Identification of Leg Chemosensory Genes and Sensilla in the Apolygus lucorum.
Apolygus lucorum (Hemiptera: Miridae), one of the main insect pests, causes severe damage in cotton and many other economic crops. As is well-known, legs play important roles in the chemoreception of insects. In this study, the putative chemosensory proteins in legs of A. lucorum involved in close or contact chemical communication of adult bugs were investigated using RNA transcriptome sequencing and qPCR methods. Transcriptome data of forelegs, middle legs and hind legs of adult bugs demonstrated that 20 odorant binding protein (OBP) genes, eight chemosensory protein (CSP) genes, one odorant receptor (OR) gene, one ionotropic receptor (IR) gene and one sensory neuron membrane protein (SNMP) gene were identified in legs of A. lucorum. Compared to the previous antennae transcriptome data, five CSPs, IR21a and SNMP2a were newly identified in legs. Results of qPCR analysis indicated that all these putative chemosensory genes were ubiquitously expressed in forelegs, middle legs and hind legs of bugs. Furthermore, four types of sensilla on legs of A. lucorum including sensilla trichodea (subtypes: long straight sensilla trichodea, Str1; long curved sensilla trichodea, Str2), sensilla chaetica (subtypes: sensilla chaetica 1, Sch1; sensilla chaetica 2, Sch2; and sensilla chaetica 3, Sch3), sensilla basiconca (subtypes: medium-long sensilla basiconca, Sba1; short sensilla basiconca, Sba2) and Böhm bristles (BB) were found using scanning electron microscopy. Additionally, the largest number of sensilla was observed on hind legs, while the forelegs had the smallest number of sensilla. Our data provide valuable insights into understanding the chemoreception of legs in A. lucorum.


Evaluation of neuroprotection by lithium and valproic acid against ouabain-induced cell damage.
The pathophysiology of manic-depression may be associated with dysregulation of ion homeostasis. Ouabain is a potent inhibitor of the sodium-potassium adenosine triphosphatase and has been purported to mimic abnormalities seen in acute mania. As manic episodes are believed to be neurotoxic and mood stabilizers have recently been implicated as neuroprotectants, it is of interest to determine if lithium and valproic acid antagonize ouabain-induced neurotoxicity. Human neuroblastoma SH-SY5Y cells were differentiated for 12 days then pretreated with lithium or valproic acid for 24 h and then challenged with a 10 microM ouabain insult. Cellular damage was assessed with lactate dehydrogenase (LDH) release, and apoptotic potential of ouabain was evaluated with DNA fragmentation. Ouabain significantly increased LDH release after 72 h of treatment. Lithium pretreatment at 1 mM diminished ouabain-induced LDH release. Valproic acid alone at 100 and 1000 micrograms/mL significantly increased LDH release from the cells. Furthermore, it significantly potentiated ouabain-induced LDH release. DNA fragmentation suggests that ouabain induces apoptosis. Lithium at the therapeutic level of 1 mM limits the extent of cellular damage caused by 10 microM ouabain in SH-SY5Y cells as measured by LDH release. Valproic acid alone at the therapeutic concentration of 100 micrograms/mL induces LDH release and does not prevent ouabain-induced LDH release.


Our Timing to Remove Peritoneal Catheter Dialysis After Kidney Transplant.
Patients on peritoneal dialysis treatment represent 15% of the global dialysis population. The major complication of peritoneal dialysis is catheter and peritoneal infection. Peritoneal dialysis patients who receive kidney transplants are at increased risk of infection because of immunosuppressive therapy. The purpose of this study is to show our ideal timing to remove peritoneal catheter after kidney transplant, which gives adequate security on renal function recovery and reduction of septic risk. We analyzed the outcomes of 65 patients on peritoneal dialysis who underwent kidney transplant between 2000 and 2016. In 61 cases there was an immediate graft functional recovery. In 4 cases there was a delayed graft function (DGF), and we performed a hemodialysis with temporary placement of a venous catheter. In all patients we removed peritoneal dialysis catheter 30 to 45 days after transplant. There has been 1 case of catheter infection, which was treated with antibiotic therapy. Our average time to remove the peritoneal dialysis catheter was shorter than times in previous studies, between the 30th and 45th postoperative day. In the 4 cases in which there has been a DGF, we performed hemodialysis treatment to avoid, in the immediate postoperative period, direct insults to the peritoneum by local dialysis procedures. Our experience show that the 30th to 45th postoperative day is a good time frame, better yet a good watershed between the safe removal of peritoneal catheter when patients have a stabilized renal function and the possibility of leaving it in situ, to resume peritoneal dialysis in case of persistent DGF.


Insulin sensitization by small molecules enhancing GLUT4 translocation.
Insulin resistance (IR) is the root cause of type II diabetes, yet no safe treatment is available to address it. Using a high throughput compatible assay that measures real-time translocation of the glucose transporter glucose transporter 4 (GLUT4), we identified small molecules that potentiate insulin action. In vivo, these insulin sensitizers improve insulin-stimulated GLUT4 translocation, glucose tolerance, and glucose uptake in a model of IR. Using proteomic and CRISPR-based approaches, we identified the targets of those compounds as Unc119 proteins and solved the structure of Unc119 bound to the insulin sensitizer. This study identifies compounds that have the potential to be developed into diabetes treatment and establishes Unc119 proteins as targets for improving insulin sensitivity.


Exome Sequencing and Identification of Phenocopies in Patients With Clinically Presumed Hereditary Nephropathies.
Hereditary nephropathies are clinically and genetically heterogeneous disorders. For some patients, the clinical phenotype corresponds to a specific hereditary disease but genetic testing reveals that the expected genotype is not present (phenocopy). The aim of this study was to evaluate the spectrum and frequency of phenocopies identified by using exome sequencing in a cohort of patients who were clinically suspected to have hereditary kidney disorders. Cross-sectional cohort study. 174 unrelated patients were recruited for exome sequencing and categorized into 7 disease groups according to their clinical presentation. They included autosomal dominant tubulointerstitial kidney disease, Alport syndrome, congenital anomalies of the kidney and urinary tract, ciliopathy, focal segmental glomerulosclerosis/steroid-resistant nephrotic syndrome, VACTERL association, and "other." A genetic diagnosis (either likely pathogenic or pathogenic variant according to the guidelines of the American College of Medical Genetics) was established using exome sequencing in 52 of 174 (30%) cases. A phenocopy was identified for 10 of the 52 exome sequencing-solved cases (19%), representing 6% of the total cohort. The most frequent phenocopies (n=5) were associated with genetic Alport syndrome presenting clinically as focal segmental glomerulosclerosis/steroid-resistant nephrotic syndrome. Strictly targeted gene panels (<25 kilobases) did not identify any of the phenocopy cases. The spectrum of described phenocopies is small. Selection bias may have altered the diagnostic yield within disease groups in our study population. The study cohort was predominantly of non-Finnish European descent, limiting generalizability. Certain hereditary kidney diseases cannot be diagnosed by using exome sequencing (eg, MUC1-autosomal dominant tubulointerstitial kidney disease). Phenocopies led to the recategorization of disease and altered clinical management. This study highlights that exome sequencing can detect otherwise occult genetic heterogeneity of kidney diseases.


Oxygen--insensitive nitrofuran reductases in Salmonella typhimurium TA100.
The present study demonstrated by DEAE-cellulose column chromatography that oxygen-insensitive nitrofuran reductases in Salmonella typhimurium TA100 consisted of at least two reductases, NADPH- and NAD(P)H-linked enzymes. The NADPH- and NADH-linked activities of the latter enzyme seemed to originate from a single enzyme, because both activities were similarly inactivated by heat and urea treatments, and also inhibited by dicumarol. On the other hand, the NADPH-linked enzyme was less sensitive to heat, urea and dicumarol. Furthermore, the study showed that the NAD(P)H-linked enzyme was a flavoenzyme which could be inactivated by dialysis against 1 M potassium bromide and reactivated by FMN.


Extensive feeding on sockeye salmon Oncorhynchus nerka smolts by bull trout Salvelinus confluentus during initial outmigration into a small, unregulated and inland British Columbia river.
Stomach contents were collected and analysed from 22 bull trout Salvelinus confluentus at the edge of the Chilko Lake and Chilko River in British Columbia, Canada, during spring outmigration of sockeye salmon Oncorhynchus nerka smolts. Twenty of the 22 (>90%) stomachs contained prey items, virtually all identifiable prey items were outmigrant O. nerka smolts and stomach contents represented a large portion (0·0-12·6%) of estimated S. confluentus mass. The results demonstrate nearly exclusive and intense feeding by S. confluentus on outmigrant smolts, and support recent telemetry observations of high disappearance rates of O. nerka smolts leaving large natural lake systems prior to entering high-order unregulated river systems.


Efficacy of Rhesus Theta-Defensin-1 in Experimental Models of Pseudomonas aeruginosa Lung Infection and Inflammation.
Chronic airway infection and inflammation contribute to the progressive loss of lung function and shortened survival of patients with cystic fibrosis (CF). Rhesus theta defensin-1 (RTD-1) is a macrocyclic host defense peptide with antimicrobial and immunomodulatory activities. Combined with favorable preclinical safety and peptide stability data, RTD-1 warrants investigation to determine its therapeutic potential for treatment of CF lung disease. We sought to evaluate the therapeutic potential of RTD-1 for CF airway infection and inflammation using in vitro, ex vivo, and in vivo models. We evaluated RTD-1's effects on basal and Pseudomonas aeruginosa-induced inflammation in CF sputum leukocytes and CF bronchial epithelial cells. Peptide stability was evaluated by incubation with CF sputum. Airway pharmacokinetics, safety, and tolerance studies were performed in naive mice. Aerosolized RTD-1 treatment effects were assessed by analyzing lung bacterial burdens and airway inflammation using an established model of chronic P. aeruginosa endobronchial infection in CF (ΔF508) mice. RTD-1 directly reduces metalloprotease activity, as well as inflammatory cytokine secretion from CF airway leukocyte and bronchial epithelial cells. Intrapulmonary safety, tolerability, and stability data support the aerosol administration route. RTD-1 reduced the bacterial lung burden, airway neutrophils, and inflammatory cytokines in CF mice with chronic P. aeruginosa lung infection. Collectively, these studies support further development of RTD-1 for treatment of CF airway disease.


Simple preparation of 3I-O-substituted beta-cyclodextrin derivatives using cinnamyl bromide.
[reaction: see text] A new method for preparation of 3(I)-O-substituted beta-cyclodextrin derivatives was developed. Cinnamyl bromide reacts with beta-cyclodextrin to form predominantly the 3(I)-O-cinnamyl derivative (30% isolated yield, >90% regioselectivity). After protection of the remaining cyclodextrin hydroxyls by acetylation, the cinnamyl group can be easily transformed to many other groups (exemplified by transformation to 3(I)-O-carboxymethyl derivative). Substitution pattern in singly modified CDs was unambiguously determined by a combination of 2D NMR techniques.


Relationship between the structural and functional changes of the photosynthetic apparatus during chloroplast-chromoplast transition in flower bud of Lilium longiflorum.
The relationship between the structural and functional changes of the photosynthetic apparatus in the flower bud of Lilium longiflorum during chloroplast-chromoplast transition was examined. Compared with green petals, there was a five-fold increase of the carotenoid content in yellow petals, whereas the chlorophyll content decreased five-fold. Absorption and emission fluorescence spectra of chromoplasts indicated that newly synthesized carotenoids were not associated with photosystem II (PSII) photochemistry. The maximum quantum yield in the remaining PSII reaction centers remained constant during the chromoplast formation, whereas the photosynthetic electron transport beyond PSII became inhibited, as indicated by a marked decrease of the O2 evolution capacity, of the photochemical quenching of chlorophyll-alpha fluorescence and of the operational quantum yield of photosynthetic electron transport. Deconvoluted fluorescence emission spectra indicated a more rapid degradation of photosystem I (PSI) complexes than of PSII during chromoplast formation. Compared with green petals, the spillover between PSII and PSI decreased by approximately 40% in yellow petals. Our results indicate that during chloroplast-chromoplast transition in the flower bud of L. longiflorum, PSII integrity was preserved longer than the rest of the photosynthetic apparatus.


HtrA3: a promising prognostic biomarker and therapeutic target for head and neck squamous cell carcinoma.
The dysregulation of the human high-temperature requirement A (HtrA) family of serine proteases is associated with many malignancies. However, there are few reports on HtrAs in head and neck squamous cell carcinoma (HNSCC). The aim of this study was to investigate the expression, prognostic value, and biological functions of HtrAs in HNSCC. The RNA-sequencing data and clinical data of HNSCC were downloaded from The Cancer Genome Atlas (TCGA) database. The GSE30784 and GSE31056 datasets from the Gene Expression Omnibus (GEO) database were used for further verification. This study explored the differential expression of HtrAs and assessed their potential impact on the prognosis of HNSCC patients using a survival module. Correlations between clinical characteristics and HtrA expression levels were then explored using a Wilcoxon rank sum test. A Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA) were performed using "clusterProfile" in the R software. A Pearson/Spearman correlation test was applied to analyze the relationship between HtrAs and immune infiltration level/checkpoint genes. Validation of HtrA expression levels were carried out by RT-PCR and western blot in human squamous carcinoma cell lines (Fadu and Cal-27) and human non-tumorigenic bronchial epithelium cells (BEAS-2B). Finally, through cell transfection, CCK-8, Ki-67 immunofluorescence, and flow cytometry assays, the effect of HtrA3 knockdown on the malignant biological behavior of HNSCC cells was explored. The gene expression levels of HtrAs were significantly upregulated and associated with patient age, TNM stage, clinical stage, and TP53 mutation status in the TCGA-HNSCC cohort. High expressions of HtrA1/3 were associated with shorter overall survival, shorter progress-free interval, and lower disease-specific survival in HNSCC. A nomogram for HtrAs was constructed and validated. HtrA-related genes were significantly enriched in the immune response and cell apoptosis pathway. In addition, the expression of HtrAs showed significant correlations with B cells, M cells, DC cell infiltration, and immune infiltration checkpoint (CD276, TNFRSF14). Validation of HtrA expression was carried out by RT-PCR and western blot. Results of in vitro experiments indicated that HtrA3 gene knockdown inhibits the proliferation of FaDu and Cal-27 cells while concurrently promoting apoptosis. HtrA3 shows significant potential as both a prognostic marker and a promising therapeutic target for HNSCC, highlighting its relevance and importance in future research and potential clinical applications.


Graphene-DNAzyme Junctions: A Platform for Direct Metal Ion Detection with Ultrahigh Sensitivity.
Many metal ions are present in biology and in the human body in trace amounts. Despite numerous efforts, metal sensors with ultrahigh sensitivity (< a few picomolar) are rarely achieved. Here, we describe a platform method that integrates a Cu2+-dependent DNAzyme into graphene-molecule junctions and its application for direct detection of paramagnetic Cu2+ with femtomolar sensitivity and high selectivity. Since DNAzymes specific for other metal ions can be obtained through in vitro selection, the method demonstrated here can be applied to the detection of a broad range of other metal ions.


Recent studies on the involvement of retinyl phosphate as a carrier of mannose in biological membranes.
Rat liver microsomes synthesized [14C]mannosylretinylphosphate and dolichyl [14C]mannosylphosphate from guanosinedisphosphate [14C]mannose, retinylphosphate and dolichylphosphate. Two distinct enzyme activities were shown to be responsible for the biosynthesis of the two mannolipids. A higher affinity mannosyl transferase (EA I), responsible for dolichylmannosylphosphate synthesis, displayed a Km for GDP-mannose of 1.7 microM; while a lower affinity enzyme (EA II), responsible for mannosylretinylphosphate synthesis, displayed a Km for GDP-mannose of 12.5 microM. These Km values were unaffected by the addition of either dolichylphosphate for EA II, or retinylphosphate for EA I. The same Km values were found before and after solubilization of the enzyme activity with 1% Triton X-100. Differential solubilization of EA I and EA II was demonstrated, utilizing different concentrations of Triton X-100. Triple-labeled mannosylretinylphosphate was prepared from [3H]retinylphosphate, retinyl[32P]phosphate and GDP-[14C]mannose from incubations containing rat liver microsomes. This compound was shown to donate [14C]mannose to endogenous acceptors of rat liver microsomes.


Minimally Invasive Open Surgical Approach and Outcomes for Carpal Tunnel Syndrome.
The most common peripheral neuropathy is carpal tunnel syndrome. The present study aims to describe our minimally invasive open surgical approach for carpal tunnel syndrome and evaluate surgical outcomes. We included 217 patients who were operated in our clinic for carpal tunnel syndrome by minimally invasive open surgical approach. Visual Analogue Scale and Functional Outcome Scale scores were obtained preoperative, postoperative at one month and three months to determine surgical outcomes. The mean age of the patients was 55.4±12.8 years (32 to 69), 175 (80.6%) were women and 42 (19.4%) were men. The assessment of carpal tunnel syndrome's etiology showed that 189 (%87.1%) of the cases were idiopathic, 19 (8.8%) had hypothyroidism, 5 (2.3%) had rheumatoid arthritis and 4 (1.8%) were due to pregnancy. The average improvement of VAS between preoperatively and late postoperatively was 5.41±1.05. The average improvement FOS was 17.44±3.06. They were statistically significant. The minimally invasive open surgical approach for carpal tunnel syndrome (an average of 1 cm skin incision) is performed with local anesthesia and successful surgical outcomes are achieved.


Bedside scoring procedure for the diagnosis of diabetic peripheral neuropathy in young patients with type 1 diabetes mellitus.
To test the applicability of a bedside scoring method for screening for diabetic peripheral neuropathy (DPN) in patients with type 1 diabetes mellitus (DM) in an ambulatory clinic. The prevalence of DPN was estimated and its risk factors identified. A total of 217 patients (102 males) with type 1 DM, median age 23.4 years (7.5-49 years) and median duration of DM 13.2 years (1-34 years) were evaluated for DPN using the bedside Neuropathy Disability Score (NDS). A score of 3-5 indicated mild DPN, 6-8 moderate DPN and 9-10 severe DPN. The presence of DPN was correlated with possible predictive factors. The NDS was reliable and highly reproducible. The overall prevalence of DPN was 17.1%: mild in 14.3%, moderate in 2.3%, and severe in 0.5% of patients. The prevalence and severity of DPN were significantly related to long-term glycemic control (p < 0.001), DM duration (p < 0.005), age (p = 0.005), and duration of pubertal DM duration (p = 0.03). The prevalence of DPN was significantly associated with the presence of retinopathy (p < 0.002) and overt proteinuria (p < 0.005). The NDS is a simple, reliable and reproducible screening method for use in the ambulatory clinic to identify the early signs of DPN, leading to early institution of intensive diabetes control measures and preventive foot care.


Fluorometric determination of N-nitroso-N-methylurea with nicotinamide and acetophenone.
A fluorometric method for the determination of N-nitroso-N-methylurea (NMU) has been developed. It is based on the N-methylation reaction of nicotinamide with NMU and a subsequent condensation reaction with acetophenone, followed by an acid treatment to form a fluorescent 2,7-naphthyridine derivative. This method enabled the determination of NMU in the range 0.05 - 2 nmol/200 microl with a relative standard deviation of ca. 3%. It was applied to the determination of NMU formed from a precursor N-methylurea (MU) under simulated gastric conditions containing nitrite and thiocyanate ions at pH 3.0 in the presence of fresh orange juice and milk. NMU was extracted by an Extrelut 20 column and then determined. The mean recoveries of NMU added to the simulated gastric juice containing water, orange juice and milk were 86.5, 85.1 and 69.8%, respectively. The amounts of NMU formed from MU were found to decrease to below 25% in the presence of orange juice and milk.


Key residues responsible for acyl carrier protein and beta-ketoacyl-acyl carrier protein reductase (FabG) interaction.
Fatty acid synthesis in bacteria is catalyzed by a set of individual enzymes collectively known as type II fatty-acid synthase. Each enzyme interacts with acyl carrier protein (ACP), which shuttles the pathway intermediates between the proteins. The type II enzymes do not possess primary sequence similarity that defines a common ACP-binding site, but rather are hypothesized to possess an electropositive/hydrophobic surface feature that interacts with the electronegative/hydrophobic residues along helix alpha2 of ACP (Zhang, Y.-M., Marrakchi, H., White, S. W., and Rock, C. O. (2003) J. Lipid Res. 44, 1-10). We tested this hypothesis by mutating two surface residues, Arg-129 and Arg-172, located in a hydrophobic patch adjacent to the active site entrance on beta-ketoacyl-ACP reductase (FabG). Enzymatic analysis showed that the mutant enzymes were compromised in their ability to utilize ACP thioester substrates but were fully active in assays with a substrate analog. Direct binding assays and competitive inhibition experiments showed that the FabG mutant proteins had reduced affinities for ACP. Chemical shift perturbation protein NMR experiments showed that FabG-ACP interactions occurred along the length of ACP helix alpha2 and extended into the adjacent loop-2 region to involve Ile-54. These data confirm a role for the highly conserved electronegative/hydrophobic residues along ACP helix alpha2 in recognizing a constellation of Arg residues embedded in a hydrophobic patch on the surface of its partner enzymes, and reveal a role for the loop-2 region in the conformational change associated with ACP binding. The specific FabG-ACP interactions involve the most conserved ACP residues, which accounts for the ability of ACPs and the type II proteins from different species to function interchangeably.


Epidermal growth factor receptor expression in follicular dendritic cells: a shared feature of follicular dendritic cell sarcoma and Castleman's disease.
The factors regulating the growth of follicular dendritic cell (FDC) sarcoma are currently unknown. Using cDNA microarray analysis, we found that the epidermal growth factor receptor (EGFR) is expressed in FDC sarcoma. We immunohistochemically examined the expression of EGFR in a larger series of FDC sarcomas and in nonneoplastic FDCs. This included 8 cases of FDC sarcoma, 12 cases of hyaline vascular Castleman's disease (CD), 5 cases of human herpesvirus 8 (HHV8)-positive plasma cell CD, 7 cases of HHV8-negative plasma cell CD, 13 cases of reactive lymph nodes, 3 cases of reactive tonsils, 10 cases of follicular lymphoma, 6 cases of nodular mantle cell lymphoma, and 6 cases of angioimmunoblastic T-cell lymphoma. EGFR was expressed in tumor cells in 7 of 8 cases (88%) of FDC sarcoma (strongly in 4 cases and moderately in 3 cases). The single EGFR-negative case had an anaplastic appearance and a more aggressive clinical behavior. EGFR was also expressed by FDC in all types of CD (strongly in 4 cases, moderately in 16 cases, and weakly in 4 cases). Immunostaining results were negative or only weakly positive for EGFR in FDC of reactive lymph nodes and tonsils, and in the FDC networks of follicular lymphoma, mantle cell lymphoma, and angioimmunoblastic lymphoma. The up-regulation of EGFR in FDC of CD was paralleled by an increase in EGFR expression in the surrounding perifollicular fibroblastic reticulum cells suggesting coordinate regulation. These findings identify a differentially expressed growth regulatory receptor common to both FDC sarcoma and CD, identifying a target for possible therapy in unresectable or refractory cases.


Successful treatment with trisodium phosphonoformate for primary cytomegalovirus infection after heart transplantation.
Primary cytomegalovirus infection in the patient who is receiving immunosuppression therapy is associated with a high morbidity and mortality. We report a patient who developed primary cytomegalovirus infection 37 days after heart transplantation with a rapidly deteriorating course. Treatment with the new antiviral drug trisodium phosphonoformate (Foscarnet-Astra) was initiated as a lifesaving measure with rapid, dramatic improvement in the patient's condition and subsequent recovery.


[LIMITATIONS OF QUESTIONNAIRE-BASED EVALUATION OF LOWER URINARY TRACT DYSFUNCTION IN ELDERLY PATIENTS].
(Purpose) Questionnaires are frequently used to evaluate subjective symptoms in clinical practice and research on lower urinary tract dysfunction. The usefulness and reliability of questionnaires such as the International Prostate Symptom Score (IPSS) and the Overactive Bladder Symptom Score (OABSS) are well known. However, elderly patients are often unable to fill out such questionnaires. There are no reports on the proportion of patients unable to complete these questionnaires and the background factors that make their use difficult. We conducted a prospective observational study to clarify these factors. (Materials and methods) Participants were 32 patients admitted to the rehabilitation ward of our hospital who were able to urinate on their own. The mean age was 82.9±6.9 years (65-97 years), and there were 11 men and 21 women. The main causes of hospitalization were orthopedic disease (17 cases), internal medical disease (9 cases), cerebrovascular disease (4 cases), and neurological disease (2 cases). The total score on the Functional Independence Measure (FIM) motor domains was used to evaluate patient motor function. The FIM cognitive domain total score and the Mini-Mental State Examination (MMSE) score were used to evaluate cognitive function. Patients were given Japanese versions of the IPSS, IPSS-Quality of Life, and OABSS, and asked to complete 12 questions by circling the responses. If they were unable to complete the questionnaire on their own, an occupational therapist assisted them for 10 minutes. Patients were divided into three groups according to their responses to the 12 items: self-completed, completed with assistance, and not completed even with assistance. The percentage in each group was determined. The number of questions that could not be answered by self-completion was defined as the number of missing questions. Correlations between the number of missing questions and age, FIM motor domain score, FIM cognitive domain score, and MMSE score were evaluated. We also performed univariate and multivariate analyses of patient background factors for two groups: patients who could not complete the questionnaire on their own and those who could complete the questionnaire on their own. Gender, age, medical history, FIM motor domain score, FIM cognitive domain score, and MMSE score were analyzed. Twenty-eight cases, excluding four cases with missing FIM and MMSE data, were examined. Based on the evaluation of the number of missing questions described above, cutoff values for age, FIM motor domains, FIM cognitive domains, and MMSE were set. Fisher's exact test and logistic regression analysis were performed. (Results) For the 12 questionnaire items, 21 patients (65.6%) were able to complete the questionnaire on their own, 6 patients (18.8%) were able to complete it with assistance, and 5 patients (15.6%) were not able to complete it even with assistance. Age, FIM motor domain score, FIM cognitive domain score, and MMSE score all showed significant correlations with the number of missing items, with correlation coefficients of 0.362 (p=0.0417), -0.435 (p=0.0183), -0.622 (p=0.000318), and -0.455 (p=0.0149), respectively. The univariate analysis showed that two background factors indicating cognitive decline prevented self-completion of the questionnaire: FIM cognitive domain score <21 points (p=0.0000518) and MMSE score <24 points (p=0.0377). Multivariate analysis showed that cognitive decline, as indicated by <21 points on FIM cognitive domains, (odds ratio 133, 95% confidence interval 7.29-2,430, p=0.000965) affected patients' inability to complete the questionnaire on their own.


The value of uncertainty.
The author discusses some of the characteristics of Roy Schafer's contributions to psychoanalysis that he finds most valuable, such as his openness to uncertainty, his anti-reductive view of analytic constructions, his unique formulation of the analyst's role, and his close attention to how the patient engenders particular emotional reactions in the analyst. The author also presents a clinical vignette illustrating the value of the analyst's tolerance of uncertainty in the face of the patient's push for interpretations, explanations, and reassurance.


Physical interaction between specific E2 and Hect E3 enzymes determines functional cooperativity.
The cellular protein E6AP functions as an E3 ubiquitin protein ligase in the E6-dependent ubiquitination of p53. E6AP is a member of a family of functionally related E3 proteins that share a conserved carboxyl-terminal region called the Hect domain. Although several different E2 ubiquitin-conjugating enzymes have been shown to function with E6AP in the E6-dependent ubiquitination of p53 in vitro, the E2s that cooperate with E6AP in the ubiquitination of its normal substrates are presently unknown. Moreover, the basis of functional cooperativity between specific E2 and Hect E3 proteins has not yet been determined. Here we report the cloning of a new human E2, designated UbcH8, that was identified in a two-hybrid screen through specific interaction with E6AP. We demonstrate that UbcH7, an E2 closely related to UbcH8, can also bind to E6AP. The region of E6AP involved in complex formation with UbcH8 and UbcH7 was mapped to its Hect domain. Furthermore, we show that UbcH5 and UbcH6, two highly homologous E2s that were deficient for interaction with E6AP, could associate efficiently with another Hect-E3 protein, RSP5. Finally, only the E6AP-interacting E2s could function in conjunction with E6AP in the ubiquitination of an E6 independent substrate of E6AP, whereas the noninteracting E2s could not. Taken together, these studies demonstrate for the first time complex formation between specific human E2s and the Hect domain family of E3 proteins and suggest that selective physical interaction between E2 and E3 enzymes forms the basis of specificity for functionally distinct E2:E3 combinations.


[Ethics committees in the Czech Republic].
The authors submit a report on the establishment and activities of new advisory organs which developed in hospitals and at medical faculties and bear the name of ethical commissions. Their task is to approve grants, advisory activities as well as organizing public discussions of ethical problems. The basis of the report are data obtained by means of questionnaires completed by 62 respondents. At present there are 20 ethical commissions in the Czech Republic, 19 local ones and one central one.


Aptamer directly evolved from live cells recognizes membrane bound immunoglobin heavy mu chain in Burkitt's lymphoma cells.
The identification of tumor related cell membrane protein targets is important in understanding tumor progression, the development of new diagnostic tools, and potentially for identifying new therapeutic targets. Here we present a novel strategy for identifying proteins that are altered in their expression levels in a diseased cell using cell specific aptamers. Using an intact viable B-cell Burkitt's lymphoma cell line (Ramos cells) as the target, we have selected aptamers that recognize cell membrane proteins with high affinity. Among the selected aptamers that showed different recognition patterns with different cell lines of leukemia, the aptamer TD05 showed binding with Ramos cells. By chemically modifying TD05 to covalently cross-link with its target on Ramos cells to capture and to enrich the target receptors using streptavidin coated magnetic beads followed by mass spectrometry, we were able to identify membrane bound immunoglobin heavy mu chain as the target for TD05 aptamer. Immunoglobin heavy mu chain is a major component of the B-cell antigen receptor, which is expressed in Burkitt's lymphoma cells. This study demonstrates that this two step strategy, the development of high quality aptamer probes and then the identification of their target proteins, can be used to discover new disease related potential markers and thus enhance tumor diagnosis and therapy. The aptamer based strategy will enable effective molecular elucidation of disease related biomarkers and other interesting molecules.


Relevant advances in bone lengthening research: a bibliometric analysis of the 100 most-cited articles published from 2001 to 2017.
This study aimed to assess the scientific production of bone lengthening research by identifying the most-cited papers. All articles including the term 'bone lengthening' published between 2001 and 2017 were retrieved through the Web of Science database. The 100 most-cited articles on bone lengthening included a total of 4244 citations, with 414 (9.7%) citations in 2017. There was an average of 249.6 citations per year. The articles predominantly addressed biomechanics and bone formation (38). Different surgical techniques, including intramedullary nail (14), Ilizarov (nine), intramedullary skeletal kinetic distractor (ISKD) (six), Taylor spatial frame (6), the PRECICE device (three), and lengthening and submuscular locking plate (three), were the second most-studied topic. Most studies were therapeutic (58), whereas 30 studies were experimental investigations using animal models. Among the clinical studies, case series were predominant (level of evidence IV) (57). This study presents the first bibliometric analysis of the most relevant articles on bone lengthening. The list is relatively comprehensive in terms of identifying the top issues in this field. However, the most influential clinical studies have a poor level of evidence, although a slight tendency toward a better level of evidence has been observed in more recent years.


Phosphatidylethanolamine-phosphatidylglycerol bilayer as a model of the inner bacterial membrane.
Phosphatidylethanolamine (PE) and phosphatidylglycerol (PG) are the main lipid components of the inner bacterial membrane. A computer model for such a membrane was built of palmitoyloleoyl PE (POPE) and palmitoyloleoyl PG (POPG) in the proportion 3:1, and sodium ions (Na+) to neutralize the net negative charge on each POPG (POPE-POPG bilayer). The bilayer was simulated for 25 ns. A final 10-ns trajectory fragment was used for analyses. In the bilayer interfacial region, POPEs and POPGs interact readily with one another via intermolecular hydrogen (H) bonds and water bridges. POPE is the main H-bond donor in either PEPE or PEPG H-bonds; PGPG H-bonds are rarely formed. Almost all POPEs are H-bonded and/or water bridged to either POPE or POPG but PE-PG links are favored. In effect, the atom packing in the near-the-interface regions of the bilayer core is tight. Na+ does not bind readily to lipids, and interlipid links via Na+ are not numerous. Although POPG and POPE comprise one bilayer, their bilayer properties differ. The average surface area per POPG is larger and the average vertical location of the POPG phosphate group is lower than those of POPE. Also, the alkyl chains of POPG are more ordered and less densely packed than the POPE chains. The main conclusion of this study is that in the PE-PG bilayer PE interacts more strongly with PG than with PE. This is a likely molecular-level event behind a regulating mechanism developed by the bacteria to control its membrane permeability and stability consisting in changes of the relative PG/PE concentration in the membrane.


Crystal structure of calcium-independent subtilisin BPN' with restored thermal stability folded without the prodomain.
The three-dimensional structure of a subtilisin BPN' construct that was produced and folded without its prodomain shows the tertiary structure is nearly identical to the wild-type enzyme and not a folding intermediate. The subtilisin BPN' variant, Sbt70, was cloned and expressed in Escherichia coli without the prodomain, the 77-residue N-terminal domain that catalyzes the folding of the enzyme into its native tertiary structure. Sbt70 has the high-affinity calcium-binding loop, residues 75 to 83, deleted. Such calcium-independent forms of subtilisin BPN' refold independently while retaining high levels of activity [Bryan et al., Biochemistry, 31:4937-4945, 1992]. Sbt70 has, in addition, seven stabilizing mutations, K43N, M50F, A73L, Q206V, Y217K, N218S, Q271E, and the active site serine has been replaced with alanine to prevent autolysis. The purified Sbt70 folded spontaneously without the prodomain and crystallized at room temperature. Crystals of Sbt70 belong to space group P2(1)2(1)2(1) with unit cell parameters a = 53.5 A, b = 60.3 A, and c = 83.4 A. Comparison of the refined structure with other high-resolution structures of subtilisin BPN' establishes that the conformation of Sbt70 is essentially the same as that previously determined for other calcium-independent forms and that of other wild-type subtilisin BPN' structures, all folded in the presence of the prodomain. These findings confirm the results of previous solution studies that showed subtilisin BPN' can be refolded into a native conformation without the presence of the prodomain [Bryan et al., Biochemistry 31:4937-4945, 1992]. The structure analysis also provides the first descriptions of four stabilizing mutations, K43N, A73L, Q206V, and Q271E, and provides details of the interaction between the enzyme and the Ala-Leu-Ala-Leu tetrapeptide found in the active-site cleft.


Chemical Composition, Antimicrobial and Antioxidant Activities of the Flower Volatile Oils of Fagopyrum esculentum, Fagopyrum tataricum and Fagopyrum Cymosum.
The purpose of this study was to investigate the chemical composition and biological activity of the volatile oils (VOs) from the flowers of three buckwheat species, Fagopyrum esculentum, Fagopyrum tataricum and Fagopyrum cymosum. The VOs were obtained from the fresh buckwheat flowers by hydrodistillation, and were analyzed for their chemical composition by gas chromatography-mass spectrometry (GC-MS). Nonanoic acid (7.58%), (E)-3-hexen-1-ol (6.52%), and benzothiazole (5.08%) were the major constituents among the 28 identified components which accounted for 92.89% of the total oil of F. esculentum. 2-Pentadecanone (18.61%), eugenol (17.18%), 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester (13.19%), and (E,E)-farnesylacetone (7.15%) were the major compounds among the 14 identified components which accounted for 88.48% of the total oil of F. tataricum. Eugenol (12.22%), (E)-3-hexen-1-yl acetate (8.03%), linalool oxide (7.47%), 1-hexanol (7.07%), and benzothiazole (6.72%) were the main compounds of the 20 identified components which accounted for 90.23% of the total oil of F. cymosum. The three VOs were screened to have broad spectrum antibacterial activity with minimum inhibitory concentration (MIC) values ranged from 100.0 μg/mL to 800.0 μg/mL against the tested bacteria, and their median inhibitory concentration (IC50) values were from 68.32 μg/mL to 452.32 μg/mL. Xanthomonas vesicatoria was the most sensitive bacterium. Moreover, the flower VOs of F. esculentum, F. tataricum and F. cymosum also exhibited noteworthy antioxidant capacity with the IC50 value of 354.15 μg/mL, 210.63 μg/mL, and 264.92 μg/mL for the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, and the value of 242.06 μg/mL, 184.13 μg/mL, and 206.11 μg/mL respectively for the β-carotene-linoleic bleaching test. These results suggested the volatile oils of buckwheat flowers could be potential resource of natural antimicrobial and antioxidant agents.


Idiopathic primary chylopericardium in a dog.
A 7-year-old spayed female Labrador Retriever was evaluated because of pericardial effusion. The dog had a history of decreased appetite and exercise intolerance of 3 days' duration. Thoracic radiography performed by the referring veterinarian revealed a large cardiac silhouette. Heart sounds were muffled. Echocardiographic findings were indicative of severe pericardial effusion with cardiac tamponade; no pleural effusion was identified. Pericardiocentesis yielded a considerable amount of chylous fluid. A diagnosis of chylopericardium in the absence of pleural effusion was made. Conservative management was not effective, and subtotal pericardectomy and thoracic duct ligation were recommended. Surgery was postponed by the owners for 25 days, at which time the dog had both chylopericardium and chylothorax. The dog underwent subtotal pericardectomy and thoracic duct ligation; to delineate the thoracic duct, intraoperative lymphangiography was performed by injection of a radiopaque contrast agent directly into a mesenteric lymph node and subsequent injection of methylene blue solution into another mesenteric lymph node. Surgical treatment resulted in complete resolution of the clinical signs and pleural effusion. To the authors' knowledge, this is the first report of the development of chylopericardium prior to development of chylothorax in a dog. Treatment with thoracic duct ligation and pericardectomy resulted in complete resolution of the effusion and clinical signs.


Analgesic narcotic antagonists. 1. 8 beta-Alkyl-, 8 beta-acyl-, and 8 beta-(tertiary alcohol)dihydrocodeinones and -dihydromorphinones.
Conjugate addition of lithium dialkyl cuprates to codeinone (3) gave as the major product a series of 8 beta-alkyldihydrocodeinones 4a-m. A low yield of the 8 alpha-isomer 6 was isolated in several cases. 8 beta-Acyldihydrocodeinones 10 were prepared by the addition of acyl carbanion equivalents (protected cyanohydrin method or lithium bis(alpha-ethoxyvinyl)cuprate) to 3 followed by hydrolysis. 8 beta-Acetyldihydrocodeine (12) was reacted with MeLi or n-BuLi to give tertiary alcohols 13, which were oxidized to target dihydrocodeinones 14. The 8 beta-substituted compounds with unsaturated (4c,f,m), branched (4d,g,i-k), or large straight-chain (4h,l) alkyl groups, as well as the acyl (10a-d) and tertiary alcohol (14a,b) derivatives, were less active than dihydrocodeinone (4n) in the mouse writhing and rat tail-flick analgesic assays. The analgesically active 8 beta-methyl (4a) and 8 beta-ethyl (4b) compounds were converted to N-(cyclopropylmethyl)- and N-(cyclobutylmethyl)dihydronorcodeinones (17 and 18) and -dihydronormorphinones (19 and 20). Some of these compounds had mixed agonist-antagonist profiles of action. One of these compounds, N-(cyclopropylmethyl)-8 beta-ethyldihydronorcodeinone (17b), has been selected for further study in man.


Nonconventional Therapeutic Protocol for Type III Dens Invaginatus.
This study discusses a nonconventional therapeutic protocol for type III dens invaginatus. This condition is a disorder of dental development, caused by the invagination of enamel into coronal and/or radicular dentin structure. This promotes several structural alterations within the dental organ, which offers challenges and difficulties to perform the endodontic treatment when needed. This article reports a clinical case where a conservative approach was adopted to preserve the invagi-nated tooth, and endodontic treatment was performed in the main necrotic canal. Following 21 years of observation, a complete root formation could be seen, with dental pulp preservation of the pulpotomized tooth. Considering the reported difficulties for the treatment of dens invaginatus, conservative measures, such as pulpotomy to preserve the remaining dental pulp may be an excellent alternative to allow less invasive procedures, thus avoiding endodontic surgery. This study discusses a nonconventional therapeutic protocol for type III dens invaginatus. A conservative approach adopted preserved the invaginated tooth, and root canal treatment was performed in the main necrotic canal. Following 21 years of observation, there was complete root formation, with dental pulp preservation of the pulpotomized tooth.


Changes in TGF-beta 1 levels in gingiva, crevicular fluid and serum associated with periodontal inflammation in humans and dogs.
Transforming growth factor-beta (TGF-beta) represents a family of polypeptide growth factors, involved in embryogenesis, inflammation, regulation of immune responses and wound healing. To determine whether TGF-beta contributes to the evolution of periodontal disease, we assayed TGF-beta levels in gingiva and crevicular fluid of patients with gingivitis and periodontitis. In parallel, TGF-beta was quantified in gingival fluid and serum of beagles with experimentally-induced periodontitis. Disease was monitored by several clinical parameters including Plaque Index, Gingival Index, probing depth, and epithelial attachment loss. Gingival tissues were obtained from 9 patients at the time of periodontal surgery, and gingival fluid samples were collected from an additional population of 10 periodontal patients. In 14 beagles, experimental periodontitis was induced and gingival fluids collected 6 months later. Fluid was collected by paper strips and volume measured by Periotron. Additionally, sera was collected before and 9 months after the ligature-induced periodontitis in 7 beagles. The levels of TGF-beta 1 were measured by ELISA. In the patients, a significantly higher concentration of TGF-beta 1 was observed both in the gingival tissues and fluid samples obtained from the sites with deeper periodontal pockets than in the less involved sites. In beagles, TGF-beta 1 levels measured in gingival fluid were elevated in moderate disease, declining in fluid samples obtained from the pockets during more advanced experimental periodontitis. Furthermore, with the progression of experimental periodontitis, a decrease in TGF-beta 1 occurred in the sera of the beagle dogs. These data suggest that TGF-beta 1 may play a rôle in the pathogenesis and diagnosis of periodontal disease, and that its actions can be further explored in an animal model.


An interdisciplinary community diagnosis experience in an undergraduate medical curriculum: development at Ghent University.
Since 2002, the medical curriculum at Ghent University has incorporated a community diagnosis exercise, teaming medical students with master of social work and social welfare studies students. The course focuses on the interaction between the individual and the community in matters of health and health care. During one week, small groups of students visit patients and their caregivers in six underserved urban neighborhoods, and they combine these experiences with public health data, to develop a community diagnosis. Local family physicians and social workers monitor sessions. The course requires students to design an intervention tackling one community health issue. At the end of the course, the students present their diagnoses and interventions to community workers and policy makers who provide feedback on the results. In the authors' experience, medical and social work students all value the joint learning experience. The occasional culture clash is an added value. The one-week course is very intensive for students, mentors, and cooperating organizations. Although students criticize time restraints, they feel that they reach the outlined objectives, and they rate the overall experience as very positive. The authors find that this interdisciplinary, community-oriented exercise allows students to appreciate health problems as they occur in society, giving them insight into the interaction of the local community with health and health care agencies. Combining public health data with experiences originating from a patient encounter mimics real-life primary care situations. This campus-community collaboration contributes to the social accountability of the university.


The effect of sesquiterpene lactones on the synthesis of nucleic acid in cultures of human lymphocytes stimulated by phytohemagglutinin.
The investigated sesquiterpene lactones--alatolide, eupatoriopicrine and hydroxyisonobiline, at the concentration 5 microgram/ml, resulted in a complete inhibition of 3H-uridine incorporation into RNA and 3H-thymidine into DNA in cultures of human lymphocytes stimulated by PHA. The blast and mitotic index was close or equal to 0 at this concentration of substances. Susceptibility to the investigated lactones was highest at the initial stage of lymphocyte culture and it gradually decreased with the progression of blastic transformation. At the stage of advanced RNA and DNA synthesis, an addition of the substances to cultures was without any inhibitory effect on the synthesis process.


Identifying genes and regulatory pathways associated with the scleractinian coral calcification process.
Reef building corals precipitate calcium carbonate as an exo-skeleton and provide substratum for prosperous marine life. Biomineralization of the coral's skeleton is a developmental process that occurs concurrently with other proliferation processes that control the animal extension and growth. The development of the animal body is regulated by large gene regulatory networks, which control the expression of gene sets that progressively generate developmental patterns in the animal body. In this study we have explored the gene expression profile and signaling pathways followed by the calcification process of a basal metazoan, the Red Sea scleractinian (stony) coral, Stylophora pistillata. When treated by seawater with high calcium concentrations (addition of 100 gm/L, added as CaCl2.2H2O), the coral increases its calcification rates and associated genes were up-regulated as a result, which were then identified. Gene expression was compared between corals treated with elevated and normal calcium concentrations. Calcification rate measurements and gene expression analysis by microarray RNA transcriptional profiling at two time-points (midday and night-time) revealed several genes common within mammalian gene regulatory networks. This study indicates that core genes of the Wnt and TGF-β/BMP signaling pathways may also play roles in development, growth, and biomineralization in early-diverging organisms such as corals.


Evaluation of the equations to estimate the glomerular filtration rate in kidney transplant recipients.
This study assessed the performance of three methods for estimating glomerular filtration rate (GFR) in kidney transplant patients: the Cockcroft-Gault formula, the modification of diet in renal disease (MDRD) method, and the four-variable modification of diet in renal disease (four-variable MDRD), both as an overall estimate and as related to clinical disease stage. We analyzed data from 136 renal transplant patients including 84 men in an overall age range of 28 to 76 years. Patients were categorized into three groups according to GFR as determined by the arithmetical mean of the last four creatinine clearance determinations after outlying values had been excluded: group 1, estimated GFR of <30 mL/min (n = 26); group 2, estimated GFR of 30 to 60 mL/min (n = 63);, and group 3, estimated GFR >60 mL/min (n = 33). Fourteen patients were excluded from the analysis because of a high variability between their creatinine clearance determinations. Estimated GFRs using the Cockroft-Gault, MDRD, and four-variable MDRD formulae were compared with GFRs as measured by creatinine clearance. Statistically significant correlations were observed for all three formulae for the overall series and for individual clinical groups. Hence, we concluded that all equations had a similar capacity to predict the GFR. In addition, because of the clear, significant correlation between the MDRD and the four-variable MDRD (r = .992; P = .0001), we believe that the four-variable MDRD can substitute for the MDRD for clinical purposes.


Channel noise effects on first spike latency of a stochastic Hodgkin-Huxley neuron.
While it is widely accepted that information is encoded in neurons via action potentials or spikes, it is far less understood what specific features of spiking contain encoded information. Experimental evidence has suggested that the timing of the first spike may be an energy-efficient coding mechanism that contains more neural information than subsequent spikes. Therefore, the biophysical features of neurons that underlie response latency are of considerable interest. Here we examine the effects of channel noise on the first spike latency of a Hodgkin-Huxley neuron receiving random input from many other neurons. Because the principal feature of a Hodgkin-Huxley neuron is the stochastic opening and closing of channels, the fluctuations in the number of open channels lead to fluctuations in the membrane voltage and modify the timing of the first spike. Our results show that when a neuron has a larger number of channels, (i) the occurrence of the first spike is delayed and (ii) the variation in the first spike timing is greater. We also show that the mean, median, and interquartile range of first spike latency can be accurately predicted from a simple linear regression by knowing only the number of channels in the neuron and the rate at which presynaptic neurons fire, but the standard deviation (i.e., neuronal jitter) cannot be predicted using only this information. We then compare our results to another commonly used stochastic Hodgkin-Huxley model and show that the more commonly used model overstates the first spike latency but can predict the standard deviation of first spike latencies accurately. We end by suggesting a more suitable definition for the neuronal jitter based upon our simulations and comparison of the two models.


AIE Supramolecular Assembly with FRET Effect for Visualizing Drug Delivery.
Here, we constructed a nanostructured pH/redox dual-responsive supramolecular drug carrier with both aggregation-induced emission (AIE) and Forster resonance energy transfer (FRET) effects, which enabled selective drug release and monitoring drug delivery and release processes. Taking the hyperbranched polyamide amine (H-PAMAM) with intrinsic AIE effects as the core, poly(ethylene glycol) (PEG) was bridged on its periphery by dithiodipropionic acid. Then, through the host-guest interaction of PEG and α-cyclodextrin, the supramolecular nanoparticles with AIE effects were constructed to load the anticancer drug doxorubicin (DOX). The supramolecular assembly has sufficiently large DOX loading due to the abundant cavities formed by branched structures. The hyperbranched core H-PAMAM has strong fluorescence, and the dynamic track of drug carriers and the dynamic drug release process can be monitored by the AIE and FRET effects between H-PAMAM and DOX, respectively. Furthermore, the introduction of disulfide bonds and the pH sensitivity of H-PAMAM enable the achievement of rapid selective release of loaded DOX at the tumor while remaining stable under normal physiological conditions. In vitro cytotoxicity indicates that the drug-loaded supramolecular assembly has a good therapeutic effect on cancer. In addition, the H-PAMAM core is different from the traditional AIE functional group, which has no conjugated structure, such as a benzene ring, thereby providing better biocompatibility. This technology will have broad applications as a new drug delivery system.


Influence of Educational Intervention on Awareness and Attitude of Nursing Students Towards Adverse Drug Reaction Reporting During Coronavirus Disease 2019 (COVID-19) Pandemic: A Quasi-Experimental Study.
Under-reporting of adverse drug reactions (ADRs) is a major concern of the pharmacovigilance program of India and has deteriorated further with the Coronavirus Disease 2019 (COVID-19) pandemic. Imparting the knowledge of pharmacovigilance among nursing students would significantly enhance patient care. Hence, this study was framed to evaluate the effect of an educational intervention on awareness and attitude of ADR reporting among nursing students during the COVID-19 pandemic. A quasi-experimental pretest-post-test study was conducted among nursing students of a tertiary care hospital in South India. The students were asked to fill out a validated questionnaire to assess Knowledge, Attitude, and Practice on ADR reporting, before and after the educational intervention. Knowledge and attitude results were analysed using Mc Nemar's test. A total of 135 nursing students attended the educational intervention and completed pretest and post-test questionnaires. Around 130 (96.3%) and 107 (79.3%) students were aware of pharmacovigilance and ADR monitoring center following the intervention. Median (interquartile range) of knowledge scores before and after the intervention were 3 (2-3) and 5 (3-7), respectively. Overall knowledge score of ADR reporting significantly improved after the intervention. Sixty two (45.9%) students agreed that ADR reporting would enhance patient safety. ADR reporting was practised by 52 (38.5%) participants. Nearly, 107 (79.3%) students were interested to participate in similar pharmacovigilance training programs. Educational intervention in pharmacovigilance improved the cognitive and affective domains of pharmacovigilance in nursing students. Periodic training programs are essential for sustained behavioural change in students.


In vitro evaluation of the antimicrobial activity of antiseptics against clinical Acinetobacter baumannii strains isolated from combat wounds.
Healthcare-associated infections (HCAIs) are among the most prominent medical problems worldwide. In the context of increasing antibiotic resistance globally, the use of antiseptics as the main active agent and potentiator of antibiotics for the treatment of purulent-inflammatory complications of traumatic wounds, burns, and surgical wounds can be considered to tackle opportunistic infections and their prevention during war. This study presents a comparative investigation of the antimicrobial efficacy of antiseptics used for surgical antisepsis and antiseptic treatment of skin, mucous membranes, and wounds against multidrug-resistant clinical isolates of Acinetobacter baumannii as a wound pathogen of critical priority (according to the WHO). It was found that strains of A. baumannii, which have natural and acquired resistance to antimicrobial drugs, remain susceptible to modern antiseptics. Antiseptic drugs based on decamethoxine, chlorhexidine, octenidine, polyhexanide, and povidone-iodine 10% and 2% provide effective bactericidal activity against A. baumannii within the working concentrations of these drugs. Chlorhexidine and decamethoxine can inhibit biofilm formation by A. baumannii cells. In terms of bactericidal properties and biofilm formation inhibition, chlorhexidine and decamethoxine are the most effective of all tested antiseptics.


Left ventricular deformation in patients with lymphedema before and after the use of medical compression stockings-detailed analysis from the three-dimensional speckle-tracking echocardiographic MAGYAR-Path Study.
Lymphedema (LE) is a chronic condition, which refers to tissue swelling due to excess interstitial fluid accumulation or impaired lymphatic conduit. Theoretically, lymphedema-related fluid retention could affect left ventricular (LV) mechanics, which could be detailed by recent three-dimensional speckle-tracking echocardiography. Therefore, it was purposed to examine LV strains in LE patients before and one hour after the use of medical compression stockings and to compare findings to those of matched normal subjects. The study comprised 26 cases with lymphedema, however, 4 cases had to be excluded due to inferior quality of images. Their results were compared to 27 age- and gender-matched healthy controls. Global LV circumferential and area strains and mean segmental LV circumferential strain were increased in lymphedema patients before the use of medical compression stockings as compared to controls. One hour after the use of medical compression stockings, no global and mean segmental LV strain showed significant impairment or improvement, but tendentious reduction was seen in LV circumferential strain. With LV segmental analysis, midventricular LV radial, circumferential and area strains proved to be significantly increased, while basal LV longitudinal strain and midventricular LV three-dimensional strain were decreased as compared to controls. No changes in regional LV strains could be detected after one-hour medical compression stockings use as compared to data collected at rest. Increased global LV circumferential strain is seen in lymphedema. With using medical compression stockings, LV deformation parameters change towards the normal range emphasizing their importance on cardiac function.


Evolutionary perspective on the origin of Haitian cholera outbreak strain.
Cholera epidemic has not been reported in Haiti for at least 100 years, although cholera has been present in Latin America since 1991. Surprisingly, the recent cholera epidemic in Haiti (October 2010) recorded more than 250,000 cases and 4000 deaths in the first 6 months and became one of the most explosive and deadly cholera outbreak in recent history. In the present study, we conducted genomic analyses of pathogenicity islands of three Haitian Vibrio cholerae strains and compared them with nine different V. cholerae O1 El Tor genomes. Although CIRS101 is evolutionarily most similar to the Haitian strains, our study also provides some important differences in the genetic organization of pathogenicity islands of Haitian strains with CIRS101. Evolutionary analysis suggests that unusual functional constraints have been imposed on the Haitian strains and we hypothesize that amino acid substitution is more deleterious in Haitian strains than in nonHaitian strains.


Electroacupuncture for Functional Constipation: A Multicenter, Randomized, Control Trial.
Background and Aim. To investigate the efficacy and safety of electroacupuncture (EA) with different current intensities for functional constipation (FC) and to assess whether the effects of EA with different current intensities are superior to the mosapride. Methods. Patients with FC were randomly divided into low current intensity group (LCI), high current intensity group (HCI), and mosapride group (MC). The primary outcome was three or more spontaneous bowel movements (SBMs) per week and an increase of one or more SBMs from baseline during at least 3 of the 4 weeks. Results. The primary outcome was reached by 53.45%, 66.15%, and 52.24% of the patients who received LCI, HCI, and mosapride, respectively. EA can significantly improve the weekly SBMs and stool consistency and reduce straining severity (p < 0.0001, all). HCI improved the quality of life better than mosapride (p < 0.05) and reduced the proportion of severe constipation more than LCI and mosapride (p < 0.05, both). Conclusions. EA is effective and safe at both current intensities for FC; therapeutic effects of LCI and HCI are not superior to mosapride. EA is superior to mosapride in improving patients' life quality and satisfaction level of treatment; EA has fewer adverse events than mosapride.


Motherhood and genetic screening: a personal perspective.
According to the medical profession the direction and scope of reproductive services such as IVF and pre-natal screening are based on solid evidence; the evidence indicates these are effective and safe services. Moreover, women want them. As a consequence these services are usually presented to the wider community in a positive light with images of 'successful' birth outcomes showcasing the importance of their work. Unsurprisingly this has lead to women being expected to take control - from timing a pregnancy to choosing one particular pregnancy over another - they are to improve their lives and the health of their offspring. But are these developments all 'good' news? Is it safe to assume the push to achieve better birth outcomes and the concomitant use of prenatal testing automatically improves lives? Could it be the issues raised are causing some lives to become harder? How meaningful, for example, are tests such as amniocentesis and CVS? As the mother of a child with Down syndrome I believe it is important for myself and other women in similar situations to share their lived experience. Perhaps we can illuminate some of the more complex and troubling issues these technological advances have the capacity to create - not only for ourselves - but for all women.


Melt Electrospinning Writing of Three-dimensional Poly(ε-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications.
This tutorial reflects on the fundamental principles and guidelines for electrospinning writing with polymer melts, an additive manufacturing technology with great potential for biomedical applications. The technique facilitates the direct deposition of biocompatible polymer fibers to fabricate well-ordered scaffolds in the sub-micron to micro scale range. The establishment of a stable, viscoelastic, polymer jet between a spinneret and a collector is achieved using an applied voltage and can be direct-written. A significant benefit of a typical porous scaffold is a high surface-to-volume ratio which provides increased effective adhesion sites for cell attachment and growth. Controlling the printing process by fine-tuning the system parameters enables high reproducibility in the quality of the printed scaffolds. It also provides a flexible manufacturing platform for users to tailor the morphological structures of the scaffolds to their specific requirements. For this purpose, we present a protocol to obtain different fiber diameters using melt electrospinning writing (MEW) with a guided amendment of the parameters, including flow rate, voltage and collection speed. Furthermore, we demonstrate how to optimize the jet, discuss often experienced technical challenges, explain troubleshooting techniques and showcase a wide range of printable scaffold architectures.


Suprathreshold contrast sensitivity in experimentally strabismic monkeys.
Human strabismic amblyopes show deficits in spatial vision that can be revealed in a variety of visual tasks. In particular, they show a reduced sensitivity to contrast for a wide range of spatial frequencies. The ability of strabismic amblyopes to process contrast information at levels well above detection threshold is less well understood and somewhat controversial. In the course of investigating the neural basis of strabismic amblyopia we studied contrast processing both at and above detection threshold in experimentally strabismic monkeys (Macaca nemestrina). First we trained them to perform a contrast detection task and measured their contrast sensitivity for a wide range of spatial frequencies. Then we trained them to discriminate between two gratings that were identical except for their contrast. We show that these monkeys exhibit deficits in both tasks. The deficits in the contrast discrimination task cannot be solely attributed to their deficit at threshold.


[Treatment of a pregnant patient with bilateral traumatic bimalleolar ankle fracture: a case report].
The incidence of ankle fractures has been increasing among all fractures due to skeletal traumas. Bimalleolar fractures account for 25% of all ankle fractures. Bilateral traumatic bimalleolar ankle fracture is a very rare condition. In such a clinical presentation, pregnancy managed with cesarean procedure complicates the situation due to increased risk of venous thromboembolism and fatal pulmonary embolism. In this article, we report a 25-year-old female case within an eight-month pregnancy period who had bilateral bimalleolar ankle fracture due to an accident in a motor-vehicle and the clinical and radiological results of surgical treatment were discussed in the light of current literature data.


A stereoselective cobalt-containing nitrile hydratase.
Nitrile hydratase from Pseudomonas putida NRRL-18668 has been purified and characterized. The purified enzyme catalyzes the hydration of 2(S)-(4'-chlorophenyl)-3-methylbutyronitrile at least fifty times faster than that of 2(R)-(4'-chlorophenyl)-3-methylbutyronitrile. This enzyme is a member of the class of nitrile hydratase that contains cobalt. Visible absorption and CD spectra suggest the cobalt exists as a non-corrin low-spin Co3+ ion in a tetragonally-distorted octahedral ligand field. Chemical reduction of the native enzyme results in a species with the EPR signature of a low-spin Co2+ complex. Like the other cobalt-containing nitrile hydratases, this enzyme is relatively stable, maintaining its activity below 35 degrees C, and it shows a broad activity optimum between pH 7.2 and 7.8. The structural genes for this enzyme have been cloned and sequenced. The deduced amino acid sequences for the alpha and beta subunits show 48-63% and 35-41% homology, respectively, to other sequenced nitrile hydratases. In particular, the cysteine residues in the alpha subunit that have been suggested to coordinate the metal ion in the iron-containing nitrile hydratases [Brennan, B. A., Cummings, J. G., Chase, D. B., Turner, I. M., Jr., & Nelson, M. J. (1996) Biochemistry 35, 10068-10077] are conserved in this enzyme, suggesting that this nitrile hydratase, like the enzyme from Rhodococcus rhodochrous J1, is a member of a newly described class of metalloenzymes with Co3+-thiolate ligation [Brennan, B. A., Alms, G., Nelson, M. J., Durney, L. T., & Scarrow, R. C. (1996) J. Am. Chem. Soc. 118, 9194-9195].


Investigation on the Inhibitory Effect of Methotrexate on Rheumatoid Synovitis via the TLR4-NF-κB Pathway in a Rat Model.
Rheumatoid arthritis (RA) is a rheumatoid immune system disease characterized by joint inflammation, resulting in synovial hyperplasia, articular cartilage damage or distortion, and extra-articular involvement. The morbidity is higher and the treatments are not effective in clinical, and also no unified to the pathogenesis of such diseases. The aim of this paper is to establish a rat model of rheumatoid synovitis and observe the inhibitory effect of methotrexate on this disease. A total of 100 SD rats are selected and randomly divided into 5 groups, with 20 rats in each group. The cold and damp factors of rheumatoid arthritis are induced by cold water and the arthritis score is used to verify the model. ELISA is used to measure the protein expression of Toll-like Receptor 4 (TLR4), Nuclear Factor kappa-B (NF-κB) and inflammation-related factors, and SPSS25.0 is used for statistical analysis. The results show that there is no significant difference in inflammatory scores among the four groups except the control group. However, after 3 months of intervention, the inflammatory scores in the methotrexate groups are significantly lower than those in the model group, and in the methotrexate group, the higher the dose, the lower the inflammatory scores. The experimental results show that the messenger ribonucleic acid (mRNA) and protein expressions of TLR4 and NF-κB from high to low are in the order of model group > low dose > middle dose > high dose > control group, and the expression trend of inflammation-related factors is the same as mentioned above. These results indicate that methotrexate can repair rheumatoid synovitis by inhibiting the inflammatory signaling pathway TLR4-NF-κB.


Medical comorbidities increase the rate of surgical site infection in primary Achilles tendon repair.
To assess the effects of medical comorbidities on the incidence of surgical site infection following primary Achilles tendon repair. A secondary aim was to assess the effects of specific medical comorbidities on the cost and extent of healthcare utilization related to surgical site infection following primary Achilles tendon repair. 24,269 patients undergoing primary Achilles tendon repair between 2005 and 2012 were examined. Current Procedural Terminology codes for primary Achilles tendon repair, and incision and drainage were used to search for and compile patient data from the United Healthcare Orthopedic and Medicare databases. Primary outcome measures regarding surgical site infection following primary Achilles tendon repair included the rate of occurrence, cost, and duration of treatment. Patients with one or more preexisting medical comorbidities at the time of surgery had an increased rate of surgical site infection compared to those without. Diabetes and vascular complications were associated with the highest surgical site infection rates. The rate of surgical incision and drainage was higher in patients with cardiac arrhythmias and uncomplicated hypertension. The presence of a medical comorbidity significantly increased the cost and duration of surgical site infection treatment. Medical comorbidities can complicate the postoperative course for patients undergoing Achilles tendon repair, which increases the cost of care and duration of treatment. A better understanding of the relationship between each medical comorbidity and surgical site infections following Achilles tendon repair may be ascertained with additional prospective studies, thus, allowing for a more accurate evaluation and stratification of surgical candidates to improve patient outcomes. Retrospective cohort study, Level III.


Complete Tri-Axis Magnetometer Calibration with a Gyro Auxiliary.
Magnetometers combined with inertial sensors are widely used for orientation estimation, and calibrations are necessary to achieve high accuracy. This paper presents a complete tri-axis magnetometer calibration algorithm with a gyro auxiliary. The magnetic distortions and sensor errors, including the misalignment error between the magnetometer and assembled platform, are compensated after calibration. With the gyro auxiliary, the magnetometer linear interpolation outputs are calculated, and the error parameters are evaluated under linear operations of magnetometer interpolation outputs. The simulation and experiment are performed to illustrate the efficiency of the algorithm. After calibration, the heading errors calculated by magnetometers are reduced to 0.5° (1σ). This calibration algorithm can also be applied to tri-axis accelerometers whose error model is similar to tri-axis magnetometers.


Management of possible stroke by laypeople in pre-hospital setting: who should we train?
The pre-hospital management of a possible stroke is fundamental for the transport of patients to the correct HUB facility; thus, they must be transported to the Emergency Department (ED) by EMS vehicles. Our study aims to analyze the factors correlated with a higher probability of accessing the ED through the EMS in this event. This is a retrospective observational study. All accesses in the 120 EDs of the Lombardy region, with a diagnosis of discharge whose symptoms could resemble CPSS, were analyzed between 1 January 2019 and 31 December 2019. We identified an increased probability of using the EMS vehicles of 0,05% (I.C. 95%: 0.04% - 0.06%; p<0.0001) for each additional year of age, considering patients aged 20 to 100 years and the percentage was significantly higher in the female population (58% vs 49%; p<0.001). Moreover, we calculated that the incidence of stroke was approximately 140 cases per 100,000 inhabitants. Only half of the citizens in the Lombardy region use the EMS in case of suspicion of stroke; further information campaigns are essential to educate citizens. Information strategies should be directed especially at men between 30 and 59 years old.


A report on an acute, in-hours, outpatient review clinic with ultrasonography facilities for the early evaluation of general surgical patients.
Introduction In 2013 our hospital introduced an in-hours, consultant-led, outpatient acute surgical clinic (ASC) for emergency general surgical patients. In 2014 this clinic was equipped with a dedicated ultrasonography service. This prospective cohort study evaluated this service before and after the introduction of ultrasonography facilities. Methods Data were recorded prospectively for all patients attending the clinic during 2013 and 2014. The primary outcome was patient destination (whether there was follow-up/admission) after clinic attendance. Results The ASC reviewed patients with a wide age range and array of general surgical complaints. In 2013, 186 patients attended the ASC. After the introduction of the ultrasonography service in 2014, 304 patients attended. In 2014, there was a reduction in the proportion of patients admitted to hospital from the clinic (18.3% vs 8.9%, p=0.002). However, the proportion of patients discharged after ASC review remained comparable with 2013 (30.1% in 2013 vs 38.8% in 2014, p=0.051). The proportion of patients undergoing computed tomography (CT) scans also fell (14.0% vs 4.9%, p<0.001). Conclusions The ASC assessed a wide array of general surgical complaints. Only a small proportion required hospital admission. The introduction of an ultrasonography service was associated with a further reduction in admission rates and computed tomography.


Leucine-nicotinic acid synergy stimulates AMPK/Sirt1 signaling and regulates lipid metabolism and lifespan in Caenorhabditis elegans, and hyperlipidemia and atherosclerosis in mice.
Nicotinic acid (NA), a lipid-lowering drug, serves as a source of NAD+, the cofactor for Sirt1. Leucine (Leu) stimulates the AMPK/Sirt1 axis and amplifies the effects of other AMPK/Sirt1 activating compounds. Therefore, we tested the interactive effects of leucine and low dose NA on AMPK/Sirt1 signaling and downstream effects of lipid metabolism in cell culture, C. elegans and mice. LDL-receptor knockout mice were fed an atherogenic Western diet supplemented with leucine (24 g/kg diet) and sub-therapeutic NA combinations (50 mg/kg diet and 250 mg/kg diet) or low therapeutic NA (1000 mg/kg diet) for 8 weeks to evaluate markers of hyperlipidemia and atherosclerosis. NA-Leu increased P-AMPK and Sirt1 in adipocytes and myotubes. In C. elegans, NA-Leu increased P-AMPK and DAF-16 (FOXO), reduced lipid accumulation and increased median survival under mild oxidative stress conditions. In the mice, NA-Leu reduced total cholesterol, cholesterol esters, plasma triglycerides, atherosclerotic lesion size, lipid area, and aortic macrophage infiltration, similar to the therapeutic NA dose. Leu amplifies the effects of NA on lipid metabolism, hyperlipidemia and atherosclerosis in mice, at least in part by activation of the AMPK/Sirt1 axis. This combination may be a potential therapeutic alternative for hyperlipidemia and atherosclerosis.


Interpreting trends in alcohol consumption and alcohol related damage.
Some theoretical and methodological issues in the study of trends in alcohol related problems are discussed. Special attention is given to the aggregation problem, the time-lag problem, the fact that other etiological factors tend to change simultaneously with alcohol consumption and the fact that changes in drinking style must be taken into consideration in addition to changes in amounts consumed.


Protease-Activation of Fc-Masked Therapeutic Antibodies to Alleviate Off-Tumor Cytotoxicity.
The interaction of the Fc region of therapeutic antibodies and antibody-drug conjugates with Fcγ receptors (FcγRs) can lead to unpredictable and severe side effects. Over the last decades several strategies have been developed to overcome this drawback, including extensive Fc- and glycoengineering and antibody isotype switching. However, these approaches result in permanently Fc-silenced antibody derivates which partially or completely lack antibody-mediated effector functions. Nevertheless, for a majority of antibody-based drugs, Fc-mediated effector functions, like antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP) as well as complement-dependent cytotoxicity (CDC), represent the most substantial modes of action. We argued that a new strategy combining the beneficial properties of Fc-silencing and controlled activation of effector functions can pave the way to potent antibody therapeutics, reducing the FcγRs-mediated off-target toxicity. We present a novel Fc-tamed antibody format, where the FcγR-binding sites of antibodies are blocked by anti-isotypic masking units, hindering the association of FcγR and complement component 1 (c1q) to the Fc domain. The masking units were genetically fused to trastuzumab, including a protease-addressable peptide-liker. Our Fc-tamed antibodies demonstrated completely abolished interaction to soluble high-affinity Fcγ-Receptor I and c1q. In reporter cell-based ADCC assays, our Fc-tamed antibodies exhibited a 2,700 to 7,100-fold reduction in activation, compared to trastuzumab. Upon demasking by a tumor-associated protease, the Fc-activated antibodies demonstrated restored FcγR-binding, c1q-binding and the ability to induce potent ADCC activation. Furthermore, cell killing assays using donor-derived NK cells were performed to validate the functionality of the Fc-tamed antibody variants. To our knowledge, this approach represents the first non-permanently Fc-silenced antibody, which can be re-activated by a tumor-associated protease, eventually extending the field of novel antibody formats.


Hierarchical nitrogen-doped flowerlike ZnO nanostructure and its multifunctional environmental applications.
Hierarchical nitrogen-doped ZnO flowerlike nanostructures were synthesized on a large scale. These nanostructures were characterized by FESEM, HRTEM, XRD, FTIR, XPS, and TGA, and their suitability for multifunctional environmental applications was investigated. The experimental results demonstrated that the hierarchical N-doped ZnO flowerlike nanostructure enhances the photodegradation of methyl blue (MB) and acid orange 7 (AO7) by presenting a large specific surface area and high light utilization rate, inhibits the growth of bacteria without light irradiation, and increases the permeate flux when used in a membrane filtration system. These advantages of the hierarchical N-doped ZnO flowerlike nanostructure brings benefits to the environmental application fields.


A Long-Term Study Evaluating the Effects of Nicorandil Treatment on Duchenne Muscular Dystrophy-Associated Cardiomyopathy in mdx Mice.
Duchenne muscular dystrophy (DMD) is a neuromuscular disease caused by dystrophin gene mutations affecting striated muscle. Due to advances in skeletal muscle treatment, cardiomyopathy has emerged as a leading cause of death. Previously, nicorandil, a drug with antioxidant and nitrate-like properties, ameliorated cardiac damage and improved cardiac function in young, injured mdx mice. Nicorandil mitigated damage by stimulating antioxidant activity and limiting pro-oxidant expression. Here, we examined whether nicorandil was similarly cardioprotective in aged mdx mice. Nicorandil (6 mg/kg) was given over 15 months. Echocardiography of mdx mice showed some functional defects at 12 months compared to wild-type (WT) mice, but not at 15 months. Disease manifestation was evident in mdx mice via treadmill assays and survival, but not open field and grip strength assays. Cardiac levels of SOD2 and NOX4 were decreased in mdx vs. WT. Nicorandil increased survival in mdx but did not alter cardiac function, fibrosis, diaphragm function or muscle fatigue. In contrast to our prior work in young, injured mdx mice, nicorandil did not exert cardioprotective effects in 15 month aged mdx mice. Discordant findings may be explained by the lack of cardiac disease manifestation in aged mdx mice compared to WT, whereas significant cardiac dysfunction was previously seen with the sub-acute injury in young mice. Therefore, we are not able to conclude any cardioprotective effects with long-term nicorandil treatment in aging mdx mice.


Interactions of Phospholipid Vesicles with Cationic and Anionic Oligomeric Surfactants.
This work studied the interactions of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) with cationic ammonium surfactants and anionic sulfate or sulfonate surfactants of different oligomeric degrees, including cationic monomeric DTAB, dimeric C12C3C12Br2, and trimeric DDAD as well as anionic monomeric SDS, dimeric C12C3C12(SO3)2, and trimeric TED-(C10SO3Na)3. The partition coefficient P of these surfactants between the DOPC vesicles and water was determined with isothermal titration microcalorimetry (ITC) by titrating concentrated DOPC solution into the monomer solution of these surfactants. It was found that the P value increases with the increase of the surfactant oligomeric degree. Moreover, the enthalpy change and the Gibbs free energy for the transition of these surfactants from water into the DOPC bilayer become more negative with increasing the oligomeric degree. Meanwhile, the calcein release experiment proves that the surfactant with a higher oligomeric degree shows stronger ability of changing the permeability of the DOPC vesicles. Furthermore, the solubilization of the DOPC vesicles by these oligomeric surfactants was studied by ITC, turbidity, and dynamic light scattering, and thus the phase boundaries for the surfactant/lipid mixtures have been determined. The critical surfactant to lipid ratios for the onset and end of the solubilization for the DOPC vesicles derived from the phase boundaries decrease remarkably with increasing the oligomeric degree. Overall, the surfactant with a larger oligomerization degree shows stronger ability in incorporating into the lipid bilayer, altering the membrane permeability and solubilizing lipid vesicles, which provides comprehensive understanding about the effects of structure and shape of oligomeric surfactant molecules on lipid-surfactant interactions.


A C-H Insertion Approach to Functionalized Cyclopentenones.
Cyclopentenones are synthetically versatile structures, and their straightforward construction from alkynone substrates by employing synthetically streamlining C-H insertion is conceptually appealing and of high synthetic potential. But, its implementation is very limited. Herein we report a Au-catalyzed version, which affords 2-bromocyclopent-2-en-1-ones with a broad scope and synthetically desirable diastereoselectivities. The proposed key intermediate capable of the observed insertion into unactivated C-H bonds is a fully functionalized gold vinylidene, which is generated via a novel intermolecular strategy. This flexible access of likely gold vinylidenes opens various opportunities to explore their versatile reactivities.


Trendspotting.
All health care isn't local. To make the right strategic moves, you need to understand the bigger picture. A new environmental assessment from Deloitte & Touche and VHA, released first to Hospitals & Health Networks, sizes up the national scene.


Second-Order Disjoint Factor Analysis.
Hierarchical models are often considered to measure latent concepts defining nested sets of manifest variables. Therefore, by supposing a hierarchical relationship among manifest variables, the general latent concept can be represented by a tree structure where each internal node represents a specific order of abstraction for the latent concept measured. In this paper, we propose a new latent factor model called second-order disjoint factor analysis in order to model an unknown hierarchical structure of the manifest variables with two orders. This is a second-order factor analysis, which-respect to the second-order confirmatory factor analysis-is exploratory, nested and estimated simultaneously by maximum likelihood method. Each subset of manifest variables is modeled to be internally consistent and reliable, that is, manifest variables related to a factor measure "consistently" a unique theoretical construct. This feature implies that manifest variables are positively correlated with the related factor and, therefore, the associated factor loadings are constrained to be nonnegative. A cyclic block coordinate descent algorithm is proposed to maximize the likelihood. We present a simulation study that investigates the ability to get reliable factors. Furthermore, the new model is applied to identify the underlying factors of well-being showing the characteristics of the new methodology. A final discussion completes the paper.


Rejection in heart transplantation strongly correlates with HLA-DR antigen mismatch.
It is well established that incompatible HLA antigens presented by donor tissue readily evoke an immune response. Prospective HLA matching policies, widespread in European kidney transplant centers have reduced the level of HLA mismatching and have significantly improved graft survival. The influence of HLA incompatibility in heart transplantation remains controversial, and prospective HLA matching is seldom achieved. We examined the role of HLA antigen mismatching on transplant rejection by analyzing 2569 endomyocardial biopsies (EMB) from 157 consecutive orthotopic heart transplants performed from April 1987 to August 1993 in our own center. Biopsies were graded according to the accepted International Classification, with grade 2 and higher indicating rejection. Among 91 patients who received a 2 HLA-DR mismatch transplant 34% of 1624 biopsies analyzed were graded as > or = 2. This frequency fell to 29% of 797 biopsies for 53 patients with a one-HLA-DR mismatch and to 18% of 148 biopsies for 13 patients in the zero-HLA-DR-mismatch group (P < 0.00005). No significant effect on EMB grade frequencies was observed using the same method of analysis with transplants mismatched at the HLA-A or HLA-B loci apart from analysis of HLA-B matched transplants at 3 months posttransplant (P = 0.02). The close linkage of the HLA-B and HLA-DR loci may account for this observation. The results of this study show that heart transplants matched at the HLA-DR locus have a significantly reduced incidence of EMB grades indicative of rejection requiring augmented immunosuppressive therapy. We propose that prospective HLA-DR matching should be adopted for allocation of donor hearts for more efficient use of this precious and limited resource.


Unresponsiveness in hapten-specific cytotoxic T lymphocytes. I. Characteristics of tolerance induction in adult mice.
These experiments concern the regulation of cytotoxic T lymphocytes (CTL). Here we describe the induction of unresponsiveness in TNP-specific cytotoxic cells generated in vitro. Previous work has shown that i.v. injection of water-soluble TNP (TNBS) prevents in vivo generation of TNP-specific CTL in C3H/HeN mice. In contrast, this report shows that the in vitro generation of TNP-specific CTL from similarly treated C3H/HeN mice is not inhibited. Nevertheless, unresponsiveness in CTL can be induced with TNBS. This unresponsiveness appears to be H-2 linked, since 3 strains of H-2d mice (BALB/c, DBA/2J, and B10.D2) were rendered unresponsive after a single injection of TNBS, whereas 3 strains of H-2k mice (C58/J, C3H/HeN, and CBA/J) were not. This CTL unresponsiveness is antigen specific in both induction and expression. The unresponsiveness is reversed by pretreatment with cyclophosphamide but not by adult thymectomy. Furthermore, the addition of supernatants from Con A-stimulated spleen cells to cultures containing the unresponsive responder cells enhanced the generation of cytotoxic activity. These results suggest that a) the ability to induce TNP-specific CTL unresponsiveness is linked to H-2 loci, b) a regulatory network is involved in the unresponsiveness, which is sensitive to cyclophosphamide but resistant to adult thymectomy, and c) at least 1 level of regulation involves amplifying T cells.


Update on ISO 13485.
A previous article discussed the impact that new quality management system standards are likely to have on the CE-marking process. The draft international standard for medical device quality systems has now been published. This article will discuss important issues related to the new standard.


Characterization of the probabilistic traveling salesman problem.
We show that stochastic annealing can be successfully applied to gain new results on the probabilistic traveling salesman problem. The probabilistic "traveling salesman" must decide on an a priori order in which to visit n cities (randomly distributed over a unit square) before learning that some cities can be omitted. We find the optimized average length of the pruned tour follows E(L(pruned))=sqrt[np](0.872-0.105p)f(np), where p is the probability of a city needing to be visited, and f(np)-->1 as np--> infinity. The average length of the a priori tour (before omitting any cities) is found to follow E(L(a priori))=sqrt[n/p]beta(p), where beta(p)=1/[1.25-0.82 ln(p)] is measured for 0.05< or =p< or =0.6. Scaling arguments and indirect measurements suggest that beta(p) tends towards a constant for p<0.03. Our stochastic annealing algorithm is based on limited sampling of the pruned tour lengths, exploiting the sampling error to provide the analog of thermal fluctuations in simulated (thermal) annealing. The method has general application to the optimization of functions whose cost to evaluate rises with the precision required.


Genetic Expression Screening of Arsenic Trioxide-Induced Cytotoxicity in KG-1a Cells Based on Bioinformatics Technology.
Acute myeloid leukemia (AML) is a malignant tumor of the hematopoietic system, and leukemia stem cells are responsible for AML chemoresistance and relapse. KG-1a cell is considered a leukemia stem cell-enriched cell line, which is resistant to chemotherapy. Arsenic trioxide (ATO) is effective against acute promyelocytic leukemia as a first-line treatment agent, even as remission induction of relapsed cases. ATO has a cytotoxic effect on KG-1a cells, but the mechanism remains unclear. Our results demonstrated that ATO can inhibit cell proliferation, induce apoptosis, and arrest KG-1a cells in the G2/M phase. Using transcriptome analysis, we investigated the candidate target genes regulated by ATO in KG-1a cells. The expression profile analysis showed that the ATO had significantly changed gene expression related to proliferation, apoptosis, and cell cycle. Moreover, MYC, PCNA, and MCM7 were identified as crucial hub genes through protein-protein interaction network analysis; meanwhile, the expressions of them in both RNA and protein levels are down-regulated as confirmed by quantitative polymerase chain reaction and Western blot. Thus, our study suggests that ATO not only inhibits the expression of MYC, PCNA, and MCM7 but also leads to cell cycle arrest and apoptosis in KG-1a cells. Overall, this study provided reliable clues for improving the ATO efficacy in AML.


Malakoplakia of the prostate on needle core biopsy: a case report and review of the literature.
Malakoplakia is a rare inflammatory disorder that results from an infectious process and that may be associated with immunosuppression. It most often occurs in the genitourinary tract; however, isolated cases have been reported in many other organs, including colon, stomach, lung, liver, bone, uterus, and skin. In the bladder, which is the most common site of involvement, it may be mistaken for malignancy on cystoscopic examination or computerized topography scan. Gram-negative bacteria such as Escherichia coli and Klebsiella pneumonia are often isolated from malakoplakia lesions, although other less common bacteria such as Rhodococcus equi may also be found. This is a case report of prostatic malakoplakia diagnosed on needle core biopsy from a patient suspected to harbor a malignancy.


'Cultural safety' and the analysis of health policy affecting aboriginal people.
Vicki Smye and Annette Browne explore the exportability of the concept of 'cultural safety' from the healthcare literature in New Zealand to inform an analysis of mental health policy discourse affecting aboriginal communities in British Columbia, Canada. The moral issues raised are, they suggest, ones that nurses in research, policy and practice must attend to when providing health care to marginalised, disenfranchised populations.