New resistance-correlated saponins from the insect-resistant crucifer Barbarea vulgaris.
Isolation and characterization of plant constituents responsible for insect resistance are of the utmost importance for better understanding of insect-host plant interactions, for selection and breeding of resistant plant varieties, and for development of natural insecticides to be used in future sustainable agriculture and food production. In this study, 3-O-cellobiosyl-cochalic acid (1), 3-O-cellobiosyl-gypsogenin (3), and 3-O-cellobiosyl-4-epihederagenin (4) were isolated from the glabrous type of Barbarea vulgaris var. arcuata exhibiting resistance to the flea beetle Phyllotreta nemorum. In addition to the new constituents, 3-O-cellobiosyl-hederagenin (2), a known insect repellant, was identified. The structures were established by one- and/or two-dimensional homo- and heteronuclear NMR experiments acquired at 800 MHz and by fragmentation and high-resolution mass spectrometric analysis. Compounds 1, 3, and 4 are glycosides of cochalic acid, gypsogenin, and 4-epihederagenin, respectively, none of which have previously been identified in Brassicaceae. Compounds 3 and 4 have both recently been targeted as unidentified constituents exhibiting correlation with P. nemorum resistance, but this is the first report of their structures.


One-trial visual recognition in cats.
The ability of normal cats to perform delayed matching- and nonmatching-to-sample with trial-unique stimuli was investigated both in a modified Wisconsin General Testing Apparatus requiring manipulatory responses and in a Nencki-type testing room requiring locomotor responses. Cats trained in the WGTA learned the two tasks at about the same rate, on average, as that reported for monkeys. However, unlike monkeys, whose strong preference for novelty facilitates their learning of the nonmatching rule and retards their learning of the matching rule, the cats learned the two different rules at about the same rate, suggesting that cats do not share the monkey's strong preference for novelty. In contrast to their relatively rapid learning of the manipulatory versions of the two tasks, cats learned the locomotor versions only slowly or even failed to learn. Experimental analysis indicated that a major source of the cats' difficulty on these locomotor versions was interference from a strong tendency in the large testing room to use visuospatial strategies. Nevertheless, once the matching or nonmatching rule was learned at short delays, whether in the WGTA or the testing room, the cats performed at criterion levels without further training even at delays of 10 minutes, indicating that this species, like monkeys, has a highly developed long-term recognition memory ability.


Do non-HLA genes influence development of persistent islet autoimmunity and type 1 diabetes in children with high-risk HLA-DR,DQ genotypes?
Specific alleles of non-HLA genes INS, CTLA-4, and PTPN22 have been associated with type 1 diabetes. We examined whether some of these alleles influence development of islet autoimmunity or progression from persistent islet autoimmunity to type 1 diabetes in children with high-risk HLA-DR,DQ genotypes. Since 1993, the Diabetes Autoimmunity Study in the Young (DAISY) has followed 2,449 young children carrying HLA-DR,DQ genotypes associated with type 1 diabetes. Of those, 112 have developed islet autoimmunity (persistent autoantibodies to insulin, GAD65, and/or IA-2), and 47 of these have progressed to type 1 diabetes. The influence of polymorphisms of INS(-23Hph1), CTLA-4(T17A), and PTPN22(R620W) on development of persistent islet autoimmunity and progression to type 1 diabetes was evaluated by parametric models and by survival analyses. PTPN22(R620W) allele T was associated with development of persistent islet autoimmunity (hazard ratio 1.83 [95% CI 1.27-2.63]) controlling for ethnicity, presence of HLA-DR3/4,DQB1*0302, and having a first-degree relative with type 1 diabetes. Survival analyses showed a significantly (P = 0.002) higher risk of persistent islet autoimmunity by age 10 years for the TT genotype (27.3%) than for the CT or CC genotype (7.9 and 5.3%, respectively). Cumulative risk of persistent islet autoimmunity was slightly higher (P = 0.02) for the INS(-23Hph1) AA genotype (7.8%) than for the AT or TT genotype (4.2 and 6.4% risk by age 10 years, respectively). Whereas the HLA-DR3/4,DQB1*0302 genotype had a dramatic influence on both development of islet autoimmunity and progression to type 1 diabetes, the PTPN22(R620W) T allele significantly influences progression to persistent islet autoimmunity in the DAISY cohort.


Chronic sulpiride treatment produces supersensitivity of striatal adenylate cyclase to dopamine in sexually immature or adult castrated rats.
Sulpiride, a substituted benzamide antipsychotic drug, is considered to be a selective antagonist at dopamine D-2 receptors, largely because it does not inhibit dopamine-stimulated adenylate cyclase activity. It was found that sulpiride in vitro can block dopamine-stimulated adenylate cyclase activity in rat striatum from sexually immature or adult castrated male rats. Chronic treatment with sulpiride (20 mg/kg i.p. twice daily for 15 days) resulted in supersensitivity of dopamine-stimulated adenylate cyclase activity in striatum from sexually immature (3-4 weeks old) as well as adult castrated rats. The apparent Ka was decreased 4-fold in the sulpiride-treated animals, whereas the apparent Vmax remained unchanged. This treatment did not alter dopamine-stimulated adenylate cyclase activity in the striatum from adult male rats. Acute treatment with sulpiride was slightly inhibitory to dopamine-stimulated adenylate cyclase in striatum from the immature rat, suggesting that the supersensitivity after repeated injections could be a compensatory increase. In in vitro studies it was found that sulpiride at nanomolar levels could inhibit the ability of low concentrations of dopamine to stimulate adenylate cyclase activity in striatal particulate fractions from sexually immature or adult castrated rats. Sulpiride did not affect dopamine-stimulated adenylate cyclase in the adult rat striatum. Our results indicate that sulpiride can affect dopamine-stimulated adenylate cyclase activity in animals that are lacking testosterone, or perhaps estrogen. This suggests that sex hormones could regulate the sensitivity and pharmacological profile of dopamine receptors for their ligands.


Perceived environment and physical activity: a meta-analysis of selected environmental characteristics.
Several narrative reviews have been conducted on the literature examining environmental correlates of physical activity (PA). To date these reviews have been unable to provide definitive summaries of observed associations. This study utilizes meta-analytical techniques to calculate summaries of associations between selected environmental characteristics and PA. Published studies were identified from electronic databases and searches of personal files. Studies were examined to determine the environmental constructs most frequently studied. Included studies (N = 16) examined at least one identified construct and determined associations between perceived environmental constructs and PA using logistic regression. Data were analyzed separately for crude and adjusted ORs using general-variance based fixed effect models. No significant associations emerged between environmental characteristics and PA using crude OR. The perceived presence of PA facilities (OR 1.20, 95% 1.06-1.34), sidewalks (OR 1.23, 95% 1.13-1.32), shops and services (OR 1.30, 95% 1.14-1.46) and perceiving traffic not to be a problem (OR 1.22, 95% 1.08-1.37) were positively associated with activity using adjusted ORs. Variance in PA accounted for by significant associations ranged from 4% (heavy traffic not a problem) to 7% (presence of shops and services). Results of the meta-analysis support the relevance of perceived environmental characteristics for understanding population PA. These results should encourage the use of comprehensive ecological models that incorporate variables beyond basic demographic information.


Serial assessment of left ventricular performance following coronary artery bypass grafting. Early postoperative results with myocardial protection afforded by multidose hypothermic potassium crystalloid cardioplegia.
Forty patients who recently underwent coronary artery bypass graft (CABG) operations had serial hemodynamic and scintigraphic studies. Multidose hypothermic potassium crystalloid cardioplegia was used for myocardial protection and newer techniques in anesthetic management and perioperative patient care were also employed. The method of equilibrium cardiac gated blood pool (GBP) scintigraphy was used to obtain perioperative changes in global ejection fraction (EF) and regional wall motion (RWM). Ninety percent of patients displayed a decrease in EF 2 hours postoperatively when compared to their preoperative values. This change was also associated with a fall in cardiac index (CI) and left ventricular stroke work index (LVSWI). Twenty-four hours postoperatively, EF and CI recovered to preoperative levels, but LVSWI remained depressed. Seven days postoperatively, global EF had improved to a value greater than the preoperative one (50% +/- 3% versus 57% +/- 4%, p < 0.05). Perioperative changes in RWM followed the same pattern as EF, but recovery in this index of regional contractility was faster than EF, since maximal improvement was observed 24 hours postoperatively. Thus transient left ventricular dysfunction is common immediately after CABG, but recent advances in myocardial protection and perioperative management are associated with short-term increases in regional and global left ventricular function documented by noninvasive GBP imaging.


EpCAM and APN expression in combination with γ-H2AX as biomarkers for detecting hepatocarcinogens in rats.
The phosphorylated form of histone H2AX (γ-H2AX) serves as a commonly utilized biomarker for DNA damage. Based on our previous findings, which demonstrated the formation of γ-H2AX foci as a reliable biomarker for detecting bladder carcinogens in repeated dose 28-day study in rats, we hypothesized that γ-H2AX could also function as a biomarker for detecting hepatocarcinogens. However, we found that γ-H2AX foci formation was not effectively induced by hepatocarcinogens that did not stimulate hepatocyte proliferation. Therefore, we explored alternative biomarkers to detect chemical hepatocarcinogenicity and discovered increased expressions of epithelial cell adhesion molecule (EpCAM/CD326)- and aminopeptidase N (APN/CD13) in the hepatocytes of rats administered various hepatocarcinogens. Significant increases in EpCAM- and APN-positive hepatocytes were observed for eight and five of the 10 hepatocarcinogens, respectively. Notably, five and two of them, respectively, were negative for γ-H2AX foci. These results highlight the potential of EpCAM and APN as useful biomarkers in combination with γ-H2AX for the detection of chemical hepatocarcinogenicity.


Association of the type 1 inositol (1,4,5)-trisphosphate receptor with 4.1N protein in neurons.
The type 1 inositol (1,4,5)-trisphosphate receptor (InsP(3)R1) is an intracellular calcium (Ca(2+)) release channel that plays an important role in neuronal function. In yeast two-hybrid screen of rat brain cDNA library with the InsP(3)R1 carboxy-terminal bait we isolated multiple clones of neuronal cytoskeletal protein 4.1N. We mapped the 4.1N-interaction site to a short fragment (50 amino acids) within the carboxy-terminal tail of the InsP(3)R1 and the InsP(3)R1-interaction site to the carboxy-terminal domain (CTD) of 4.1N. We established that InsP(3)R1 carboxy-terminal binds selectively to the CTDDelta alternatively spliced form of the 4.1N protein. In biochemical experiments we demonstrated that 4.1N and InsP(3)R1 specifically associate in vitro. We showed that both 4.1N and InsP(3)R1 were enriched in synaptic locations and immunoprecipitated the 4.1N-InsP(3)R1 complex from rat brain synaptosomes. In biochemical experiments we demonstrated a possibility of InsP(3)R1-4.1N-CASK-syndecan-2 quaternary complex formation. From our findings we hypothesize that InsP(3)R1-4.1N association may play a role in InsP(3)R1 localization or Ca(2+) signaling in neurons.


Population biology of the little gulper shark Centrophorus uyato in Lebanese waters.
A total of 38 individuals of the heavily exploited little gulper shark Centrophorus uyato were collected from Lebanese coastal waters using bottom longlines and trammel nets of different meshes at depths ranging from 115 to 600 m between May 2013 and February 2014. Their total lengths were between 45 and 94 cm and their total mass was from 870 to 6700 g. The sex ratio was not significantly different from 1:1, with 20 males and 18 females, but bathymetric sexual segregation occurred. Catch per net setting (CNS) was used as a relative abundance index to detect spatial distribution; trammel nets showed largest CNS ranging from 4·9 to 5·45 kg per unit effort in the north and south, at depths from 120 to 200 m, during spring and summer. The mass-length relationships demonstrated negative allometric growth (b < 3) (males: MT = 0.3585LT2·071 , r2 = 0·94; females: MT = 0.0239LT2·735 , r2 = 0·64). The condition factor as well as the gonado-somatic and hepato-somatic indices of C. uyato in the study area were also calculated. The results are discussed in relation to the distribution, growth and reproduction as well as the management of C. uyato.


Recovered LiCoO2 as anode materials for Ni/Co power batteries.
LiCoO(2) material is recovered from spent lithium-ion batteries and investigated as anode materials for Ni/Co power batteries for the first time. LiCoO(2) electrodes with a small amount of S-doping display excellent electrochemical properties. The electrochemical reactions occurring on M0 electrodes during the first several cycles and after being activated are proposed, respectively. A function mechanism of S powder on M10 electrode is also proposed.


Clinicopathological features of progressive renal involvement in TAFRO syndrome: A case report and literature review.
TAFRO syndrome is a systemic inflammatory disease characterized by a constellation of symptoms: Thrombocytopenia, Anasarca, MyeloFibrosis, Renal dysfunction, and Organomegaly. Progressive renal insufficiency is a predominant symptom; however, the mechanism of acute kidney injury (AKI) remains unclear, probably because severe thrombocytopenia prevents kidney biopsy. We report a rare case of TAFRO syndrome with histologically confirmed renal involvement. A 70-year-old man developed fever, anasarca, AKI, thrombocytopenia, and hepatosplenomegaly. Plasma vascular endothelial growth factor and serum interleukin-6 levels were significantly elevated. The diagnosis of TAFRO syndrome was made based on his clinical and laboratory findings. Kidney biopsy was performed for the evaluation of AKI and provided a diagnosis of membranoproliferative glomerulonephritis-like lesions due to endothelial injury. Glomerular capillary lumens were extremely narrowed or occluded by endothelial swelling, and marked widening of the subendothelial space by electron-lucent material resulted in mesangiolysis and a double-contoured glomerular basement membrane with no immune complex deposits. The patient required temporary hemodialysis due to oliguric AKI, but steroid therapy rapidly improved renal function. Typically, patients with progressive renal involvement in TAFRO syndrome rapidly develop oliguric or anuric AKI. This report suggests that the reduction of glomerular perfusion by glomerular endothelial injury might be a primary factor in the progressive AKI of TAFRO syndrome. Our case and the literature review indicate that steroid and/or biological therapies result in highly favorable renal outcomes in patients with progressive AKI in TAFRO syndrome.


Clinical trial design in the age of molecular profiling.
The accelerating science of molecular profiling has necessitated a rapid evolution in clinical trial design. Traditional clinical research begins with Phase I studies to characterize dose-limiting toxicities and defines maximally tolerated doses of drugs in small numbers of patients. Traditional Phase II studies test these drugs at the doses discovered during Phase I drug development in small numbers of patients evaluating efficacy and safety. Phase III studies test new therapies to demonstrate improved activity or improved tolerability compared with a standard of care regimen or a placebo. The rapid advances in the understanding of signal transduction, and the identification of new potential diagnostic and therapeutic targets, now require the design and implementation of molecular clinical trials that are very different than traditional Phase I, II, or III trials. The main differentiating factor is the use of a molecular end point to stratify a subset of patients to receive a specific treatment regimen. This chapter focuses on the issues surrounding (a) the definition of clinical end points and the assessment of tumor response; (b) clinical trial design models to define the targeted pathway; and (c) the need for appropriate biomarkers to monitor the response.


Stimulating spectrum of human recombinant multi-CSF (IL-3) on human marrow precursors: importance of accessory cells.
Recently, human multi-CSF was obtained by molecular cloning. In the present study, the effects of multi-CSF in vitro were investigated by comparative culture of whole bone marrow or progenitor cells obtained by sorting the cell fraction that binds the monoclonal antibody (MoAb) B13C5 (CD 34). Multi-CSF stimulated erythroid (BFU-E), multipotential (CFU-GEMM) and eosinophil (CFU-Eo) colonies in cultures of the progenitor cell enriched fraction, whereas (besides BFU-E, CFU-GEMM, and CFU-Eo) granulocyte (CFU-G), granulocyte-macrophage (CFU-GM), and macrophage (CFU-M) colony-forming cells also were stimulated by multi-CSF when unfractionated bone marrow was cultured. Reconstitution of the progenitor cell fraction (B13C5 positive) with the B13C5-negative population restored the broad spectrum of progenitor cell stimulation. This suggested that accessory cells are required for expression of the full spectrum of progenitor cell stimulation by multi-CSF. Subsequently, specific marrow cell populations, including T lymphocytes, granulocytic cells, and monocytes, were prepared by using selected MoAbs in complement-mediated lysis or cell sorting, added to cultures of hematopoietic progenitors and tested for accessory cell function. The results demonstrate that small numbers of monocytes permit the stimulation of CFU-G, CFU-GM, and CFU-M by multi-CSF. These monocyte-dependent stimulating effects on CFU-G, CFU-GM, and CFU-M could also be achieved by adding recombinant GM-CSF as a substitute for monocytes to the cultures. Therefore, multi-CSF most likely has direct stimulative effects on BFU-E, CFU-GEMM, and CFU-Eo and indirect effects on CFU-G, CFU-GM, and CFU-M in the presence of monocytes.


Superconducting Quantum Interference in Twisted van der Waals Heterostructures.
We demonstrate the formation of both Josephson junctions and superconducting quantum interference devices (SQUIDs) using a dry transfer technique to stack and deterministically misalign mechanically exfoliated flakes of NbSe2. The current-voltage characteristics of the resulting twisted NbSe2-NbSe2 junctions are found to be sensitive to the misalignment angle of the crystallographic axes, opening up a new control parameter for optimization of the device performance, which is not available in thin-film-deposited junctions. A single lithographic process has then been implemented to shape Josephson junctions into SQUID geometries with typical loop areas of ∼25 μm2 and weak links ∼600 nm wide. At T = 3.75 K in an applied magnetic field, these devices display large stable current and voltage modulation depths of up to ΔIc ∼ 75% and ΔV ∼ 1.4 mV, respectively.


Steroids and spinal cord injury: revisiting the NASCIS 2 and NASCIS 3 trials.
The National Acute Spinal Cord Injury Study (NASCIS) 2 and 3 trials are often cited as evidence that high-dose methylprednisolone is an efficacious intervention in the management of acute spinal cord injury. Neither of these studies convincingly demonstrate the benefit of steroids. There are concerns about the statistical analysis, randomization, and clinical end points. Even if the putative gains are statistically valid, the clinical benefits are questionable. Furthermore, the benefits of this intervention may not warrant the possible risks. This paper comments on these two clinical trials.


Antibody validation of immunohistochemistry for biomarker discovery: recommendations of a consortium of academic and pharmaceutical based histopathology researchers.
As biomarker discovery takes centre-stage, the role of immunohistochemistry within that process is increasing. At the same time, the number of antibodies being produced for "research use" continues to rise and it is important that antibodies to be used as biomarkers are validated for specificity and sensitivity before use. This guideline seeks to provide a stepwise approach for the validation of an antibody for immunohistochemical assays, reflecting the views of a consortium of academic and pharmaceutical based histopathology researchers. We propose that antibodies are placed into a tier system, level 1-3, based on evidence of their usage in immunohistochemistry, and that the degree of validation required is proportionate to their place on that tier.


Structural Studies Revealed Active Site Distortions of Human Furin by a Small Molecule Inhibitor.
Proprotein convertases (PCs) represent highly selective serine proteases that activate their substrates upon proteolytic cleavage. Their inhibition is a promising strategy for the treatment of several pathologies including cancer, atherosclerosis, hypercholesterolaemia, and infectious diseases. Here, we present the first experimental complex of furin with a non-substrate-like small molecule inhibitor, and the X-ray structure of the enzyme complexed to the small molecule inhibitor 1 at 1.9 Å resolution. Two molecules of inhibitor 1 were found to interact with furin. One is anchored at the S4 pocket of the enzyme and interferes directly with the conformation and function of the catalytic triade; the other molecule shows weaker binding and interacts with a distant, less conserved region of furin. The observed binding modes represent a new inhibition strategy of furin and imply the possibility to attain specificity among the PCs providing an innovative starting point of structure guided inhibitor development for furin.


The effect of gas cooking on bronchial hyperresponsiveness and the role of immunoglobulin E.
Some studies have shown an association between gas cooking and respiratory symptoms. This study investigated whether gas cooking affects bronchial responsiveness and whether particular subjects are more sensitive to this effect. Multiple linear regression analysis was performed with the dose-response slope (Percentage fall in forced expiratory volume in one second (FEV1) divided by total dose of methacholine given) as the dependent variable in 1,921 subjects from a random sample of the Dutch population, aged 20-70 yrs. Whether the association was different according to sex, age, total immunoglobulin (Ig)E, specific IgE to inhalant allergens or smoking habits was tested by including interaction terms into the regression model. Subjects who used gas for cooking had a higher prevalence of bronchial hyperresponsiveness (provocative dose causing a 20% fall in FEV1 (PD20) < or = 2 mg) than those who used electricity (21% versus 14%) and this was dependent on the presence of atopy. Especially subjects with total IgE levels in the highest quartile had a significantly higher dose-response slope when using gas for cooking. This was independent of the presence of specific IgE to inhalant allergens. These results show increased bronchial responsiveness with gas cooking, which was only found in subjects with high total immunoglobulin E levels. This suggests that atopic subjects are sensitive to adverse effects of gas cooking on respiratory health.


MIF inhibition interferes with the inflammatory and T cell-stimulatory capacity of NOD macrophages and delays autoimmune diabetes onset.
Macrophages contribute in the initiation and progression of insulitis during type 1 diabetes (T1D). However, the mechanisms governing their recruitment into the islets as well as the manner of retention and activation are incompletely understood. Here, we investigated a role for macrophage migration inhibitory factor (MIF) and its transmembrane receptor, CD74, in the progression of T1D. Our data indicated elevated MIF concentrations especially in long-standing T1D patients and mice. Additionally, NOD mice featured increased MIF gene expression and CD74+ leukocyte frequencies in the pancreas. We identified F4/80+ macrophages as the main immune cells in the pancreas expressing CD74 and showed that MIF antagonism of NOD macrophages prevented their activation-induced cytokine production. The physiological importance was highlighted by the fact that inhibition of MIF delayed the onset of autoimmune diabetes in two different diabetogenic T cell transfer models. Mechanistically, macrophages pre-conditioned with the MIF inhibitor featured a refractory capacity to trigger T cell activation by keeping them in a naïve state. This study underlines a possible role for MIF/CD74 signaling pathways in promoting macrophage-mediated inflammation in T1D. As therapies directed at the MIF/CD74 pathway are in clinical development, new opportunities may be proposed for arresting T1D progression.


[Possible participation of lactic acid in formation of a non-mediator agent stimulating the heart during exposure of the myocardium to sympathetic influences].
Release of A-factor, the physiologically active substance of a non--mediatory nature under sympathetic stimulation, depends upon the glycolytic process. To examine the assumption that A-factor may be a glycolytic product, lactic acid was studied in particular, and it was shown that A-factor is not identical to lactic acid. However it was found out that lactic acid participates in the release of A-factor in the heart. It was shown that under the effect of lactic acid a substance having high cardiostimulating activity was released in the frog heart muscle. This cardiostimulating substance as well as A-factor increases the amplitude of contractions of A-factor, was not abolished with beta-sympathicolytic inderal. The ability of both substances to stimulate heart activity was preserved after 20 min heating in a water bath. The cardiostimulating substance similar to A-factor increases heart sensitivity to sympathetic and decreases it to parasympathetic stimulations. It may be suggested that the cardiostimulating substance released in the heart under the effect of lactic acid is identical to A-factor released in the myocardium under the action of catecholamines and stimulation of the sympathetic nerve.


The Functional Role of SEC23 in Vesicle Transportation, Autophagy and Cancer.
SEC23, the core component of the coat protein complex II (COPII), functions to transport newly synthesized proteins and lipids from the endoplasmic reticulum (ER) to the Golgi apparatus in cells for secretion. SEC23 protein has two isoforms (SEC23A and SEC23B) and their aberrant expression and mutations were reported to cause human diseases and oncogenesis, whereas SEC23A and SEC23B may have the opposite activity in human cancer, for a reason that remains unclear. This review summarizes recent research in SEC23, COPII-vesicle transportation, autophagy, and cancer.


Polysubstance use by sexual identity among US adults, 2021.
Understanding polysubstance use among US sexual minority adults is important to serve as a population-level baseline to promote health equity around substance use prevention and public health strategies. This study quantifies the number of substances used by sexual identity among US adults. We used the 2021 National Survey on Drug Use and Health and included adults (aged ≥18) (unweighted n = 47,291). We conducted multivariable Poisson regression models to examine the number of substances used in the past year (count variable; range: 0-18) by sexual identity ("heterosexual", "gay/lesbian", "bisexual", "unknown" [don't know, refused, blank]), after adjusting for covariates (i.e., age, sex, race/ethnicity, income level, education level, having insurance status, living in urban area, past-year distress level (Kessler-6), any drug or alcohol use disorder in the past year, and sexual attraction). Of the total sample (51.4% were female, 12.1% were non-Hispanic Black and 17.0% were Hispanic adults), 88.3% were heterosexual, 2.4% were gay/lesbian, 5.0% were bisexual adults, and 4.3% reported "unknown" sexual identity. After adjusting for covariates, a greater number of substances were used in the past year among gay/lesbian individuals (aIRR = 1.44, 95% CI = 1.09, 1.75), bisexual individuals (aIRR =1.34, 95% CI = 1.26, 1.41), and individuals reporting an "unknown" sexual identity (aIRR = 1.22, 95% CI = 1.09, 1.36) (vs. heterosexual adults). Tailored substance use prevention and public health strategies specializing in sexual minority populations are warranted.


Identification of 1-({[1-(4-fluorophenyl)-5-(2-methoxyphenyl)-1H-pyrazol-3-yl]carbonyl}amino)cyclohexane carboxylic acid as a selective nonpeptide neurotensin receptor type 2 compound.
Compounds active at neurotensin receptors (NTS1 and NTS2) exert analgesic effects on different types of nociceptive modalities, including thermal, mechanical, and chemical stimuli. The NTS2 preferring peptide JMV-431 (2) and the NTS2 selective nonpeptide compound levocabastine (6) have been shown to be effective in relieving the pain associated with peripheral neuropathies. With the aim of identifying novel nonpeptide compounds selective for NTS2, we examined analogues of SR48692 (5a) using a FLIPR calcium assay in CHO cells stably expressing rat NTS2. This led to the discovery of the NTS2 selective nonpeptide compound 1-({[1-(4-fluorophenyl)-5-(2-methoxyphenyl)-1H-pyrazol-3-yl]carbonyl}amino)cyclohexane carboxylic acid (NTRC-739, 7b) starting from the nonselective compound 5a.


Nutrition in Ultra-Endurance: State of the Art.
Athletes competing in ultra-endurance sports should manage nutritional issues, especially with regards to energy and fluid balance. An ultra-endurance race, considered a duration of at least 6 h, might induce the energy balance (i.e., energy deficit) in levels that could reach up to ~7000 kcal per day. Such a negative energy balance is a major health and performance concern as it leads to a decrease of both fat and skeletal muscle mass in events such as 24-h swimming, 6-day cycling or 17-day running. Sport anemia caused by heavy exercise and gastrointestinal discomfort, under hot or cold environmental conditions also needs to be considered as a major factor for health and performance in ultra-endurance sports. In addition, fluid losses from sweat can reach up to 2 L/h due to increased metabolic work during prolonged exercise and exercise under hot environments that might result in hypohydration. Athletes are at an increased risk for exercise-associated hyponatremia (EAH) and limb swelling when intake of fluids is greater than the volume lost. Optimal pre-race nutritional strategies should aim to increase fat utilization during exercise, and the consumption of fat-rich foods may be considered during the race, as well as carbohydrates, electrolytes, and fluid. Moreover, to reduce the risk of EAH, fluid intake should include sodium in the amounts of 10⁻25 mmol to reduce the risk of EAH and should be limited to 300⁻600 mL per hour of the race.


Influence of High Pass Filter Settings on Motor Evoked Potentials.
Motor evoked potentials (MEP), obtained by transcranial magnetic stimulation (TMS) are a common tool in clinical research and diagnostic. Nevertheless, reports regarding the influence of filter settings on MEP are sparse. Here, we compared MEP amplitudes and signal to noise ratio (SNR) using multiple high pass filter (HPF) and notch filter settings. Twenty healthy subjects were enrolled in the study. Recruitment curves were obtained with HPF settings varied at 10, 20, 50, and 100 Hz. The four HPF settings were tested both with and without 50 Hz active notch filter. Low pass filter was kept constant at 5 kHz. MEP amplitudes with HPF at 10 and 20 Hz were significantly higher than at 100 Hz, regardless of the notch filter. However, SNR did not differ among HPF settings. An active notch filter significantly improved SNR. The reduction in MEP amplitudes with HPF above 20 Hz may be due to noise reduction, since the different HPF conditions did not alter SNR. Thus, higher HPF above 50 Hz may be an option to reduce noise, the use of a notch filter may even improve SNR. Our findings are relevant for the selection of filter settings and might be of importance to any researcher who utilizes TMS-MEP.


Screening for cancers with a good prognosis: The case of testicular germ cell cancer.
To determine, using testicular germ cell cancer screening as an example, whether screening can also be effective for cancers with a good prognosis. Based on the Dutch incidence, stage distribution, and survival and mortality data of testicular germ cell cancer, we developed a microsimulation model. This model simulates screening scenarios varying in screening age, interval, self-examination or screening by the general practitioner (GP), and screening of a defined high-risk group (cryptorchidism). For each scenario, the number of clinically and screen-detected cancers by stage, referrals, testicular germ cell cancer deaths, and life-years gained were projected. Annual self-examination from age 20 to 30 years resulted in 767 cancers detected per 100,000 men followed over life-time, of which 123 (16%) by screening. In this scenario, 19.2 men died from the disease, 4.7 (20%) less than without screening, and 230 life-years were gained. Around 14,000 visits to the GP and 2080 visits to an urologist were required. This scenario resulted in the most favorable ratio between extra visits to the GP or urologist and deaths prevented (1418 and 116 respectively). Monthly screening, or screening until age 40 resulted in less favorable ratios. Self-examination by only the high-risk population prevented 1.0 death per 100,00 men in the general population. In all scenarios, 46-50 life-years were gained for each testicular germ cell cancer death prevented. Despite the good prognosis, self-examination at young ages for testicular germ cell cancer could be considered.


LC Data QUEST: A Technical Architecture for Community Federated Clinical Data Sharing.
The University of Washington Institute of Translational Health Sciences is engaged in a project, LC Data QUEST, building data sharing capacity in primary care practices serving rural and tribal populations in the Washington, Wyoming, Alaska, Montana, Idaho region to build research infrastructure. We report on the iterative process of developing the technical architecture for semantically aligning electronic health data in primary care settings across our pilot sites and tools that will facilitate linkages between the research and practice communities. Our architecture emphasizes sustainable technical solutions for addressing data extraction, alignment, quality, and metadata management. The architecture provides immediate benefits to participating partners via a clinical decision support tool and data querying functionality to support local quality improvement efforts. The FInDiT tool catalogues type, quantity, and quality of the data that are available across the LC Data QUEST data sharing architecture. These tools facilitate the bi-directional process of translational research.


Neonatal mortality among preterm infants admitted to neonatal intensive care units in India and Pakistan: A prospective study.
To explore potential reasons for differences in preterm neonatal mortality in neonatal intensive care units (NICUs) in India and Pakistan. A prospective observational study, the Project to Understand and Research Stillbirth and Preterms in Southeast Asia (PURPOSe) was conducted July 2018 to February 2020. Three hospitals in Davangere, India, and a large public hospital in Karachi, Pakistan. Of a total of 3,202 preterm infants enrolled, 1,512 were admitted to a study NICU. We collected data for neonates, including length of stay, diagnoses, and diagnostic tests. Neonatal mortality, tests performed, diagnoses ascertained. For infants of equivalent weights and gestational ages, neonatal mortality in Pakistan was twice that in the Indian NICU. The mean newborn length of stay in Pakistan was 2 days compared with 10 days for India. Fewer diagnostics and other investigations were used to determine neonatal condition or guide treatment in the Pakistani NICU. Because of limited information from testing in Pakistan concerning clinical respiratory distress, respiratory distress syndrome appeared to be over-diagnosed, whereas other conditions including pneumonia, sepsis, necrotising entercolitis and intraventricular haemorrhage were rarely diagnosed. In the Pakistani site, the limited resources available to the NICU appeared related to a shorter length of stay and decreased diagnostic testing, likely explaining the higher mortality. With improved care, reduction in mortality among preterm neonates should be achievable.


Cultured human trabecular meshwork cells express aquaporin-1 water channels.
The identification and characterization of aquaporin-1 water channels and other related proteins has provided a molecular explanation for the enhanced permeability of a variety of epithelial tissues. Previously, we documented the distribution of aquaporin-1 in the human eye, which included the trabecular meshwork; the primary outflow channel for aqueous humor. The goal of this study was to determine if aquaporin-1 could be detected in cultures of human trabecular meshwork cells. Using primers specific for aquaporin-1, reverse transcription combined with polymerase chain reaction yielded a product of the appropriate size with total RNA prepared from the human trabecular meshwork cells. The presence of this product and its size (298 base pairs), is consistent with the presence of an aquaporin-1 message in these cells. Indirect immunofluorescence microscopy with affinity purified antibodies against a fusion protein containing the carboxy tail of aquaporin-1 showed specific labeling of the plasma membrane and immunoblotting identified a band of Mr 28,000 which agrees with the molecular size of aquaporin-1. The presence of aquaporin-1 in human trabecular meshwork cells, the predominant cell-type of the primary outflow region of the human eye, suggests that water channels may be involved with the movement of aqueous fluid out of the eye. In addition, the existence of aquaporin-1 on cultures of human trabecular meshwork cells provides an in vitro model to study the endogenous expression of aquaporin-1 and its possible role in the regulation of aqueous outflow.


Establishing Age- and Sex-Specific Norms for Pediatric Return-to-Sports Physical Performance Testing.
Graft tears and contralateral anterior cruciate ligament (ACL) tears are common in pediatric athletes after ACL reconstruction. Use of objective return-to-sports (RTS) criteria, in particular physical performance tests (PPTs), is believed to reduce the incidence of secondary injury; however, pediatric norms for these tests are unknown. To establish a proof of concept for the creation of age- and sex-based norms for commonly used RTS PPTs in healthy pediatric athletes, allowing the creation of growth curves for clinical referencing. Cross-sectional study; Level of evidence, 3. A total of 100 healthy people who were between the ages of 6 and 18 years and involved in organized sports were enrolled, with even distributions of age and sex. All participants underwent 9 common RTS PPTs: stork test, stork test on Bosu, single-leg squat, single-leg squat on Bosu, clockwise and counterclockwise quadrant hops, single-leg hop for distance, 6-m timed hop, and triple crossover hop for distance. Mean performance across limbs was calculated for each individual. Chronological age, height, weight, sex, and self-reported Pubertal Maturational Observational Scale (PMOS) score were recorded. Univariable and multivariable models were created for each PPT, assessing the importance of the recorded descriptive variables. Quantile regression was used to create growth curves for each PPT. The cohort was 52% female, and the mean ± standard deviation age was 11.7 ± 3.6 years. PMOS was highly correlated with age (r = 0.86) and was excluded from the regressions. In univariable regression, age, height, and weight were strong predictors of performance for all PPTs, whereas sex was a predictor of performance on the single-leg and triple crossover hops for distance (with males outperforming females). Height and weight were excluded from multivariable regression because of multicollinearity with age. Multivariable regression showed predictive patterns for age and sex that were identical to those shown in the univariable analysis. Given ceiling effects, quantile regression for the stork tests was not possible, but quantile regression growth curves were successfully created for the 7 remaining PPTs. Chronological age and sex accurately predicted performance on common RTS PPTs in pediatric patients. The growth curves presented herein could assist clinicians with benchmarking pediatric patients postoperatively against a healthy athletic cohort.


Multiple Sessions of High-Frequency Repetitive Transcranial Magnetic Stimulation as a Potential Treatment for Gambling Addiction: A 3-Month, Feasibility Study.
Gambling disorder (GD) is a behavioral addiction, in which dysfunctions in prefrontal activity have been proposed as relevant pathophysiological correlates. The aim of the present study was to preliminarily investigate the feasibility of a noninvasive neuromodulation intervention targeting the prefrontal cortex to treat GD in an open-label setting. We included 8 treatment-seeking patients with GD (7 males; 1 female; mean age: 40.6 ± 11.2). The study consisted of 3 phases: (1) outpatient screening phase, (2) 2-week intensive repetitive transcranial magnetic stimulation (rTMS) treatment phase (twice daily, 5 days/week for 2 weeks); and (3) 3-month maintenance follow-up phase (twice daily, once a week). Each high-frequency (15 Hz) rTMS session was delivered targeting the left dorsolateral prefrontal cortex. GD severity and treatment response were assessed at the baseline and during the follow-up. No relevant side effect was reported. We found a 71.2% Gambling Symptom Assessment Scale mean score reduction after 2 weeks of rTMS treatment; the days spent gambling decreased from 19.63 ± 7.96 to 0.13 ± 0.35 days. Clinical improvements were maintained throughout the study period. The lack of a control group limits the interpretation of these results. In conclusion, these results consolidate the rationale that rTMS interventions deserve further investigation as a potential treatment for GD. These protocols should be tested in larger randomized controlled studies, to determine the real benefits of neuromodulation in the clinical course of patients with GD. Registration Number: ClinicalTrials.gov Identifier NCT03336879.


Electrocardiographical case. Young woman with frequent syncope attacks.
A 29-year-old woman with frequent syncope attacks was referred for electrophysiological study and consideration for radio-frequency ablation of her ventricular arrhythmias. Her ECG showed features of right ventricular outflow tract premature contraction. Differential diagnoses for the causes of syncope in this patient include: right ventricular outflow tract tachycardia, arrythmogenic right ventricular dysplasia, and neurocardiogenic syncope. She underwent a tilt table test, which showed a malignant cardioinhibitory response. She developed abrupt syncope with 32 seconds of asystole during the test. She was given intravenous atropine and was resuscitated. A dual chamber rate-responsive pacemaker was implanted for her the next day. She was discharged well subsequently. Although the prognosis in patients with prolonged aystole in malignant vasovagal syncope is unknown, most doctors will still choose to implant a permanent pacemaker for patients with malignant neurocardiogenic syncope when the sinus arrest is prolonged.


Supersaturation and Solubilization upon In Vitro Digestion of Fenofibrate Type I Lipid Formulations: Effect of Droplet Size, Surfactant Concentration and Lipid Type.
Lipid-based formulations (LBF) enhance oral drug absorption by promoting drug solubilization and supersaturation. The aim of the study was to determine the effect of the lipid carrier type, drop size and surfactant concentration on the rate of fenofibrate release in a bicarbonate-based in vitro digestion model. The effect of the lipid carrier was studied by preparing type I LBF with drop size ≈ 2 µm, based on medium-chain triglycerides (MCT), sunflower oil (SFO), coconut oil (CNO) and cocoa butter (CB). The drop size and surfactant concentration effects were assessed by studying MCT and SFO-based formulations with a drop size between 400 nm and 14 µm and surfactant concentrations of 1 or 10%. A filtration through a 200 nm filter followed by HPLC analysis was used to determine the aqueous fenofibrate, whereas lipid digestion was followed by gas chromatography. Shorter-chain triglycerides were key in promoting a faster drug release. The fenofibrate release from long-chain triglyceride formulations (SFO, CNO and CB) was governed by solubilization and was enhanced at a smaller droplet size and higher surfactant concentration. In contrast, supersaturation was observed after the digestion of MCT emulsions. In this case, a smaller drop size and higher surfactant had negative effects: lower peak fenofibrate concentrations and a faster onset of precipitation were observed. The study provides new mechanistic insights on drug solubilization and supersaturation after LBF digestion, and may support the development of new in silico prediction models.


Room-temperature Cu(II)-catalyzed chemo- and regioselective ortho-nitration of arenes via C-H functionalization.
An efficient Cu-catalyzed chemo- and regioselective ortho-nitration of N,1-diaryl-5-aminotetrazoles and N,4-diaryl-3-amino-1,2,4-triazoles has been described with good functional group compatibility. The procedure features the use of an operationally simple protocol utilizing the commercially available less toxic CuCl2·2H2O as catalyst and Fe(NO3)3·9H2O as nitration source at room temperature. Removal of the 5-aminotetrazole directing group has been demonstrated using base hydrolysis to afford substituted 2-nitroanilines.


Implications of myocardial transformation for cardiac energetics.
The influence of isoenzyme pattern of myosin on cardiac energetics was investigated in a modified in situ heart-lung preparation in the rat. Chronic pressure load (spontaneous hypertension, aortic stenosis, Goldblatt hypertension), intermittent feeding, and swim-training elicited redistribution in the concentration of alpha chains of myosin ranging from 18 to 94%. The influence of isoenzyme pattern of myosin on cardiac energetics could be quantitatively assessed by extrapolation of the regression line of oxygen and substrate consumption related to tension time index. Fast myocardium with 100% alpha chains had an ATP and oxygen consumption which exceeded that of slow myocardium with 0% alpha chains by about 60%. This corresponds well to the state of activity of myofibrillar ATPase of fast myocardium which also exceeds that of slow myocardium by about 50%. Furthermore it could be shown that acute increase in the ATPase activity depends on the isoenzyme pattern of myosin. Under the influence of catecholamines the oxygen consumption related to tension time index increased by 30-40% in fast myocardium, whereby in a myocardium with 40% alpha chains no increase in oxygen consumption per unit tension time index was observed, when catecholamines were applied.


The Fixion nail in the lower limb. Preliminary results.
The Fixion nail is an intramedullary nail made of stainless steel, that may be used to treat fractures of the humerus and in the lower limb. This nail is made up of a cylinder that has 4 longitudinal bars which, once inserted in the bone segment, expands via a pressure hydraulic system using physiological solution. The nail may be introduced without reaming and after its expansion it adheres to the bone walls producing stabilization of the fracture. In order to guarantee good stability, the apex of the nail must exceed the fracture line by 5 cm. Indications were initially limited to type 32 and 42 A1-2-3 fractures of the mid third of the lower limb, to then be expanded to type B1 fractures in the AO system. A total of 20 nails were used in 19 patients, with an average time of 7 days for partial weight-bearing and 40 days for total weight-bearing, achieving consolidation on day 18 after an average of 5 months in the femur and 4 months in the tibia. Reaming of the intramedullary canal was carried out in all of the cases.


Laryngeal mask airway versus bag-mask ventilation or endotracheal intubation for neonatal resuscitation.
Providing effective positive pressure ventilation is the single most important component of successful neonatal resuscitation. Ventilation is frequently initiated with a manual resuscitation bag and face-mask (BMV) followed by endotracheal intubation (ETT) if depression continues. These techniques may be difficult to perform successfully resulting in prolonged resuscitation or severe neonatal depression. The laryngeal mask airway (LMA) may achieve initial ventilation and successful resuscitation faster than a bag-mask device or endotracheal intubation. Among newborns requiring positive pressure ventilation for resuscitation, is effective ventilation and successful resuscitation achieved faster with the LMA compared with either BMV or ETT? The Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 3, 2004), MEDLINE (1966-November 2004), Pre-MEDLINE (November 15, 2004), CINAHL 1982-November 2004), reference lists of published trials, and Society for Pediatric Research abstracts were searched. Experts were contacted for additional references. Randomised and quasi-randomised trials Two reviewers independently evaluated studies, assessed methodologic quality, and extracted data using the Cochrane Neonatal Review Group criteria. Categorical treatment effects were described as relative risks and risk differences and continuous treatment effects were described as the mean difference. There were insufficient data to perform pooled analyses. No eligible studies compared the LMA with BMV. One small randomised controlled trial comparing the LMA with ETT when BMV had been unsuccessful was included. There was no statistically significant difference between the LMA and ETT with the exception of a clinically insignificant difference in time to complete insertion of the device favouring the ETT. The LMA can achieve effective ventilation during neonatal resuscitation in a time-frame consistent with current guidelines. There is no evidence to evaluate the relative efficacy and safety of the LMA compared with BMV as the primary airway device. A single, small randomised controlled trial found no clinically significant difference between the LMA and ETT when BMV was unsuccessful. Case series and case reports suggest that the LMA can provide an effective rescue airway during resuscitation if both BMV and ETT have been unsuccessful. A well-designed randomised controlled trial comparing the LMA with BMV during neonatal resuscitation is warranted.


Cardiogenic shock following administration of propofol and fentanyl in a healthy woman: a case report.
Cardiogenic shock is very uncommon in healthy people. The differential diagnosis for patients with acute heart failure in previously healthy hearts includes acute myocardial infarction and myocarditis. However, many drugs can also depress myocardial function. Propofol and fentanyl are frequently used during different medical procedures. The cardiovascular depressive effect of both drugs has been well established, but the development of cardiogenic shock is very rare when these agents are used. After a minor surgical intervention, a 32-year-old Caucasian woman with no significant medical history went into sudden hemodynamic deterioration due to acute heart failure. An urgent echocardiogram showed severe biventricular dysfunction and an estimated left ventricular ejection fraction of 20%. Extracorporeal life support and mechanical ventilation were required. Five days later her ventricular function had fully recovered, which allowed the progressive withdrawal of medical treatment. Prior to her hospital discharge, cardiac MRI showed neither edema nor pathological deposits on the delayed contrast enhancement sequences. At her six-month follow-up examination, the patient was asymptomatic and did not require treatment. Although there are many causes of cardiogenic shock, the presence of abrupt hemodynamic deterioration and the absence of a clear cause could be related to the use of propofol and fentanyl.


Hepatic oxygen supply during early and late sepsis in the rat.
Hepatic oxygen supply and related parameters were evaluated during early (8-10 h) and late (18-26 h) stages of lethal septic peritonitis induced in rats by cecal ligation and puncture. In the early phase, hepatic pO2 was reduced significantly (mean 9.3 versus 22.2 mmHg in sham-operated controls). This was accompanied by elevations of serum glutamic oxaloacetic transaminase and glutamic pyruvic transaminase (p less than 0.02 and p less than 0.10, respectively), and increased hematocrit (p less than 0.001). Total body oxygen consumption was decreased by 19% (p less than 0.001). By the late stage of sepsis, hepatic oxygen supply was depressed profoundly (mean 2.0 versus 24.5 mmHg in controls) concomitant with further elevations of serum transaminases. Lactacidemia, respiratory alkalosis, and tachycardia were also present. Hypoxemia and hypotension were not observed during either early or late stages. Our results may be interpreted as representing early and progressive impairment of hepatic oxygen delivery in rats subjected to cecal ligation and puncture.


Hand Mirror Cells and Hypercalcemia: A Rare Presentation of Pediatric Acute Lymphoblastic Leukemia.
Hand mirror cell (HMC) leukemia is a variant of acute lymphoblastic leukemia previously described in the adult population where lymphoblasts manifest distinctive hand mirror morphologic features. HMC has been previously identified in 23% of childhood acute lymphoblastic leukemia patients, but its prognostic significance in children is not well understood. Hypercalcemia is also uncommon in childhood leukemias. Hypercalcemia associated with HMC leukemia has not been previously reported. We report a 5-year-old boy with HMC B-lymphoblastic leukemia who presented with hypercalcemia.


Interventions to prevent obesity in 0-5 year olds: an updated systematic review of the literature.
The small number and recency of the early childhood obesity-prevention literature identified in a previous review of interventions to prevent obesity, promote healthy eating, physical activity, and/or reduce sedentary behaviors in 0-5 year olds suggests this is a new and developing research area. The current review was conducted to provide an update of the rapidly emerging evidence in this area and to assess the quality of studies reported. Ten electronic databases were searched to identify literature published from January 1995 to August 2008. interventions reporting child anthropometric, diet, physical activity, or sedentary behavior outcomes and focusing on children aged 0-5 years of age. focusing on breastfeeding, eating disorders, obesity treatment, malnutrition, or school-based interventions. Two reviewers independently extracted data and assessed study quality. Twenty-three studies met all criteria. Most were conducted in preschool/childcare (n = 9) or home settings (n = 8). Approximately half targeted socioeconomically disadvantaged children (n = 12) and three quarters were published from 2003 onward (n = 17). The interventions varied widely although most were multifaceted in their approach. While study design and quality varied most studies reported their interventions were feasible and acceptable, although impact on behaviors that contribute to obesity were not achieved by all. Early childhood obesity-prevention interventions represent a rapidly growing research area. Current evidence suggests that behaviors that contribute to obesity can be positively impacted in a range of settings and provides important insights into the most effective strategies for promoting healthy weight from early childhood.


Age related microsatellite instability in T cells from healthy individuals.
Many immune functions decline with age and may jeopardize the elderly, as illustrated, for example by the significantly higher mortality rate from influenza in old age. Although innate and humoral immunity are affected by aging, it is the T cell compartment, which manifests most alterations. The mechanisms behind these alterations are still unclear, and several explanations have been offered including thymic involution and Telomere attrition leading to cell senescence. Age related accumulation of mutations has been documented and could serve as an additional mechanism of T cell dysfunction. One effective repair mechanism capable of rectifying errors in DNA replications is the mismatch repair (MMR) system. We previously reported a comparative examination of individual DNA samples from blood cells obtained at 10 year intervals from young and old subjects. We showed significantly higher rates of microsatellite instability (MSI), an indicator of MMR dysfunction in older subjects, compared to young. In the present study we confirm this result, using direct automated sequencing and in addition, we demonstrate that as CD8 lymphocytes from aged individuals, undergo repeated population doublings (PDs) in culture, they develop MSI. CD4 clones that also undergo repeated PDs in culture develop significant MSI as well. Elucidation of this previously unexplored facet of lymphocyte dynamics in relation to aging may help identify novel mechanisms of immunosenescence and pathways that could serve as targets for interventions to restore immune function.


Polymeric micelle nanoparticles for photodynamic treatment of head and neck cancer cells.
To encapsulate 5,10,15,20-tetrakis(meso-hydroxyphenyl)porphyrin (mTHPP), a photosensitizer, into polymeric micelles; characterize the micelles; and test in vitro photodynamic therapy efficacy against human head and neck cancer cells. A nanoparticle design, fabrication, and in vitro testing study. Polymer chemistry laboratory. Micelles encapsulating mTHPP were produced, and micellar size was measured. Ultraviolet visible spectra and fluorescence spectroscopy were used to characterize the mTHPP-loaded micelles. In vitro cell culture using HSC-3 and HN-5 cancer cells was performed to test the photodynamic therapy efficacy of the micelles using confocal microscopy and method of transcriptional and translational (MTT) assay. mTHPP was encapsulated with high loading efficiency (> 85%) and density (up to 17%) into micelles. Micelle size was 30.6 +/- 3.3 nm by transmission electron microscopy and 30.8 +/- 0.6 nm by dynamic light scattering. The absorption maximum for each sample was 418 nm, and fluorescent spectroscopy revealed quenching with maximal fluorescence at five percent loading. Significant cytotoxicity was observed with confocal microscopy when HSC-3 cells were treated with 10 percent mTHPP micelles, with 100 percent cytotoxicity within the zone of laser light exposure at 420 nm. Phototoxicity and dark toxicity against HSC-3 and HN-5 cells measured using the MTT assay with five and 10 percent loaded mTHPP micelles demonstrated greater than 90 percent cytotoxicity with photodynamic therapy and less than 10 percent dark toxicity at a micelle concentration of 25 microg/mL for both cell lines. Micelles were able to encapsulate and solubilize mTHPP at high loading densities with uniform size distribution. These micelles exhibit fluorescence and photodynamic therapy mediated cytotoxicity against head and neck cancer cells in vitro.


The impact of the COVID-19 pandemic on seafarers' mental health and chronic fatigue: Beneficial effects of onboard peer support, external support and Internet access.
While commercial shipping remained vital for maintaining global supply chains during the COVID-19 pandemic, measures imposed to control the spread of infection have disrupted crew changes and impacted interactions with port personnel and among crew members on board. Initial reports indicate that this affected work and life on board, the length of seafarers' time on board as well as seafarers' employment and family concerns. However, the consequences for seafarers' well-being are not well understood. The purpose of this study was to examine the effects of the COVID-19 pandemic on seafarers' mental health and chronic fatigue, and to analyze the role of potential mitigating factors, notably onboard peer support, external support and Internet quality. Survey responses from 622 seafarers on international commercial vessels were analyzed using structural equation modeling. Findings suggested that the impact of the pandemic increased seafarers' fatigue and mental health problems. However, they also indicated ways of mitigating the negative impact of the pandemic and increasing resilience by enhancing support from fellow crew members on board, ensuring the availability of external support and providing fast and reliable Internet access.


Effect of eriodictyol on the development of atopic dermatitis-like lesions in ICR mice.
Atopic dermatitis (AD) is a chronic, allergic, and inflammatory skin disease associated with eczema and dermatitis symptoms. Our previous studies have reported that eriodictyol extract inhibits immunoglobulin E (IgE)/Ag-induced type I hypersensitivity by suppressing the activation of proinflammatory cytokines, such as interleukin-4 (IL-4), and the expression of ceramide kinase. In this study, we investigated the inhibitory effect of eriodictyol on 2,4-dinitrochlorobenzene (DNCB)-induced AD-like skin lesions in ICR mice. Treatment with 2 mg/mL eriodictyol for DNCB-induced AD-like skin lesions in ICR mice improved scratching behavior and skin severity score. Histological analysis demonstrated that thickening of the skin lesions were significantly reduced in the eriodictyol-treated group. Also, eriodictyol suppressed the DNCB-mediated elevation of IgE serum levels. These results suggest that eriodictyol may be a potential therapeutic resource for AD and an adjunctive agent to control itchiness in AD.


Rotigotine transdermal patch for the treatment of Parkinson's disease and restless legs syndrome.
The nonergoline dopamine agonist rotigotine, is delivered transdermally using a silicone-based patch (Neupro(R); UCB Pharma GmbH), which promotes unidirectional drug flow from the transdermal system to the skin. Pharmacokinetic data show stable steady-state plasma concentrations over 24 h, maintained with once-daily patch administration. Stable plasma concentrations are reflected by stable concentrations in the brain, as has been shown in animal studies. This suggests a continuous stimulation of dopaminergic receptors, which may result in a reduction in or prevention of abnormal involuntary movements in Parkinson's disease (PD) after prolonged treatment. Clinical trials have demonstrated that rotigotine is efficacious as monotherapy for PD and restless legs syndrome (RLS), and open-label extension studies have shown its long-term efficacy. Furthermore, rotigotine can be used effectively in coadministration with levodopa, enabling a reduction of the levodopa treatment doses in PD. Transdermal application also yields favorable pharmacokinetics for rotigotine: rapid metabolism and lack of skin accumulation allow for good control of chronic administration or withdrawal by patch removal, and the transdermal application approach circumvents problems of gastrointestinal absorption and enables administration prior to, during or following surgery. In addition, no dose adaptation is required regarding gender or ethnicity, or for patients with impaired liver or kidney function or on hemodialysis. The safety profile of rotigotine transdermal patch is favorable; common side effects attributed to transdermal delivery or dopaminergic stimulation are generally mild to moderate in intensity. Importantly, augmentation of RLS is uncommon under long-term rotigotine treatment and dyskinesia in PD patients mostly developed only after levodopa initiation. Overall, rotigotine transdermal patch has demonstrated favorable clinical efficacy and tolerability in the treatment of PD and RLS.


Oncolytic potency and reduced virus tumor-specificity in oncolytic virotherapy. A mathematical modelling approach.
In the present paper, we address by means of mathematical modeling the following main question: How can oncolytic virus infection of some normal cells in the vicinity of tumor cells enhance oncolytic virotherapy? We formulate a mathematical model describing the interactions between the oncolytic virus, the tumor cells, the normal cells, and the antitumoral and antiviral immune responses. The model consists of a system of delay differential equations with one (discrete) delay. We derive the model's basic reproductive number within tumor and normal cell populations and use their ratio as a metric for virus tumor-specificity. Numerical simulations are performed for different values of the basic reproduction numbers and their ratios to investigate potential trade-offs between tumor reduction and normal cells losses. A fundamental feature unravelled by the model simulations is its great sensitivity to parameters that account for most variation in the early or late stages of oncolytic virotherapy. From a clinical point of view, our findings indicate that designing an oncolytic virus that is not 100% tumor-specific can increase virus particles, which in turn, can further infect tumor cells. Moreover, our findings indicate that when infected tissues can be regenerated, oncolytic viral infection of normal cells could improve cancer treatment.


Apodization and the point-spread autocorrelation function.
A novel function, the point-spread autocorrelation function (PSAF), which is closely related to the point-spread function, of a high numerical aperture microscope objective is introduced. The function is both experimentally measured and theoretically modeled for various apodization conditions. These include varying the effective numerical aperture of the objective, applying annuli of different size, and illuminating the objective with a spatially nonuniform intensity distribution. An excellent agreement between experimental data and theoretical modeling is obtained without the use of any fitting parameters. The PSAF technique is sensitive to the various apodization conditions, affecting both the width of the PSAF signal and the amplitude of the sidelobes. A potential use of the technique is the measurement of the effective numerical aperture and the optimization of the illumination conditions in complex microscopical systems.


The V3 loop-mimicking pseudopeptide 5[Kpsi(CH2N)PR]-TASP inhibits HIV infection in primary macrophage cultures.
The V3 loop-mimicking pseudopeptide 5[Kpsi(CH2N)PR]-TASP [psi(CH2N) representing a reduced peptide bond], which presents pentavalently the tripeptide Kpsi(CH2N)PR, is a potent inhibitor of HIV entry. By its capacity to bind specifically protein components on the cell surface, 5[Kpsi(CH2N)PR]-TASP blocks the attachment of virus particles to permissive CD4+ cells. Here, the inhibitory effect of 5[Kpsi(CH2N)PR]-TASP was investigated in monocyte-derived macrophages (MDMs) infected by the monocytotropic HIV-1(Ba-L) isolate. We show that 5[Kpsi(CH2N)PR]-TASP inhibits HIV-1(Ba-L) infection in a dose-dependent manner, with more than 90% inhibition at 2 microM concentration. On the other hand, the control 5[QPQ]-TASP construct and the monovalent Kpsi(CH2N)PR tripeptide have no effect even at high concentrations. Under such experimental conditions, the biotin-labeled 5[Kpsi(CH2N)PR]-TASP, but not the Kpsi(CH2N)PR construct, binds specifically to the surface of MDMs and forms a stable complex with the cell surface-expressed nucleolin, as has been demonstrated to be the case in peripheral blood mononuclear cells. Infection of MDMs by HIV-1(Ba-L) could also be inhibited by beta-chemokines RANTES and MIP-1beta. Interestingly, association of low concentrations of 5[Kpsi(CH2N)PR]-TASP and beta-chemokines results in a synergistic inhibitory effect on HIV infection compared with the effect observed with each reagent alone. The inhibitory effect of 5[Kpsi(CH2N)PR]-TASP in primary macrophage cultures point out its potential as an anti-HIV drug in cells, which are the natural viral targets.


Silicon-carbon bond inversions driven by 60-keV electrons in graphene.
We demonstrate that 60-keV electron irradiation drives the diffusion of threefold-coordinated Si dopants in graphene by one lattice site at a time. First principles simulations reveal that each step is caused by an electron impact on a C atom next to the dopant. Although the atomic motion happens below our experimental time resolution, stochastic analysis of 38 such lattice jumps reveals a probability for their occurrence in a good agreement with the simulations. Conversions from three- to fourfold coordinated dopant structures and the subsequent reverse process are significantly less likely than the direct bond inversion. Our results thus provide a model of nondestructive and atomically precise structural modification and detection for two-dimensional materials.


Oceanibaculum nanhaiense sp. nov., isolated from surface seawater.
A taxonomic study was carried out on strain L54-1-50T, which was isolated from surface seawater of the South China Sea. Cells of strain L54-1-50T were Gram-stain-negative, rod-shaped, oxidase-positive and catalase-positive. Growth was observed at salinities from 0 to 9 % (optimum 2 %, w/v), at pH 6.0-10.0 (optimum 8.0-9.0) and at temperatures from 10 to 45 °C (optimum 25-37 °C), but not at 4 or 50 °C. The 16S rRNA gene sequence analysis indicated that strain L54-1-50T was a member of the genus Oceanibaculum, related to Oceanibaculum indicum P24T (98.8 %) and Oceanibaculum pacificum MC2UP-L3T (97.7 %). The digital DNA-DNA hybridization values between strain L54-1-50T and the two type strains O. indicum P24T and O. pacificum MC2UP-L3T were 35.4±2.5 and 23.7±2.5 %, respectively. The average nucleotide identity values between strain L54-1-50T and two type strains were 79.7 and 88.3 %, respectively. The major cellular fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), C16 : 0 and C18 : 1 2-OH. The respiratory quinone was Q-10. The polar lipids comprised diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylglycerol, two unidentified phospholipids and three unidentified lipids. The G+C content of the chromosomal DNA was 65.1 mol%. The combined genotypic and phenotypic data showed that strain L54-1-50T represents a novel species of the genus Oceanibaculum, for which the name Oceanibaculumnanhaiense sp. nov. is proposed, with the type strain L54-1-50T (=KCTC 52312T=MCCC 1A05150T).


Are health states "timeless"? The case of the standard gamble method.
The standard gamble method, as currently recommended for use in health care program evaluation, provides an individual's preference score or "utility weight" for living in a given health state for the rest of the individual's life. Many researchers interpret this value as a time-independent or "timeless" one and order health states on a scale of zero (death) to one (full health), regardless of the time spent in the health state. This article examines whether preference scores for a severe pain health state are "timeless," or in other words whether the utility independence assumption is satisfied. Our study results suggest that for the majority of respondents, the preference scores are not independent of time.


Reprint of "Driving avoidance by older adults: Is it always self-regulation?".
Self-regulation shows promise as a means by which older adults can continue to drive at some level without having to stop altogether. Self-regulation is generally described as the process of modifying or adjusting one's driving patterns by driving less or intentionally avoiding driving situations considered to be challenging, typically in response to an awareness that driving skills have declined. However, most studies asking older adults whether they avoid certain driving situations or have reduced the amount of driving they do under certain circumstances have not delved deeper into the motivations for such avoidance or driving reduction. There are many reasons for modifying driving that have nothing to do with self-regulation, such as no longer needing to take trips at certain times of day because of changes in preferences or lifestyles. The purpose of this study was to examine self-regulatory practices among older adults at multiple levels of driver performance and decision making, taking into account the specific motivations for avoiding particular driving situations or engaging in other driving practices. Study participants completed a computer-based questionnaire on driving self-regulation. Results suggest that self-regulation is a complex process that cannot be defined simply by the reported driving modifications made by drivers. Understanding the motivations for these behaviors is necessary and the study showed that they are varied and differ considerably across driving situations. Reasons for driving avoidance or other practices were often more closely related to lifestyle or preferences than to self-regulation. Based on these findings, three distinct groups were identified with regard to whether and for what reasons participants modified their driving.


Digital Omicron detection using unscripted voice samples from social media.
The success of artificial intelligence in clinical environments relies upon the diversity and availability of training data. In some cases, social media data may be used to counterbalance the limited amount of accessible, well-curated clinical data, but this possibility remains largely unexplored. In this study, we mined YouTube to collect voice data from individuals with self-declared positive COVID-19 tests during time periods in which Omicron was the predominant variant1,2,3, while also sampling non-Omicron COVID-19 variants, other upper respiratory infections (URI), and healthy subjects. The resulting dataset was used to train a DenseNet model to detect the Omicron variant from voice changes. Our model achieved 0.85/0.80 specificity/sensitivity in separating Omicron samples from healthy samples and 0.76/0.70 specificity/sensitivity in separating Omicron samples from symptomatic non-COVID samples. In comparison with past studies, which used scripted voice samples, we showed that leveraging the intra-sample variance inherent to unscripted speech enhanced generalization. Our work introduced novel design paradigms for audio-based diagnostic tools and established the potential of social media data to train digital diagnostic models suitable for real-world deployment.


Central vasopressin system mediation of acute pressor effect of gamma-MSH.
Intravenous (iv) administration of gamma-melanocyte-stimulating hormone (gamma-MSH) produces central sympathetically mediated pressor and cardioaccelerator effects and increases the activity of hypothalamic vasopressinergic neurons. The autonomic actions are similar to infusion of vasopressin (Vp) into the hindbrain of 4th ventricle (Ven). To ascertain whether activation of the central Vp system is the proximate cause of the pressor effects of gamma-MSH, we investigated the effects of gamma-MSH in rats pre- and postblockade of central nervous system Vp receptors and in rats with a hereditary lack of vasopressin (Brattleboro strain). Central Vp receptor blockade significantly reduced (80%) the pressor effects of iv gamma-MSH. As a control, iv administration of the antagonist, while effective in blocking the pressor effect of iv Vp, had no effect on the gamma-MSH pressor response. When compared with their genetic controls (Long-Evans strain), Brattleboro rats also had greater than 80% reduction in their pressor response to iv gamma-MSH. The results indicate that circulating gamma-MSH activates the central Vp system to produce its sympathoexcitatory pressor effects.


Epidemiology of sudden cardiac death in Cameroon: the first population-based cohort survey in sub-Saharan Africa.
Incidence estimates of sudden cardiac death (SCD) in sub-Saharan Africa (SSA) are unknown. Over 12 months, the household administrative office and health community committee within neighbourhoods in two health areas of Douala, Cameroon, registered all deaths among 86 188 inhabitants aged >18 years. As part of an extended multi-source surveillance system, the Emergency Medical Service (EMS), local medical examiners and district hospital mortuaries were also surveyed. Whereas two physicians investigated every natural death, two cardiologists reviewed all unexpected natural deaths. There were 288 all-cause deaths and 27 (9.4%) were SCD. The crude incidence rate was 31.3 [95% confidence interval (CI): 20.3-40.6]/100 000 person-years. The age-standardized rate by the African standard population was 33.6 (95% CI: 22.4-44.9)/100 000 person-years. Death occurred at night in 37% of cases, including 11% of patients who died while asleep. Out-of-hospital sudden cardiac arrest occurred in 63% of cases, 55.5% of which occurred at home. Of the 88.9% cases of witnessed cardiac arrest, 63% occurred in the presence of a family member and cardiopulmonary resuscitation was attempted only in 3.7%. The burden of SCD in this African population is heavy with distinct characteristics, whereas awareness of SCD and prompt resuscitation efforts appear suboptimal. Larger epidemiological studies are required in SSA in order to implement preventive measures, especially in women and young people.


Further studies on the use of enzyme profiles to monitor residue accumulation in wildlife: plasma enzymes in starlings fed graded concentrations of Morsodren, DDE, Aroclor 1254 and malathion.
Wild-trapped starlings (Sturnus vulgaris) were fed concentrations of Morsodren (2, 4, and 8 ppm), DDE or Aroclor 1254 (5, 25, and 100 ppm), or malathion (8, 35, and 160 ppm) that were found to be sublethal in pen-reared Coturnix quail fed these amounts for 12 weeks. Plasma enzymes had to be measured earlier than planned in starlings fed Morsodren (at three weeks) or the organochlorine compounds (at seven weeks) because of unexpected, subsequent mortality. Variations in enzyme response were greater in wild than in pen-reared birds, but not enough to mask the toxicant-induced changes in enzyme activity. Cholinesterase activities decreased in birds fed Morsodren or malathion, and increased in those fed the organochlorine compounds. Lactate dehydrogenase activities increased two-fold in starlings fed Morsodren and two- to four-fold in those fed the organochlorine compounds, but only 50% in those fed malathion. Further examination of enzyme profiles showed that creatine kinase and aspartate aminotransferase activities increased two- to four-fold in birds fed Morsodren or the organochlorine compounds but not at all in those fed malathion. Thus the classes of environmental contaminants fed to starlings could be easily distinguished by these enzymatic parameters. Evaluation of enzymatic profiles appears to be a potentially valuable technique to monitor the presence of toxicants in wild populations, especially if used to complement standard chemical residue analyses. Here the residue analyses showed, after three weeks feeding, that mercury in the carcasses reflected the concentrations fed daily, whereas accumulation in the livers was two- to four-fold greater. After seven weeks feeding, liver residues of either organochlorine compound were about three-fold higher than the concentrations fed daily. However, four times as much DDE as Aroclor 1254 had accumulated in the carcasses.


Contrastive Learning for Preoperative Early Recurrence Prediction of Hepatocellular Carcinoma with Liver CT Image and Tumor Mask.
High early recurrence (ER) rate is the main factor leading to the poor outcome of patients with hepatocellular carcinoma (HCC). Accurate preoperative prediction of ER is thus highly desired for HCC treatment. Many radiomics solutions have been proposed for the preoperative prediction of HCC using CT images based on machine learning and deep learning methods. Nevertheless, most current radiomics approaches extract features only from segmented tumor regions that neglect the liver tissue information which is useful for HCC prognosis. In this work, we propose a deep prediction network based on CT images of full liver combined with tumor mask that provides tumor location information for better feature extraction to predict the ER of HCC. While, due to the complex imaging characteristics of HCC, the image-based ER prediction methods suffer from limited capability. Therefore, on the one hand, we propose to employ supervised contrastive loss to jointly train the deep prediction model with cross-entropy loss to alleviate the problem of intra-class variation and inter-class similarity of HCC. On the other hand, we incorporate the clinical data to further improve the prediction ability of the model. Experiments are extensively conducted to verify the effectiveness of our proposed deep prediction model and the contribution of liver tissue for prognosis assessment of HCC.


Effect of myostatin depletion on weight gain, hyperglycemia, and hepatic steatosis during five months of high-fat feeding in mice.
The marked hypermuscularity in mice with constitutive myostatin deficiency reduces fat accumulation and hyperglycemia induced by high-fat feeding, but it is unclear whether the smaller increase in muscle mass caused by postdevelopmental loss of myostatin activity has beneficial metabolic effects during high-fat feeding. We therefore examined how postdevelopmental myostatin knockout influenced effects of high-fat feeding. Male mice with ubiquitous expression of tamoxifen-inducible Cre recombinase were fed tamoxifen for 2 weeks at 4 months of age. This depleted myostatin in mice with floxed myostatin genes, but not in control mice with normal myostatin genes. Some mice were fed a high-fat diet (60% of energy) for 22 weeks, starting 2 weeks after cessation of tamoxifen feeding. Myostatin depletion increased skeletal muscle mass ∼30%. Hypermuscular mice had ∼50% less weight gain than control mice over the first 8 weeks of high-fat feeding. During the subsequent 3 months of high-fat feeding, additional weight gain was similar in control and myostatin-deficient mice. After 5 months of high-fat feeding, the mass of epididymal and retroperitoneal fat pads was similar in control and myostatin-deficient mice even though myostatin depletion reduced the weight gain attributable to the high-fat diet (mean weight with high-fat diet minus mean weight with low-fat diet: 19.9 g in control mice, 14.1 g in myostatin-deficient mice). Myostatin depletion did not alter fasting blood glucose levels after 3 or 5 months of high-fat feeding, but reduced glucose levels measured 90 min after intraperitoneal glucose injection. Myostatin depletion also attenuated hepatic steatosis and accumulation of fat in muscle tissue. We conclude that blocking myostatin signaling after maturity can attenuate some of the adverse effects of a high-fat diet.


Social avoidance in policing: Associations with cardiovascular disease and the role of social support.
The purpose of this paper is to examine the association of social avoidance among police, cardiovascular disease (CVD) (metabolic syndrome (MetSyn)), and social support. Participants were officers from the Buffalo Cardio-Metabolic Occupational Police Stress study (n = 289). Social avoidance (defined as the tendency to avoid social contact) and other subscales from the Cook-Medley Hostility Scale were analyzed. The mean number of MetSyn components across tertiles of the Cook-Medley scales was computed using analysis of variance and analysis of covariance. Social support was measured with the Social Provisions Scale, categorized as high or low based on the median. The mean number of MetSyn components increased significantly across tertiles of social avoidance (1.51 ± 0.18, 1.52 ± 0.12, and 1.81 ± 0.12); the only Cook-Medley subscale that remained significantly associated with MetSyn following adjustment for age and gender. Participants high in social avoidance reported significantly lower social support (79.9 ± 8.5 vs 85.8 ± 8.6; p = 0.001). The study is cross-sectional and therefore precludes causality. The authors were unable to determine the direction of associations between social avoidance and MetSyn. The measure of social support was unidimensional, including only perceived support; additional types of social support measures would be helpful. This study suggests that occupational-based police social isolation is associated with health outcomes and lower support. Several suggestions are made which will help to improve communication between the police and public. Examples are the use of social media, training in communication techniques, and changing the police role to one of public guardians. Social avoidance is the least studied the Cook-Medley subscale associated with CVD. It is important for the health of officers to maintain a social connection with others.


Characterization of a balanced reciprocal translocation, rcp(9;11)(q27;q11) in cattle.
Cytogenetic analysis of a phenotypically normal young bull from Marchigiana breed revealed the presence of an abnormal karyotype. The observation of longer and smaller chromosomes than BTA1 and BTA29, respectively in all metaphases suggested the presence of a reciprocal translocation. RBG-banding confirmed this hypothesis revealing the involvement of BTA9 and BTA11. FISH analyses using cattle-specific BAC clones (474A12 and 293G09 for BTA9; 035D03 for BTA11) identified rcp(9;11)(q27;q11) in the two regions affected. Moreover analyses performed on both parents established the 'de novo' origin of the anomaly. Comparison with human homologue sequences (HSA6q24.3-->q25.3 for BTA9q27 and HSA2q11.1-->q12.1 for BTA11q11) revealed that both breakpoint regions are gene rich as up to date at least 200 genes have been localized in these regions. Thus, further analyses are required to identify the sequences disrupted by the breakpoints and to verify their consequences on rcp carrier phenotype.


Prevalence of genes for enterotoxins, toxic shock syndrome toxin 1 and exfoliative toxin among clinical isolates of Staphylococcus pseudintermedius from canine origin.
A total of 74 Staphylococcus pseudintermedius strains were isolated from the 99 clinical cases of canine pyoderma or chronic otitis in our veterinary teaching hospital during May 2006-February 2008. In this study, we examined the genetic distribution of staphylococcal pyogenic toxins such as staphylococcal enterotoxins A (sea), B (seb), C (sec), D (sed), E (see), and toxic shock syndrome toxin 1 (tst) as well as the previously characterized S. intermedius exfoliative toxin (siet) among those isolates. The polymerase chain reaction analyses with the toxin gene-specific primers revealed that 18 (24.3%) of 74 S. pseudintermedius isolates carried the sec genes, but none of the sea, seb, sed, see and tst genes. Further DNA sequencing analysis of the amplified sec genes revealed that they all belonged to the canine type C staphylococcal enterotoxin (SEC(canine) ) whose superantigenic activity has been demonstrated. In addition to the sec(canine) genes, our polymerase chain reaction results showed that all the 74 isolates carried the siet gene. Since both SEC(canine) and SIET toxins are known to be biologically active, it would be interesting to investigate how those toxins are involved in the pathogenesis of the canine diseases by S. pseudintermedius such as pyoderma or chronic otitis.


The uptake and metabolism of chylomicron-remnant lipids by rat liver parenchymal and non-parenchymal cells in vitro.
The uptake and metabolism of chylomicron-remnant lipids by individual liver cell types was examined by incubating remnants with monolayer cultures of hepatocytes, Kupffer cells, and endothelial cells from rat liver. Remnants were prepared in vitro from radiolabelled mesenteric-lymph chylomicra, utilizing either purified lipoprotein lipase from bovine milk, or plasma isolated from heparinized rats. The resulting particles contained [3H]phosphatidylcholine and cholesterol, and [14C]oleate in the acylglycerol, phospholipid, fatty-acid and cholesterol-ester fractions. The capacities of the three cell types for uptake of both [3H]lipids and [14C]lipids were determined to be, on a per-cell basis, in the order: Kupffer greater than hepatocytes greater than endothelial. The relative proportions of [3H]phospholipid and total [3H]cholesterol taken up by hepatocytes and non-parenchymal cells remained constant with time. The uptake of [14C]oleoyl lipids by all three cell types was slightly greater than that of the total [3H]cholesterol and [3H]phospholipid components. There was evidence of cholesterol-ester hydrolysis and turnover of [14C]oleate in the phospholipid fraction in hepatocytes and Kupffer cells, but not endothelial cells, over the first 2 h. With both remnant preparations, these observations indicate that significant differences exist between the three major liver cell types with respect to the uptake and metabolism of remnant lipid components.


Regulatory landscape of dietary supplements and herbal medicines from a global perspective.
The number of Individuals that use dietary supplements and herbal medicine products are continuous to increase in many countries. The context of usage of a dietary supplement varies widely from country-to-country; in some countries supplement use is just limited to general health and well-being while others permit use for medicinal purposes. To date, there is little consensus from country to country on the scope, requirements, definition, or even the terminology in which dietary supplement and herbal medicines categories could be classified. Transparent science-based quality standards for the ingredients across these regulatory frameworks/definitions becomes even more important given the international supply chain. Meanwhile, there has been a rapid advancement in emerging technologies and data science applied to the field. This review was conceived at the Global Summit on Regulatory Sciences that took place in Beijing on September 2018 (GSRS2018) which is organized by Global Coalition for Regulatory Science Research (GCRSR) that consists of the global regulatory agencies from over ten countries including the European Union. This review summarizes a significant portion of discussions relating to a longitudinal comparison of the status for dietary supplements and herbal medicines among the different national jurisdictions and to the extent of how new tools and methodologies can improve the regulatory application.


Hypoglycemia in Beckwith-Wiedemann syndrome.
Beckwith-Wiedemann syndrome (BWS) is an overgrowth syndrome associated with macrosomia, macroglossia, abdominal wall defects, hypoglycemia in the neonatal period and embryonal cancers of infancy and early childhood. The frequency of hypoglycemia in this population is between 30% and 50%. The majority of infants with hypoglycemia will be asymptomatic and have resolution of the hypoglycemia within the first 3 days of life. Less than 5% will have hypoglycemia beyond the neonatal period requiring either continuous feeding or a partial pancreatectomy. The cause of hypoglycemia is unclear, but direct and indirect evidence supports a hyperinsulinemia as the major factor. Recent identification of the majority of genes associated with BWS in the 11p15 region and the genotype of persistent hyperinsulinemia hypoglycemia of childhood also in the 11p15 region may provide a molecular basis for hypoglycemia in BWS, particularly for the occasional patients with hypoglycemia requiring a partial pancreatectomy. Detailed genotype phenotype evaluations are needed and should provide an insight as to why patients with BWS have hypoglycemia.


Biogenesis of C-glycosyl flavones and profiling of flavonoid glycosides in lotus (Nelumbo nucifera).
Flavonoids in nine tissues of Nelumbo nucifera Gaertner were identified and quantified by high-performance liquid chromatography with diode array detector (HPLC-DAD) and HPLC-electrospray ionization-mass spectrometry (HPLC-ESI-MSn). Thirty-eight flavonoids were identified; eleven C-glycosides and five O-glycosides were discovered for the first time in N. nucifera. Most importantly, the C-glycosyl apigenin or luteolin detected in lotus plumules proved valuable for deep elucidation of flavonoid composition in lotus tissues and for further utilization as functional tea and medicine materials. Lotus leaves possessed the significantly highest amount of flavonoids (2.06E3±0.08 mg 100 g(-1) FW) and separating and purifying the bioactive compound, quercetin 3-O-glucuronide, from leaves showed great potential. In contrast, flavonoids in flower stalks, seed coats and kernels were extremely low. Simultaneously, the optimal picking time was confirmed by comparing the compound contents in five developmental phases. Finally, we proposed the putative flavonoid biosynthesis pathway in N. nucifera.


The prevalence of suspected ventilator-associated pneumonia in Scottish intensive care units.
Ventilator-associated pneumonia is the most common healthcare-associated infection in mechanically ventilated patients. Despite this, accurate diagnosis of ventilator-associated pneumonia is difficult owing to the variety of criteria that exist. In this prospective national audit, we aim to quantify the existence of patients with suspected ventilator-associated pneumonia that would not be detected by our standard healthcare-associated infection screening process. Furthermore, we aim to assess the impact of tracheostomy insertion, subglottic drainage endotracheal tubes and chlorhexidine gel on ventilator-associated pneumonia rate. Of the 227 patients recruited, suspected ventilator-associated pneumonia occurred in 32 of these patients. Using the HELICS definition, 13/32 (40.6%) patients were diagnosed with ventilator-associated pneumonia (H-posVAP). Suspected ventilator-associated pneumonia rate was increased in our tracheostomy population, decreased in the subglottic drainage endotracheal tube group and unchanged in the chlorhexidine group. The diagnosis of ventilator-associated pneumonia remains a contentious issue. The formalisation of the HELICS criteria by the European CDC should allow standardised data collection throughout Europe, which will enable more consistent data collection and meaningful data comparison in the future. Our data add weight to the argument against routine oral chlorhexidine. The use of subglottic drainage endotracheal tubes in preventing ventilator-associated pneumonia is interesting and requires further investigation.


Fibrous and protoplasmic astrocytes are biochemically and developmentally distinct.
We have studied semithin frozen sections of developing and adult rat central nervous system (CNS) by indirect immunofluorescence in order to determine the antigenic phenotype of protoplasmic and fibrous astrocytes. Using antibodies against glial fibrillary acidic protein (GFAP) to identify astrocytes, we show that the great majority of fibrous astrocytes in adult optic nerve are labeled by the monoclonal antibody A2B5, while the great majority of protoplasmic astrocytes in adult cerebral cortex are not. Astrocytes located at the periphery of the adult optic nerve that form the glial limiting membrane are more like protoplasmic astrocytes than fibrous astrocytes in that they strain relatively weakly with anti-GFAP antiserum and are A2B5-. In the developing rat optic nerve, protoplasmic-like astrocytes appear at least one week before the first fibrous astrocytes can be detected. Taken together with our previous observations on astrocytes in suspensions and cultures of developing rat optic nerve (Raff, M.C., E.R. Abney, J. Cohen, R. Lindsay, and M. Noble (1983) J. Neurosci. 3: 1289-1300; Raff, M.C., R.H. Miller, and M. Noble (1983) Nature 303: 390-396), these results suggest that protoplasmic and fibrous astrocytes are distinct classes of glial cells that differ in their antigenic phenotype and developmental history, as well as in their morphology and location within the CNS.


Changes in the biochemical and nutrient composition of seafood due to ocean acidification and warming.
Ocean acidification and warming may threaten future seafood production, safety and quality by negatively impacting the fitness of marine species. Identifying changes in nutritional quality, as well as species most at risk, is crucial if societies are to secure food production. Here, changes in the biochemical composition and nutritional properties of the commercially valuable oysters, Magallana gigas and Ostrea edulis, were evaluated following a 12-week exposure to six ocean acidification and warming scenarios that were designed to reflect the temperature (+3 °C above ambient) and atmospheric pCO2 conditions (increase of 350-600 ppm) predicted for the mid-to end-of-century. Results suggest that O. edulis, and especially M. gigas, are likely to become less nutritious (i.e. containing lower levels of protein, lipid, and carbohydrate), and have reduced caloric content under ocean acidification and warming. Important changes to essential mineral composition under ocean acidification and warming were evident in both species; enhanced accumulation of copper in M. gigas may be of concern regarding consumption safety. In light of these findings, the aquaculture industry may wish to consider a shift in focus toward species that are most robust to climate change and less prone to deterioration in quality, in order to secure future food provision and socio-economic benefits of aquaculture.