Heat-shock protein 90α is a potential prognostic and predictive biomarker in hepatocellular carcinoma: a large-scale and multicenter study.
Although the diagnostic value of plasma heat-shock protein 90α (HSP90α) in hepatocellular carcinoma (HCC) has been previously reported, the causal effect of the plasma HSP90α levels on HCC prognosis remains largely unclear. To this extent, we sought to assess whether the plasma HSP90α acts as a prognostic factor for HCC patients. A total of 2150 HCC patients were included in this retrospective study between August 2016 and July 2021. Plasma HSP90α levels were tested within a week before treatment and their association with prognosis was assessed. An optimal cutoff value of 143.5 for the HSP90α based on the overall survival (OS) was determined using the X-tile software. HCC patients with HSP90α < 143.5 ng/mL (low HSP90α) before and after propensity score matching (PSM) indicated longer median OS (mOS) relative to those with HSP90α ≥ 143.5 ng/mL (high HSP90α) (37.0 vs. 9.0 months, p < 0.001; 19.2 vs. 9.6 months, p < 0.001; respectively). In addition, the high HSP90α plasma level is an independent poor prognostic factor for OS in HCC patients. In our subgroup analysis, including the supportive care group, surgery group, transarterial chemoembolization (TACE) group, adjuvant TACE group, an immune checkpoint inhibitor (ICI) plus targeted therapy group, and TACE plus ICI group, the high HSP90α group demonstrated better OS compared to the low HSP90α group. Moreover, in the supportive care, TACE, ICI plus targeted therapy, TACE plus ICI groups, and high HSP90α levels were also an independent poor prognostic factors for OS. Our study confirmed that the plasma HSP90α level can be used as a prognostic biomarker for HCC.


Association of HLA-DR3 with an increase in peripheral blood lymphocytes.
Normal healthy individuals that were HLA-DR3 positive were found to have significantly higher total lymphocyte counts in peripheral blood compared to healthy individuals that were HLA-DR3 negative.


Dihydroartemisinin inhibits the tumorigenesis and invasion of gastric cancer by regulating STAT1/KDR/MMP9 and P53/BCL2L1/CASP3/7 pathways.
Dihydroartemisinin (DHA), an effective antimalarial drug, has been widely investigated as an anti-tumor agent. Although previous studies have indicated the potential therapeutic effects of DHA on multiple malignancies, its detailed molecular mechanisms in gastric cancer (GC) are still undocumented. In the present study, we applied network pharmacology and bioinformatics (gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses) to obtain the collective targets of DHA and GC and analyzed their involvement in constructing a protein-protein interaction (PPI) network. The top 10% hub targets in this network were identified, and TCGA database was utilized for the single gene analysis of their correlation with the prognosis of GC. CCK8, EdU, Transwell, and flow cytometry analyses were conducted, and subcutaneous xenograft tumor models were constructed to assess the effects of DHA on the tumorigenesis and invasion of GC. Furthermore, the targets of DHA were verified by molecular docking, quantitative real-time PCR (qPCR) and western blot analyses in GC cells. The results indicated that the common targets of DHA and GC were enriched in multiple cancer-related pathways including KDR, STAT1 and apoptosis signaling pathways, where the core genes included KDR, MMP9, STAT1, TP53, CASP3/7 and BCL2L1. The lowered expression of KDR and increased expression of TP53 and CASP7 harbored a favorable survival for patients with GC patients. CASP7 showed a positive correlation with CASP3 but a negative correlation with KDR and could be regarded as an independent protective factor for overall survival in GC. Moreover, DHA treatment induced cell apoptosis and suppressed the cell proliferation, DNA synthesis, cycle progression and invasive capabilities both in vitro and in vivo. DHA also upregulated p53, CASP3, and cleaved-CASP3 and downregulated BCL2L1, MMP9, KDR, p-KDR, STAT1 and p-STAT1 in GC cell lines. In conclusion, DHA could suppress the tumorigenesis and invasion of GC by regulating STAT1/KDR/MMP9 and p53/BCL2L1/CASP3/7 pathways. Our findings might provide a novel approach for the treatment of GC.


Space-time clustering of elevated thyroid stimulating hormone levels.
Previous studies of congenital hypothyroidism (CHT) have reported an increasing incidence which may suggest that environmental factors play an aetiological role. If so, then cases may exhibit space-time clustering, where cases occur at similar times and close proximities to other cases. In this study we investigated whether space-time clustering of elevated thyroid stimulating hormone (TSH) in newborns exists. All infants born in the Northern Region of England are screened by measuring levels of circulating TSH using a blood spot assay. Data on 207 cases of elevated TSH values, as a proxy for CHT, in newborns born from 1994 to 2006 inclusive were available and analysed using rigorous space-time clustering statistical methods. Analysis showed statistically significant evidence of space-time clustering. The strength of clustering was most marked for cases born within 0.1-0.7 year (1-8 months) of one another. This is the first study to find significant space-time clustering of cases of elevated TSH levels in newborns, a surrogate for space-time clustering of CHT. Whilst the reasons for the clustering are unclear, it would appear from this analysis that transient environmental exposures are likely to be involved, although environmental determinants of genetic mutations and epigenetic factors cannot be ruled out. Further research is required to a) validate these results in other populations and b) to assess in more detail the potential environmental determinants of increased CHT risk.


Enhancement of phagocytosis in mouse macrophages by pituitary adenylate cyclase activating polypeptide (PACAP) and related peptides.
The effects of pituitary adenylate cyclase activating polypeptide (PACAP) and related peptides on phagocytosis of fluorescent latex beads by mouse peritoneal macrophages were examined using flow cytometry (FCM). PACAP38, PACAP27 and vasoactive intestinal peptide (VIP) enhanced phagocytosis in a dose-dependent manner. Relative potencies of related peptides at a concentration of 10(-6) M were PACAP38 > PACAP27 > VIP > secretin > glucagon > (peptide with NH2-terminal histidine and COOH-terminal methionine amide, in short PHM). PACAP(6-38) was as effective as PACAP38. PACAP(6-27) enhanced phagocytosis more effectively than did PACAP27. PACAP(28-38) slightly enhanced phagocytosis. The present result suggest that PACAP38 is one of the mediators that the nervous system uses to modulate the immune system.


TNF-alpha is a mediator of the anti-inflammatory response in a human neonatal model of the non-septic shock syndrome.
The anti-inflammatory/immunoparalytic phase of the systemic inflammatory response syndrome (SIRS) following major insult (surgery, thermal/traumatic injury) is of major clinical importance in the neonate, during which the risk of infection is particularly great. Here, the mechanisms by which TNF-alpha production is suppressed in response to infection are largely unknown. We questioned whether TNF-alpha itself could be a critical mediator of this suppression. Monocytes, isolated from cord blood (n=3), were treated with LPS (100 ng/ml), TNF-alpha (10 ng/ml, +/- anti-TNF-alpha antibody) for 18 and 36 h. Cells were then restimulated with LPS (Gram -ve) or Pam-3-Cys (Gram +ve) for 24 h. This was also done in the presence of selective inhibitors of MAP kinases p38, MEK and JNK. TNF-alpha, IL-6, IL-10 and IL-8 were quantified by ELISA CD86 and HLA-DR expression were determined flow cytometrically. Cells stimulated with LPS for 24 h produced TNF-alpha (282 pg/ml), IL-10 (1,236 pg/ml), IL-6 (2,694 pg/ml) and IL-8 (2,144 pg/ml). In cells pre-exposed to TNF-alpha for 36 h, there was a significant suppression in TNF-alpha and IL-6 levels (9 and 221 pg/ml, respectively) (P<0.05) with minimal impact on IL-10 (1,206 pg/ml) and IL-8 levels (1,886 pg/ml). A similar effect was seen with Pam-3-Cys with a tenfold decrease in levels of TNF-alpha and IL-6 (86-->8.5 pg/ml and 458-->46 pg/ml, respectively) with no effect on IL-10 and IL-8 levels. Anti-TNF-alpha antibody negated this effect. Inhibition of p38 kinase reversed the TNF-alpha effect. Inhibition of the JNK and MEK kinases had no effect. A reduction in the expression of CD86 and HLA-DR was observed. This ex-vivo model of non-septic SIRS demonstrates that TNF-alpha, released during a major insult, can suppress subsequent monocyte responses to bacterial agents through p38 MAP kinase, making it a potential therapeutic target.


Comparative evaluation and selection of a method for lipid and fatty acid extraction from macroalgae.
A comparative evaluation of Bligh and Dyer, Folch, and Cequier-Sánchez methods for quantitative determination of total lipids (TLs) and fatty acids (FAs) was accomplished in selective green (Ulva fasciata), red (Gracilaria corticata), and brown algae (Sargassum tenerrimum) using a full factorial categorical design. Applications of sonication and buffer individually on lipid extraction solvent systems were also evaluated. The FA recoveries obtained from the aforementioned methods were compared with those of direct transesterification (DT) methods to identify the best extraction methods. The experimental design showed that macroalgal matrix, extraction method, and buffer were key determinants for TL and FA recoveries (P≤0.05), exhibiting significant interactions. But sonication gave erratic results with no interaction with any of the factors investigated. The buffered solvent system of Folch rendered the highest TL yield in U. fasciata and G. corticata while the buffered system of Bligh and Dyer gave the highest yield in S. tenerrimum. DT methods were more convenient and accurate for FA quantification and rendered 1.5-2 times higher yields when compared with the best conventional method, minimizing the use of chlorinated solvents, their cost of analysis, and disposal. The buffered solvent system was found to be the most appropriate for lipid research in macroalgae.


Intracellular stability of the gene encoding influenza virus haemagglutinin.
The intracellular stability of the genome of noninfectious uv-irradiated influenza virus (A/WSN:H1N1) in dividing MDCK cells was investigated using marker rescue techniques. The haemagglutinin gene could still be rescued by infection with A/X49 (H3N2) at 5 weeks postinoculation; its half-life was 13 days.


Spinal ossifying lipoma.
A case of ossifying spinal lipoma is reported. The patient presented with a large swelling over the lumbar region with a dermal sinus in the centre. Imaging revealed an intradural lipoma connected with a large subcutaneous lipoma in which there was a prominent island of bone. The lipoma was excised and there were no postoperative neurological deficits. To the best of our knowledge, this is the first report of a spinal ossifying lipoma associated with a dermal sinus.


Synthesis and sequence-specific proteolysis of a hybrid protein (colicin A::growth hormone releasing factor) produced in Escherichia coli.
DNA constructs coding for human growth hormone (hGH)-releasing factor (hGRF) preceded by the specific recognition sequence for the activated blood coagulation factor X (FXa), fused in frame to the N-terminal 172-amino acid residues of colicin A, have been expressed in Escherichia coli. The construct was placed under the control of the inducible caa promoter in an operon containing a downstream gene coding for the cell lysis protein, Cal. Induction resulted in excretion of only the processed colicin A fragment. Replacement of Cal by the terminator from phage fd resulted in high expression of the hybrid protein, which was recovered as cytoplasmic aggregates. Enzymatic cleavage of the purified and renatured hybrid protein using FXa allowed the recovery of authentic hGRF.


A novel prognostic marker in acute ischemic stroke: small pericardial effusion.
The study aimed to evaluate the prognostic importance of small pericardial effusion (SPE) found on echocardiography in a cohort of patients hospitalized for acute ischemic stroke. We prospectively followed a series of 408 consecutive first-ever acute ischemic stroke patients aged ≥50 years who were admitted to the hospital within 24 h of the onset of stroke symptoms. All of the patients underwent transthoracic echocardiography within the first 48 h. Exclusion criteria were cardiothoracic surgery or acute myocardial infarction within the previous 6 months, a moderate or greater pericardial effusion (>1 cm if circumferential), and inadequate visualization of the pericardial space. The patients were followed for 1 year or until death, whichever came first. SPE was noted in 64 (15.7 %) of the patients. Mortality at 1 year was greater for patients with a small effusion (n = 21, 32.8 %) compared to those without an effusion (n = 40, 11.6 %, p < 0.001). After adjustment for age, demographics, medical history, and other echocardiographic findings, SPE remained associated with higher mortality (OR 2.515; 95 % CI 1.188-5.477; p = 0.008). This study is the first to demonstrate that the presence of SPE is associated with increased mortality in patients with first-ever acute ischemic stroke.


Enhancement of arachidonic acid liberation by protein kinase C activator is partially dependent on extracellular Na+ in rabbit platelets.
In [3H]arachidonic acid-labeled rabbit platelets, pretreatment with phorbol 12-myristate 13-acetate (20 nM) or dioctanoylglycerol (20 microM) enhanced [3H]arachidonic acid liberation induced by low concentration of A23187 (150 nM). When extracellular Na+ was replaced with N-methyl-D-glucamine, the enhancement is reduced by about 50%. Similar synergistic enhancement of the liberation was obtained by using monensin (2-10 microM) or NH4Cl (5-20 mM) in place of protein kinase C activator in combination with A23187. The guanosine 5'-O-[3-thiotriphosphate] (100 microM)-induced liberation was also enhanced by a rise of extracellular pH (pH 7.0-7.8) in saponin-permeabilized platelets. These results suggest that the enhancement of arachidonic acid liberation by protein kinase C may partially be mediated by intracellular alkalinization in rabbit platelets.


Urinary γ-hydroxybutyrate concentrations in 1126 female subjects.
γ-Hydroxybutyrate (GHB) and its metabolic precursor γ-butyrolactone (GBL) are often implicated in cases of drug-facilitated sexual assault (DFSA), although definitive confirmation of GHB/GBL ingestion is complicated by GHB's endogenous nature and rapid elimination following ingestion. Multiple studies have attempted to establish a discriminant limit (generally 10 mg/L) above which urinary GHB concentrations can be considered consistent with GHB/GBL consumption. To supplement the currently available data, a rapid gas chromatography-mass spectrometry method was developed and validated for the analysis of GHB (following acidic conversion to GBL) and used to analyze urine samples collected from 1126 women (mean = 0.84 mg/L, median = 0.68 mg/L, range = 0.00-5.5 mg/L). GHB concentrations were shown to be independent of urinary pH (within the range 4.6-9.3), age (within the range 18-35 years), body mass index (within the range 13.8-36.3), and race. Adjusting GHB concentrations with respect to urinary specific gravity had little effect on the mean value (0.91 mg/L) and range (0.0-7.76 mg/L), although a statistically significant trend of increasing GHB concentration with specific gravity could be observed. Our results can be taken to offer further support for the 10 mg/L discriminant limit for GHB administration in antemortem urine samples.


Upstream Machine Learning in Radiology.
Machine learning (ML) and Artificial intelligence (AI) has the potential to dramatically improve radiology practice at multiple stages of the imaging pipeline. Most of the attention has been garnered by applications focused on improving the end of the pipeline: image interpretation. However, this article reviews how AI/ML can be applied to improve upstream components of the imaging pipeline, including exam modality selection, hardware design, exam protocol selection, data acquisition, image reconstruction, and image processing. A breadth of applications and their potential for impact is shown across multiple imaging modalities, including ultrasound, computed tomography, and MRI.


Does alcohol-related desire thinking predict in-the-moment drinking behaviours?
Desire thinking (voluntary thinking involving a perseverative focusing on desired target-related memories, images, and information), is an important factor in the experience of drinking urges and cravings. Research has not examined the relationship between desire thinking, behavioural intention and the implementation of that behaviour into action, nor how one's past repeated behaviour in situ (expressed as habit) may moderate these relationships. We employed an ad libitum drinking paradigm to provide indirect measures of in-the-moment drinking behaviours (amount poured [mls] and amount consumed [mls]) in a group of eighty-eight self-defined social drinkers immediately after measuring self-reported drinking habit, drinking-related desire thinking, and general drinking behaviour. Results confirmed the predicted positive relationship between desire thinking and in-the-moment drinking behaviour with the effects increasing as a function of rising drinking habit strength. We also observed a dissociation between desire thinking components (verbal perseveration and imaginal prefiguration) in the moderating effects of habit strength on drinking behaviour. For imaginal prefiguration (thoughts related to construction of mental images of a desired target or of its context for consumption) a direct effect on drinking behaviours was shown. In comparison, the effect for verbal perseveration (repetitive self-talk regarding the need to achieve a desired target) was not shown to independently predict drinking-related behaviour but was significantly moderated by increasing drinking-related habit strength. Future work should formulate the nature of this moderating influence on perseverative goal-directed thinking.


A Large Tn7-like Transposon Confers Hyper-Resistance to Copper in Pseudomonas syringae pv. syringae.
Copper resistance mechanisms provide an important adaptive advantage to plant pathogenic bacteria under exposure to copper treatments. Copper resistance determinants have been described in Pseudomonas syringae pv. syringae (Pss) strains isolated from mango intimately associated with 62 kb plasmids belonging to the pPT23A family (PFP). It has been previously described that the indiscriminate use of copper-based compounds promotes the selection of copper resistant bacterial strains and constitutes a selective pressure in the evolution of copper resistance determinants. Hence, we have explored in this study the copper resistance evolution and the distribution of specific genetic determinants in two different Pss mango populations isolated from the same geographical regions, mainly from southern Spain with an average of 20 years of difference. The total content of plasmids, in particular the 62 kb plasmids, and the number of copper resistant Pss strains were maintained at similar levels over the time. Interestingly, the phylogenetic analysis indicated the presence of a phylogenetic subgroup (PSG) in the Pss mango phylotype, mostly composed of the recent Pss population analyzed in this study that was strongly associated with a hyper-resistant phenotype to copper. Genome sequencing of two selected Pss strains from this PSG revealed the presence of a large Tn7-like transposon of chromosomal location, which harbored putative copper and arsenic resistance genes (COARS Tn7-like). Transformation of the copper sensitive Pss UMAF0158 strain with some putative copper resistance genes and RT-qPCR experiments brought into light the role of COARS Tn7-like transposon in the hyper-resistant phenotype to copper in Pss.IMPORTANCECopper compounds have traditionally been used as standard bactericides in agriculture in the past few decades. However, the extensive use of copper has fostered the evolution of bacterial copper resistance mechanisms. Pseudomonas syringae is a plant pathogenic bacterium used worldwide as a model to study plant-pathogen interactions. The adaption of P. syringae to plant surface environment is the most important step prior to an infection. In this scenario, copper resistance mechanisms could play a key role in improving its epiphytic survival. In this work, a novel Tn7-like transposon of chromosomal location was detected in P. syringae pv. syringae strains isolated from mango. This transposon conferred the highest resistance to copper sulfate described to date for this bacterial phytopathogen. Understanding in depth the copper resistance mechanisms and their evolution are important steps to the agricultural industry to get a better improvement of disease management strategies.


Synthesis, Characterization and Fluorescence Properties of Novel Porous Fe/ZnO Nano-Hybrid Assemblies by Using Berberis thunbergii Extract.
In this work, novel Fe/ZnO nanocomposites (NCs) and Fe nanoparticles loaded onto porous ZnO nanostructures have been synthesized via a simple biotechnological route by using Berberis thunbergii extract. In this direction, the as-synthesized bio-based porous ZnO derivatives and human serum albumin (HSA), as a biopolymeric model, form nano-hybrid assemblies. The effect of loading Fe on properties of porous ZnO nanostructures as well as the behavior of the nano-hybrid assemblies were evaluated by using XRD, SEM, EDX, DLS, PL, CD, FTIR and UV/Visible-diffuse reflectance spectra (UV/Vis-DRS) techniques. The fluorescence results revealed that the interaction of Fe/ZnO NCs with HSA biopolymer led to the formation of a ground state complexes as nano-hybrid assemblies. The calculated thermodynamic parameters indicated that the binding process occurred spontaneously. The CD and FTIR spectra confirmed the changes in helicity of HSA as well as the random coil and β-turn in the secondary structure of HSA upon interaction with Fe/ZnO NCs.


Immunohistological differential diagnosis of inflammatory colonic diseases.
Immunohistological investigations were carried out on human colonic tissue from, I healthy mucosa, 2 slightly inflamed mucosa, 3 mucosa with ulcerative colitis, 4 mucosa with Crohn's colitis, using antibodies against immunoglobulins and complement components. All our antibodies, including F(ab')2 fragments, demonstrated a progressive increase of labelled cells from healthy mucosa through slightly inflamed mucosa to mucosa with ulcerative colitis, in contrast to a complete absence of labelled cells in cases of Crohn's disease. The results are discussed with regard to their pathogenesis and their clinical significance for the differentiation of ulcerative colitis and Crohn's colitis.


Computed tomography of hamartoma of the epiglottis.
We present the computed tomography (CT) findings of a hamartoma of the epiglottis occurring in a 58-year-old man. Although hamartomas are particularly uncommon in this site. CT identification of fat within the complex mass facilitated diagnosis without recourse to an invasive biopsy.


Changing appearance of retinal arteriovenous malformation.
Retinal arteriovenous malformations (racemose angiomas) are usually described as non-changing congenital vascular anomalies. The authors describe prominent retinal vascular changes that occurred in the fundus of a patient with a complex retinal arteriovenous malformation as part of the Wyburn-Mason syndrome during a follow-up period of 17 years.


Comprehensive profiling of mRNA polyadenylation in specific cell types in vivo by cTag-PAPERCLIP.
The ability to generate cell-type specific mRNA polyadenylation (pA) maps from complex tissues is crucial for understanding how alternative polyadenylation (APA) is regulated in individual cell types in their physiological environment under different conditions. In this chapter, we discuss cTag-PAPERCLIP, a recently developed method combining the well-established CLIP (crosslinking immunoprecipitation) technique and the Cre-lox system to achieve customized cell-type specific APA profiling from mouse tissue without cell purification or enrichment. In cTag-PAPERCLIP, selective expression of GFP-tagged poly(A) binding protein (PABP-GFP) in the desired cell type is achieved through Cre-mediated activation of a latent knock-in allele of PABP-GFP. Immunoprecipitation of PABP-GFP then allows mRNA 3' end fragments in the desired cell type to be specifically retrieved from ultraviolet (UV)-irradiated whole tissue lysate. The mRNA fragments are subsequently turned into a cDNA library to provide a comprehensive APA map and an mRNA expression profile of the chosen cell type through deep sequencing.


Inhibition by ethanol of noradrenaline output from peripheral sympathetic nerves: possible interaction of ethanol with neuronal receptors.
The sympathetic nerve terminals of the isolated rabbit heart perfused with Tyrode solution were used to study the action of ethanol on the noradrenaline uptake and release. The uptake of exogenous noradrenaline (10 ng/ml) into the sympathetic nerve endings, the noradrenaline output evoked by raising the concentration of potassium ions in the perfusion fluid, and the release in response to electrical stimulation of the nerve axons were inhibited only by lethal concentrations of the alcohol; the concentrations which caused 50% inhibition (IC50) amounted to 760 mM, 830mM, and 1150 mM respectively. However, ethanol at concentrations compatible with moderate intoxication reduced the noradrenaline release in response to activation of the nicotine receptors on the nerve terminals by dimethylphenylpiperazine; the threshold concentration was 36 mM and the IC50 was 129 mM. It is suggested that this effect is due to hydrophobic interaction of the alcohol with receptor proteins, thus inhibiting stimulus formation.


The impact of emergency department overcrowding on resident education.
Few studies have evaluated the effect of Emergency Department (ED) overcrowding on resident education. To determine the impact of ED overcrowding on Emergency Medicine (EM) resident education. A prospective cross-sectional study was performed from March to May 2009. Second- and third-year EM residents, blinded to the research objective, completed a questionnaire at the end of each shift. Residents were asked to evaluate the educational quality of each shift using a 10-point Likert scale. Number of patients seen and procedures completed were recorded. Responses were divided into ED overcrowding (group O) and non-ED overcrowding (group N) groups. ED overcrowding was defined as >2 h of ambulance diversion per shift. Questionnaire responses were compared using Mann-Whitney U tests. Number of patients and procedures were compared using unpaired T-tests. During the study period, 125 questionnaires were completed; 54 in group O and 71 in group N. For group O, the median educational value score was 8 (interquartile range [IQR] 7-10), compared to 8 (IQR 8-10) for group N (p = 0.24). Mean number of patients seen in group O was 12.3 (95% confidence interval [CI] 11.4-13.2), compared to 13.9 (95% CI 12.7-15) in group N (p = 0.034). In group O, mean number of procedures was 0.9 (95% CI 0.6-1.2), compared to 1.3 (95% CI 1-1.6) in group N (p = 0.047). During overcrowding, EM residents saw fewer patients and performed fewer procedures. However, there was no significant difference in resident perception of educational value during times of overcrowding vs. non-overcrowding.


Alterations in G-protein-regulated transmembrane signalling induced in murine myocardium by coxsackievirus B3 infection.
Cardiomyopathy is usually associated with marked alterations in myocardial transmembrane signalling. Although acute viral myocarditis may result in chronic cardiomyopathy in some cases, the possible consequences of viral infection on function of the myocardial signal-transducing complex have not been explored. Therefore, the present study was designed to investigate the G-protein-regulated adenylyl cyclase signalling system in murine myocardium during myocarditis induced by coxsackievirus B3 (CVB3) infection. We examined the functional characteristics of adenylyl cyclase as well as the function and distribution of beta-adrenoceptors, m-cholinoceptors and G-proteins in myocardial plasma membranes isolated from the hearts of mice with acute (7 days pi) or late phase (21 days pi) myocarditis and the obtained results were compared with the corresponding data determined in age-matched controls. While the basal adenylyl cyclase activity was not significantly altered, the ability of forskolin, sodium fluoride and GTP gamma S to activate adenylyl cyclase was lowered by about 20% in samples from virus-infected animals. The level of Gs alpha in myocardial plasma membranes as well as the functional activity of Gs alpha was not affected by viral infection, but the Gi alpha content was increased by about 20%. The total number of beta-adrenoceptors in myocardial plasma membranes increased by about 12-15% due to higher content of the beta 2-adrenoceptor subtype. Although the agonist-binding parameters of beta-adrenoceptors were not significantly altered, the ability of these receptors to mediate stimulation of adenylate cyclase was markedly diminished (by 56-80%). The total number of m-cholinoceptors in samples derived from virus-infected mice increased considerably (by 29-59%) and a significant proportion of the receptors shifted to a higher affinity status, but their ability to transduce agonist signals was impaired. These data are the first to demonstrate that several different sites of the myocardial G-protein-regulated adenylyl cyclase signalling complex are significantly altered in acute as well as in late phase of CVB3-induced myocarditis.


Comparison of the enzyme activities of phosphatidylcholine, phosphatidylglycerol and phosphatidylinositol synthesis in freshly isolated type II pneumocytes and whole lung from the adult rat.
We compared the activities of enzymes of phosphatidylcholine, phosphatidylglycerol and phosphatidylinositol synthesis in whole lung tissue and freshly isolated type II pneumocytes from adult rats. The activities of 1-acylglycerophosphocholine acyltransferase and CDPdiacylglycerol-glycerol-3-phosphate 3-phosphatidyltransferase were 2.9- and 4.4-fold higher, respectively, in type II cell sonicates than in whole lung homogenates. There was little difference between the type II cells and whole lung in the activities of choline kinase, choline-phosphate cytidyltransferase, cholinephosphotransferase, phosphatidate phosphatase, phosphatidate cytidylytransferase or CDPdiacylglycerol-inositol 3-phosphatidyltransferase. Since the type II cell is the source of pulmonary surfactant, and disaturated phosphatidylcholine and phosphatidylglycerol are major components of surfactant, it is of interest that this cell is enriched in the activities of enzymes exclusively involved in the synthesis of these lipids. In view of possible proteolytic damage during isolation we compared freshly isolated type II cells with those cultured for 1 day. The rates of incorporation of [methyl-3H]choline and [2-3H]glycerol into phospholipids, L-[U-14C]phenylalanine into protein and [methyl-3H]thymidine into DNA were the same in the freshly isolated and cultured cells. The composition of the phospholipids synthesized from [2-3H]glycerol and sodium [1-14C]acetate were also the same. The freshly isolated cells were at least 90% pure and did not release significant amounts of lactate dehydrogenase. Since use of freshly isolated cells avoids cell loss during culture they provide an attractive alternative, particularly in studies requiring large amounts of material.


Tinea capitis et barbae caused by Trichophyton tonsurans: A retrospective cohort study of an infection chain after shavings in barber shops.
Tinea capitis is a highly contagious infectious disease caused by dermatophytes. In Central Europe, it is mainly caused by zoophilic dermatophytes, as, for example Microsporum (M) canis or Trichophyton (T) mentagrophytes and increasingly by anthropophilic fungi. T tonsurans was commonly related to the Tinea gladiatorum, where transmission occurred between infected persons or via contaminated floors. Reporting the transmission of this highly contagious dermatophyte for the first time via beard shaving and hairdressing in barber shops in Germany. 18 young male patients developed tinea capitis and/or barbae shortly after shavings of the beard and/or hair in a barber shop. Native, cultural and molecular diagnostics as well as tissue biopsies and resistance tests were performed of skin and hair samples. In all samples, T tonsurans could be identified. The medical history and the clinical picture suggest a transmission through contaminated hairdressing tools. The patients were treated with terbinafine or itraconazole in combination with or exclusively with topical antimycotics. The transmission and a resulting increase in the incidence of infections with T tonsurans may be due to shavings with direct skin contact of insufficiently disinfected hairdressing tools. This path of infection has already been observed in Africa and is now being described for the first time in Germany. Knowledge of the pathogen and its transmission ways are essential to interrupt the chain of infection.


Sphenoid sinus mucocele masquerading as a skull base malignancy.
Mucoceles of the sphenoid sinus are rare. Only occasional reports have appeared in the medical literature since they were first described by Berg in 1889. The sphenoid sinus lies deep within the skull at the crossroads of 13 sensitive structures. The presence of a sphenoid sinus mucocele may be accompanied by a wide variety of signs and symptoms depending upon which of these structures are involved. The majority of these lesions are initially misdiagnosed as malignancies leading to increased patient morbidity and possible mismanagement. We present two cases of sphenoid sinus mucoceles which presented with a variety of neurological signs and symptoms. The radiographic findings were felt to be consistent with skull base malignancies. The diagnosis was corrected only after surgical exploration of the sphenoid sinus revealed a mucocele.


Targeting reverse tetracycline-dependent transactivator to murine mammary epithelial cells that express the progesterone receptor.
Through an established gene-targeting strategy, reverse tetracycline-dependent transactivator (rtTA) was targeted downstream of the murine progesterone receptor (PR) promoter. Mice were generated in which one (PR(+/rtTA)) or both (PR(rtTA/rtTA)) PR alleles harbor the rtTA insertion. The PR(+/rtTA) and PR(rtTA/rtTA) knockins exhibit phenotypes identical to the normal and the progesterone receptor knockout mouse, respectively. Crossed with the TZA reporter, which carries the TetO-LacZ responder transgene, the PR(+/rtTA)/TZA and PR(rtTA/rtTA)/TZA bigenics exhibit doxycycline-induced beta-galactosidase activity specifically in progesterone responsive target tissues such as the mammary gland, uterus, ovary, and pituitary gland. In the case of the PR(+/rtTA)/TZA mammary epithelium, dual immunofluorescence demonstrated that PR expression and doxycycline-induced beta-galactosidase activity colocalized; beta-galactosidase was not detected in the absence of doxycycline. Although both the PR(+/rtTA) and PR(rtTA/rtTA) knockins represent innovative animal models with which to further query progesterone's mechanism of action in vivo, the PR(rtTA/rtTA) mouse in particular promises to provide unique insight into the paracrine mechanism of action, which underpins progesterone's involvement in mammary morphogenesis with obvious implications for extending our understanding of this steroid's role in breast cancer progression.


The unicellular eukaryotic parasite Toxoplasma gondii hijacks the migration machinery of mononuclear phagocytes to promote its dissemination.
Toxoplasma gondii is an obligate intracellular protozoan with the ability to infect virtually any type of nucleated cell in warm-blooded vertebrates including humans. Toxoplasma gondii invades immune cells, which the parasite employs as shuttles for dissemination by a Trojan horse mechanism. Recent findings are starting to unveil how this parasite orchestrates the subversion of the migratory functions of parasitised mononuclear phagocytes, especially dendritic cells (DCs) and monocytes. Here, we focus on how T. gondii impacts host cell signalling that regulates leukocyte motility and systemic migration in tissues. Shortly after active parasite invasion, DCs undergo mesenchymal-to-amoeboid transition and adopt a high-speed amoeboid mode of motility. To trigger migratory activation - termed hypermigratory phenotype - T. gondii induces GABAergic signalling, which results in calcium fluxes mediated by voltage-gated calcium channels in parasitised DCs and brain microglia. Additionally, a TIMP-1-CD63-ITGB1-FAK signalling axis and signalling via the receptor tyrosine kinase MET promotes sustained hypermigration of parasitised DCs. Recent reports show that the activated signalling pathways converge on the small GTPase Ras to activate the MAPK Erk signalling cascade, a central regulator of cell motility. To date, three T. gondii-derived putative effector molecules have been linked to hypermigration: Tg14-3-3, TgWIP and ROP17. Here, we discuss their impact on the hypermigratory phenotype of phagocytes. Altogether, the emerging concept suggests that T. gondii induces metastasis-like migratory properties in parasitised mononuclear phagocytes to promote infection-related dissemination.


A novel technique for reconstruction of the medial patellofemoral ligament in skeletally immature patients.
Habitual or recurrent dislocation of the patella in the skeletally immature patient is a particularly demanding problem since the etiology is frequently multifactorial. The surgical techniques successfully performed in adults with patellar instability may risk injury to an open growth plate if applied to children. We present a technique that preserves femoral and patellar insertion anatomy of medial patellofemoral ligament (MPFL) using a free semitendinosus autograft together with tenodesis to the adductor magnus tendon without damaging open physis on the patellar attachment of MPFL. A 3-cm long longitudinal skin incision is performed 10 mm distal to the tibial tuberosity on the anteromedial side. The semitendinosus tendon is harvested with the stripper. The semitendinosus tendon is placed on a preparation board and cleaned of muscle tissue. The usable part of the tendon should be at least 20 cm long and 4 mm wide. The two free ends of the graft are sutured with Krakow technique. A medial longitudinal incision 2 cm in length is made to expose the MPFL and to abrade the patellar attachment of vastus medialis obliquus. The first patellar tunnel is created with 4.5 mm drill at the mid aspect of the medial patella in the anteroposterior and proximal-distal direction. The drill hole is formed parallel to the articular surface of the center of the patella. The second tunnel is created with 3.2 mm drill and the entry point is localized at the center of the patella. These two tunnels intersect to form a single tunnel. The semitendinosus autograft is run through the bone tunnel in the patella. Double-stranded semitendinosus autograft is placed in the presynovial fatty plane between the second and the third layer of the medial retinaculum, and tenodesis to adductor magnus tendon is applied by a moderate medial force with the knee flexed at 30°. Aftercare includes immobilization of the joint limited to 30° flexion using an above-knee splint for 2 weeks. No recurrent dislocation was observed in three patients (4 knees) at a mean follow-up time of 17.7 months. Both range of motion and radiological finding were restored to normal limits.


Transcriptome analysis of Globodera pallida from the susceptible host Solanum tuberosum or the resistant plant Solanum sisymbriifolium.
A transcriptome analysis of G. pallida juveniles collected from S. tuberosum or S. sisymbriifolium 24 h post infestation was performed to provide insights into the parasitic process of this nematode. A total of 41 G. pallida genes were found to be significantly differentially expressed when parasitizing the two plant species. Among this set, 12 were overexpressed when G. pallida was parasitizing S. tuberosum and 29 were overexpressed when parasitizing S. sisymbriifolium. Out of the 12 genes, three code for secretory proteins; one is homologous to effector gene Rbp-4, the second is an uncharacterized protein with a signal peptide sequence, and the third is an ortholog of a Globodera rostochiensis effector belonging to the 1106 effector family. Other overexpressed genes from G. pallida when parasitizing S. tuberosum were either unknown, associated with a stress or defense response, or associated with sex differentiation. Effector genes namely Eng-1, Cathepsin S-like cysteine protease, cellulase, and two unknown genes with secretory characteristics were over expressed when G. pallida was parasitizing S. sisymbriifolium relative to expression from S. tuberosum. Our findings provide insight into gene regulation of G. pallida while infecting either the trap crop S. sisymbriifolium or the susceptible host, S. tuberosum.


General surgery in evolution: technology and competence.
This paper addresses the current status of General Surgery, the direction in which it is moving, and how to prepare for future changes. The paper points out that, since the predicted effects of new technology on the practice of General Surgery are more technical complexity, decreasing invasiveness, and fewer traditional operations, the consequences for the training of surgeons, the division of responsibilities among specialists, and the organization of surgical care will be substantial. In light of the current difficulty in dealing with the stresses within General Surgery, the paper discusses how changes in residency training and an increased use of Fellowships will enable the specialty to handle such disruptive changes in the future.


Indeterminate cell histiocytosis in a pediatric patient: successful treatment with thalidomide.
The 15-year-old male patient presented several 2-6 mm large livid reddish-yellowish, shiny, compact papules on the head, trunk and extremities, which had developed within the last 4 months. Histology showed normal epidermis with dense dermal infiltrate of histiocytes accompanied by few eosinophils, Touton or foamy giant cells. The histiocytes were S100 positive, CD1a negative and did not contain Birbeck granules ultrastructurally. Chest X ray, EEG, skull MRI did not show pathology. Opthalmology, neurology, oto-rhino-laryngology did not reveal alterations. Based upon the clinical symptoms and the histopathology, the diagnosis of indeterminate cell histiocytosis was confirmed. Cryotherapy and cauterization did not stop the progression of the disease, however, under thalidomide treatment no new symptoms developed and the lesions healed with pigmentation.


On the nature of keloid and hypertrophic scars.
Clinical observations make it possible in most cases to differentiate between keloid and hypertrophic scars. This suggests that different biological factors are involved in the two groups. In clinical practice in the United Kingdom most keloid scars occur spontaneously. In black-skinned people and orientals the factors of keloid and hypertrophic scars may overlap and potentiate each other.


Class I hydrophobin fusion with cellulose binding domain for its soluble expression and facile purification.
Hydrophobins, highly surface-active proteins, have the ability to reverse surface hydrophobicity through self-assembly at the hydrophilic-hydrophobic interfaces. Their unique structure and interfacial activity lead hydrophobins to have potential applications on surface functional modifications. However, class I hydrophobins are prone to self-assemble into highly insoluble amyloid-like rodlets structure. Recombinant hydrophobins could be produced by Escherichia coli but generally as an insoluble inclusion body. To overcome this insoluble expression limitation, cellulose-binding domain (CBD) from Clostridium thermocellum was fused to the N-terminal of class I hydrophobin HGFI to enhance its soluble expression in E. coli. Approximately, 94% of expressed CBD fused HGFI (CBD-HGFI) was found as soluble protein. The fused CBD could also bind specifically onto bacterial cellulose (BC) nanofibrils produced by Komagataeibacter xylinus to facilitate rapid isolation and purification of HGFI from crude extract. Lysostaphin (Lst), known as GlyGly endopeptidase could successfully cleave the flexible linker (GGGGS)2 between CBD and HGFI to recover HGFI from BC-bound CBD-HGFI. CBD-HGFI purified by immobilized metal-chelated affinity chromatography (IMAC) and Lst cleaved BC-CBD-HGFI still retained interfacial activity of hydrophobin and its effect on accelerating PETase hydrolysis against poly(ethylene terephthalate) (PET) fiber.


Differential secretion of chicken growth hormone variants after growth hormone-releasing hormone stimulation in vitro.
Variants of growth hormone (GH) are present in most vertebrates. Chicken GH (cGH) undergoes posttranslational modifications that contribute to its structural diversity. Although the 22-kDa form of GH is the most abundant, some other variants have discrete bioactivities that may not be shared by others. The proportion of cGH variants changes during ontogeny, suggesting that they are regulated differentially. The effect of growth hormone-releasing hormone (GHRH) on the release of cGH variants was studied in both pituitary gland and primary cell cultures, employing sodium dodecyl sulfate polyacrylamide gel electrophoresis, Western blotting, and densitometry. GHRH (2 nM, 2 h) stimulated the secretion of most of the size variants of cGH although the amplitude of increase was not equal for each one. A differential effect on the secretion of GH size variants, particularly on the 22- (monomer) and 26-kDa (putatively glycosylated) cGH isoforms was found in both systems. In the whole pituitary culture, the proportion of the 26-kDa immunoreactive cGH increased 35% while the 22 kDa decreased 31% after GHRH treatment in comparison with the controls. In the primary cell culture system, the proportion of the glycosylated variant increased 43% whereas the monomer and the dimer decreased 22.26 and 29%, respectively, after GHRH stimulation. Activators of intracellular signals such as 1 mM 8-bromo-cAMP and 1 microM phorbol myristate acetate had a similar effect to that obtained with GHRH. The data support the hypothesis that GH variants may be under differential control and that GHRH promotes the release of a glycosylated cGH variant that has an extended half-life in circulation.


Effect of corneal collagen crosslinking on femtosecond laser channel creation for intrastromal corneal ring segment implantation in keratoconus.
To evaluate the effect of collagen crosslinking (CXL) on femtosecond laser channel creation for intrastromal corneal ring segments (ICRS) in keratoconic eyes. Magrabi Eye Hospital, Cairo, Egypt. Comparative case series. Eyes with grade II or III keratoconus were treated by CXL. After 6 months, channel creation was performed using an IntraLase FS-60 femtosecond laser. The eyes were randomly divided into 3 groups. The default femtosecond machine power setting was 1.5 mJ in Group 1, 1.6 mJ in Group 2, and 1.7 mJ in Group 3. A control group included virgin noncrosslinked keratoconic eyes in which the default power setting was 1.5 mJ. The degree of difficulty of ICRS insertion was judged subjectively. The degree of postoperative corneal haze was recorded. Fifteen eyes of 11 patients had CXL. Each group, including the control, comprised 5 eyes. After CXL, intracorneal channel creation using the 1.5 mJ default femtosecond power setting was incomplete and mechanical dissection was required to complete the channel. When the power setting was increased to 1.6 mJ or 1.7 mJ, channel creation could be completed; however, this increased the corneal reaction (haze) postoperatively. The corneal haze resolved in all eyes within 6 weeks, and there were no further complications. Femtosecond laser channel creation can be performed after CXL; however, the laser power must be modified. Results show channel dissection and ICRS implantation should be performed before or concurrent with CXL. The author has no financial or proprietary interest in any material or method mentioned.


A Boronic Acid Assay for the Detection of Mucin-1 Glycoprotein from Cancer Cells.
Cell surface glycoproteins are commonly aberrant in disease and act as biomarkers that facilitate diagnostics. Mucin-1 (MUC1) is a prominent example, exhibiting truncated glycosylation in cancer. We present herein a boronic acid microplate assay for sensitive and high-throughput detection of such glycoproteins. The immobilization of biotin-boronic acid 1 onto streptavidin plates generated a multivalent surface for glycoprotein recruitment and detection. We first validated the binding properties of 1 in solution through titrations with alizarin dye. Next, the microplate assay was explored through horseradish peroxidase (HRP) analysis as a proof-of-concept glycoprotein with chemiluminescence detection. Finally, this platform was applied for the detection of MUC1 directly from MCF-7 human breast cancer cell lysates by using an HRP-tagged antibody that targets the cancerous form of this glycoprotein. Sensitive, dose-dependent detection of MUC1 was observed, showcasing the efficacy of this platform for detecting disease-associated glycoproteins.


Cessation of autonomous proliferation of mouse lymphoma EL4 by fusion with a T cell line.
Benzanthracene-induced C57BL/6 (H-2b) mouse T-cell lymphoma EL4 (a thymidine kinase-deficient cell line) was fused by using polyethylene glycol with an Mlsa (Mls for minor lymphocyte stimulatory) antigen-dependent T cell line, which was designated G4 and had been derived from a C3H/He mouse (H-2k), and the fused cells were cultured in HAT medium. Although no growing cells appeared in most of these fusions, we consistently obtained growth-arrested H-2Kb-positive cells from the fused cell populations by the panning method. The cells were tetraploid and were able to proliferate in response to Mlsa antigen. Three H-2Kb-positive clones, isolated by limiting dilution from three different fusions, were shown to be EL4 x G4 hybrids, because (1) they had both H-2k and H-2b antigens; (2) each of the clones had one submetacentric chromosome which was a marker chromosome of EL4, and they were tetraploid with modal chromosome numbers of 74, 78, and 79, respectively; (3) they had 4 isozymes of both parental cells. These results indicate that EL4 lymphoma cells cease to proliferate when fused with T cell line G4. The malignant phenotype of lymphoma EL4 is thus suppressed at the level of cell transformation by the introduction of the G4 cell genome.


Colour Variation in the Crocodile Lizard (Shinisaurus crocodilurus) and Its Relationship to Individual Quality.
Colour plays a key role in animal social communication including as an indicator of individual quality. Using spectrophotometry, we examined colour variation in the throat and venter of the crocodile lizard (Shinisaurus crocodilurus), an endangered species native to southern China and northern Vietnam. We detected two broad colour variants, individuals with and without red, for each body region and each sex. A cluster analysis of spectral colour measurements (hue, chroma, luminance) revealed discrete throat and ventral morphs when measured in a single snapshot in time. However, photographic evidence revealed that the amount of red relative to body size increased as they got older. Individuals with red were equally likely to be male or female and throat colour was unrelated to ventral colour. Therefore, it is premature to claim that crocodile lizards have discrete colour morphs. We used visual modelling to show that the throat and venter were easily discriminable to a lizard visual system, suggesting they function in social communication. We also asked whether colour variation signalled individual quality. Females with red throats had greater bite force while males with red throats were older. In addition, females with red venters had larger heads. We also detected differences in morphology linked to colour. Females with red throats had slender bodies and longer tails, while individuals lacking red on their throats were stouter and had shorter tails. Finally, throat and ventral colour were unrelated to reproductive output (litter size and mass) in females. Males with greater ventral luminance contrast sired offspring from litters with greater litter mass (including stillborns), while males with greater ventral chromatic contrast sired offspring whose collective live mass (excluding stillborns) was greater. Males with greater luminance contrast also sired more live offspring (excluding stillborns). Collectively, these results suggest that male ventral colour signals individual quality in males. Conservation initiatives should take colour variation into account when planning future captive breeding and release programs for this endangered species.


[The Health Examination Survey at regional level: the Emilia-Romagna Region (Northern Italy) example].
to assess time trend of lifestyles, cardiovascular risk factors, and prevalence of high-risk conditions in random samples of the general adult population residing in Emilia-Romagna, examined in two cross-sectional surveys conducted within the Epidemiological Cardiovascular Observatory (OEC 1998-2002) and the Epidemiological Cardiovascular Observatory/Health Examination Survey (OEC/HES 2008-2012). cross-sectional surveys conducted on random samples of general adult population stratified by gender and age group in some municipalities of Emilia-Romagna. in Emilia-Romagna, 341 males and 354 females were examined in 1998-2002 and 307 males and 300 females were examined in 2008-2012. cardiovascular risk factors, high risk conditions, and proportion of population following healthy lifestyles recommendations. in the second survey, prevalence of hypercholesterolemia and mean values of total and LDL cholesterol were higher. Less than 30% of the sample consumes adequate quantities of vegetables and fish; cheeses, sausages, and sweets are overconsumed, as a higher cholesterol intake. increased in total and LDL cholesterol are partly due to the reach diet, as suggested by nutrient consumption. Appropriate preventive actions for the improvement of the lifestyles and lipid profile in the general population are recommended.


Two novel laboratory tests facilitating diagnosis of glycine encephalopathy (nonketotic hyperglycinemia).
Glycine encephalopathy (GE), also known as non-ketotic hyperglycinemia, is a life-threatening metabolic disease caused by inherited deficiency of the glycine cleavage system (GCS). GE is characterized by accumulation of a large amount of glycine in serum and cerebrospinal fluids. In typical cases with GE, coma, profound hypotonia, and intractable seizures develop within several days of life. Patients with atypical symptoms may have delayed or missed diagnosis because of non-specific symptoms. It is sometimes problematic to confirm the diagnosis of GE since it requires either invasive liver biopsy for measurement of GCS activity or exhaustive mutational screening of three GCS genes, GLDC, AMT, and GCSH. We herein describe two novel laboratory tests for diagnosis of GE, [1-(13)C]glycine breath test and the multiplex ligation-dependent probe amplification (MLPA) for detection of large deletions in GLDC. The [1-(13)C]glycine breath test has been developed for noninvasive enzymatic diagnosis of GE. Because the GCS generates CO(2) by degradation of glycine, the GCS activity could be evaluated in vivo by measurement of exhaled (13)CO(2) after administration of a stable isotope, [1-(13)C]glycine. The MLPA has been developed for improvement in mutation detection rate in GE: Deletions involving multiple GDLC exons are prevalent among GE patients, but cannot be detected by the exon-sequencing analysis. Two novel diagnosis methods would facilitate diagnosis of hyperglycinemic patients as having GE.


[Uterin sarcoma, high-grade stroma, indifferenciated, referential].
High-grade endometrial stromal sarcoma (HGESS) and uterine undifferentiated sarcoma (UUS) are rare uterine malignancies arising from mesenchymal endometrial cells. They are characterized by aggressive behavior and poor prognosis. Median age of diagnostic is 55years. The most common symptoms are vaginal bleeding, abdominal pain, and pelvic mass. Approximately 65 % are diagnosed witch advance disease stage III or IV according to the International Federation of Gynecology and Obstetrics classification. Median overall survival is around 20months. The management of the disease must be discussed in multidisciplinary staff meetings. The standard management of HGESS and UUS is total hysterectomy with bilateral oophorectomy. Systematic lymphadenectomy is not recommended. Adjuvant therapies, such as chemotherapy and radiotherapy must be discussed. In case of oligo-metastasic disease, surgery of the primary tumor and metastasis must be discussed and if not operable the standard management is doxorubine-based chemotherapy.


Single-incision laparoscopic right hemicolectomy: safety and feasibility in a series of consecutive cases.
Single-incision laparoscopic colectomy (SILC) is rapidly emerging in the field of minimally invasive colon and rectal surgery. This report presents the safety and feasibility of performing SILC right hemicolectomy with the SILS™ Port Multiple Instrument Access Port. Between July 2009 and October 2009, SILC right hemicolectomy was performed for 13 consecutive, unselected patients presenting with benign or malignant pathology. Demographic data, intraoperative parameters, and postoperative outcomes were assessed. For 11 of the 13 patients (84.6%), the SILC procedure was performed with a mean incision length of 3.1 ± 1.1 cm (range, 2.5-6.0 cm) and a mean operative time of 131.5 ± 36.2 min (range, 79-180 min). Two cases required conversion to hand-assisted technique. Overall, the operative time was found to be significantly longer for the patients with a body mass index (BMI) greater than 25 kg/m(2) (152.1 ± 26.6 min) compared with the patients with a BMI less than 25 kg/m(2) (93.3 ± 11.6 min) (p < 0.002). For the subset of patients with malignant disease (five adenocarcinomas and one carcinoid), the mean number of lymph nodes extracted was 26.7 ± 14.5. There were no intraoperative complications, and the overall mean hospital stay was 2.5 ± 0.7 days (range, 2-4 days). One postoperative complication occurred (7.7%). The SILC procedure is a safe and feasible method for benign and malignant diseases requiring a right hemicolectomy. This method results in a low complication rate and a short postoperative hospital stay.


Developmentally-faithful and effective human erythropoiesis in immunodeficient and Kit mutant mice.
Immunodeficient mouse models have been valuable for studies of human hematopoiesis, but high-fidelity recapitulation of erythropoiesis in most xenograft recipients remains elusive. Recently developed immunodeficient and Kit mutant mice, however, have provided a suitable background to achieve higher-level human erythropoiesis after long-term hematopoietic engraftment. While there has been some characterization of human erythropoiesis in these models, a comprehensive analysis from various human developmental stages has not yet been reported. Here, we have utilized cell surface phenotypes, morphologic analyses, and molecular studies to fully characterize human erythropoiesis from multiple developmental stages in immunodeficient and Kit mutant mouse models following long-term hematopoietic stem and progenitor cell engraftment. We show that human erythropoiesis in such models demonstrates complete maturation and enucleation, as well as developmentally appropriate globin gene expression. These results provide a framework for future studies to utilize this model system for interrogating disorders affecting human erythropoiesis and for developing improved therapeutic approaches.


Deprivation: different implications for forensic psychiatric need in urban and rural areas.
Ecological relationships between deprivation and forensic psychiatric admission rates may differ in urban and rural areas. The aim of the study was to compare the relationship between material deprivation and forensic admission rates in rural and urban areas for a whole-national service in Ireland over a 3-year period. All Irish forensic admissions from 1997 to 1999 were allocated to the appropriate small area. Material deprivation scores were calculated from census data. Mean annual admission rates and Bayesian standardised forensic admission ratios for small areas were aggregated by material deprivation score and population density. At small area level, there were significant non-linear increases in forensic admissions with increasing deprivation. The increases in urban areas (population density >10/hectare) were absent in less densely populated areas. Deprivation alone may not be the key factor in predicting forensic service utilisation. Factors associated with specifically urban deprived areas may be of greater relevance in planning services.


Hirschsprung-associated inflammatory bowel disease: A multicenter study from the APSA Hirschsprung disease interest group.
A small number of Hirschsprung disease (HD) patients develop inflammatory bowel disease (IBD)-like symptoms after pullthrough surgery. The etiology and pathophysiology of Hirschsprung-associated IBD (HD-IBD) remains unknown. This study aims to further characterize HD-IBD, to identify potential risk factors and to evaluate response to treatment in a large group of patients. Retrospective study of patients diagnosed with IBD after pullthrough surgery between 2000 and 2021 at 17 institutions. Data regarding clinical presentation and course of HD and IBD were reviewed. Effectiveness of medical therapy for IBD was recorded using a Likert scale. There were 55 patients (78% male). 50% (n = 28) had long segment disease. Hirschsprung-associated enterocolitis (HAEC) was reported in 68% (n = 36). Ten patients (18%) had Trisomy 21. IBD was diagnosed after age 5 in 63% (n = 34). IBD presentation consisted of colonic or small bowel inflammation resembling IBD in 69% (n = 38), unexplained or persistent fistula in 18% (n = 10) and unexplained HAEC >5 years old or unresponsive to standard treatment in 13% (n = 7). Biological agents were the most effective (80%) medications. A third of patients required a surgical procedure for IBD. More than half of the patients were diagnosed with HD-IBD after 5 years old. Long segment disease, HAEC after pull through operation and trisomy 21 may represent risk factors for this condition. Investigation for possible IBD should be considered in children with unexplained fistulae, HAEC beyond the age of 5 or unresponsive to standard therapy, and symptoms suggestive of IBD. Biological agents were the most effective medical treatment. Level 4.


Leukocyte count predicts microembolic Doppler signals during carotid stenting: a link between inflammation and embolization.
Protected stenting has emerged as a safe and effective alternative to endarterectomy for the treatment of carotid stenosis in patients at high operative risk. Distal microembolization occurs invariably during carotid stenting. Little is known about the relationship between systemic inflammation and embolization during carotid stenting. We examined 43 consecutive patients who underwent carotid stenting with simultaneous transcranial Doppler (TCD) monitoring of the ipsilateral middle cerebral artery. Embolization was quantified by measuring microembolic signals (MES) on TCD. Preprocedure leukocyte counts were related to MES. In unadjusted analyses, preprocedure leukocyte count was positively correlated with total procedural MES (r2= 0.16; P=0.008). After considering age, gender, comorbidities, concomitant medical therapies, and the use of emboli prevention devices, increasing leukocyte count (beta=35 for each 1000/microL increment; P=0.018) remained a significant and independent predictor of embolization (model-adjusted r2=0.365; P=0.0005). Increasing preprocedure leukocyte count independently predicted more frequent MES during carotid stenting. These data suggest that systemic inflammation may influence the degree of procedural embolization.


Study of T helper 1 and T helper 2 responses in pemphigus vulgaris patients receiving interferon alpha 2a injections in addition to a standard protocol therapy: a randomized controlled trial.
T helper (Th)1 insufficiency was recently found to be related to the pathogenesis of pemphigus vulgaris (PV). Decreased Th1 response was particularly noticed in the early stages of PV. Therefore, administration of interferon alpha in the early stages of aggressive PV may lead to rapid control of the acute stage of the disease. Our aim was to evaluate the role of interferon alpha in the treatment of PV. 30 patients with acute severe PV (>60 % affection) and 30 age and sex-matched healthy subjects were included in this RCT. Patients were randomly divided into two groups (A and B). Group B patients received interferon retard (one subcutaneous injection/week for 4 weeks) in addition to our protocol for the treatment of PV (systemic pulse corticosteroids/cyclophosphamide in combination with sulphasalazine and pentoxifylline) that was administered to all the included patients. IFN-γ and IL-4 were estimated by ELISA before treatment, after 4 weeks and at the end of the study duration (12 weeks). Clinical assessment was done by PAAS on a biweekly basis. All PV patients showed significantly (P < 0.001) elevated levels of IL-4 and significantly (P < 0.001) depressed mean concentration of IFN-γ as compared with healthy controls. Twelve weeks after therapy both groups showed significant improvement in their mean PAAS being more evident and more rapid in group B. IFN-γ was elevated significantly and IL-4 was dropped significantly in group B patients in comparison to group A (P < 0.001). As a conclusion, interferon therapy in severe PV could achieve a more prompt and better clinical response.


Radioimmunological identification of tropoelastin.
Antiserum was prepared in sheep against insoluble elastin isolated from embryonic-chick aortae. In an indirect immunoprecipitation test, the antiserum reacted quantitatively with small amounts of radioactively labelled purified tropoelastin prepared from embryonic-chick aortae. The antiserum did not cross-react with chick procollagen, and the antiserum uas used to identify radioactively labelled tropoelastin secreted by chick aorta cells in suspension culture.


Effect of diet upon ventilation and effort perception during a physical stressor.
The present research explored the effects of a high and low carbohydrate diet on ratings of perceived exertion (RPE) during a 10 minute bicycle ergometer test at high physical work capacity (70% of maximum aerobic capacity). Twelve female student volunteers participated and were randomly assigned the order of diet treatments in which they served as their own control and/or experimental group. Following each dietary regimen, subjects were instructed to pedal the bicycle ergometer for ten minutes at an intensity of 70% of their maximum capability. Throughout the cycling protocol, RPE scores were recorded at 2, 4, 6, 8, and 10 minutes; in addition expiratory gases were collected and analyzed by a computerized indirect calorimetry system. Results in the current investigation indicated that an increased carbohydrate intake resulted in an "ergogenic" or "work-producing" effect by attenuating the metabolic acidosis, possibly diminishing the sensations of the central cardiopulmonary effort, further attenuating the psychophysiological perceptions of effort. Conversely, ingestion of a low carbohydrate diet resulted in accentuation of the ventilatory drive eliciting higher signals of exertion during dynamic exercise.


Reproducibility analysis of the computerized tomography angiography-derived left atrial wall thickness maps.
Adapting the ablation index (AI) to the left atrial wall thickness (LAWT) derived from computed tomography angiography (CTA) allows for a personalized approach that showed to improve PVI safety and outcomes. Three observers with different degrees of experience performed complete LAWT analysis of CTA for 30 patients and repeated the analysis for 10 of these patients. Intra- and inter-observer reproducibility of these segmentations was assessed. Geometric congruence of repeated reconstruction of LA endocardial surface showed that 99.4% of points in the 3D reconstructed mesh were within < 1 mm distance for the intra-observer variability and 95.1% for the inter-observer. For the LA epicardial surface, an 82.4% of points were within < 1 mm for intra-observer and a 77.7% for inter-observer. A 1.99% of points were further than 2 mm for the intra-observer and a 4.1% for the inter-observer. Colour agreement between LAWT maps showed that a 95.5% and a 92.9% intra- and inter-observer respectively presented the same colour or a change to the colour immediately above or below. The ablation index (AI), which was adapted to this LAWT colour maps to perform a personalized pulmonary vein isolation (PVI), showed an average difference in the derived AI lower than 25 units in all cases. For all analyses, the concordance increased with user-experience. Geometric congruence of LA shape was high, for both endocardial and epicardial segmentations. LAWT measurements were reproducible, increasing with user experience. This translated into a negligible impact in the target AI.


Endoscopic submucosal dissection in large sessile lesions of the rectosigmoid: learning curve in a European center.
Endoscopic submucosal dissection (ESD) in the colorectum is not currently a standard procedure. Few data are available from the Western world. The aim of the present study was to report on the first experiences and the learning curve of colorectal ESD in a European center. A total of 82 rectosigmoid lesions were referred for ESD. Lesion characteristics, resection rates, procedure times, complications, and recurrences were recorded prospectively. Results were compared between three consecutive study periods in order to determine the learning curve. Lesions were located in the rectum (86.6 %) and the sigmoid colon (13.4 %). Median diameter was 45.5 mm. Lesions were of Paris type 0-Is with pit pattern type V (n = 8), 0-IIa (n = 33), 0-IIa + Is (n = 36), and 0-IIa + IIc (n = 5). The malignancy rate in these groups was 100 %, 0 %, 14 %, and 20 %, respectively. ESD was possible in 76 lesions (92.7 %). En bloc resection rate and R0 resection rate were 81.6 % and 69.7 %, respectively. Median procedure time was 176 minutes. Between the three consecutive study periods, en bloc resection rate increased (60.0 %, 88.0 %, 96.2 %), R0 resection rate increased (48.0 %, 76.0 %, 84.5 %; P < 0.001), and procedure time decreased (200, 193, 136 minutes; P = 0.027). The perforation and bleeding rates were 1.3 % and 7.9 %, respectively. Recurrence risk was 0 % after R0 en bloc resection and 41.7 % after piecemeal resection (P < 0.05). Median follow-up was 23.6 months. In the European setting, ESD in the distal colon is feasible with acceptable complication risks. Resection rates were not as high as those from Japanese studies; however, a clear learning curve could be shown. Colorectal ESD needs to be further evaluated, particularly in Europe where ESD experience is low.


Dental education in primary care: 14 years of Peninsula Dental School.
Peninsula Dental School, established in 2006, was the UK's first new dental school in 40 years. It had the freedom to develop a completely new dental education curriculum planned on pedagogic thinking, designed to equip the dental care professionals of the twenty-first century. This was based on three distinct pillars: professionalism (developing a student's trust in their own autonomy); dental skills of the highest order (not just technical skills but also communication skills); and social engagement. As such, a truly innovative approach to dental education was created that has strong roots in evidence.This paper describes the University of Plymouth Peninsula Dental School's achievements against these initial objectives under the following areas: training in primary care; a novel spiralling integrated curriculum and assessments; facilities reaching out to deliver patient care; bringing meaningful patient contact to students from the earliest months of their course; embedding community engagement within the curriculum; development of Peninsula Dental Social Enterprise; and team working, training a variety of dental care profession students side by side.The University of Plymouth Peninsula Dental School, working with all its partners, has successfully pioneered and delivered significant changes in the field of education and continues to strive to further develop these and more for the future.


How older Vietnamese Australian women manage their medicines.
Safe use of medicines is relevant to all, but especially older people, primarily because they have ageing bodies that require more medicines and are therefore more likely to experience complications, including adverse drug interactions. Australia has a rapidly growing migrant older population composed of people with different beliefs about, and practices using, medicines. This paper presents qualitative findings from interviews and focus groups conducted with older Vietnamese-Australian women living in Sydney, Australia. The findings show how the women's health literacy influenced their medication use and explain the cultural reasons behind their decision to vary medicine use from that prescribed by their western trained doctors. Concern that health professionals do not favour combining western with traditional medicines led some participants to manage their medicines without advice from their doctor. The findings support recommendations to reduce the likelihood of adverse medication outcomes by increasing health professionals' cultural competence, encouraging patients to work with their doctor and report all medications taken, and increasing funding for research into the effects of combining western with traditional medicines.


Use of naloxone to reverse carfentanil citrate-induced hypoxemia and cardiopulmonary depression in Rocky Mountain wapiti (Cervus elaphus nelsoni).
With the use of a crossover study design, we investigated the respiratory and cardiovascular effects of naloxone administration in eight healthy Rocky Mountain wapiti (Cervus elaphus nelsoni) anesthetized with carfentanil (10 microg/kg i.m.) and xylazine (0.1 mg/kg). Anesthetized animals showed profound hypoxemia with mild hypercapnia, tachycardia, hypertension, and acidosis prior to naloxone administration. After monitoring equipment was placed, animals were administered either naloxone (2 microg/microg carfentanil i.v.) or an equivalent volume of normal saline. Mean values for PaO2, PaCO2, heart rate, and respiratory rate were significantly different between naloxone- and saline-treated groups, but mean blood pressure, hematocrit, and serum electrolyte concentrations were not. Mean PaO2 was 23.0 +/- 4.1 mm Hg prior to administration of naloxone or saline and increased to 50.2 +/- 7.3 mm Hg after naloxone administration. Mean PaO2 of saline-treated animals did not change significantly. Electrocardiograms of three saline-treated animals suggested myocardial hypoxia. Hypoxemia appeared to be caused by respiratory depression, hemodynamic alterations, and lateral recumbency. All but one animal remained anesthetized after naloxone administration. Anesthesia in all animals was reversed in < or = 4 min with naltrexone (100 mg/mg carfentanil i.v. s.c.) and yohimbine (0.1 mg/kg i.v.). One bolus of naloxone improved oxygenation in carfentanil-xylazine-anesthetized wapiti.


Chest pain without established ischaemic heart disease in primary care patients: associated comorbidities and mortality.
Ischaemic heart disease (IHD) can be excluded in the majority of patients with unspecific chest pain. The remainder have what is generally referred to as non-cardiac chest pain, which has been associated with gastrointestinal, neuromusculoskeletal, pulmonary, and psychiatric causes. To assess morbidity and mortality following a new diagnosis of non-specific chest pain in patients without established IHD. Population-based cohort study with nested case-control analysis. UK primary care practices contributing to the General Practice Research Database. Patients aged 20-79 years with chest pain who had had no chest pain consultation before 2000 and no IHD diagnosis before 2000 or within 2 weeks after the index date were selected from the General Practice Research Database. The selected 3028 patients and matched controls were followed-up for 1 year. The incidence of chest pain in patients without established IHD was 12.7 per 1000 person-years. In the year following the index date, patients who had chest pain but did not have established IHD were more likely than controls to receive a first IHD diagnosis (hazard ratio [HR] = 18.2, 95% confidence interval [CI] = 11.6 to 28.6) or to die (HR = 2.3, 95% CI = 1.3 to 4.1). Patients with chest pain commonly had a history of gastro-oesophageal reflux disease (GORD; odds ratio [OR] = 2.0, 95% CI = 1.5 to 2.7) or went on to be diagnosed with GORD (risk ratio 4.5, 95% CI = 3.1 to 6.4). Patients with chest pain but without established IHD were found to have an increased risk of being diagnosed with IHD. Chest pain in patients without established IHD was also commonly associated with GORD.


Reducing Outpatient Antibiotic Prescribing for Acute Respiratory Infections: A Quasi-Experimental Study.
Approximately 50% of antibiotics prescribed are not necessary; nevertheless, in the United States among the many outpatient prescriptions, there are few more widely prescribed than antibiotics. The overuse of antibiotics to treat viral infections has been largely responsible for the emergence of antibiotic resistance. A quasi-experimental study was conducted among a sample of eight urgent care providers who received the 1-hour Reducing Outpatient Antibiotic Resistance (ROAR) educational intervention on antibiotic overuse and appropriate prescribing. Outcomes measured included provider antibiotic prescribing rates for viral illnesses before and after intervention, prescribing practices among the providers, and provider awareness and beliefs regarding antibiotic prescribing and resistance. The antibiotic prescribing rate decreased from 30% to 20% after intervention, p = .078. Pre-intervention patients had a 3.3 times (p = .001) and post-intervention patients had a 4.2 times (p ≤ .0005) greater likelihood of being prescribed an antibiotic if they were seen by a physician than if they were seen by a nurse practitioner. Within their setting, 87% believed antibiotics are overused, and 99% believed antibiotic resistance is a problem. Additional research utilizing the ROAR intervention is necessary to evaluate its effect on antibiotic prescribing in the urgent care setting.


Interpretation of In Vitro Concentration-Response Data for Risk Assessment and Regulatory Decision-making: Report from the 2022 IWGT Quantitative Analysis Expert Working Group Meeting.
Quantitative risk assessments of chemicals are routinely performed using in vivo data from rodents; however, there is growing recognition that non-animal approaches can be human-relevant alternatives. There is an urgent need to build confidence in non-animal alternatives given the international support to reduce the use of animals in toxicity testing where possible. In order for scientists and risk assessors to prepare for this paradigm shift in toxicity assessment, standardization and consensus on in vitro testing strategies and data interpretation will need to be established. To address this issue, an Expert Working Group (EWG) of the 8th International Workshop on Genotoxicity Testing (IWGT) evaluated the utility of quantitative in vitro genotoxicity concentration-response data for risk assessment. The EWG first evaluated available in vitro methodologies and then examined the variability and maximal response of in vitro tests to estimate biologically relevant values for the critical effect sizes considered adverse or unacceptable. Next, the EWG reviewed the approaches and computational models employed to provide human-relevant dose context to in vitro data. Lastly, the EWG evaluated risk assessment applications for which in vitro data are ready for use and applications where further work is required. The EWG concluded that in vitro genotoxicity concentration-response data can be interpreted in a risk assessment context. However, prior to routine use in regulatory settings, further research will be required to address the remaining uncertainties and limitations. This article is protected by copyright. All rights reserved.


Elevated sTREM2 and NFL levels in patients with sepsis associated encephalopathy.
Sepsis-associated encephalopathy (SAE) is a common manifestation of sepsis that may lead to cognitive decline. Our aim was to investigate whether the neurofilament light chain (NFL) and soluble triggering receptor expressed on myeloid cells 2 (sTREM2) could be utilized as prognostic biomarkers in SAE. In this prospective observational study, baseline serum levels of sTREM2 and cerebrospinal fluid (CSF) levels of sTREM2 and NFL were measured by ELISA in 11 SAE patients and controls. Patients underwent daily neurological examination. Brain magnetic resonance imaging (MRI) and standard electroencephalography (EEG) were performed. Cognitive dysfunction was longitudinally assessed after discharge in 4 SAE patients using the Mini-Mental State Examination (MMSE) and Addenbrooke's Cognitive Examination-Revised (ACE-R) tests. SAE patients showed higher CSF sTREM2 and NFL levels than controls. sTREM2 and NFL levels were not correlated with the severity measures of sepsis. Three months after discharge, 2 SAE patients displayed ACE-R scores congruent with mild cognitive impairment (MCI), persisting in one patient 12 months after discharge. SAE patients with MCI showed higher CSF NFL levels, bacteremia, and abnormal brain MRI. Patients with increased serum/CSF sTREM2 levels showed trends towards displaying poorer attention/orientation and visuo-spatial skills. sTREM2 and NFL levels may serve as a prognostic biomarker for cognitive decline in SAE. These results lend further support for the involvement of glial activation and neuroaxonal degeneration in the physiopathology of SAE.


[Robot-assisted surgery as an elective-fascinating lesson(s)?]
Even though robot-assisted operations have evolved to a standard procedure in surgery, they are underrepresented in the curriculum of current medical students. We present our experience and findings in Germany's first elective "Robot-assisted surgery" at a urological department for undergraduate medical students. Ten undergraduates in their final years were taught the theoretical basics and practical skills in robot-assisted surgery within six lessons each lasting 2 h, including the opportunity to observe a live robot-assisted surgery. The increase of knowledge (ten multiple-choice questions) and skills (exercises Camera 0, Clutch, and Sea Spikes 1) on a robotic simulation device were quantified including an evaluation of the student's perspective. The 10 participants had a significant increase in knowledge and gave at a median of 3.5 additional correct answers in the final assessment (p = 0.011). For two out of three practical exercises, the overall score significantly increased (Camera 0 and Sea Spikes 1, for both p < 0.05), but for the exercise "Clutch", only economy of motion significantly improved (p = 0.028). The elective was evaluated (very) good and the willingness of the participants to become urologists significantly increased (p = 0.007). There is a great interest of many undergraduate medical students in robot-assisted surgery. Offering an elective appears to be an excellent format to teach the theoretical background and practical skills in robotic (urologic) surgery. Moreover, such an elective could raise more attention to the field of urology and might attract future colleagues.


The 50 most influential original articles in vascular surgery during the last 25 years.
We have compiled a list of the 50 most-cited original articles in the field of vascular surgery during the last 25 years to highlight the important changes in practice that have occurred during this interval and provide surgical trainees in vascular surgery ready access to such influential articles. A Web of Knowledge Citation Index Search was performed in December 2013 for the most-cited journal articles in the discipline of vascular surgery. We searched the term "vascular" in the cited reference search area and then further narrowed our results to exclude all categories except "surgery," "general internal medicine," and "cardiac/cardiovascular systems." We included only documents labeled as "articles" and those published in English. Articles dealing with cardiac surgery, interventional cardiology, and cardiovascular biology were excluded. Our search period was from January 1, 1988, through December 3, 2013. The 50 most frequently cited works were chosen, and a citation density was calculated for each, reflecting the average number of citations each received per year since publication. The articles were then sorted into a defined category, based on the clinical subject to which they pertained. The Citation Index Search resulted 80,379 articles, of which the top 50 were indexed and organized according to their citation density and area within the scope of clinical vascular surgery. The number of citations ranged from 218 to 3593. The median citation density was 50.2 (range, 11.3-201.3). This report is a representation of the most-cited original publications in the field of clinical vascular surgery during the last 25 years. This is an effort to highlight the seminal works that have shaped the discipline of vascular surgery as well as to provide a concise reference list for the surgical trainee in the process of his or her education.


Reversible inhibition of electron transfer in the ubiquinol. Cytochrome c reductase segment of the mitochondrial respiratory chain in hibernating ground squirrels.
Electron transfer through the ubiquinol:cytochrome c1-segment of liver mitochondria isolated from hibernating ground squirrels Citellus undulatus is repressed by 70-80% as compared to mitochondria from the active animals. The inhibition site is likely to be localized between ubiquinone and the cytochrome bc1 complex. Partial release of the inhibition can be observed upon swelling of the isolated mitochondria in a hypoosmotic medium, the effect being prevented by phospholipase A2 inhibitors. Possible role of phospholipase A2 in regulation of ubiquinol oxidation by complex bc1 is discussed.


Seasonal variation and complex analysis of microplastic distribution in different WWTP treatment stages in Lithuania.
Industrial wastewater, domestic wastewater, and stormwater are the three entry points for microplastics (MP) in wastewater treatment plants. Extreme weather conditions, such as rising temperatures and heavy rainfall caused by climate change, can alter the rate at which MP enters wastewater treatment plants. In this study, wastewater and sludge samples from different treatment stages were collected during a 12-month sampling campaign (seasonal) to determine the efficiency of a municipal wastewater treatment plant in removing microplastic particles. MP ranging from 20 to 1000 µm were detected and classified by shape, color, size, and chemical composition. All samples contained MP particles, with concentration ranging from 1964 ± 50-2982 ± 54 MP/L in influent to 744 ± 13-1244 ± 21 MP/L in effluent and 91.1 ± 8-61.9 ± 5 MP/g in sludge; 71.6-90.1% identified particles were fragment-type with black, white, and transparent colors. Most of the microplastic particles were removed in the activated sludge tank, while the average removal rate in the wastewater treatment plant was 57%. The total concentration of MP was 27% higher in spring than in other seasons. The most common microplastic particles were polyethylene terephthalate (PET), polystyrene (PS), and polypropylene (PP). These results demonstrate the value of long-term monitoring and MP quantification, which would provide a more accurate estimate of MP pollution from wastewater treatment plants.


Changes in Assessment of and Satisfaction With Discharge Preparation From the Neonatal Intensive Care Unit.
A successful transition from the neonatal intensive care unit (NICU) to home is aided by a comprehensive discharge planning program that keeps families involved and engaged with the discharge preparation process. To compare the assessment of parental NICU discharge preparedness with parental satisfaction with the NICU discharge preparation. Families were surveyed 4 to 6 weeks after NICU discharge, and those selecting "very prepared" were considered "satisfied" with their discharge preparation. On discharge day, families were considered "prepared" for discharge based on their overall level of preparedness and their nurse's rating of them on a discharge readiness assessment tool. In total, 1104 families (60%) reported being both "satisfied" and "prepared"; 293 families (16%) were "satisfied" but not "prepared"; 297 families (16%) were not "satisfied" but were "prepared"; and 134 families (7%) were neither "satisfied" nor "prepared." Compared with families that were both "satisfied" and "prepared," families that were neither "satisfied" nor "prepared" were more likely to be raising the infant alone, of Black race, and to have sicker infants. Some families are at a higher risk and merit more consideration during NICU discharge planning. Assess the discharge readiness of all families prior to discharge. Those at an increased risk may benefit from more discharge education and training, specifically for single mothers, those with limited resources, or others considered at high risk.


Roles of factor Va heavy and light chains in protein and lipid rearrangements associated with the formation of a bovine factor Va-membrane complex.
Factor Va is an essential protein cofactor of the enzyme factor Xa, which activates prothrombin to thrombin during blood coagulation. Peptides with an apparent Mr of approximately 94,000 (heavy chain; HC) and approximately 74,000 or 72,000 (light chain; LC) interact in the presence of Ca2+ to form active Va. The two forms of Va-LC differ in their carboxyl-terminal C2 domain. Using Va reconstituted with either LC form, we examined the effects of the two LC species on membrane binding and on the activity of membrane-bound Va. We found that 1) Va composed of the 72,000 LC bound only slightly more tightly to membranes composed of a mixture of neutral and acidic lipids, the Kd being reduced by a factor of approximately 3 at 5 mM and by a factor of 6 at 2 mM Ca2+. 2) The two forms of Va seemed to undergo different conformational changes when bound to a membrane. 3) The activity of bovine Va varied somewhat with LC species, the difference being greatest at limiting Xa concentration. We have also addressed the role of the two Va peptides in membrane lipid rearrangements and binding: 1) Va binding increased lateral packing density in mixed neutral/acidic lipid membranes. In the solid phase, Va-HC had no effect, whereas Va-LC and whole Va had similar but small effects. In the fluid phase, Va-HC and whole Va both altered membrane packing, with Va-HC having the largest effect. 2) Va-HC bound reversibly and in a Ca2+-independent fashion to membranes composed of neutral phospholipid (Kd, approximately 0.3 microM; stoichiometry approximately 91). High ionic strength had little effect on binding. 3) The substantial effect of Va on packing within neutral phospholipid membranes was mimicked by Va-HC. 4) Based on measurements of membrane phase behavior, binding of Va or its peptide components did not induce thermodynamically discernible lateral membrane domains. These results suggest that the membrane association of factor Va is a complex process involving both chains of Va, changes in lipid packing, and changes in protein structure.


Ulcerated Spitz nevus masquerading as a juvenile xanthogranuloma.
We report a case of ulcerated atypical Spitz nevi that demonstrated a yellow to light orange background under dermoscopy, which can be seen in juvenile xanthogranuloma (JXG) and is referred to as the "setting sun" appearance. This yellow to orange appearance was due to serous crusting and not histiocytic infiltration, which is seen in JXG. This case highlights overlapping dermatoscopic features between the two skin lesions and polymorphous vascular structures, which are unique to atypical Spitz nevi.


Photodecay dynamics of octaethylporphine in the condensed phase explored via resonance Raman spectroscopy and density functional theory calculation.
Resonance Raman spectra of free-base octaethylporphine (OEP) were obtained with 368.9 nm, 397.9 nm and 416.0 nm excitation wavelengths, and density functional calculations were done to help the elucidation of Soret (B(x) and B(y)-band) electronic transitions and the corresponding photo relaxation dynamics of OEP. The RRs indicate that the Franck-Condon region photo relaxation dynamics upon S(0)→S(8) electronic transition is predominantly along the totally symmetric C(m)C(α) stretch, the C(β)C(β) stretch, and simultaneously along the asymmetric δ(pyr deformation),γ(CH(2)) vibrational relaxation processes. The excited state structural dynamics of OEP determined from resonance Raman spectra show that the internal conversion between B(y) and B(x) electronic states occurs in tens of femtoseconds and the electronic relaxation dynamics were firstly interpreted with account of the time-dependent wave packet theory and Herzberg-Teller (vibronic coupling) contributions.


Regulatory role of ATP at developing neuromuscular junctions.
Neuronal factors co-released with neurotransmitters may play an important role in synaptic development and function. Extracellular application of adenosine 5'-triphosphate (ATP), a substance co-stored and co-released with acetylcholine (ACh) in peripheral nervous systems, potentiated the spontaneous secretion of ACh at developing neuromuscular synapses in Xenopus 1-day-old cell cultures, as shown by a marked increase in the frequency of spontaneous synaptic currents recorded in the post-synaptic muscle cell. ATP also increased the frequency of miniature endplate potentials in the isolated tails of 2-week-old Xenopus tadpoles, with much smaller effect than that observed in cell cultures. The potentiation effect of ATP on ACh release in Xenopus cell cultures was inhibited by L-type Ca2+ channel blockers, suggesting that the L-type Ca2+ channel is responsible for the positive regulation of spontaneous ACh secretion by ATP at the developing neuromuscular synapses. The frequency of spontaneous synaptic events was found to vary greatly from cell to cell in the culture, over two orders of magnitude. Synapses with high frequency events are probably under the influence of endogenously released ATP. In addition, ATP was shown to potentiate the responses of isolated myocytes to iontophoretically-applied ACh. Local application of ATP to one region of the elongated myocyte surface resulted in potentiated ACh responses only at the ATP-treated region. Single channel recording showed that ATP specifically increased the open time and opening frequency of embryonic-type, low conductance ACh channels. Pharmacological experiments suggest that ATP exerted both its pre- and post-synaptic effects by binding to P2-purinoceptors and activating protein kinase C. Moreover, the potentiation effects of ATP were restricted to the early stages of embryos. Taken together, these results suggest that ATP co-released with ACh or released from stimulated myocytes may promote synaptic development by potentiating pre-synaptic ACh release and post-synaptic ACh channel activity during the early phase of synaptogenesis.


Construction of heme enzymes: four approaches.
Construction of enzymes that catalyze either desired reactions or exhibit desired substrate specificity is one of the goals of enzymatic study. Rational design of enzymes is an important approach in this field. Another, extremely different, methodology from rational design, directed evolution, has been rapidly developed over the past two years.


Synthetic enhancer compounds, besides acting on biogenic amine system, influence the glutamate transmission and stress response.
Pharmaceutically available enhancer selegiline/(-)-deprenyl (DEP) in the clinically used dose shows antidepressant effect, but nothing is known about this effect in enhancer dose, and its effect on co-morbid anxiety. Moreover, data about the antidepressant/antianxiety effects of the serotonin-influencing enhancer, (2R)-1-(1-benzofuran-2-yl)-N-propylpentane-2-amine (BPAP) are also missing. The aim of the present paper is to establish the role of enhancer regulation in anxiety and follow the changes in the phosphorylation of glutamate subunits in prefrontal cortex as well as stress-related organ and hormonal changes as possible background mechanism. The effect of 3-week-treatment of rats with specific (0.001 mg/kg for DEP, 0.0001 mg/kg for BPAP) and non-specific (0.1 mg/kg for DEP, 0.05 mg/kg for BPAP) enhancer doses were evaluated on anxiety-like behavior in the elevated plus maze (EPM) and open-field (OF) tests. Phosphorylated glutamatergic GluR1 and GluN2B subunits were analyzed by Western blot. Changes in the stress-regulatory system were evaluated by measuring the organ weights and blood corticosterone concentrations. Non-specific enhancer doses had a tendency for anxiolysis on EPM, while only 0.1 mg/kg DEP elevated motility in OF. Specific enhancer doses significantly increased the expression of both glutamatergic receptor subunits; non-specific doses elevated only pGluR1. Treatments had no effects on stress-like organ weights; however, the specific enhancer doses significantly reduced the dark phase resting corticosterone levels. The study proved the enhancer-sensitivity of the glutamatergic transmitter system and suggested enhancer-induced stabilization of stress-hormone levels without major impact on non-stimulated anxiety-like behavior.