Development of an animal model to study meshes used in genital prolapse surgery.
The objective was to develop an animal model using bacterial inoculation to evaluate tissue integration and tolerance to meshes used in genital prolapse surgery. We placed three different meshes under the abdominal skin of 120 Wistar rats: a polypropylene monofilament non-coated mesh (Parietene), a polypropylene monofilament collagen-coated mesh (Ugytex) and a polyethylene terephthalate mesh (Mersuture). We performed bacterial inoculation just after implantation with 1 ml of 10(7) colonies forming unit (CFU) of Staphylococcus epidermidis or Escherichia coli. Rats were sacrificed 7, 14, 60, and 90 days after intervention. We used polarised light microscopy to analyse the collagen deposition and organisation. We quantified the inflammation cells. Bacterial analysis and quantification of the explanted meshes were performed. The exact Fisher's test and Kruskal-Wallis test were used for statistics. We did not find any significant difference between inoculated or non-inoculated meshes in terms of collagen deposition. The scarring process seemed stable at day 90. Tissue integration was best with the polypropylene meshes, which allowed the development of a well-organised, mature connective tissue. Inflammatory reaction was higher in inoculated meshes, but only at day 7. At day 90, we found a high number of macrophages and multinuclear cells around all the meshes. There was no significant difference between prostheses that had been inoculated and those that had not with regard to positive bacterial culture. Quantification of bacterial colonies decreased with time. In this animal model, we did not find any clinically related difference in infection and tissue integration between the meshes used in genital prolapse. Such experimental studies must be carried out whenever new prostheses become available before their use is validated in common practice.


13C nuclear magnetic resonance and circular dichroism studies of the tuftsin conformation in water.
13C-NMR and circular dichroic (CD) spectra of tuftsin and its analogues are discussed in connection with our hypothesis that the beta-turn is the biologically active conformation of tuftsin. The changes in CD spectra evoked by an increase in pH are interpreted as a demonstration of the increasing amount of beta-turn conformers in solution. Configurational changes in successive residues of tuftsin showed that residues 2 and 3 of the peptide chain are important for the tuftsin conformation.


Persistent chromosome damage induced by localized radiotherapy for lymphoma.
A fibroblast culture was established from a lymph node biopsy of a patient with non-Hodgkin lymphoma, 9 months after chemotherapy and intensive therapeutic x-irradiation of the area. In contrast with blood and bone marrow, which were chromosomally normal, all cells of the lymph node were chromosomally abnormal, with numerous clones having multiple structural abnormalities. Numerical abnormalities (trisomies and monosomies) were not found. Structural abnormalities included translocations, terminal deletions, and pericentric inversions, with an excess of centromeric breakpoints being the only apparent deviation from a random distribution of breakpoints. None of the rearrangements associated with malignant lymphoma were seen, indicating that the chromosome abnormalities in the lymph stroma were radiation-associated, not disease-associated. These acquired changes may be a cause of additional malignant transformation.


IL-8 production in response to cigarette smoke is decreased in epithelial cells from COPD patients.
Cigarette smoke is the principal cause of chronic obstructive pulmonary disease (COPD), a disorder characterized by airway inflammation. As epithelial cells are the first line of defense against foreign material, the response of normal epithelial cells to smoke has been extensively studied. However, little is known about how epithelial cells derived from COPD patients respond to ongoing smoke exposure. This study was aimed at comparing the intracellular response of normal human bronchial/tracheal epithelial cells (NHBE) and COPD-diseased human bronchial/tracheal epithelial cells (DHBE) to cigarette smoke. NHBE and DHBE cells were treated with cigarette smoke condensate (CSC) for 24 h. IL-8 production was measured by ELISA and western blot was used to measure TLR4 expression. Cells were pretreated with CLI-095, a TLR4 inhibitor, or the signaling pathway inhibitors PD184352, Helenalin, or PI-103, which inhibit the ERK1/2, NF-κB and PI3K pathways, respectively. NHBE cells increased IL-8 production in a dose-dependent manner in response to CSC while DHBE cells did not show any significant difference and had a much lower production of IL-8 in response to CSC compared to NHBE cells. There was no change in TLR4 expression with CSC exposure. CLI-095 and PD184352 attenuated IL-8 secretion, indicating that CSC-induced inflammation is both TLR4- and ERK1/2-dependent. These results demonstrate that NHBE and DHBE cells differentially respond to cigarette smoke. DHBE cells exhibit a dampened IL-8 release, indicating that COPD is associated with a reduced capacity of airway epithelial cells to respond to foreign material.


Inhibition of conidiophore development in Aspergillus fumigatus by an Escherichia coli DH5α strain, a promising antifungal candidate against aspergillosis.
The opportunistic human pathogen Aspergillus fumigatus produces a massive number of asexual spores (conidia) as the primary means of dispersal, survival, genome protection and infection of hosts. In this report, we investigated secretory and cytosolic proteins of non-pathogenic bacterial species (mostly belonging to human microbiome) for antifungal potential against A. fumigatus, A. flavus and A. niger. Our preliminary results revealed that cytosolic proteins of E. coli DH5α were most active and the less toxic against various pathogenic isolates of A. fumigatus (the major pathogenic species), depicting a minimum inhibitory concentration (MIC) of 62.50 μg/mL, 62.50 μg/mL and 12.50 μg/disc using microbroth dilution assay (MDA), percentage spore germination inhibition assay (PSGI) and disc diffusion assay (DDA), respectively. E. coli protein was non-toxic against human erythrocytes at doses up to 1000 μg/mL as compared to standard drug, amphotericin B which lysed 100% of erythrocytes at a concentration of 37.50 μg/mL. Time kill analysis proved it to be fungicidal in a concentration and time-dependent manner. Scanning electron microscopic studies (SEM) were carried out to prevail what kind of damage it causes to A. fumigatus. SEM results reported that conidiophore (structures forming conidia) development was halted as a major consequence, reducing the number of conidiophores to insignificant values as well as alteration in their morphological attributes. This feature may contribute to the development of new prevention strategies against Aspergillus infections. Hyphal atrophy was also observed, evidenced by shrinking and flattening of hyphal walls and reduced, abrupt hyphal branching. Such actions may effectively reduce the invasive ability of Aspergillus as well as it can sterilize the fungal burden by obstructing the conidiation pathway of A. fumigatus. Hence, E. coli DH5α, being a commensal species, can lead to the development of antifungal molecule with novel targets in fungal metabolism, which will help in combating the antifungal resistance and toxicity associated with current therapy.


[COVID-19 unit hospitalization and family isolation: the value of a family information cell].
This worldwide health crisis forced hospitals to adapt their activities to this new confinement environment. Innovation was required to maintain a social bound between cohorting service's patients and their families, to guarantee a dedicated listening time and prevent as much as possible from the isolation due to this COVID-19 unusual situation. With that aim, the army training hospital Legouest created an information and support listening cell dedicated to COVID-19 hospitalized patients' families. It also provided several tablets for creating a patient and family digital gate.


Fluoxetine synergys with anticancer drugs to overcome multidrug resistance in breast cancer cells.
Multidrug resistance (MDR) is the main obstacle in breast cancer chemotherapy, a reversal reagent with high reversal effect but low toxicity is the hotpot issue at present. The antidepressant fluoxetine (FLX) is a new highly effective chemosensitizer; however, the possible mechanism of FLX in reversal of MDR is unclear. In this study, the effect of FLX on MDR mediated by apoptosis was researched in resistant/sensitive breast cancer cells, which treated by FLX/adriamycin (ADM)/paclitaxel (PTX) alone or FLX-ADM, FLX-PTX combination. Apoptosis assay demonstrated that FLX combined with ADM enhanced the proportion of apoptosis remarkably in MCF-7/ADM but not MCF-7 cells; however, increased the apoptosis rates in both cells when FLX-PTX combination. Results of apoptosis proteins assay showed a upgrade of p53 and a downgrade of Bcl-2 level by FLX-ADM or FLX-PTX combinations in both cells. Our findings indicated that by synergism with anticancer drugs, FLX modulation of apoptosis via targeting p53 and Bcl-2 expression, FLX reverse the breast cancer cell's resistance and enhance the chemosensitivity to ADM and PTX.


Recovery assessment of the branching coral Stylophora pistillata following copper contamination and depuration.
Most contemporary coral reefs live under both global (e.g. warming and acidification) and local (e.g. overfishing, pollution) stressors, which may synergistically undermine their resilience to thermal bleaching and diseases. While heavy metal toxicity in reefs has been well characterized, information on corals recovery from acute contamination is lacking. We studied for 42 days the ability of the coral Stylophora pistillata from the Gulf of Aqaba (northern Red Sea) to recover from a short (3 days) and prolonged (14 days) copper (Cu) contamination (1 μg L-1), after 11 ('Exp3/D11') and 28 ('Exp14/D28') days of depuration, respectively. Cu caused a decrease in chlorophyll content after 3 days, and in net photosynthesis (Pn) after 14 and 42 days. 'Exp14/D28' showed successful recovery based on Pn and relative electron transport rate, as opposed to 'Exp3/D11'. Results suggest the depuration time may be of greater importance than the exposure period to recover from such contamination.


Two antisense RNAs-AFAP1-AS1 and MLK7-AS1-promote colorectal cancer progression by sponging miR-149-5p and miR-485-5p.
Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths. Antisense RNAs (asRNAs) are closely associated with cancer malignancy. This study aimed to identify the action mechanism of asRNAs in controlling CRC malignancy. Analysis of the RNA sequencing data revealed that AFAP1-AS1 and MLK7-AS1 were upregulated in CRC patients and cell lines. High levels of both asRNAs were associated with poor prognosis in patients with CRC. Both in vitro and in vivo experiments revealed that the knockdown of the two asRNAs decreased the proliferative and metastatic abilities of CRC cells. Mechanistically, AFAP1-AS1 and MLK7-AS1 decreased the levels of miR-149-5p and miR-485-5p by functioning as ceRNAs. Overexpression of miRNAs by introducing miRNA mimics suppressed the expression of SHMT2 and IGFBP5 by directly binding to the 3' UTR of their mRNA. Knockdown of both asRNAs decreased the expression of SHMT2 and IGFBP5, which was reversed by inhibition of both miRNAs by miRNA inhibitors. In vivo pharmacological targeting of both asRNAs by small interfering RNA-loaded nanoparticles showed that knockdown of asRNAs significantly reduced tumor growth and metastasis. Our findings demonstrate that AFAP1-AS1 and MLK7-AS1 promote CRC progression by sponging the tumor-suppressing miRNAs miR-149-5p and miR-485-5p, thus upregulating SHMT2 and IGFBP5.


Therapeutical Potential of Imines; Synthesis, Single Crystal Structure, Computational, Molecular Modeling, and ADMET Evaluation.
Imines are multipurpose pharmacophores, simply accessible compounds, and have a broad range of usage in several areas of chemistry especially in medicine. Two novel compound imines, (E)-4-methyl-2-((o-tolylimino)methyl)phenol (1) and (E)-2-(((4-methoxybenzyl)imino)methyl)-4-methylphenol (2), were synthesized with effective product via reported protocol in the literature. Single crystal X-ray diffraction (SCXRD) was employed for structural exposition, disclosing that both compounds are orthorhombic. To optimize the newly designed imines, a B3LYP functional with a basis set 6-31G(d,p) was mainly considered. DFT results were utilized to check correlation between the data recovered from SCXRD outcomes and also to measure the energy difference. Hirshfeld surface study was done to demonstrate the intermolecular contacts along the percentage of interaction in the overall crystalline compound. Molecular operating environment program was tested against AChE and BChE enzymes to perform a modeling study of the compounds. The docking score and binding affinity of the compounds revealed that 2 showed comparatively more inhibition than 1. In silico ADMET studies exposed the physiochemical nature of these novel compounds, and it also unveiled that both compounds behaved as drug-like candidates.


Glucose 6-phosphate dehydrogenase is required for Salmonella typhimurium virulence and resistance to reactive oxygen and nitrogen intermediates.
Salmonella typhimurium zwf mutants lacking glucose 6-phosphate dehydrogenase (G6PD) activity have increased susceptibility to reactive oxygen and nitrogen intermediates as well as attenuated virulence in mice. Abrogation of the phagocyte respiratory burst oxidase during experimental infection with zwf mutant Salmonella causes a prompt restoration of virulence, while inhibition of inducible nitric oxide synthase results in delayed lethality. These observations suggest that G6PD-dependent bacterial antioxidant defenses play an important pathogenic role during early salmonellosis and additionally may help to antagonize NO-dependent antimicrobial mechanisms later in the course of infection.


High glucose-induced oxidative stress accelerates myogenesis by altering SUMO reactions.
Skeletal muscle preservation is a dynamic process that involves constant repair and regeneration. However, the regenerative capacity of muscle cells declines in hyperglycemia. This study aimed to explore the molecular mechanisms underlying this glucotoxicity during myoblast differentiation. C2C12 cells were exposed to different concentrations of glucose, to recapitulate the development of skeletal muscles in vivo in normo- and hyperglycemic conditions. In high glucose conditions, we found significant increases in levels of total cellular reactive oxygen species (ROS) and a reorganization of SUMO enzyme transcripts and SUMOylated proteins. Furthermore, in anticipation of the ROS-induced damage to newly formed myotubes, we observed acceleration of myogenesis. Interestingly, we found a tight relationship between SUMOylation of the Histone methyltransferase SET7/9 and the maintenance of sarcomeric structures of newly formed myotubes. Finally, treatment with the antioxidant anacardic acid preserved the function and activity of myotubes generated in high-glucose conditions by interfering with both ROS and SUMO pathways. Combined, these results suggest that increased oxidative stress and modulation of SUMO reactions are key mediators of glucotoxicity and inhibition of these perturbations using antioxidants might improve muscle regeneration in hyperglycemia.


Aneurysm Clip Compression Technique in the Surgery of Aneurysms with Hard/Calcified Neck.
One of the most important technical problems in surgery to repair aneurysms is the presence of a hard/calcified neck. In this situation, various techniques can be used for proper clipping. In addition to well-known techniques, we have used a different technique for more than 10 years. This technique probably also has been used by other neurosurgeons, but we could not find any publications about it in the English literature. Therefore, we would like to report the details of this technique and our own experience. More than 600 anterior circulation aneurysms were clipped between January 2003 and December 2014. It was necessary to apply this technique in 25 aneurysms because of a hard/calcified neck. This series was reviewed retrospectively. If the hard-calcified plaque at the neck does not allow for full closure of the clip and the known techniques are not sufficient for clipping, we carefully and slowly compress the aneurysm clip itself at the neck with a hemostatic clamp. The hard plaque usually is crushed with this technique, and full closure of the clip is immediately obtained. Complete occlusion of the neck was achieved in 16 aneurysms (64%) with this technique. Technique-related complication developed as intraoperative rupture of the aneurysm in two patients (8%). However, this complication was managed with other techniques in both cases. No distal thromboembolism developed in any patient. Our aneurysm clip compression technique may be a viable option in surgery of aneurysms with hard-calcified neck.


Reduction of plasma renin activity by inhibition of the fatty acid cyclooxygenase in human subjects: independence of sodium retention.
We carried out the present studies to determine whether the suppression of plasma renin activity (PRA) that follows inhibition of prostaglandin (PG) synthesis can be dissociated from the sodium-retaining effects of these drugs. In an initial investigation we studied the effect of indomethacin on PRA in normal subjects in balance on a 10 mM Na+ diet to prevent Na+ retention. Under these experimental conditions indomethacin did not lower PRA even though the fatty acid cyclooxygenase was inhibited, as indicated by a greater than 70% reduction in the major urinary metabolite of prostaglandin E (PGE-M). Sodium depletion leads to enhanced sympathetic activity. We therefore studied the effect of indomethacin on a group of subjects in 10 mM Na+ balance in whom the effect of increased beta-sympathetic activity was blocked by the administration of propranolol. In this group, indomethacin caused 65% suppression of PGE-M and had no effect on Na+ balance, but reversibly reduced PRA in the supine and upright positions by 84% and 70%, respectively. In normal subjects in 10 mM Na+ balance, the isoproterenol-induced increase in PRA also was unaffected by indomethacin. These data establish that inhibition of the cyclooxygenase can result in a reduction of PRA that is independent of changes in Na+ balance or beta-sympathetic tone.


Resistance to xenobiotics and parasites: can we count the cost?
The nature and cost of single genes of major effect is one of the longest running controversies in biology. Resistance, whether to xenobiotics or to parasites, is often paraded as an obvious example of a single gene effect that must carry an associated fitness 'cost'. However, a review of the xenobiotic resistance literature shows that empirical evidence for this hypothesis is, in fact, scarce. We postulate that such fitness costs can only be fully interpreted in the light of the molecular mutations that might underlie them. We also derive a theoretical framework both to encompass our current understanding of xenobiotic resistance and to begin to dissect the probable cost of parasite resistance.


Angiolymphoid hyperplasia with eosinophilia: report of a case and a review of the literature.
A case of angiolymphoid hyperplasia with eosinophilia mimicking multiple cylindromas (turban tumor) is presented in detailed clinical and histologic features, and the literature on the condition is reviewed.


Red blood cell transfusion dependence and outcome after allogeneic peripheral blood stem cell transplantation in patients with de novo myelodysplastic syndrome (MDS).
The prognosis of patients with de novo myelodysplastic syndrome (MDS) who are red blood cell transfusion-dependent (TD) and receive supportive care is inferior to that of patients who do not require transfusions. Whether TD also affects outcome after allogeneic transplantation is unknown. Consequently, in 172 de novo MDS patients (median age, 51 years), we analyzed the impact of TD on outcome after high-dose conditioning and allogeneic peripheral blood stem cell transplantation (PBSCT). With a median follow-up of 37 months, the probability of 3-year overall survival (OS) did not differ significantly between patients who were TD and those who were not TD before PBSCT (P=.1); however, transfusion burden, as reflected by ferritin levels, was correlated with a greater probability of severe acute graft-versus-host disease (aGVHD; P=.03) and a higher comorbidity index (P=.01), and OS was inferior in those patients with a ferritin level>1000 microg/L before PBSCT (P=.03). In multivariate analysis, only marrow myeloblast count (P=.01) and comorbidity index (P=.001) had a significant impact on OS. Our data do not identify TD as an independent negative prognostic factor for outcome after allogeneic PBSCT' however, iron overload (presumably transfusion-related) may contribute to poor transplantation success by adding to the overall comorbidities. Whether clinical intervention in the form of iron chelation can improve the outcome of allogeneic PBSCT in TD patients with MDS remains to be determined.


Transfection of cultured fish cells with exogenous DNA.
We have applied to fish cell cultures the techniques used routinely to introduce exogenous genes into cultured mammalian cells. Using calcium phosphate-mediated transfection, a plasmid containing bacterial aminoglycoside phosphotransferase under the influence of the simian virus 40 early promoter and polyadenylation signal was introduced into several fish cell lines. The plasmid was expressed in these cells in a stable manner, with transfection occurring at a frequency comparable to that seen with mammalian cells. These results suggest that plasmids constructed for use in mammalian cells may be used efficiently in fish systems without further modification and indicate that the advantages of transfection procedures utilized in mammalian systems can also be applied to fish.


Sensitivity and specificity of des-gamma-carboxy prothrombin for diagnosis of patients with hepatocellular carcinomas varies according to tumor size.
Serum levels of des-gamma-carboxy prothrombin (DCP) and alpha-fetoprotein (AFP) are known to be useful tumor markers for the diagnosis of hepatocellular carcinoma (HCC). The aim of this study was to examine the diagnostic efficacy of DCP and AFP in differentiating HCC from chronic liver diseases. We examined 1,377 HCC patients and 355 patients with chronic hepatitis or cirrhosis (non-HCC) who visited our institute and affiliated hospitals between June 1997 and September 2003. The median values of DCP and AFP were 60 mAU/mL and 34 ng/mL in HCC patients, respectively, and 18 mAU/mL and 3 ng/mL in non-HCC patients, respectively. The areas under the receiver operating characteristic (ROC) curves of DCP and AFP were 0.812 and 0.887, respectively (p < 0.0001). The area under the DCP ROC curve was significantly smaller than that of AFP in tumors less than 3 cm in diameter (p < 0.0001). However, the ROC area of DCP was significantly larger than that of AFP in tumors greater than 5 cm in diameter (p < 0.0001). The utility of DCP for the diagnosis of HCC was lower than that of AFP for small tumors, but higher than that of AFP for large tumors.


Structural basis for abrogated binding between staphylococcal enterotoxin A superantigen vaccine and MHC-IIalpha.
Staphylococcal enterotoxins (SEs) are superantigenic protein toxins responsible for a number of life-threatening diseases. The X-ray structure of a staphylococcal enterotoxin A (SEA) triple-mutant (L48R, D70R, and Y92A) vaccine reveals a cascade of structural rearrangements located in three loop regions essential for binding the alpha subunit of major histocompatibility complex class II (MHC-II) molecules. A comparison of hypothetical model complexes between SEA and the SEA triple mutant with MHC-II HLA-DR1 clearly shows disruption of key ionic and hydrophobic interactions necessary for forming the complex. Extensive dislocation of the disulfide loop in particular interferes with MHC-IIalpha binding. The triple-mutant structure provides new insights into the loss of superantigenicity and toxicity of an engineered superantigen and provides a basis for further design of enterotoxin vaccines.


Humanized substitutions of Vmat1 in mice alter amygdala-dependent behaviors associated with the evolution of anxiety.
The human vesicular monoamine transporter 1 (VMAT1) harbors unique substitutions (Asn136Thr/Ile) that affect monoamine uptake into synaptic vesicles. These substitutions are absent in all known mammals, suggesting their contributions to distinct aspects of human behavior modulated by monoaminergic transmissions, such as emotion and cognition. To directly test the impact of these human-specific mutations, we introduced the humanized residues into mouse Vmat1 via CRISPR/Cas9-mediated genome editing and examined changes at the behavioral, neurophysiological, and molecular levels. Behavioral tests revealed reduced anxiety-related traits of Vmat1 Ile mice, consistent with human studies, and electrophysiological recordings showed altered oscillatory activity in the amygdala under anxiogenic conditions. Transcriptome analyses further identified changes in gene expressions in the amygdala involved in neurodevelopment and emotional regulation, which may corroborate the observed phenotypes. This knock-in mouse model hence provides compelling evidence that the mutations affecting monoaminergic signaling and amygdala circuits have contributed to the evolution of human socio-emotional behaviors.


Synthesis of carotenoids by Rhodotorula rubra GED8 co-cultured with yogurt starter cultures in whey ultrafiltrate.
Two cultures, a yeast ( Rhodorula rubra GED8) and a yogurt starter ( Lactobacillus bulgaricus 2-11+ Streptococcus thermophilus 15HA), were selected for associated growth in whey ultrafiltrate (WU) and active synthesis of carotenoids. In associated cultivation with the yogurt culture L bulgaricus 2-11+S. thermophilus 15HA under intensive aeration (1.3 l(-1)min(-1) air-flow rate) in WU (45 g lactose l(-1)), initial pH 5.5, 30 degrees C, the lactose-negative strain R. rubra GED8 synthesized large amounts of carotenoids (13.09 mg l(-1 )culture fluid). The carotenoid yield was approximately two-fold higher in association with a mixed yogurt culture than in association with pure yogurt bacteria. The major carotenoid pigments comprising the total carotenoids were beta-carotene (50%), torulene (12.3%) and torularhodin (35.2%). Carotenoids with a high beta-carotene content were produced by the microbial association 36 h earlier than by Rhodotorula yeast species. No significant differences were notd in the ratio between the pigments synthesized by R. rubra GED8+ L. bulgaricus 2-11, R. rubra GED8+ S. thermophilus 15HA, and R.rubra GED8+yogurt culture, despite the fact that the total carotenoid concentrations were lower in the mixed cultures with pure yogurt bacteria.


Circulating endocannabinoid levels in pregnant women with gestational diabetes mellitus: a case-control study.
The role of the Endocannabinoids (ECs) in insulin resistance, and their association with visceral obesity and metabolic profile have been studied extensively. Since the association between ECs and metabolic factors in Gestational Diabetes Mellitus (GDM) are not clear, we aimed to evaluate the levels of N-Arachidonoylethanolamide (AEA) and 2-Arachidonoylglycerol (2-AG) and their association with C-reactive protein (CRP), glycemic indices, blood pressure, and anthropometric indices in pregnant women with GDM. The present case-control study was conducted among 96 singleton pregnant women aged 18-40 years, including 48 healthy pregnant women (control group) and 48 women with a positive diagnosis of GDM (case group). Odds Ratios (ORs) and 95% Confidence Intervals (CIs) for GDM were checked according to endocannabinoids and anthropometric indices using Multivariable Logistic Regression. AEA was significantly associated with increased risk of GDM in models 1, 2 and 3 (OR = 1.22, 95% CI: 1.06-1.41; OR = 1.54, 95% CI: 1.19-1.97; OR = 1.46, 95% CI:1.11-1.91). A positive but no significant association was found for AEA in model 4 (OR = 1.38,95% CI: 0.99-1.92). Similar to AEA, 2-AG was also positively associated with the likelihood of GDM in Models 1, 2, and 3 but the association attenuated to null in model 4 (OR = 1.25; 95% CI: 0.94- 1.65). Our findings showed that levels of ECs were significantly higher in pregnant women with GDM compared to healthy ones. Also, ECs levels were associated with the likelihood of GDM, independent of BMI and weight gain.


Cognitive mechanisms underlying depressive disorders in ADHD: A systematic review.
The risk for major depressive disorder (MDD) is considerably increased in young adults with attention-deficit/hyperactivity disorder (ADHD) but underlying mechanisms are poorly understood. This review explores ADHD-specific neurocognitive impairments as possible underlying mechanisms for ADHD-depression comorbidity. Two systematic literature searches were conducted in EBSCOhost, PubMED, and Cochrane Reviews databases according to PRISMA guidelines. The first search identified 18 meta-analyses of cross-sectional and longitudinal studies on cognitive dysfunctions in MDD across the lifespan. The second search identified six original studies on reaction time variability in MDD. During acute depression, children and adults showed cognitive deficits that overlapped with some of the ADHD-related impairments. Findings from remitted patients, high-risk individuals, and few prospective studies suggest that a subset of these shared impairments, specifically executive dysfunctions (selective attention, verbal fluency, working memory) and long-term memory problems, are candidate pre-existing risk markers of depression. We discuss if and how these specific neurocognitive mechanisms may mediate developmental pathways from ADHD to depression. If replicated by longitudinal studies, these findings may guide future prevention strategies.


Genetic determinants of diastolic and pulse pressure map to different loci in Lyon hypertensive rats.
Several genetic loci involved in blood pressure regulation have recently been localized in experimental models of hypertension, but the manner in which they influence blood pressure remains unknown. Here, we report a study of the Lyon hypertensive rat strain showing that different loci are involved in the regulation of steady-state (diastolic pressure) and pulsatile (systolic-diastolic, or pulse pressure) components of blood pressure. Significant linkage was established between diastolic blood pressure and a microsatellite marker of the renin gene (REN) on rat chromosome 13, and between pulse pressure and the carboxypeptidase B gene (CPB) on chromosome 2. These findings show that two independent loci influence different haemodynamic components of blood pressure, and that pulse pressure has a specific genetic determination.


Growth Rates of Marine Ciliates on Diverse Organisms Reveal Ecological Specializations within Morphospecies.
Abstract The growth rate of 31 stocks of Euplotes, a cosmopolitan, marine, unicellular protist, on six food species representing two different food types, microalgae and bacteria, has been determined. The 31 stocks represented nine reproductively isolated groups (biological species?) based on breeding relationships. Three morphospecies, E. vannus, E. crassus, and E. minuta, each with both autogamous and cross-breeding breeding groups, were included. The mean number of fissions completed in 5 days of a breeding group growing on one of the six food species varied from zero to 17.06. There is a strong interaction between morphospecies and food type. The largest morphospecies, E. vannus, translates the nutritional content of algae into growth better than it translates that of bacteria, while the reverse is true for E. minuta, the smallest morphospecies. Autogamous breeding groups grow more rapidly on algae than on bacteria when compared to cross-breeding groups in the same morphospecies. Two breeding groups cannot grow on Escherichia coli. ANOVA of fissions completed in 5 days revealed significant main effects and interactions between many hierarchical levels of stocks and food species. These significant interactions indicate that genetically determined ecologically important information is present at all taxonomic levels-morphospecies, breeding system, breeding group, and stock. As all these levels are biologically meaningful, measuring biodiversity in the E. vannus-crassus-minuta complex solely on morphospecies will inadequately represent the ecological diversity present in the organisms and their environment.


Left atrium reservoir function is central in patients with rheumatic mitral stenosis.
We investigated the relationship between Left Atrium strain reservoir function and symptoms and its impact on modulating Left Ventricular mechanics, diastolic filling, stroke volume, mean trans-mitral gradient and pulmonary pressure in mitral stenosis (MS) patients. We examined 195 full spectrum MS patients which were divided into two groups: Group 1 (n = 109) included patients with NYHA I & II functional class and group 2 (n = 86) included patients with NYHA III & IV functional class. LA strain reservoir function and classical echocardiographic parameters were calculated. LASr was significantly higher in group 1 versus group 2 in patients with MVA ≤ 1cm2 [8.8(6.0-12.6) vs 6.8(4.1-8.9), p = 0.03) and when 1cm2 < MVA ≤ 1.5 cm2 [10.0 (5.4-13.8) vs 6.7(4.5-9.0), p = 0.02). In patients with Pulmonary Hypertension, group 1 had significantly higher LASr than group 2 [11.1(6.6-14.8) vs 5.9(4.3-9.0), p = 0.002) By multivariate analysis, diabetes (OR = 4.11, 95%CI: 1.6-10.4), stroke (OR = 2.9, 95%CI: 1.1-7.9), LASr (OR = 0.9, 95%CI: 0.80-0.99) and LV ejection fraction (LVEF)(OR = 0.9, 95%CI: 0.91-0.99) were independently associated with NYHA functional class. LASr was significantly and positively correlated to MVA (r = 0.3, p < 10-3), stroke volume (r = 0.25, p = 10-3), mitral inflow (r = 0.4, p < 10-3) and LVEF(r = 0.14, p = 0.05). It was significantly and negatively correlated to left ventricular strain (r = -0.65, p < 10-3), LA indexed volume (r = -0.40, p < 10-3), maximum tricuspid regurgitation velocity (r = -0.25, p = 0.003), MTMG (r = -0.25, p = 10-3), and heart rate (r = -0.4, p < 10-3). We demonstrated a large range of interaction between LASr and mitral valve echocardiographic parameters. This may explain the reasons we identified LASr as an independent factor for MS functional tolerance.


Recurring priapism may be a symptom of voiding dysfunction - case report and literature review.
Recurring priapism is rare in pre-pubertal children and may be attributed to multiple causes. We propose that voiding dysfunction (VD) may also justify this symptom and detail a clinical case of recurring stuttering priapism associated to overactive bladder that completely resolved after usage of anticholinergics and urotherapy. Sacral parasympathetic activity is responsible for detrusor contraction and for spontaneous erections and a relationship between erections and bladder status has been proved in healthy subjects (morning erections) and models of medullar trauma. High bladder pressures and/or volumes, voiding incoordination and posterior urethritis can potentially trigger reflex erections.


Higher mycophenolate dosage is associated with an increased risk of squamous cell carcinoma in kidney transplant recipients.
Kidney transplant recipients are at increased risk of keratinocyte cancers, namely squamous cell and basal cell carcinomas (SCCs and BCCs). This is primarily due to the high levels of immunosuppression that are required to prevent allograft rejection. Different immunosuppressive medications confer different risks, and the effect of mycophenolate mofetil on SCC and BCC risk is unclear. We explored the relationship between mycophenolate dose prescribed over the entire transplant period and the risk of SCC and BCC. Kidney transplant recipients from Queensland, Australia, were recruited between 2012 and 2014 and followed until mid-2016. During this time transplant recipients underwent regular skin examinations to diagnose incident SCCs and BCCs. Immunosuppressive medication regimens were obtained from hospital records, and the average mycophenolate dose/day over the entire transplantation period was calculated for each patient. Doses were divided into three ranked groups, and adjusted relative risks (RRadj) of developing SCC and BCC tumours were calculated using negative binomial regression with the lowest dosage group as reference. Recipients who had used azathioprine previously were excluded; further sub-group analysis was performed for other immunosuppressant medications. There were 134 kidney transplant recipients included in the study. The average age was 55, 31% were female and 69% were male. At the highest median mycophenolate dose of 1818 mg/day the SCC risk doubled (RRadj 2.22, 95% CI 1.03-4.77) when compared to the reference group of 1038 mg/day. An increased risk persisted after accounting for ever-use of ciclosporin, ever-use of tacrolimus, and when excluding mammalian target of rapamycin users. This increased risk was mainly carried by kidney transplant recipients immunosuppressed for five or more years (RRadj = 11.05 95% CI 2.50-48.81). In contrast, there was no significant association between BCC incidence and therapy with the highest compared with the lowest mycophenolate dosage (RRadj = 1.27 95% CI 0.56-2.87). Higher mycophenolate dosage is associated with increased SCCs in kidney transplant recipients, particularly those immunosuppressed for more than five years. The increased SCC risk persists after accounting for usage of other immunosuppressant medications.


Cardiomyopathy in Soman and Sarin intoxicated rats.
Rats surviving various single dose of the organophosphorus anticholinesterase nerve agents Soman and Sarin were examined by light microscopy at intervals up to 35 days post-exposure. Brain lesions, identical to those that have been reported elsewhere were present, as well as a previously unreported finding associated with Soman or Sarin intoxication: half of all animals that had brain lesions also had areas of myocardial degeneration and necrosis. Depending upon the point in time at which cardiac tissues were examined, findings varied from areas of acute myolysis and necrosis to areas undergoing resolution of damage. The finding of brain lesions in those animals having cardiac lesions suggests a relationship between the convulsion induced neurologic and cardiac lesions. These studies suggest that convulsive doses of chemical warfare agents induce pathological changes in the cardiovascular system of laboratory animals.


Central nervous system manifestations of Mycoplasma pneumoniae infections.
Mycoplasma pneumoniae infection is associated with several manifestations from the central nervous system (CNS) such as encephalitis, aseptic meningitis, acute transverse myelitis, stroke, and polyradiculopathy. In the current paper epidemiologic, clinical, laboratory and treatment data on these manifestations are reviewed. The M. pneumoniae induced immune dysregulation and its contributing role in the pathogenesis of neurological insult is discussed. The recent introduction in clinical practice of newer molecular diagnostic techniques has helped in establishing a firmer association between M. pneumoniae infection and CNS disease especially encephalitis. Clinicians should be aware of the potential association between M. pneumoniae infection and several CNS manifestations. The role of various anti-microbial or immunomodulating therapies in treating such manifestations should be further explored.


Effects of diethylstilbestrol exposure in utero on the genital tracts of female ACI rats.
Aspects of development and morphology were studied in the reproductive tract of female ACI rats exposed prenatally to diethylstilbestrol (DES) and followed to 10 months of age. Pregnant ACI rats were injected with vehicle or DES (0.8 microgram = low DES or 8.0 micrograms = high DES) on Days 15 and 18 of gestation. At 12 weeks of age, half of the female offspring in each prenatal exposure group received a subcutaneous implant of a pellet containing 2.5 mg DES and 17.5 mg cholesterol; the remaining offspring received a control cholesterol pellet. Maternal reproductive performance was significantly impaired in DES-treated dams compared to controls. In female offspring mean time of vaginal opening was accelerated from 50.3 +/- 2.7 days in the vehicle-exposed group to 46.2 +/- 2.6 and 47.1 +/- 2.3 days in the low and high DES groups, respectively. Prior to pellet implantation, none of the rats exposed to DES prenatally was in "persistent estrus." At necropsy, rats exposed to DES in utero and implanted with the cholesterol pellet showed an increased frequency of atypical uterine epithelia, cystically dilated uterine glands, and a thickened vaginal epithelium. Among groups implanted with the DES pellet, prenatal exposure to DES increased the incidence of squamous metaplasia of the luminal epithelium and of cystically dilated uterine glands. Collectively, groups implanted with the DES pellet had higher incidences of squamous metaplasia of the uterine lumen, cystically dilated uterine glands, and patches of multilayered uterine epithelium than groups bearing the cholesterol pellet. DES pellet-bearing rats were also found to display a pronounced thickening and vacuolation of the vaginal epithelium. Cervical tissue from 98% of the DES-treated litters was characterized by a markedly convoluted epithelium with numerous squamous cell nests. There were no apparent effects of prenatal DES exposure or postnatal DES treatment on ovarian or oviductal histology. However, ovarian wet weights were significantly reduced as a result of postnatal DES treatment. Thus, the epithelial tissues of the uterus, cervix, and vagina in the ACI rat show a sensitivity to DES whether administered prenatally, postnatally, or in combination.


In vitro antileishmanial activity of trans-stilbene and terphenyl compounds.
Leishmaniasis are globally widespread parasitic diseases which often leads to death if left untreated. Currently available drugs present different drawbacks, so there is an urgent need to develop new, safe and cost-effective drugs against leishmaniasis. In this study we tested a small library of trans-stilbene and terphenyl derivatives against promastigote, amastigotes and intramacrophage amastigote forms of Leishmania infantum. Two compounds of the series, the trans-stilbene 3 and the terphenyl 11, presented the best activity and safety profiles. Terphenyl 11 showed a leshmanicidal activity higher than pentostam and the ability to induce apoptosis selectively in Leishmania infantum while saving macrophages and primary epithelial cells. Our data indicate that terphenyl compounds, as well as stilbenes, are endowed with leishmanicidal activity, showing potential for further studies in the context of leishmanial therapy.


Expression of PRL-3 regulates proliferation and invasion of breast cancer cells in vitro.
The protein tyrosine phosphatase PRL-3 plays an important role in cancer cell migration, invasion and metastasis. In breast cancer, PRL-3 is overexpressed in 70-75% of tumors and even more frequently in lymph node metastases. Moreover, PRL-3 overexpression in breast cancer is associated with an adverse disease outcome. Aim of this study was to determine the role of PRL-3 in breast cancer cell proliferation, migration and invasion in vitro. PRL-3 mRNA expression was evaluated in 6 breast cancer cell lines by quantitative real-time PCR. To investigate the effect of PRL-3 expression in breast cancer cells in vitro we both up- and downregulated PRL-3 expression in breast cancer cells and performed in vitro wound repair cell motility assays and invasion assays. The influence of PRL-3 knockdown in MCF-7 cells on the expression of several gene products involved in cell invasion and cytoskeletal function was evaluated with real-time PCR. PRL-3 mRNA expression was demonstrated in all breast cancer cell lines evaluated. Knockdown of PRL-3 in MCF-7 cells resulted in decreased proliferation, wound healing and invasion. PRL-3 knockdown in MCF-7 cells resulted in a significant reduction of heparanase, MMP-9, actin gamma-2 and Myosin 9 expression, and significant elevation of E-cadherin. We conclude that PRL-3 is an important regulatory factor for breast cancer cell proliferation and invasion. Loss of PRL-3 function induces an antimetastatic gene expression profile in breast cancer cells. Due to its role in tumor growth and metastasis, PRL-3 emerges as a new therapeutic target in breast cancer therapy.


Neutrophil to lymphocyte ratio and antidepressant treatment response in patients with major depressive disorder: Effect of sex and hippocampal volume.
Several factors may affect response to treatment in Major Depressive Disorder (MDD) including immune/inflammatory alterations and regional brain volumes, particularly in hippocampal regions which have shown to be influenced by inflammatory status. Neutrophil-to-lymphocyte ratio (NLR) is an inflammatory marker found to be elevated in depressed women in large population studies. Here we investigate the effect of NLR on treatment response in MDD patients, and the role of sex and hippocampal volume on influencing this relationship. A sample of 124 MDD depressed inpatients (F = 80) underwent MRI acquisition, admission NLR was calculated by dividing absolute neutrophil by absolute lymphocyte counts and depression severity was assessed at admission and discharge via the Hamilton Depression Rating Scale (HDRS). As a measure of treatment response, delta HDRS was calculated. We found a significant moderation effect of sex on the relationship between NLR and Delta HDRS: a negative relation was found in females and a positive one in males. NLR was found to negatively affect hippocampal volumes in females. Both left and right hippocampal volume positively associated with Delta HDRS. Finally, left hippocampal volume mediated the effect of NLR on Delta HDRS in females. Sex moderated the relation between inflammation and treatment response in line with previous reports linking inflammation to hampered antidepressant effect in females. Further, this effect is partially mediated by hippocampal volume, suggesting that antidepressant response may be hampered by the detrimental effect of inflammation on the brain.


Personal hygiene in external genitalia of healthy and hospitalized elderly women.
A comparative study of the personal hygiene of two groups of elderly women was carried out in Orebro, Sweden. The investigation group consisted of 28 women who were hospitalized in long-term care wards and the comparison group consisted of 35 healthy women. The results show that a remainder of soap in the genitalia may be one factor contributing to elderly hospitalized women's discomfort. Caregivers should be educated in how to assist elderly hospitalized women with their personal hygiene.


Risk factors and long-term outcomes of delayed graft function in deceased donor renal transplantation.
The purpose of this study was to analyze the risk factors for delayed graft function (DGF) and determine its impact on the outcomes of deceased donor (DD) kidney transplantation (KT). Between January 2000 and December 2011, we performed 195 DD renal transplants. After the exclusion of primary nonfunctional grafts (n = 4), the study recipients were divided into two groups-group I, DGF (n = 31, 16.2%); group II, non-DGF (n = 160, 83.8%). The following variables were compared: donor and recipient characteristics, patient and graft survival, postoperative renal function, acute rejection (AR) episodes, and the rates of surgical and infectious complications. Donor-related variables that showed significant differences included hypertension (P = 0.042), diabetes (P = 0.025), and prerecovery serum creatinine levels (P < 0.001). However, there were no significant differences in recipient-related factors. One significantly different transplant-related factor was positive panel reactive antibody (PRA > 20%, P = 0.008). On multivariate analysis, only the prerecovery serum creatinine level (P < 0.001; hazard ratio [HR], 1.814) was an independent risk factor for the development of DGF. A Cox multivariate analysis of risk factors for graft survival identified these independent risk factors for graft survival: nephron mass (donor kidney weight to recipient body weight ratio) index (P = 0.026; HR, 2.328), CMV infection (P = 0.038; HR, 0.114), and AR episode (P = 0.038; HR, 0.166). In DD KT, an independent risk factor for DGF was the prerecovery serum creatinine level. Although there was a significant difference in graft survival between the DGF and non-DGF groups, DGF was not an independent risk factor for graft failure in this study.


Re-face stereospecificity of methylenetetrahydromethanopterin and methylenetetrahydrofolate dehydrogenases is predetermined by intrinsic properties of the substrate.
Four different dehydrogenases are known that catalyse the reversible dehydrogenation of N5,N10-methylenetetrahydromethanopterin (methylene-H4MPT) or N5,N10-methylenetetrahydrofolate (methylene-H4F) to the respective N5,N10-methenyl compounds. Sequence comparison indicates that the four enzymes are phylogenetically unrelated. They all catalyse the Re-face-stereospecific removal of the pro-R hydrogen atom of the coenzyme's methylene group. The Re-face stereospecificity is in contrast to the finding that in solution the pro-S hydrogen atom of methylene-H4MPT and of methylene-H4F is more reactive to heterolytic cleavage. For a better understanding we determined the conformations of methylene-H4MPT in solution and when enzyme-bound by using NMR spectroscopy and semiempirical quantum mechanical calculations. For the conformation free in solution we find an envelope conformation for the imidazolidine ring, with the flap at N10. The methylene pro-S C-H bond is anticlinal and the methylene pro-R C-H bond is synclinal to the lone electron pair of N10. Semiempirical quantum mechanical calculations of heats of formation of methylene-H4MPT and methylene-H4F indicate that changing this conformation into an activated one in which the pro-S C-H bond is antiperiplanar, resulting in the preformation of the leaving hydride, would require a deltadeltaH(f) of +53 kJ mol-1 for methylene-H4MPT and of +51 kJ mol-1 for methylene-H4F. This is almost twice the energy required to force the imidazolidine ring in the enzyme-bound conformation of methylene-H4MPT (+29 kJ mol-1) or of methylene-H4F (+35 kJ mol-1) into an activated conformation in which the pro-R hydrogen atom is antiperiplanar to the lone electron pair of N10. The much lower energy for pro-R hydrogen activation thus probably predetermines the Re-face stereospecificity of the four dehydrogenases. Results are also presented explaining why the chemical reduction of methenyl-H4MPT+ and methenyl-H4F+ with NaBD4 proceeds Si-face-specific, in contrast to the enzyme-catalysed reaction.


Practice variation and missed opportunities for reperfusion in ST-segment-elevation myocardial infarction: findings from the Global Registry of Acute Coronary Events (GRACE).
The benefits of reperfusion therapy for patients with acute coronary syndromes have been established, but there is much variation in the type of reperfusion given and decisions about which patients are eligible. This study assessed current practices in relation to reperfusion therapy of ST-segment-elevation myocardial infarction from data collected in the multinational, prospective Global Registry of Acute Coronary Events. 94 hospitals in 14 countries are recruiting patients for the registry. Hospitals are organised into population-based clusters that reflect the population characteristics of the region. Information about patients' demographic characteristics, presenting symptoms, medical history, time between symptom onset and presentation, and clinical and electrocardiographic features is recorded in a standard case record. Of the 9251 patients enrolled, 1763 presented within 12 h of symptom onset with ST-segment-elevation myocardial infarction. Of these, 30% did not receive reperfusion therapy. Elderly patients (75 years and older), those presenting without chest pain, and those with a history of diabetes, congestive heart failure, myocardial infarction, or coronary bypass surgery were less likely to receive reperfusion therapy. The rate of primary percutaneous coronary intervention was highest in the USA and lowest in Australia, New Zealand, and Canada. The rate at sites with a catheterisation laboratory was 19% compared with zero at sites without this facility. A substantial proportion of patients who are eligible for reperfusion therapy still do not receive this treatment. These typically high-risk patients can be identified in advance, and reasons for the underuse of these beneficial treatments need to be clarified.


Comorbidity of impulse control disorders in pathological gamblers.
To investigate the rate of impulse control disorders among pathological gamblers and examine the relationship of comorbidity to gambling severity. Ninety-six adult pathological gamblers [mean age: 46.7 +/- 11.0 years; female: 44 (45.8%)] completed the following: Minnesota Impulsive Disorders Interview, Yale-Brown Obsessive-Compulsive Scale Modified for Pathological Gambling, and Gambling Symptom Assessment Scale. Twenty-two subjects (22.9%) reported a comorbid impulse control disorder, most commonly compulsive sexual behaviour and compulsive buying. Subjects with comorbidity reported significantly greater intensity of urges (t = -2.021; df = 94; P = 0.046) and thoughts (t = -2.147; df = 42.3; P = 0.038) related to gambling, and greater interference (t = -3.913; df = 48.1; P < 0.001) and distress (t = -2.504; df = 52.7; P = 0.015) secondary to gambling urges and thoughts. Impulse control disorders appear common among pathological gamblers and are associated with more severe gambling symptoms.


Predicting atrial fibrillation episodes with rapid ventricular rates associated with low levels of activity.
Rapid and irregular ventricular rates (RVR) are an important consequence of atrial fibrillation (AF). Raw accelerometry data in combination with electrocardiogram (ECG) data have the potential to distinguish inappropriate from appropriate tachycardia in AF. This can allow for the development of a just-in-time intervention for clinical treatments of AF events. The objective of this study is to develop a machine learning algorithm that can distinguish episodes of AF with RVR that are associated with low levels of activity. This study involves 45 patients with persistent or paroxysmal AF. The ECG and accelerometer data were recorded continuously for up to 3 weeks. The prediction of AF episodes with RVR and low activity was achieved using a deterministic probabilistic finite-state automata (DPFA)-based approach. Rapid and irregular ventricular rate (RVR) is defined as having heart rates (HR) greater than 110 beats per minute (BPM) and high activity is defined as greater than 0.75 quantile of the activity level. The AF events were annotated using the FDA-cleared BeatLogic algorithm. Various time intervals prior to the events were used to determine the longest prediction intervals for predicting AF with RVR episodes associated with low levels of activity. Among the 961 annotated AF events, 292 met the criterion for RVR episode. There were 176 and 116 episodes with low and high activity levels respectively. Out of the 961 AF episodes, 770 (80.1%) were used in the training data set and the remaining 191 intervals were held out for testing. The model was able to predict AF with RVR and low activity up to 4.5 min before the events. The mean prediction performance gradually decreased as the time to events increased. The overall Area under the ROC Curve (AUC) for the model lies within the range of 0.67-0.78. The DPFA algorithm can predict AF with RVR associated with low levels of activity up to 4.5 min before the onset of the event. This would enable the development of just-in-time interventions that could reduce the morbidity and mortality associated with AF and other similar arrhythmias.


What influences parents' decisions to limit or withdraw life support?
Decisions to forgo life support from critically ill children are commonly faced by parents and physicians. Previous research regarding parents' perspectives on the decision-making process has been limited by retrospective methods and the use of closed-ended questionnaires. We prospectively identified and described parents' self-reported influences on decisions to forgo life support from their children. Deeper understanding of parents' views will allow physicians to focus end-of-life discussions on factors important to parents and help resolve conflicts. Prospective, qualitative pilot study. Pediatric intensive care unit of a university-affiliated children's hospital. A total of 14 parents of ten children whose pediatric intensive care unit physician had made a recommendation to limit or withdraw life support. : In-depth, semistructured interviews were conducted with parents during their decision-making process. Factors influencing the parents in this study in their decision to forgo life support included their previous experience with death and end-of-life decision making for others, their personal observations of their child's suffering, their perceptions of their child's will to survive, their need to protect and advocate for their child, and the family's financial resources and concerns regarding life-long care. Parents in this study expressed the desire to do what is best for their child but struggled with feelings of selfishness, guilt, and the need to avoid agony and sorrow. Physician recommendations, review of options, and joint formulation of a plan helped parents gain a sense of control over their situation. Parents of eight children agreed to forgo life support and parents of two did not. Prospective interviews with open-ended questions identified factors influencing parents' decision making not previously described in the critical care literature such as parents' past experiences with end-of-life decisions and their anticipated emotional adjustments and future resources. Inclusion of these factors into discussions is important to parents and may facilitate decisions regarding the limitation or withdrawal of life support.


BRAF and RAS mutations in follicular variants of papillary thyroid carcinoma.
Follicular variants of papillary thyroid carcinoma (FVPTC), particularly the encapsulated subtype, often cause a diagnostic dilemma. Therefore, many FVPTCs are interpreted as "indeterminate" in preoperative fine-needle aspiration (FNA). The aim of this study was to analyze the genotypic changes in BRAF codons 600 and 601, as well as the N, H, and KRAS codons 12, 13, and 61 in FVPTCs and investigate the usefulness of preoperative BRAF and RAS mutation analysis as an adjunct diagnostic tool along with routine FNA. Surgically resected thyroid nodules were reviewed to establish the histological diagnosis of FVPTC. All preoperative FNA diagnoses were categorized according to the Bethesda Reporting System. Mutations in BRAF codons 600 and 601, and N, H, KRAS codons 12, 13, and 61 were analyzed by pyrosequencing. Of 132 cases, 81 (61.4 %) had a point mutation in one of the BRAF V600E, BRAF K601E, or RAS oncogenes; BRAF V600E in 43(32.6 %), BRAF K601E in three (2.3 %), and RAS in 35 (26.5 %) cases. All mutations were mutually exclusive. Of 78 cases with an FNA indeterminate category diagnosis, 51 (65.4 %) were positive for mutations: 24 for BRAF V600E, 3 for BRAF K601E, and 24 for the RAS gene. The KRAS mutation was more frequently found than the HRAS mutation, comprising 22.9 % of the RAS mutations, and all KRAS mutations were located at codon 61. This study demonstrated that either BRAF or RAS mutations were present in two thirds of FVPTCs and these mutations were mutually exclusive.


A pulmonary artery endothelial factor causes unidirectional alignment of smooth muscle cells.
Pulmonary artery and aorta endothelial and smooth muscle cells harvested from near term Rambouillet lambs were cultured. To determine whether cultured endothelial cells produce a factor which alters the shape and alignment of smooth muscle cells, rhodamine phalloidin staining of F-actin filaments was carried out, as well as time-lapse cinematography. We were able to demonstrate, with pulmonary artery but not with aortic cells in culture, an endothelial-derived factor which causes the smooth muscle cells to migrate and align in uniform direction. The effect could not be enhanced by making the endothelial cells hypoxic and was similar to that produced by exposing smooth muscle cells to KCl or directly to hypoxia.


Gastric emptying in labour: some recent findings and their clinical significance.
A résumé of the investigation of gastric emptying time in labour is presented with the results obtained recently in a series when the effects of narcotic analgesics, epidural analgesia, no analgesia and antagonistic drugs were studied. It is deduced that narcotic analgesics play a very large part in gastric delay in labour and epidural analgesia a comparatively small part. Some suggestions for future investigation and current management of the patient with delayed gastric emptying in labour are presented.


Assessment of the levels of polycyclic aromatic hydrocarbons and organochlorine contaminants in bottlenose dolphins (Tursiops truncatus) from the Eastern Atlantic Ocean.
The concentrations of 18 polychlorinated biphenyls (PCBs), 23 organochlorine pesticides (OCPs), and 16 polycyclic aromatic hydrocarbons (PAHs) were determined in the blubber and liver of 27 bottlenose dolphins (Tursiops truncatus) stranded along the Canary Islands coasts from 1997 to 2011. DDTs (mean of 60,960 and 445 ng/g lw., respectively) and PCBs (mean of 47,168 and 628 ng/g lw., respectively) were the predominant compounds in both tissues. Among PCBs the highly chlorinated PCB 180, 153 and 138 were the predominant congeners. We found a p,p'-DDE/∑DDTs ratio of 0.87 in blubber and 0.88 in liver, which is indicative of DDT ageing. All the samples showed detectable values of any of the 16 PAH studied. Phenanthrene was the most frequently detected and at the highest concentration. According to our results, concentrations of OCPs, and especially PCBs, are still at toxicologically relevant levels in blubber of bottlenose dolphins of this geographical area.


Pharmacokinetic, behavioral, and brain activity effects of Δ9-tetrahydrocannabinol in adolescent male and female rats.
Δ9-tetrahydrocannabinol (THC) is the intoxicating constituent of cannabis and is responsible for the drug's reinforcing effects. Retrospective human studies suggest that cannabis use during adolescence is linked to long-term negative psychological outcomes, but in such studies it is difficult to distinguish the effects of THC from those of coexisting factors. Therefore, translationally relevant animal models are required to properly investigate THC effects in adolescents. However, though the relevance of these studies depends upon human-relevant dosing, surprisingly little is known about THC pharmacology and its effects on behavior and brain activity in adolescent rodents-especially in females. Here, we conducted a systematic investigation of THC pharmacokinetics, metabolism and distribution in blood and brain, and of THC effects upon behavior and neural activity in adolescent Long Evans rats of both sexes. We administered THC during an early-middle adolescent window (postnatal days 27-45) in which the brain may be particularly sensitive to developmental perturbation by THC. We determined the pharmacokinetic profile of THC and its main first-pass metabolites (11-hydroxy-THC and 11-nor-9-carboxy-THC) in blood and brain following acute injection (0.5 or 5 mg/kg, intraperitoneal). We also evaluated THC effects on behavioral assays of anxiety, locomotion, and place conditioning, as well as c-Fos expression in 14 brain regions. Confirming previous work, we find marked sex differences in THC metabolism, including a female-specific elevation in the bioactive metabolite 11-hydroxy-THC. Furthermore, we find dose-dependent and sex-dependent effects on behavior, neural activity, and functional connectivity across multiple nodes of brain stress and reward networks. Our findings are relevant for interpreting results of rat adolescent THC exposure studies, and may lend new insights into how THC impacts the brain in a sex-dependent manner.


Insurance Type and Marital Status Impact Hospital Length of Stay After Pancreatoduodenectomy.
Recognition of the impact of social determinants on health care and surgical outcomes is imperative to improve patient care. This study aims to examine the impact social determinants have on hospital length of stay (LOS) after pancreatoduodenectomy (PD). Retrospective review of a prospective American College of Surgeons-National Surgical Quality Improvement Program database identified patients who underwent PD from 2013 to 2018. Patients were categorized by insurance type (public/private/multiple), and electronic medical record review was performed to obtain distance from home, marital status, and race. Public insurance included Medicare and Medicaid; multiple types were defined as public insurance supplemented by a private insurance. Univariable analysis was used to identify potential confounders. Significant differences (P < 0.05) were controlled for using multivariable regression models to examine the effect of variables on LOS. About 813 PDs were included (n = 341 public; n = 238 private; and n = 234 multiple). Patients with public insurance had significantly longer LOS than patients with private on univariate (P < 0.001) and multivariable analyses (P = 0.021) (8 versus 7 d). Patients with multiple insurance types showed significantly increased LOS compared with patients with private on univariable (P < 0.001) and multivariable analyses (P = 0.006) (8 versus 7 d). Single patients had significantly longer LOS compared with married patients on univariable (P = 0.012) and multivariable analyses (P = 0.005) (8 versus 7 d). Distance from home, race, gender, or age did not have a significant impact on LOS. Single patients and patients with public or multiple insurance types are more likely to have longer hospital LOS after PD. These findings will enable physicians to identify patients at risk and target them for enhanced recovery programming.


Towards the structure of the mammalian signal recognition particle.
The signal recognition particle (SRP) is a ubiquitous ribonucleoprotein particle involved in the co-translational targeting of proteins to membranes. Crystal structures are now available for three protein-RNA subcomplexes from the SRP, which give insights into fundamental aspects of protein-RNA recognition, the assembly of stable ribonucleoprotein particles and the mechanism of action of the SRP.


Hearing loss on social media: Who is winning hearts and minds?
To analyze specific patterns of Twitter usage using common references to hearing loss, and characterize the virtual public that comprises the hearing loss community to inform hearing loss stakeholders for opportunities for engagement and outreach. Social media network analysis. Twitter tweets were sampled from July 2016 to September 2016 using #hearing, #hearingloss, #deaf, #hearingimpairment, #hardofhearing, #deafness, #hearingmatters, #hearinghealth, and #hearingimpaired tags. User and Twitter social community metrics were examined including temporal trends, tweet content, user activity, tweet reach, and an analysis of the tweets' social network. We identified and analyzed 49,208 tweets from July 2016 to September 2016 with tags relevant to hearing loss. Of the 100 most active Twitter accounts, organizations owned 67% compared to 33% owned by individuals. Commercial/for profit and informational organizations were the most common organization account owners (26% and 16%, respectively). Five unique tweets were identified as each having a reach of over 100,000 Twitter users, with the greatest reach exceeding 250,000 users. Temporal analysis identified marked retweet outliers (>300 retweets per hour) that corresponded with a widely publicized event involving the dismissal of a deaf employee from a fast-food chain store. Twitter accounts owned by organizations outnumbered individual accounts, and commercial/for profit user accounts were the most frequently active organization account type. Tweets pertaining to hearing loss may have a broad reach to a large community base. Analyses of social media use can be helpful in discovering issues of interest to the hearing loss community, as well as determining which users and organizations are dominating social network conversations. NA. Laryngoscope, 128:1453-1461, 2018.


The radiation-releasable cell wall nuclease of Micrococcus radiodurans. Purification and properties of the native enzyme.
Micrococcus radiodurans is known to possess a surface nuclease located in a mid-wall layer. Previous work showed that hydroxyl radicals, generated by sublethal doses of ionizing radiation, attack the cell wall and initiate release of the active enzyme into the external medium. The enzyme from unirradiated cells has now been purified to homogeneity by a simple 3-step process. The nuclease is a dimer of molecular weight about 260 000 and exonucleolytically degrades both DNA and RNA to 5'-mononucleotides. Single-stranded DNA is degraded at a rate about 200 times faster than double-stranded DNA. Oligonucleotides bearing a terminal 3'-phosphate are resistant to digestion. Dinucleotides must possess a 5'-terminal phosphate and a free 3'-terminal OH to be hydrolyzed. The enzyme has a pH optimum of about 9.0. A metal ion, possibly Ca2+, appears to be tightly bound to the protein. Removal of this metal with EDTA inactivates the enzyme. Simple readdition of Ca2+ does not restore activity although partial function can be recovered by renaturation from urea in the presence of this ion. The availability of pure enzyme should aid in the detection of radiation induced alterations which may be involved in the mechanism of its release from the cell wall.


Enhanced dehydrogenation properties of MgH2by the synergetic effects of transition metal carbides and graphene.
Transition metal carbides show remarkable catalysis for MgH2, and the addition of carbon materials can attach excellent cycling stability. In this paper, Mg-doped with transition metal carbides (TiC) and graphene (G) composite (denoted as Mg-TiC-G) is designed to assess the influence of TiC and graphene on the hydrogen storage performance of MgH2. The as-prepared Mg-TiC-G samples showed favorable dehydrogenation kinetics compared to the pristine Mg system. After adding TiC and graphene, the dehydrogenation activation energy of MgH2decreases from 128.4 to 111.2 kJ mol-1. The peak desorption temperature of MgH2doped with TiC and graphene is 326.5 °C, which is 26.3 °C lower than the pure Mg. The improved dehydrogenation performance of Mg-TiC-G composites is attributed to synergistic effects between catalysis and confinement.


Kinetic mechanism for p38 MAP kinase alpha. A partial rapid-equilibrium random-order ternary-complex mechanism for the phosphorylation of a protein substrate.
p38 Mitogen-activated protein kinase alpha (p38 MAPKalpha) is a member of the MAPK family. It is activated by cellular stresses and has a number of cellular substrates whose coordinated regulation mediates inflammatory responses. In addition, it is a useful anti-inflammatory drug target that has a high specificity for Ser-Pro or Thr-Pro motifs in proteins and contains a number of transcription factors as well as protein kinases in its catalog of known substrates. Fundamental to signal transduction research is the understanding of the kinetic mechanisms of protein kinases and other protein modifying enzymes. To achieve this end, because peptides often make only a subset of the full range of interactions made by proteins, protein substrates must be utilized to fully elucidate kinetic mechanisms. We show using an untagged highly active form of p38 MAPKalpha, expressed and purified from Escherichia coli[Szafranska AE, Luo X & Dalby KN (2005) Anal Biochem336, 1-10) that at pH 7.5, 10 mm Mg2+ and 27 degrees C p38 MAPKalpha phosphorylates ATF2Delta115 through a partial rapid-equilibrium random-order ternary-complex mechanism. This mechanism is supported by a combination of steady-state substrate and inhibition kinetics, as well as microcalorimetry and published structural studies. The steady-state kinetic experiments suggest that magnesium adenosine triphosphate (MgATP), adenylyl (beta,gamma-methylene) diphosphonic acid (MgAMP-PCP) and magnesium adenosine diphosphate (MgADP) bind p38 MAPKalpha with dissociation constants of KA = 360 microm, KI = 240 microm, and KI > 2000 microm, respectively. Calorimetry experiments suggest that MgAMP-PCP and MgADP bind the p38 MAPKalpha-ATF2Delta115 binary complex slightly more tightly than they do the free enzyme, with a dissociation constant of Kd approximately 70 microm. Interestingly, MgAMP-PCP exhibits a mixed inhibition pattern with respect to ATF2Delta115, whereas MgADP exhibits an uncompetitive-like pattern. This discrepancy occurs because MgADP, unlike MgAMP-PCP, binds the free enzyme weakly. Intriguingly, no inhibition by 2 mm adenine or 2 mm MgAMP was detected, suggesting that the presence of a beta-phosphate is essential for significant binding of an ATP analog to the enzyme. Surprisingly, we found that inhibition by the well-known p38 MAPKalpha inhibitor SB 203580 does not follow classical linear inhibition kinetics at concentrations > 100 nm, as previously suggested, demonstrating that caution must be used when interpreting kinetic experiments using this inhibitor.


EFFICACY OF AIR TAMPONADE TREATMENT OF IDIOPATHIC MACULAR HOLES OF DIFFERENT DIAMETERS AND OF FOLLOW-UP INTRAVITREAL AIR TAMPONADE FOR PERSISTENT HOLES.
To evaluate the efficacy of air tamponade in idiopathic macular hole (iMH) surgery and of an additional intravitreal air injection in the treatment of persistent holes. Retrospective, observational case series. Sixty eyes of 60 patients with an iMH underwent phacoemulsification of cataract (when appropriate), pars plana vitrectomy, and internal limiting membrane peeling, followed by air tamponade. Eyes with persistent holes underwent an additional intravitreal air injection within 1 week after surgery. The iMH closure rate and the best-corrected visual acuity were evaluated. In all 30 eyes with an iMH diameter <400 µm, the iMH closed after the primary surgery; however, only 17 of 30 eyes with an iMH diameter of ≥400 µm closed after the primary surgery. For the 13 eyes with persistent holes, an additional intravitreal air injection resulted in successful hole closure. There was no significant difference in the best-corrected visual acuity at the final follow-up between the closed subgroup and the initially unclosed subgroup after closure. Pars plana vitrectomy combined with air tamponade effectively cured small iMHs. For large iMHs not closed after the primary surgery, an additional intravitreal air injection resulted in hole closure and achieved a good prognosis.


Atelocollagen-mediated local and systemic applications of myostatin-targeting siRNA increase skeletal muscle mass.
RNA interference (RNAi) offers a novel therapeutic strategy based on the highly specific and efficient silencing of a target gene. Since it relies on small interfering RNAs (siRNAs), a major issue is the delivery of therapeutically active siRNAs into the target tissue/target cells in vivo. For safety reasons, strategies based on vector delivery may be of only limited clinical use. The more desirable approach is to directly apply active siRNAs in vivo. Here, we report the effectiveness of in vivo siRNA delivery into skeletal muscles of normal or diseased mice through nanoparticle formation of chemically unmodified siRNAs with atelocollagen (ATCOL). ATCOL-mediated local application of siRNA targeting myostatin, a negative regulator of skeletal muscle growth, in mouse skeletal muscles or intravenously, caused a marked increase in the muscle mass within a few weeks after application. These results imply that ATCOL-mediated application of siRNAs is a powerful tool for future therapeutic use for diseases including muscular atrophy.


Determination of clinically therapeutic endoxifen concentrations based on efficacy from human MCF7 breast cancer xenografts.
Tamoxifen is a widely prescribed adjuvant anti-estrogen agent for estrogen receptor-positive breast cancer. Tamoxifen is known to undergo CYP2D6-mediated bioactivation to the active metabolite endoxifen. Endoxifen concentrations exhibit high interindividual variability, contributing to either sub-optimal tamoxifen efficacy or side effects in subsets of patients. However, the relationship between endoxifen exposure and tumor growth inhibition has not been well-characterized and little is known regarding the optimal in vivo endoxifen plasma level required for tumor inhibition. Pharmacokinetics-Pharmacodynamics (PK-PD) modeling was carried out to characterize the relationship between endoxifen concentration and tumor growth inhibition (TGI) in dose-ranging experiments in the human MCF7 xenograft bearing mouse model. Subsequently, simulations using human PK were used to determine the efficacious clinically relevant endoxifen concentration required to produce optimal tumor suppression. Based on the PK-PD model and simulations using clinical PK/concentration data of endoxifen, C stasis (100 % TGI) is observed at 53 nM, a concentration attained by many tamoxifen-treated patients. Importantly, PK-PD simulations indicate that mean steady-state levels observed in CYP2D6 extensive metabolizers are expected to result in optimal tumor suppression while mean concentrations observed in poor metabolizers are predicted to result in suboptimal TGI. Our study is the first to characterize the in vivo PK-PD relationship for endoxifen where clinically observed endoxifen concentrations are associated, in an exposure-dependent manner, with % TGI measured in a xenograft model. It is anticipated that endoxifen concentration achieved in individual patients is the limiting factor for achieving optimal tumor growth suppression.


The heme-binding protein HbpS regulates the activity of the Streptomyces reticuli iron-sensing histidine kinase SenS in a redox-dependent manner.
The SenS/SenR system of Streptomyces reticuli regulates the expression of the redox regulator FurS, the catalase-peroxidase CpeB and the heme-binding protein HbpS. SenS/SenR is also proposed to participate in sensing redox changes, mediated by HbpS. Here, we show in vitro that heme-free HbpS represses the autokinase activity of SenS; whereas hemin-treated HbpS considerably enhances SenS autophosphorylation under redox conditions using either H(2)O(2) or DTT. The presence of iron ions alone or in combination with H(2)O(2) or DTT also leads to significantly increased phosphorylation levels of SenS. Further comparative physiological studies using the S. reticuli WT, a S. reticuli hbpS mutant and a S. reticuli senS-senR mutant corroborates the importance of HbpS and the SenS/SenR system for resistance against high concentrations of iron ions and hemin in vivo. Hence SenS/SenR and HbpS act in concert as a novel three-component system which detects redox stress, mediated by iron ions and heme.


An evaluation of the deterrent impact of Ontario's 12-hour licence suspension law.
The deterrent impact of Ontario's 12-hour licence suspension law, a law intended to mete out swift punishment is evaluated. A process model of deterrence is included. Time series analyses of the monthly log odds of number of positive B.A.C. fatalities over the number of negative B.A.C. fatalities indicated a small, short-term effect. The intermediate measures of four surveys of media coverage, Toronto residents, Ontario residents, and police forces supported these results, and affirmed the potential validity of the process model of deterrence. The results suggest that laws to increase the celerity and certainty of punishment will have little deterrent impact without enforcement and publicity of the new laws.


Voltage-clamp analysis of nodes of Ranvier in regenerated rat sciatic nerve.
Crush-lesioned adult rat sciatic nerves were allowed to regenerate during 70-181 days. Single large regenerated nerve fibres were isolated from levels distal to the crush site, and mounted in a feedback system for potential recordings and voltage clamp. After the experiment, each isolated fibre was fixed and embedded for morphological evaluation. Internodal lengths and fibre diameters were obtained form teased preparations of regenerated and normal sciatic nerves. The isolated regenerated fibres were excitable and had action potentials of large amplitude. TEA and 4-AP had negligible effects on the action potential and excitability properties. The time constant of the nodal segment was larger than in normal fibres. The nodal Na and K permeabilities (PNa and PK) were calculated both in absolute values and relative to the leak conductance (gL). Regenerated fibres had normal PNa/gL and PK/gL ratios. The potential dependence of PNa activation and inactivation was also normal. The isolated physiologically examined fibres had internodal lengths (L) of about 300 micron and diameters (D) of about 7-11 micron. In the teased fibre preparations L was about 300 micron (range 150-600 micron) and D was 3-11 micron. In addition, all teased regenerated nerve preparations exhibited a few scattered unusually short internodes (L less than 150 micron). In control nerves L ranged from 150 micron in the lower fibre size range (D = 2-3 micron) to 1.6 mm in the upper size range (D = 15 micron).


Synchronous oscillations of cytoplasmic Ca2+ and insulin release in glucose-stimulated pancreatic islets.
The cytoplasmic Ca2+ concentration ([Ca2+]i) was measured in single pancreatic mouse islets superfused in a system allowing concomitant recordings of insulin release. When glucose was raised from 3 to 11 mM, [Ca2+]i responded by a transient lowering followed by a rise to an average level of 192 +/- 11 nM. In 77% of the islets the rise was associated with the gradual appearance of oscillations, which were either fast (2-7/min), slow (0.3-0.9/min), or a combination of both types. The characteristics of the fast [Ca2+]i oscillations were those expected from a relationship with the electrical burst activity in islets. Accordingly, in most cases the fast oscillations were remarkably regular. The slow [Ca2+]i oscillations had characteristics similar to the large amplitude ones in individual beta-cells. Whereas glucagon and dibutyryl cAMP could transform slow islet oscillations into fast ones, the alpha 2-adrenergic agonist clonidine had the opposite effect. The rapid islet oscillations were also facilitated by elevated concentrations of extracellular Ca2+. Reinforcing the arguments for [Ca2+]i oscillations as responsible for a pulsatile insulin secretion it was possible to demonstrate that the release of the hormone from single islets is synchronized with the slow [Ca2+]i oscillations.


Chemotactic factor induced tyrosine phosphorylation of membrane associated proteins in rabbit peritoneal neutrophils.
Protein tyrosine phosphorylation in rabbit peritoneal neutrophils was examined by immunoblotting with antibodies specific for phosphotyrosine. Two tyrosine phosphorylated proteins were found with apparent molecular weights of 62,000 (p62) and 125,000. Both were enriched in the membrane fraction. Stimulation of the neutrophils with chemotactic factor fMet-Leu-Phe (10(-8)M, 20 sec) but not phorbol 12-myristate-13-acetate (0.1 microgram/ml, 10 min) caused rapid increase in tyrosine phosphorylation. The effect of fMet-Leu-Phe was inhibited by the pretreatment of neutrophils with pertussis toxin. The p62 protein was also recognized by antibody raised against a synthetic fragment commonly found in the tyrosine kinases of the src gene family. The results indicate that stimulation of the tyrosine phosphorylation of membrane associated proteins is one of the early events occurring in activated neutrophil and this stimulation of tyrosine phosphorylation may be regulated by a GTP-binding protein.


Neonatal lupus erythematosus manifested as a complete heart block: A case report.
Fetal bradycardia and congenital complete heart block could be the presentation of neonatal lupus. A high index of suspicion of this condition helps to identify an asymptomatic mother. Neonatal lupus erythematosus (NLE) is a rare acquired autoimmune disease of newborns due to placental transfer of Ro/SSA or La/SSB autoantibodies. Though cardiac, cutaneous, hematological, and hepatobiliary abnormalities are detected, cardiac defects cause significant morbidity and mortality. We report a case of complete congenital heart block due to NLE.


Emergence and spreading of the largest SARS-CoV-2 deletion in the Delta AY.20 lineage from Uruguay.
The genetic variability of SARS-CoV-2 (genus Betacoronavirus, family Coronaviridae) has been scrutinized since its first detection in December 2019. Although the role of structural variants, particularly deletions, in virus evolution is little explored, these genome changes are extremely frequent. They are associated with relevant processes, including immune escape and attenuation. Deletions commonly occur in accessory ORFs and might even lead to the complete loss of one or more ORFs. This scenario poses an interesting question about the origin and spreading of extreme structural rearrangements that persist without compromising virus viability. Here, we analyze the genome of SARS-CoV-2 in late 2021 in Uruguay and identify a Delta lineage (AY.20) that experienced a large deletion (872 nucleotides according to the reference Wuhan strain) that removes the 7a, 7b, and 8 ORFs. Deleted viruses coexist with wild-type (without deletion) AY.20 and AY.43 strains. The Uruguayan deletion is like those identified in Delta strains from Poland and Japan but occurs in a different Delta clade. Besides providing proof of the circulation of this large deletion in America, we infer that the 872-deletion arises by the consecutive occurrence of a 6-nucleotide deletion, characteristic of delta strains, and an 866-nucleotide deletion that arose independently in the AY.20 Uruguayan lineage. The largest deletion occurs adjacent to transcription regulatory sequences needed to synthesize the nested set of subgenomic mRNAs that serve as templates for transcription. Our findings support the role of transcription sequences as a hotspot for copy-choice recombination and highlight the remarkable dynamic of SARS-CoV-2 genomes.


TOC1 clock protein phosphorylation controls complex formation with NF-YB/C to repress hypocotyl growth.
Plant photoperiodic growth is coordinated by interactions between circadian clock and light signaling networks. How post-translational modifications of clock proteins affect these interactions to mediate rhythmic growth remains unclear. Here, we identify five phosphorylation sites in the Arabidopsis core clock protein TIMING OF CAB EXPRESSION 1 (TOC1) which when mutated to alanine eliminate detectable phosphorylation. The TOC1 phospho-mutant fails to fully rescue the clock, growth, and flowering phenotypes of the toc1 mutant. Further, the TOC1 phospho-mutant shows advanced phase, a faster degradation rate, reduced interactions with PHYTOCHROME-INTERACTING FACTOR 3 (PIF3) and HISTONE DEACETYLASE 15 (HDA15), and poor binding at pre-dawn hypocotyl growth-related genes (PHGs), leading to a net de-repression of hypocotyl growth. NUCLEAR FACTOR Y subunits B and C (NF-YB/C) stabilize TOC1 at target promoters, and this novel trimeric complex (NF-TOC1) acts as a transcriptional co-repressor with HDA15 to inhibit PIF-mediated hypocotyl elongation. Collectively, we identify a molecular mechanism suggesting how phosphorylation of TOC1 alters its phase, stability, and physical interactions with co-regulators to precisely phase PHG expression to control photoperiodic hypocotyl growth.


Development and validation of a peripheral cell ratio and lactate score for differentiating status epilepticus from prolonged psychogenic nonepileptic seizures.
Differentiating status epilepticus (SE) from prolonged psychogenic nonepileptic seizures (pPNES) can be difficult clinically. We aimed to define the utility of peripheral cell counts, cell ratios, and lactate levels in distinguishing SE from pPNES. Retrospective two-center study investigating the sensitivity and specificity of acute (≤12 h of event offset) peripheral cell counts, cell ratios (neutrophil-lymphocyte ratio, neutrophil-monocyte ratio, monocyte-lymphocyte ratio, platelet-lymphocyte ratio, systemic immune-inflammatory index [SII], systemic inflammatory response index [SIRI]), and lactate levels in differentiating SE from pPNES. Patients were identified from two tertiary hospitals, with one forming the development cohort and the other the validation cohort. Using generalized additive models to generate biomarker vs time curves, optimal blood collection times were defined for set parameters. Three diagnostic scores combining neutrophil count, SII, or SIRI with lactate levels were developed and validated in separate cohorts. For the development cohort, 1262 seizure-like events were reviewed and 79 SE and 44 pPNES events were included. For the validation cohort, 241 events were reviewed and 20 SE and 11 pPNES events were included. Individually, the biomarkers generally had low sensitivity and reasonable specificity for differentiating SE from pPNES, with the neutrophil count, SIRI, and SII performing best with sensitivities of 0.65-0.84, specificities of 0.64-0.89, and ROC AUCs of 0.78-0.79. Lactate levels peaked at 60 min, while cell counts and ratios peaked after 240 min. Combining early peaking lactate levels and later peaking neutrophil count, SIRI or SII resulted in three scores that improved predictive potential with sensitivities of between 0.75 and 0.79, specificities between 0.93 and 1.00, and ROC AUCs of 0.89-0.91. Lactate levels peak early post-SE, whereas cell counts and ratios do so later. The differing post-event time profiles of lactate levels vs neutrophil count, SIRI, and SII allow incorporation into three separate scores which can assist in differentiating SE from pPNES.


Glutathione-S-transferase P1, T1 and M1 genetic polymorphisms in neoadjuvant-treated locally advanced gastric cancer: GSTM1-present genotype is associated with better prognosis in completely resected patients.
Neoadjuvant chemotherapy in gastric cancer is now standard in the Western world; however, only 30-40% of the patients respond to induction therapy. Pretherapeutic predictors of response and prognosis would be of utmost interest to individualize treatment. Glutathione-S-transferase enzymes detoxify therapeutic drugs such as platin derivates and may influence outcome of the treated patients. Therefore, glutathione-S-transferase (GST) polymorphisms were assessed as predictive markers in cisplatinum-based neoadjuvant-treated gastric cancer. DNA was isolated from 139 patients with locally advanced gastric cancer (cT3/4 anyN cM0) before chemotherapy. Multiplex polymerase chain reaction was used for GSTT1 and GSTM1 genes, and allelic discrimination assay with the TaqMan system for the GSTP1 gene. One hundred ten patients could be analyzed for GSTT1 (T-:23; T+87), 112 for GSTM1 (M-:52; M+:60) and 132 for GSTP1 (Ile/Ile: 55; Ile/Val: 59; Val/Val: 18). There was no significant correlation between any of the GSTT1, GSTM1, or GSTP1 genotypes and patients' characteristics or histopathological data; only the GSTM1+ genotype was associated with the non-intestinal subtype of the Lauren classification (p=0.045). GSTT1, GSTM1, and GSTP1 genotypes were not correlated with response to chemotherapy (p=0.57, p=0.38, p=0.33). In R0 resected patients, we found an improved survival for patients with the GSTM1-present genotype compared to patients with the GSTM1-null genotype (p=0.017). Moreover, the GSTM1-present genotype showed a significantly better tumor-related (p=0.017) and disease-free survival (p=0.029). None of the common GST polymorphisms predicts response in our study, but the GSTM1+ genotype was associated with a better prognosis in completely resected patients. Further investigations on chemotherapy-associated gene polymorphisms are warranted.


Hydronephrosis with ureteral valve: diagnosis by ultrasonography and antegrade pyelography.
A case of massive hydronephrosis secondary to a ureteral valve was discovered upon evaluation of mild trauma. The orderly diagnostic studies, including ultrasonography and antegrade pyelography, led to the correct diagnosis and appropriate therapy. Ultrasonography as a diagnostic tool in urology is discussed and characteristics of the 18 cases of ureteral valves reported to date are reviewed.


Geographic isolation affects inter- and intra-specific seed variability in the Astragalus tragacantha complex, as assessed by morpho-colorimetric analysis.
The effect of geographical isolation on the inter- and intra-specific seed variability of the Astragalus tragacantha complex was investigated by using a computer-aided imaging system. In particular, seed morphometric and colorimetric features of Astragalus balearicus, A. tegulensis, A. terraccianoi, A. thermensis and A. tragacantha were measured and the data were used to discriminate among regions of provenance, taxa and populations. Discriminant analysis of the data supports the accepted taxonomy of the group and the hypothesis that interspecific differentiation resulted from geographical isolation. The results also showed a positive correlation between the wideness of species' distribution and the levels of seed intra-specific variability.


Incidental pulmonary agenesis with multiple associated anomalies: A case report.
Pulmonary agenesis is an uncommon congenital abnormality of the lungs that occurs either in isolation or in association with other anomalies in several organ systems. Right sided pulmonary agenesis is reported to have a worse prognosis owing to the more severe mediastinal shift and resultant compression of mediastinal structures. We present a case of right-side pulmonary agenesis with multiple associated anomalies found incidentally in a 19-year-old female patient.


Mandibular lateral deviation induces alteration in vascular endothelial growth factor expression and oxidative stress/nitric oxide generation in rat condyle, synovial membrane and masseter muscle.
We aimed to investigate alteration in cellular signaling mediated by vascular endothelial growth factor (VEGF) and parameters of oxidative stress/nitric oxide generation, superoxide dismutase (SOD) and neuronal nitric oxide synthase (nNOS), underlying altered functional mechanical loading of TMJ (temporomandibular joint) during lateral mandibular deviation. Thirty-eight 5-week-old male Wistar rats were divided into experimental group, which received acrylic resin appliance that shifted mandible to the left during closure, and control group. Computed tomography and histomorphometry were used for condyle analyses, while samples of condyle, synovial membrane and m. masseter were analyzed with enzyme-linked immunosorbent assay and spectrophotometry to determine VEGF and nNOS protein concentrations, and SOD activity. Experimental group of rats developed smaller and asymmetrical mandibles. Less of new bone and cartilage formation and larger bone marrow cavities area were found in the experimental group. Higher VEGF expression in condyle and m. masseter as well as higher nNOS expression in m. masseter and synovial membrane were found in the experimental compared to the control group. Alteration of SOD activity was found in m. masseter and synovial membrane in the experimental group. Lateral mandibular deviation induces mandibular and condylar morphological changes as well as significant cellular signaling alterations in condyle, synovial membrane and masticatory muscle. Cellular VEGF protein overexpression and oxidative stress/nitric oxide disbalance could be the mechanisms underlying unbalanced functional TMJ loading due to mandibular deviation.


Epidermal growth factor induces cytokeratin 19 expression accompanied by increased growth abilities in human hepatocellular carcinoma.
Cytokeratin (CK) 19-positive hepatocellular carcinoma (HCC) has been reported to have a poor prognosis. The mechanism of the development of CK19-positive HCC remains to be studied. To clarify this, in vitro experiments were performed using human HCC cell lines (PLC-5, HepG2), and the phenotypic changes after stimulation with several growth factors were examined using quantitative reverse transcriptase PCR, western blotting, and immunofluorescence staining. In vivo experiments using human HCC specimens obtained from a total of 78 patients and clinicopathological analysis were also performed. Among the growth factors tested, epidermal growth factor (EGF) had prominent effects on inducing CK19 expression in PLC-5 and HepG2, which was accompanied by the reduced expression of α-fetoprotein in PLC-5. The induction of CK19 expression after EGF stimulation was accompanied by the phosphorylation of c-Jun-N-terminal kinase (JNK)/stress-activated protein kinase, which was blocked by the addition of JNK inhibitors. EGF also increased proliferative abilities and invasive properties of the HCC cell lines. In vivo, 9 (12%) of 78 HCC cases showed positive immunohistochemical staining of CK19. The extent of positive immunohistochemical signals of EGF, EGF receptor (EGFR), and JNK expression was significantly intense in CK-19-positive HCC than those of CK19-negative HCC. Clinicopathological analysis showed that CK19-positive HCC had a high incidence of portal vein invasion, extrahepatic metastasis and an early relapse, which was associated with the worsened 2-year disease free survival. These results indicate that the activation of the EGF-EGFR signaling pathway is associated with the development of CK19-positive HCC, and the EGF-induced increase in growth abilities of HCC may account for the poor prognosis of the patients.