Pulsed multifrequency thermosonication induced sonoporation in Alicyclobacillus acidoterrestris spores and vegetative cells.
Alicyclobacillus acidoterrestris (AAT) was proposed as an index of pasteurization design for high-acid fruit products due to its spore resistance and repeated spoilage incidences in fruit juices. This study aimed to determine the effectiveness of pulsed multifrequency ultrasound to minimize AAT spores and vegetative cells in aqueous suspension. For this research, an investigation of the reactive oxygen species and antioxidant activity was performed to examine the effect of temperature and frequency on AAT spore inoculation. Total decreases in AAT bacteria were 5.99, 5.74 Log CFU/mL in vegetative cells for dual-frequency thermosonication (DFTS) and dual-frequency ultrasonication (DFUS), respectively, while 5.90 and 5.38 Log CFU/mL in spores for both DFUS and DFTS, respectively. The loss of the percentage of cells in ultrasound (US) and thermosonication (TS) treatments was inversely associated with the rate of O2-and H2O2 development. The fluorescence microscopy revealed a higher bactericidal efficacy of DFTS compared to the DFUS and control. Scanning Electron Microscopy (SEM) and Transmission electron microscopy (TEM) demonstrated ultra-structural modifications such as the interruption of cell walls by cavitation and pores in the membrane structure of the AAT bacteria induced by sonoporation. Several TS frequencies of 20/40/60, 20/40, and 20 kHz treated spores had a higher electrical conductivity than untreated ones, with an improvement of 7.94, 5.68, and 3.72 %, respectively. Fourier-transform infrared (FTIR) spectroscopy revealed major changes in the spectral region of membrane fatty acids and proteins of AAT. Simultaneously, AAT inactivation specific energy rate was significantly reduced using dual-frequency ultrasound, compared to mono-frequency thermosonication. The significant results of this work recommended pulsed DFUS as an alternative application to mono-frequency US in beverage industries.


Epidemiological characteristics of respiratory viruses in hospitalized children during the COVID-19 pandemic in southwestern China.
Multinational studies have reported that the implementation of nonpharmaceutical interventions (NPIs) to control severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission coincided with the decline of other respiratory viruses, such as influenza viruses and respiratory syncytial virus. To investigate the prevalence of common respiratory viruses during the coronavirus disease 2019 (COVID-19) pandemic. Respiratory specimens of children with lower respiratory tract infections (LRTIs) hospitalized at the Children's Hospital of Chongqing Medical University from January 1, 2018 to December 31, 2021 were collected. Seven common pathogens, including respiratory syncytial virus (RSV), adenovirus (ADV), influenza virus A and B (Flu A, Flu B), and parainfluenza virus types 1-3 (PIV1-3), were detected by a multiplex direct immunofluorescence assay (DFA). Demographic data and laboratory test results were analyzed. 1) A total of 31,113 children with LRTIs were enrolled, including 8141 in 2018, 8681 in 2019, 6252 in 2020, and 8059 in 2021.The overall detection rates decreased in 2020 and 2021 (P < 0.001). The detection rates of RSV, ADV, Flu A, PIV-1, and PIV-3 decreased when NPIs were active from February to August 2020, with Flu A decreasing most predominantly, from 2.7% to 0.3% (P < 0.05). The detection rates of RSV and PIV-1 resurged and even surpassed the historical level of 2018-2019, while Flu A continued decreasing when NPIs were lifted (P < 0.05). 2) Seasonal patterns of Flu A completely disappeared in 2020 and 2021. The Flu B epidemic was observed until October 2021 after a long period of low detection in 2020. RSV decreased sharply after January 2020 and stayed in a nearly dormant state during the next seven months. Nevertheless, the detection rates of RSV were abnormally higher than 10% in the summer of 2021. PIV-3 decreased significantly after the COVID-19 pandemic; however, it atypically surged from August to November 2020. The NPIs implemented during the COVID-19 pandemic affected the prevalence and seasonal patterns of certain viruses such as RSV, PIV-3, and influenza viruses. We recommend continuous surveillance of the epidemiological and evolutionary dynamics of multiple respiratory pathogens, especially when NPIs are no longer necessary.


The binding of calcium ions to bovine factor X by rate dialysis.
The binding of Ca+2 to bovine factor X (molecular weight of 74,000) (Yue und Gertler 1977) was studied by the technique of rate dialysis and with the use of 45Ca+2. The binding data are consistent with a model of sequential mechanism. One mole of Ca+2 binds to the glycoprotein with a dissociation constant of 5.2 X 10(-5) M and additional 39 +/- 4 moles of Ca+2 bind to this zymogen with a dissociation constant of 3.7 X 10(-3) M. The binding of the high affinity Ca+2 causes a functionally significant change in the zymogen, and (calcium) (factor X) complex is the real substrate in the activation process by the protease in Russell's viper venom.


Application of a solid-phase radioimmunoassay and immune electron microscopy for hepatitis A in diagnosis and research.
With crude virus suspensions from stool and antibodies from hepatitis-A patients, a solid-phase radioimmunoassay (RIA) for detection of hepatitis virus A (HVA) had antibodies against hepatitis virus A (anti-HVA) has been developed. Examples for the application of this test are demonstrated. Virus particles from the stools of the two patients were further characterized. Serologically, they were identical or very similar to the MS-1 strain. Isopycnic CsCl-gradient centrifugation of both strains revealed two peaks, but the particles of different densities did not differ in size or serologically. A modification of the RIA was also useful for determination of IgM antibodies in patients' sera fractionated by sucrose-density centrifugation. The application of the RIA method for serologic epidemiology is demonstrated by a comparison of anti-HVA prevalence in German and non-German women residing in Germany.


Family physician continuity of care and emergency department use in end-of-life cancer care.
Despite cancer patients preferring to spend their last days out-of-hospital, many make difficult visits to the emergency department (ED). Family physician continuity of care has been shown in some clinical situations to reduce ED utilization. To determine if greater family physician continuity of care for cancer patients during the end-of-life is associated with less ED utilization. This retrospective, population-based study involved secondary analysis of linked administrative data files for 1992 to 1997. Sources included the Nova Scotia Cancer Registry, Vital Statistics, the Queen Elizabeth II Health Sciences Center Oncology Patient Information System and Palliative Care Program (PCP), Hospital Admissions/Separation data, and Physician Services information. Subjects included adults with a recorded date of cancer diagnosis who died of cancer and who had made at least three visits to a family physician during their last 6 months of life. The relationship between total ED visits and family physician continuity of care, developed using the Modified Modified Continuity Index (MMCI), was examined using negative binomial regression with adjustments for survival, year of death, sex, age, cancer type, region, PCP admission, specialty visits, hospital days, death location, income quintile, and total ambulatory visits. In total, 8702 subjects made 11,551 ED visits (median = 1.0); median MMCI was 0.83. Adjusted results indicate those experiencing low continuity (MMCI < 0.5) made 3.9 times more ED visits (rate ratio [RR] = 3.93; 95% CI [CI] = 3.57-4.34) than those experiencing high continuity (MMCI > or = 0.8) and patients experiencing moderate continuity (MMCI = 0.5-0.8) made twice as many ED visits (RR = 2.28; CI = 2.15-2.42). Given this significant association between family physician continuity of care and ED visits during the end-of-life, and given international trends to reform primary care, active planning of strategies to facilitate such continuity should be encouraged.


[Preparation of alpha-hydrocycholic-beta-cyclodextrin inclusion compound].
To study the preparation of alpha-Hydrocycholic-beta-Cyclodextrin inclusion compound. It was studied with orthogonal design to analysis three factors of inclusion rate such as the weight ratio between HDCA and beta-Cyclodextrin, the temperature and the reaction time. Fourier transform infrared spectroscopy, scanning electron microscopy, taste and solubility test were used to identify the inclusion compound. Stable inclusion compound was made by HDCA and beta-Cyclodextrin. The weight ratio between HDCA and beta-Cyclodextrin was the most important factor. The alpha-Hydrocycholic-beta-Cyclodextrin inclusion compound taste and the solubility were improved. The optimum preparation condition is alpha-Hydrocycholic : beta-Cyclodextrin = 8.67 : 1, the inclusion temperature is 60 degrees C and the inclusion time is 30 minutes.


Quantitative determination of 5-aminolaevulinic acid and its esters in cell lysates by HPLC-fluorescence.
The development of a reliable sensitive method for the HPLC determination of 5-aminolaevulinic acid (ALA) and ALA esters in cell lysates is described. The method relies on the quantification of a fluorescent derivative of ALA following its derivatisation with acetylacetone and formaldehyde. Following this procedure it is possible to quantify ALA in cell lysates with no need for pre-purification of the sample. The method has been validated in two ranges of concentration (0.6-65 microM, 0.1-10 microg/mL, and 30-600 microM, 5-100 microg/mL), and has also been extended and validated for the determination of ALA released from ALA prodrugs after acidic hydrolysis.


Cannulated percutaneous fixation of intra-articular hand fractures.
Percutaneous fixation techniques minimize edema, scar formation, and stiffness from operative trauma when restoring position and stability of displaced and unstable hand fractures. Percutaneous Kirschner wires merely splint fractures and may migrate, disengage, or irritate soft tissues; may be associated with pin tract suppuration; and may require removal at 4 to 6 weeks after insertion. Percutaneous miniscrews may provide more secure fixation for larger articular and oblique diaphyseal fractures of the thumb and finger phalanges by means of bicortical fixation and compression, are less intrusive to adjacent tissues, retain the fracture throughout the healing process, and seldom require removal. Miniscrew canulation substantially simplifies the insertion process, and headless miniscrews are entirely unobtrusive to the adjacent tissues.


Sustained liver-specific transgene expression from the albumin promoter in mice following hydrodynamic plasmid DNA delivery.
To properly study gene expression in vivo, often long-term expression is desired. Previous studies using plasmid DNA (pDNA) vectors have typically resulted in short-term expression. Here, we evaluated combinations of the albumin promoter with different enhancers and untranslated regions for liver-specific expression in mice. A series of pDNA secreted alkaline phosphatase (SEAP) reporter gene expression vectors was constructed using the albumin promoter and various other expression cassette elements. Each was evaluated for level and duration of SEAP expression in mice following hydrodynamic tail vein delivery. Sustained liver expression was obtained from vectors combining the albumin promoter with an albumin 3' untranslated region (3'UTR). The level of expression was increased by inclusion of enhancers and a 5' intron. The optimal expression vector consisted of the albumin promoter combined with an alpha-fetoprotein MERII enhancer, 5' intron from the factor IX gene, and the 3'UTR from the albumin gene including intron 14. With this vector, SEAP reporter gene expression levels remained high for 1 year, at levels comparable to those obtained from the cytomegalovirus (CMV) promoter on day 1. Expression of human apolipoprotein E3 (hApoE) in ApoE knockout mice provided a dose-dependent correction of their hypercholesterolemia. Liver-specific sustained transgene expression can be obtained at very high levels from optimized pDNA vectors, without the use of integration systems. Such vectors will further facilitate biological studies of genes in vivo and may find application in gene therapy.


Mutagenicity, carcinogenicity and reproductive effects of dibromochloropropane (DBCP).
Dibromochloropropane (DBCP), a brominated organochlorine nematocide, has been used since the mid-1950s. Its primary value was its effectiveness on perennial crops without damaging the plants. DBCP also had less acute toxicity than earlier soil fumigants such as ethylene dibromide. DBCP became widely used on citrus, grapes, peaches, pineapple, soybeans and tomatoes. In Central America and Israel, banana trees were treated with DBCP. By 1975, 25 million pounds were being produced in the U.S. per year. The majority of U.S. production was applied in the Pacific Coast states, particularly California's San Joaquin Valley, and the Southern Atlantic Coast states. Animal studies in the early 1960s revealed hepatic and renal effects and testicular atrophy in DBCP-treated rates. Testicular atrophy was noted at the lowest exposure level tested (5 ppm). In 1973, DBCP was found to cause gastric carcinoma in mice and rats, and mammary adenocarcinoma in rats. A later study linked DBCP to nasal cavity tumors in rats. DBCP is an in vitro mutagen after activation with microsomal enzymes. A dominant lethal effect has been observed in rats. DBCP also induces increases in sister-chromatid exchanges and chromosomal aberrations. Mean fetal weight and maternal weight gain is decreased in pregnant rats exposed to DBCP. In mid-1977, employees in a California pesticide formulation plant were discovered to be infertile. Further investigations documented azoospermia and oligospermia among many of the workers handling DBCP. These findings were duplicated in other DBCP production plants. In the United States, the Occupational Safety and Health Administration (OSHA) and the Environmental Protection Agency (EPA) rapidly restricted the use of DBCP. The application of DBCP to all but pineapples in Hawaii was banned by the EPA in 1979. In 1982, the EPA exempted the peach crop in the Southern Atlantic coast states from the ban on DBCP use. Prior to the 1977 events, no such agent in workplace concentrations had been shown to produce infertility and sterility in otherwise healthy men. Subsequently, the interest in DBCP had led to many other studies, and has contributed to a heightened awareness about workplace hazards to both the male and female reproductive systems. This article will review the current state of knowledge of the in vitro, animal, and human toxicology of DBCP.


Comparative columellar scar analysis between transverse and inverted-V incision in open rhinoplasty.
Rhinoplasty can be performed using both open and closed approaches. A visible scar on the columella is the major disadvantage of open rhinoplasty. Different columellar incision types have been used in open rhinoplasty. In this retrospective study, we compare transverse and inverted-V columellar incisions with a scar assessment scale. In this retrospective study, open rhinoplasty was performed on 84 patients between 2001 and 2006. The transverse incision was used on 39 patients (18 males, 21 females). The inverted-V incision was used on 45 patients (21 males, 24 females). The entire surgical procedure was performed by a single surgeon. The columellar incision was closed using 6-0 interrupted polypropylene sutures. All sutures were removed on the fifth postoperative day. With this assessment scale, we observed satisfactory scar, pigmentation, and notching. We compared both groups and found that the inverted-V incision resulted in better scar formation (p < 0.05) and less notching (p < 0.07). Scar pigmentation was found to be irrelevant to the incision technique employed (p < 0.3). As a result of this study we concluded that the inverted-V incision might be a better choice in open rhinoplasty.


Giardia lamblia.
Giardia is the most frequently identified enteric parasite in the United States, but much is not known about host-parasite interaction. Advances using immunodiagnostic techniques, endonuclease restriction analysis, surface-antigen detection methods, and measuring homologous and heterologous antibody responses are beginning to clarify the biology of Giardia.


Social cohesion, mental wellbeing and health-related quality of life among a cohort of social housing residents in Cornwall: a cross sectional study.
Research and policy have identified social cohesion as a potentially modifiable determinant of health and wellbeing that could contribute to more sustainable development. However, the function of social cohesion appears to vary between communities. The aim of this study was to analyse the levels of, and associations, between social cohesion, mental wellbeing, and physical and mental health-related quality of life among a cohort of social housing residents from low socioeconomic status communities in Cornwall, UK. Social housing is below market-rate rental accommodation made available to those in certain health or economic circumstances. These circumstances may impact on the form and function of social cohesion. During recruitment, participants in the Smartline project completed the Short Warwick-Edinburgh Mental Wellbeing Scale, SF-12v2 and an eight item social cohesion scale. Cross sectional regression analyses of these data adjusted for gender, age, national identity, area socioeconomic status, rurality, education, employment, and household size were undertaken to address the study aim. Complete data were available from 305 (92.7%) participants in the Smartline project. Univariable analyses identified a significant association between social cohesion, mental wellbeing and mental health-related quality of life. Within fully adjusted multivariable models, social cohesion only remained significantly associated with mental wellbeing. Sensitivity analyses additionally adjusting for ethnicity and duration of residence, where there was greater missing data, did not alter the findings. Among a relatively homogeneous cohort, the reported level of social cohesion was only found to be significantly associated with higher mental wellbeing, not physical or mental health-related quality of life. The efforts made by social housing providers to offer social opportunities to all their residents regardless of individual physical or mental health state may support the development of a certain degree of social cohesion. Sense of control or safety in communities may be more critical to health than social cohesion. Additional observational research is needed before attempts are made to alter social cohesion to improve health.


A 58-Year-Old Man With Hemoptysis After COVID-19 Infection.
A 58-year-old man presented to us with a 1-week history of high-grade fever and progressive dry cough. Four weeks before his presentation, he was diagnosed with COVID-19 infection and needed non-ICU hospital admission with no supplemental oxygen requirements for 6 days and was treated with a 5-day course of remdesivir and 3 weeks of dexamethasone. His steroid dose was commenced on dexamethasone 12 mg bid (four times the recommended dose) for 14 days and then gradually tapered over the remaining 7 days. His history was unremarkable, except for well-controlled asthma. He did not complain of any shortness of breath, weight loss, or loss of appetite. He was never a smoker and denied any alcohol use.


Prognostic biomarkers for malignant transformation of oral potentially malignant disorders: a scoping review protocol.
This scoping review aims to identify and systematically map the available evidence concerning the prognostic biomarkers for malignant transformation of oral potentially malignant disorders (OPMDs), and to identify and analyze possible knowledge gaps in this field of literature. It is hypothesized that diagnosis and treatment of oral cancer in its early stages may be the key to improving the prognosis and reducing treatment-related consequences. Oral potentially malignant disorders represent tissue alterations with a higher risk of malignant transformation compared to the normal mucosa. Therefore, the study of prognostic biomarkers for OPMD could represent new diagnosis and therapeutic targets and, consequently, contribute to the reduction of oral cancer burden worldwide. Longitudinal studies investigating prognostic biomarkers regarding the malignant transformation of OPMD will be included. The initial OPMD diagnosis and the malignant transformation must have been confirmed by histopathological analysis. To achieve minimal heterogeneity, studies that assess biomarkers in other locations (blood, plasma or others) will be excluded. Five electronic databases and three grey literature databases will be consulted. No restrictions regarding publication date will be applied. Only studies published in the Latin (Roman) alphabet, which comprises most of the European languages, will be included. Study selection will be performed by two authors in a two-phase process; if any disagreement arises, a third author will be consulted to make a final decision. Data extraction will be performed by two authors using a standardized extraction tool. The results will be described in details accordantly with the aims of this scoping review.


Non-governmental organizations in the health field: collaboration, integration and contrasting aims.
Non-governmental organizations, from being voluntary agencies often with relief or religious mission orientations, have become very important players in the field of social development. In this paper something is given of their evolution, diversity, aims and activities; and of the important role they now play in many instances, as well as some of the obstacles to collaboration, co-ordination and integration at different levels of operation--internationally, nationally and locally.


Stability-Indicating Analytical Method Development Using Quality by Design (QbD) Approach for Simultaneous Estimation of Budesonide and Levosalbutamol.
Budesonide is a corticosteroid; levosalbutamol is bronchodilator. In combination, they are used in the treatment of asthma and chronic obstructive pulmonary disease. The present article portrays the development of a stability-indicating high-performance thin-layer chromatography (HPTLC) method for the simultaneous estimation of budesonide and levosalbutamol using design of experiment (DOE) principles. DOE was applied in development part in which a Box-Behnken design was used to study the effect of factors on the Rf value of the drugs. Twenty-five experimental runs were performed to optimize the chromatographic conditions. Aluminum sheets precoated with silica gel 60 F254 were used as the stationary phase. The optimized mobile phase composition was found to be toluene-ethyl acetate-methanol-ammonia (4:1.8:1.8:0.2) and saturation time 20 min, quantified by densitometric analysis at 231 nm. Moreover, the drugs were subjected to acid and alkali hydrolysis, oxidation, thermal, and photodegradation. The drugs undergo degradation under mainly acidic and basic conditions. Also, the degraded products were well resolved from the pure drug with significantly different Rf values. Linearity was performed in the ranges of 224-1120 ng/band for budesonide and 280-1400 ng/band for levosalbutamol. The method was validated according to the International Conference on Harmonization guidelines. This approach is useful to expedite method development and optimization activities in analytical laboratories. The experimental data revealed that the volume of toluene and methanol in the mobile phase significantly affected the Rf value of both the drugs, and ammonia significantly affected the Rf value of levosalbutamol.


Melanoma cell necrosis facilitates transfer of specific sets of antigens onto MHC class II molecules of dendritic cells.
Vaccine strategies that target dendritic cells (DC) in order to elicit immunity against tumors are the subject of intense research. For the induction and maintenance of anti-tumor immunity, CD4+ helper T cells are often required, which need to see appropriate MHC class II-peptide complexes on DC. So far, it remained widely unclear what type of tumor cells can feed the MHC class II processing pathway of DC with what type of antigens. Here, we report that peptide loading onto MHC class II molecules of myeloid DC is facilitated by melanoma cells undergoing necrotic rather than apoptotic cell death. Importantly, the set of MHC class II-associated peptides induced by necrotic tumor cells differed from those found upon engagement of apoptotic tumor cells. This may be due to the fact that only necrotic cells liberated heat shock proteins, which bind tumor-derived peptides and thereby may promote processing by DC. The potential of DC to activate T cells was kinetically controlled through their antigen receptivity: CD4+ T cells were easily stimulated upon encountering antigen early in DC maturation, whereas antigen capture at later maturation stages favored activation of CD8+ T cells. These findings may aid in designing future vaccination strategies and in identifying novel tumor-specific helper T cell antigens.


Viral gastroenteritis--a danger to the patient, a danger to the staff.
Viral gastroenteritis is usually a mild, self-limiting illness. We report a case of a previously well 74-year-old woman who suffered a grave complication of this common condition, and caused a significant outbreak of illness amongst staff involved in her care. This case highlights the risks of Hospital Acquired Infection and raises important infection control issues. It illustrates the hazards associated with exposure to potentially infectious secretions and presents a clear message to medical and nursing staff involved in the care of the acute surgical emergency.


Nanohybrids Composed of Tannic Acid Cross-Linked by Metal Ions Obtained by a Microfluidic Technique.
Naturally occurring compounds, such as tannic acid (TA), are perfect for constructing nanohybrids (NHs) with metal ions due to their anticarcinogenic, antimicrobial, and antioxidant properties. To date, the batch methods are the ones in which such NHs were constructed; however, those methods possess many drawbacks, such as low reproducibility or size variations. To overcome this limitation, microfluidic preparation is proposed for NHs construction composed of TA and iron (III). The spherical particles with a size between 70 and 150 nm and antimicrobial properties can be easily fabricated in a controlled manner.


Ifosfamide alone and in combination in the treatment of refractory malignant gestational trophoblastic disease.
We attempted to evaluate the use of ifosfamide either alone or in combination in patients with refractory malignant gestational trophoblastic disease. Our study comprised, in part, a phase II multiinstitutional trial of ifosfamide in refractory gynecologic malignancies and, in part, a review of institutional experience with ifosfamide in combination chemotherapy. Single-agent ifosfamide produced a significant response in titer in one of two patients with refractory choriocarcinoma. Ifosfamide with etoposide and cisplatin (also known as VIP) resulted in significant response in human chorionic gonadotropin titers in three patients with highly refractory metastatic gestational trophoblastic disease and one cure in this group of patients. Ifosfamide has activity in refractory choriocarcinoma and, when combined with etoposide and cisplatin (VIP), may be curative.


Strategies of fluorescence staining for trace total ribonucleic acid analysis by capillary electrophoresis with argon ion laser-induced fluorescence.
In this work, five fluorescent dyes (SYTO-9, SYBR Green I, SYBR Green II, SYBR Safe, and SYBR Gold) were used as both on-column and precolumn stains for total RNA analysis by CE-LIF with Ar ion laser excitation. In the on-column RNA stain, the SYTO-9 provided the highest fluorescence intensity and the lowest detectable concentration, as low as 10 pg/μL, while the SYBR Green II and SYBR Gold were adsorbed on the poly(ethylene oxide) thus affected the separation efficiency. As a precolumn stain, SYBR Gold was the most sensitive among the five dyes due to the strong affinity between the dye and RNA molecules. As a result, a single-cell quantity of RNA (10-30 pg per cell) could be detected by CE-LIF with precolumn staining by SYBR Gold. Because of the great savings of fluorescent dye using precolumn stain (one button dye may use for one million stain), this method is the best strategy for RNA staining in terms of cost-effectiveness and sensitivity.


Mitochondrial genome polymorphisms associated with type-2 diabetes or obesity.
Although a strong correlation between type-2 diabetes and obesity has been found, no comparative analysis between diabetes and obesity has been performed with respect to mitochondrial DNA (mtDNA) polymorphisms. To test the hypothesis that certain mitochondrial single nucleotide polymorphisms (mtSNPs) might be associated with obesity or type-2 diabetes, we determined the entire sequences of the mitochondrial genomes from 96 patients with type-2 diabetes and those from 96 young obese adults by direct sequencing and compared the frequencies of mtSNPs between these two groups. A mtSNP, 8684C > T (T53I) in the mitochondrial ATP synthase subunit 6 gene (ATP6), was detected in 5 of the 96 patients with type-2 diabetes, whereas this substitution was not detected in any of the 96 young obese adults. Two mtSNPs, 3497C > T (A64V) in NADH dehydrogenase subunit 1 gene (ND1) and 1119T > C (472U > C) in the 12S rRNA gene, were detected in 5 of the 96 young obese adults, whereas these substitutions were not detected in any of the 96 diabetic patients. The 8684C > T transition associated with type-2 diabetes represents haplogroup M8a, and the 3497C > T and 1119T > C transitions predisposing to obesity represent haplogroup B4c. These results suggest that distinct mtSNPs contribute to susceptibility to type-2 diabetes or obesity, pointing out the necessity of large-scale case control studies.


Characterization of a novel inhibitor for the New Delhi metallo-β-lactamase-4: Implications for drug design and combating bacterial drug resistance.
The bacterial metallo-β-lactamases (MBLs) catalyze the inactivation of β-lactam antibiotics. Identifying novel pharmacophores remains crucial for the clinical development of additional MBL inhibitors. Previously, 1-hydroxypyridine-2(1H)-thione-6-carboxylic acid, hereafter referred to as 1,2-HPT-6-COOH, was reported as a low cytotoxic nanomolar β-lactamase inhibitor of Verona-integron-encoded metallo-β-lactamase 2, capable of rescuing β-lactam antibiotic activity. In this study, we explore its exact mechanism of inhibition and the extent of its activity through structural characterization of its binding to New Delhi metallo-β-lactamase 4 (NDM-4) and its inhibitory activity against both NDM-1 and NDM-4. Of all the structure-validated MBL inhibitors available, 1,2-HPT-6-COOH is the first discovered compound capable of forming an octahedral coordination sphere with Zn2 of the binuclear metal center. This unexpected mechanism of action provides important insight for the further optimization of 1,2-HPT-6-COOH and the identification of additional pharmacophores for MBL inhibition.


Integrate life-cycle assessment and risk analysis results, not methods.
Two analytic perspectives on environmental assessment dominate environmental policy and decision-making: risk analysis (RA) and life-cycle assessment (LCA). RA focuses on management of a toxicological hazard in a specific exposure scenario, while LCA seeks a holistic estimation of impacts of thousands of substances across multiple media, including non-toxicological and non-chemically deleterious effects. While recommendations to integrate the two approaches have remained a consistent feature of environmental scholarship for at least 15 years, the current perception is that progress is slow largely because of practical obstacles, such as a lack of data, rather than insurmountable theoretical difficulties. Nonetheless, the emergence of nanotechnology presents a serious challenge to both perspectives. Because the pace of nanomaterial innovation far outstrips acquisition of environmentally relevant data, it is now clear that a further integration of RA and LCA based on dataset completion will remain futile. In fact, the two approaches are suited for different purposes and answer different questions. A more pragmatic approach to providing better guidance to decision-makers is to apply the two methods in parallel, integrating only after obtaining separate results.


Interleukin-2, anti-interleukin-2 receptor antibody, and activation of macrophages.
Macrophages treated with IFN-gamma and IL-2 before exposure to parasites develop the ability to resist infection with amastigotes of Leishmania major. In this cooperative interaction of cytokines, IL-2 can be replaced with any of several mAb directed against the beta chain of the IL-2 receptor, but not by antibodies to a number of other cell receptors or antigens. Thus, antibodies to the IL-2 receptor act as agonists of IL-2 in the induction of a biologic activity in macrophages, and macrophages, a nonproliferating cell type, respond to signals transmitted through the beta chain of the IL-2 receptor.


Weighing in on ubiquitin: the expanding role of mass-spectrometry-based proteomics.
Mass-spectrometry-based proteomics has become an essential tool for the qualitative and quantitative analysis of cellular systems. The biochemical complexity and functional diversity of the ubiquitin system are well suited to proteomic studies. This review summarizes advances involving the identification of ubiquitinated proteins, the elucidation of ubiquitin-modification sites and the determination of polyubiquitin chain linkages, as well as offering a perspective on the application of emerging technologies for mechanistic and functional studies of protein ubiquitination.


Desiccation survival of the infective larvae of the insect parasitic nematode, Heterorhabditis zealandica poinar.
Desiccation survival of the insect parasitic nematode Heterorhabditis zealandica upon exposure to various relative humidities on membrane filters and agar plates was determined. Survival was poor once water had been lost from the substrate. A screening experiment indicates that survival would not be improved by adjusting the treatment temperature, the source of the infective larvae, the method of rehydration or the addition of trehalose. Desiccation does not, therefore, appear to be a promising option for the long-term storage of these nematodes.


RBFOX2 recognizes N6-methyladenosine to suppress transcription and block myeloid leukaemia differentiation.
N6-methyladenosine (m6A) methylation can be deposited on chromatin-associated RNAs (caRNAs) by the RNA methyltransferase complex (MTC) to regulate chromatin state and transcription. However, the mechanism by which MTC is recruited to distinct genomic loci remains elusive. Here we identify RBFOX2, a well-studied RNA-binding protein, as a chromatin factor that preferentially recognizes m6A on caRNAs. RBFOX2 can recruit RBM15, an MTC component, to facilitate methylation of promoter-associated RNAs. RBM15 also physically interacts with YTHDC1 and recruits polycomb repressive complex 2 (PRC2) to the RBFOX2-bound loci for chromatin silencing and transcription suppression. Furthermore, we found that this RBFOX2/m6A/RBM15/YTHDC1/PRC2 axis plays a critical role in myeloid leukaemia. Downregulation of RBFOX2 notably inhibits survival/proliferation of acute myeloid leukaemia cells and promotes their myeloid differentiation. RBFOX2 is also required for self-renewal of leukaemia stem/initiation cells and acute myeloid leukaemia maintenance. Our study presents a pathway of m6A MTC recruitment and m6A deposition on caRNAs, resulting in locus-selective chromatin regulation, which has potential therapeutic implications in leukaemia.


Characteristics of users of a tailored, interactive website for parents and its impact on adolescent vaccination attitudes and uptake.
We examined the characteristics of parents using an iPad-based intervention about vaccines, and its impact on vaccination attitudes and behavior. Interventions were implemented in three primary care clinics from June 2012-September 2013. Baseline and follow up surveys assessed vaccination attitudes and intentions. Medical records were used to examine adolescent vaccine uptake. During the study, 42 parents viewed tailored educational content. Users were generally positive about vaccines, though one out of five worried that vaccines caused more harm than good. Among the 16 parents completing the post-intervention survey, there was a slightly higher, non-statistically significant, mean vaccination intention after viewing the website than prior to viewing it for three of the four adolescent vaccines (all except tetanus-diphtheria-acellular pertussis). Using the intervention did not increase the likelihood of adolescent vaccination. Providing educational material via iPads in clinic waiting rooms does not appear to be an effective strategy for engaging parents about vaccines. Overall, parents' interaction with TeenVaxScene was low, and had little impact on their vaccination attitudes and beliefs. However, use of TeenVaxScene did not appear to worsen parents' attitudes about vaccines. New and creative ideas for engaging parents to use such educational materials are needed.


Optimal conditions for determination of zinc bacitracin, polymyxin B, oxytetracycline and sulfacetamide in animal feed by micellar electrokinetic capillary chromatography.
A separation technique for zinc bacitracin, polymyxin B, oxytetracycline and sulfacetamide in animal feedstuffs by micellar electrokinetic capillary chromatography (MEKC) was developed. The running buffer was 20 mmol l(-1) borate, 20 mmol l(-1) phosphate, pH 8.4, containing 20 mmol l(-1) sodium dodecylsulphate and 10% (v/v) methanol. MEKC was performed at 25 degrees C; the applied voltage was 25 kV with a running pressure of 10 mbar. Simultaneous UV detection for all analytes was at 215 nm. The method was validated for specificity, accuracy, linearity, precision and robustness. It was shown to be specific, accurate (recoveries were 99.7 +/- 0.3, 99.9 +/- 0.9, 99.8 +/- 1.0 and 99.5 +/- 0.4, respectively, for oxytetracycline-, sulfacetamide-, polymyxin B- and zinc bacitracin-spiked samples of feed for cow, pigs, chicken and cattle), linear over the tested range (correlation coefficients > or =0.9987) and precise (RSDs below 1.8% for each analyte). The method was applied to determine zinc bacitracin, polymyxin B, oxytetracycline and sulfacetamide as additives in animal feed.


Does previous circumcision and wealth index influence women's attitude to discontinue the practice of female genital mutilation and cutting (FGM/C) in Ethiopia?
understanding women's attitudes towards female genital mutilation is an important step towards eliminating this practice. We used the 2016 Ethiopia Demographic and Health Survey (EDHS) data set to examine the relationship between wealth index, and previous history of circumcision on women's opinions whether female genital mutilation (FGM) should be continued or stopped in Ethiopia. Data from 6984 women aged 15-49 years were extracted from the 2016 Ethiopia EDHS data set. Multivariable logistic regression analysis was performed to analyse the data. In this study, women with a higher level of education and wealth index were more likely to support the cessation of FGM. However, circumcised women (AOR: 0.22; 95% CI: 0.15-0.32), women from the Afar region (AOR: 0.34; 95% CI: 0.22-0.50), Somali region (AOR: 0.42; 95% CI: 0.27-0.65), and Dire Dawa region (AOR: 0.51; 95% CI: 0.32-0.83) were less likely to support discontinuation of FGM. The present study revealed that wealth index, education level, history of circumcision, and regional variation are associated with women's attitude towards discontinuation of the practice of FGM in Ethiopia. Empowering women in terms of socioeconomic status and education can change attitudes and might help prevent female genital mutilation in the future. Furthermore, interventions targeting FGM practices should focus on regional variance in order to have a meaningful impact on reducing this harmful cultural practice in Ethiopia.


Topographical electrogastrograms after radical esophagectomy with colonic replacement.
To characterize the functional substitution of colon for the esophagus, we compared the electrogastrogram (EGG) maps and spectral frequencies and power of preoperative controls with patients who had undergone colonic replacement. Monopolar EGGs were recorded and spectrally analyzed at 27 locations on the thoracoabdominal surface. The spectral powers of five frequency groups were converted into EGG maps. In contrast to the epigastric concentrations of maximal power foci in a preoperative 3-cpm (cycles per minute) group, those of the colon replacement subjects seemed not to be concentrated in the epigastric region. Power in the 6-cpm colon replacement group were significantly greater and those in the 3-cpm colon replacement group were significantly less than those in the preoperative controls. Spectral frequencies in the 1- and 3-cpm colon replacement groups were significantly higher than those in the preoperative controls. The colonic and gastric EGG activities had 3-cpm, and probably 6-cpm in common. However, the colonic EGG activities were significantly different from the gastric EGG activities in frequency in the 3-cpm group, and in amplitude in both the 3- and 6-cpm groups. Thus, the replaced colon seems to preserve the original colonic EGG activity.


Antitumour activity of diallyl sulfide on polycyclic aromatic hydrocarbon-induced mouse skin carcinogenesis.
Diallyl sulfide (DAS), a major flavour component of garlic, is known to modulate xenobiotic metabolism and possess antitoxic, bactericidal, antineoplastic, hypolipidemic and hypocholesteromic effects. In the present study, the anticarcinogenic activity of DAS on a 7,12-dimethylbenzanthracene (DMBA)- or benzo[a]pyrene (B(a)P)-induced mouse skin model of carcinogenesis was evaluated. DAS was applied topically either 1 h prior to or 1 h after the administration of DMBA or B(a)P. A significant protection from neoplasia was observed in DAS- and DMBA/B(a)P-exposed animals when DAS was applied topically compared to the animals exposed only to DMBA/B(a)P. In the animals where DAS was applied 1 h prior to the application of DMBA, a lower magnitude of neoplasia was recorded in terms of the cumulative number of tumours and average number of tumours per mouse during the entire period of study (28 weeks) compared to the animals exposed to DAS 1 h later, while in B(a)P-exposed animals, the antitumorigenic potential of DAS was more evident in the mice treated with DAS 1 h after the B(a)P exposure compared to the animals treated with DAS 1 h prior to B(a)P. The antitumour activity of DAS was of a much higher magnitude in B(a)P-induced carcinogenesis in comparison to animals exposed to DMBA in terms of tumour incidence, cumulative number of tumours and average number of tumours per mouse. The results suggest that DAS has a protective effect in PAH-induced mouse skin carcinogenesis.


Sleep quality among Chinese elderly people: A population-based study.
Little was known about sleep quality among Chinese population. Using data from a population-based survey, we aimed to describe the prevalence of poor sleep quality among Chinese elderly people, to analyze factors associated with poor sleep quality, and to explore the relation between sleep quality and life quality as well as the risk for mental disorders. Data was derived from the Tianjin Mental Health Survey. The 19-item Pittsburg Sleep Quality Index was used to assess sleep quality. Life quality was evaluated using a Chinese version of Quality of Life Scale. To investigate the risk for mental disorders, the Chinese version of General Health Questionnaire was administered. Female gender, older age, low educational attainment, poor subject family economic status, and not having health insurance were significantly associated with elevated risk of poor sleep quality. Poor sleep quality was associated with poor life quality and increased odds of mental disorders. Further studies are warranted for the evaluation of effectiveness of screening tools for sleep problems and the generalization of early screening and diagnosing programs in the nation. Early screening is necessary and appropriate intervention programs should be conducted in the population with high risk.


In vitro activity of daptomycin in combination with low-dose colistin against a diverse collection of Gram-negative bacterial pathogens.
The activity of colistin in combination with daptomycin was assessed using 30 Gram-negative type strains and multidrug-resistant isolates with defined mechanisms of resistance. Daptomycin minimum inhibitory concentrations (MICs) were determined with and without sub-inhibitory concentrations of colistin. The activity of daptomycin was not affected with respect to Escherichia coli, Klebsiella pneumoniae or Pseudomonas aeruginosa. For colistin-susceptible Acinetobacter baumannii, sensitisation factors ranged from 8 to 128 (median 32), with the daptomycin MIC being reduced to the Clinical and Laboratory Standards Institute (CLSI) enterococci susceptibility breakpoint of 4 μg/ml for the ATCC 19606 type strain. A combination of daptomycin and colistin may be useful for the treatment of A. baumannii but not infections due to other Gram-negative species.


Sphingosine 1-phosphate lyase blockade elicits myogenic differentiation of murine myoblasts acting via Spns2/S1P2 receptor axis.
The bioactive sphingolipid sphingosine 1-phosphate (S1P) has emerged in the last three decades as main regulator of key cellular processes including cell proliferation, survival, migration and differentiation. A crucial role for this sphingolipid has been recognized in skeletal muscle cell biology both in vitro and in vivo. S1P lyase (SPL) is responsible for the irreversible degradation of S1P and together with sphingosine kinases, the S1P producing enzymes, regulates cellular S1P levels. In this study is clearly showed that the blockade of SPL by pharmacological or RNA interference approaches induces myogenic differentiation of C2C12 myoblasts. Moreover, down-regulation of the specific S1P transporter spinster homolog 2 (Spns2) abrogates myogenic differentiation brought about by SPL inhibition or down-regulation, pointing at a role of extracellular S1P in the pro-myogenic action induced by SPL blockade. Furthermore, also S1P2 receptor down-regulation was found to abrogate the pro-myogenic effect of SPL blockade. These results provide further proof that inside-out S1P signaling is critically implicated in skeletal muscle biology and provide support to the concept that the specific targeting of SPL could represent an exploitable strategy to treat skeletal muscle disorders.


Which Bone Mineral Density Measure Offers a More Reliable Prediction of Mechanical Complications in Adult Spinal Deformity Surgery: Hounsfield Units or DEXA Scan?
In patients undergoing adult spinal deformity (ASD) surgery, we sought to: (1) determine the relationship between dual-energy x-ray absorptiometry (DEXA)-measured bone mineral density (BMD), T-scores, and Hounsfield units (HU), and (2) compare the ability of DEXA-measured BMD, T-scores, and HU to predict mechanical complications and reoperations. A single-institution retrospective cohort study was undertaken for cases from 2013 to 2017. ≥5-level-fusion, sagittal/coronal deformity, and 2-year follow-up. Multivariable regression controlled for age, body mass index, receiving anabolic medications, and postoperative sagittal vertical axis and pelvic-incidence lumbar-lordosis mismatch. A subanalysis was performed for osteopenic patients (-1 < T-score < -2). Of 145 patients undergoing ASD surgery, 72 (49.6%) had both preoperative DEXA and computed tomography scans. Mean DEXA-measured BMD was 0.91 ± 0.52 g/cm2, mean T-score was -1.61 ± 1.03, and mean HU was 153.5 ± 52.8. While no correlation was found between DEXA-measured BMD and HU (r = 0.17, P = 0.144), T-score and HU had a weakly positive correlation (r = 0.31, P = 0.007). Mechanical complications occurred in 48 (66.7%) patients, including 27 (37.5%) proximal junctional kyphosis (PJK), 1 (1.4%) distal junctional kyphosis, 5 (6.9%) implant failure, 30 (41.7%) rod fracture/pseudarthrosis, 42 (58.3%) reoperations, and 16 (22.2%) reoperations due to PJK. No association was found between DEXA-measured BMD or T-scores with mechanical complications or reoperations. While univariate regression showed a significant association between lower HU and PJK (OR 0.98, 95%CI 0.97-0.99, P = 0.011), the significance was lost after multivariable analysis. When considering osteopenic patients (n = 37), only DEXA-measured BMD was an independent risk factor for PJK (OR 0.01, 95%CI 0.00-0.09, P = 0.017), with a threshold of 0.82 g/cm2 (AUC 0.70, 95%CI 0.53-0.84, P = 0.019). Poor correlation was found between the 3 BMD modalities. DEXA-measured BMD may be superior to T-scores and HU in predicting PJK among patients with osteopenia with a threshold of BMD <0.82 g/cm2.


Relationship between radiologist training level and radiation exposure for therapeutic hip injections.
The aim of this study was to determine the relationship between the level of training of the performing radiologist and patient radiation exposure, as measured by fluoroscopy time (FT) and dose area product (DAP), during fluoroscopic guided hip injections. This IRB-approved retrospective review of our institutional radiology report database identified all fluoroscopic guided hip injections performed between August 1st, 2012 and May 30th, 2015. Performing radiologists were divided into groups based on their level of training at the time of the procedure: first-year residents (R1), second-year residents (R2), third-year residents (R3), fourth-year residents (R4), staff fellowship trained musculoskeletal radiologists, and non-musculoskeletal radiologists. A mixed effects model was applied, using level of training as an independent predictor to model FT and DAP separately. There were 20, 19, 18 and 20 residents in the R1, R2, R3 and R4 groups, respectively. There were 5 performers in the non-MSK radiologist group, and 7 in the MSK radiologist group. A total of 1362 hip injection procedures met the inclusion criteria and were included in the analysis. The overall average FT was 26s (±17SD) and overall average DAP was 20.4uGy*m2 (±22.3SD). The mixed effects model showed no statistically significant difference between the groups, for FT (p=0.31) and for DAP (p=0.82). There is no association between radiologist level of training and patient radiation exposure for fluoroscopic hip injections. Resident trainees maintain radiation exposure to patients at levels comparable to their more experienced colleagues.


Identification of cis-acting promoter elements that support expression of membrane-type 1 matrix metalloproteinase (MT1-MMP) in v-src transformed Madin-Darby canine kidney cells.
Membrane-type 1 matrix metalloproteinase (MT1-MMP) expressed in tumor cells is believed to be important for the pericellular degradation of extracellular matrices during invasion and metastasis. To analyze the mechanism by which MT1-MMP becomes expressed in cancer cells, we assessed the MT1-MMP promoter region for the presence of cis-acting promoter elements that support transcription in transformed cells. Our tumor model consisted of Madin-Darby canine kidney (MDCK) cells transformed by v-src (src4 cells). MT1-MMP mRNA was only faintly detected in parental cells but was strongly expressed in the src4 cells. In parallel, src4 cells invaded into collagen gels, whereas MDCK cells did not. When MDCK and src4 cells were transiently transfected with a plasmid containing of -3000 to -99 nt from the upstream region of the MT1-MMP gene, the promoter activity was 2.6-fold higher in src4 cells than in MDCK cells. Furthermore, the region between -399 and -356 nt was found to contain the src4-specific enhancer element(s). Tandem Sp1 binding sites were also found to be essential in promoting transcription. An Egr-1 site that partially overlaps with the Sp1 sites was found to cooperate with the src4-specific enhancer and to also contribute weakly to the basal promoter activity. The presence of transcription factors that bind to the src4-specific enhancer site was detected by mobility-shift assays in src4 cell nuclear extracts but only weakly in MDCK extracts. Thus, we have identified a novel enhancer element that acts specifically in the transformed cells to enhance MT1-MMP expression.


Maturation stage enamel malformations in Amtn and Klk4 null mice.
Amelotin (AMTN) and kallikrein-4 (KLK4) are secreted proteins specialized for enamel biomineralization. We characterized enamel from wild-type, Amtn(-/-), Klk4(-/-), Amtn(+/-)Klk4(+/-) and Amtn(-/-)Klk4(-/-) mice to gain insights into AMTN and KLK4 functions during amelogenesis. All of the null mice were healthy and fertile. The mandibular incisors in Amtn(-/-), Klk4(-/-) and Amtn(-/-)Klk4(-/-) mice were chalky-white and chipped. No abnormalities except in enamel were observed, and no significant differences were detected in enamel thickness or volume, or in rod decussation. Micro-computed tomography (μCT) maximum intensity projections localized the onset of enamel maturation in wild-type incisors distal to the first molar, but mesial to this position in Amtn(-/-), Klk4(-/-) and Amtn(-/-)Klk4(-/-) mice, demonstrating a delay in enamel maturation in Amtn(-/-) incisors. Micro-CT detected significantly reduced enamel mineral density (2.5 and 2.4gHA/cm(3)) in the Klk4(-/-) and Amtn(-/-)Klk4(-/-) mice respectively, compared with wild-type enamel (3.1gHA/cm(3)). Backscatter scanning electron microscopy showed that mineral density progressively diminished with enamel depth in the Klk4(-/-) and Amtn(-/-)Klk4(-/-) mice. The Knoop hardness of the Amtn(-/-) outer enamel was significantly reduced relative to the wild-type and was not as hard as the middle or inner enamel. Klk4(-/-) enamel hardness was significantly reduced at all levels, but the outer enamel was significantly harder than the inner and middle enamel. Thus the hardness patterns of the Amtn(-/-) and Klk4(-/-) mice were distinctly different, while the Amtn(-/-)Klk4(-/-) outer enamel was not as hard as in the Amtn(-/-) and Klk4(-/-) mice. We conclude that AMTN and KLK4 function independently, but are both necessary for proper enamel maturation.


Psychological Traits in Patients Waiting for Total Knee Arthroplasty. A Cross-sectional Study.
To determine the prevalence of psychiatric distress in an Australian patient population waiting for total knee arthroplasty and to assess the associations between psychological distress and other baseline characteristics. This is a cross-sectional study of prospectively collected data from patients with knee arthritis participating in the New South Wales Osteoarthritis Chronic Care Program. Participants were divided into 2 groups based on the Depression Anxiety and Stress Score. The prevalence of psychological distress was compared to reported prevalence in the Australian community. Between-group comparisons of a number of variables were made, including demographic and socioeconomic data, health and psychiatric data, patient-reported knee and quality-of-life scores, and measured function. Data from 3103 patients were available and 2809 patients were eligible for inclusion, with 1740 (62%) females. Mean patient age was 68 years, and mean body mass index was 33.8. We found 753 (26.8%, standard deviation: 0.44) of participants reported psychological distress. Those reporting distress were more likely to be female (P = .025), younger (66.6 vs 68.8 years, P < .001), with a higher body mass index (34.5 vs 32.7, P < .001), and had more comorbidities (3.4 vs 2.8, P < .001). The distressed group had significantly worse knee pain (visual analog scale: 7.1 vs 6.2, P < .001; Knee Injury and Osteoarthritis Score pain component: 28.9 vs 41.2, P < .001), knee function, and general quality-of-health scores (P < .001). Psychological health is an important and often overlooked predictor of severity of symptoms and dysfunction in patients with knee arthritis. As it can affect the outcome for surgery, psychological health should be better assessed and treated before surgery to help reduce preoperative dysfunction and improve postoperative outcomes of total knee arthroplasty.


Synthesis of N-pyrimidinyl-2-phenoxyacetamides as adenosine A2A receptor antagonists.
A series of N-pyrimidinyl-2-phenoxyacetamide adenosine A(2A) antagonists is described. SAR studies led to compound 14 with excellent potency (K(i) = 0.4 nM), selectivity (A(1)/A(2A) > 100), and efficacy (MED 10 mg/kg p.o.) in the rat haloperidol-induced catalepsy model for Parkinson's disease.


In silico and in vitro screening of small molecule Inhibitors against SYT-SSX1 fusion protein in synovial sarcoma.
Synovial sarcoma (SS) is characterized by a tumour specific chromosomal translocation t(X;18) (p11;q11) which results in the formation of SYT-SSX1 fusion protein. This fusion protein represents a clear therapeutic target and molecules specifically targeting SYT-SSX1 fusion protein are currently not available. In this study, SYT-SSX1 fusion protein sequence was retrieved from Uniprot and 3D structure was generated using I-TASSER modeling program. A structure based computational screening approach has been employed using Glide docking software to identify potential SYT-SSX1 small molecule inhibitors that bind to the junction region of the fusion protein. The obtained inhibitors were further filtered based on the docking score and ADME/T properties. Ten best fit compounds were chosen for in vitro studies. The anti-proliferative activities of these 10 compounds were screened in Yamato, ASKA (carries SYT-SSX1 fusion protein) and other sarcoma cell lines such as A673, 143B to understand the specificity of inhibition of the chosen compounds. The in vitro activity was compared against HEK293 cell lines. The compound 5-fluoro-3-(1-phenyl-1H-tetraazol-5-yl)-1H-indole (FPTI) was found to be selectively cytotoxic in synovial sarcoma cell lines (Yamato and ASKA) and this compound also showed insignificant anti proliferative activity on other cell lines. Further, target gene expression study confirmed that FPTI treatment down-regulated SYT-SSX1 and modulated its downstream target genes. Cell cycle analysis revealed the involvement of an apoptotic mechanism of cell death. Further experimental validations may elucidate the therapeutic potentials of FPTI against SYT-SSX1 fusion protein.


Sensory Tricks in Pantothenate Kinase-Associated Neurodegeneration: Video-Analysis of 43 Patients.
Sensory tricks are a classic hallmark of primary dystonia and result in specific maneuvers that temporarily improve dystonic posture or movement. Pantothenate kinase-associated neurodegeneration (PKAN) is a progressive neurological disorder that courses with prominent dystonia. Although previously described, sensory tricks are considered to be rare in PKAN. We reviewed videotaped motor examinations of 43 genetically confirmed patients with PKAN in order to identify and classify sensory tricks. All patients presented some feature of dystonia. Eighteen (42%) had one or more well-structured sensory tricks. Twelve different sensory tricks were identified, eight typical and four atypical (forcible motor): four in cervical dystonia, four in limb dystonia, three in oromandibular dystonia, and one in blepharospasm. A characteristic forcible motor maneuver for oromandibular dystonia (previously described as the "mantis sign") was present in 8 patients. Sensory tricks are common in PKAN, particularly for oromandibular dystonia. The mantis sign may be a useful clue for the diagnosis.


Effect of operating parameters on desulphurization of mine tailings by froth flotation.
A site-specific study is carried out to assess the suitability of froth flotation for desulphurization of reactive mine tailings at the Musselwhite Mine, Northern Ontario, Canada, to prevent acid mine drainage (AMD). The results from pilot scale flotation tests on an Outokumpu flotation unit are presented, which confirm that froth flotation is effective to reduce sulphide contents of tailings. The factors affecting the treatment effectiveness, such as the froth depth, air flow rate, pulp density and impeller speed are studied. The sulphur recoveries after 0.25, 0.5, 1, 2, 5, and 12 min of flotation time are fitted to a second-order kinetic model. It is found that the second order rate constant, k(2) is negatively correlated with the froth depth and positively correlated with the air flow rate. Based on the data presented in this study; the maximum recovery of total sulphur was achieved when the operational parameters were set to the froth depth of 5 cm, air flow rate 125 L/min, impeller speed 1300 rpm and pulp density 35%.


Prevalence of Congenital Coronary Artery Anomalies and Variants in 2697 Consecutive Patients Using 64-Detector Row Coronary CTAngiography.
Coronary artery anomalies are not common, but could be very serious. This study determines the frequency of coronary anomalies and normal variants by multi-detector-row computed tomography (MDCT). The results of cardiac MDCT study in 2697 consecutive patients were analyzed retrospectively. Acquisition was performed by a 64-detector row CT machine. Imaging results were assessed by experienced radiologists. Myocardial bridging was by far the most frequent coronary variant (n = 576, 21.3%). Eighty-three subjects (3.1%) showed other coronary anomalies and variants. Anomalies of origination and course of the left main coronary artery (LMCA) were detected in 1.09% of the subjects. The frequency of these anomalies in the right coronary artery (RCA), left circumflex artery (LCx), left anterior descending artery (LAD), posterior descending artery (PDA) and obtuse marginal (OM) artery were 1.24%, 0.33%, 0.1%, 0.07% and 0.03%, respectively. The single coronary pattern was seen in 0.18% and coronary fistulas in 0.07%. Based on the fact that coronary CT-angiography using MDCT can display different coronary anomalies, this study shows similar results to other reports on the subject. Future advances in the performance of CT machines will further improve the quality of CT-based cardiac imaging.


New developments in instrumentation for vitreous surgery.
The paper describes new instruments for cutting of vitreous strands (vibrotome) and for removal of vitreous tissue (hyalophage). In addition, a motorized stereotaxic micromanipulator is presented.


Effects of structured involvement of the primary care team versus standard care after a cancer diagnosis on patient satisfaction and healthcare use: the GRIP randomised controlled trial.
The growing number of cancer survivors and treatment possibilities call for more personalised and integrated cancer care. Primary care seems well positioned to support this. We aimed to assess the effects of structured follow-up of a primary care team after a cancer diagnosis. We performed a multicentre randomised controlled trial enrolling patients curatively treated for breast, lung, colorectal, gynaecologic cancer or melanoma. In addition to usual cancer care in the control group, patients randomized to intervention were offered a "Time Out consultation" (TOC) with the general practitioner (GP) after diagnosis, and subsequent follow-up during and after treatment by a home care oncology nurse (HON). Primary outcomes were patient satisfaction with care (questionnaire: EORTC-INPATSAT-32) and healthcare utilisation. Intention-to-treat linear mixed regression analyses were used for satisfaction with care and other continuous outcome variables. The difference in healthcare utilisation for categorical data was calculated with a Pearson Chi-Square or a Fisher exact test and count data (none versus any) with a log-binomial regression. We included 154 patients (control n = 77, intervention n = 77) who were mostly female (75%), mainly diagnosed with breast cancer (51%), and had a mean age of 61 (SD ± 11.9) years. 81% of the intervention patients had a TOC and 68% had HON contact. Satisfaction with care was high (8 out of 10) in both study groups. At 3 months after treatment, GP satisfaction was significantly lower in the intervention group on 3 of 6 subscales, i.e., quality (- 14.2 (95%CI -27.0;-1.3)), availability (- 15,9 (- 29.1;-2.6)) and information provision (- 15.2 (- 29.1;-1.4)). Patients in the intervention group visited the GP practice and the emergency department more often ((RR 1.3 (1.0;1.7) and 1.70 (1.0;2.8)), respectively). In conclusion, the GRIP intervention, which was designed to involve the primary care team during and after cancer treatment, increased the number of primary healthcare contacts. However, it did not improve patient satisfaction with care and it increased emergency department visits. As the high uptake of the intervention suggests a need of patients, future research should focus on optimizing the design and implementation of the intervention. GRIP is retrospectively (21/06/2016) registered in the 'Netherlands Trial Register' (NTR5909).


Clinical characteristics and positive airway pressure adherence among elderly European sleep apnoea patients from the ESADA cohort.
The prevalence of obstructive sleep apnoea (OSA) is growing as the population is ageing. However, data on the clinical characteristics of elderly patients with OSA and their adherence to positive airway pressure (PAP) treatment are scarce. Data from 23 418 30-79-year-old OSA patients prospectively collected into the ESADA database during 2007-2019 were analysed. Information on PAP use (h·day-1) in association with a first follow-up visit was available for 6547 patients. The data was analysed according to 10-year age groups. The oldest age group was less obese, less sleepy and had a lower apnoea-hypopnoea index (AHI) compared with middle-aged patients. The insomnia phenotype of OSA was more prevalent in the oldest age group than in the middle-aged group (36%, 95% CI 34-38 versus 26%, 95% CI 24-27, p<0.001). The 70-79-year-old group adhered to PAP therapy equally well as the younger age groups with a mean PAP use of 5.59 h·day-1 (95% CI 5.44-5.75). PAP adherence did not differ between clinical phenotypes based on subjective daytime sleepiness and sleep complaints suggestive of insomnia in the oldest age group. A higher score on the Clinical Global Impression Severity (CGI-S) scale predicted poorer PAP adherence. The elderly patient group was less obese, less sleepy, had more insomnia symptoms and less severe OSA, but were rated to be more ill compared with the middle-aged patients. Elderly patients with OSA adhered to PAP therapy equally well as middle-aged patients. Low global functioning (measured by CGI-S) in the elderly patient predicted poorer PAP adherence.


The relationship between ochratoxin A and blood pressure in adolescents.
Ochratoxin A (OTA) is a chemical produced by some fungal species, and although its toxic effects have been shown in many animal studies, there are limited studies in humans. We aimed to examine the relationship between OTA and hypertension. 50 newly diagnosed hypertensive patients and 33 healthy individuals aged between 12 and 14 were included in the study. Anthropometric measurements, blood pressure measurements, complete blood count, blood biochemical parameters, urine lead level and urine OTA level were measured. OTA was detected in the urine samples of 90.9% of the control group, 100% of the hypertensive group and 85.7% of the obese+hypertensive group. Median urinary OTA was 32.9 ng/g creatinine for hypertensive group, 32.2 ng/g creatinine for hypertensive+obese group, 18.8 ng/g creatinine for the control group. Multivariate logistic regression analysis revealed a positive association between last quartile of urinary OTA level and being hypertensive [AOR:5.93 (95%CI: 1.27-27.61)] in adolescents without obesity. Hypertensive cases could be evaluated for OTA exposure in further studies.


Immune-associated pivotal biomarkers identification and competing endogenous RNA network construction in post-operative atrial fibrillation by comprehensive bioinformatics and machine learning strategies.
Atrial fibrillation (AF) is the most common arrhythmia. Previous studies mainly focused on identifying potential diagnostic biomarkers and treatment strategies for AF, while few studies concentrated on post-operative AF (POAF), particularly using bioinformatics analysis and machine learning algorithms. Therefore, our study aimed to identify immune-associated genes and provide the competing endogenous RNA (ceRNA) network for POAF. Three GSE datasets were downloaded from the GEO database, and we used a variety of bioinformatics strategies and machine learning algorithms to discover candidate hub genes. These techniques included identifying differentially expressed genes (DEGs) and circRNAs (DECs), building protein-protein interaction networks, selecting common genes, and filtering candidate hub genes via three machine learning algorithms. To assess the diagnostic value, we then created the nomogram and receiver operating curve (ROC). MiRNAs targeting DEGs and DECs were predicted using five tools and the competing endogenous RNA (ceRNA) network was built. Moreover, we performed the immune cell infiltration analysis to better elucidate the regulation of immune cells in POAF. We identified 234 DEGs (82 up-regulated and 152 down-regulated) of POAF via Limma, 75 node genes were visualized via PPI network, which were mainly enriched in immune regulation. 15 common genes were selected using three CytoHubba algorithms. Following machine learning selection, the nomogram was created based on the four candidate hub genes. The area under curve (AUC) of the nomogram and individual gene were all over 0.75, showing the ideal diagnostic value. The dysregulation of macrophages may be critical in POAF pathogenesis. A novel circ_0007738 was discovered in POAF and the ceRNA network was eventually built. We identified four immune-associated candidate hub genes (C1QA, C1R, MET, and SDC4) for POAF diagnosis through the creation of a nomogram and evaluation of its diagnostic value. The modulation of macrophages and the ceRNA network may represent further therapy methods.


Synthesis and NMR spectral assignments of novel nitrogen and sulfur heterocyclic compounds.
Synthesis and NMR spectral studies of multidentate N and S heterocycles, 1,3,5-tris(N-methylbenzimidazolyl)benzene, 1,3,5-tris(benzimidazolyl)benzene,1,3,5-tris(benzothiazolyl) benzene, 2,2'-bipyridine 3,3'-bis(benzothiazolyl)benzene and 1,2,4,5-tetrakis(benzothiazolyl) benzene have been carried out. 2D (1)H-(1)H PFG-COSY as well as (1)H-(13)C single and multiple bond correlated (2D GRASP-HSQC and GRASP-HMBC) experiments have been employed to characterize the compounds. 1D NOE experiments have been useful in understanding the structure of 1,3,5-tris(N-methylenzimidazolyl)benzene.


Paraplane analysis from precordial three-dimensional echocardiographic data sets for rapid and accurate quantification of left ventricular volume and function: a comparison with magnetic resonance imaging.
Three-dimensional echocardiography (3DE) calculates left ventricular volumes (LVV) and ejection fraction (EF) without geometric assumptions, but prolonged analysis time limits its routine use. This study was designed to validate a modified 3DE method for rapid and accurate LVV and EF calculation compared with magnetic resonance imaging (MRI). Forty subjects included 15 normal volunteers (group A) and 25 patients with segmental wall motion abnormalities and global hypokinesis caused by ischemic heart disease (group B) who underwent 3DE with precordial rotational acquisition technique (2-degree interval with electrocardiographic and respiratory gating) and MRI at 0.5 T, electrocardiogram (ECG)-triggered multislice multiphase T1-weighted fast field echo. End-diastolic and end-systolic LVV and EF were calculated from both techniques with Simpson's rule by manual endocardial tracing of equidistant parallel left ventricular short-axis slices. Slicing from the 3DE data sets were done by both 2.9-mm slice thickness (method 3DE-A) and by 8 equidistant short-axis slices (method 3DE-B); for MRI analysis, 9-mm slice thickness was used. Analysis time required for manual endocardial tracing of end-diastolic and end-systolic short-axis slices was 10 minutes for the 3DE-B method compared with 40 minutes by the 3DE-A method. For all 40 subjects the mean +/- SD of end-diastolic LVV (mL) were 181 +/- 76, 179 +/- 73, and 182 +/- 76; for end-systolic LVV (mL), 120 +/- 76, 120 +/- 75, and 122 +/- 77; and for EF (%), 39 +/- 18, 38 +/- 18, and 38 +/- 18 for MRI, 3DE-A, and 3DE-B methods, respectively. The differences between 3DE-A and 3DE-B with MRI for calculating end-diastolic and end-systolic LVV and EF were not significant for the whole group of subjects as well as for the subgroups. The 3DE-B method had excellent correlation and close limits of agreement with MRI for calculating end-diastolic and end-systolic LVV and EF: r = 0.98 (-1.3 +/- 26.6), 0.99 (-1.6 +/- 21. 2), and 0.99 (0.2 +/- 5.2), respectively. The correlation between 3DE-A and MRI were r = 0.97, 0.98, and 0.98, and the limits of agreement were -1.4 +/- 36, -0.6 +/- 26, and 0.6 +/- 8 for calculating end-diastolic and end-systolic LVV and EF, respectively. In addition, excellent correlation and close limits of agreement between 3DE-A and 3DE-B with MRI for LVV and EF calculation was also found for the subgroups. Intraobserver and interobserver variability (SEE) of MRI for calculating end-diastolic and end-systolic LVV and EF were 6.3, 4.7, and 2.1; and 13.6, 11.5, and 4.7; respectively, whereas that for 3DE-B were 3.1, 4.4, and 2.2; and 6.2, 3.8, and 3. 6; respectively. Comparable observer variability was also found for the A and B subgroups. The 3DE-A and 3DE-B methods have excellent correlation and close limits of agreement with MRI for calculating LVV and EF in both normal subjects and cardiac patients. The 3DE-B method by paraplane analysis with 8 equidistant short-axis slices has observer variability similar to MRI and reduces the 3DE analysis time to 10 minutes, therefore offering a rapid, reproducible, and accurate method for LVV and EF calculation.


Rotational dynamics of 4-aminobutyrate aminotransferase.
The fluorescence dye 1-anilinonaphthalene-8-sulfonate (ANS) was used as a probe of non-polar binding sites in 4-aminobutyrate aminotransferase. ANS binds to a single binding site of the dimeric protein with a Kd of 6 microM. Nanosecond emission anisotropy measurements were performed on the ANS-enzyme in an effort to detect independent rotation of the subunits in the native enzyme. The observed rotational correlation time (phi = 65 ns) corresponds to the rotation of a rather rigid dimeric structure. The microenvironment surrounding the natural probe pyridoxal-5-P covalently bound to the dimeric structure was explored using 31P-NMR at 72.86 MHz. In the native enzyme, the pyridoxal-5-P 31P-chemical shift is pH-independent, indicating that the phosphate group is well protected from the solvent. The correlation time determined from the 31P-spectrum of the aminotransferase exceeds the value calculated for the hydrated spherical model (phi = 40 ns). It is concluded that the phosphate of the pyridoxal-5-P molecule is rigidly bound to the active site of 4-aminobutyrate aminotransferase.


Effect of MI-D, a new mesoionic compound, on energy-linked functions of rat liver mitochondria.
MI-D (4-phenyl-5-(4-nitro-cinnamoyl)-1,3,4-thiadiazolium-2-phenylami ne chloride), a new mesoionic compound, depressed the phosphorylation efficiency of liver mitochondria as deduced from an accentuated decrease of the respiratory control coefficient and ADP/O ratio. Analysis of segments of the respiratory chain suggested that the MI-D inhibition site is further on than complex I and between complexes II and III. The transmembrane electrical potential (delta psi) was collapsed dependent on MI-D concentration. ATPase activity was dramatically increased by MI-D in intact mitochondria, but inhibited in carbonylcyanide p-trifluoromethoxyphenylhydrazone (FCCP)-uncoupled mitochondria. These results suggest that MI-D acts as an uncoupler agent, a property closely related to its structural characteristics.


Features and preliminary results of the Dutch centre of the ERSPC (Rotterdam, the Netherlands).
To describe the preliminary results of the Dutch section of a large multicentre study of screening for prostate cancer, the European Randomized study of Screening for Prostate Cancer (ERSPC), initiated in the Netherlands and Belgium in 1991. After a series of five pilot studies which started in 1991, full-capacity screening started in 1994 with the use of a serum prostate-specific antigen (PSA) determination, a digital rectal examination (DRE) and transrectal ultrasonography (TRUS) as screening tests. Depending on the results and the screening protocol used, men were referred for further examination by sextant biopsies (extended with a seventh biopsy if TRUS showed abnormality). The protocols used, efficiency of the different screening tests, number of cancers detected in the pilot studies, initial screening round and preliminary results of the second screening round are described. After the pilot studies it became clear that a study of prostate cancer screening was feasible in the Rotterdam area. The screening protocol was workable and the recruitment rate acceptable (39.5%). An inventory of the population registries of Rotterdam and surrounding municipalities, and the known recruitment rate, made it clear that a contribution of 40000 men (aged 55-74 years) from the Dutch centre to the ERSPC was feasible. The initial screening round started in December 1993 and lasted until December 1999 (protocol 5-10). In all, 42376 men were randomized and 1014 cancers detected (5.1%). During this screening the protocol was simplified. After evaluating the different screening tests abnormal results of the DRE and TRUS were omitted as an indication for a sextant biopsy. Only a serum PSA level of > or = 3.0 ng/mL is now used as the indication. The second screening round started in December 1997 and continues. To December 2002, 9920 men were screened for the second time, 4 years after their initial screening visit. To date 446 cancers have been detected (4.5%); this round will last to December 2003. Further evaluation of the screening regimen and characteristics of the cancers detected are constantly assessed within the Dutch ERSPC. Meanwhile a third screening round has also been initiated, which will last to December 2007. A prostate cancer screening study of the projected magnitude is feasible in Rotterdam; the recruitment rate is acceptable and the screening tests well tolerated. The study has generated many scientific publications and will be of great value in determining whether prostate cancer screening should be part of general healthcare.


EFA6 activates Arf6 and participates in its targeting to the Flemming body during cytokinesis.
The small GTPase Arf6 is transiently associated with the ingressing cleavage furrow and subsequently targeted to the Flemming body during cytokinesis, suggesting its activation around the cleavage furrow. Here, we show that EFA6 (exchange factor for Arf6) localizes on the cleavage furrow through its PH domain. Time-lapse analysis showed that both EFA6 and Arf6 are transiently localized around the ingressing cleavage furrow, but only Arf6 is subsequently targeted to the Flemming body. Expression of an EFA6 mutant suppresses Arf6 recruitment onto the Flemming body. These results suggest that EFA6 participates in activation of Arf6 around the cleavage furrow during cytokinesis.


Bibliometric analysis of nanotechnology applied in oncology from 2002 to 2011.
Innovation in the last decade has endowed nanotechnology with an assortment of tools for drug delivery system, imaging, and sensing in cancer research. These rapidly emerging tools are indicative of a burgeoning field ready to expand into medical applications. The aim of this study is to analyze the applications of nanotechnology in oncology with bibliometric methods and evaluate development in this field. Literature search was performed using PubMed search engines with MeSH terms (all)--nanotechnology, nanomedicine, nanoparticle, nanocapsules, micellar systems, and oncology or cancer or neoplasms. Within 2,543 articles from 2002 to 2011 in over 50 medical magazines from over 30 countries, we did a series analysis on these articles' countries, keywords, and authors. Our results show that articles in nanotechnology in oncology are increasing year by year, especially in recent years. Quantity and quality of the articles are becoming more and influential. In the global research, the USA is leading in this field, accounting for half above of the whole articles, followed by countries like Japan, Germany, and France and also some emerging nations like China, in the second place, and India. Subjects like nanoparticles, tumor marker, and drug delivery are the common research focus. So, with more and more scientists' interests and attention drawn to this field, it is likely to make major breakthroughs in the coming years.


Specialty pharmacy integration and the role of an advanced certified pharmacy technician in prescription cannabidiol access.
Insurance requirements that limit access to prescription cannabidiol (CBD), an adjunct therapy for uncontrolled seizure disorders, may lead to treatment initiation delays. Integrated health-system specialty pharmacies (IHSSPs) use pharmacists and advance certified pharmacy technicians (CPhTs) to help navigate prescription CBD access requirements. Evaluate time from initial specialty pharmacy referral to prescription CBD shipment. This was a single-center, retrospective analysis of patients prescribed CBD from January 2019 to April 2020 by the outpatient neurology clinic and dispensed by the center's IHSSP. The primary outcome was the time to prescription CBD access, defined as days between the specialty pharmacy completing an initial patient assessment and first medication shipment. Secondary outcomes were percentage of patients requiring financial assistance and days between key steps in the access pathway. Data were collected from electronic health records and the specialty pharmacy patient management database. The CPhT was responsible for completing most portions of the access pathway under supervision of the clinical pharmacist. After screening, 136 patients were included: 50% male, 85% white, 60% insured by Medicaid, and median age 14 years (interquartile range [IQR] 9-21). The most common indication was Lennox-Gastaut syndrome (n = 117, 86%). Of the 129 patients (95%) who required a prior authorization (PA), 92% were approved (n = 119). Median time from initial assessment to first shipment was 7 days (IQR 4-13). Of patients for whom the CPhT helped obtain financial assistance (n = 14, 10%), all had $0 costs after assistance. Median times for secondary outcomes led by the CPhT in days were as follows: initial assessment completion to benefits investigation (BI) = 0 (IQR 0-0), BI to PA submission = 0 (IQR 0-0), and PA denial to appeal submission = 4 (IQR 1-7). IHSSP teams, particularly advanced CPhT roles, helped patients afford and initiate prescription CBD quickly.


Identification of a glycolysis-related miRNA Signature for Predicting Breast cancer Survival.
Breast cancer (BC) is a common type of cancer and has a poor prognosis. In this study, we collected the mRNA and miRNA expression profiles of BC patients were obtained from The Cancer Genome Atlas (TCGA) to explore a novel prognostic strategy for BC patients using bioinformatics tools. We found that six glycolysis-related miRNAs (GRmiRs, including hsa-mir-1247, hsa-mir148b, hsa-mir-133a-2, has-mir-1307, hsa-mir-195 and hsa-mir-1258) were correlated with prognosis of BC samples. The risk score model was established based on 6 prognosis-associated GRmiRs. The outcome of high risk group was significantly poorer. Cox regression analysis showed that risk score was an independent prognostic factor. Differentially expressed genes identified between high and low risk groups were mainly enriched in inflammation and immune-related signaling pathways. The proportion of infiltration of 12 kinds of immune cells in high and low risk groups were significantly different. Risk score was closely associated with many immune indexes. Multiple DEGRGs and miRNAs were associated with drugs. In conclusion, glycolysis-related miRNA signature effectively predicts BC prognosis.


Gaucher type I (Ashkenazi) disease: a new method for heterozygote detection using a novel fluorescent natural substrate.
A new acid beta-glucosidase assay for the detection of heterozygotes for Gaucher Type I disease has been developed using isolated lymphocytes as enzyme source and a novel fluorescent natural substrate, NBD-glucosyl ceramide. The procedure for optimal heterozygote discrimination was established by systematic evaluation of the effect of various solubilization techniques, detergent concentrations, pH, enzyme sources and artificial vs. natural substrates. A pilot screening study indicated the feasible application of this assay for heterozygote detection in the high-risk Ashkenazi Jewish population.


Bias varies for bioimpedance analysis and skinfold technique when stratifying collegiate male athletes' fat-free mass hydration levels.
This study evaluated the accuracy of bioimpedance analysis (BIA)- and skinfold (SF)-based body fat percentage estimates in collegiate athletes with varying fat-free mass (FFM) hydration levels. Subjects were evaluated as a whole (n = 63) and at FFM hydration levels of 64.00%-68.99% (FFM-HydrationL1; n = 37) and 69.00%-74.00% (FFM-HydrationL2; n = 26). Proportional bias was absent in the SF technique when stratifying FFM hydration levels. Contrarily, proportional bias was observed when using BIA for FFM-HydrationL1, but not in FFM-HydrationL2. Novelty Fat-free mass hydration levels impact BIA-based body fat estimates more than skinfold-based body fat.


Increased footwear comfort is associated with improved running economy - a systematic review and meta-analysis.
Footwear with or without custom foot orthotics have the potential to improve comfort, but the link with running performance needs further investigation. We systematically reviewed the association of footwear comfort on running economy in recreational runners. Nine electronic databases were searched from inception to March 2020. Eligible studies investigated both direct outcome measures of running performance (e.g. running speed) and/or physiological measures (e.g. running economy (RE)) alongside comfort for each footwear condition tested. Methodological quality was assessed using the "Effective Public Health Practice Project" (EPHPP). RE during submaximal running was the most common physiological outcome reported in 4 of the 6 eligible studies. The absolute difference in RE between the most and least comfortable footwear condition was computed, and meta-analysis was conducted using a random effect model. The most comfortable footwear is associated with a reduction in oxygen consumption (MD: -2.06 mL.kg-1.min-1, 95%CI: -3.71, -0.42, P = 0.01) while running at a set submaximal speed. There was no significant heterogeneity (I2 = 0%, P = 0.82). EPHPP quality assessment demonstrated weak quality of the studies, due to reporting bias and failing to disclose the psychometric properties of the outcome measures. It can be concluded with moderate certainty that improved RE in recreational athletes is associated with wearing more comfortable footwear compared to less comfortable footwear.HighlightsThis systematic review reports on the association of footwear comfort with running economy in recreational runners.Running economy during constant submaximal running is likely improved in recreational runners wearing more comfortable compared to less comfortable footwear.This finding is based on a meta-analysis, including four studies, showing a small but statistically significant decreased oxygen consumption at steady state speeds while wearing the most comfortable footwear.


[THE INFLUENCE OF THE MELATONIN ON THE CORRELATION BETWEEN THE INTENSITY OF THE ACCUMULATION OF THE OXIDATI-VE-MODIFIED PROTEINS CONTENT, ACTIVITY OF THE ANTIOXIDANT ENZYMES AND THE STATE OF PROTEOLYSIS IN THE BASAL NUCLEI OF THE BRAIN UNDER THE ACUTE HYPOXIA].
The effect of melatonin on the correlation between the intensity of the accumulation of the oxidative-modified protein content, activity of the antioxidant enzymes and the state of proteoly-sis in the basal nuclei (the nucleus caudatus, globus pallidus, nucleus accumbens, amigdaloid complex) of the brain under the conditions of acute hypoxia has been studied. Under the conditions of acute hypoxia in the basal nuclei an intensification of the protein peroxidation processes is observed, the activity of the antioxidant enzymes decreases, the intensity of the proteolysis increases. The injection of melatonin in a dose of 1 mg per kg of the body mass before the modeling of acute hypoxia results in the decreasing of protein peroxidation, increasing of the antioxidant enzyme activity and normalization of the parameters of proteolysis.


Berberine inhibits the growth of human colorectal adenocarcinoma in vitro and in vivo.
Berberine is an alkaloid isolated from the Chinese herbal medicine Huanglian, and has long been used as an antibiotic. Its antineoplastic properties were subsequently discovered in vitro. The purpose of this study was to investigate the effects of berberine on the growth of human colorectal carcinoma cells in vitro and in vivo. The results showed that berberine inhibited human colorectal adenocarcinoma (LoVo) cell growth in a time- and dose-dependent manner. A WST-1 assay showed that the IC50 value after 72 h was 40.79 ± 4.11 μM. Cell cycle analysis of 40 μM berberine-treated LoVo cells by flow cytometry showed accumulation of cells in the G2/M phase. The inhibition of LoVo cell growth by berberine was associated with the suppression of cyclin B1, cdc2, and cdc25c proteins. Berberine at a dose of 50 mg kg(-1) day(-1) showed inhibitory rates of 45.3% in a human colorectal adenocarcinoma xenograft in nude mice. The combination of berberine and 5-fluorouracil (5-FU) had a higher inhibitory rate (59.8%) than the berberine group (36.4%, P = 0.01), but no significant difference was observed between the 5-FU group (43.0%, P = 0.06) and the combination group. These results support the possibility that berberine may be useful as an alternative therapy for colorectal carcinoma.


Synthesis, spectral and solid state characterization of a new bioactive hydrazine bridged cyclic diphosphonium compound.
The facile preparation of a racemic hydrazine bridged diphosphonium compound possessing a ring system analogous to bicyclo[3.3.2]decane is reported. Although the reaction yield is low, the structure of the compound, which possesses an eight-membered ring, two phosphonium cationic centers, a biimino bridge, molecular chirality and two fused aromatic rings locked into roughly perpendicular planes is unusual. The compound displays substantial biological activity in the brine shrimp test and cleaves plasmid DNA.


Using photonic crystal enhanced fluorescence on quartz substrates to improve the sensitivity of DNA microarrays.
Gene expression analysis of low abundance genes remains difficult when DNA microarrays are performed on standard glass substrates. However, we have shown that by using photonic crystals (PC) made on quartz substrates, the fluorescence intensity of Cyanine-5 (Cy5) labeled microarray spots is greatly enhanced. In a 1-color microarray experiment studying gene expression of soybean cotyledon tissue, an average signal enhancement factor of 17.8× was observed on the PC. Furthermore, twice as many genes were detectable on these PCs as compared to glass. By improving the sensitivity of this fluorescent assay, low expression genes that were undetectable on glass were quantified on the PC.


Adult height in thalassaemia major without hormonal treatment.
Height and sitting height were measured in all the 28 patients (19 females and 9 males) of our Thalassaemic Centre who exhibited evidence of cessation of growth. Their average final stature, which had been spontaneously achieved at an age ranging from 15.0-21.5 years (mean 17.9), was slightly lower with respect to the mean standards for women (159.1 +/- 8.4 vs 160.0 +/- 6.0 cm) and men (167.8 +/- 6.4 vs 172.5 +/- 6.0 cm). Height deficiency exceeded 2 SD scores in only 3/28 patients. If compared to the familial target height, patients' stature fell within the familial range in 19/24 cases and in only one case was it below the lower limit. In the whole sample the patients' height was positively related to their familial target height (r = 0.72, P less than 0.0002) and also to mid-parental stature (r = 0.45, P less than 0.05). Heights of both females and males were superimposable on those of their own mothers and fathers respectively. Average values of either SH (76.4 +/- 3.8 vs 79.9 +/- 3.4 cm, 2 P less than 0.02) or SH/H ratio (48.9 +/- 2.5 vs 51.5 +/- 1.0, 2 P less than 0.005) were significantly lower in the female thalassaemics than in the sex-matched controls. Three females and one male showed subnormal values of both SH and SH/H ratio. Two out of these patients with eunochoid body proportions and another one with a decreased SH/H ratio have hypogonadotropic hypogonadism and are undergoing a chronic substitutive treatment with sexual hormones, which was instituted after the achievement of final stature.


Hu'po Anshen decoction promotes fracture healing in mice with traumatic brain injury through BMP2-COX2-ATF4 signaling pathway.
Hu'po Anshen decoction (HPASD), a traditional Chinese medicine used to treat concussion and fracture, could regulate the expression of bone morphogenetic protein 2 (BMP2). However, whether HPASD affects the fracture healing of traumatic brain injury (TBI) combined with a fracture through BMP2 and its downstream signals remains obscure. The chondrocyte-specific BMP2 conditional knockout mice and chondrocyte-specific cyclooxygenase-2 (COX2) overexpression mice were generated. BMP2 conditional knockout mice were treated with fracture surgery, fracture combined with TBI, or fracture combined with TBI followed by different doses of HPASD (2.4, 4.8, and 9.6 g/kg), respectively. TBI was induced by Feeney's weight-drop technique. The fracture callus formation and fracture sites were determined by X-ray, micro-CT, and histological analyses. The expressions of chondrocyte-, osteoblast-, and BMP2/COX2 signal-related targets were determined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blot assays. The specific absence of BMP2 in chondrocytes led to the prolonged formation of cartilage callus, a delay in the osteogenesis initiation and the downregulation of RUNX2, Smad1/5/9, EP4, ERK1/2, RSK2, ATF4. Overexpression of COX2 partially reverses the effects of chondrocyte-specific BMP2 knockout mice. HPASD promoted cartilage callus formation and osteogenesis initiation, as accompanied by upregulated expression levels of RUNX2, Smad1/5/9, EP4, ERK1/2, RSK2, and ATF4 in a time-dependent and concentration-dependent manner in chondrocyte-specific BMP2 knockout mice. Overall, our findings demonstrated that HPASD induced COX2 transcription through the BMP2-Smad1/5/9-RUNX2 axis, and then affected fracture healing through the COX2-mediated EP4-ERK1/2-RSK2-ATF4 axis.


GREB1 is a novel androgen-regulated gene required for prostate cancer growth.
Gene regulated in breast cancer 1 (GREB1) is a novel estrogen-regulated gene shown to play a pivotal role in hormone-stimulated breast cancer growth. GREB1 is expressed in the prostate and its putative promoter contains potential androgen receptor (AR) response elements. We investigated the effects of androgens on GREB1 expression and its role in androgen-dependent prostate cancer growth. Real-time PCR demonstrated high level GREB1 expression in benign prostatic hypertrophy (BPH), localized prostate cancer (L-PCa), and hormone refractory prostate cancer (HR-PCa). Androgen treatment of AR-positive prostate cancer cells induced dose-dependent GREB1 expression, which was blocked by anti-androgens. AR binding to the GREB1 promoter was confirmed by chromatin immunoprecipitation (ChIP) assays. Suppression of GREB1 by RNA interference blocked androgen-stimulated LNCaP cell proliferation. GREB1 is expressed in proliferating prostatic tissue and prostate cancer, is regulated by androgens, and suppression of GREB1 blocks androgen-induced growth suggesting GREB1 may be critically involved in prostate cancer proliferation.