at low concentrations of intracellular carbocyclic 2'-deoxyguanosine 5'-triphosphate, the hepatitis B virus polymerase would be preferentially inhibited. We propose this to be the mechanism acting to inhibit preferentially hepatitis B virus replication in the tissue culture cells.


Effect of low mole fraction of trehalose dicorynomycolate from Corynebacterium diphtheriae on water permeability and electrical capacitance of lipid bilayer membranes.
The effect of incorporation of different proportions of trehalose dicorynomycolates (TDC) into lecithin bilayer membranes was studied. It was found that TDC, induces a 14% decrease of water osmotic permeability (42.6 +/- 3.9 to 36.8 +/- 2.7 microns/s) at 1.6 mole%, suggesting that this substance leads to an increase of the degree of packing of the constituent lipid molecules. A condensing effect of TDC was also apparent from membrane electrical capacitance (Cm) measurement. By incorporating TDC into bilayer membranes, the value of Cm experienced a decrease of 29% at 1.6% mole fraction. This finding was taken to reflect an increase in membrane thickness, known in many examples, to be related to the condensing effect.


Potential Implications of Angiotensin-converting Enzyme 2 Blockades on Neuroinflammation in SARS-CoV-2 Infection.
Angiotensin-converting enzyme 2 (ACE2) has been reported as a portal for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Consequently, scientific strategies to combat coronavirus disease of 2019 (COVID-19) were targeted to arrest SARS-CoV-2 invasion by blocking ACE2. While blocking ACE2 appears a beneficial approach to treat COVID-19, clinical concerns have been raised primarily due to the various intrinsic roles of ACE2 in neurological functions. Selective reports indicate that angiotensin receptor blockers (ARBs) and angiotensin-converting enzyme inhibitors (ACEIs) upregulate ACE2 levels. ACE2 metabolizes angiotensin II and several peptides, including apelin-13, neurotensin, kinetensin, dynorphin, (des-Arg9) bradykinin, and (Lys-des-Arg9)-bradykinin, which may elicit neuroprotective effects. Since ARBs and ACEIs upregulate ACE2, it may be hypothesized that patients with hypertension receiving ARBs and ACEIs may have higher expression of ACE2 and thus be at a greater risk of severe disease from the SARS-CoV-2 infections. However, recent clinical reports indicate the beneficial role of ARBs/ACEIs in reducing COVID-19 severity. Together, this warrants a further study of the effects of ACE2 blockades in hypertensive patients medicated with ARBs/ACEIs, and their consequential impact on neuronal health. However, the associations between their blockade and any neuroinflammation also warrant further research. This review collates mechanistic insights into the dichotomous roles of ACE2 in SARSCoV- 2 invasion and neurometabolic functions and the possible impact of ACE2 blockade on neuroinflammation. It has been concluded that ACE2 blockade imposes neuroinflammation.


Inhibitory effects of bevacizumab on angiogenesis and corneal neovascularization.
To evaluate the inhibitory effects of bevacizumab (Avastin) on angiogenesis using cultured human umbilical vein endothelial cells (HUVECs) in vitro and on corneal neovascularization by subconjunctival injection of bevacizumab in vivo. After the HUVECs were exposed to different concentrations of bevacizumab stimulated with VEGF (10 ng/ml) for 2, 6, and 24 hours, cellular-activity-like proliferation, migration and tube formation were assessed. Subconjunctival injection of bevacizumab (2.5 mg/0.1 ml) was performed after corneal chemical burn injury. Then the cornea was evaluated by biomicroscopy, fluorescein angiography, and light microscopy. The inhibitory effects of bevacizumab on VEGF-induced HUVECs proliferation showed a dose-dependent response for 2 and 6 hours, but all groups were effectively inhibited regardless of the concentration of bevacizumab for 24 hours. The inhibitory effects of bevacizumab on the migration of VEGF-induced HUVECs showed a time- and dose-dependent response. The inhibitory effects of bevacizumab on VEGF-induced HUVECs tube formation showed a dose-dependent response only for 24 hours. On days 3 and 8 after the subconjunctival injection, bevacizumab-treated eyes showed less neovascular growth than BSS-treated eyes in biomicroscopic, fluorescein angiographic, and light microscopic findings in vivo. Bevacizumab effectively inhibits angiogenesis and corneal neovascularization, and could be used as a inhibitor of corneal neovascularization in the future.


Cushing's disease: towards precision medicine.
The pathogenesis of Cushing's disease is poorly understood; two recent reports identifying somatic mutations in USP8 in pituitary corticotroph tumors provide exciting advances in this field. These mutations alter EGFR trafficking and signaling, raising the prospect that EGFR inhibitors may move the treatment of this disease into the era of precision medicine.


Group mentoring: a story of transition for undergraduate baccalaureate nursing students.
Group mentoring has been endorsed as an effective method of supporting novice professionals across disciplines. In one university, faculty revised the undergraduate nursing curriculum to include a group mentoring course as a requirement of students during the four semesters they are enrolled in the nursing program. The purpose of this study was to explore the lived experience of undergraduate nursing students participating in a group mentoring course. This study used a hermeneutic interpretive phenomenological method. Data were collected from 22 undergraduate nursing students enrolled in group mentoring courses at a private Midwestern university in the United States. At the end of each semester of mentoring, students provided written responses to five open-ended questions about their experiences of participating in the mentoring courses. Four themes emerged: conversation, communication, connection, and cohesion. Group mentoring was an effective way to support nursing students as they transitioned from undergraduate student to novice professional nurse.


Patterns of rescue and maintenance therapy claims surrounding a clinical encounter for an asthma exacerbation.
A "window of opportunity" has been proposed where anti-inflammatory therapy administration in response to symptoms could prevent exacerbation. To evaluate rescue and maintenance therapy claims surrounding a severe asthma exacerbation serious enough to require a face-to-face clinical encounter. Merative MarketScan research databases (US administrative claims 2011 to 2017) were analyzed for patients aged ≥4 years, with an asthma diagnosis code, who filled short-acting β2-agonist (SABA) and Global Initiative for Asthma Steps 3 to 5 maintenance therapies. Patients were indexed on a random SABA claim and had 12 months' continuous health plan eligibility pre- and post-index. Serious exacerbations were severe exacerbations requiring systemic corticosteroids prescribed from an outpatient clinic, urgent care or emergency department, or hospitalization for asthma. SABA and maintenance claims 30 days pre- and post-event were analyzed. Of 319,342 patients (30% children 4 to 11 years; 70% adults or adolescents ≥12 years), 27.2% of children and 16.8% of adolescents or adults experienced ≥ 1 serious exacerbation (unadjusted odds ratio [OR], 1.85 [95% confidence interval, 1.81-1.88]). In the 30 days pre-event, 42.6% filled ≥1 SABA (children: 44.3%; adolescents or adults: 41.5%; OR, 1.12 [1.09-1.16]) and 57.4% filled maintenance (children: 59.0%; adolescents or adults: 56.3%; OR, 1.12 [1.08-1.15]). In the 30 days post-event, 61.4% filled SABA (children: 69.7%; adolescents or adults: 55.6%; OR, 1.84 [1.78-1.90]) and 94.8% filled maintenance (children: 98.6%; adolescents or adults: 92.2%; OR, 6.09 [5.45-6.81]). Many patients treated as having moderate-to-severe asthma escalate SABA claims before a serious exacerbation, but approximately 40% have no anti-inflammatory maintenance fill, highlighting a "window of opportunity" to prevent exacerbations using inhaled corticosteroids concomitantly with SABA as rescue.


Secondary alveolar cleft grafting using autogenous mineralized plasmatic matrix (MPM) versus cancellous bone particles derived from anterior iliac crest.
The essential concern of alveolar cleft grafting in patients of cleft lip and palate at the mixed dentition phase is to gain bone within the cleft area that provides closure of the oronasal communication with continuous and stable maxilla for future cleft teeth eruption or implantation. This study aimed to compare the effectiveness of mineralized plasmatic matrix (MPM) versus cancellous bone particles harvested from anterior iliac crest in secondary alveolar cleft grafting. This prospective randomized controlled trial was conducted on ten patients with unilateral complete alveolar cleft requiring cleft reconstruction. Patients were randomly divided into two equal groups; group (1) included 5 patients who received particulate cancellous bone derived from anterior iliac crest (control group) and group (2) included 5 patients who received MPM graft prepared from cancellous bone derived from anterior iliac crest (study group). All patients received CBCT preoperatively, immediately postoperatively and after 6 months. On the CBCT, graft's volume, labio-palatal width, and height were measured and compared. The outcome of the studied patients 6 months postoperatively showed that the control group had significant decrease in the graft volume, labio-palatal width, and height compared to the study group. MPM allowed for the integration of bone graft particles inside a fibrin network, which offers positional stability of the bone particles, thus preserving their shape with subsequent "in situ" immobilization of the graft components. This conclusion was reflected positively in terms of maintained graft volume, width, and height compared to that of the control group. MPM allowed for maintenance of grafted ridge volume, width, and height.


Ex vivo expansion of hematopoietic stem cells from human cord blood in serum-free conditions.
Human cord blood (CB), collected from the postpartum placenta and umbilical cord, has been identified as a rich source of hematopoietic stem cells (HSCs) and provides an alternative to bone marrow or mobilized peripheral blood transplantation. However, the major restriction of CB transplantation is the low number of HSCs in each CB unit and limits its use in clinical transplantation. The development of ex vivo culture systems that facilitate the expansion of HSCs is crucial to stem cell research and clinical application. In this chapter, we describe the protocols to isolate HSCs from CB, expand HSCs in serum-free condition, and analyze HSCs. This information is benefical for successful use of CB stem cells in therapeutic studies.


Effects of addition of different vegetable oils to lactating dairy ewes' diet on meat quality characteristics of suckling lambs reared on the ewes' milk.
The effect of different vegetable oils used in the diet of lactating ewes on the meat quality of their suckling lambs has been evaluated. Lambs (males and females) were slaughtered at 11 kg. Fortyeight lactating Churra ewes (prolificacy 1.5) and their suckling lambs were assigned to four treatments according to the oil added (3% on weight basis) to the ewes' daily ration: palm oil as control (CON); olive oil (OLI); soybean oil (SOY); and linseed oil (LIN). Analyses of pH, colour, thiobarbituric acid reactive substances (TBARS), tocopherol levels, volatile compounds and a sensory evaluation were carried out on suckling lambs' meat. Results showed no substantial effect on pH, colour, TBARS and tocopherol levels. Volatiles typically derived from lipid oxidation were higher in SOY group. However, panellists were only able to correctly identify samples from LIN group. Furthermore, the meat from LIN group showed lower scores towards odour and flavour quality and overall liking than that from the rest of treatments.


Anatomic Concordance of Neonatologist-Performed Echocardiography as Part of Hemodynamics Consultation and Pediatric Cardiology.
Targeted neonatal echocardiography (TnECHO) performed by neonatologists as part of a hemodynamics consultation is increasingly being used in neonatal intensive care units. To minimize delays in obtaining physiologic data, first echocardiograms may be obtained by the neonatal hemodynamics team and reviewed afterward by a pediatric cardiologist. This practice has not been systematically evaluated. The aim of this study was to compare concordance between anatomic findings on TnECHO and pediatric cardiology reports. This was a retrospective evaluation of 339 infants at low risk for congenital heart disease (CHD) admitted to two large referral centers with established neonatal hemodynamics programs who underwent comprehensive TnECHO as their first postnatal echocardiographic examinations. The protocol included comprehensive imaging of intracardiac anatomy, outflow tract concordance and integrity, aortic arch anatomy, pulmonary vein location and flow, and transitional shunts. The hemodynamics consultation note was compared with the cardiology report to determine anatomic concordance or major or minor discrepancies in all first studies. Anatomic concordance occurred in 97.9% (κ = 0.862; 95% CI, 0.762-0.962; P < .001). There were seven minor discrepancies (small muscular ventricular septal defects and coronary fistulas). The index population included 23 infants (6.7%) with CHD, of whom only one (0.3%) had a ductal-dependent lesion (coarctation of the aorta) which was correctly identified by both teams. The rate of major CHD in patients considered eligible for hemodynamics consultation was low, and there was high diagnostic concordance between trained neonatal hemodynamics specialists and pediatric cardiology. First echocardiograms obtained by subspecialty neonatologists may provide imaging of sufficient quality to evaluate a critically unwell neonate with low suspicion for critical CHD lesions. These results should not be extrapolated to infants in whom CHD is suspected. This study highlights the importance of formalized, rigorous, and standardized training for neonatologists with hemodynamics expertise who perform timely assessments using TnECHO.


Effect of laser therapy on immune cells infiltrate after excisional wounds in diabetic rats.
Diabetes alters innate and specific immunity, causing an imbalanced tissue repair process. Very active neutrophils and macrophages are found for a long time in chronic wounds in those individuals. The aim of this study was to evaluate the response of the main effector cells of immunity (neutrophils, macrophages, and T lymphocytes) and to compare the effects of two laser therapy regimens in the postoperative treatment of excision wounds. Diabetes was induced in female Wistar rats and a punch was used to cause wounds in the dorsum of each individual. The animals were randomly allocated to a control group (CG), in which the wound was untreated, a single-dose laser group (SLG), in which the wound was submitted to single dose of laser therapy at wavelength of 660 nm, output power of 30 mW, energy density of 4 J/cm(2), and 26-second exposure time, and a fractionated-dose laser group (FLG), submitted to 1 J/cm(2) of laser therapy on Days 1, 3, 8, and 10. Euthanasia was performed on five animals from each group Days 1, 3, 8, 10, 15, and 22. The wound was removed and routinely processed for immunohistochemistry against elastase, CD3, CD68, and CD206 antibodies. The samples were photographed and labeled cells were counted by a blinded observer. The Kruskal-Wallis test was used for the statistical analysis. Neutrophils were predominant in the SLG on Day 1, whereas these cells were mostly found in the CG on Day 3 (P < 0.05). The T lymphocyte count was similar in all groups in the throughout the experiment. On Day 3, the SLG exhibited a greater number of total macrophages (CD68+) (P < 0.05), whereas the macrophage count was similar among the different groups on the other evaluation days. The CD206 + cell counts revealed that the SLG had more M2 macrophages than the CG on Day 8 (P < 0.05), whereas the FLG exhibited more M2 macrophages than the CG on Day 10 (P < 0.05). The present findings demonstrate that laser therapy can alter the composition of inflammatory infiltrate in diabetic wounds, leading to a more balanced response transiting from a rapid neutrophil infiltration through to M2 macrophage polarization, especially with a single application of 4 J/cm(2) in the immediate postoperative period.


An unusual case which began with subcutaneous panniculitis followed by fever, severe hepatic involvement and hyperlipidemia.
A 25-year-old man was at first dermatologically suspected as suffering from Weber-Christian syndrome because of subcutaneous panniculitis, but his skin lesions disappeared completely during the course. Hyperlipidemia, disturbances in liver function, and leukemoid reaction became remarkable and he died of subarachnoid hemorrhage eleven months after onset. Necropsy revealed subarachnoid hemorrhage at the base of the brain, lipogranulomatous and inflammatory lesions in the upper lobe of the left lung, a remarkable fatty liver, splenomegaly, pericarditis, and foam cells in the spleen, liver, and bone marrow. A comparison with 57 autopsy cases of Weber-Christian syndrome reported in the literature showed our case to be an exceptional instance of Weber-Christian syndrome, if the present case is not to be regarded as a different disease entity.


Phylogenetic relationships and biogeography of Podarcis species from the Balkan Peninsula, by bayesian and maximum likelihood analyses of mitochondrial DNA sequences.
Wall lizards of the genus Podarcis (Sauria, Lacertidae) comprise 17 currently recognized species in southern Europe, where they are the predominant nonavian reptile group. The taxonomy of Podarcis is complex and unstable. Based on DNA sequence data, the species of Podarcis falls into four main groups that have substantial geographic coherence (Western island group, southwestern group, Italian group, and Balkan Peninsula group). The Balkan Peninsula species are divided into two subgroups: the subgroup of P. taurica (P. taurica, P. milensis, P. gaigeae, and perhaps P. melisellensis), and the subgroup of P. erhardii (P. erhardii and P. peloponnesiaca). In the present study, the question of phylogenetic relationships among the species of Podarcis encountered in the Balkan Peninsula was addressed using partial mtDNA sequences for cytochrome b (cyt b) and 16S rRNA (16S). The data support the monophyly of Podarcis and suggest that there are three phylogenetic clades: the clade A (P. taurica, P. gaigeae, P. milensis, and P. melisellensis); the clade B (P. erhardii and P. peloponnesiaca), and the clade C (P. muralis and P. sicula). By examining intraspecific relationships it was found that extant populations of P. erhardii are paraphyletic. Furthermore, subspecies previously defined on the basis of morphological characteristics do not correspond to different molecular phylogenetic clades, suggesting that their status should be reconsidered. The distinct geographic distribution of the major clades of the phylogenetic tree and its topology suggest a spatial and temporal sequence of phylogenetic separations that coincide with some major paleogeographic separations during the geological history of the Aegean Sea. The results stress the need for a reconsideration of the evolutionary history of Balkan Podarcis species and help overcome difficulties that classical taxonomy has encountered at both the species and subspecies level.


Spectrofluorimetric determination of terbinafine hydrochloride and linezolid in their dosage forms and human plasma.
A highly sensitive, simple and rapid spectrofluorimetric method was developed for the determination of Terbinafine HCl (TRH) and linezolid (LNZ) in their pharmaceutical formulations. The proposed method is based on measuring the native fluorescence of the studied drugs in water at 336 nm after excitation at 275 nm for TRH and 375 nm after excitation at 254 nm for LNZ. The fluorescence-concentration plots were rectilinear over the range of 0.02-0.15 μg/mL for TRH and 0.5-5.0 μg/mL for LNZ. With lower detection limits of 3.0 and 110.0 ng/mL and a lower quantification limit of 9.0 and 320.0 ng/mL for TRH and LNZ, respectively. The method was successfully applied to the analysis of TRH in its commercial tablets, cream, gel and spray formulations and the results were in good agreement with those obtained with the official method. In addition the method was also applied to the analysis of LNZ in its capsule and I.V solution and the results were in good agreement with those obtained with the comparison method. The effect of sensitizers was studied. The method was extended to the determination of the studied drugs in spiked human plasma and the results were satisfactory.


COMPARATIVE EVALUATION OF SKIN SUTURE IN RATS WITH POLYGLYCAPRONE 25 AND NYLON.
Currently, the market offers a wide variety of suture threads, made of materials with different structural and chemical properties. Among many other characteristics, they vary in origin, absorption or degradation, and structure. From this variety, the clinical doubt arises as to which material provides the patient with the best healing quality. This study aims to comparatively evaluate two different types of suture threads-Monocryl® (polyglycaprone 25) and Ethilon® (nylon)-regarding their ability to aid in tissue regeneration by a histological and immunohistochemical analysis of the skin of rats sutured with the aforementioned materials. This basic experimental study used 12 adult Wistar rats, randomly divided into three groups with four animals each and subjected to four longitudinal incisions under anesthesia. Each group corresponded to a postsurgical evaluation date (one, seven, and 14 days). At 14 postoperative days, the studied groups had no histological difference. However, the use of nylon thread showed greater evidence of earlier fibrotic union. This study found no histological difference in healing 14 days after surgery among the techniques and the types of suture threads. Level of Evidence II, Therapeutic Studies.


The impact of metabolic syndrome on long-term outcomes of percutaneous nephrolithotomy (PCNL).
Study Type--Prognosis (cohort) Level of Evidence 2b. What's known on the subject? and What does the study add? The presence of metabolic syndrome is associated with development of kidney stones and an increase in the stone-recurrence rate. However, studies reporting long-term results of percutaneous nephrolithotomy (PCNL) in metabolic syndrome are lacking. The present study showed that metabolic syndrome was associated with worsening renal function at long-term follow-up and the stone-recurrence rate recurrence after PCNL in patients with metabolic syndrome was 3.2-fold higher compared with the control group. • To investigate the impact of metabolic syndrome on long-term kidney function and stone recurrence rates after percutaneous nephrolithotomy (PCNL). • In all, 73 patients with metabolic syndrome who underwent stone analysis and had a minimum follow-up of 12 months after PCNL were reviewed. • In addition, 73 patients without any metabolic syndrome components who had undergone PCNL and were followed-up for at least 12 months were included in the study as the control group. • These control group patients were selected from 226 patients who were matched with the patients with metabolic syndrome using a 1 : 1 ratio. The matching parameters were age, gender and stone size. • Stone analyses showed that calcium oxalate monohydrate (52.0%) and uric acid (21.9%) were most common among patients with metabolic syndrome, whereas calcium oxalate monohydrate (76.7%) was the most common stone type in the control group. • Stone recurrences occurred with a mean (sd, range) of 36.1 (21.3, 12-109) months follow-up in 26 patients (41.9%) and 12 patients (18.9%) in the metabolic syndrome and control groups, respectively (P = 0.003). • While estimated glomerular filtration rate was decreased from 87.8 to 66.6 mL/min/1.73 m(2) in the metabolic syndrome group, it changed from 96.4 to 91.2 mL/min/1.73 m(2) in control group at long-term follow-up. • The most frequent stone type was calcium oxalate monohydrate in patients with or without metabolic syndrome. • In patients with metabolic syndrome who underwent PCNL, the stone recurrence rate was >40%. • Metabolic syndrome is associated with worsening renal function at long-term follow-up.


Pioglitazone Induces Dysplastic Urothelial Changes in Urinary Bladder of Experimental Diabetes.
Pioglitazone (PIO) is a widely prescribed oral antidiabetic drug that has concerns regarding a potential risk of developing carcinoma of the urinary bladder. The objective of the current study was to assess this potential risk. The potential risk of PIO-induced urinary bladder carcinoma was assessed in the current study by examining urinary bladder of rats for urothelial cytokeratin (CK) expression and proliferative activity by Ki67 immunostaining. Histological examination revealed dysplastic urothelial changes in PIO per se and diabetes mellitus + PIO (diabetic rats receiving PIO). In addition, a significantly (P < 0.05) decreased CK7 and CK8 expression together with a significantly increased CK20 as well as Ki67 expression was detected in the urothelial cells of groups administrated PIO, contrary to those which did not. The manifestations of urothelial dysplasia evidenced by histological examination as well as by the aberrant expression in CK and Ki67 after PIO administration add supporting evidence at cellular and experimental level to the previous clinical suspicions.


Pneumoencephalographic changes with intrasellar cisternal herniation (primary empty sella).
In a selected material of 47 cases of primary intrasellar cisternal herniation, analysis of pneumoencephalographic changes demonstrated the long axis of the anterior part of the third ventricle to point towards the center of the herniation. Transmitted CSF pulsations from the third ventricle probably determine the formation and location of intrasellar herniations. The distribution of air in the subarachnoid space indicated a disturbed CSF circulation in the majority of patients. Visual field defects were unrelated to the slight deformity of the optic nerves observed but rather to a high position of the third ventricle.


Cellular inactivation induced by a radiopharmaceutical kit: role of stannous chloride.
Stannous chloride (SnCl2) has been used in many sectors of human activities such as food manufacturing and in nuclear medicine to produce radiopharmaceuticals labeled with technetium-99m (99mTc). Due to its importance and genotoxic potentiality, we decided to evaluate the biological effect induced by a nuclear medicine kit, which includes SnCl2, in association with glucoheptonic acid (GHA) which is employed for brain and renal scintigraphies. These studies were carried out with the Escherichia coli AB1157 strain and the deoxyribonucleic acid (DNA) plasmid pUC 9.1. The experiments, with different concentrations of SnCl2 and GHA, show an inverse relationship between both agents. When the GHA concentration was increased, the cellular inactivation induced by SnCl2 was reduced, as measured by the number of viable cells. Moreover, GHA protects the DNA molecule against the damage induced by SnCl2.


The immunophenotype of acute myeloid leukemia: is there a relationship with prognosis?
Immunophenotyping of acute myeloid leukemia has controversial implications with regards to prognosis. Many associations have been described between individual antigen expression on myeloid blasts and prognosis, however few are consistent. Markers with a consistent prognostic association that have been demonstrated in more than one study have been CD13, CD14, and CD15. The association of the expression of CD11b with poor prognosis appears definite, as does CD7 expression. When compared with the expression of a single antigen, a pattern of antigen expression is likely to have a more significant relationship to prognosis. This is exemplified by the panmyeloid phenotype (expression of 5 myeloid antigens) which appears to be associated with a good prognosis and may differentiate a subgroup within an otherwise intermediate prognosis group of patients. Further analysis with the inclusion of novel antibodies and the combination of multiple antibodies to create further subgroups such as the panmyeloid phenotype will continue to enhance knowledge in this area.


Primary epithelial tumors of the bladder in children.
Epithelial tumors of the bladder in children are extremely rare and recurrences are even more unusual. We report 3 cases of which 2 had recurrence. ABO(H) antigens were negative in 1 case and they were mildly positive in the other. Although the current opinion is that epithelial tumors in children are benign and single, with a low incidence of malignancy, 3 other pediatric cases of epithelial tumor recurrence have been reported in the literature. Therefore, we wish to emphasize the importance of long-term and careful followup of these patients, since further recurrences are possible.


Concave polyarenes with sulfide-linked flaps and tentacles: new electron-rich hosts for fullerenes.
Pentakis(1,4-benzodithiino)corannulene (6) in CS2 formed the strongest 1:1 complexes with C60 and C70 of any corannulene derivative yet reported. The 1,3,5,7,9-pentakis(propylthio)corannulene (4b) formed weaker 1:1 complexes with C60 and C70 in CS2, whereas the decakis(propylthio)corannulene (5b) and unsubstituted corannulene (1) showed no evidence for complexation with either C60 or C70 in CS2.


In situ enzymatic activity of transglutaminase isoforms on brain tissue sections of rodents: A new approach to monitor differences in post-translational protein modifications during neurodegeneration.
Mammalian transglutaminases (TGs) catalyze the irreversible post-translational modifications of proteins, the most prominent of which is the calcium-dependent formation of covalent acyl transfers between the γ-carboxamide group of glutamine and the ε-amino-group of lysine (GGEL-linkage). In the central nervous system, at least four TG isoforms are present and some of them are differentially expressed under pathological conditions in human patients. However, the precise TG-isoform-dependent enzymatic activities in the brain as well as their anatomical distribution are unknown. Specificity of the used biotinylated peptides was analyzed using an in vitro assay. Isoform-specific TG activity was evaluated in in vitro and in situ studies, using brain extracts and native brain tissue obtained from rodents. Our method allowed us to reveal in vitro and in situ TG-isoform-dependent enzymatic activity in brain extracts and tissue of rats and mice, with a specific focus on TG6. In situ activity of this isoform varied between BACHD mice in comparison to their wt controls. TG isozyme-specific activity can be detected by isoform-specific biotinylated peptides in brain tissue sections of rodents to reveal differences in the anatomical and/or subcellular distribution of TG activity. Our findings yield the basis for a broader application of this method for the screening of pathological expression and activity of TGs in a variety of animal models of human diseases, as in the case of neurodegenerative conditions such as Huntington׳s, Parkinson׳s and Alzheimer׳s, where protein modification is involved as a key mechanism of disease progression.


Microplastics presence in cultured and wild-caught cuttlefish, Sepia officinalis.
Amongst cephalopods microplastics have been reported only in jumbo squid gut. We investigated microplastics in the digestive system of wild cuttlefish (Sepia officinalis) as they are predators and prey and compared the stomach, caecum/intestine and digestive gland (DG) of wild and cultured animals, exposed to seawater from a comparable source. Fibers were the most common type (≈90% of total count) but were ≈2× higher in relation to body weight in wild vs. cultured animals. Fibers were transported to the DG where the count was ≈2× higher /g in wild (median 1.85 fibers/g) vs. cultured. In wild-caught animals the DG was the predominant location but in cultured animals the fibers were more evenly distributed in the digestive tract. The potential impact of microplastics on health of cuttlefish is discussed. Cuttlefish represent a previously unrecognized source of microplastic trophic transfer to fish and finding fibers in cultured animals has implications for aquaculture.


The anomalous origin of the left coronary artery from the right aortic sinus: is the coronary angiography still a 'gold standard'?
Coronary artery anomalies remain a poorly understood topic in modern cardiology. The most important issue is the origin of the left coronary artery or the left anterior descending artery from the opposite aortic sinus, frequently associated with sudden cardiac death. We report our experience concerning the evaluation of these anomalies. From 15 April 1997 to 1 December 2004, we performed 13.407 coronary angiographies and found eight patients with these anomalies. In seven patients the coronary angiography was sufficient for the ultimate decision. However, in one case was the angiographic signs contradictory and the optimal imaging of the coronary tree was received by the multi-slice spiral computer tomography. We consider the coronary angiography a sufficient method of evaluation in most of the patients with the coronary artery anomalies, but the 'gold standard' is 3-dimensional examination by the multi-slice computer tomography or the magnetic resonance. The computer tomography is the method of the choice to distinguish interarterial, intraseptal and prepulmonary course of the left coronary artery originating from the right aortic sinus.


The National Weight Control Registry: a critique.
This article is a critique of the claim that the National Weight Control Registry provides data showing that a significant number of adults in the United States have achieved permanent weight loss. We believe that promoting calorie-restricted dieting for the purpose of weight loss is misleading and futile. We advocate the adoption of a health-at-every-size (HAES) approach to weight management, focusing on the achievement and maintenance of lifestyle changes that improve metabolic indicators of health.


Water and protein structure in photoaged and chronically aged skin.
Changes in the structural proteins and hydration during aging is responsible for altered skin morphologic and mechanical properties manifested as wrinkling, sagging, loss of elasticity, or apparent dryness. To gain insight into the age-related alterations in protein conformation and water structure, we obtained Raman spectra from the sun-protected buttock skin representing chronologic aging and the sun-exposed forearm skin representing combined effects of photoaging and chronologic aging. Ten aged individuals (five men, five women; age range 74-87) and 10 control young individuals (five men, five women; age range 22-29) entered the study. In the photoaged forearm skin the positions of protein-specific amide I, amide III, and CH stretching bands were shifted, suggesting increased protein folding. In contrast, major changes were seen only in the amide I peak in chronologically aged skin. The intensity of the 3250 cm(-1) OH stretching band was increased in photoaged skin (but not in chronologically aged skin) indicating an increased water content. R(v) representation of the low-frequency region of Raman spectra was applied to determine water structure. In the young skin and chronologically aged skin water was mostly present in the bound form. In the photoaged skin, however, an increase in intensity at 180 cm(-1) was noted, which reflects an increase in the not-protein bound water (tetrahedron water clusters). In conclusion, it seems that proteins in the photoaged skin are more compact and interact with water to limited degree. Impairment in protein hydration may add to the understanding of ultrastructural, mechanical, and biochemical changes in structural proteins in the aged skin.


Are breast tumours resistant to tamoxifen also resistant to pure antioestrogens?
A substantial proportion of patients with breast cancer are treated with the antioestrogen tamoxifen. As with other endocrine therapies, clinical experience has shown that some tumours in which growth is initially attenuated by tamoxifen treatment become resistant to continued drug treatment and resume growth. The mechanisms underlying the development of tamoxifen resistance have yet to be described but represent an important focus of research with the aim of defining what other therapies might be effective following tamoxifen treatment. Secondly, an understanding of tamoxifen resistance might suggest means to develop more effective agents for primary treatment of the disease. The development of pure antioestrogens, for example ICI 164,384 and ICI 182,780, which differ pharmacologically from tamoxifen in being entirely free of oestrogen partial-agonist activity, together with cell and animal models of tamoxifen resistant human breast cancer, has revealed one mechanism which might be of considerable clinical significance. Pure antioestrogens were shown to inhibit the proliferation of a greater proportion of tumor cells than tamoxifen in vitro, a differential effect that was attributed to the oestrogenic activity of tamoxifen. Subsequently, cell culture studies have shown that breast cancer cell lines selected for resistance to tamoxifen can still remain sensitive to the growth inhibitory action of pure antioestrogens. Similarly, the growth of human breast tumours in nude mice, which is initially attenuated by tamoxifen but then resumes, can be inhibited by pure antioestrogens. Both types of experiment are consistent with the view that tamoxifen resistance in these model systems is due to the oestrogenic action of tamoxifen. Thus, it can be predicted that in some patients whose tumours recur during tamoxifen therapy, a further response to pure antioestrogen treatment might occur. Studies to examine this hypothesis are currently being undertaken with ICI 182,780. One mechanism which might account for the experimental observations is an intrinsic heterogeneity amongst breast tumour cells in their response to tamoxifen, i.e. that there are at least two different populations of cells; one population which responds to tamoxifen as an antioestrogen and one which "reads" tamoxifen as an oestrogen. The growth advantage thus conferred on the latter population would lead to its predominance. If this is what actually happens in a proportion of human tumours, it can be argued that primary treatment of the tumour with a pure antioestrogen, rather than tamoxifen, would be preferred since a more complete and longer-lasting response would be predicted. Recent comparative studies with human breast tumours grown in nude mice support these predictions.


A critical role for DNA end-joining proteins in both lymphogenesis and neurogenesis.
XRCC4 was identified via a complementation cloning method that employed an ionizing radiation (IR)-sensitive hamster cell line. By gene-targeted mutation, we show that XRCC4 deficiency in primary murine cells causes growth defects, premature senescence, IR sensitivity, and inability to support V(D)J recombination. In mice, XRCC4 deficiency causes late embryonic lethality accompanied by defective lymphogenesis and defective neurogenesis manifested by extensive apoptotic death of newly generated postmitotic neuronal cells. We find similar neuronal developmental defects in embryos that lack DNA ligase IV, an XRCC4-associated protein. Our findings demonstrate that differentiating lymphocytes and neurons strictly require the XRCC4 and DNA ligase IV end-joining proteins and point to the general stage of neuronal development in which these proteins are necessary.


Continuous prospectively navigated multi-echo GRE for improved BOLD imaging of the kidneys.
The objective of this study is to develop improved methods for renal blood oxygenation level dependent (BOLD) imaging. T2* mapping of the kidneys, or renal BOLD imaging, may depict renal oxygen levels and may be valuable as a noninvasive means of following the progression of renal disease. Current renal BOLD data is limited by imaging in a single breath hold, which results in low resolution and low signal-to-noise ratio (SNR). We compare a new free-breathing renal BOLD method with conventional breath-hold BOLD (BH-BOLD). A multi-echo GRE sequence with continuous prospective respiratory navigation and real-time feedback was developed that allows high resolution and high SNR renal BOLD imaging with constant sequence repetition time (TR) during free-breathing BOLD (FB-BOLD). The sequence was evaluated in 10 normal volunteers and compared with conventional BH-BOLD. Scan time for the FB-BOLD sequence was approximately three minutes, compared with 15 seconds for the BH-BOLD sequence. SNR of source images and residual error of T2* fitting were compared between the two methods. The FB-BOLD sequence produced motion-free T2* maps of the kidneys with SNR 1.9 times higher than BH-BOLD images. Residual error of T2* fitting was consistently lower in the right kidney with FB-BOLD (30% less than BH-BOLD) but higher in the left kidney (80% more than BH-BOLD), likely related to placement of the navigator on the right hemidiaphragm. A free-breathing prospectively navigated renal BOLD sequence allows flexible tradeoff between scan time, resolution, and SNR.


Gas chromatographic-mass spectrometric characterisation of amiton and the recovery of amiton from concrete, paint, rubber and soil matrices.
Amiton [O,O-diethyl S-[2-(diethylamino)ethyl] phosphorothiolate], is an organophosphorus chemical included in Schedule 2 of the Chemical Weapons Convention (CWC). Verification provisions under the CWC rely on the existence of a database of analytical information for scheduled chemicals and related compounds. Little analytical information is available for amiton. In this study, gas chromatography-mass spectrometry (GC-MS) characterisation of amiton and its typical impurities (including by-products and degradation products), supported by selective GC detection and 31P NMR data, was undertaken. Twenty-one compounds, including a by-product unique to amiton from an industrial source, were identified. Involatile degradation products of amiton were derivatised to enable their identification by GC-MS. The recovery of amiton from matrices that may be expected in an inspection scenario (i.e. concrete, paint, rubber and soil) was also examined. Paint and concrete matrices were the most useful matrices for the detection of amiton, and its by-products and degradation products. Amiton was readily detected in these matrices after 28 days.


The optimal immune checkpoint inhibitors combined with chemotherapy for advanced non-small-cell lung cancer: a systematic review and meta-analysis.
Immune checkpoint inhibitors (ICIs) plus chemotherapy (CT) have strikingly expanded the therapeutic landscape for advanced non-small cell lung cancer (NSCLC), but little is known about which is superior. We performed a meta-analysis that compared the efficacy and safety of PD-1 inhibitor + CT with PD-L1 inhibitor + CT. PubMed, Embase, Web of Science, Cochrane Library, and major international scientific meetings were searched for relevant randomized controlled trials (RCTs), and the indirect analysis was performed for PD-1 + CT vs PD-L1 + CT. The outcomes included progression-free survival (PFS), overall survival (OS), objective response rate (ORR) and treatment-related adverse events (TRAEs). 8 phase III RCTs with 4253 patients comparing PD-1/PD-L1 + CT in NSCLC were included. The PD-1 + CT led to notably longer OS most in low/negative expression of PD-L1 for NSCLC patients compared with PD-L1 + CT. In terms of Grade 3-5 TRAEs, the results showed that PD-1 + CT and PD-L1 + CT exclusively increased the risk of adverse incidence than CT alone, especially for PD-L1 + CT (p < 0.00001). For subgroups including female, young patients, patients with nonsmoker, and EGFR/ALK wild-type, PD-1 + CT was associated with prolonged OS (p < 0.05). Meanwhile, for no liver metastasis of NSCLC patients, we found obviously OS advantage for patients treated with PD-1 + CT compared to PD-L1 + CT. ICIs + CT seemed to be more effective first-line regimen and PD-1 + CT could be recommended as the first-rank therapy for advanced NSCLC patients with low/negative expression of PD-L1. However, we should be particularly vigilant about the occurrence of the Grade 3-5 TRAEs.


The small nuclear ribonucleoproteins that react with anti-Sm and anti-RNP antibodies.
We have examined the polypeptide pattern of small nuclear ribonucleoprotein particles that react with monoclonal anti-Sm antibodies or polyclonal anti-(U1)RNP antibodies. The fresh nuclear extracts analyzed were prepared from human cells that had been pulse-chased with a mixture of 15 3H-labeled amino acids. In contrast to previous reports in the literature, the apparent molecular weights of the major polypeptides that remained in the 1 M NaCl-washed ribonucleoprotein-antibody complexes were approximately 80000, 55000, 28000, 25000, 14000 and 9000, when probed with monoclonal anti-Sm antibodies, and about 69000, 58000 and 35000, when polyclonal anti-(U1)RNP antibodies were used.