Identification of DNA binding proteins in vaccinia virus by DNA-protein crosslinking.
DNA binding proteins of vaccinia virus (VV) virions, strain LIVP, have been studied by their covalent crosslinking to DNA, using two-dimensional gel retardation electrophoresis of crosslinked DNA and proteins as well as the 'protein image' hybridization assay. Five proteins with molecular masses of 16, 25, 27, 41 and 54 kDa, respectively, associated with all analysed DNA sequences, including early and late genes and their promoters, have been identified.


[Dumping syndrome: Diagnostics and therapeutic options].
Dumping syndrome is a common complication after surgery of the upper gastrointestinal tract with symptoms ranging from mild gastrointestinal discomfort and moderate vasomotor disturbances, to severe hyperinsulinemic hypoglycemia. Due to the increasing number of bariatric procedures being performed worldwide, bariatric surgery has become the most common cause for this disease entity. The aim of this review is to highlight the evidence for the physiological mechanisms contributing to dumping syndrome after the two most common bariatric surgery procedures, Roux-en-Y gastric bypass and sleeve gastrectomy, to discuss technical aspects of the procedures underlying the development of the syndrome, patient-related predictive factors and other differential diagnoses, together with diagnostic and therapeutic algorithms.


Enhancement of histamine synthesis in mouse skin by epinephrine.
Intravenous administration of 10-40 micrograms/Kg epinephrine to mice leads to a transient 50% increase in skin histamine, followed by an increase in blood histamine. Delayed inhibition by alpha-fluorometyl histidine (alpha FMH), suggests that these changes follow stimulation of pre-formed tissue histidine decarboxylase.


Characterization of D-lactate dehydrogenase from Pediococcus acidilactici that converts phenylpyruvic acid into phenyllactic acid.
The gene coding for D-lactate dehydrogenase (D-LDH) from Pediococcus acidilactici DSM 20284 was cloned and expressed in E. coli. The recombinant enzyme was purified by nickel-affinity chromatography. It converted phenylpyruvic acid (PPA) to 3-phenyllactic acid maximally at 30°C and pH 5.5 with a specific activity of 140 and 422 U/mg for PPA and pyruvate, respectively. The K(m), turnover number (k(cat)), and catalytic efficiency (k(cat)/K(m)) for PPA were 2.9 mM, 305 s(-1), and 105 mM(-1) s(-1), respectively.


Patients with polyclonal hepatocellular carcinoma are at a high risk of early recurrence and have a poor recurrence-free survival period.
Tumor heterogeneity based on copy number variations is associated with the evolution of cancer and its clinical grade. Clonal composition (CC) represents the number of clones based on the distribution of B-allele frequency (BAF) obtained from a genome-wide single nucleotide polymorphism (SNP) array. A higher CC number represents a high degree of heterogeneity. We hypothesized and evaluated that the CC number in hepatocellular carcinoma (HCC) tissues might be associated with the clinical outcomes of patients. Somatic mutation, whole transcriptome, and CC number based on copy number variations of 36 frozen tissue samples of operably resected HCC tissues were analyzed by targeted deep sequencing, transcriptome analysis, and SNP array. The samples were classified into the heterogeneous tumors as poly-CC (n = 26) and the homogeneous tumors as mono-CC (n = 8). The patients with poly-CC had a higher rate of early recurrence and a significantly shorter recurrence-free survival period than the mono-CC patients (7.0 months vs. not reached, p = 0.0084). No differences in pathogenic non-synonymous mutations, such as TP53, were observed between the two groups when targeted deep sequencing was applied. A transcriptome analysis showed that cell cycle-related pathways were enriched in the poly-CC tumors, compared to the mono-CC tumors. Poly-CC HCC is highly proliferative and has a high risk of early recurrence. CC is a possible candidate biomarker for predicting the risk of early postoperative recurrence and warrants further investigation.


The genus Dryadaula Meyrick (Lepidoptera, Tineoidea, Dryadaulidae) in China, with descriptions of four new species and a world checklist.
Four new species of the genus Dryadaula Meyrick, 1893 from China are described: Dryadaulaauriformis sp. nov., D.flavostriata sp. nov., D.hirtiglobosa sp. nov. and D.securiformis sp. nov. Photographs of adults and genitalia of the new species are provided. DNA barcodes of D.auriformis sp. nov., D.hirtiglobosa sp. nov. and D.securiformis sp. nov. are given. A key to the species in China and a detailed checklist for the genus with all 49 known species in the world are presented.


Endothelin-1 increases CHSY-1 expression in aortic endothelial cells via transactivation of transforming growth factor β type I receptor induced by type B receptor endothelin-1.
TGF-β through hyperelongation of glycosaminoglycan (GAG) chains leads to binding of low-density lipoproteins to the proteoglycans. The vasoactive peptide, endothelin-1 (ET-1), plays a key role in the development of atherosclerosis. This study addressed the question whether ET-1 by activating the Rho kinase and cytoskeletal rearrangement can transactivate the TGF-β receptor leading to phosphorylation of the transcription factor Smad2 and increased expression of the GAG chain synthesizing enzyme such as chondroitin synthase-1 (CHSY-1) in bovine aortic endothelial cells (BAECs). In this study, intermediates in ET-1-induced Smad2C phosphorylation and the protein level of CHSY-1 were identified and quantified by Western blotting. Endothelin-1 caused time-dependent phosphorylation of Smad2C which was inhibited in the presence of the endothelin B receptor antagonist, BQ788. The response to ET-1 was inhibited by the Rho/ROCK kinase antagonist, Y27632 and by cytochalasin D, an inhibitor of actin polymerization but the ET-1-mediated pSmad2C was not inhibited by the matrix metalloproteinase (MMP) inhibitor, GM6001. ET-1 increased CHSY-1 protein level, which was inhibited in the presence of BQ788, cytochalasin D and Y27632. Endothelin-1 signalling via the ETB receptor utilizes cytoskeletal rearrangement and Rho kinase but not MMPs leading to TβRI transactivation signalling and phosphorylation of Smad2C and through this pathway increased the level of CHSY-1.


Palmitate induces DNA damage and senescence in human adipocytes in vitro that can be alleviated by oleic acid but not inorganic nitrate.
Hypertrophy in white adipose tissue (WAT) can result in sustained systemic inflammation, hyperlipidaemia, insulin resistance, and onset of senescence in adipocytes. Inflammation and hypertrophy can be induced in vitro using palmitic acid (PA). WAT adipocytes have innately low β-oxidation capacity, while inorganic nitrate can promote a beiging phenotype, with promotion of β-oxidation when cells are exposed to nitrate during differentiation. We hypothesized that treatment of human adipocytes with PA in vitro can induce senescence, which might be attenuated by nitrate treatment through stimulation of β-oxidation to remove accumulated lipids. Differentiated subcutaneous and omental adipocytes were treated with PA and nitrate and senescence markers were analyzed. PA induced DNA damage and increased p16INK4a levels in both human subcutaneous and omental adipocytes in vitro. However, lipid accumulation and lipid droplet size increased after PA treatment only in subcutaneous adipocytes. Thus, hypertrophy and senescence seem not to be causally associated. Contrary to our expectations, subsequent treatment of PA-induced adipocytes with nitrate did not attenuate PA-induced lipid accumulation or senescence. Instead, we found a significantly beneficial effect of oleic acid (OA) on human subcutaneous adipocytes when applied together with PA, which reduced the DNA damage caused by PA treatment.


Diffusion MRI in three types of anoxic encephalopathy.
We report the clinical and diffusion-weighted MRI (DWI) features of three patients with different types of anoxic brain injury: attempted hanging (hypoxic hypoxia), carbon monoxide poisoning (histotoxic hypoxia), and hanging with cardiac arrest (hypoxic-ischemic encephalopathy, HIE). The first two patients, but not the third, recovered substantial neurological function. The distribution of DWI abnormalities was different and correlated well with the distinct neuropathological features of these entities. The prognosis after anoxic encephalopathy depends on the underlying mechanism and its severity, which may be reflected by DWI abnormalities.


Body fluid compartment changes following neonatal surgery.
The neonate is born with an excess of total body water (TBW) in the range of 75% to 85% of body weight (v 60% in the adult), which is due to a large extracellular fluid volume (ECF) of 40% to 50% of body weight (v 20% in the adult). In an attempt to define the changes that occur in TBW and ECF following neonatal surgery, the following prospective study was carried out. Twenty newborns with major congenital anomalies (gastroschisis, 10; esophageal atresia, 4; ileal atresia, 3; omphalocele, 2; and malrotation, 1) were studied weekly for 4 weeks following surgery while being maintained on a standard protocol of total parenteral nutrition (TPN). Total body water was measured using deuterium oxide, and extracellular fluid volume was assayed with sodium bromide; both were expressed as percent body weight (BW). Weight gain or maintenance was observed in all infants. TBW remained unchanged (from 85.4% to 83.0%), and ECF decreased from 51.2% to 36.7% during the observation period. The TBW in the ten gastroschisis infants decreased from 87.3% to 78.0% v the ten other newborns, in whom there was no change (84.0% to 85.0%). The ECF changes were more dramatic, decreasing from 51.6% to 32.3% in the gastroschisis infants, v 50.8% to 45.5% in the other neonates (P = .0156). There were no differences between the two groups in the intake and output of fluids. The following conclusions were drawn: (1) Following major surgery, newborns show a steady decrease in ECF and minimal change in TBW in spite of adequate weight gain while receiving TPN.(ABSTRACT TRUNCATED AT 250 WORDS)


Skipping breakfast during pregnancy and hypertensive disorders of pregnancy in Japanese women: the Tohoku medical megabank project birth and three-generation cohort study.
Hypertensive disorders of pregnancy (HDP) adversely affect the prognosis of mother and child, and the prognosis depends on the subtype of HDP. Skipping breakfast may be associated with increased blood pressure due to disruption of the circadian clock, but the association with the development of HDP has not been studied. The purpose of this study was to examine the association between skipping breakfast and the development of HDP and HDP subtypes in Japanese pregnant women. Of the pregnant women who participated in the Tohoku Medical Megabank Project Three-Generation Cohort Study, 18,839 who answered the required questions were included in the analysis. This study had a cross-sectional design. The breakfast intake frequency from pre-pregnancy to early pregnancy was classified into four groups: daily, 5-6 times per week, 3-4 times per week, and 0-2 times per week. HDP was classified into gestational hypertension (GH), chronic hypertension (CH), preeclampsia (PE), and severe preeclampsia (SuPE). Multiple logistic regression analysis and multinomial logistic analysis were used to calculate odds ratios (ORs) and 95% confidence intervals (CIs) for breakfast intake frequency and development of HDP or HDP subtypes. We performed a stratified analysis based on energy intake. Of the participants, 74.3% consumed breakfast daily, and 11.1% developed HDP. Women who consumed breakfast 0-2 times per week had a higher risk of HDP (OR: 1.33, 95% CI: 1.14-1.56), CH (OR: 1.63, 95% CI: 1.21-2.19), and PE (OR: 1.68, 95% CI: 1.27-2.21) than those who consumed breakfast daily. No association was found between skipping breakfast and the risk of developing GH (OR: 1.26, 95% CI: 0.99-1.61) and SuPE (OR: 0.91, 95% CI: 0.55-1.49). Stratified analysis showed that the risk of developing HDP due to skipping breakfast was highest in the group with the highest daily energy intake. Skipping breakfast during pre-to early pregnancy is associated with the development of HDP. Further longitudinal studies are required to clarify the causal association between skipping breakfast and HDP.


The nerves of the juxtaglomerular apparatus of man and other mammals contain the potent peptide NPY.
The juxtaglomerular apparatus, a neuroendocrine unit located in the vascular pole of the glomerulus and influencing blood pressure by the secretion of renin, is known to have a rich supply of monoaminergic nerve fibres. Neuropeptide Tyrosine (NPY), a newly discovered, potent, vasoconstrictor peptide of 36 amino acids, has been found by immunocytochemistry to be present in a dense plexus of fibres around the juxtaglomerular apparatus of man, monkey, mouse, hamster, rat and guinea pig. NPY-immunoreactivity was markedly depleted after chemical sympathectomy by 6-hydroxydopamine. The concentration of NPY within the whole mouse kidney was 29.6 +/- 6.8 pmol/g and fractionation of the extracts demonstrated that the NPY-like immunoreactivity co-eluted from the column in the same position as the porcine NPY standard. The role of this peptide in renal physiology and pathology now needs urgent investigation.


Study of the Effect of Different Breast Implant Surfaces on Capsule Formation and Host Inflammatory Response in an Animal Model.
Breast implants are biomaterials eliciting a physiological and mandatory foreign body response. The authors designed an animal study to investigate the impact of different implant surfaces on the formation of the periprosthetic capsule, the inflammatory response, and the cellular composition. The authors implanted 1 scaled-down version of breast implants by different manufactures on 70 female Sprague Dawley rats. Animals were divided into 5 groups of 14 animals. Group A received a smooth implant (Ra ≈ 0.5 µm) according to the ISO 14607-2018 classification, Group B a smooth implant (Ra ≈ 3.2 µm), Group C a smooth implant (Ra ≈ 5 µm), Group D a macrotextured implant (Ra ≈ 62 µm), and Group E a macrotextured implant (Ra ≈ 75 µm). At 60 days, all animals received a magnetic resonance imaging (MRI), and 35 animals were killed and their capsules sent for histology (capsule thickness, inflammatory infiltrate) and immunohistochemistry analysis (cellular characterization). The remaining animals repeated the MRI at 120 days and were killed following the same protocol. MRI showed a thinner capsule in the smooth implants (Groups A-C) at 60 days (P < .001) but not at 120 days (P = .039), confirmed with histology both at 60 days (P = .005) and 120 days (P < .001). Smooth implants (Groups A-C) presented a mild inflammatory response at 60 days that was maintained at 120 days and a high M2-Macrophage concentration (anti-inflammatory). Our study confirms that smooth implants form a thinner capsule, inferior inflammatory infiltrate, and a cellular composition that indicates a mild host inflammatory response. A new host inflammatory response classification is elaborated classifying breast implants into mild, moderate, and high.


The association between prenatal psychosocial factors and autism spectrum disorder in offspring at 3 years: a prospective cohort study.
Few studies of risk factors for autism spectrum disorder (ASD) have been prospective in design or investigated the role of psychosocial factors measured during pregnancy. We aimed to investigate associations between prenatal psychosocial factors and risk of ASD in offspring, as part of a multicenter prospective cohort study of more than 2000 mother-child pairs. Nulliparous women aged 18-35 years, living in Pennsylvania, USA, were interviewed during pregnancy and multiple times postpartum over the course of a 3-year period. There were 2388 mothers who completed the Screen for Social Interaction Toddler Version (SSI-T), a measure of risk of ASD, when their child was 3-years old. Multivariable logistic regression models were used to investigate the associations between prenatal psychosocial factors-including total scores on three scales (social-support, stress and depression), trouble paying for basic needs, mental illness diagnosis and use of antidepressants-and risk of ASD in offspring at the age of 3-years, controlling for relevant confounding variables. There were 102 children (4.3%) who were scored as at-risk of ASD at 3-years. Prenatal psychosocial factors that were significantly associated with risk of ASD in the adjusted models were lower social-support (p < 0.001); stress (p = 0.003): depression (< 0.001), trouble paying for basic needs (p = 0.012), mental illness diagnosis (p = 0.016), and use of antidepressants (p < 0.001). These findings suggest that maternal experience of adverse psychosocial factors during pregnancy may be important intrauterine exposures related to the pathogenesis of ASD.


Cross-talk between cAMP and formylmet-leu-phe in human neutrophils: phosphorylation of a 52,000 molecular weight protein.
The mechanism of inhibition of neutrophil phagocytic functions by cAMP-elevating agents has not yet been clarified. In the present work, the effects of adenylate cyclase agonists on protein phosphorylation in the formylmethionyl-leucyl-phenylalanine (fMLP)-stimulated human neutrophils were studied. Before stimulation, 32Pi-labelled cells were incubated with adenosine deaminase to remove the endogenously produced adenosine, an adenylate cyclase agonist itself. A protein of about 52,000 molecular weight was rapidly and transiently phosphorylated when neutrophils were stimulated with fMLP in the presence of isoproterenol, prostaglandin E1, histamine or 2-chloroadenosine. This phosphorylation was blocked by the antagonists of the receptors for the above-listed agents. No phosphorylation of the 52,000 molecular weight protein could be observed if either fMLP or the cAMP-elevating agent were applied alone. A calcium ionophore A23187 and dibutyryl-cAMP could replace fMLP and a cAMP-elevating agent, respectively. Phosphorylation of the 52,000 molecular weight protein was also demonstrated in cell lysates in the presence of cAMP, and in membrane preparations in the presence of the catalytic subunit of cAMP-dependent protein kinase. These data suggest that phosphorylation of the 52,000 molecular weight protein in intact cells is dependent on the cross-talk between the fMLP- and the cAMP-signalling pathways, and may thus be involved in the cAMP-regulatory mechanism.


Evolution of the circuitry for conscious color vision in primates.
There are many ganglion cell types and subtypes in our retina that carry color information. These have appeared at different times over the history of the evolution of the vertebrate visual system. They project to several different places in the brain and serve a variety of purposes allowing wavelength information to contribute to diverse visual functions. These include circadian photoentrainment, regulation of sleep and mood, guidance of orienting movements, detection and segmentation of objects. Predecessors to some of the circuits serving these purposes presumably arose before mammals evolved and different functions are represented by distinct ganglion cell types. However, while other animals use color information to elicit motor movements and regulate activity rhythms, as do humans, using phylogenetically ancient circuitry, the ability to appreciate color appearance may have been refined in ancestors to primates, mediated by a special set of ganglion cells that serve only that purpose. Understanding the circuitry for color vision has implications for the possibility of treating color blindness using gene therapy by recapitulating evolution. In addition, understanding how color is encoded, including how chromatic and achromatic percepts are separated is a step toward developing a complete picture of the diversity of ganglion cell types and their functions. Such knowledge could be useful in developing therapeutic strategies for blinding eye disorders that rely on stimulating elements in the retina, where more than 50 different neuron types are organized into circuits that transform signals from photoreceptors into specialized detectors many of which are not directly involved in conscious vision.


The pharmacokinetics and tolerability of the oral neuraminidase inhibitor oseltamivir (Ro 64-0796/GS4104) in healthy adult and elderly volunteers.
The tolerability and pharmacokinetics of Ro 64-0802, a potent, selective inhibitor of influenza neuraminidase, and its oral prodrug oseltamivir were investigated in three double-blind, placebo-controlled studies. Two studies involved healthy adult volunteers (18-55 years) (n = 48) who received single (20-1000 mg) or bid doses (50-500 mg) (n = 32) of oseltamivir or placebo for 7 days. Healthy elderly volunteers (> or = 65 years) (n = 24) received oseltamivir 100 to 200 mg bid or placebo for 7 days in a third study. Measurable plasma concentrations of the active metabolite appeared rapidly in plasma and were significantly higher and longer lasting than those of oseltamivir. Pharmacokinetics of both compounds were linear. Multiple-dose exposure was predictable from single-dose data, and steady-state plasma concentrations were achieved within 3 days of bid drug administration. Oseltamivir was well tolerated at single doses of up to 1000 mg and twice-daily doses of up to 500 mg. Adverse events were mild in intensity. Exposure to both prodrug and active metabolite was increased in elderly patients by approximately 25%. However, due to the wide safety margin of both compounds, no dose adjustment is necessary for elderly patients.


Secretion of adenylate kinase 1 is required for extracellular ATP synthesis in C2C12 myotubes.
Extracellular ATP (exATP) has been known to be a critical ligand regulating skeletal muscle differentiation and contractibility. ExATP synthesis was greatly increased with the high level of adenylate kinase 1 (AK1) and ATP synthase beta during C2C12 myogenesis. The exATP synthesis was abolished by the knock-down of AK1 but not by that of ATP synthase beta in C2C12 myotubes, suggesting that AK1 is required for exATP synthesis in myotubes. However, membrane-bound AK1beta was not involved in exATP synthesis because its expression level was decreased during myogenesis in spite of its localization in the lipid rafts that contain various kinds of receptors and mediate cell signal transduction, cell migration, and differentiation. Interestingly, cytoplasmic AK1 was secreted from C2C12 myotubes but not from C2C12 myoblasts. Taken together all these data, we can conclude that AK1 secretion is required for the exATP generation in myotubes.


Effects of altering carbohydrate metabolism on energy status and contractile function of vascular smooth muscle.
Substrate-dependent changes in vascular smooth muscle energy metabolism and contractile function were investigated in isolated porcine carotid arteries. In media containing glucose glycogen catabolism accounted for all the estimated high-energy phosphate turnover that occurred in conjunction with contraction induced by 80 mM KCl. However, in glucose-free media glycogen catabolism accounted for only a portion of the estimated ATP utilization in resting and contracting arteries, even though glycogen stores were not depleted. The glycogenolysis and lactate production that ordinarily accompanies contraction was completely inhibited by 5 mM 2-deoxyglucose (2-DG). However, there was no decrease in the high-energy phosphate levels when compared to control resting arteries similarly treated with 2-DG. The results suggest that an endogenous non-carbohydrate source may be an important substrate for energy metabolism. Treatment of arteries with 50 microM iodoacetate (IA) in media containing glucose resulted in a marked reduction of high energy phosphate levels and an accumulation of phosphorylated glycolytic intermediates, as demonstrated by 31P-NMR spectroscopy. In glucose-free media, 50 microM IA had only a slight effect on high-energy phosphate levels, while glycogenolysis proceeded unhindered. With 1 mM IA in glucose-free media, the oxidative metabolism of glycogen was inhibited as evidenced by the depletion of high-energy phosphates and the appearance of sugar phosphates in the 31P-NMR spectra. Thus, the titration of enzyme systems with IA reveals a structural partitioning of carbohydrate metabolism, as suggested by previous studies.


Barriers and Facilitators to Improving Virginia's HPV Vaccination Rate: A Stakeholder Analysis With Implications for Pediatric Nurses.
Despite the evidence, the availability since 2006, and strong recommendations from many professional organizations, the human papillomavirus (HPV) vaccine has had a low uptake rate nationally and these trends have been even lower in the state of Virginia. We explored key stakeholders' perspectives on factors influencing HPV vaccination in central and southern Virginia organized within the socio-ecological model (SEM) framework. We conducted semi-structured key informant interviews with 31 stakeholders involved in HPV vaccination or cancer prevention. Stakeholders identified barriers at all SEM levels: Knowledge gaps and sexuality concerns (parent-child dyad level), time constraint and inconsistent recommendation (interpersonal level), lack of leadership and informational support (organizational and community level), and an ineffective mandate (policy level). Facilitators identified were realistic/receptive attitude (parent-child dyad level), provider's strong recommendation and educational support (interpersonal level), team approach and useful data (organizational level), educational outreach and community resources (community level), and support from federal and professional organizations (policy level). The stakeholder analysis provided an environmental scan of the barriers and facilitators so that an effective HPV vaccination strategy can be planned and implemented in the Commonwealth of Virginia by public health nurses.


Assay of sodium-lithium countertransport rate by direct measurement of erythrocyte lithium concentrations.
A method is described for the in-vitro measurement of erythrocytic sodium-lithium countertransport rate (SLC) by direct assay of intra-erythrocytic lithium (Li) concentrations in Li loaded cells during controlled Li efflux. Bicarbonate stimulated erythrocytic Li loading to an approximate intracellular concentration of 3 mmol/l can be achieved within 4 min. Maximum SLC rates are achieved at intra-erythrocytic concentrations greater than 2.5 mmol/l. Lithium efflux is linear over 0-60 min, and independent of pH change. The within-batch coefficient of variation for positive displacement pipetting of Li loaded erythrocytes is 2.8%. Storage of erythrocytes in isosmotic MgCl2 for 24 h had no effect on the SLC. Intra-individual coefficient of variation for SLC was less than 6%. The reference range for Caucasian subjects (n = 43) was 0.14-0.49 mmol Li +/l erythrocytes/h.


Characterization of the response of human bone marrow endothelial cells to in vitro irradiation.
Endothelial cell dysfunction is a classic consequence of radiation damage. Bone marrow endothelial cells (BMEC) are a critical component of the stroma in the regulation of haemopoiesis. In animal models, radiation-induced injury of BMEC has been described and a role for BMEC in haemopoietic regeneration after irradiation has been suggested. However, functions of BMEC involved in the haemopoietic regeneration have not been assessed. Therefore we studied the functional response of human BMEC to irradiation using the transformed human BMEC line (TrHBMEC) irradiated with 2. 5 or 10Gy. Our results showed a time- and a dose-dependent increase in damage to irradiated TrHBMEC measured by a decreased number of adherent cells which correlated with increased apoptosis and augmented release of soluble ICAM-1 and von Willebrand factor. 2 Gy irradiated TrHBMEC expressed more ICAM-1 on their surface than non-irradiated cells, whereas no change in VCAM-1, E-selectin and PECAM-1 expression was observed. An increased production of G-CSF, GM-CSF, IL-8, IL-6, IL-1alpha, IL-11, MIP-1alpha and SCF and no production of LIF, TNF-alpha, TPO and IL-3 by 2 Gy irradiated TrHBMEC was observed. The haemopoietic supportive function of TrHBMEC was not altered after a 2 Gy exposure. These results suggest that although radiation induces endothelial cell damage, irradiated cells still support the proliferation and the differentiation of CD34+ haemopoietic cells.


Immunohistochemical localization of immunoglobulins in bovine granulomatous lesions.
The immunohistochemical distribution of IgG, IgA and IgM in granulomatous lesions caused by Actinomyces bovis, Actinobacillus lignieresi, Actinomyces (Corynebacterium) pyogenes, Pseudomonas aeruginosa, Staphylococcus aureus and Mycobacterium bovis was studied. Numerous IgG-containing cells (plasma cells) were distributed in the peripheral connective tissue layers, but not in the epithelioid cell layer. A few scattered IgA- and IgM-containing cells were found in all the lesions examined. Mycobacterial granulomas contained fewer IgG-cells than did non-mycobacterial granulomas. Eosinophilic club-shaped bodies were found in A. bovis, A. lignieresi, P. aeruginosa and S. aureus, but they were generally negative for IgG, IgA and IgM.


N-ras mRNA expression is unaffected in glutathione-depleted cells of hematopoietic origin.
Glutathione (GSH) depletion in mitogen-stimulated T lymphocytes has been shown to markedly inhibit their proliferative response. This block in proliferation is associated with a significant reduction in total RNA and DNA synthesis; however, the specific mechanism involved in this inhibition of proliferation is unknown. Miller et al. have reported that lowering intracellular GSH levels by greater than 30%, in murine and human tumor cell lines of non-hematopoietic origin, leads to down-regulation of HA-, Ki- and N-ras oncogene expression [Miller. A.C., Gafner, J., Clark, E.P. and Samid, D. (1993) Mol. Cell Biol., 13, 4416-4422]. The reduction in ras transcript levels correlated with the extent of GSH depletion and was independent of the specific mode of oncogene activation. Since the activity of p21(ras) is thought to be involved in pathways of T cell activation, we set out to determine whether down-regulation of ras expression in T cells could be the mechanism by which T cell proliferation was inhibited in GSH-depleted T lymphocytes. Despite reducing the GSH level of concanavalin A-activated human peripheral blood mononuclear cells by 66%, no effect on ras mRNA expression was observed. Similarly, no reduction of ras transcript levels were detected in a human T cell line (Jurkat) or in a human monocytic cell line (THP-1) depleted of glutathione. Our results demonstrate that the mechanism by which GSH depletion inhibits T cell proliferation does not appear to involve a decrease in ras mRNA expression. In addition, our results suggest that differences in the regulation of ras mRNA expression may exist between lymphoid/monocytic cells of non-hematopoietic origin.


Loss characteristics of Cd in soil aggregates under simulated rainfall conditions.
Soil particles exert a significant influence on the migration of heavy metals in soil and water environments. In this study, a simulated rainfall experiment was conducted to investigate the loss characteristics of cadmium (Cd) from a red soil surface. Two rainfall intensities (50 and 100 mm h-1) were considered and the distribution of Cd in different fractions (>1, 1-0.25, 0.25-0.05 and <0.05 mm) of aggregates lost from the soil was examined. The result showed that in the artificially polluted soil, the Cd content in different aggregates decreased with particle size, with the lowest Cd content in the <0.05 mm fraction aggregates. In the rainfall experiment, the runoff rate generally increased with the rainfall duration, while the sediment yield rate first increased and then remained stable. The loss rates of various fractions of aggregates in the sediment generally followed the order of (<0.05) mm > 1-0.25 mm > 0.25-0.05 mm > (>1) mm. The proportions of the <0.05 mm-fraction aggregates were the highest in the loss sediment throughout the entire rainfall process. The lost Cd was transported mainly in the sediment-bound form, while only a very small portion of Cd was lost in the water phase of runoff. High rainfall intensity led to greater Cd loss in runoff. In the lost sediment, the concentration of Cd was higher in the macroaggregates. The phenomenon of Cd enrichment in the fine particles was not observed. Although the concentration of Cd in the <0.05 mm fraction was the lowest, the contribution of this fraction to the total Cd content in the sediments was the highest.


Pharmacological targeting of GLI1 inhibits proliferation, tumor emboli formation and in vivo tumor growth of inflammatory breast cancer cells.
Activation of the Hedgehog (Hh) pathway effector GLI1 is linked to tumorigenesis and invasiveness in a number of cancers, with targeting of GLI1 by small molecule antagonists shown to be effective. We profiled a collection of GLI antagonists possessing distinct mechanisms of action for efficacy in phenotypic models of inflammatory and non-inflammatory breast cancer (IBC and non-IBC) that we showed expressed varying levels of Hh pathway mediators. Compounds GANT61, HPI-1, and JK184 decreased cell proliferation, inhibited GLI1 mRNA expression and decreased the number of colonies formed in TN-IBC (SUM149) and TNBC (MDA-MB-231 and SUM159) cell lines. In addition, GANT61 and JK184 significantly down-regulated GLI1 targets that regulate cell cycle (cyclin D and E) and apoptosis (Bcl2). GANT61 reduced SUM149 spheroid growth and emboli formation, and in orthotopic SUM149 tumor models significantly decreased tumor growth. We successfully utilized phenotypic profiling to identify a subset of GLI1 antagonists that were prioritized for testing in in vivo models. Our results indicated that GLI1 activation in TN-IBC as in TNBC, plays a vital role in promoting cell proliferation, motility, tumor growth, and formation of tumor emboli.


30-year-old woman • progressive dyspnea and peripheral edema • 35th week of gestation with a history of mild preeclampsia • Dx?
► progressive dyspnea and peripheral edema ► 35th week of gestation with a history of mild preeclampsia.


Corrective exercises administered online vs at the workplace for pain and function in the office workers with upper crossed syndrome: randomized controlled trial.
To evaluate the effects of online-supervised versus workplace corrective exercises on neck-shoulder pain (NSP), sick leave, posture, workability, and muscular activity among office workers with the upper crossed syndrome (UCS). We performed a parallel-group randomized control trial at Shahid Beheshti University, Tehran, Iran, assigning 36 office workers to online-supervised, workplace, and control groups (mean (SD) age 38.91 ± 3.87, 38.58 ± 7.34, 37.00 ± 8.12). Inclusion criteria were alignment alteration (forward head (≥ 45°), rounding shoulder (≥ 52°), rounding back (≥ 42°), and pain intensity ≥ 3 in neck and shoulder. The two intervention groups performed 8-week exercise program, while the control group continued usual activities. Primary (NSP and sick leave) and secondary outcomes [postural angles, workability, and muscular activity were measured by VAS, outcome evaluation questionnaire (OEQ), photogrammetry, workability index, and EMG, respectively, at the baseline and an 8-week follow-up]. ANCOVA results revealed improvements for the online-supervised group versus control for NSP (P = 0.007), postural angles (P = 0.000, P = 0.001, P = 0.005), workability (P = 0.048, P = 0.042), and upper trapezius activation (P = 0.024, P = 0.016), respectively. Using paired t tests, both intervention groups improved from baseline to follow-up for NSP (P = 0.000, P = 0.002), forward head posture (P = 0.000, P = 0.000), round shoulders (P = 0.001, P = 0.031), and round back (P = 0.034, P = 0.008), respectively. Related parameters of workability (P = 0.041, P = 0.038), upper trapezius (P = 0.005, P = 0.005, P = 0.022), and serratus anterior (P = 0.020, P = 0.015) changed only in the online-supervised group. Online-supervised corrective exercise seems to improve a range of parameters related to work performance. These findings are highly applicable in light of the ongoing COVID pandemic; many workers have to work from home.


Gastrointestinal Endoscopy and the Risk of Prosthetic Joint Infection: A Nationwide Database Analysis.
Current guidelines suggest antibiotics prophylaxis is not necessary for patients with orthopedic prosthetics undergoing gastrointestinal endoscopy. Clinical evidence to support this recommendation is lacking. To analyze the association between inpatient gastrointestinal endoscopy and prosthetic joint infection (PJI) in patients with a recent arthroplasty. We included patients admitted from July to October of each calendar year (index admissions) who had an arthroplasty in the same calendar year prior to the index admission. We followed the occurrence of PJI for 60 days after the index admission. Only admissions from July to October were chosen as index admissions, and the follow-up period was limited to 60 days because the database structure prohibits the analysis of events in different calendar years. We compared the rate of 60-day PJI between those who had gastrointestinal endoscopy on index admissions to those who had not. We excluded patients aged less than 18 years, who died on index admission, or had any infection in the same calendar year before or during the index admission. Of 1,831,218 patients with arthroplasty, 88,345 met the inclusion criteria, out of which 5,855 had gastrointestinal endoscopy. The rate of 60-day PJI in those who had endoscopy was 0.23%, and in those who had not was 0.52% (P < 0.001). EGD without excision (adjusted odds ratio [95% confidence interval]: 0.20 [0.03-1.42], P = 0.107), EGD with excision (0.58 [0.21-1.60], P = 0.295), colonoscopy without excision (0.43 [0.11-1.72], P = 0.233), colonoscopy with excision (0.31 [0.04-2.21], P = 0.241), and PEG/PEJ (0.38 [0.05-2.71], P = 0.337) were not associated with risk of 60-day PJI. We found no PJI cases in patients underwent esophageal dilation, ERCP, and EUS with FNA. Gastrointestinal endoscopy in hospitalized patients with a recent previous arthroplasty is not associated with an increased risk of 60-day prosthetic joint infection.


Nanobodies: From Serendipitous Discovery of Heavy Chain-Only Antibodies in Camelids to a Wide Range of Useful Applications.
The presence of unique heavy chain-only antibodies (HCAbs) in camelids was discovered at Vrije Universiteit Brussel (VUB, Brussels, Belgium) at a time when many researchers were exploring the cloning and expression of smaller antigen-binding fragments (Fv and Fab) from hybridoma-derived antibodies. The potential importance of this discovery was anticipated, and efforts were immediately undertaken to understand the emergence and ontogeny of these HCAbs as well as to investigate the applications of the single-domain antigen-binding variable domains of HCAbs (nanobodies). Nanobodies were demonstrated to possess multiple biochemical and biophysical advantages over other antigen-binding antibody fragments and alternative scaffolds. Today, nanobodies have a significant and growing impact on research, biotechnology, and medicine.


Elevated serum lipids in hypogonadal men with and without hyperprolactinemia.
To determine whether men with hypogonadism are at risk for hyperlipidemia. Case-control study. Neuroendocrine clinical center of a referral-based university medical center. Consecutive sample of 18 men with testosterone deficiency who had prolactin-secreting pituitary adenomas, 15 men with acquired secondary hypogonadism and normal prolactin levels, and 33 normal male controls. We found a significant elevation in fasting cholesterol (6.23 +/- 0.28 mmol/L [mean +/- SE] compared with 5.17 +/- 0.13 mmol/L [241 +/- 11 mg/dL compared with 200 +/- 5 mg/dL], P less than 0.01), low density lipoprotein (LDL) cholesterol (4.11 +/- 0.23 mmol/L compared with 3.34 +/- 0.13 mmol/L [159 +/- 9 mg/dL compared with 129 +/- 5 mg/dL], P less than 0.05), and triglycerides (1.85 +/- 0.26 mmol/L compared with 1.11 +/- 0.07 mmol/L [164 +/- 23 mg/dL compared with 98 +/- 6 mg/dL], P less than 0.001) in men with hyperprolactinemia compared with controls. In the normoprolactinemic hypogonadal men, cholesterol (6.28 +/- 0.34 mmol/L [243 +/- 13 mg/dL], P less than 0.01), LDL cholesterol (4.34 +/- 0.34 mmol/L [168 +/- 13 mg/dL], P less than 0.01), and triglycerides (1.61 +/- 0.18 mmol/L [143 +/- 16 mg/dL], P less than 0.05) were also significantly higher than in the controls, and were the same as in the hyperprolactinemic men. High density lipoprotein (HDL) cholesterol did not differ among the three groups. Hypogonadism in men, with or without hyperprolactinemia, may be associated with elevation of fasting serum cholesterol, LDL cholesterol, and triglycerides compared with normal men. These data suggest that serum lipid levels should be evaluated in hypogonadal men. The presence of lipid abnormalities may affect the decision to treat testosterone deficiency in these patients.


Clinical and microbiological profiles of infective endocarditis in a tertiary hospital in Aseer region, Saudi Arabia.
We aimed to evaluate demographic data, underlying cardiac abnormalities, clinical profile, microbiological features, treatments and complications of infective endocarditis (IE) in a tertiary hospital in Aseer region, Saudi Arabia. A retrospective study of all cases with the diagnosis of definite endocarditis according to modified Duke Criteria admitted to ACH between May 2002 and April 2007. Data were reviewed on demographic and clinical data, underlying cardiac disease, microbiological findings, treatments and complications of IE. The study included 44 patients (28 males and 16 females; mean age 31.1 ± 16 years; range 13-65 years). Infective endocarditis developed on a native valve in 31 (70.5%), a mechanical prosthetic valve in 10 (22.7%), mitral valve prolapse in 2 (4.5%) and ventricular septal defect in 1 (2.3%). Rheumatic heart disease in 31 cases (70.5%) was the most common preexisting valvular abnormality in native valve endocarditis. The mitral valve was the most commonly affected valve 28 (63.6%). Fever occurred in 40 (90.9%) of the cases. Electrocardiography was abnormal in 34 cases (77.3%). Trans-thoracic and/or trans-esophageal echocardiography showed a vegetation in 22 (50%). Staphylococci in 10 cases (22.7%) and Streptococci in 8 cases (18%) were the most common causative agents and cultures were negative in 20 cases (45.5%). Twenty-two patients (50%) underwent surgical treatment. Congestive heart failure occurred in 16 (36.4%) cases, atrial fibrillation in 6 (13.6%) cases, and cerebrovascular accidents in 4 (9%) cases. Our data reflects the clinical and microbiological profiles of IE in a tertiary hospital in Aseer region, Saudi Arabia.


Vitamin B2-mediated cellular photoinhibition of botulinum neurotoxin A.
Botulinum neurotoxin (BoNT) is the most toxic species known to humans and has been identified as a potential bioterrorist threat. Unfortunately, the only existing countermeasures for BoNT intoxication involve vaccinations that are only effective prior to entry of the toxin into neuronal cells. Herein, we disclose the ability of the micronutrient riboflavin (vitamin B(2)) to photooxidatively inactivate BoNT in cell-based assays without the need for toxin and riboflavin pre-exposure. In total, this study suggests that botulism neurotoxicity may be blunted with photodynamic therapy technology.


The impact of intrathecal baclofen on the ability to walk: A systematic review.
Intrathecal baclofen (ITB) is an effective treatment for lower limb spasticity. In ambulatory patients with spasticity, there is a justifiable concern that ITB treatment may compromise ambulatory function by reducing the heightened muscle tone, thereby unmasking underlying muscle weakness. ITB is hence offered with reservation in ambulant patients. In this article, we review the literature surrounding the effect of ITB therapy on ambulatory function in patients with concurrent spasticity and discuss the key findings. A literature search of ProQuest Medline and EBSCO CINAHL databases was performed. Inclusion criteria included (a) studies reporting the effect of ITB in adult ambulatory patients; (b) studies with an intervention of screening test trial via either bolus injections or continuous infusion tests; and (c) studies with an intervention of ITB pump implantation. Seventeen eligible studies were identified and two authors independently assessed the study quality using the risk of bias in nonrandomised studies of interventions tool (ROBINS-I). Seventeen studies were included, with a total of 534 participants. Most of the patients remain ambulatory after ITB treatment, accompanied by improvements in gait speed and reduction in spasticity. ITB therapy when administered in carefully selected ambulatory patients with spasticity is not associated with loss of ambulatory function.


[Impact of COVID-19 pandemic on breast and gynecologic cancers management. Experience of the Surgery Department in the Nice Anticancer Center].
During the COVID-19 pandemic, the containment measures and the recommendations of several societies in oncology may have impacted the request for initial care for cancers. In this monocentric retrospective study, the number and the characteristics of patients received for a first consultation for a breast or gynecologic tumor were compared between the containment period and a control period. The times from diagnosis to treatment and the type of initial care were compared too. During the outbreak, 91 patients were seen for a new request, versus 159 during the control period, a decrease of 43.5 %. Patients were older (62.9 versus 60.9 years old) but this difference was not significant. Tumor stage was not modified. Concerning senology, the time from the biopsy to the first consultation was 5.5 days longer during the outbreak (difference statistically insignificant). Among the 51 patients requiring a surgical treatment during the outbreak, 16 (31.48 %) were postponed after the end of the containment measures. After all, the average time from the consultation to the treatment was not modified. No modification of type of treatment was observed. At the height of the pandemic, benefits and risks of the cancer treatment had to be daily balanced against the risk of exposition to the COVID-19. The evaluation of practices for cancer care is essential to understand the real impact of COVID-19 outbreak on global cancer management, so as to get prepared to further crises.


Microhabitat selection as an antipredator strategy in the aquatic insect Pachydiplax longipennis Burmeister (Odonata: Libellulidae).
An investigation of the larval dragonfly fauna associated with the plant, Sagittaria platyphylla, was conducted in a small pond. Despite the presence of several larval anisopteran species in the pond, only Pachydiplax longipennis larvae were found on Sagittaria plants. A study of the microspatial distribution of P. longipennis larvae on S. platyphylla indicated that larvae use the various regions of a plant in a highly non-random fashion. Larvae show a strong preference for the leaf axil area. A generalized predator, the bluegill sunfish (Lepomis macrochirus), was allowed to selectively eat either of two larvae placed in various plant regions. This experiment indicated that larvae in a leaf axil area were significantly less susceptible to bluegill predation than larvae positioned in other plant regions. The microspatial distribution of starved larvae revealed that larvae with high hunger levels occupied the leaf axil area significantly less than well fed larvae, suggesting 1) larvae do not use these regions as feeding sites, and 2) high hunger levels may induce a behavioral shift in habitat use, with starved larvae forced into areas of high predation risk by the need to fulfill nutritional requirements.


Technological trends in nanosilica synthesis and utilization in advanced treatment of water and wastewater.
Water and wastewater treatment applications stand to benefit immensely from the design and development of new materials based on silica nanoparticles and their derivatives. Nanosilica possesses unique properties, including low toxicity, chemical inertness, and excellent biocompatibility, and can be developed from a variety of sustainable precursor materials. Herein, we provide an account of the recent advances in the synthesis and utilization of nanosilica for wastewater treatment. This review covers key physicochemical aspects of several nanosilica materials and a variety of nanotechnology-enabled wastewater treatment techniques such as adsorption, separation membranes, and antimicrobial applications. It also discusses the prospective design and tuning options for nanosilica production, such as size control, morphological tuning, and surface functionalization. Informative discussions on nanosilica production from agricultural wastes have been offered, with a focus on the synthesis methodologies and pretreatment requirements for biomass precursors. The characterization of the different physicochemical features of nanosilica materials using critical surface analysis methods is discussed. Bio-hybrid nanosilica materials have also been highlighted to emphasize the critical relevance of environmental sustainability in wastewater treatment. To guarantee the thoroughness of the review, insights into nanosilica regeneration and reuse are provided. Overall, it is envisaged that this work's insights and views will inspire unique and efficient nanosilica material design and development with robust properties for water and wastewater treatment applications.


In vivo method for quantitation for anti-platelet potency of drugs.
Aortic strips from atherosclerotic rabbits or Achilles tendons from healthy rabbits were superfused with blood (3 ml/min) from anaesthetized and heparinized cats, while blood was returned to the venous system of animals. The superfused tissues gained in weight because of deposition of platelet thrombi on their surface. This gain in weight was continuously monitored and quantified. Forty minutes after intravenous administration of indomethacin (14 mg/kg), aspirin (7 mg/kg) or nictindole (2 mg/kg) the formation of platelet depostis was reduced by half. Three hours after i.v. administration of each drug at a dose of 20 m;/kg the remaining anti-platelet activities were 92% for aspirin, 59% for indomethacin and 18% for nictindole as compared to their antithrombotic action, which was recorded 40 min after their administration. Thrombogenesis was also prevented by a direct infusion of nictindole (50 ng/ml) or indomethacin (2000 ng/ml) into a stream of superfusing blood. Thereby our method enables us to quantify in vivo anti-aggregating potency of drug, to estimate the duration of this action, and to compare their in vitro and in vivo aggregation-inhibitory activities.


New HLA-DPB1 alleles generated by interallelic gene conversion detected by analysis of sperm.
The rate at which allelic diversity at the HLA loci evolves has been the subject of considerable controversy. The patchwork pattern of sequence polymorphism within the second exon of the HLA class II loci, particularly in the DPB1 locus, may have been generated by segmental exchange (gene conversion). We have analysed the frequency of variant DPB1 sequences that have been created by interallelic gene conversion in the germline by screening pools of sperm using PCR amplification and oligonucleotide probe typing. Our results indicate that about 1/10,000 sperm represents a new DPB1 sequence generated by short tracts of segmental exchange (gene conversion) within the second exon, suggesting that interallelic gene conversion may have an important role in generating the extensive allelic diversity at the HLA loci.


Navigating the Heterogeneity of Follicular Lymphoma and its Many Variants: An Updated Approach to Diagnosis and Classification.
Follicular lymphoma (FL) is a lymphoid neoplasm composed of follicle center (germinal center) B cells, with varying proportions of centrocytes and centroblasts, that usually has a predominantly follicular architectural pattern. Over the past decade, our understanding of FL has evolved significantly, with new recognition of several recently defined FL variants characterized by distinct clinical presentations, behaviors, genetic alterations, and biology. This manuscript aims to review the heterogeneity of FL and its variants, to provide an updated guide on their diagnosis and classification, and to describe how approaches to the histologic subclassification of classic FL have evolved in current classification schemes.


Transcatheter aortic valve implantation with the self-expandable venus A-Valve and CoreValve devices: Preliminary Experiences in China.
Transcatheter aortic valve implantation (TAVI) has been demonstrated to be an effective alternative to surgical aortic valve replacement (SAVR) in patients with aortic stenosis who are deemed high risk or inoperable. Currently, TAVI procedures in China mostly make use of the domestic Venus A-Valve and the CoreValve; however, there is no data on their comparative performance. Consecutive patients undergoing TAVI with the aforementioned devices were included. The outcomes were reported according to the Valve Academic Research Consortium-2 (VARC) definitions. A total of 54 TAVI procedures were performed, 27 with the CoreValve and the other 27 with the Venus A-Valve. An additional valve was required in 4 (14.8%) and 3 (11.1%) patients, and the VARC-2 device success rates were 81.5 and 85.2%, respectively. The incidences of common complications were similar, except for the significantly less frequent pacemaker insertion in the Venus A-Valve group (7.4 vs. 37.0%, P = 0.03). Within 30 days, 2 (3.7%) patients died, 1 in each group, and both had a bicuspid aortic valve (BAV). No other serious complications, such as annular rupture, coronary obstruction and aortic dissection, occurred in the 32 BAV patients treated with the Venus A-Valve (14) or the CoreValve (18). After 2-years of follow-up, there was no significant difference between CoreValve group and Venus-A group (11.1 vs. 7.4%, P = 0.64). TAVI with the domestic Venus A-Valve is feasible, safe, and can produce favorable short-term outcomes comparable to those with the CoreValve in inoperable or high-risk patients with tricuspid and bicuspid aortic valve stenosis. © 2017 Wiley Periodicals, Inc.


Bacteriochlorophyll aggregates in positively charged micelles.
Micellar complexes were prepared from bacteriochlorophyll a and bacteriopheophytin a with the cationic detergents, cetyltrimethyl ammonium bromide and cetylpyridinium chloride. These complexes have spectroscopic properties (absorption, circular dichroism) which are very different from the ones formed with non-ionic detergents like Triton X-100, and also with anionic detergents. Bacteriochlorophyll a forms two complexes: One is blue-shifted and has excitonically coupled Qy transitions. The second one is extremely red-shifted. The unusual properties are suggested to result from interactions of the positively charged head-group of the detergent with the tetrapyrrole.


Belief changes associated with psychedelic use.
Psychedelic use is anecdotally associated with belief changes, although few studies have tested these claims. Characterize a broad range of psychedelic occasioned belief changes. A survey was conducted in 2374 respondents who endorsed having had a belief changing psychedelic experience. Participants rated their agreement with belief statements Before and After the psychedelic experience as well as at the time of survey administration. Factor analysis of 45 belief statements revealed five factors: "Dualism," "Paranormal/Spirituality," "Non-mammal consciousness," "Mammal consciousness," and "Superstition." Medium to large effect sizes from Before to After the experience were observed for increases in beliefs in "Dualism" (β = 0.72), "Paranormal/Spirituality" (β = 0.90), "Non-mammal consciousness" (β = 0.72), and "Mammal consciousness" (β = 0.74). In contrast, negligible changes were observed for "Superstition" (β = -0.18).). At the individual item level, increases in non-physicalist beliefs included belief in reincarnation, communication with the dead, existence of consciousness after death, telepathy, and consciousness of inanimate natural objects (e.g., rocks). The percentage of participants who identified as a "Believer (e.g., in Ultimate Reality, Higher Power, and/or God, etc.)" increased from 29% Before to 59% After." At both the factor and individual item level, higher ratings of mystical experience were associated with greater changes in beliefs. Belief changes assessed after the experience (an average 8.4 years) remained largely unchanged at the time of survey. A single psychedelic experience increased a range of non-physicalist beliefs as well as beliefs about consciousness, meaning, and purpose. Further, the magnitude of belief change is associated with qualitative features of the experience.


Differentiation of Human Induced Pluripotent Stem Cells into Keratinocytes.
Investigating basic biological mechanisms underlying human diseases relies on the availability of sufficient quantities of patient cells. As most primary somatic cells have a limited lifespan, obtaining sufficient material for biological studies has been a challenge. The development of induced pluripotent stem cell (iPSC) technology has been a game changer, especially in the field of rare genetic disorders. iPSC are essentially immortal, can be stored indefinitely, and can thus be used to generate defined somatic cells in unlimited quantities. Further, the availability of genome editing technologies, such as CRISPR/CAS, has provided us with the opportunity to create "designer" iPSC lines with defined genetic characteristics. A major advancement in biological research stems from the development of methods to direct iPSC differentiation into defined cell types. In this article, we provide the basic protocol for the generation of human iPSC-derived keratinocytes (iPSC-K). These cells have the characteristics of basal epidermal keratinocytes and represent a tool for the investigation of normal epidermal biology, as well as genetic and acquired skin disorders. © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Directed differentiation of human iPSC into keratinocytes Support Protocol 1: Coating cell culture dishes or plates with Vitronectin XF™ Support Protocol 2: Freezing iPSC Support Protocol 3: Preparing AggreWell™ 400 6-well plates for EB formation Support Protocol 4: Coating cell culture dishes or plates with Collagen IV Support Protocol 5: Immunofluorescence staining of cells.


Viral causes of sudden inner ear deafness.
Clinical, pathologic, and laboratory studies suggesting a viral etiology for sudden deafness have been presented. Although a cause and effect relationship has not been proven, the frequent close association between viral infection and sudden deafness constitutes a strong argument in favor of a viral etiology of sudden deafness in about one of three patients with sudden deafness.


Mohs Micrographic Surgery for the Treatment of External Ear Melanoma: an Outcome Study.
The external ear is composed of thin skin overlying cartilage making melanoma on the external ear difficult to resect while preserving the intricate anatomy. Although surgeons have achieved robust clinical outcomes for nonmelanoma and most recently melanoma skin cancers with Mohs micrographic surgery (MMS), there is still not enough evidence on the MMS application for external ear melanoma treatment. The authors examined survival outcomes in patients treated with MMS, narrow margin excision (NME), and wide margin excision (WME) for melanoma on the external ear. Data from the NCI SEER program was retrospectively analyzed. Patients who received surgical treatment on the external ear and had microscopically confirmed diagnosis of cutaneous melanoma were included in the study. The effect of different surgery types: MMS, NME, and WME, on melanoma survival was evaluated. A total of 8,212 melanoma cases of the external ear performed during the years 2000 to 2015 were considered for analysis. There were no significant differences in survival comparing NME and WME with MMS. Mohs micrographic surgery is at least equivalent to WME for the treatment of melanoma of the external ear.


Cognition and health literacy in patients with hypertension.
Approximately half of the US population has marginal or inadequate health literacy, a measure highly associated with health outcomes. This measure is often linked to age and education, but recent evidence from patients with chronic heart failure suggests that much of age-related variability in health literacy can be explained by cognitive abilities (e.g., working memory, processing speed). We examined the role of cognitive and sensory abilities as mediators of age and education in determining functional health literacy among patients with hypertension. Four hundred ninety two community-dwelling adults diagnosed with hypertension (aged 21 to 92 years) participated. They were primarily female (73%), African-American (68%), and reported taking on average 7.8 prescribed medications. Before participation in a medication adherence intervention study, participants completed a battery of health literacy-related tasks. They completed tests that measured health literacy [Short Test of Functional Health Literacy in Adults (STOFHLA)], cognitive abilities (working memory, processing speed), sensory abilities (visual acuity and hearing), and physical health. Regression analyses showed that health literacy was related to age, education, and race (accounting for 24.4% of variance in STOFHLA scores). Cognitive ability accounted for an additional 24% of variance and greatly reduced the influence of age, education, and race (by 75%, 40%, and 48%, respectively). When controlling for cognitive and sensory variables, the association of age and education with STOFHLA scores was dramatically reduced. Thus, future interventions aimed at improving self-care for patients with low health literacy should aim to reduce demands on patients' cognitive abilities.


The DNA guanyl radical: kinetics and mechanisms of generation and repair.
The one-electron oxidation of DNA bases and single-stranded DNA was studied by pulse radiolysis of aqueous solutions from pH 7-7.4 at 20 degrees C. Thallic ions, Tl(II), were found to rapidly oxidize the purine nucleotides, deoxyguanosine 5'-monophosphate, k[Tl(II) + dGMP2-] = 3.4.10(9) M-1.s-1, and deoxyadenosine 5'-monophosphate, k[Tl(II) + dAMP2-] = 1.3.10(8) M-1.s-1. The reactivities of Tl(II) ions with model pyrimidine DNA bases, 1-methylcytosine and 1-methylthymine, were too low to be measured by pulse radiolysis, k less than 10(7) M-1.s-1. The Tl(II)-mediated oxidation of ssDNA, k = 2.8.10(8) M-1.s-1, produces DNA-guanyl radical, DNA-G.(-H), exclusively. The DNA-guanyl radical is found to be a potent oxidant in neutral media, E7 = 1.04 +/- 0.05 V. It rapidly oxidizes the aromatic amino acids in glycyl-tryptophan and tyrosine methyl ester, k = 3.6.10(7) M-1.s-1 and k = 1.7.10(8) M-1.s-1, respectively. These electron transfer processes indicate that a positive 'hole' may be transferred from DNA to a DNA-associated protein. The positive 'hole' in DNA can also be repaired by antioxidants, which are electron donors. The chemical repair of the DNA-guanyl radical by negatively charged antioxidants is slower than that by positively charged and neutral antioxidants.


Mechanisms and technology of marine oligosaccharides to control postharvest disease of fruits.
Fruit and vegetable postharvest diseases wreak havoc on food supply and the market economy. Meanwhile, the frequent use of synthetic fungicides poses a threat to the environment and public health. Marine oligosaccharides can act as an elicitor of postharvest disease resistance in fruits and also show broad-spectrum antimicrobial activity. Therefore, this review focuses on the mechanism and technology of marine oligosaccharides for postharvest disease control and compares them to horticulture studies to highlight the limitations of existing research and the prospects for future study. It is critical to developing marine oligosaccharides-based products for postharvest disease control.


Affinity purification and some properties of nucleoside diphosphatase from rat liver cytosol.
Nucleosidediphosphatase (nucleosidediphosphate phosphohydrolase, EC 188.8.131.52) of rat liver cytosol was purified up to 336--fold by the procedure including affinity chromatographies of concanavalin A- and alanine-Sepharose. The final purified enzyme showed a single protein band upon polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Its native form was found to be a tetramer with molecular weight of 120 000 which consists of subunit with molecular weight of 30 000. The enzyme was found to be a glycoprotein on the basis of its chromatographic behaviour with concanavalin A-Sepharose and positive staining with periodate-Schiff reaction in polyacrylamide gels. Furthermore, the two molecular forms with isoelectric points of 4.7 and 5.0 were demonstrated by electrofocusing, though they did not show any significant difference with respect to their catalytic properties.


Pollution exacerbates interregional flows of virtual scarce water driven by energy demand in China.
Existing studies on the virtual scarce water flows within the water-energy context focus on water quantity while largely ignoring water quality. This study improves the quantification method of scarce water uses by considering both blue water (representing water quantity) and grey water (indicating water quality). Based on a scarce-water extended multi-regional input-output model, we investigate the virtual scarce water flows driven by energy demand across 31 Chinese regions in 2012 and 2017. The results show that considering water quality provides new insights into the patterns of interregional flows of virtual scarce water driven by energy demand. The virtual integrated scarce water (VISW) flows, which consider both water quantity and quality, are 5 times the volume of virtual quantity-based scarce water (VQSW) flows. Moreover, certain regions (e.g., Hebei) are recognized as net VISW exporters, but are net importers in terms of VQSW. There are significant differences in the critical interregional pairs identified based on net VISW flows (e.g., Shandong-Zhejiang, and Shandong-Guangdong) and net VQSW flows (e.g., Heilongjiang-Guangdong, and Liaoning-Shaanxi). To reduce water scarcity based on the combined effect of both quantity and quality, the critical VISW interregional pairs should enhance cooperation through compensation payments and interregional technology transfer. This study highlights the importance of water quality in the assessment of virtual scarce water uses. Employing virtual scarce water as a policy tool to mitigate water scarcity might fail without the consideration of water quality.


Risk analysis of the spread of the quarantine pest mite Schizotetranychus hindustanicus in Brazil.
Schizotetranychus hindustanicus Hirst (Acari: Tetranychidae) known as the Hindustan citrus mite, is a quarantine pest present in Roraima, Brazil. In 1924 this pest was described in India. It was reported in 2002 in Venezuela and in Roraima in 2008. In 2010, the Hindustan citrus mite was reported in Colombia. It is possible that it will be introduced in other areas of Brazil, resulting in a threat to Brazilian citrus industry. Our objective was to determine the most suitable regions of Brazil for S. hindustanicus using a maximum entropy (Maxent) algorithm, based on native and invasive updated occurrence records from published research, field surveys and online databases. To avoid overfitting and improving transferability, we chose parameter settings of Maxent to construct and validate models by searching for the best combination of feature classes and regularization multipliers. The model obtained showed excellent performance according to all evaluation metrics used. A high potential for the establishment of S. hindustanicus was identified in large areas of Roraima, the extreme west of Amazonas, the entire north of the State of Pará, also in northeast, south, east, and north of the State of Amapá, and in a small portion northwest of the State of Maranhão (all states belonging to the northern region of Brazil). Our results provide information for policy making and quarantine measures, especially where S. hindustanicus is still absent in Brazil.


The second Ca2+-binding domain of the Na+ Ca2+ exchanger is essential for regulation: crystal structures and mutational analysis.
The Na(+)-Ca(2+) exchanger plays a central role in cardiac contractility by maintaining Ca(2+) homeostasis. Two Ca(2+)-binding domains, CBD1 and CBD2, located in a large intracellular loop, regulate activity of the exchanger. Ca(2+) binding to these regulatory domains activates the transport of Ca(2+) across the plasma membrane. Previously, we solved the structure of CBD1, revealing four Ca(2+) ions arranged in a tight planar cluster. Here, we present structures of CBD2 in the Ca(2+)-bound (1.7-A resolution) and -free (1.4-A resolution) conformations. Like CBD1, CBD2 has a classical Ig fold but coordinates only two Ca(2+) ions in primary and secondary Ca(2+) sites. In the absence of Ca(2+), Lys(585) stabilizes the structure by coordinating two acidic residues (Asp(552) and Glu(648)), one from each of the Ca(2+)-binding sites, and prevents a substantial protein unfolding. We have mutated all of the acidic residues that coordinate the Ca(2+) ions and have examined the effects of these mutations on regulation of exchange activity. Three mutations (E516L, D578V, and E648L) at the primary Ca(2+) site completely remove Ca(2+) regulation, placing the exchanger into a constitutively active state. These are the first data defining the role of CBD2 as a regulatory domain in the Na(+)-Ca(2+) exchanger.


Imaging Pattern of Diffuse Intrapulmonary Metastases in Lung Cancer Was Associated with Poor Prognosis to Epidermal Growth Factor Receptor Inhibitors.
Epidermal growth factor receptor (EGFR) mutations are more frequently seen in miliary intrapulmonary metastases than EGFR wild-type non-small cell lung cancer (NSCLC). Also, small-scale retrospective studies showed that patients harboring EGFR mutation with miliary pulmonary metastases had a worse prognosis. This study aimed to explore the impact of imaging patterns on the outcomes of EGFR tyrosine kinase inhibitor (TKI) treatment. A cohort of treatment-naive NSCLC patients harboring EGFR mutation with intrapulmonary metastases who were prescribed with TKI were enrolled. The demographic feature, clinical outcome, and CT imaging of each patient were reviewed and analyzed. A cohort of 174 patients were enrolled. Five intrapulmonary patterns of imaging were recognized: solid nodular, ground-glass nodular, miliary, multiple uniform nodular, and not otherwise specified. Among them, miliary and multiple uniform nodular patterns had similar poor prognosis, and, therefore, were combined as diffuse group. A worse PFS (9.0 mon, 95% CI: 8.0-10.0 mon) was observed compared with the rest (non-diffuse group, 13.3 mon, 95% CI: 10.2-16.4 mon, p<0.001, HR=0.49). The objective response rates (ORR) between the two groups were 76.8% and 84.1%, respectively, with no significant difference (p = 0.474). The OS of the diffuse and the non-diffuse group were 25.6 mon (95% CI 21.9-29.3 mon) and 35.0 mon (95% CI: 27.5-42.5, p = 0.01, HR= 0.59). Organs like bone (p=0.167), liver (p=0.513), and adrenal gland (p=0.375) were involved in similar frequencies in both groups. However, brain (p=0.070) and leptomeningeal (p=0.078) metastases were less common in the non-diffuse group with marginally statistical significance. The 2 groups contained similar missense mutations, and gene amplification was more common in the non-diffuse group. Patients with diffuse intrapulmonary metastases had inferior outcomes after TKI treatment. More aggressive treatments might be warranted for these patients.


Ketoconazole in the treatment of experimental murine paracoccidioidomycosis.
In a murine model of chronic disseminated paracoccidioidomycosis (strain 18; intravenous route), Ketoconazole (200 mg/kg in 0.2% agar) was given daily by gavage in three different schedules. Continuous treatment from an early stage of infection (day 3) up to week 20 was the most effective protocol, leading to remission of histopathological lesions and of both humoral and cellular anti-P. brasiliensis immune response, and clearance of the fungus in lungs; only 1 treated animal at week 20 showed pulmonary granulomas, although less extensive than control mice. Continuous treatment from early stage up to week 8, followed by a 16 week-period of drug discontinuity, caused remission of lesions in all but 3 treated mice which showed active pulmonary paracoccidioidomycosis similar to controls (14.2% of unresponsiveness to treatment). The continuous Ketoconazole protocol since a late stage of infection (week 4) up to week 20 produced a slower remission of lesions and immune response when compared with the first drug schedule. In this model of paracoccidioidomycosis, Ketoconazole showed no detectable side-effects and was a very effective drug especially in a prolonged administration protocol from an early stage of infection.


Characterization of inhibitory effects of perfluorooctane sulfonate on human hepatic cytochrome P450 isoenzymes: focusing on CYP2A6.
Perfluorooctane sulfonate (PFOS) is a chemically stable compound extensively used as oil and water repellent, surface active agents in our daily life. Accumulative research evidence gradually appears the toxicity of PFOS against mammals, but the whole figure remains to be elucidated. The present study was conducted to know the effects of PFOS on human hepatic drug metabolizing-type cytochrome P450 (CYP) isoenzymes such as CYP1A2 (7-ethoxyresorufin as a substrate), CYP2A6 (coumarin), CYP2B6 (7-ethoxy-4-trifluoromethylcoumarin), CYP2C8 (paclitaxel), CYP2C9 (diclofenac), CYP2C19 (S-mephenytoin), CYP2D6 (bufuralol), CYP2E1 (chlorzoxazone) and CYP3A4 (testosterone) in human livers employing their typical substrates. Although all of the oxidation reactions tested were more or less inhibited by PFOS, diclofenac 4'-hydroxylation mediated mainly by CYP2C9 was most strongly inhibited (K(i) value of 40 nM), followed by paclitaxel 6α-hydroxylation mediated mainly by CYP2C8 (K(i) value of 4 μM). The substrate oxidation reactions catalyzed by CYP2A6, CYP2B6, CYP2C19 and CYP3A4 were moderately (K(i) values of 35 to 45 μM), and those by CYP1A2, CYP2D6 and CYP2E1 were weakly inhibited by PFOS (K(i) values of 190-300 μM). The inhibition by PFOS for coumarin 7-hydroxylation mainly catalyzed by human liver microsomal CYP2A6 as well as by the recombinant enzyme was found to be enhanced by the preincubation of PFOS with human liver microsomes and NADPH as compared to the case without preincubation. The inhibition of the human liver microsomal cumarin 7-hydroxylation was PFOS concentration-dependent, and exhibited pseudo-first-order kinetics with respect to preincubation time, yielding K(inact) and K(I) values of 0.06 min(-1) and 23 μM, respectively. These results suggest that the metabolism of medicines which are substrates for CYP2C9 may be altered by PFOS in human bodies, and that PFOS is a mechanism-based inhibitor of CYP2A6.


Adolescent and parent perspectives on ethical issues in youth drug use and suicide survey research.
The contribution of adolescent and parent perspectives to ethical planning of survey research on youth drug use and suicide behaviors are highlighted through an empirical examination of 322 7th-12th graders' and 160 parents' opinions on questions related to 4 ethical dimensions of survey research practice: (a) evaluating research risks and benefits, (b) establishing guardian permission requirements, (c) developing confidentiality and disclosure policies, and (d) using cash incentives for recruitment. Generational and ethnic variation in response to questionnaire items developed from discussions within adolescent and parent focus groups are described. The article concludes with a discussion of the potential contributions and challenges of adolescent and parent perspectives for planning scientifically valid and ethically responsible youth risk survey research.


Detection rate of contrast-enhanced brain magnetic resonance imaging in patients with cognitive impairment.
The number of brain MRI with contrast media performed in patients with cognitive impairment has increased without universal agreement. We aimed to evaluate the detection rate of contrast-enhanced brain MRI in patients with cognitive impairment. This single-institution, retrospective study included 4,838 patients who attended outpatient clinics for cognitive impairment evaluation and underwent brain MRI with or without contrast enhancement from December 2015 to February 2020. Patients who tested positive for cognitive impairment were followed-up to confirm whether the result was true-positive and provide follow-up management. Detection rate was defined as the proportion of patients with true-positive results and was compared between groups with and without contrast enhancement. Individual matching in a 1:2 ratio according to age, sex, and year of test was performed. The overall detection rates of brain MRI with and without contrast media were 4.7% (57/1,203; 95% CI: 3.6%-6.1%) and 1.8% (65/3,635; 95% CI: 1.4%-2.3%), respectively (P<0.001); individual matching demonstrated similar results (4.7% and 1.9%). Among 1,203 patients with contrast media, 3.6% was only detectable with the aid of contrast media. The proportion of patients who underwent follow-up imaging or treatment for the detected lesions were significantly higher in the group with contrast media (2.0% and 0.6%, P < .001). Detection rate of brain MRI for lesions only detectable with contrast media in patients with cognitive impairment was not high enough and further study is needed to identify whom would truly benefit with contrast media.


Financial impact of energy efficiency and energy policies aimed at power sector reforms: mediating role of financing in the power sector.
This research examines how financial transformative power sector reforms affect energy efficiency and the economy in a sample of economies from South Asia, the Middle East, and Europe. We applied two stages of OLS, Bayesian VAR, and Data Envelopment Analysis (DEA) methods to a panel data set from 1995 to 2018. According to empirical findings, institutional deficiency has a negative effect on electricity reforms, implying that the greater the impact of reforms on electricity performance, the higher the institutional efficiency, A collection of reform initiatives involving a variety of reform agencies will boost energy efficiency by up to 13% and per capita electricity access by 62%. Despite recent reforms and regulatory measures, the electricity sector continues to face challenges in terms of private investment and structural flaws such as energy inefficiency, significant technological and financial losses, low power quality, and outdated transmission and network infrastructure. Interestingly 13.2% increases can be found in energy efficiency after electricity reforms. Unlike previous studies, our findings reveal a conflict between the broader economic effects and the welfare impact on electricity consumers.


[Effect of dibutyryl cyclic AMP on cerebral energy charge and electrocorticographic activity in rats under asphyxic hypoxia].
In Rats anesthetized with pentobarbital and submitted to brief asphyxic episodes, intracerebroventricular administration of dibutyryl cyclic AMP inhibits the decrease of energy charge, delays the disappearance of electrocorticographic activity (E Co G) and shortens the recovery time of E Co G.


Overactive bladder: prevalence and implications in Brazil.
To assess overactive bladder (OAB) prevalence, associated factors and implications in a young population. An independent population-based study was carried out. A self-applicative questionnaire covering urinary symptoms, coping strategies, quality of life and treatment seeking behavior was developed. A total of 848 subjects between 15 and 55 years completed the questionnaire. The overall prevalence of OAB was 18.9%. Women were significantly more affected than men (p = 0.001). All age groups were equally affected (p = 0.152). Subjects with OAB reported significant impairment on household chores (p = 0.009), physical activities (p = 0.016), sleep (p < 0.001), work (p < 0.001), social life (p < 0.001) and sexual life (p < 0.001). In addition, OAB individuals present higher prevalence of depression (p = 0.036), anxiety (p < 0.001), shame (p < 0.001) and tiredness (p < 0.001) OAB was independently associated to sexual life impairment (OR = 3.36, 95% CI=1.20-9.39). Only 27.5% of OAB subjects sought for medical counseling. OAB is a highly prevalent condition, even in such a young population. It affects both genders, yet it is more frequently observed in women. OAB is an important health condition, with serious impact on quality of life and sexual function. A large percentual of individuals remain unrecognized, under treated and consequently suffer for long periods of time.


Contribution of allelic variations to the phenotype of response to antidepressants and antipsychotics.
Individualized medicine through molecular pharmacogenetics is one of the major future goals in clinical medicine. In psychopharmacology, pharmacogenetics became an expanding research component. Major research results were already attained: first, it is now feasible to predict a major proportion of the interindividual variation of plasma levels of most antidepressants and antipsychotics by using the DNA-sequence variation in genes for crucial CYP P450-enzymes as CYP2D6. Second, it is now possible to relate serious side effects (tardive dyskinesia, weight gain) of antipsychotics to specific genetic variants of genes for target proteins. Third, a long list of mainly functional variants in target protein genes was explored for their predictive power for the beneficial and adverse treatment outcome. Although specific results transferable into clinical practice were not yet obtained in this respect, the proof of principle could be demonstrated.


A general expression for sequential DNA-fluorescence histograms.
A general expression for time sequences of DNA-fluorescence histograms from flow microfluorometry is given in this paper. Such expression is given in terms of the law of DNA growth along S phase, the flux of cells into S, and the efflux out of M. Special conditions of growth (e.g. the exponential steady-state growth), and the case of blocks in S phase are also analyzed. Some simulations of the expressions obtained are presented.


Potential risks of iatrogenic complications of nerve conduction studies (NCS) and electromyography (EMG).
Nerve conduction and electromyography studies are generally well tolerated and pose little risk to patients of serious adverse events in the hands of a well-trained competent practitioner. However, some patients and certain examinations do carry a higher risk of potential complications. It is good medical practice to inform patients of any risks, their potential severity and relative frequency. In order to obtain informed consent a dialogue should take place about the nature, purpose and effects of the studies, so patients can decide if they wish to undergo the proposed investigation. In this educational review we identify those procedures and patients at risk, and provide pragmatic practice recommendations for managing these material risks.


Alu elements mediate large SPG11 gene rearrangements: further spatacsin mutations.
Hereditary spastic paraplegias compose a group of neurodegenerative disorders with a large clinical and genetic heterogeneity. Among the autosomal recessive forms, spastic paraplegia type 11 is the most common. To better understand the spastic paraplegia type 11 mutation spectrum, we studied a group of 54 patients with hereditary spastic paraplegia. Mutation screening was performed by PCR amplification of SPG11 coding regions and intron boundaries, followed by sequencing. For the detection of large gene rearrangements, we performed multiplex ligation-dependent probe amplification. We report 13 families with spastic paraplegia type 11 carrying either novel or previously identified mutations. We describe a complex entire SPG11 rearrangement and show that large gene rearrangements are frequent among patients with spastic paraplegia type 11. Moreover, we mapped the deletion breakpoints of three different large SPG11 deletions and provide evidence for Alu microhomology-mediated exon deletion. Our analysis shows that the high number of repeated elements in SPG11 together with the presence of recombination hotspots and the high intrinsic instability of the 15q locus all contribute toward making this genomic region more prone to large gene rearrangements. These findings enlarge the amount of data relating repeated elements with neurodegenerative disorders and highlight their importance in human disease and genome evolution.


The lytic bacteriophage vB_EfaH_EF1TV, a new member of the Herelleviridae family, disrupts biofilm produced by Enterococcus faecalis clinical strains.
The aim of this study is to characterize a new bacteriophage able to infect Enterococcus faecalis, and to evaluate its ability to disrupt biofilm. The vB_EfaH_EF1TV (EF1TV) host-range was determined by spot test and efficiency of plating using a collection of 15E. faecalis clinical strains. The phage genome was sequenced with a next generation sequencing approach. Anti-biofilm activity was tested by crystal violet method and confocal laser scanning microscopy. Phage-resistant mutants were selected and sequenced to investigate receptors exploited by phage for infection. EF1TV is a newly discoveredE. faecalis phage which belongs to the Herelleviridae family. EF1TV, whose genome is 98% identical to φEF24C, is characterized by a linear dsDNA genome of 143,507 bp with direct terminal repeats of 1,911 bp. The phage is able to infect E. faecalis and shows also the ability to degrade biofilm produced by strains of this species. The results were confirmed by confocal laser scanning microscopy analyzing the biofilm reduction in the same optical field before and after phage infection. The EF1TV phage shows promising features such as an obligatory lytic nature, an anti-biofilm activity and the absence of integration-related proteins, antibiotic resistance determinants and virulence factors, and therefore could be a promising tool for therapeutic applications.


Usefulness of adenosine for arrhythmias in infants and children.
Adenosine was administered to 25 infants and children (11 patients after presenting with a sustained arrhythmia, and 14 during a diagnostic electrophysiologic study) to determine its electrophysiologic effects. Adenosine was given as an intravenous bolus (starting dose 37.5 micrograms/kg, and increased by 37.5 micrograms/kg increments until an effect was seen). Adenosine caused tachycardia termination or transient increased atrioventricular (AV) block in all 25 patients. Seven patients had tachycardia requiring only the atria for perpetuation and developed increased AV nodal block (minimum effective adenosine dose range 37.5 to 350 micrograms/kg, mean 131). Thirteen had AV reciprocating tachycardia or AV node reentry tachycardia (minimum effective adenosine dose range 37.5 to 225 micrograms/kg, mean 114). Four other patients received adenosine to rule out preexcitation (minimum effective adenosine dose range 37.5 to 375 micrograms/kg, mean 165). One of the 25 patients had junctional ectopic tachycardia and adenosine administration caused retrograde AV block. Six of the 25 (24%) had noticeable but minor side effects. One patient had sustained bradycardia (2 to 3 minutes requiring temporary pacing). Adenosine is a safe and effective agent in the evaluation and treatment of infants and children with arrhythmias.


BRCAness profile of sporadic ovarian cancer predicts disease recurrence.
The consequences of defective homologous recombination (HR) are not understood in sporadic ovarian cancer, nor have the potential role of HR proteins other than BRCA1 and BRCA2 been clearly defined. However, it is clear that defects in HR and other DNA repair pathways are important to the effectiveness of current therapies. We hypothesize that a subset of sporadic ovarian carcinomas may harbor anomalies in HR pathways, and that a BRCAness profile (defects in HR or other DNA repair pathways) could influence response rate and survival after treatment with platinum drugs. Clinical availability of a BRCAness profile in patients and/or tumors should improve treatment outcomes. To define the BRCAness profile of sporadic ovarian carcinoma and determine whether BRCA1, PARP, FANCD2, PTEN, H2AX, ATM, and P53 protein expression correlates with response to treatment, disease recurrence, and recurrence-free survival. Protein microarray analysis of ovarian cancer tissue was used to determine protein expression levels for defined DNA repair proteins. Correlation with clinical and pathologic parameters in 186 patients with advanced stage III-IV and grade 3 ovarian cancer was analyzed using Chi square, Kaplan-Meier method, Cox proportional hazard model, and cumulative incidence function. High PARP, FANCD2 and BRCA1 expressions were significantly correlated with each other; however, elevated p53 expression was associated only with high PARP and FANCD2. Of all patients, 9% recurred within the first year. Among early recurring patients, 41% had high levels of PARP, FANCD2 and P53, compared to 19.5% of patients without early recurrence (p = 0.04). Women with high levels of PARP, FANCD2 and/or P53 had first year cumulative cancer incidence of 17% compared with 7% for the other groups (P = 0.03). Patients with concomitantly high levels of PARP, FANCD2 and P53 protein expression are at increased risk of early ovarian cancer recurrence and platinum resistance.


Culture of human neoplastic gastrointestinal smooth muscle cells.
Due to limited growth potential of primary cultures and the absence of continuous lines of healthy enteric smooth muscle, we have studied the culture behavior of neoplastic gastrointestinal smooth muscle cells. Forty-six human enteric smooth muscle neoplasms (leiomyomas and leiomyosarcomas) were studied while fresh and/or after culture in vitro and growth in vivo in athymic nude mice, with assessments made of morphology, growth characteristics, and biochemical markers of differentiation. The state of differentiation of the tumors varied, with well-differentiated tumors tending to express binding sites for the gastrointestinal hormone cholecystokinin, whereas less well-differentiated tumors did not. Poorly differentiated tumors were the easiest to establish in culture in vitro and to grow in vivo in nude mice. When the cells placed directly into culture proliferated to confluent density, they underwent morphologic differentiation from a spread, fibroblastlike shape to a slender spindle morphology, with these cells possessing fewer biosynthetic organelles and arranging themselves in characteristic "hill and valley" arrays. However, the highly differentiated characteristics of expression of desmin or cholecystokinin-binding sites were not observed in cultured cells. In contrast, cells that had been passaged in nude mice before culture displayed a proliferative phenotype and failed to undergo morphologic differentiation on reaching confluent density. Four human enteric smooth muscle cell lines (documented by chromosomal analysis) originating in stomach, jejunum, ileum, and rectum were established using this strategy.


Results of new air caloric testing method among normal subjects. I. Biphasic testing.
A new air caloric testing method is described in which the temperature of a continuous aural irrigation is switched hot and cold values at times calculated to control the intensity of the resulting vestibular stimulation. Applications of low or high caloric stimulus intensities to normal subjects were well tolerated and reliably produced appropriate low or high intensity nystagmic responses. Nystagmus intensity values obtained from this study were compared with predicted intensity values from a computerized simulation of the actual test conditions, and also with values obtained when using biphasic water irrigations. As a result, further improvements in our methodology have been effected.


Clinical characteristics and long-term neurodevelopmental outcomes of leukomalacia in preterm infants and term infants: a cohort study.
Leukomalacia is a serious form of neonatal brain injury that often leads to neurodevelopmental impairment, and studies on neonatal leukomalacia and its long-term outcomes are lacking. The aim of this study was to analyze the clinical manifestations, imaging features, and long-term neurodevelopmental outcomes in preterm infants and term infants with leukomalacia. Newborns diagnosed with leukomalacia by head magnetic resonance imaging (MRI) and who were admitted to intensive care units from January 2015 to June 2020 were enrolled. All infants were followed up to June 2022 (2-7 years old), and their neurodevelopmental outcomes were evaluated. The clinical data and long- term outcomes of preterm infants and term infants was analyzed by Chi-square tests. A total of 218 surviving infants with leukomalacia including 114 preterm infants and 104 term infants completed the follow-up. The major typesof leukomalacia on MRI were periventricular leukomalacia in the preterm group and subcortical cystic leukomalacia in the term group, respectively (χ2 = 55.166; p < 0.001). When followed up to 2-7 years old, the incidence of neurodevelopmental impairment in the preterm group and term group was not significantly different (χ2 = 0.917; p = 0.338). However, the incidence of cerebral palsy (CP) in the preterm group was significantly higher (χ2 = 4.896; p = 0.027), while the incidence of intellectual disability (ID) (χ2 = 9.445; p = 0.002), epilepsy (EP) (χ2 = 23.049; p < 0.001), and CP combined with ID andEP (χ2 = 4.122; p = 0.042) was significantly lower than that in the term group. Periventricular leukomalacia mainly occurred in preterm infants while subcortical cystic leukomalacia was commonly seen in term infants. Although the long-term neurodevelopmental outcomes of leukomalacia were both poor, preterm infants were more prone to CP, while term infants were more prone to ID, EP, and the combination of CP with ID and EP.


Hepatic iron overload associated with a decreased serum ceruloplasmin level in a novel clinical type of aceruloplasminemia.
Aceruloplasminemia is a novel hereditary iron overload disease caused by a mutation in the ceruloplasmin gene and characterized by a complete deficiency of serum ceruloplasmin and iron accumulation in the liver and brain. We herein studied a novel clinical type of aceruloplasminemia in which a low amount of ceruloplasmin was detected in the serum of a patient. The patient presented with an asymptomatic hepatic iron overload, retinal degeneration, and diabetes mellitus. Magnetic resonance imaging of the liver and basal ganglia showed T2-hypointensity signals associated with parenchymal iron accumulation because of an absence of the ferroxidase activity in ceruloplasmin. A gene analysis showed a novel G969S mutation in the ceruloplasmin gene. A biochemical analysis of the patients' serum and a biogenesis study of G969S mutant ceruloplasmin using mammalian cell culture system resulted in the synthesis and secretion of only apoceruloplasmin without any ferroxidase activity. This novel clinical type of aceruloplasminemia should therefore be considered in the differential diagnosis of unexplained hemochromatosis, which is associated with a decrease in the serum ceruloplasmin level.


Abnormal cerebral blood flow and brain function in type 2 diabetes mellitus.
Type 2 diabetes mellitus (T2DM) lead to impaired cerebral blood perfusion, which leads to changes in brain function and affects the cognitive function of patients. In this study, cerebral blood flow (CBF) was used to evaluate the effect of T2DM on cerebral perfusion, and functional connectivity (FC) analysis was further used to explore whether the FC between the abnormal CBF region and the whole brain was changed. In addition, amplitude of low-frequency fluctuation (ALFF) and degree centrality (DC) were used to investigate the changes in spontaneous activity and connectivity strength of the brain network. We recruited 40 T2DM patients and 55 healthy controls (HCs). They underwent 3D-T1WI, rs-fMRI, arterial spin labeling (ASL) sequence scans and a series of cognitive tests. Cognitive test scores and brain imaging indicators were compared between the two groups, and the relationships among laboratory indicators, cognitive test scores, and brain imaging indicators were explored in the T2DM group. Compared to HCs, The CBF values of Calcarine_L and Precuneus_R in the T2DM group were lower. The DC value of Paracentral_Lobule_L and Precuneus_L, and the ALFF value of Hippocampus_L in the T2DM group were higher. In addition, the CBF values of Calcarine_L was negatively correlated with fasting insulin and HOMA_IR. This study found that there were regions of cerebral hypoperfusion in T2DM patients, which are associated with insulin resistance. In addition, we found abnormally elevated brain activity and enhanced functional connectivity in T2DM patients, which we speculated was the compensatory mechanism of brain neural activity.