JNK pathway may be involved in isoflurane-induced apoptosis in the hippocampi of neonatal rats.
Previous studies have demonstrated that isoflurane, a commonly used volatile anesthetic, can induce widespread apoptosis in the neonatal animal brains and result in persistent cognitive impairment. Isoflurane-induced cytosolic Ca(2+) overload and activation of mitochondrial pathway of apoptosis may be involved in this neurodegeneration. The c-Jun N-terminal kinase (JNK) signaling can regulate the expression of the Bcl-2 family members that modulates mitochondrial membrane integrity. Therefore, we hypothesize that JNK signaling pathway activation contributes to isoflurane-induced apoptosis in the brain. In this study, Sprague-Dawley neonatal rats at postnatal day 7 were exposed to 1.1% isoflurane or air for 4h. The JNK inhibitor SP600125 at 5 μg, 10 μg, 20 μg, 30 μg or the vehicle was intraventricularly administered before the exposure. Neuronal apoptosis in the hippocampi of neonatal rats was detected by TUNEL 6h after isoflurane or air exposure. The protein expression of phospho-JNK, phospho-c-Jun, and caspase-3 as well as the antiapoptotic protein Bcl-xL and Akt/glycogen synthase kinase (GSK)-3β pathway was detected by Western blotting. Isoflurane significantly increased apoptotic cells in the hippocampal CA1, CA3, and DG regions. The JNK inhibitor SP600125 dose-dependently inhibited isoflurane-induced neuronal apoptosis and increase of caspase-3 and phospho-JNK. SP600125 also attenuated isoflurane-induced down-regulation of Bcl-xL and maintained the activated Akt level to increase the phosphorylation of GSK-3β at Ser9. Our results indicate that JNK activation contributes to isoflurane-induced neuroapoptosis in the developing brain. Maintaining Bcl-xL and Akt activation may be involved in the neuroprotective effects of SP600125.


The GAMYB-like gene SlMYB33 mediates flowering and pollen development in tomato.
GAMYBs are positive GA signaling factors that exhibit essential functions in reproductive development, particularly in anther and pollen development. However, there is no direct evidence of the regulation of any GAMYB in these biological processes in tomato (Solanum lycopersicum). Here, we identified a tomato GAMYB-like gene, SlMYB33, and characterized its specific roles. SlMYB33 is predominately expressed in the stamens and pistils. During flower development, high mRNA abundance of SlMYB33 is detected in both male and female organs, such as microspore mother cells, anthers, pollen grains, and ovules. Silencing of SlMYB33 leads to delayed flowering, aberrant pollen viability, and poor fertility in tomato. Histological analyses indicate that SlMYB33 exerts its function in pollen development in the mature stage. Further transcriptomic analyses imply that the knockdown of SlMYB33 significantly inhibits the expression of genes related to flowering in shoot apices, and alters the transcription of genes controlling sugar metabolism in anthers. Taken together, our study suggests that SlMYB33 regulates tomato flowering and pollen maturity, probably by modulating the expression of genes responsible for flowering and sugar metabolism, respectively.


SAFE-clustering: Single-cell Aggregated (from Ensemble) clustering for single-cell RNA-seq data.
Accurately clustering cell types from a mass of heterogeneous cells is a crucial first step for the analysis of single-cell RNA-seq (scRNA-Seq) data. Although several methods have been recently developed, they utilize different characteristics of data and yield varying results in terms of both the number of clusters and actual cluster assignments. Here, we present SAFE-clustering, single-cell aggregated (From Ensemble) clustering, a flexible, accurate and robust method for clustering scRNA-Seq data. SAFE-clustering takes as input, results from multiple clustering methods, to build one consensus solution. SAFE-clustering currently embeds four state-of-the-art methods, SC3, CIDR, Seurat and t-SNE + k-means; and ensembles solutions from these four methods using three hypergraph-based partitioning algorithms. Extensive assessment across 12 datasets with the number of clusters ranging from 3 to 14, and the number of single cells ranging from 49 to 32, 695 showcases the advantages of SAFE-clustering in terms of both cluster number (18.2-58.1% reduction in absolute deviation to the truth) and cluster assignment (on average 36.0% improvement, and up to 18.5% over the best of the four methods, measured by adjusted rand index). Moreover, SAFE-clustering is computationally efficient to accommodate large datasets, taking <10 min to process 28 733 cells. SAFEclustering, including source codes and tutorial, is freely available at https://github.com/yycunc/SAFEclustering. Supplementary data are available at Bioinformatics online.


Anti-human platelet antigen-5b antibodies and fetal and neonatal alloimmune thrombocytopenia; incidental association or cause and effect?
Most cases of fetal and neonatal thrombocytopenia (FNAIT) are caused by maternal anti-human platelet antigen-1a antibodies (anti-HPA-1a). Anti-HPA-5b antibodies are the second most common antibodies in suspected FNAIT cases. Given the high prevalence of anti-HPA-5b antibodies in pregnant women delivering healthy newborns, the association with FNAIT may be coincidental. This review of the literature related to FNAIT using the MEDLINE database was conducted according to PRISMA guidelines. A retrospective analysis of a single-centre cohort of 817 suspected FNAIT cases was conducted. The pooled prevalence of anti-HPA-5b antibodies in unselected pregnant women of European descent was 1.96% (n = 3113), compared with 3.4% (n = 5003) in women with suspected FNAIT. We found weak evidence that a small proportion of pregnant women presenting with anti-HPA-5b antibodies will give birth to a newborn with mild thrombocytopenia. The neonatal platelet counts were not different between suspected FNAIT cases (n = 817) with and without maternal anti-HPA-5b antibodies. The prevalence of maternal anti-HPA-5b antibodies was not different between neonates with intracranial haemorrhage and healthy controls. The current experimental and epidemiological evidence does not support the hypothesis that anti-HPA-5b antibodies cause severe thrombocytopenia or bleeding complications in the fetus or newborn.


Prediction of acute hypotensive episodes by means of neural network multi-models.
This work proposes the application of neural network multi-models to the prediction of adverse acute hypotensive episodes (AHE) occurring in intensive care units (ICU). A generic methodology consisting of two phases is considered. In the first phase, a correlation analysis between the current blood pressure time signal and a collection of historical blood pressure templates is carried out. From this procedure the most similar signals are determined and the respective prediction neural models, previously trained, selected. Then, in a second phase, the multi-model structure is employed to predict the future evolution of current blood pressure signal, enabling to detect the occurrence of an AHE. The effectiveness of the methodology was validated in the context of the 10th PhysioNet/Computers in Cardiology Challenge-Predicting Acute Hypotensive Episodes, applied to a specific set of blood pressure signals, available in MIMIC-II database. A correct prediction of 10 out of 10 AHE for event 1 and of 37 out of 40 AHE for event 2 was achieved, corresponding to the best results of all entries in the two events of the challenge. The generalization capabilities of the strategy was confirmed by applying it to an extended dataset of blood pressure signals, also collected from the MIMIC-II database. A total of 2344 examples, selected from 311 blood pressure signals were tested, enabling to obtain a global sensitivity of 82.8% and a global specificity of 78.4%.


Recombinant Fluorescent Rabies Virus Vectors for Tracing Neurons and Synaptic Connections.
Recombinant rabies virus (RV) vectors expressing fluorescent proteins allow staining of neurons from many mammalian species and enable the study of neuron morphology. Because viral spread occurs only between neurons that have synaptic connections, these vectors also permit transsynaptic tracing. A recently established system for restriction of transsynaptic tracing to a single transsynaptic jump, dubbed monosynaptic tracing, uses glycoprotein gene-defective, pseudotyped RV. This allows infection of defined cells and transient complementation with the glycoprotein in situ to support a single step of transsynaptic crossing to presynaptic cells. Here, we introduce protocols describing the production of RV vectors, including the recovery of recombinant RV from complementary DNA (cDNA) and virus pseudotyping in vitro. This allows retrograde staining of neurons projecting to the inoculation site.


Enhancement of colony-stimulating-factor--dependent clonal growth of murine macrophage progenitors and their phagocytic activity by retinoic acid.
The effect of retinoic acid (RA) on the colony-stimulating-factor-dependent clonal growth of myeloid progenitors was assessed in semisolid agar cultures of mouse bone marrow cells using L-cell-conditioned medium that gave rise to macrophage colonies, granulocyte colonies, and mixed macrophage-granulocyte colonies and clusters. RA was found to enhance the overall formation of myeloid colonies (about 50%) and clusters in 7-day cultures. The increase was due to an enhanced formation of macrophage colonies (70-250%) and clusters which reached a maximal value at about 3 microM RA. In 4-day cultures, the effect of RA on macrophage colony formation was biphasic with a maximal enhancement at 10 nM. RA suppressed granulocyte-colony formation in 4-day cultures. RA increased the phagocytic activity of bone-marrow-derived macrophages at all stages of differentiation and/or maturation in culture. The Fc-receptor-mediated erythrophagocytosis as well as the phagocytosis of heat-killed yeast cells (HK-yeast) and starch particles increased by RA treatment in a dose-dependent manner, reaching an increase of 100-200% of the activity expressed in the absence of RA. Peritoneal exudate macrophages likewise exhibited an increased phagocytic response to a variety of particles, at both physiological and pharmacological concentrations of RA. Expression of an RA-mediated increase in phagocytic activity required a prolonged incubation with RA (greater than 19 hr). The data suggest that RA may be of physiological relevance in the regulation of proliferation and function of hemopoietic cells. Therapeutic doses of RA may potentiate macrophage proliferation and function, elements that are crucial at all phases of the various defense mechanisms that the organism possesses.


Oxidative stress in neonatal lambs and its relation to health status and passive colostral immunity.
Oxidative stress (OS) related to birth in ruminants may increase susceptibility to diseases. However, information about the physiological changes related to OS in lambs is limited. This study was designed to evaluate changes in OS parameters [total antioxidant status (TAS, mmol Trolox Equiv./L), total oxidant status (TOS, μmol H2O2 Equiv./L)] and calculated oxidative stress index (OSi = TOS/TAS * 100, Arbitrary Unit) and its association with passive colostral transfer immunity and health status in neonatal Akkaraman crossbreed lambs. Control group of healthy lambs (n = 15) were blood sampled before (day 0) and after colostrum intake (on day 1, 2, 4, 7, 14 and 28) in order to evaluate the effect of colostrum intake on TAS, TOS and OSi over time. In addition, a group of lambs (experimental group, n = 60), based on their health status and different cut off value of IgG (mg/dL), was also blood sampled at 24 ± 1 h of colostrum ingestion to determine the relationship between OS parameters, IgG and health status. OS was evident at birth and gradually decreased in control group as TOS (days 0:8.06 ± 0.64, 1:7.05 ± 0.51, 2:5.23 ± 0.35, 4:5.05 ± 0.2, 7:5.13 ± 0.34, 14:3.75 ± 0.12 and 28:3.73 ± 0.15 μmol H2O2 Equiv./L), OSi (days 0: 8.35 ± 0.64, 1: 9.17 ± 1.05, 2:6.83 ± 0.6, 4:6.30 ± 0.20, 7:6.67 ± 0.59, 14:4.46 ± 0.19 and 28:4.87 ± 0.29 au) and TAS (days 0: 0.97 ± 0.02, 1:0.82 ± 0.04, 2:0.81 ± 0.05, 4:0.81 ± 0.02, 7:0.80 ± 0.03, 14:0.85 ± 0.02 and 28:0.79 ± 0.03 mmol Trolox Equiv./L) values changed over time. In the experimental group, the lambs died in the first week of the neonatal period had higher levels of TOS (10.3 ± 0.90 μmol H2O2 Equiv./L) and OSi (1.61 ± 0.22 au) than healthy ones (TOS:6.90 ± 0.36 μmol H2O2 Equiv./L, OSi:1.12 ± 0.11 au) (P < 0.001). The gradual decrease in OS in the neonatal period was accepted as a sign of improved antioxidant response. The study disclosed that deaths may develop when OS at birth is not balanced by suffıcient amount and quality of colostrum an important source of antioxidants such as enzymes and vitamins. The study presented may provide new information in the field of study on oxidative stress in neonatal lambs and help to prevent deaths in the first week of life. It can also be useful in developing preventive measures such as supplements of antioxidants to colostrum.


Mushroom keratoplasty in pediatric patients.
To report the outcome of mushroom keratoplasty for the treatment of full thickness corneal disease in pediatric patients with healthy endothelium. A retrospective analysis of pediatric patients who underwent mushroom keratoplasty. The medical records of pediatric patients suffering from full thickness corneal stromal disease with normal endothelium who underwent mushroom keratoplasty at our Institution were included. A two-piece donor graft consisting of a large anterior stromal lamella (9.0 mm in diameter and ±250 μm in thickness) and a small posterior lamella (5-6.5 mm in diameter) including deep stroma and endothelium, prepared with the aid of a microkeratome had been transplanted in all cases. Ophthalmic examination including slit lamp examination, best corrected visual acuity, and corneal topography was performed preoperatively and at each postoperative visit on all patients. The endothelial cells were assessed by specular microscopy in these patients. Six eyes of six patients (five males and one female) were included. The mean age was 9.3 years (range 5-15 years). Average follow-up was 17.8 months (range 9-48 months). There were no early or late complications recorded. All corneas were clear at the last follow up visit. Preoperative best corrected visual acuity (BCVA) was worse than 20/70 in all six eyes. Postoperatively, four eyes achieved BCVA of 20/40 or better. Endothelial cell loss (n eyes = 3 averaged 24% (range 19-31%). The mean endothelial cell loss was 24% (range 19-31%) among these patients. Microkeratome assisted mushroom keratoplasty is a viable surgical option for pediatric eyes with full thickness corneal stromal disease and healthy endothelium. Mushroom keratoplasty combines the refractive advantage of a large penetrating keratoplasty with the survival advantage of a small penetrating keratoplasty. Furthermore, mushroom keratoplasty exhibits the mechanical advantage of a shaped keratoplasty.


Occupation and risk of cancer of the renal pelvis in Nordic countries.
To evaluate occupational variation in the incidence of the malignant neoplasm of the renal pelvis in the population of the Nordic countries: Denmark, Iceland, Finland, Norway and Sweden. The study cohort comprised 14.9 million individuals. Data on occupational history were obtained from national censuses. Standardized incidence ratios (SIRs) were calculated for each occupation. The highest SIRs were found in seamen (1.51, 95% confidence interval [CI] 1.23-1.82), printers (1.39, 95% CI 1.11-1.71), welders (1.37, 95% CI 1.03-1.78), and public safety workers (1.35, 95% CI 1.12-1.62). The lowest SIRs were observed in forestry workers (0.47, 95% CI 0.35-0.62), gardeners (0.72, 95% CI 0.62-0.83) and woodworkers (0.81, 95% CI 0.72-0.91). The study suggests that there is an association between profession and risk of malignancy of the renal pelvis. The possible associations between exposure to asbestos, heavy metals and welding fumes, and risk of developing the disease should be studied further.


The philosophical bases of grounded theory and their implications for research practice.
What are the implications of using a grounded theory approach in nursing research? Liz Norton discusses the issues and offers some personal insights.


Effect of acetaldehyde on rat platelet aggregation in vivo and in vitro.
The effect of acetaldehyde on primary hemostasis and platelet aggregation was studied in vivo and in vitro in the rat. Acetaldehyde was found in circulation following ethanol intoxication or was present after direct i.v. injection. In vitro, acetaldehyde was added to whole blood or platelet plasma suspension. Bleeding time and blood loss were increased upon ethanol intoxication and immediately after i.v. infusion of acetaldehyde. The intrinsic coagulation system, assayed by APT-time, was unaffected. In vivo, ADP and collagen-induced platelet aggregation was inhibited upon the presence of acetaldehyde. In vitro, inhibition of platelet aggregation was observed when acetaldehyde was added to blood, while the addition of the compound to plasma did not affect platelet function.


Monoclonal antibodies to restricted and cross-reactive idiotopes on monoclonal rheumatoid factors and their recognition of idiotope-positive cells.
Four human monoclonal rheumatoid factors (MRF) were used to raise a panel of mouse monoclonal antibodies (mAb) which were selected in a solid-phase radioimmunoassay for binding to MRF but not normal IgM. Three mAb, each raised against a different MRF, bound to the majority of MRF and also to most polyclonal RF. Four other mAb bound selectively to the MRF against which they were raised and to no other MRF, and rarely to any polyclonal RF. Competition studies using cold and radiolabeled mAb further indicated that these mAb recognize distinct and different epitopes on MRF. RF activity of MRF was inhibited by 3 of the 4 mAb binding to a single MRF and 2 of the 3 mAb binding to multiple RF. It was thus concluded that of this panel of mAb 3 recognized cross-reactive idiotopes and the remainder demonstrated highly restricted idiotopes on MRF. These mAb identified MRF idiotope-bearing cells in the peripheral blood of 3 of the MRF donors (and a further subject with type II essential cryoglobulinemia), with a frequency ranging from 0.3-10% of all mononuclear cells with the mAb to restricted idiotopes or 1.5-17% with mAb to cross-reactive idiotopes. These anti-idiotopic mAb should thus provide a highly specific means of identifying and monitoring MRF-producing cells in vivo.


Two ways to survive infection: what resistance and tolerance can teach us about treating infectious diseases.
A host can evolve two types of defence mechanism to increase its fitness when challenged with a pathogen: resistance and tolerance. Immunology is a well-defined field in which the mechanisms behind resistance to infection are dissected. By contrast, the mechanisms behind the ability to tolerate infections are studied in a less methodical manner. In this Opinion, we provide evidence that animals have specific tolerance mechanisms and discuss their potential clinical impact. It is important to distinguish between these two defence mechanisms because they have different pathological and epidemiological effects. An increased understanding of tolerance to pathogen infection could lead to more efficient treatments for infectious diseases and a better description of host-pathogen interactions.


The added value of CT virtual angioscopy to MDCT angiography in the evaluation of aortic diseases.
Incorporation of virtual angioscopy (VA) in the diagnostic work-up of aortic diseases could improve the clinical value and efficiency of multidetector computed tomography angiography (MDCTA). We aim to evaluate the clinical usefulness of virtual aortic navigation by CT angiography in various aortic diseases as a complement to standard MDCTA. We retrospectively selected 211 patients who performed MDCTA for suspected or operated aortic diseases. VA endoluminal images of the aorta were obtained by a fly-through technique. Two senior vascular radiologists independently evaluated all MDCTA images. After 1 month, the same two radiologists independently reviewed the MDCTA images combined with CTVA images. The respective accuracy of CTVA in delineating aortic abnormalities was compared to that of MDCTA using Fisher's exact test. The Fleiss kappa (κ) statistic was used to assess the inter-reader agreement (IRA). We detected 229 abnormalities in 203 patients on MDCTA and 231 abnormalities in 205 patients on CTVA. CTVA provided significant additional findings in 63.8% (146/229) of all abnormalities diagnosed by MDCTA (p < 0.001, odd ratio [OR] = 42). Although CTVA diagnosed two abnormalities overlooked by MDCTA, the value was statistically insignificant (p = 0.787, OR = 1.3). Regarding postoperative abnormalities, the CTVA added significant additional findings over MDCTA (p = 0.006, OR = 87.4). The overall IRA for the performance of CTVA was good (κ = 0.699). CTVA yields extra findings and improves diagnostic efficiency of MDCTA, especially in postoperative patients.


Comparison of nested PCR and qPCR for the detection and quantitation of BoHV6 DNA.
Nested PCR and qPCR (quantitative PCR) tests based on glycoprotein B (gB) gene were designed for detecting Bovine herpesvirus 6 (BoHV6) in bovine whole blood samples and wild ruminant blood clots (deer and roe-deer). This virus, commonly known as BLHV (bovine lymphotropic herpesvirus) belongs to the Herpesviridae family, subfamily Gammaherpesvirinae and Macavirus genus. DNA isolated from 92 dairy cow blood samples and 69 wild ruminant clots were examined for the presence of BoHV6 using nested PCR and qPCR tests. Viral DNA was detected by using nested PCR in 59 out of 92 bovine blood samples (64.1%), and by qPCR in 68 out of 92 bovine blood samples (73.9%), but none out of 69 DNA samples isolated from wild ruminant blood clots, was positive in both assays. The specificity of nested PCR and qPCR was confirmed by using BoHV1, BoHV4, BoHV6, BFV, BIV, and BLV DNA. The sensitivity of nested PCR and qPCR was determined using a serially 10-fold diluted vector pCR2.1HgB (2 × 10(0)-2 × 10(6)copies/reaction). In this testing, qPCR was more sensitive than the nested PCR, detecting two copies of BoHV6 whilst the limit of detection for nested PCR was 20 copies. In all qPCR assays, the coefficients of determination (R(2)) ranged between 0.990 and 0.999, and the calculated amplification efficiencies (Eff%) within the range of 89.7-106.9. The intra- and inter-assay CV (coefficient of variation) values did not exceed 4%.


Designing Molecular Spanners to Throw in the Protein Networks.
Proteins govern most aspects of cellular life and, through specific interfaces, are typically involved in intricate protein-protein interaction (PPI) networks and signaling pathways. Subtle up- or downregulation of key protein functions and PPIs results in disease; still, the preferred option to contrast the role of a protein in disease and healthy conditions alike remains its outright shutdown through orthosteric ligands that block its active site. Here, we explore subtler alternatives to modulate proteins and PPIs. Driven by a view of proteins as dynamic entities, we discuss ways to identify allosteric binding sites, which, when targeted by tailored ligands, can induce significant changes in the active site of a protein, and lead to agonistic or antagonistic effects. We also summarize the selective regulation of specific PPIs-either direct or allosteric-and show that effects can be stabilizing as well as destabilizing, depending on how the conformational equilibrium of a protein is shifted.


Interleukin-17A is involved in enhancement of tumor progression in murine intestine.
Interleukin (IL)-17A is a cytokine involved in neutrophilic inflammation but the role of IL-17A in anti-tumor immunity is controversial because both pro- and anti-tumor activities of IL-17A have been reported. We hypothesized that constitutive expression of IL-17A in intestinal environment modifies tumor growth. To address the issue, mice were inoculated into subserosa of cecum (i.c.) with murine EL4 lymphoma expressing a model tumor antigen, and tumor growth was monitored. IL-17A-producing cells were detected both in tumor mass and in normal intestinal tissue of i.c. tumor-bearing wild type mice. Tumor size in the wild-type mice was significantly higher than that in the cecum of IL-17A gene-knockout mice. Furthermore, anti-IL-17A monoclonal antibody treatment of wild-type mice resulted in decreased tumor size in the cecum. Model tumor-antigen-specific interferon-γ production was not modified in draining mesenteric lymph node cells in the absence or after neutralization of IL-17A. All the results suggest that constitutive expression of IL-17A in intestine enhances tumor growth, and anti-IL-17A antibody treatment is a candidate of a new anti-tumor immunotherapy against intestinal tumors.


A comparison of clinical outcomes of acute testicular torsion between prepubertal and postpubertal males.
Surgical intervention for acute testicular torsion can require either orchiopexy or orchiectomy. The decision of which surgery to perform is dependant on the amount of time that the testicle experienced ischemia and the viability of the testicle after reperfusion. It is hypothesized that (1) there is a difference in orchiectomy and orchiopexy rates between prepubertal and postpubertal males with acute testicular torsion and (2) presenting symptoms may vary between the two age groups as prepubertal males may present with atypical symptoms, which could result in delayed presentation and diagnosis. A retrospective chart review was conducted on pediatric patients who were diagnosed with acute testicular torsion between June 2010 and August 2017. Demographic and clinical characteristics were extracted: age, ethnicity, referral pattern, primary insurance status, symptoms at presentation, prior history of ipsilateral testicular pain or intermittent torsion, recent trauma to genitalia, duration of symptoms (hours), gradual vs. acute onset of symptoms, time/weekday/season at presentation, and time interval from arrival at the study institution to surgical intervention (minutes). Patients were categorized into two groups: prepubertal group (age 1-12 years) and postpubertal group (age 13-18 years). Statistical analyses were performed using R, version 3.3.1. Ninety-one patients were included in the study. The overall orchiectomy rate was 30.8%. More prepubertal males underwent orchiectomy than postpubertal males (42.4% vs. 24.1%, respectively). Prepubertal males were more likely to present with abdominal pain than postpubertal males (27.3% vs. 10.3%, respectively). Those who underwent orchiectomy were more likely to present with longer duration of symptoms, testicular swelling, and abdominal pain than those who underwent orchiopexy. The risk of orchiectomy decreased by 14% per 1-year increase in age (odds ratio [OR]: 0.86, 95% confidence interval [CI]: 0.94-1.00, p = .009). A steady decline in the proportion of patients undergoing orchiectomy was seen from 1 to 12 years of age. This study found that prepubertal males are at higher risk for orchiectomy than postpubertal males. The risk of orchiectomy decreases by 14-16% per 1-year increase in age. Prepubertal males are more likely to present with atypical symptoms and delayed presentation and diagnosis, leading to delayed surgical intervention. It is important for providers to perform a genital examination in prepubertal males who present with abdominal pain to rule out acute testicular torsion. Patients presenting with longer duration of symptoms, testicular swelling, and abdominal pain are at higher risk for orchiectomy. No correlation was found between orchiectomy rate and ethnicity, referral status, primary insurance status, and time/weekday/season at presentation. Among patients presenting to a tertiary pediatric hospital with acute testicular torsion, prepubertal males (younger than 12 years) are at higher risk for orchiectomy than postpubertal males. Prepubertal males are more likely to present with atypical symptoms which results in delayed presentation and diagnosis, leading to delayed in surgical intervention.


Comparison study of single-port (Octoport™) and four-port total laparoscopic hysterectomy.
To compare the perioperative surgical outcomes and complication rates between single-port total laparoscopic hysterectomy and conventional four-port total laparoscopic hysterectomy. Between June 2009 and April 2011, 56 patients underwent total laparoscopic hysterectomy. Of these 56 patients, 28 underwent single-port (Octoport™) total laparoscopic hysterectomy and 28 underwent conventional four-port total laparoscopic hysterectomy. We analyzed the following parameters for all the patients: age, body mass index, operative time, blood loss, change in hemoglobin level, vaginal stump suture time and length of hospital stay. The general characteristics of the patients were similar in both groups. There were no statistically significant differences in blood loss, hemoglobin change, length of postoperative hospital stay and complication rate. However, the mean operative time of the single-port group was significantly longer than that of the four-port group (93.5±24.0min vs. 78.7±17.4min; P=0.011). The operative time for vaginal stump suture was profoundly decreased with experience in the single-port group. With the exception of operative time, the surgical outcomes and incidence of complications of the single-port group were comparable to those of the four-port group. However, the operative time decreased in the single-port group with increasing experience.


Prevalence of hallucinations in Parkinson's patients: meta-analysis of International studies.
Parkinson's disease (PD) is one of the common causes of disability in the elderly. This study aims to estimate the prevalence of hallucinations in Parkinson's patients in the world. A systematic review of PubMed/Medline, ISI Web of Knowledge, and Google Scholar was conducted from 2017 to 2022. This study investigated the prevalence of hallucinations in Parkinson's patients. Point prevalence was analyzed with a 95% confidence interval. The variances of each study were calculated using the binomial distribution formula The researcher used Cochrane Q-test with a significance level of less than 0.1 to check the heterogeneity between studies and the change index assigned to heterogeneity I2. Due to the heterogeneity between the studies, the random effects model was used to combine the results of the studies. All statistical analyses were performed by STATA version 14 software using meta-analysis commands. Reports indicated that the prevalence of hallucinations in Parkinson's patients in 32 studies was 28% (0.22-0.34 = 95%CI). The highest prevalence was 34% and 95% CI = 0.07- 0.61 in developing countries and 27% with CI = 0.33-0.21 in developed countries. Reports showed the prevalence in men was 30% (CI = 0.22-0.38) and in women 23% (95% CI = 0.14-0.31). Considering the relatively high prevalence of hallucinations in these patients, checking up for the presence of hallucinations on every visit of Parkinson's patients is recommended, and providing appropriate treatment for that is necessary.


Effect of tryptophan on hepatic nuclear free and engaged RNA-polymerases in young and adult rats.
Whereas in young rats (2 weeks old), administration of typtophan produced marked enhancement in the activity of both engaged and free polymerases of nuclei, in adult rats (10 weeks old) only the engaged polymerases showed higher activities following tryptophan force-feeding.


Long-term efficacy of high-dose diltiazem for chronic stable angina pectoris: 16-month serial studies with placebo controls.
In order to assess the long-term efficacy of diltiazem for the treatment of angina pectoris, eight patients with chronic stable exertional angina who were previously entered into a 4-month randomized, double-blind placebo controlled study, were studied for an additional 12-months. The patients continued to take diltiazem, 360 mg/day, and underwent treadmill exercise testing after 10 and 16 months of therapy. A single-blind placebo week was introduced after 16 months and a treadmill test was performed at the end of this week. Diltiazem therapy continued to augment exercise duration until 0.1 mV of ECG ST depression at 10 and 16 months as compared to the final placebo period: 573 +/- 133 (SD) seconds at 10 months; 565 +/- 148 seconds at 16 months; vs 431 +/- 151 seconds at final placebo (both p less than 0.001). Also, the time to angina pectoris was prolonged on diltiazem by 181 seconds at 16 months (p less than 0.01) and the total duration of exercise was increased by 101 seconds (p less than 0.001) as compared to placebo. In addition, angina frequency decreased from 17 +/- 11 attacks/week on placebo to 0.6 +/- 0.6 attacks/week during diltiazem therapy at 16 months. Two of the eight patients noted mild pedal edema, but no other adverse effects were experienced. Thus diltiazem, 360 mg/day, can be an effective single agent for the long-term treatment of chronic stable angina pectoris.


Impact of the COVID-19 pandemic on health care and daily life of patients with rare diseases from the perspective of patient organizations - a qualitative interview study.
During the COVID-19 pandemic people affected by rare diseases (RD) or caregiver of affected children have faced additional challenges. The pandemic has affected physical and mental health, social life and has led to financial consequences. Our objectives were to identify the impact of COVID-19 (1) on health care and (2) on daily life and participation of patients with RDs or caregivers from the perspective of representatives of patient organizations. Moreover, we explored their perspective on experiences of pandemic stress and resources during the pandemic. We conducted 18 semi-structured interviews with representatives of patient organizations (e.g. chairperson, members of the steering committee), who were asked about the experiences of their members. The interviews were transcribed verbatim and analyzed using the framework approach. We contextualized our findings on the basis of the International Classification of Functioning, Disability and Health (ICF) model and adapted it according to identified subthemes. Patients and caregivers were confronted with aspects of pandemic stress such as lack of information, access and information regarding vaccination and being a risk group for COVID-19 infection. Physical and mental functioning was reported to be negatively impacted. Lock downs and contact restrictions led, e.g., to increasing lack of nursing services or lack of necessary informal support. Participation e.g. in social life and work was reduced. Health care services including medical care and supportive care as well as additional therapies were disrupted and greater effort was necessary to organize care. According to participants, central resources were informal support networks, digitalization, patient organizations and individual characteristics. Our study highlights the consequences of the COVID-19 pandemic on the situation of people affected by RDs and caregivers. Contextualization of the results into the biopsychosocial model reinforces the impact of the pandemic on health care as well as daily life and participation. Major challenges and difficulties were experienced during lockdowns and contact restrictions. Depending on the risk of an infection with COVID-19, certain patient groups were still isolated and reduced social contacts or still followed strict hygienic measures (e.g., wearing medical masks). Future pandemic control measures, e.g. on lockdowns and closing facilities, should consider the challenges of people with RDs and caregivers of affected children.


Systematic measuring cortical thickness in tibiae for bio-mechanical analysis.
Measuring the thickness of cortical bone tissue helps diagnose bone diseases or monitor the progress of different treatments. This type of measurement can be performed visually from CAT images by a radiologist or by semi-automatic algorithms from Hounsfield values. This article proposes a mechanism capable of measuring thickness over the entire bone surface, aligning and orienting all the images in the same direction to have comparable references and reduce human intervention to a minimum. The objective is to batch process large numbers of patients' CAT images obtaining thicknesses profiles of their cortical tissue to be used in many applications. Classical morphological and Deep Learning segmentation is used to extract the area of interest, filtering and interpolation to clean the bones and contour detection and Signed Distance Functions to measure the cortical Thickness. The alignment of the set of bones is achieved by detecting their longitudinal direction, and the orientation is performed by computing their principal component of the center of mass slice. The method processed in an unattended manner 67% of the patients in the first run and 100% in the second run. The difference in the thickness values between the values provided by the algorithm and the measures done by a radiologist was, on average, 0.25 millimetres with a standard deviation of 0.2. Measuring the cortical thickness of a bone would allow us to prepare accurate traumatological surgeries or study their structural properties. Obtaining thickness profiles of an extensive set of patients opens the way for numerous studies to be carried out to find patterns between bone thickness and the patients' medical, social or demographic variables.


Preimplantation genetic screening (PGS) with Comparative genomic hybridization (CGH) following day 3 single cell blastomere biopsy markedly improves IVF outcomes while lowering multiple pregnancies and miscarriages.
To determine benefits of cleavage-stage preimplantation genetic screening (PGS) by array comparative genomic hybridization (CGH). A retrospective case-control study was performed at a tertiary care university-affiliated medical center. Implantation rate was looked at as a primary outcome. Secondary outcomes included clinical and ongoing pregnancy rates, as well as multiple pregnancy and miscarriage rates. Thirty five patients underwent 39 fresh cycles with PGS by aCGH and 311 similar patients underwent 394 invitro fertilization cycles. The implantation rate in the CGH group doubled when compared to the control group (52.63 % vs. 19.15 %, p = < 0.001), clinical pregnancy rate was higher (69.23 % vs. 43.91 %, p = 0.0002), ongoing pregnancy rate almost doubled (61.54 % vs. 32.49 %, p = < 0.0001), multiple pregnancy rate decreased (8.33 % vs. 34.38 %, p = 0.0082) and miscarriage rate trended lower (11.11 % vs. 26.01 %, p = 0.13). Cleavage stage PGS with CGH is a feasible and safe option for aneuploidy screening that shows excellent outcomes when used in fresh cycles. This is the first report of cleavage stage PGS by CGH showing improved ongoing pregnancy rates.


Retraction notice to "Genome-wide analysis of the B-box gene family in the sweet potato wild ancestor Ipomoea trifida and determination of the function of IbBBX28 in the regulation of flowering time of Arabidopsis" [Plant Physiol. Biochem. 188 (2022) 109-122].
This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editors-in-Chief. A large part of the article is highly similar to the paper previously published by Wenqian Hou, Lei Ren, Yang Zhang, Haoyun Sun, Tianye Shi, Yulan Gu, Aimin Wang, Daifu Ma, Zongyun Li and Lei Zhang in Scientia Horticulturae 288 (2021) 110374 https://doi.org/10.1016/j.scienta.2021.110374. In particular, a large part of the two articles shows a study on the same gene family in the same plant, with similar methodological approaches, resulting in a series of highly similar figures. One of the conditions of submission of a paper for publication is that authors declare explicitly that their work is original and has not appeared in a publication elsewhere. Re-use of any data should be appropriately cited. As such this article represents a severe abuse of the scientific publishing system. The scientific community takes a very strong view on this matter and apologies are offered to readers of the journal that this was not detected during the submission process.


Study on the effects of different dimerization positions on biological activity of partial d-Amino acid substitution analogues of Anoplin.
Antimicrobial peptides have recently attracted much attention due to their broad-spectrum antimicrobial activity, rapid microbial effects, and minimal tendency toward resistance development. In this study, a series of new C-C terminals and C-N terminals dimer peptides were designed and synthesized by intermolecular dimerization of the partial d-amino acid substitution analogues of Anoplin, and the effects of different dimerization positions on biological activity were researched. The antimicrobial activity and stability of the new C-C terminals and C-N terminals dimer peptides were significantly improved compared with their parent peptide Anoplin. They displayed no obvious hemolytic activity and lower cytotoxicity, with a higher therapeutic index. Furthermore, the new dimer peptides not only enabled to rapidly disrupt bacterial membrane and damage its integrity which was different from conventional antibiotics but also penetrated bacterial membrane into binding to intracellular genomic DNA. More importantly, the new dimer peptides showed excellent antimicrobial activity against multidrug-resistant strains isolated from clinics in contrast to conventional antibiotics with low tendency to develop the bacterial resistance, besides they exhibited better anti-biofilm activity than antibiotic Rifampicin. Interestingly, the C-N terminals dimer peptides were superior to C-C terminals ones in antimicrobial and anti-biofilm activity, therapeutic index, outer membrane permeability, and DNA binding ability, whereas there were no noteworthy effects in different dimerization positions on stability. Thus, these data suggested that dimerization in different positions represented a potent strategy to develop novel antimicrobial agents for fighting against increasing bacterial resistance.


Staphylokinase reduces plasmin formation by endogenous plasminogen activators.
Hyperfibrinolysis is a consequence of imbalance between fibrinolytic activators and their inhibitors. Increased levels of circulating plasminogen (Plg) activators such as tissue- or urokinase-type plasminogen activators (tPA or uPA respectively) are the most common causes of hyperfibrinolysis, occasionally causing major hemorrhages. We found that staphylokinase (SAK), a well-known Plg activator of bacterial origin, inhibits Plg activation mediated by endogenous tPA and uPA. Furthermore, mixture of SAK with tPA led to a significantly reduced Plg-dependent fibrinolysis. This inhibitory effect was exerted through direct action of SAK on Plg rather than indirectly on tPA or uPA. Inhibition of Plg activation by SAK is readily abrogated by interaction of SAK with human neutrophil peptides (HNPs). Finally, we show that NH2-terminal residues of SAK are important for the inhibitory effect of SAK on tPA- and uPA-mediated Plg activation. In conclusion, SAK reduces tPA/uPA-mediated Plg activation by means of SAK.Plg complex formation, consequently downregulating tPA/uPA-induced fibrinolysis.


Effect of Vitamin C Supplements on Respiratory Tract Infections: A Systematic Review and Meta-Analysis.
Respiratory tract infections are a primary cause of illness and mortality over the world. This study was aimed to investigate the effectiveness of vitamin C supplementation in preventing and treating respiratory tract infections. We used the Cochrane, PubMed, and MEDLINE Ovid databases to conduct our search. The inclusion criteria were placebo-controlled trials. Random effects meta-analyses were performed to measure the pooled effects of vitamin C supplementation on the incidence, severity, and duration of respiratory illness. We found ten studies that met our inclusion criteria out of a total of 2758. The pooled risk ratio (RR) of developing respiratory illness when taking vitamin C regularly across the study period was 0.94 (with a 95% confidence interval of 0.87 to 1.01) which found that supplementing with vitamin C lowers the occurrence of illness. This effect, however, was statistically insignificant (P= 0.09). This study showed that vitamin C supplementation had no consistent effect on the severity of respiratory illness (SMD 0.14, 95% CI -0.02 to 0.30: I2 = 22%, P=0.09). However, our study revealed that vitamin C group had a considerably shorter duration of respiratory infection (SMD -0.36, 95% CI -0.62 to -0.09, P = 0.01). Benefits of normal vitamin C supplementation for reducing the duration of respiratory tract illness were supported by our meta-analysis findings. Since few trials have examined the effects of therapeutic supplementation, further research is needed in this area.


Exposure to lead and cadmium of children living near a lead smelter at Lavrion, Greece.
Exposure of children of the Greek town of Lavrion to lead and cadmium has been evaluated by determination of the levels of lead (PbB) and cadmium (CdB) in the blood of 514 children living in the town. The Lavrion area is heavily polluted by lead owing to the presence of a large lead-zinc mining and smelting industrial complex. The industrial PbB level (geometric mean) was 21.7 micrograms dl-1 (range 7.4-79.8 micrograms dl-1). The CdB levels were, on average, 0.36 micrograms l-1 (range 0.1-3.1 micrograms l-1). The results were evaluated with respect to a number of constitutional, social and environmental variables, such as age, gender, distance of the school from the lead-zinc mining and smelting complex, and occupation and education of the father.


Subtractive hybridization and random arbitrarily primed PCR analyses of a benzoate-assimilating bacterium, Desulfotignum balticum.
Subtractive hybridization (SH) and random arbitrarily primed PCR (RAP-PCR) were used to detect genes involved in anaerobic benzoate degradation by Desulfotignum balticum. Through SH, we obtained 121 DNA sequences specific for D. balticum but not for D. phosphitoxidans (a non-benzoate-assimilating species). Furthermore, RAP-PCR analysis showed that a 651-bp DNA fragment, having 55% homology with the solute-binding protein of the ABC transporter system in Methanosarcina barkeri, was expressed when D. balticum was grown on benzoate, but not on pyruvate. By shotgun sequencing of the fosmid clone (38,071 bp) containing the DNA fragment, 33 open reading frames (ORFs) and two incomplete ORFs were annotated, and several genes within this region corresponded to the DNA fragments obtained by SH. An 11.3-kb gene cluster (ORF10-17) revealed through reverse transcription-PCR showed homology with the ABC transporter system and TonB-dependent receptors, both of which are presumably involved in the uptake of siderophore/heme/vitamin B(12), and was expressed in response to growth on benzoate.


Variation, transcription and circular RNAs of the mitochondrial gene for subunit I of cytochrome c oxidase.
The gene for subunit I of cytochrome c oxidase, contained within the OX13 region of yeast mitochondrial DNA, is split and shows a remarkable variation in structure, which is strain-dependent. The most complex form so far characterized is that of the Saccharomyces cerevisiae strain KL14-4A, in which nine or possibly ten exons are separated by eight to nine introns. At least four of these are facultative, two being absent from S. cerevisiae strain D273-10B (sequenced by Bonitz et al., 1980) and a further two lacking from the gene in Saccharomyces carlsbergensis. The complexity of the gene in KL14-4A is also reflected in its transcript pattern. RNA blot hybridization with isolated and cloned DNA fragments of the OX13 region permits visualization of more than 60 RNAs, which show overlapping and discontinuous hybridization behaviour. In the less complex strains D273-10B and S. carlsbergensis, this number is 20 and 11, respectively. These RNAs are most likely intermediates in processing events leading to the appearance of the mature messenger RNA for cytochrome c oxidase subunit I, which we identify as a 2100-nucleotide transcript (18SE). Most of the processing events are dependent on mitochondrial protein synthesis and do not constitute a single obligatory processing pathway. Like other yeast mitochondrial mRNAs, the 18 S RNA contains a long, untranslated 5' flanking sequence (approximately 400 nucleotides). One unusual aspect of splicing events involving OX13 transcripts is the accumulation of three of the excised introns as single-stranded RNA circles. These abundant and stable transcripts appear to be covalently closed. The simplest assumption is that they arise as (by)-products of splicing, but secondary ligation events have not been excluded. Their function is as yet unknown.


Who Places High Value on the Uterus? A Cross-sectional Survey Study Evaluating Predictors for Uterine Preservation.
To determine predictors for placing high value on the uterus in patients who no longer desire fertility. The secondary objective was to identify reasons for placing high value on the uterus. Cross-sectional survey study. Three hospitals within a large healthcare system in the United States. New patients ≥45 years old seeking care for benign gynecologic conditions, including abnormal uterine bleeding, uterine myomas, pelvic organ prolapse, endometriosis, or pelvic pain. None. The primary outcome was the summative score of the validated Value of Uterus (VALUS) instrument for measuring value placed on the uterus and the validated visual analog scale with the question "how important is it to you to keep your uterus when you have a gynecologic condition?" A total of 163 surveys were returned for analysis (79.2%). Using the VALUS cutoff, 64 patients (45.7%) were considered to have low value for their uterus (VALUS score <14), whereas 76 patients (54.3%) were considered to have high value for their uterus (VALUS score ≥14). The adjusted odds of placing high value for the uterus was 5.06 times higher among those who wanted to be sexually active in the future than those who do not desire to be sexually active (95% confidence interval, 1.55-16.52, p = .01). Patients who are sexually active have 3.94 higher adjusted odds of placing high value on the uterus than those who are not sexually active and do not desire to be (95% confidence interval, 1.36-11.43; p = .01). Race, religion, and personal history of cancer were not statistically significant. Patients who highly value the uterus were highly motivated by the desire to be sexually active. Nonwhite race, religion, and personal history of cancer were not predictors for placing high value on uterine preservation.


Distribution and sub-cellular localization of drug metabolizing enzymes in the skin.
The distribution and sub-cellular localization of cytochrome P-450-dependent mono-oxygenase activity in the skin of adult hairless mice has been investigated using ethoxycoumarin, ethoxyresorufin and benzpyrene as substrates. Highest levels of mono-oxygenase activity per unit area of skin were found in the dermis, both before and after induction with benzanthracene. Subcellularly, mono-oxygenase activity was predominantly microsomal and, prior to induction, located entirely in the smooth microsomal fraction.


Control of vascular tube morphogenesis and maturation in 3D extracellular matrices by endothelial cells and pericytes.
An important advance using in vitro EC tube morphogenesis and maturation models has been the development of systems using serum-free defined media. Using this approach, the growth factors and cytokines which are actually necessary for these events can be determined. The first model developed by our laboratory was such a system where we showed that phorbol ester was needed in order to promote survival and tube morphogenesis in 3D collagen matrices. Recently, we have developed a new system in which the hematopoietic stem cell cytokines, stem cell factor (SCF), interleukin-3 (IL-3), and stromal derived factor-1α (SDF-1α) were added in conjunction with FGF-2 to promote human EC tube morphogenesis in 3D collagen matrices under serum-free defined conditions. This new model using SCF, IL-3, SDF-1α, and FGF-2 also works well following the addition of pericytes where EC tube formation occurs, pericytes are recruited to the tubes, and vascular basement membrane matrix assembly occurs following EC-pericyte interactions. In this chapter, we describe several in vitro assay models that we routinely utilize to investigate the molecular requirements that are critical to EC tube formation and maturation events in 3D extracellular matrix environments.


Cyclodextrin-based strategies for removal of persistent organic pollutants.
Water could be considered one of the essential natural resources on our planet. However, water contamination has become virtually ubiquitous during the past decades due to several factors, e.g., world population growth, water use in chemical production, and agriculture. Among contaminants, persistent organic pollutants (POPs) were widely spread during the last few decades and are now present all around the world. POPs are connected with several different illnesses, and therefore the removal of these pollutants from water is a challenge for coming years. Cyclodextrins (CDs) are environmentally friendly and cheap pollutant adsorbents due to their peculiar physicochemical properties. Moreover, CDs are relatively easy to use, and many techniques are reported for their functionalization. In addition, several CD derivatives are known and some of them are commercially available. CDs and CD-functionalized materials could be used for removal of different pollutants by using various methods, e.g., adsorption/extraction, soil washing, and electrokinetic/catalytic processes. This is the first review article that provides an overview of CDs/CD-based materials and their use in the remediation of POPs listed under the Stockholm Convention. The mechanisms of the POPs removal by CDs are also discussed.


Excitatory amino acid transporter 1 supports adult hippocampal neural stem cell self-renewal.
Within the adult mammalian dentate gyrus (DG) of the hippocampus, glutamate stimulates neural stem cell (NSC) self-renewing proliferation, providing a link between adult neurogenesis and local circuit activity. Here, we show that glutamate-induced self-renewal of adult DG NSCs requires glutamate transport via excitatory amino acid transporter 1 (EAAT1) to stimulate lipogenesis. Loss of EAAT1 prevented glutamate-induced self-renewing proliferation of NSCs in vitro and in vivo, with little role evident for canonical glutamate receptors. Transcriptomics and further pathway manipulation revealed that glutamate simulation of NSCs relied on EAAT1 transport-stimulated lipogenesis. Our findings demonstrate a critical, direct role for EAAT1 in stimulating NSCs to support neurogenesis in adulthood, thereby providing insights into a non-canonical mechanism by which NSCs sense and respond to their niche.


Phosphorylation activity associated with plasma membrane-cortical granule complex of Xenopus laevis oocytes.
Plasma membrane-cortical granule complexes from Xenopus laevis oocytes were isolated by a differential centrifugation procedure. Membrane preparations incorporated 32P from [gamma-32P]ATP predominantly into three protein bands of Mr 128,000, 54,000 and 29,000, as determined by sodium dodecyl-polyacrylamide gel electrophoresis. Progesterone and insulin, added in vitro to membranes, did not produce any change in this phosphorylation pattern, contrary to some previously published results. The availability of enriched membrane complexes should permit further biochemical dissection of signal transduction pathways that are activated during oocyte maturation and fertilization.


High-resolution mapping of the rainfall runoff pollution: case study of Shiwuli River watershed, China.
Rainfall runoff is the key factor of water quality deterioration in highly urbanized area, which is characterized by intensive human activities and frequent extreme weather events. Urban landscape system is composed of highly diverse and heterogeneous land patches, which makes the effective management of urban runoff pollution difficult. Therefore, high-resolution land-use data is imperative for the identification and analysis of spatial-temporal characteristics of urban runoff pollution. In this study, Shiwuli River watershed, a rapidly urbanizing area in China, is selected as the study area. We first interpret nine kinds of land-use types with a high-resolution remote sensing data of 2 m [Formula: see text] 2 m. Then, a localized Soil Conservation Service model based on field observation and rainfall experiments is applied to map the spatial-temporal pattern of runoff pollution. The results indicate that the COD, NH3-N, TP, and TN load generated by the runoff in the watershed accounted for 23.4%, 3.7%, 8.2%, and 9.0% of the total pollution load in 2016, respectively. Furthermore, the spatial-temporal pattern of the assessed runoff pollution was mainly subject to the distribution of rainfall and land-use patterns. We suggest that the sponge city construction combined with surface pollution control is an effective way to reduce the runoff pollution. This study highlights the necessity to identify spatial-temporal hotspots in developing precise pollution control measures, which provides valuable information for pollution control policy-making in Shiwuli River watershed and could serve as a reference for other river watersheds undergoing rapid urbanization.


Toxoplasma gondii: serological characterization and immunogenicity of recombinant surface antigen 2 (SAG2) expressed in the yeast Pichia pastoris.
The full length surface antigen 2 (SAG2) gene of the protozoan parasite Toxoplasma gondii was cloned and intracellularly expressed in the Pichia pastoris expression system. The molecular weight of the expressed recombinant SAG2 (36 kDa) was much larger than the native SAG2 (22 kDa). This discrepancy in size was due to hyperglycosylation, as deglycosylation assay reduced the size of the recombinant SAG2 to 22 kDa. Despite being hyperglycosylated, the recombinant SAG2 reacted strongly with pooled anti-Toxoplasma human serum, pooled anti-Toxoplasma mouse serum and a SAG2-specific monoclonal antibody. The glycosylated recombinant SAG2 was further evaluated in Western blot and in-house enzyme-linked immunosorbent assay (ELISA) using 80 human serum samples, including confirmed early acute (IgM positive, IgG negative; n=20), acute (IgM positive, IgG positive; n=20) and chronic (IgM negative, IgG positive; n=20) toxoplasmosis patients, and toxoplasmosis negative control patients (n=20). Results of the Western blot showed that the recombinant SAG2 reacted with all 60 samples of the toxoplasmosis cases but not with the Toxoplasma-negative samples. The sensitivity of in-house ELISA was 80%, 95% and 100% for early acute, acute and chronic patients' serum samples, respectively. Vaccination study showed that serum from mice immunised with the glycosylated recombinant SAG2 reacted specifically with the native SAG2 of T. gondii. The mice were significantly protected against lethal challenge with live T. gondii RH strain tachyzoites (P<0.01) and their survival time was increased compared to controls. Therefore, the present study shows that the P. pastoris-derived recombinant SAG2 was specific and suitable for use as antigen for detecting anti-Toxoplasma IgG and IgM antibodies. The vaccination study showed that recombinant SAG2 protein was immunoprotective in mice against lethal challenge.


Fall risk management using clinical decision support in a rural nursing-led primary care practice.
The leading cause of injuries among older adults in the United States is unintentional falls. The American Geriatrics Society/British Geriatrics Society promote fall risk management in primary care; however, this is challenging in low-resource settings. Archer Family Health Care (AFHC), an Advanced Practice Registered Nurse (APRN)-managed and federally designated rural health clinic, identified a care gap with falls adherence to guidelines for patients at higher risk for falls. The aim of this quality improvement effort was to integrate an evidence-based fall risk management tool in a rural nurse-managed primary care practice. A standardized fall risk management process with a new brief paper-based clinical decision support (CDS) tool was developed and tested in two phases. Phase 1 focused on developing a fall risk management CDS tool, identifying the primary care visit workflow, communicating the workflow patterns to the AFHC staff, and collaborating with the staff to identify when and who should implement the tool. Phase 2 focused on implementation of the fall risk management CDS tool into standard practice among older adults aged 65 years and older. We found that integrating the tool did not disrupt the workflow of primary care visits at AFHC. The most common recommended intervention for patients at risk of falling was daily vitamin D supplementation. This project revealed that it is feasible to introduce a brief fall risk management decision support tool in an APRN-managed rural primary care practice.


Effect of submaximal exercise at low temperatures on pulmonary function in healthy young men.
In order to understand more fully the effect on pulmonary function of whole body exposure to cold during submaximal exercise, we measured pulmonary function indices in ten healthy male students and ten healthy male forestry workers of similar age following submaximal treadmill walking at different temperatures in a climatic chamber. After measuring the maximal aerobic capacity with a cycle ergometer test, the subjects had to walk on four separate occasions in the climatic chamber at an intensity of 70%-75% of their individual maximal heart rate; the first at normal room temperature and then randomly, either at 0 degrees C or at -20 degrees C, and vice versa. The duration of each walk was 8 min. Finally, each subject had to walk in the chamber at -20 degrees C for 17 min. Flow volume spirometry was performed at room temperature 1, 5, 10, and 20 min after exercise and the values were compared to baseline values taken prior to the last walking test. There were only minor changes in pulmonary function indices following exercise at different temperatures. Only one student showed a reduction of over 15% in peak expiratory flow rate after an 8-min walk at -20 degrees C. It seems that submaximal exercise of short duration, even at a temperature as low as -20 degrees C, does not impair pulmonary function in healthy young men.


Imagination nourishes dietetic practice. 2005 Ryley-Jeffs Memorial Lecture.
Albert Einstein once stated that imagination is more important than knowledge. How important is imagination to the dietetic profession? What have been the imaginations of dietitians over the years? Where would we be today without these imaginations? Can imagination be fostered and developed? What future imaginations will shape the dietetic profession? This article explores the phenomenon of imagination and why it is important to dietetic practice.


Nutrition support during extracorporeal membrane oxygenation (ECMO) in adults: a retrospective audit of 86 patients.
Extracorporeal membrane oxygenation (ECMO) is increasingly being used to support critically ill patients with severe cardiac and/or respiratory failure. It has been claimed that the resulting haemodynamic alterations, particularly in venoarterial ECMO, mean that enteral feeding is unsafe and/or poorly tolerated. This study aims to investigate this question and to identify any barriers to optimal nutrition. Data were retrospectively collected for 86 patients who received ECMO between January 2007 and July 2012 in a tertiary critical care unit/ECMO referral centre. All were fed using existing protocols that emphasise early enteral feeding in preference over parenteral or delayed enteral nutrition. Thirty-one patients required ECMO for cardiac failure, and all of these received venoarterial ECMO; the remainder received venovenous ECMO. Enteral feeds started for all patients at average 13.1 h [standard deviation (SD) 16.7 h] after ICU admission, reaching goal rate on day 2.6 (SD 1.4). Thirty-three patients experienced significant feeding intolerance during the first 5 days, but of these 20 were managed effectively with prokinetic medications; 18 required parenteral nutrition to supplement inadequately tolerated tube feeds. Intolerance did not differ between ECMO modes. Overall patients tolerated 79.7% of goal nutrition each day in the first 2 weeks. Enteral feeding can be well tolerated by patients who are receiving ECMO, whether in venovenous or venoarterial mode. ECMO should not exclude patients from receiving the well-documented benefits of early enteral feeding in critical illness.


Effect of preoperative chemotherapy on distal spread of low rectal cancer located close to the anus.
This study aimed to clarify the frequency of distal spread and the optimal distal margin after preoperative chemotherapy for advanced low rectal cancer. The study included patients with advanced lower rectal cancer who received preoperative chemotherapy and underwent surgery during 2012-2015. We investigated the distal spread of tumor cells, defined as the distal distance from the intramucosal distal tumor edge to the farthest tumor cells located under the submucosal layer. Clinical characteristics were compared for distal spreads ≥ 10 and < 10 mm, and risk factors for distal spread ≥ 10 mm were investigated. Of the 71 patients, 42 (59%) showed distal spread. Distal spreads of 1-9, 10-19, and ≥ 20 mm were observed in 27 (38%), 11 (15%), and 4 (6%) patients, respectively. Multivariate analysis revealed two independent risk factors for distal spread ≥ 10 mm after preoperative chemotherapy. The first risk factor is the presence of different therapeutic effects between the mucosal and deeper layers (meaning that superficial tumor shrinkage was evident on colonoscopy, but little tumor shrinkage was evident on magnetic resonance imaging) (odds ratio, 11.6; 95% CI, 2.22-61.3). The second risk factor is poorly differentiated or mucinous adenocarcinoma (odds ratio, 8.86; 95% CI, 1.58-49.9). A distal margin of 20 mm is required (10 mm is insufficient) for advanced lower rectal cancer patients who receive preoperative chemotherapy followed by surgery. Independent risk factors for distal spread ≥ 10 mm include (1) the presence of different therapeutic effects between mucosal and deeper layers and (2) poorly differentiated or mucinous adenocarcinomas.


Galanin receptor binding sites in the temporal and occipital cortex are minimally affected in Alzheimer's disease.
Galanin receptor binding sites were examined in the inferior temporal and medial occipital gyri of patients with Alzheimer's disease (AD) and matched control subjects using quantitative autoradiography. In the inferior temporal gyrus, galanin binding was reduced selectively in layers V-VI of the AD cases compared to controls, the magnitude of the reduction (45%) being similar to that of choline acetyltransferase activity (40%) in this region. In the medial occipital gyrus, galanin binding in the AD cases was not different from controls in any cortical layer despite a reduction in choline acetyltransferase activity. Galanin binding did not correlate with the densities of neuritic plaques in either temporal or occipital gyri. Thus, despite a significant cortical cholinergic deficit in AD, there is an anatomically selective reduction of cortical galanin receptor binding sites suggesting that the majority of galanin receptors are not located on cholinergic terminals in the human cerebral cortex.


An approach towards experimental cDNA sequence determination of predicted genes: an example from Arabidopsis U3-55k homologues.
Experimental cDNA sequence determinations lag behind in silico gene structure predictions in some recently sequenced genomes. This may be due in part to low transcript abundance and/or the severely spatio-temporarily restricted expression pattern of some genes. Here we characterize the predicted repressed gene of Arabidopsis thaliana (At4g21130) that encodes a homologue of the Arabidopsis U3-55K-like protein (At4g05410) and of the U3-55K (RNU3IP2, Rrp9p) proteins from other eukaryotes. In man and yeast, U3-55K is involved in the processing of the pre-ribosomal RNA. Here we show that treatment with inhibitors of histone deacetylases (trichostatin A, sodium butyrate) or DNA methyltransferases (5-aza-2'-deoxycytidine) induces a low but distinct level of mRNA from the repressed Arabidopsis At4g21130 locus, which can be detected by RT-PCR amplification. Direct sequencing of PCR products reveals the open reading frame that differs, in part, from the hypothetical one and encodes a seven-WD-repeat protein highly conserved when compared to U3-55K proteins from various eukaryotic species. This suggests the conservation of its function. The described approach may help to determine the nucleotide sequences of transcripts from predicted genes with a low level of expression.


[Optimization of methods and improvement of technical means in combination with the nonspecific reparative regeneration stimulator ximedone in dermatoplasty surgery].
Treatment of extensive tissue defects should be complex, whose basis is an early free skin plastic repair with obligatory addition of agents that enhance the reparative capacities of tissues. For this, the authors propose a number of original appliances (forceps, variants of dermoperforators, bandage, etc.) and drugs, such as ximedon (tablets, 5-10% aqueous solution, lanolin-vaseline-based ointment, talc-based powder), whose clinical trials in 300 patients have shown their high therapeutical effect. The time of preparation of the wound surface for dermatoplastic interventions became shorter and the frequency of poor outcomes of operations reduced.


Serum Vitamin D Depends Less on Latitude Than on Skin Color and Dietary Intake During Early Winter in Northern Europe.
To evaluate if dietary vitamin D intake is adequate for sufficient vitamin D status during early winter in children living in Sweden, irrespective of latitude or skin color. As part of a prospective, comparative, 2-center intervention study in northern (63°N) and southern (55°N) Sweden, dietary intake, serum 25-hydroxyvitamin D (S-25(OH) D), associated laboratory variables, and sociodemographic data were studied in 5 to 7-year-old children with fair and dark skin in November and December. Two hundred six children with fair/dark skin were included, 44/41 and 64/57 children in northern and southern Sweden, respectively. Dietary vitamin D intake was higher in northern than southern Sweden (P = 0.001), irrespective of skin color, partly due to higher consumption of fortified foods, but only met 50-70% of national recommendations (10 μg/day). S-25(OH) D was higher in northern than southern Sweden, in children with fair (67 vs 59 nmol/L; P < 0.05) and dark skin (56 vs 42 nmol/L; P < 0.001). S-25(OH) D was lower in dark- than fair-skinned children at both sites (P < 0.01), and below 50 nmol/L in 40 and 75% of dark-skinned children in northern and southern Sweden, respectively. Insufficient vitamin D status was common during early winter in children living in Sweden, particularly in those with dark skin. Although, higher dietary vitamin D intake in northern than southern Sweden attenuated the effects of latitude, a northern country of living combined with darker skin and vitamin D intake below recommendations are important risk factors for vitamin D insufficiency.


Characterization of the Abelson murine leukemia virus-encoded tyrosine-specific protein kinase.
There is sufficient evidence that the A-MuL V protein is a tyrosine-specific protein kinase. There are methods for detecting this kinase activity and this kinase has been expressed in E. coli. Because the information coding for the tyrosine-specific protein kinase is present in normal mouse cells, such an enzyme must have a normal physiologic function. The elucidation of this physiologic function and the understanding of the role of this enzyme activity in neoplastic transformation is the challenge of the future.


Genotyping-by-Sequencing Enabled Mapping and Marker Development for the By-2 Potyvirus Resistance Allele in Common Bean.
Since its emergence in 2001, an aphid-transmitted virus disease complex has caused substantial economic losses to snap bean (Phaseolus vulgaris L.) production and processing in the Great Lakes Region of the United States. The general ineffectiveness of chemical control measures for nonpersistently transmitted viruses established an urgent need for the development and deployment of cultivars with resistance to the component viruses. Our objectives were to further characterize the inheritance of resistance to Bean yellow mosaic virus (BYMV), which is conditioned by the By-2 allele, to adapt genotyping-by-sequencing (GBS) to common bean to discover and genotype genome-wide single nucleotide polymorphisms (SNPs) in a set of recombinant inbred lines (RILs) derived from an introgression program, and to enable and validate marker-assisted selection for By-2. We optimized ApeKI for GBS in common bean and retained 7530 high-quality SNPs that segregated in our introgression RILs. A case-control genome-wide association study (GWAS) was used to discover 44 GBS SNPs that were strongly associated with the resistance phenotype and which delimited a 974 kb physical interval on the distal portion of chromosome 2. Seven of these SNPs were converted to single-marker Kompetitive Allele-Specific Polymerase chain reaction (KASP) assays and were demonstrated to be tightly linked to BYMV resistance in an F2 population of 185 individuals. This research enables marker-assisted selection of By-2, provides enhanced resolution for fine mapping, and demonstrates the potential of GBS as a highly efficient, high-throughput genotyping platform for common bean breeding and genetics.


Magnetic and Electronic Structural Properties of the S3 State of Nature's Water Oxidizing Complex: A Combined Study in ELDOR-Detected Nuclear Magnetic Resonance Spectral Simulation and Broken-Symmetry Density Functional Theory.
ELDOR-detected nuclear magnetic resonance (EDNMR) spectral simulations combined with broken-symmetry density functional theory (BS-DFT) calculations are used to obtain and to assign the 55Mn hyperfine coupling constants (hfcs) for modified forms of the water oxidizing complex in the penultimate S3 state of the water oxidation cycle. The study shows that an open cubane form of the core Mn4CaO6 cluster explains the magnetic properties of the dominant S = 3 species in all cases studied experimentally with no need to invoke a closed cubane intermediate possessing a distorted pentacoordinate Mn4 ion as recently suggested. EDNMR simulations found that both the experimental bandwidth and multinuclear transitions may alter relative EDNMR peak intensities, potentially leading to incorrect assignment of hfcs. The implications of these findings for the water oxidation mechanism are discussed.


[The prevalence of the initial forms of cerebrovascular and cardiovascular diseases and their risk factors among the rural population].
An epidemiological analysis was carried out in 704 rural inhabitants. A significant incidence of cardiovascular diseases and their risk factors was found. The incidence of cerebrovascular pathology prevailed over the cardiac. With age hyperlipidemia increased, particularly in males. Therapeutic and prophylactic measures are discussed.


Steven-Johnson Syndrome Induced by Lamotrigine and Valproic Acid in a Pediatric Patient: A Case Report.
Steven-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are part of a spectrum of severe cutaneous adverse reactions, secondary to infections or drug-induced. Although the use of antiseizure medications (ASMs) is a risk factor for the development of SJS/TEN, primary care physicians are not familiar with these cases in some countries. We report a case of SJS associated with ASMs in a nine-year-old girl with a history of difficult-to-control epilepsy, who required adjustment and change in medications.


Methodological considerations and practical recommendations for the application of peripheral arterial tonometry in children and adolescents.
Endothelial dysfunction is recognized as the primum movens in the development of atherosclerosis. Its crucial role in both cardiovascular morbidity and mortality has been confirmed. In the past, research was hampered by the invasive character of endothelial function assessment. The development of non-invasive and feasible techniques to measure endothelial function has facilitated and promoted research in various adult and paediatric subpopulations. To avoid user dependence of flow-mediated dilation (FMD), which evaluates nitric oxide dependent vasodilation in large vessels, a semi-automated, method to assess peripheral microvascular function, called peripheral arterial tonometry (Endo-PAT(®)), was recently introduced. The number of studies using this technique in children and adolescents is rapidly increasing, yet there is no consensus with regard to either measuring protocol or data analysis of peripheral arterial tonometry in children and adolescents. Most paediatric studies simply applied measuring and analysing methodology established in adults, a simplification that may not be appropriate. This paper provides a detailed description of endothelial function assessment using the Endo-PAT for researchers and clinicians. We discuss clinical and methodological considerations and point out the differences between children, adolescents and adults. Finally, the main aim of this paper is to provide recommendations for a standardised application of Endo-PAT in children and adolescents, as well as for population-specific data analysis methodology.


[Liposomal encapsulation of free anthraquinones in rhizoma et radix rhei and its quality control].
To encapsulate the free anthraquiones in Rhizoma et Radix Rhei (rhein, chrysophanol, physcione, emodin and aloeemodin) in liposomes and characterize the liposomes. The liposomes were separated from free drug with sephadex G-50 with HPLC determination of the free anthraquinones in liposomes, then its entrapment efficiency was calculated. Ethanol injection method was used to prepare the liposomes with the addition of a suitable amount Ca2+ in the medium. The influence of Ca2+ on the entrapment efficiency of free anthraquinones in Rhizoma et Radix Rhei in liposomes has been studied. Dynamic laser scatterometer and transmission electron microscopy were used to study the sizes and morphology of the liposomes. The entrapment efficiency of rhein, chrysophanol, physcione, emodin and aloeemodin in liposomes were 27.63%, 85.16%,100.14%, 99.77% and 66.16%, respectively. The total entrapment efficiency of free anthraquinones in Rhizoma et Radix Rhei was 80.67%. Ca2+ could greatly promote the liposomal encapsulation of free anthraquinones in Rhizoma et Radix Rhei from 13.98% - 23.78% to 81.42% - 89.43%. The liposomes were white spheres and its average size was mainly concentrated on 340 nm. The entrapment efficiency of the liposomal encapsulation of free anthraquinones in Rhizoma et Radix Rhei is high. The quality control method is simple and rapid with good repeatability.


[Combined therapy with quinapril, an ACE inhibitor, and valsartan, a type 1 angiotensin II receptors blocker, for moderate chronic cardiac failure may raise the degree of neurohormonal block and improve 24-h heart rate variability compared to the effect of monotherapy (data from the trial SADKO-CHF)].
To compare effects of various regimens of long-term monotherapy with quinapril (an ACE inhibitor) and valsartan (a type 1 angiotensin II receptors blocker) and combined therapy with these drugs on the activity of heurohormonal systems and 24-h heart rate variability (HRV) in patients with stable moderate chronic cardiac failure (CCF). A total of 80 patients with FC II-III CCF secondary to coronary heart disease (CHD), delated cardiomyopathy (DCMP) and decompensated hypertensive heart (49%/47%/4%) were randomized into 3 groups. Group 1 patients (n = 28) received quinapril, group 2 (n = 26)--valsartan, group 3 (n = 26)--quinapril + valsartan. The levels of norepinephrine (NE), angiotensin II (AT II), activity of plasmic renin (PR), aldosteron (AS), plasmic cerebral sodiumuretic peptide (CSUP) were measured and ECG with determination of HRV and heart rate disturbances was made before and 3, 6 months after the treatment. NE concentration lowered most significantly in group 1 (from 630 to 405 pg/ml) vs groups 2 and 3 (from 525 to 490 pg/ml and from 525 to 480 pg/ml, respectively). A 6-month treatment induced significant changes neither in concentrations of AT II nor AC. ATII concentrations rose 2-fold in group III (from 11.9 to 24.3 pg/ml) under a parallel rise of PR activity from 0.7 to 2.5 ng/ml/h and a fall in AS level from 132 to 83 pg/ml. In group 2 AS diminished also (from 165 to 126 pg/ml). CSUP decreased in all the groups, but significantly only in groups II and III (from 350 to 237 and 322 to 204 pg/ml after 6 months of treatment, respectively). Significant changes of 24-h HRV (both spectral and temporary) were observed in group 1 after 3 months of treatment. These changes lost significance to the end of the treatment. In groups 2 and 3 the changes were less pronounced. Quinapril is more potent than valsartan and quinapril + valsartan combination in relation to activity of sympathico-adrenal system and HRV in patients with moderate stable CCF. Long-term therapy with valsartan does not improve parameters of HRV in moderate CCF. If CCF patients take quinapril for a long time, they develop the effect of disappearing block of AS synthesis and reactivation of A TII production. The best mechanism of long-term control over the activity of renin-angiotensin-aldosteron system in patients with moderate CCF is combination of quinapril with valsartan.


How to succeed in dermatologic surgery.
This article explores the many facets of success within dermatologic surgery. We incorporate advice from both dermatologists and dermatologic surgeons to provide overall advice and strategies for success. In addition, we discuss specific tips for matching and career development.


MicroRNAs visit the ER.
Little is known about where microRNAs (miRNAs) regulate their targets within the cell. In this issue, Li et al. identify a new player in the plant miRNA pathway that implicates the endoplasmic reticulum in miRNA-mediated gene silencing.


A ligand-free solid-supported system for Sonogashira couplings: applications in nucleoside chemistry.
A mild heterogeneous, ligand-free protocol for Sonogashira and Heck couplings has been developed and used to access several biologically important deoxynucleoside derivatives in a facile manner.


Characterization of a synthetic peroxodiiron(III) protein model complex by nuclear resonance vibrational spectroscopy.
The vibrational spectrum of an η(1),η(1)-1,2-peroxodiiron(III) complex was measured by nuclear resonance vibrational spectroscopy and fit using an empirical force field analysis. Isotopic (18)O(2) labelling studies revealed a feature involving motion of the {Fe(2)(O(2))}(4+) core that was not previously observed by resonance Raman spectroscopy.


Effects of L-carnosine on splenic sympathetic nerve activity and tumor proliferation.
l-Carnosine (β-alanyl-l-histidine), a dipeptide of the amino acids β-alanine and histidine, is found in mammalian tissues including those in the central nervous system and in skeletal muscles. In the present study, we examined the effects of intraduodenal (ID) injection of l-carnosine on splenic sympathetic nerve activity (splenic-SNA) in urethane-anesthetized rats and found that ID injection of 3.3mg/kg of body weight of l-carnosine significantly suppressed splenic-SNA. Since it has been suggested that splenic-SNA reduction increases natural killer (NK) activity of splenic cells, which in turn elevates tumor immunity, we then investigated the effect of l-carnosine on the proliferation of human colon cancer cells transplanted into athymic nude mice. The findings of this study revealed that 1mg/mL of l-carnosine solution given as the only drinking water inhibited tumor proliferation. These results suggest that l-carnosine suppresses splenic-SNA and inhibits cancer cell proliferation, probably by elevating NK activity.


Cumulative reduction of primary skin tumor growth in UV-irradiated mice by the combination of retinyl palmitate and canthaxanthin.
The effects of dietary supplementation with retinyl palmitate, canthaxanthin, or the combination of both, on photocarcinogenesis was determined in pigmented C3H/HeN mice. The basal diet was the American Institute of Nutrition Diet 76A, to which was added 120 IU of retinyl palmitate per g diet, 1% canthaxanthin, or the combination of both. Administration of the diets began 18 weeks before the first UVB radiation (280-320 nm) treatment and continued throughout the study. The UV source was a bank of 6 Westinghouse FS40 lamps which delivered to the mice a total dose of 9.9 x 10(5) J/m2, delivered over 24 weeks. These diets significantly reduced the tumor burden per mouse induced by UV irradiation, however they did not influence tumor incidence. The combination of retinyl palmitate plus canthaxanthin was more effective than either agent alone at reducing autochthonous tumor growth, a result which has not been previously reported.


Reversal of the antiviral activity of ribavirin against Sindbis virus in Ae. albopictus mosquito cells.
Earlier work in our laboratory has shown that the replication of Sindbis virus in Aedes albopictus mosquito cells is inhibited by ribavirin (Rbv) and mycophenolic acid (MPA) (Sarver and Stollar (1978) Virology 91, 267-282; Malinoski and Stollar (1980) Virology 102, 473-476). We report here that the antiviral effect of Rbv and MPA can be reversed by depriving infected cells of methionine or isoleucine, or by treating them with fluorodeoxyuridine (FUdR) or cycloleucine. We suggest that, as was the case when the antiviral activity of Rbv was reversed by actinomycin D (Malinoski and Stollar (1981a) Virology 110, 281-291), these effects may be mediated by changes in the GTP pools of treated cells.


Silver(I) nitrate complexes of three tetra-kis-thio-ether-substituted pyrazine ligands: metal-organic chain, network and framework structures.
The reaction of the ligand 2,3,5,6-tetra-kis-[(methyl-sulfanyl)-meth-yl]pyrazine (L1) with silver(I) nitrate led to {[Ag(C12H20N2S4)](NO3)} n , (I), catena-poly[[silver(I)-μ-2,3,5,6-tetra-kis-[(methyl-sulfan-yl)meth-yl]pyrazine] nitrate], a compound with a metal-organic chain structure. The asymmetric unit is composed of two half ligands, located about inversion centres, with one ligand coordinating to the silver atoms in a bis-tridentate manner and the other in a bis-bidentate manner. The charge on the metal atom is compensated for by a free nitrate anion. Hence, the silver atom has a fivefold S3N2 coordination sphere. The reaction of the ligand 2,3,5,6-tetra-kis-[(phenyl-sulfanyl)-meth-yl]pyrazine (L2) with silver(I) nitrate, led to [Ag2(NO3)2(C32H28N2S4)] n , (II), poly[di-μ-nitrato-bis-{μ-2,3,5,6-tetra-kis-[(phenyl-sulfan-yl)meth-yl]pyrazine}disilver], a compound with a metal-organic network structure. The asymmetric unit is composed of half a ligand, located about an inversion centre, that coordinates to the silver atoms in a bis-tridentate manner. The nitrate anion coordinates to the silver atom in a bidentate/monodentate manner, bridging the silver atoms, which therefore have a sixfold S2NO3 coordination sphere. The reaction of the ligand 2,3,5,6-tetra-kis-[(pyridin-2-yl-sulfanyl)-meth-yl]pyrazine (L3) with silver(I) nitrate led to [Ag3(NO3)3(C28H24N6S4)] n , (III), poly[trinitrato{μ6-2,3,5,6-tetra-kis[(pyri-din-2-ylsulfan-yl)meth-yl]pyrazine}-trisilver(I)], a compound with a metal-organic framework structure. The asymmetric unit is composed of half a ligand, located about an inversion centre, that coordinates to the silver atoms in a bis-tridentate manner. One pyridine N atom bridges the monomeric units, so forming a chain structure. Two nitrate O atoms also coordinate to this silver atom, hence it has a sixfold S2N2O2 coordination sphere. The chains are linked via a second silver atom, located on a twofold rotation axis, coordinated by the second pyridine N atom. A second nitrate anion, also lying about the twofold rotation axis, coordinates to this silver atom via an Ag-O bond, hence this second silver atom has a threefold N2O coordination sphere. In the crystal of (I), the nitrate anion plays an essential role in forming C-H⋯O hydrogen bonds that link the metal-organic chains to form a three-dimensional supra-molecular structure. In the crystal of (II), the metal-organic networks (lying parallel to the bc plane) stack up the a-axis direction but there are no significant inter-molecular inter-actions present between the layers. In the crystal of (III), there are a number of C-H⋯O hydrogen bonds present within the metal-organic framework. The role of the nitrate anion in the formation of the coordination polymers is also examined.


Biologically informed variational autoencoders allow predictive modeling of genetic and drug-induced perturbations.
Variational autoencoders (VAEs) have rapidly increased in popularity in biological applications and have already successfully been used on many omic datasets. Their latent space provides a low-dimensional representation of input data, and VAEs have been applied, e.g. for clustering of single-cell transcriptomic data. However, due to their non-linear nature, the patterns that VAEs learn in the latent space remain obscure. Hence, the lower-dimensional data embedding cannot directly be related to input features. To shed light on the inner workings of VAE and enable direct interpretability of the model through its structure, we designed a novel VAE, OntoVAE (Ontology guided VAE) that can incorporate any ontology in its latent space and decoder part and, thus, provide pathway or phenotype activities for the ontology terms. In this work, we demonstrate that OntoVAE can be applied in the context of predictive modeling and show its ability to predict the effects of genetic or drug-induced perturbations using different ontologies and both, bulk and single-cell transcriptomic datasets. Finally, we provide a flexible framework, which can be easily adapted to any ontology and dataset. OntoVAE is available as a python package under https://github.com/hdsu-bioquant/onto-vae.


Human seminal plasma beta-microseminoprotein: its purification, characterization, and immunohistochemical localization.
beta-Microseminoprotein was very efficiently purified from human seminal plasma with only three steps including DEAE-Sephacel and Zinc-chelate Sepharose CL-6B column chromatography. The purified protein was a non-glycoprotein with a molecular weight (M(r)) of 19,000 and 17,000 on gel filtration and reduced SDS-PAGE, respectively. The protein gave six bands from M(r) 15,600 to 25,500 on non-reduced SDS-PAGE. The characterization including the molecular weight, amino acid sequence of N-terminus and concentrations in various body fluids is discussed. Furthermore, the immunohistochemical localization of the protein among various human tissues is demonstrated.


One-pass resurfacing with a combined-mode erbium: YAG/CO2 laser system: a study in 102 patients.
The CO2 and erbium: YAG (Er: YAG) lasers have been used for skin resurfacing. A recently developed system combines pulsed ablative Er: YAG and continuous wave subablative CO2 wavelengths in one console. To assess the potential benefits of this system. The study follows 102 women, skin types I-V, with 26 full face, 48 perioral and 28 periocular resurfacing procedures. The ablative Er: YAG pulse (350 micros, 29 J cm(-2)) is followed immediately by a non-ablative CO2 laser shot (4-6 W, 50 ms) through the same collimated handpiece (3-mm diameter spot), 50% overlapping, repetition rate 10 Hz, giving two-pass equivalence with one single pass. Patients scored the results as very good (n = 67), good (n = 25) and fair (n = 10). Mild but successfully resolved side-effects occurred in only four patients. The 2-month histology showed a good band of new collagen tightening the overlying healthy epidermis. Follow-up periods ranged from 1.5 to 2 years (mean +/- SD 1.76 +/- 0.33). This device at the above settings offers speedy resurfacing without compromising the quality of the procedure for the patient, and may well satisfy the basic requirements of laser skin resurfacing.


The interaction of bee melittin with lipid bilayer membranes.
The influence of melittin and the related 8-26 peptide on the stability and electrical properties of bilayer lipid membranes is reported. Melittin, unlike the 8-26 peptide, has a dramatic influence on lipid membranes, causing rupture at dilute concentrations. The circular dichroism of melittin demonstrated that under physiological conditions, in water, melittin is in extended conformation, which is enhanced in aqueous ethanol. However in 'membrane-like' conditions it is essentially alpha-helical. Secondary structure predictions were used to locate possible alpha-helical nucleation centres and a model of melittin was built according to these predictions. It is postulated that melittin causes a wedge effect in membranes.


Characterization of Burkholderia pseudomallei O antigens in different clinical strains.
O antigen is the major component of lipopolysaccharide LPS. The chemical structure of the O antigen determines the LPS serospecificity of the bacteria, and the diversity of O antigen is the basis for serotyping Burkholderia pseudomallei. In this study, structural elucidation of type B O antigen obtained from a clinical B. pseudomallei strain was conducted, and the effects of different types of LPS on macrophage differentiation were investigated. The O antigen was found to be composed of repeating units of [→4)-α-L-Rhap(1 → 4)-α-L-Rhap(1→2)-α-L-Rhap(1 → 2)-α-L-Rhap(1 → 3)-α-L-Rhap(1 → 3)-α-L-Rhap(1 → 4)-α-L-Rhap(1 → 6)-α-D-Galp(1→]n, where some of the →4)-α-L-Rhap(1 → units were substituted at O-3 by β-D-Xylp(1 → residues, and minor →3)-α-L-Rhap(1 → units were substituted at O-2 by β-D-Xylp(1 → residues. Meahwhile, the →6)-α-D-Galp(1 → units were substituted at O-3 by α-D-Galp(1 → residues. Furthermore, both type A and type B O antigens of B. pseudomallei could polarize macrophages toward the M1 phenotype, but the core oligosaccharides had no such activity. Therefore, we deduced that this polarization relies on the O antigen of LPS and might be related to the ability of B. pseudomallei to survive and replicate within macrophages. Thus, the characterization of different types of O antigen structural motifs is essential for further clarifying the persistence/survival mechanisms and inflammatory potential of B. pseudomallei.


Effects of flaxseed powder in improving non-alcoholic fatty liver by regulating gut microbiota-bile acids metabolic pathway through FXR/TGR5 mediating.
Non-alcoholic fatty liver disease (NAFLD) is gradually becoming one of the most common and health-endangering diseases. Flaxseed powder (FLA) is rich in α-linolenic acid, dietary fiber, lignans, and other active ingredients, which have lipid-lowering and anti-inflammatory effects. Here, we investigated whether the FLA improves host metabolism by gut bacteria modulation and further bile acid modulation in mice fed a high-fat diet. At the end of the experiment, we found that FLA can significantly reduce the body weight, body fat content, and serum TG, LDL-C, and TNF-α levels of mice, and improve liver steatosis. FLA intervention has a significant effect on preventing and regulating the gut flora disturbance caused by HFD. FLA intervention affects bile acid metabolism in the intestine and causes significant changes in functional bile acids, which can play a lipid-lowering and anti-inflammatory role by activating the intestinal Fxr- Fgfr4-Cyp7a1 and Tgr5-Tlr4-Tnfα pathways.


Automatic implantable cardioverter-defibrillator structural characteristics.
There are a number of equipment options and surgical techniques available for automatic implantable cardioverter-defibrillator implantation. The system can be successfully used even in problem cases where restrictions may be imposed because of physical build or the presence of other implanted devices. The sensing requirements and energy output of the units can be tailored to the exact needs of the particular patient. Battery life and device function are easily monitored periodically following implantation, making possible elective replacement of the pulse generator when the batteries become depleted.


Role of sex identification in perception of a social blunder.
4 groups of subjects observed an interaction between a male and a female on videotape: in one interaction the male blundered, in another the female blundered, and there were two control conditions in which the interaction was not followed by a blunder. Results indicated an effect of sex identification; the male blunderer was derogated most by male subjects (n = 34) and the female most by female subjects (n = 34).


Quantitative autoradiography of nicotinic receptors in large cryosections of human brain hemispheres.
In vitro quantitative autoradiography was used to visualize nicotinic receptors in large cryosections of human brain hemispheres. Sections 80 microns thick of human brain hemispheres were incubated with (-)-[3H]nicotine and the distribution of nicotinic receptors in human brain was studied. Increasing numbers of nicotinic binding sites were observed in the: hippocampus less than cortex less than cerebellum less than substantia nigra less than putamen less than periaqueductal gray. The cartography of nicotinic receptors in the normal human brain will hopefully be of use in the study of the alteration of these receptors in diseased brain.