Cerebrospinal fluid leak after microsurgical surgery in vestibular schwannomas via retrosigmoidal craniotomy.
Cerebrospinal fluid (CSF) leak is still a common complication in surgery of vestibular schwannoma, increasing morbidity and prolonging hospital stay. Our single center study was performed to determine the incidences of CSF leaks after microsurgical removal of vestibular schwannoma via a retrosigmoidal approach with two different surgical closure techniques. Between January 2003 and December 2009 in 81 patients, microsurgical tumor resection using a suboccipital, retrosigmoidal approach was performed with an interdisciplinary ENT and neurosurgical management was performed. In 41 cases, the dural closure was done using a sandwich technique: subdural closure with TissuFleece® respectively Spongostan®, and after that dural suture and epidural Tachosil® were fixed on. In 40 cases, the dura was sealed epidurally with Tachosil after suture. In 65 cases, the posterior wall of the petrous bone was drilled. The closure was performed using muscle and FibrinGlue®. All patients had a minimal follow-up of 1 year. Seven patients (8.6%) developed a CSF fistula. Three patients (3.7%) underwent surgical procedure because of persisting CSF fistula while in four cases (4.9%) spontaneous closure under lumbar drain was observed. Comparing the different techniques of dural sealing, we found in 41 patients with sandwich technique three CSF leaks (7.3%) while there were four CSF leaks (10%) in 40 patients with a single epidurally sealed dural closure (P=0.69). No rhinorrhea or otorhinorrhea was observed. No intracranial infection or meningitis in case of CSF leak occurred. Suture and occlusion of the dura is an important step to prevent CSF leak and postoperative infection. By comparing sandwich technique and single-layer dural sealing, no significant difference could be shown.


The Heart-health Associated Research, Dissemination and Intervention in the Community (HARDIC) Trial for Nepalese Mothers regarding Diet and Physical Activity: A Process Evaluation.
Background Mothers with young children in the peri-urban Jhaukhel-Duwakot Health Demographic Surveillance site of Bhaktapur district have misconceptions and poor behavioural practice regarding diet and physical activity. We developed the Hearthealth Associated Research, Dissemination and Intervention in the Community trial - a health promotion intervention for mothers. Objective The objective of this study was to evaluate the intervention's feasibility, acceptability, potential for transferability and scaling up, and to determine its immediate outcome. Method Duwakot and Jhaukhel were randomly selected as the intervention and control communities, respectively. We trained 47 peer mothers from Duwakot, each of whom gave classes with 10 fellow mothers of their neighbourhood. The process evaluation was carried out on a continuous basis at different points of the intervention held from August to November 2016. Result In round one, the participation and completion rates were both > 90% for peer mothers; and 85% and 70%, respectively, for the fellow mothers. However, the participation rates fell in the round two of the intervention. On the whole, the mothers expressed satisfaction and acceptance of the course content and training modality. Immediate evaluation of the intervention showed improvement of knowledge, attitude and practice of diet and physical activity among both groups of mothers. Conclusion The successful implementation of the intervention targeting diet and physical activity clearly demonstrates the feasibility of health promotional activities in the Nepalese community for improvement of cardiovascular health.


The Adipokine Network in Rheumatic Joint Diseases.
Rheumatic diseases encompass a diverse group of chronic disorders that commonly affect musculoskeletal structures. Osteoarthritis (OA) and rheumatoid arthritis (RA) are the two most common, leading to considerable functional limitations and irreversible disability when patients are unsuccessfully treated. Although the specific causes of many rheumatic conditions remain unknown, it is generally accepted that immune mechanisms and/or uncontrolled inflammatory responses are involved in their etiology and symptomatology. In this regard, the bidirectional communication between neuroendocrine and immune system has been demonstrated to provide a homeostatic network that is involved in several pathological conditions. Adipokines represent a wide variety of bioactive, immune and inflammatory mediators mainly released by adipocytes that act as signal molecules in the neuroendocrine-immune interactions. Adipokines can also be synthesized by synoviocytes, osteoclasts, osteoblasts, chondrocytes and inflammatory cells in the joint microenvironment, showing potent modulatory properties on different effector cells in OA and RA pathogenesis. Effects of adiponectin, leptin, resistin and visfatin on local and systemic inflammation are broadly described. However, more recently, other adipokines, such as progranulin, chemerin, lipocalin-2, vaspin, omentin-1 and nesfatin, have been recognized to display immunomodulatory actions in rheumatic diseases. This review highlights the latest relevant findings on the role of the adipokine network in the pathophysiology of OA and RA.


Characterization of caulobacters isolated from wastewater treatment systems.
Caulobacters are generally assumed to be found only in environments of low organic content; however, we readily isolated strains from a variety of sewage treatment system designs and locations, and 33 distinct strains were characterized. Most were morphologically similar, having the crescent-shaped cell body, short stalk, and hexagonally packed, paracrystalline surface (S) layer characteristic of several Caulobacter crescentus laboratory strains. Upon closer examination, they were distinguishable on the basis of protein band profiles on polyacrylamide gel electrophoresis, gross colony characteristics, or holdfast composition or by DNA restriction fragment length polymorphism analysis with flagellin and S-layer gene probes. Most of the isolates contained one or more high-molecular-weight plasmids and were resistant to a number of antibiotics, characteristics generally not shared with caulobacters isolated from other sources. Six of the 33 strains were retained because they did not fit the typical isolate profile; these strains are overrepresented in our collection compared with their relative proportion in wastewater treatment systems. By colony hybridization and restriction fragment length polymorphism analysis, all of these and one typical isolate showed less homology than the others to the surface array gene of a laboratory strain (C. crescentus CB15), and three hybridized less strongly with the flagellin gene from the same strain. In sum, although the strains were distinguishable, caulobacters from the wastewater treatment systems we examined were relatively homogenous, were similar to characterized laboratory strains, and, with exceptions, could probably be reliably detected as a group by gene probes derived from C. crescentus strains.


[Characteristics of the peripheral neuromotor pathology in Chernobyl accident clean-up personnel late after the accident].
Clinical and electromyographic (EMG) examinations of the Chernobyl wreckers exposed to radiation revealed radiological damage to peripheral nervous system. 10-15 years after the accident they complain of weakness, headache, dizziness, muscle pain. Late damage to the peripheral nervous system manifest with EMG-detected axonal and demyelinating lesions, compression injuries of the peripheral nerves (tunnel syndromes), involvement of spinal cord motor neurons. In wreckers with thyroid lesions tunnel syndromes occur more often.


Renal response to repetitive exposure to heme proteins: chronic injury induced by an acute insult.
Renal diseases are conventionally classified into acute and chronic disorders. We questioned whether acute, reversible, renal insults may be induced to incite a chronic scarring process, employing as an acute insult the glycerol model of heme protein-induced renal injury. Rats were subjected to weekly injections of hypertonic glycerol for up to six months. Renal function was serially determined, and the effect of such insults on renal histology and renal expression of collagen and fibrogenic cytokines was assessed. After the first injection of glycerol, which, expectedly, induced a prompt fall in the glomerular filtration rate (GFR), subsequent injections encountered a remarkable renal resistance in that the fall in GFR was markedly blunted. This resistance to acute decline in renal function in rats subjected to repetitive injections of glycerol was accompanied by less necrosis and apoptosis of renal tubular epithelial cells after such injections. The attenuation in the fall in GFR in response to repetitive exposure to glycerol-induced heme protein injury was maintained for up to six months. A progressive decline in GFR appeared after three months and was accompanied by histologic tubulointerstitial injury, the latter assessed at six months. These kidneys demonstrated up-regulation of collagen I, III, and IV in conjunction with increased expression of the oxidant-inducible, chemotactic cytokine, monocyte chemoattractant protein-1 (MCP-1), and the oxidant-inducible, fibrogenic cytokine, transforming growth factor-beta1 (TGF-beta1). The exposure of the kidney to a single injection of hypertonic glycerol increased the expression of both cytokines some three to five days following this exposure, while the exposure of NRK 49F cells in culture to an iron-dependent model of oxidative stress also increased expression of TGF-beta1 and collagen mRNAs. We conclude that this nephrotoxic insult, repetitively administered, encounters a resistance in the kidney such that the expected fall in GFR does not occur. However, with time, such resistance is accompanied by a decrease in GFR, the latter associated with chronic tubulointerstitial disease. Thus, a long-term cost is exacted, either along with, or as a consequence of, such resistance. We suggest that chronic up-regulation of such oxidant-inducible genes such as TGF-beta1 and MCP-1 contributes to tubulointerstitial disease, and iron-mediated oxidative stress may directly induce TGF-beta1.


Buprenorphine mortality, seizures and prescription data in the UK, 1980-2002.
Buprenorphine safety in overdose has been debated recently, but no mortality data related to this compound from the UK have been published. To gather together all of the buprenorphine mortality figures, a number of different sources have been checked. To inform on buprenorphine safety issues, accessible information related to its availability indicators (i.e. prescriptions; seizures) data for the 1980-2002 time frame have been sought. In the UK, during this period, buprenorphine was mentioned in 43 fatalities. Typically, victims were males in the 25-44 age group. In 12 cases (28% of total), a verdict of suicide was given. Buprenorphine was detected on its own in seven cases; more frequently, it was found together with benzodiazepines and other opiates. Large quantities of buprenorphine were prescribed both in England in 1985-1989 and in 1991-1992 in Scotland, where seizures reached their highest levels. Buprenorphine prescriptions seemed to peak again after 1999, when high dose buprenorphine formulations entered the UK market. No positive correlation was found between the number of buprenorphine deaths over the years and either buprenorphine dispensings/prescriptions or seizures. However, an increase in buprenorphine-related deaths since 1999 was identified and this may be an issue which should be carefully monitored over the next few years.


[Intramolecular isotope distribution of metabolites in the glycolytic chain].
Based on the mechanism of the fractionation of carbon isotopes in the glycolytic chain proposed earlier, a method for the qualitative prediction of intramolecular isotope distribution of metabolites synthesized through the glycolytic chain and conjugated reactions was developed. Among them are fatty acids, monoterpenes, hopanoids, branched amino acids, amino acids of the family of trioses, metabolites synthesized by the shikimate pathway, metabolites of the Krebs cycle, amino acids of the family of ketoglutaric and oxalacetic acids. The method was tested by comparing the natural and theoretically predicted isotope distributions of carbon. The model for the description of theoretical isotope distributions is based on oscillations in the glycolytic chain. In the glycolysis phase, an isotope effect of decarboxylation of pyruvate arises, which determines the experimentally observed isotope inhomogeneity of metabolites. The isotope distributions of carbon depend on the sequence of syntheses of metabolites in the cell cycle, which corresponds to the degrees of exhaustion of the pyruvate depot that supplies structural units for the synthesis. In repeated cycles, the syntheses of metabolites occur in phase in addition to the carbon flux passing through the glycolytic chain, a carbon flux entering from the photorespiratory chain is involved in the synt, which is enriched in carbon heavy isotope. This flux is used, in particular, for the formation of carbon fragments of complex molecules.


Repeated thermal conditioning during the neonatal period affects behavioral and physiological responses to acute heat stress in chicks.
The aim of the present study was to investigate the effects of repeated thermal conditioning (RTC) at an early age on physiological and behavioral responses in chicks. Birds were assigned to one of the four treatments in which the RTC was exposure to 40 °C for 15 min daily. The treatments were 1) no thermal conditioning (control); 2) early exposure group (EE; RTC from 2 to 4 days of age); 3) later exposure group (LE; RTC from 5 to 7 days of age); or 4) both early and later exposure (BE; RTC from 2 to 7 days of age). All groups of chicks were challenged with high ambient temperature (40 °C for 15 min) at two weeks of age. During heat challenge, initiation times of dissipation behaviors (panting and wing-drooping) were measured. Rectal temperature and respiration rate were measured after and before heat challenge. Hypothalamic samples and blood were collected at the end of heat challenges. Initiation times of dissipation behaviors and rectal temperature were not affected by the treatments. Increases in respiration rate in response to heat challenge were suppressed by early RTC treatment. There was no clear pattern of glucose levels in relation to thermal conditioning, whereas plasma corticosterone levels were decreased by early treatment (EE and BE groups). Hypothalamic thyrotropin releasing hormone gene expression was suppressed by early and later thermal conditioning and suppressed further by both early and later exposure. Neuropeptide Y gene expression in the BE group was lower than in the other groups, with a similar trend for corticotropin releasing hormone expression. Our results suggest that the effect of repeated thermal conditioning on the central thermoregulatory system depends on the number of times that chicks experienced conditioning. In addition, repeated thermal conditioning has greater effects on the acquisition of thermotolerance when conditioning occurs in chicks of two to four days of age in comparison with chicks of five to seven days of age.


Construct validity of a 12-item WOMAC for assessment of femoro-acetabular impingement and osteoarthritis of the hip.
Evaluation of the internal construct validity of the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index adapted for use in patients with femoro-acetabular impingement (FAI) and osteoarthritis (OA) of the hip. Distribution of a German version of WOMAC to patients upon first consultation. Patients with FAI [n=100, mean age 31.7 years, standard deviation (SD) 9.7] and OA (n=57, mean age 60.3 years, SD 11.7) and without comorbidities or prior hip surgery were included and compared to age- and gender-matched control population to FAI (n=200, mean age 32.6 years, SD 5.6). WOMAC data of 157 questionnaires were evaluated by Rasch analysis using RUMM2020 software. Summation of total WOMAC shows misfit to the Rasch model as well as multidimensionality. While the pain subset shows adequate fit and is unidimensional, item reduction is required to fit a unidimensional subset of functional items to the Rasch model. Summating the two fitting subsets yields again slight model misfit and multidimensionality requiring further item reduction. Finally, a 12-item version of the total WOMAC shows good model fit and unidimensionality, i.e., internal construct validity, for assessment of patients with FAI and OA without differential item functioning (DIF). A person separation index (PSI)=0.93 indicates a high internal consistency reliability for the 12-item subscale. Scores for FAI are significantly higher than control (P<0.001, effect size 0.71) and lower than OA group (P<0.001, effect size 0.45). Adequate statistical power is shown discriminating the three groups, therefore indicating some evidence also for external construct validity. The WOMAC as a total construct is multidimensional and summating the subsets into a total score is not valid. The reduced 12-item WOMAC is demonstrated to have internal construct validity for assessing patients with FAI and OA on the same scale and high internal consistency reliability. Discrimination of the groups with adequate statistical power also indicates external construct validity.


Friends Turned Foes: Angiogenic Growth Factors beyond Angiogenesis.
Angiogenesis, the formation of new blood vessels from pre-existing ones is a biological process that ensures an adequate blood flow is maintained to provide the cells with a sufficient supply of nutrients and oxygen within the body. Numerous soluble growth factors and inhibitors, cytokines, proteases as well as extracellular matrix proteins and adhesion molecules stringently regulate the multi-factorial process of angiogenesis. The properties and interactions of key angiogenic molecules such as vascular endothelial growth factors (VEGFs), fibroblast growth factors (FGFs) and angiopoietins have been investigated in great detail with respect to their molecular impact on angiogenesis. Since the discovery of angiogenic growth factors, much research has been focused on their biological actions and their potential use as therapeutic targets for angiogenic or anti-angiogenic strategies in a context-dependent manner depending on the pathologies. It is generally accepted that these factors play an indispensable role in angiogenesis. However, it is becoming increasingly evident that this is not their only role and it is likely that the angiogenic factors have important functions in a wider range of biological and pathological processes. The additional roles played by these molecules in numerous pathologies and biological processes beyond angiogenesis are discussed in this review.


[Dopamine receptors agonists (mirapex) in the treatment of Parkinson's disease].
Presented are our own results, along with literature data on dopamine receptors agonists, of Parkinson's disease (PD) therapy with mirapex. Thirty patients with PD (mean age 61,8 +/- 7,7 years, illness duration 8,4 +/- 1,3 years) were treated with mirapex in a mean dose 3,5 +/- 1,1 mg/day and levadopa. Mirapex efficacy was assessed with a number of quantitative scales. Mirapex improved patient's global state, movement and daily activity, and quality of life. The authors drew attention to the decrease of motor fluctuations expression and dyskinesias as well as anxiety, depression, cognitive dysfunction. The combination of high efficacy and good tolerability of mirapex is stressed.


Leaf meristems: an easily ignored component of the response to human disturbance in alpine grasslands.
Grazing and fencing are two important factors that influence productivity and biomass allocation in alpine grasslands. The relationship between root (R) and shoot (S) biomass and the root:shoot ratio (R/S) are critical parameters for estimating the terrestrial carbon stocks and biomass allocation mechanism responses to human activities. Previous studies have often used the belowground:aboveground biomass ratio (M b/M a) to replace the R/S in alpine ecosystems. However, these studies may have neglected the leaf meristem biomass, which belongs to the shoot but occurs below the soil surface, leading to a significant overestimation of the R/S ratio. We conducted a comparative study to explore the differences between the R/S and M b/M a at both the species (Stipa purpurea, Carex moorcroftii, and Artemisia nanschanica) and community levels on a Tibetan alpine grassland with grazing and fencing management blocks. The results revealed that the use of the M b/M a to express the R/S appeared to overestimate the actual value of the R/S, both at species and community levels. For S. purpurea, the M b/M a was three times higher than the R/S. The M b/M a was approximately two times higher than the R/S for the species of C. moorcroftii and A. nanschanica and at the community level. The relationships between the R-S and M b-M a exhibited different slopes for the alpine plants across all the management practices. Compared to the fenced grasslands, the plants in the grazing blocks not only allocated more biomass to the roots but also to the leaf meristems. The present study highlights the contribution of leaf meristems to the accurate assessment of shoot and belowground biomasses. The R/S and M b/M a should be cautiously used in combination in the future research. The understanding of the distinction between the R-S and M b-M a may help to improve the biomass allocation mechanism response to human disturbances in an alpine area.


Mapping brain activity at scale with cluster computing.
Understanding brain function requires monitoring and interpreting the activity of large networks of neurons during behavior. Advances in recording technology are greatly increasing the size and complexity of neural data. Analyzing such data will pose a fundamental bottleneck for neuroscience. We present a library of analytical tools called Thunder built on the open-source Apache Spark platform for large-scale distributed computing. The library implements a variety of univariate and multivariate analyses with a modular, extendable structure well-suited to interactive exploration and analysis development. We demonstrate how these analyses find structure in large-scale neural data, including whole-brain light-sheet imaging data from fictively behaving larval zebrafish, and two-photon imaging data from behaving mouse. The analyses relate neuronal responses to sensory input and behavior, run in minutes or less and can be used on a private cluster or in the cloud. Our open-source framework thus holds promise for turning brain activity mapping efforts into biological insights.


Sarcopenia in a mice model of chronic liver disease: role of the ubiquitin-proteasome system and oxidative stress.
Sarcopenia is the loss of muscle mass and strength produced by aging or secondary to chronic diseases such as chronic liver disease (CLD). Although not all types of sarcopenia involve the same features, the most common are decreased fiber diameter and myosin heavy chain (MHC) levels, increased activity of ubiquitin-proteasome system (UPS) and reactive oxygen species (ROS). In this study, we aim to characterize the development of sarcopenia secondary to CLD induced by the hepatotoxin 5-diethoxycarbonyl-1,4-dihydrocollidine (DDC). For this purpose, four-months-old male C57BL6 mice were fed with normal diet or DDC supplemented diet for 6 weeks. Functional tests to evaluate muscle strength, mobility, and motor skills were performed in alive mice. The muscle strength in isolated gastrocnemius was also assayed via electrophysiological measurements. Morphometric measures of fibers' diameter, total and ubiquitinated protein levels of myosin heavy chain (MHC), E3 ubiquitin ligases, ROS, and oxidation-dependent modified proteins in gastrocnemius tissue were also determined. Our results demonstrated that mice fed the DDC diet developed muscle wasting as evidenced by a loss of muscle mass and decreased muscle strength. The muscles of mice fed with DDC diet have a decreased diameter of fibers and MHC levels, also as increased MuRF-1 and atrogin-1 protein levels, ROS levels, and oxidation-modified protein levels. Additionally, control and DDC mice have the same food and water intake as well as mobility. Our results demonstrate mice with CLD develop sarcopenia involving decreased levels of myofibrillar proteins, increased UPS, and oxidative stress, but not for impaired caloric intake or immobility.


Air Pollution Emissions 2008-2018 from Australian Coal Mining: Implications for Public and Occupational Health.
Occupational exposure limits for respirable coal dust are based on exposure during working hours, but coal miners may experience additional community-based exposures during nonworking hours. We analyzed Australia National Pollutant Inventory (NPI) data for the years 2008-2018 to estimate air pollutants (metals, nitrogen oxides, particulate matter ≤ 10 micrometers (PM10) and ≤2.5 micrometers (PM2.5)) originating from coal mines. PM10 levels from community-based air monitors in Queensland and New South Wales were also compared between mining and nonmining communities. Results indicated that tons of coal mined increased over the study period, and that levels of particulate matter, metals, and nitrogen oxides increased significantly over time as well. Coal mines accounted for 42.1% of national PM10 air emissions from NPI sites. PM2.5 from coal mines accounted for 19.5% of the national total, metals for 12.1%, and nitrogen oxides for 10.1%. Coal mining occurred in 57 different post codes; the 20 coal-mining post codes with the highest PM10 emissions were home to 160,037 people. Emissions of all studied pollutants were significantly higher from coal mining sites than from other types of NPI sites. Results from community-based air monitoring stations indicated significantly higher population PM10 exposure in coal mining communities than in nonmining communities. The health of the public at large is impacted by coal mining, but to the extent that miners also live near coal mining operations, their total exposure is underestimated by consideration of exposure only during working hours.


Lipopolysaccharides elicit an oxidative burst as a component of the innate immune system in the seagrass Thalassia testudinum.
This study represents the first report characterizing the biological effects of a lipopolysaccharide (LPS) immune modulator on a marine vascular plant. LPS was shown to serve as a strong elicitor of the early defense response in the subtropical seagrass Thalassia testudinum Banks ex König and was capable of inducing an oxidative burst identified at the single cell level. The formation of reactive oxygen species (ROS), detected by a redox-sensitive fluorescent probe and luminol-based chemiluminescence, included a diphenyleneiodonium sensitive response, suggesting the involvement of an NADPH oxidase. A 900 bp cDNA fragment coding for this enzyme was sequenced and found to encode a NAD binding pocket domain with extensive homology to the Arabidopsis thaliana rbohF (respiratory burst oxidase homolog) gene. The triggered release of ROS occurred at 20 min post-elicitation and was dose-dependent, requiring a minimal threshold of 50 μg/mL LPS. Pharmacological dissection of the early events preceding ROS emission indicated that the signal transduction chain of events involved extracellular alkalinization, G-proteins, phospholipase A2, as well as K(+), Ca(2+), and anion channels. Despite exclusively thriving in a marine environment, seagrasses contain ROS-generating machinery and signal transduction components that appear to be evolutionarily conserved with the well-characterized defense response systems found in terrestrial plants.


Duration of dual antiplatelet therapy and associated outcomes following percutaneous coronary intervention for acute myocardial infarction: contemporary practice insights from the Canadian Observational Antiplatelet Study.
There is a paucity of real-world, contemporary data of practice patterns and clinical outcomes following dual-antiplatelet therapy (DAPT) in acute myocardial infarction (AMI) patients treated with percutaneous coronary intervention (PCI). The Canadian Observational Antiplatelet Study was a prospective, multicentre, cohort study examining adenosine diphosphate receptor antagonist use following PCI for AMI. We compared practice patterns, patient characteristics, and clinical outcomes in relation to DAPT duration (<6 weeks, 6 weeks to <6 months, 6 to <12, and ≥12 months). The primary outcome was the composite of non-fatal AMI, unplanned coronary revascularization, stent thrombosis, new or worsening heart failure, cardiogenic shock, or stroke. We identified 2034 patients with AMI treated with PCI. DAPT duration was <6 weeks in 5.2% of patients; 6 weeks to <6 months in 7.0%; 6 to <12 months in 12.6%; and ≥12 months in 75.3%. Patients who discontinued DAPT early had higher GRACE risk scores. Overall, mortality rate at 15 months was 2.5%. Compared with a duration of DAPT of ≥12 months, discontinuation of DAPT <6 weeks (P < 0.0001) and 6 weeks to <6 months (P = 0.02), but not 6 months to <12 months (P = 0.06), were independently associated with a higher incidence of the primary outcome among survivors. One-in-four patients with AMI treated with PCI discontinued DAPT prior to the guideline-recommended 12-month duration. Patients in whom DAPT was discontinued early were at higher baseline risk and had higher rates of non-fatal ischaemic events during follow up.


Ten-Year Secular Trends in Sleep/Wake Patterns in Shanghai and Hong Kong School-Aged Children: A Tale of Two Cities.
To compare the secular trends of sleep/wake patterns in school-aged children in Hong Kong and Shanghai, two major metropolitan cities in China with two different policies that school start time was delayed in Shanghai, but advanced in Hong Kong in 10 years' time. Participants were from two waves of cross-sectional school-based surveys of children aged 6 to 11 years. In Shanghai, 4,339 and 13,795 children participated in the 2005 and 2014 surveys, respectively. In Hong Kong, 6,231 and 4,585 children participated in the 2003 and 2012 surveys, respectively. Parents reported their children's bedtime and wakeup time, and thus sleep duration, short sleep (≤ 9 hours) and weekend oversleep (difference in sleep duration between weekday and weekend > 2 hours) were determined. Hong Kong children had later bedtime and wakeup time and slept consistently less than their Shanghai counterparts at both survey time points. The shorter sleep duration was particularly marked during weekdays. Over the interval period, weekday sleep duration significantly decreased from 9.2 to 8.9 hours as wakeup time became earlier for Hong Kong children, but increased from 9.4 to 9.6 hours as wakeup time became later for children in Shanghai. Children from both cities slept longer on the weekends. Prevalence of weekend oversleep significantly increased in Hong Kong children, but no interval change was found in Shanghai children. The findings indicate subcultural differences in sleep/wake patterns in Shanghai and Hong Kong school-aged children. In particular, sleep duration had increased for Shanghai children, but decreased for Hong Kong children over 10 years. The benefits and barriers of delaying school start time for optimizing sleep health in school-aged children should be further explored.


Variable temperature thin film indentation with a flat punch.
We present modifications to conventional nanoindentation that realize variable temperature, flat punch indentation of ultrathin films. The technique provides generation of large strain, thin film extrusion of precise geometries that idealize the essential flows of nanoimprint lithography, and approximate constant area squeeze flow rheometry performed on thin, macroscopic soft matter samples. Punch radii as small as 185 nm have been realized in ten-to-one confinement ratio testing of 36 nm thick polymer films controllably squeezed in the melt state to a gap width of a few nanometers. Self-consistent, compressive stress versus strain measurements of a wide variety of mechanical testing conditions are provided by using a single die-sample system with temperatures ranging from 20 to 125 degrees C and loading rates spanning two decades. Low roughness, well aligned flat punch dies with large contact areas provide precise detection of soft surfaces with standard nanoindenter stiffness sensitivity. Independent heating and thermometry with heaters and thermocouples attached to the die and sample allow introduction of a novel directional heat flux measurement method to ensure isothermal contact conditions. This is a crucial requirement for interpreting the mechanical response in temperature sensitive soft matter systems. Instrumented imprint is a new nanomechanics material testing platform that enables measurements of polymer and soft matter properties during large strains in confined, thin film geometries and extends materials testing capabilities of nanoindentation into low modulus, low strength glassy, and viscoelastic materials.


Escape from immune surveillance does not result in tolerance to tumor-associated antigens.
Despite expression of tumor-associated or tumor-specific antigens by most tumors, evasion of protective T-cell immunity is the rule rather than the exception. Understanding whether tumor immune escape primarily represents T-cell neglect, anergy/tolerance, or quantitative limits of an existent immune response is central to developing new strategies to enhance antitumor immunity. The authors studied the immune response to MB49, a tumor that naturally expresses HY. Immune surveillance was effective following low-dose tumor inocula, since normal female mice showed a diminished incidence and slower growth rate of MB49 compared with T-cell-depleted female mice and male mice. Following high-dose tumor inoculation, females developed large, progressive tumors but continued to demonstrate immune responses to class I and class II restricted HY epitopes. The HY reactive T cells remained capable of executing HY immune responses since T cells adoptively transferred from MB49-bearing animals mediated accelerated HY skin graft rejection compared with those taken from naive mice. Thus, MB49 does not induce immune tolerance to HY but rather escapes immune surveillance largely due to quantitative limits of the immune response. Treatment of tumor-bearing animals with rhIL7 significantly increased the number of T cells responding to HY but did not alter tumor growth rate. These results demonstrate that escape from immune surveillance does not necessarily imply immune tolerance to tumor antigens and that immunotherapy need not overcome tumor-induced tolerance per se, and suggest that substantial opportunities remain in tumor-bearing hosts to amplify weak but persistent antitumor immune responses.


Neudesin: a neuropeptide hormone decreased in subjects with polycystic ovary syndrome.
Neudesin is a neuropeptide hormone involves in female reproduction system via promoting effects of progesterone. Polycystic ovary syndrome (PCOS) is a metabolic and reproductive disorder associated with hypothalamic-pituitary-ovarian axis abnormalities and impaired negative feedback mechanism of progesterone upon gonadotropin-releasing hormone secretion. Our aims were to discover whether neudesin levels were altered in PCOS women comparing to controls and to determine the link of neudesin with hormonal-metabolic parameters in PCOS women. The current research was designed as a case-control study. Sixty-eight subjects with PCOS and 67 age- and body mass index (BMI)-matched subjects as controls were enrolled into the study. Circulating neudesin levels were measured by ELISA. Neudesin levels were significantly lower in PCOS subjects compared to controls (4.07 ± 1.22 vs. 6.02 ± 2.07 ng/ml, p < .001). Neudesin exhibited an inversely independent link with luteinizing hormone, free-androgen index, and BMI whereas it showed a positively independent link with progesterone in women with PCOS. Logistic regression analysis revealed that decreased neudesin levels were parallel with increased risk of having PCOS. Decreased neudesin levels were associated with hormonal disturbances in PCOS women, suggesting that neudesin may play a role in pathophysiology of PCOS.


Outpatient treatment costs and their potential impact on cancer care.
Cancer creates a tremendous financial burden. Cancer-related costs are categorized into direct, indirect, and psychosocial costs. Although there have been many reports on medical care costs, which are direct, those on other costs are extremely scarce. We estimated travel time and costs required for cancer patients to receive outpatient treatment. We studied 521 cancer patients receiving anti-cancer treatment between February 2009 and December 2012 at the Outpatient Chemotherapy Center of Teikyo University Chiba Medical Center. Address data were extracted from Data Warehouse electronic medical records, and travel distance and time required for outpatient treatment were calculated via MapInfo and ACT Distance Calculator Package. Transportation costs were estimated on the basis of ¥274 (=$3.00) per kilometer. The study design was approved by an ethics review board of Teikyo University (12-851). Average round-trip travel distance, time, and cost for all patients were 26.7 km, 72.5 min, and ¥7,303 ($79.99), respectively. Cancer patients incurred a travel cost of ¥4000-¥9000 ($40.00 to $100.00) for each outpatient treatment. With population aging, seniors living alone and senior households are increasing, and outpatient visits are becoming a common burden.


Fatigue Assessment from Facial Videos using Deep Neural Networks and Engineered Features Informed by Domain Knowledge.
Fatigue impairs cognitive and motor function, potentially leading to mishaps in high-pressure occupations such as aviation and emergency medical services. The current approach is primarily based on self-assessment, which is subjective and error-prone. An objective method is needed to detect severe and likely dangerous levels of fatigue quickly and accurately. Here, we present a quantitative evaluation tool that uses less than two minutes of facial video, captured using an iPad, to assess fatigue vs. alertness. The tool is fast, easy to use, and scalable since it uses cameras readily available on consumer-electronic devices. We compared the classification performance between a Long Short-Term Memory (LSTM) deep neural network and a Random Forest (RF) classifier applied to engineered features informed by domain knowledge. The preliminary results on an 11-subject dataset show that RF outperforms LSTM, with added interpretability on the features used. For the RF classifiers, the average areas under the receiver operating characteristic curve, based on the 11-fold and individualized 11-fold cross validations, are 0.72 ± 0.16 and 0.8 ± 0.12, respectively. Equal error rates are 0.34 and 0.26, respectively. This study presents a promising approach for rapid fatigue detection. Additional data will be collected to assess the generalizability across populations.


Intradermal hepatitis B vaccination in an abbreviated schedule.
Two low-dose intradermal regimens for hepatitis B vaccination were compared with the standard 1 ml dose administered intramuscularly to healthy, 22-42 year old individuals. All regimens were administered in an abbreviated time schedule. Nineteen individuals (ID-1 group) received three 0.1 ml (2 micrograms) doses intradermally at times 0, 1 month and 4 months. Twenty-four individuals (ID-2 group) received two injections of 0.2 ml (4 micrograms) each intradermally at time 0 and one 0.1 ml (2 micrograms) injection 4 months later. Twenty individuals (IM group) received the recommended three 1.0 ml (20 micrograms) doses intramuscularly at times 0, 1 month, and 4 months. No significant adverse reactions were attributable to the intradermal administration of vaccine although the majority of vaccinees developed small areas of induration and hyperpigmentation at the injection site that persisted for several months. One month following the last injection, all vaccinees had developed anti-HBsAg antibodies. One hundred percent of ID-1 and IM vaccinees and 95% of ID-2 vaccinees had protective levels of antibody (greater than or equal to 10 mIU ml-1). The geometric mean titre (GMT) for the IM group (2692 mIU ml-1) was somewhat higher than for the ID-1 (1230 mIU ml-1) and the ID-2 (851 mlU ml-1) groups, but the differences were not statistically significant. Since anti-HBs antibodies are thought to confer protection against hepatitis B, these results suggest that a shortened regimen of intradermal vaccine may be effective in healthy adults. However, no efficacy study has yet been done with intradermal hepatitis B vaccine.


Scanning the human genome at kilobase resolution.
Normal genome variation and pathogenic genome alteration frequently affect small regions in the genome. Identifying those genomic changes remains a technical challenge. We report here the development of the DGS (Ditag Genome Scanning) technique for high-resolution analysis of genome structure. The basic features of DGS include (1) use of high-frequent restriction enzymes to fractionate the genome into small fragments; (2) collection of two tags from two ends of a given DNA fragment to form a ditag to represent the fragment; (3) application of the 454 sequencing system to reach a comprehensive ditag sequence collection; (4) determination of the genome origin of ditags by mapping to reference ditags from known genome sequences; (5) use of ditag sequences directly as the sense and antisense PCR primers to amplify the original DNA fragment. To study the relationship between ditags and genome structure, we performed a computational study by using the human genome reference sequences as a model, and analyzed the ditags experimentally collected from the well-characterized normal human DNA GM15510 and the leukemic human DNA of Kasumi-1 cells. Our studies show that DGS provides a kilobase resolution for studying genome structure with high specificity and high genome coverage. DGS can be applied to validate genome assembly, to compare genome similarity and variation in normal populations, and to identify genomic abnormality including insertion, inversion, deletion, translocation, and amplification in pathological genomes such as cancer genomes.


Immune tolerance induced by donor antigen and cyclophosphamide in rat fetal small bowel transplantation.
Donor specific immune tolerance is thought to be the ideal state for the recipient after organ transplantation. The administration of donor antigens and cyclophosphamide has been reported to induce donor specific immune tolerance in heart or liver transplantation. However, the effectiveness of this method for small bowel transplantation has not yet been studied. We assessed the cyclophosphamide induced immune tolerance on rat fetal small bowel transplantation. Lewis rats (RT1(1), n=99) were used as recipients while either F344 (RT1(1), n=44) or WKAM (RT1(u), n=47) rats were used as donors. The combination of F344 and Lewis rats produces an immunologically low responder, while that of WKAM and Lewis rat produces a high responder. Bone marrow and spleen cells were harvested from the donor rats and 3x10(8)/kg of each were administrated to the recipient rats intravenously on day 0. Next, cyclophosphamide was given either divisionally or bolously. The fetal small bowel of the same strain as the donor was transplanted into the rectus muscle of the recipient abdominal wall on day 10. On day 17, all grafts were taken out and graft survival was thereafter evaluated. The body weight of recipient was also assessed. Most of the grafts (87.5%) survived in the F344-Lewis rat (low responder) combination using the divisional administration of 120 mg/kg of cyclophosphamide. Histologically, most of them showed the whole layers of the intestinal architectureto be well preserved. The weight loss of the recipient was minimal after divisional administration. In contrast, no graft survived in the WKAM-Lewis rat (high responder) combination. Immune tolerance is considered to be induced by the administration of donor specific antigen and cyclophosphamide in an immunologically low responder combination. Therefore, this method is expected to be useful as an adjuvant therapy and may also be able to reduce the dose of immunosuppressive agents in living-related clinical small intestinal transplantation.


[Our programmed headache medical record. Contribution to the systematic registration and processing of a larger amount of headache patients using the computer].
We present a detailed anterospective inventory on headache which has been developed systematically with several alterations in the course of the experience of several years. It offers the possibility of an almost complete collection of data which can be fed directly into a computer and it appears to be a valuable aid in the training of new co-workers. Some clinical results are discussed on the basis of the inventory.


Minimal clinically important differences: review of methods.
Determining a minimal clinically important difference to be detected in a clinical trial is a critical methodological step in the design of a study. In this review, the different methods that have been used in detecting important changes or differences are considered and categorized according to a classification system consisting of which groups are contrasted, the setting of the results, and the type of change of difference being quantified. It was found that most methods and procedures for deriving minimal clinically important differences considered important changes from the viewpoint of a group of patients. Development of methods that focus on individuals should be a goal of the future.


Associations of Trimester-Specific Exposure to Perchlorate, Thiocyanate, and Nitrate with Childhood Neurodevelopment: A Birth Cohort Study in China.
Studies about the impacts of maternal exposure to perchlorate, thiocyanate, and nitrate on offspring neurodevelopment are scarce. Based on a birth cohort in China, 1,028 mothers provided urine samples at three trimesters for determination of the three target analytes, and their offspring neurodevelopment was evaluated at 2 years old. Associations of maternal exposure to the three chemicals with offspring neurodevelopment were estimated using three statistical methods. Trimester-specific analyses using generalized estimating equation models showed that double increment of thiocyanate and nitrate during the first trimester was associated with 1.56 (95% CI: -2.82, -0.30) and 1.22 (-2.40, -0.03) point decreases in the offspring mental development index (MDI), respectively. Weighted quantile sum (WQS) regression analyses showed that the mixture exposure at the first and second trimesters was negatively associated with the offspring MDI (β = -2.39, 95% CI: -3.85, -0.93; β = -1.75, 95% CI: -3.04, -0.47, respectively) and thiocyanate contributed the most to the association (65.0 and 91.6%, respectively). Bayesian kernel machine regression analyses suggested an inverted U-shape relationship of maternal urinary thiocyanate with the offspring MDI. These findings suggested that prenatal exposure to the three chemicals (at current levels), especially thiocyanate and nitrate, may impair neurodevelopment. Early pregnancy seems to be the sensitive window.


Incorporating resident research into the dermatology residency program.
Programmatic changes for the dermatology residency program at The University of Texas Medical Branch were first introduced in 2005, with the faculty goal incorporating formal dermatology research projects into the 3-year postgraduate training period. This curriculum initially developed as a recommendation for voluntary scholarly project activity by residents, but it evolved into a program requirement for all residents in 2009. Departmental support for this activity includes assignment of a faculty mentor with similar interest about the research topic, financial support from the department for needed supplies, materials, and statistical consultation with the Office of Biostatistics for study design and data analysis, a 2-week elective that provides protected time from clinical activities for the purpose of preparing research for publication and submission to a peer-reviewed medical journal, and a departmental award in recognition for the best resident scholarly project each year. Since the inception of this program, five classes have graduated a total of 16 residents. Ten residents submitted their research studies for peer review and published their scholarly projects in seven dermatology journals through the current academic year. These articles included three prospective investigations, three surveys, one article related to dermatology education, one retrospective chart review, one case series, and one article about dermatopathology. An additional article from a 2012 graduate about dermatology education has also been submitted to a journal. This new program for residents was adapted from our historically successful Dermatology Honors Research Program for medical students at The University of Texas Medical Branch. Our experience with this academic initiative to promote dermatology research by residents is outlined. It is recommended that additional residency programs should consider adopting similar research programs to enrich resident education.


Process evaluation of a community-based adolescent obesity prevention project in Tonga.
The rising burden of obesity in Tonga is alarming. The promotion of healthy behaviours and environments requires immediate urgent action and a multi-sectoral approach. A three-year community based study titled the Ma'alahi Youth Project (MYP) conducted in Tonga from 2005-2008 aimed to increase the capacity of the whole community (schools, churches, parents and adolescents) to promote healthy eating and regular physical activity and to reduce the prevalence of overweight and obesity amongst youth and their families. This paper reflects on the process evaluation for MYP, against a set of Best Practice Principles for community-based obesity prevention. MYP was managed by the Fiji School of Medicine. A team of five staff in Tonga were committed to planning, implementation and evaluation of a strategic plan, the key planks of which were developed during a two day community workshop. Intervention activities were delivered in villages, churches and schools, on the main island of Tongatapu. Process evaluation data covering the resource utilisation associated with all intervention activities were collected, and analysed by dose, frequency and reach for specific strategies. The action plan included three standard objectives around capacity building, social marketing and evaluation; four nutrition; two physical activity objectives; and one around championing key people as role models. While the interventions included a wide mix of activities straddling across all of these objectives and in both school and village settings, there was a major focus on the social marketing and physical activity objectives. The intervention reach, frequency and dose varied widely across all activities, and showed no consistent patterns. The adolescent obesity interventions implemented as part of the MYP program comprised a wide range of activities conducted in multiple settings, touched a broad spectrum of the population (wider than the target group), but the dose and frequency of activities were generally insufficient and not sustained. Also the project confirmed that, while the MYP resulted in increased community awareness of healthy behaviours, Tonga is still in its infancy in terms of conducting public health research and lacks research infrastructure and capacity.


Tumours of the upper alimentary tract.
Tumours of the oropharynx of domestic animals are common in most parts of the world, but squamous cell carcinoma of the upper alimentary tract shows differences in prevalence in different geographical areas and occurs at different sites in the various species. Oral tumours of the melanogenic system are more common in dogs than in man. The following main histological categories, which broadly correspond to those used in the classification of tumours of man, are described: papilloma; squamous cell carcinoma; salivary gland tumours; malignant melanoma; tumours of soft (mesenchymal) tissues; tumours of the facial bones; tumours of haematopoietic and related tissues; and odontogenic tumours and jaw cysts. Papilloma, squamous cell carcinoma, malignant melanoma, fibroma, and fibrosarcoma account for about 80% of the tumours that occur in the upper alimentary tract of domestic animals.


Spontaneous binding of potential COVID-19 drugs (Camostat and Nafamostat) to human serine protease TMPRSS2.
Effective treatment or vaccine is not yet available for combating SARS coronavirus 2 (SARS-CoV-2) that caused the COVID-19 pandemic. Recent studies showed that two drugs, Camostat and Nafamostat, might be repurposed to treat COVID-19 by inhibiting human TMPRSS2 required for proteolytic activation of viral spike (S) glycoprotein. However, their molecular mechanisms of pharmacological action remain unclear. Here, we perform molecular dynamics simulations to investigate their native binding sites on TMPRSS2. We revealed that both drugs could spontaneously and stably bind to the TMPRSS2 catalytic center, and thereby inhibit its proteolytic processing of the S protein. Also, we found that Nafamostat is more specific than Camostat for binding to the catalytic center, consistent with reported observation that Nafamostat blocks the SARS-CoV-2 infection at a lower concentration. Thus, this study provides mechanistic insights into the Camostat and Nafamostat inhibition of the SARS-CoV-2 infection, and offers useful information for COVID-19 drug development.


Evaluation of color preference in zebrafish for learning and memory.
There is growing interest in using zebrafish (Danio rerio) as a model of neurodegenerative disorders such as Alzheimer's disease. A zebrafish model of tauopathies has recently been developed and characterized in terms of presence of the pathological hallmarks (i.e., neurofibrillary tangles and cell death). However, it is also necessary to validate these models for function by assessing learning and memory. The majority of tools to assess memory and learning in animal models involve visual stimuli, including color preference. The color preference of zebrafish has received little attention. To validate zebrafish as a model for color-associated-learning and memory, it is necessary to evaluate its natural preferences or any pre-existing biases towards specific colors. In the present study, we have used four different colors (red, yellow, green, and blue) to test natural color preferences of the zebrafish using two procedures: Place preference and T-maze. Results from both experiments indicate a strong aversion toward blue color relative to all other colors (red, yellow, and green) when tested in combinations. No preferences or biases were found among reds, yellows, and greens in the place preference procedure. However, red and green were equally preferred and both were preferred over yellow by zebrafish in the T-maze procedure. The results from the present study show a strong aversion towards blue color compared to red, green, and yellow, with yellow being less preferred relative to red and green. The findings from this study may underpin any further designing of color-based learning and memory paradigms or experiments involving aversion, anxiety, or fear in the zebrafish.


Electrodynamic assisted self-assembled fibrous hydrogel microcapsules: a novel 3D in vitro platform for assessment of nanoparticle toxicity.
Nanoparticle (NP) toxicity assessment is a critical step in assessing the health impacts of NP exposure to both consumers and occupational workers. In vitro assessment models comprising cells cultured in a two-dimensional tissue culture plate (2D-TCP) are an efficient and cost-effective choice for estimating the safety risks of NPs. However, in vitro culture of cells in 2D-TCPs distorts cell-integrin and cell-cell interactions and is not able to replicate an in vivo phenotype. Three-dimensional (3D) in vitro platforms provide a unique alternative to bridge the gap between traditional 2D in vitro and in vivo models. In this study, novel microcapsules of alginate hydrogel incorporated with natural polymeric nanofibers (chitin nanofibrils) and synthetic polymeric nanofibers poly(lactide-co-glycolide) are designed as a 3D in vitro platform. This study demonstrates for the first time that electrodynamic assisted self-assembled fibrous 3D hydrogel (3D-SAF hydrogel) microcapsules with a size in the range of 300-500 μm in diameter with a Young's modulus of 12.7-42 kPa can be obtained by varying the amount of nanofibers in the hydrogel precursor solutions. The 3D-SAF microcapsules were found to mimic the in vivo cellular microenvironment for cells to grow, as evaluated using A549 cells. Higher cellular spreading and prolonged proliferation of A549 cells were observed in 3D-SAF microcapsules compared to control microcapsules without the nanofibers. The 3D-SAF microcapsule integrated well plate was used to assess the toxicity of model NPs, e.g., Al2O3 and ZnO. The toxicity levels of the model NPs were found to be dependent on the chemistry of the NPs and their physical agglomeration in the test media. Our results demonstrate that 3D-SAF microcapsules with an in vivo mimicking microenvironment can be developed as a physiologically relevant platform for high-throughput toxicity screening of NPs or pharmaceutical drugs.


A history of Dwight D. Eisenhower's oral health.
This paper reviews the oral health history of President Eisenhower from his early years as a Lieutenant Colonel in the Tank Corps to the last years of his life. It includes first hand accounts of his oral care when President, provides evidence that his periodontitis stabilized after he quit smoking, documents the destructive effects of his parafunctional grinding on his dentition and how he was plagued with fracturing teeth the last fifteen years of his life. Finally, it shows the special care Eisenhower was given both as a renowned leader and a medically compromised patient.


The Willingness to Pay for a Quality Adjusted Life Year: A Review of the Empirical Literature.
There has been a rapid increase in the use of cost-effectiveness analysis, with quality adjusted life years (QALYs) as an outcome measure, in evaluating both medical technologies and public health interventions. Alongside, there is a growing literature on the monetary value of a QALY based on estimates of the willingness to pay (WTP). This paper conducts a review of the literature on the WTP for a QALY. In total, 24 studies containing 383 unique estimates of the WTP for a QALY are identified. Trimmed mean and median estimates amount to 74,159 and 24,226 Euros (2010 price level), respectively. In regression analyses, the results indicate that the WTP for a QALY is significantly higher if the QALY gain comes from life extension rather than quality of life improvements. The results also show that the WTP for a QALY is dependent on the size of the QALY gain valued. Copyright © 2014 John Wiley & Sons, Ltd.


The Plasmid-Borne tet(A) Gene Is an Important Factor Causing Tigecycline Resistance in ST11 Carbapenem-Resistant Klebsiella pneumoniae Under Selective Pressure.
The emergence and prevalence of tigecycline-resistant Klebsiella pneumoniae have seriously compromised the effectiveness of antimicrobial agents in the treatment of infections. To explore the role of the plasmid-borne tet(A) gene in tigecycline resistance in carbapenem-resistant K. pneumoniae (CRKP), a total of 63 CRKP isolates were collected from a tertiary hospital in Hangzhou, China. The minimum inhibitory concentration (MIC) of tigecycline, mutation rate of tet(A) gene, genetic surroundings of tet(A)-carrying transmissible plasmid and the contribution of tet(A) mutation to tigecycline resistance were analyzed using antimicrobial susceptibility test, whole-genome sequencing, tigecycline resistance evolution experiment, and plasmid conjugation experiment. Our results showed that 52.4% (33 isolates) of the test isolates carried the tet(A) gene; among them, 75.8% (25 isolates) exhibited a tigecycline non-susceptible phenotype (MIC = 4 mg/L). Three clonal groups (cluster I, cluster II, and cluster III) were identified in these tet(A)-bearing isolates. All 17 isolates belonged to serotype KL21 (cluster I), which differed by only 13 SNPs, suggesting a clonal spread of tet(A)-positive ST11 K. pneumoniae with serotype KL21 occurred in the sampling hospital. The induction of tigecycline resistance experiments showed that 71.4% of strains evolved tet(A) mutations and developed a high-level tigecycline resistance. Eight amino acid substitutions were identified in these mutants. The most common amino acid substitution was A370V, followed by S251A and G300E. Twelve isolates carrying tet(A) mutants succeeded in the filter mating experiment with a conjugation efficiency of 10-3-10-8. Tigecycline MICs in E. coli EC600 transconjugants with a mutated tet(A) were 2 to 8-fold higher than those in E. coli EC600 transconjugants with a wild-type tet(A). One ColRNAI/IncFII type and two IncFII type tet(A)-bearing conjugative plasmids were identified in this study, including a class 1 integron containing multiple antibiotic resistance genes, i.e., tet(A), qnrS1, bla LAP- 2, catA2, sul2, and dfrA14. Our study revealed the wide-spread situation of plasmid-borne tet(A) gene in clinical CRKP, and mutation of tet(A) is a potential driven force that lead to tigecycline resistance.


Molecular Pathways: Targeting MYC-induced metabolic reprogramming and oncogenic stress in cancer.
MYC is a multifunctional transcription factor that is deregulated in many human cancers. MYC impacts a collaborative genetic program that orchestrates cell proliferation, metabolism, and stress responses. Although the progression of MYC-amplified tumors shows robust dependence on MYC activity, directly targeting MYC as a therapeutic method has proven to be technically difficult. Therefore, alternative approaches are currently under development with a focus on interference with MYC-mediated downstream effects. To fuel rapid cell growth, MYC reprograms cancer cell metabolism in a way that is substantially different from normal cells. The MYC-induced metabolic signature is characterized by enhanced glucose and glutamine uptake, increased lactate production, and altered amino acid metabolism. Targeting MYC-reprogrammed cancer cell metabolism is considered to be promising based on multiple preclinical studies. In addition, the increased biosynthetic demand of MYC-driven tumors coupled with limited nutrient access within tumor microenvironments create multiple levels of oncogenic stress, which can also be used as tumor-specific targets for pharmacologic intervention. Presumably, the best therapeutic strategy for treating MYC-amplified tumors is combined targeting of multiple MYC-mediated pathways, especially those involved in regulating cell proliferation, metabolism, and oncogenic stress.


Change in the carotenoid and antioxidant content of spice red pepper (paprika) as a function of ripening and some technological factors.
A study was conducted to investigate the change in quality attributes of red pepper (paprika) (Capsicum annuum L. var. Km-622) as a function of ripening and some technological factors. Of quality attributes, carotenoids and bioantioxidants (ascorbic acid and tocopherols) have been studied. It was found that the dynamics of fruit ripening with regard to carotenoids and bioantioxidants was influenced to a considerable extent by weather conditions of the production season. A rainy and cool season yielded fruits with more beta-carotene but less diesters of red xanthophylls as compared to those produced in a relatively dry and warm season. The ripening stage at harvest was found to affect the quality of paprika. Harvest at unripe stages (color break or faint red) resulted in a high accumulation of dehydroascorbic acid in the overripe fruits, whereas de novo biosynthesis of carotenoids and tocopherols was partially retarded. Application of pre-drying centrifugation resulted in a marked loss of ascorbic acid, and as a consequence, carotenoid stability was impaired during the storage of ground paprika. Sugar caramelization caused dry pods and ground paprika to retain more pigments and tocopherol as compared to those from control or centrifuged red pepper samples. During the storage of ground paprika, color stability was improved by grinding the seeds with the pericarp.


[Morphological and cytophotometric characteristics of polyps of the colon and rectum].
Material from 60 patients with colon and rectum polyps as well as "normal" mucous membrane (44 patients) taken at some distance from polyps are studied. DNA content was measured by a flow cytometry method (ICP-II) in parallel to morphological (cytological and histological) examination of biopsies. Analysis of material was performed by groups depending on the epithelial dysplasia degree in the adenomas. Diploid cells dominated in the adenomas. Significant increase of cells in S- and (G2 + M)-phases of cell cycles is noted in the adenomas with a severe epithelial dysplasia as compared to the adenomas with a mild or moderate dysplasia. Aneuploid cells are detected in 5 adenomas, 4 of them morphologically were those with a severe dysplasia including 2 cases in which it was difficult to differentiate between a severe dysplasia and carcinoma although the signs of invasion were absent.


Changing pattern of histological type, location, stage and outcome of surgical treatment of gastric carcinoma.
There are indications that some features of gastric carcinoma are changing, with a possible impact on prognosis. The aim of this study was to examine any changes in type, location, stage, resection rate, postoperative mortality rate or prognosis for patients with gastric carcinoma in a well defined population. During 1974-1991, 1161 new cases of gastric adenocarcinoma were diagnosed in Ostergötland County, Sweden. Tumour location, Laurén histological type, tumour node metastasis (TNM) stage, radicality of tumour resection and postoperative complications were recorded after histological re-evaluation of tissue specimens and examination of all patient records. Dates of death were obtained from the Swedish Central Bureau of Statistics. Time trends were studied by comparing the intervals 1974-1982 (period 1) and 1983-1991 (period 2). The proportion of diffuse type of adenocarcinoma increased (from 27 to 35 per cent), while that of mixed type decreased (from 16 to 9 per cent) and that of intestinal type was unchanged. The proportion of tumours located in the proximal two-thirds of the stomach increased (from 32 to 42 per cent) and the proportion of patients with tumours in TNM stage IV decreased (from 32 to 25 per cent). Overall tumour resection rates were unchanged, although the proportion of radical total gastrectomies increased (from 36 to 50 per cent). Excluding tumours of the cardia or gastric remnant after previous ulcer surgery, the 5-year relative survival rate after radical resection increased from 25 to 36 per cent and the postoperative mortality rate decreased for both radical (from 11 to 4 per cent) and palliative (from 18 to 6 per cent) resection. The patterns of tumour histology, location and stage of gastric carcinoma have changed in the authors' region. These changes were paralleled by a significant improvement in survival and postoperative mortality rates.


Chemotherapy for schistosomiasis in Ugandan fishermen: treatment can cause a rapid increase in interleukin-5 levels in plasma but decreased levels of eosinophilia and worm-specific immunoglobulin E.
Chemotherapy for blood-dwelling schistosomes kills the worms and exposes parasite antigen to the circulation. In many people from areas of endemicity, this treatment increases parasite-specific immunoglobulin E (IgE) and other Th2 responses in the months following therapy, responses that have been associated with subsequent resistance to reinfection. Here we investigate much earlier changes in immune reactions after praziquantel therapy in Schistosoma mansoni-infected fishermen working in an area of high transmission in Uganda. The subjects gave blood before treatment and at 1 and 21 days posttreatment. Blood cultures were incubated with schistosome soluble worm antigen (SWA) or soluble egg antigen (SEA). Interleukin-4 (IL-4), IL-5, IL-10, IL-13, gamma interferon, and transforming growth factor beta levels were measured in the cultures and in plasma. A marked transient increase in plasma IL-5 levels was observed in 75% of the subjects (n = 48) by 1 day posttreatment. This response was dependent on pretreatment intensity of infection and was accompanied by a transient decrease in eosinophil numbers. One day posttreatment, blood cultures from the 16 subjects with the greatest increase in plasma IL-5 level (>100 pg/ml) displayed reduced IL-5, IL-13, and IL-10 responses to SWA, and in contrast to the rest of the cohort, these high-IL-5 subjects displayed reduced levels of SWA-specific IgE in plasma 21 days posttreatment. Twenty months after treatment, the intensity of reinfection was positively correlated with the increase in plasma IL-5 level seen 1 day posttreatment. These studies describe the heterogeneity in early immune reactions to treatment, identifying subgroups who have different patterns of reaction and who may have different capacities to mount the responses that have been associated with resistance to reinfection.


Levodopa does not change cerebral vasoreactivity in Parkinson's disease.
The aim of this work was to study cerebral vasoreactivity to hypercapnia in Parkinson's disease (PD) before and after levodopa administration. The prospective study was conducted in 20 patients presenting with PD, using 3T blood oxygenation level-dependent (BOLD) functional MRI (fMRI) covering the whole brain. The hypercapnic stimulus was block-designed using carbogen inhalation, a gas mixture of 7% CO2 and 93% O2, before (OFF) and 60 minutes after administration of a suprathreshold (120%) therapeutic L-dopa dose (ON). Ten age-matched controls were enrolled for between-group comparisons. Analyses were conducted with a random effects model and corrected for multiple comparisons. No adverse reaction to the hypercapnic stimulus was reported. However, 10 patients and 2 controls were excluded because of incomplete protocol realization, inappropriate hypercapnic stimulus, or excessive movements, leaving 10 patients and 8 controls for further analyses. The hypercapnic stimulus increased whole-brain BOLD signal of 1.48% ± 0.06% (mean ± standard error) in controls, 1.59% ± 0.05% in patients OFF, and 1.62% ± 0.09% in patients ON. Regions of interest analyses showed a signal increase in gray matter of 2.60% ± 0.16% in controls, 2.89% ± 0.21% in patients OFF, and 2.87% ± 0.12% in patients ON. No global or regional significant difference was detected, when comparing patients OFF and ON L-dopa, or between patients and controls. Contrary to Alzheimer's disease, the vasoreactivity to hypercapnia was normal in PD before and after L-dopa administration, compared to controls. This negative result is an important finding, especially for neuroscientists using fMRI to investigate motricity and cognition, discarding a significant confounding effect.


Divergent COVID-19 Disease Trajectories Predicted by a DAMP-Centered Immune Network Model.
COVID-19 presentations range from mild to moderate through severe disease but also manifest with persistent illness or viral recrudescence. We hypothesized that the spectrum of COVID-19 disease manifestations was a consequence of SARS-CoV-2-mediated delay in the pathogen-associated molecular pattern (PAMP) response, including dampened type I interferon signaling, thereby shifting the balance of the immune response to be dominated by damage-associated molecular pattern (DAMP) signaling. To test the hypothesis, we constructed a parsimonious mechanistic mathematical model. After calibration of the model for initial viral load and then by varying a few key parameters, we show that the core model generates four distinct viral load, immune response and associated disease trajectories termed "patient archetypes", whose temporal dynamics are reflected in clinical data from hospitalized COVID-19 patients. The model also accounts for responses to corticosteroid therapy and predicts that vaccine-induced neutralizing antibodies and cellular memory will be protective, including from severe COVID-19 disease. This generalizable modeling framework could be used to analyze protective and pathogenic immune responses to diverse viral infections.


The role of intact oleosin for stabilization and function of oleosomes.
Lipid storage in plants is achieved among all plant species by formation of oleosomes, enclosing oil (triacylglycerides) in small subcellular droplets. Seeds are rich in this pre-emulsified oil to provide a sufficient energy reservoir for growing. The triacylglyceride core of the oleosomes is surrounded by a phospholipid monolayer containing densely packed proteins called oleosins. They are anchored in the triacylglycerides core with a hydrophobic domain, while the hydrophilic termini remain on the surface. These specialized proteins are expressed during seed development and maturation. Particularly, they play a major role in the stabilization and function of oleosomes. To better understand the importance of oleosins for oleosome stabilization, enzymatic digestion of oleosins was performed. This made it possible to compare and correlate changes in the molecular structure of oleosins and changing macroscopic properties of oleosomes. Tryptic digestion cleaves the hydrophilic part of the oleosins, which is accompanied by a loss of secondary structures as evidenced by Fourier-transform infrared and sum frequency generation spectra. After digestion, the ability of oleosins to stabilize oil-water or air-water interfaces was lost. The surface charge and the associated aggregation behavior of oleosomes are governed by interactions typical of proteins before digestion and by interactions typical of phospholipids after digestion.


To investigate the prevention of OM-85 on bronchiectasis exacerbations (iPROBE) in Chinese patients: study protocol for a randomized controlled trial.
Non-cystic fibrosis bronchiectasis is characterized by the irreversible dilatation of the medium-sized bronchi as a result of airway injury from recurrent or chronic inflammation and lower respiratory tract infections. Bronchiectasis airways are commonly colonized with bacterial species. Infections of the airways play important role in bronchiectasis exacerbations. The non-specific prevention of recurrent airway infections by immunostimulating agents has gained growing interest. OM-85, consisting of extracts of eight kinds of bacteria important in respiratory infections, could support the respiratory tract resistance to the pathogens. OM-85 has been shown to be a benefit by decreasing the risk of acute exacerbation of chronic obstructive pulmonary disease (COPD) in several perspective clinical trials. Exacerbation of bronchiectasis substantially contributes to a more rapid decline in lung function, reduced quality of life, and healthcare costs. In this context, we plan to conduct a clinical trial to investigate the PReventive effect of OM-85 on Bronchiectasis Exacerbation in Chinese patients (iPROBE). This study is designed as a prospective, randomized, double blind, placebo-controlled multicenter trial. A total of 244 patients with bronchiectasis, who have had at least one exacerbation of bronchiectasis in the previous year, will be included. The subjects will randomly receive two courses of 7 mg of OM-85 or a matching placebo. The treatment dose of OM-85 will be one daily capsule taken orally for 10 days each month for 3 consecutive months at the beginning of the study, followed by 3 months of no drug. This schedule will repeat until the patient has been seen for one year. We will investigate whether long-term treatment with an oral immunostimulant (OM-85) could decrease exacerbations of bronchiectasis over a one-year period. We will also assess other relevant outcomes, including the rate of event-based exacerbation, lung function parameters, and total scores judged by the St George's respiratory questionnaire, Leicester cough questionnaire, and inflammatory index. We hope that this study will provide new information on the preventive effects of OM-85 on bronchiectasis exacerbations and will address a knowledge gap for this understudied disease. This study is registered at http://www.clinicaltrials.gov (identifier NCT01968421) on 19 October 2013.


Retrospective review of osteoarticular infections in a pediatric sickle cell age group.
Patients with sickle cell disease have been documented to be particularly susceptible to osteoarticular infections. Controversy exists concerning the bacteriology, etiology, and clinical presentation in differentiating osteoarticular infections from bone infarct. We retrospectively reviewed all cases from our institution over the past 22 years of osteoarticular infections in children who carry the diagnosis of sickle cell disease. Two thousand consecutive patient charts of children enrolled in the Pediatric Sickle Cell Clinic of our institution between 1973 and 1995 were evaluated. There were 14 cases of bone or joint infections (10 osteomyelitis, four septic arthritis). There was one case of multicentric osteomyelitis and one case of meningitis complicating the septic arthritis. There were nine male and five female patients with ages ranging from 6 months to 17 years (mean, 8.0). All patients were noted to have hemoglobin SS. The predominant presenting symptoms were pain (79% of cases) and swelling (71% of cases). The most frequent physical findings were fever >38.2 degrees C (71% of cases) and tenderness (86% of cases). Ninety-three percent of the children had a white blood count exceeding 15,000/mm3 (range, 7,900-32,300). Westergren sedimentation rates ranged from 14 to 89 mm/h with 93% of the children exceeding the normal value in our hospital. Cultures were positive in 75% of tissue biopsies, 58% of the blood cultures, and 70% of the bone or joint aspirates. The most common offending organism found in osteomyelitis was Salmonella (eight of 10 cases); however, no predominant organism found was identified in cases of septic arthritis. Radiographs and bone scans were of limited value in the differential diagnosis between osteoarticular infections and bone infarction. Early diagnosis and treatment of osteoarticular infections is key to satisfactory outcome. This study suggests that an ill-appearing patient with a fever >38.2 degrees C, pain, and swelling should prompt the physician to aspirate or biopsy the area and not rely on diagnostic studies that we found to be unreliable.


Optimisation of shock wave lithotripsy: a systematic review of technical aspects to improve outcomes.
Shock wave lithotripsy (SWL) remains an important treatment option for the management of upper urinary tract stones. The optimisation of certain technical principles can help to improve the results of SWL. We performed a systematic review based on preferred reporting items for systematic review and meta-analysis (PRISMA) standards for studies reporting on technical aspects of SWL. A literature search was conducted on the PubMed database between January 1984 and November 2018 using 'shockwave lithotripsy' and 'stone' as keywords. Summaries and manuscripts of relevant articles were reviewed in order to select studies with the best level of evidence in each theme covered during the review. From 4,135 titles, 165 abstracts and full-text articles were reviewed. Overall, SWL has good outcomes in the treatment of upper urinary tract stones. It remains the only truly non-invasive stone treatment. While stone-free rate (SFR) might not be equivalent to ureteroscopy or percutaneous nephrolithotomy outcomes, SWL can be optimised by changing several technical factors, including type of machine, patient position, number, rate and energy of shocks, stone targeting, and patient analgesia. For each of these included SWL themes, relevant and selected studies with the highest level of evidence were described and discussed. Paired with these improved technical factors and appropriate patient selection, SWL, with its low complication rates, remains an excellent treatment option in 2019.


Commentary: sharper instruments: on defending the humanities in undergraduate medical education.
The study by Ousager and Johannessen in this issue finds a lack of research attempting to measure the long-term effects of incorporating humanities into the undergraduate medical education (UME) curriculum, and warns that more such studies are needed if the humanities are to become integrated into UME. This commentary points to limitations in the study's methodology, suggesting that the value of the humanities in educating new physicians can be defended by demonstrating the need for more complex approaches to knowledge than complete dependence on empirical evidence, and invites those who support inclusion of the humanities in UME to take up three challenges: work together to define the terms and scope of the medical humanities as a coherent (though heterogenous) field, teach reading skills (promote, that is, a nonreductive approach to the interpretation of human objects), and work to establish effective and persuasive alternatives to the blunt tools of outcomes measurement.


A simple microviscometric approach based on Brownian motion tracking.
Viscosity-an integral property of a liquid-is traditionally determined by mechanical instruments. The most pronounced disadvantage of such an approach is the requirement of a large sample volume, which poses a serious obstacle, particularly in biology and biophysics when working with limited samples. Scaling down the required volume by means of microviscometry based on tracking the Brownian motion of particles can provide a reasonable alternative. In this paper, we report a simple microviscometric approach which can be conducted with common laboratory equipment. The core of this approach consists in a freely available standalone script to process particle trajectory data based on a Newtonian model. In our study, this setup allowed the sample to be scaled down to 10 μl. The utility of the approach was demonstrated using model solutions of glycerine, hyaluronate, and mouse blood plasma. Therefore, this microviscometric approach based on a newly developed freely available script can be suggested for determination of the viscosity of small biological samples (e.g., body fluids).


A review on the bioavailability, bio-efficacies and novel delivery systems for piperine.
As the major naturally occurring alkaloid in pepper with a pungent taste, piperine is known for its beneficial biological functions and therapeutic effects. In this work, the bioavailability and biological activities of piperine were presented and discussed. Novel delivery systems for enhancing the bioavailability of piperine were also reviewed. This study could provide a better understanding of the physiological and biochemical aspects of piperine to be further developed in the food and nutraceutical industries.


In vivo transmigrated monocytes from patients with stable coronary artery disease have a reduced expression of CD11b.
Coronary artery disease (CAD) is characterized by infiltration of monocyte derived cells in the intima of the vessel wall. We hypothesized that accumulation of these cells is caused partly by an altered monocyte transmigration process in CAD. To gain insight into this issue we applied the skin blister method that allows collection of in vivo transmigrated cells at sites of local inflammation. Nineteen patients with stable CAD and 19 matched controls were enrolled. Markers of inflammation and gradients of chemokines, as well as adhesion molecule expression and up-regulation capacity, were studied. The expression of inflammatory markers, such as C-reactive protein, interleukin (IL)-6, tumour necrosis factor-alpha and IL-10, was similar in patients and controls, indicating that patients were in a stable phase of the disease. Expression of adhesion molecules, CD11b and very late activation antigen-4, on peripheral monocytes did not differ between patients and controls. However, following in vivo transmigration, monocytes in patients with CAD had a significantly reduced expression and mobilization of CD11b. The effect on CD11b could not be reproduced by in vitro stimulation with blister fluid, representing a local inflammatory milieu, or in an in vitro system of transmigration. These findings point towards differences in monocyte CD11b expression and availability at an inflammatory site between patients with CAD and healthy controls.


In situ fluorescent protein imaging with metal film-enhanced total internal reflection microscopy.
Fluorescence detection of single molecules provides a means to investigate protein dynamics minus ambiguities introduced by ensemble averages of unsynchronized protein movement or of protein movement mimicking a local symmetry. For proteins in a biological assembly, taking advantage of the single molecule approach could require single protein isolation from within a high protein concentration milieu. Myosin cross-bridges in a muscle fiber are proteins attaining concentrations of approximately 120 muM, implying single myosin detection volume for this biological assembly is approximately 1 attoL (10(-18) L) provided that just 2% of the cross-bridges are fluorescently labeled. With total internal reflection microscopy (TIRM) an exponentially decaying electromagnetic field established on the surface of a glass-substrate/aqueous-sample interface defines a subdiffraction limit penetration depth into the sample that, when combined with confocal microscopy, permits image formation from approximately 3 attoL volumes. Demonstrated here is a variation of TIRM incorporating a nanometer scale metal film into the substrate/glass interface. Comparison of TIRM images from rhodamine-labeled cross-bridges in muscle fibers contacting simultaneously the bare glass and metal-coated interface show the metal film noticeably reduces both background fluorescence and the depth into the sample from which fluorescence is detected. High contrast metal film-enhanced TIRM images allow secondary label visualization in the muscle fibers, facilitating elucidation of Z-disk structure. Reduction of both background fluorescence and detection depth will enhance TIRM's usefulness for single molecule isolation within biological assemblies.


Isolation and spectral characterization of photochemical reaction centers from the thermophilic green bacterium Chloroflexus aurantiacus strain J-10-f1.
A rapid procedure has been devised to extract photochemically active reaction centers from the green bacterium Chloroflexus aurantiacus strain J-10-f1 and a mutant lacking colored carotenoids (73-3). The isolated material was completely free of antenna bacteriochlorophyll and cytochromes and nearly free of carotenoids and had near IR absorption maxima at 865, 815, and 756 nm. On oxidation, the peak at 865 nm was bleached and the remaining two peaks were shifted to 806 and 757 nm. Although these spectral characteristics show general similarities to those in reaction center preparations from purple bacteria, there are some distinct differences. Comparison of the spectra of reaction centers of Chloroflexus and Rhodopseudomonas sphaeroides, adjusted to the same absorbance at 865 nm, showed that the absorbance at 815 nm in Chloroflexus was much less and that at 757 nm was much greater than the equivalent absorbances in R. sphaeroides. Unlike reaction centers from R. sphaeroides and other photosynthetic bacteria that have two molecules of bacteriopheophytin and four molecules of bacteriochlorophyll per unit, reaction centers from Chloroflexus appear to have three molecules of each pigment per unit. A prominent shoulder at 793 nm disappears concomitantly with the bleaching at 865 nm on oxidation of Chloroflexus reaction centers; this spectral component may represent the higher energy transition in the near IR of the two bacteriochlorophylls forming P865. While Chloroflexus has a light-harvesting pigment system very similar to that of the Chlorobiaceae, its reaction center has optical absorption characteristics similar to those of the Rhodospirillaceae.


Depressive Symptoms in Autistic Youth with Anxiety Disorders.
Anxiety and depression often coexist in youth and share overlapping symptomatology; however, little is known about the comorbidity of anxiety and depression in autistic youth. This study explores (1) the frequency of depressive symptoms among autistic children with clinically significant anxiety, (2) clinical variables that may be associated with elevated depressive symptoms, and (3) whether pretreatment depressive symptoms predict cognitive behavioral therapy (CBT) outcomes for anxiety. Children aged 7 to 13 years (N = 87) and their parents participated in a randomized controlled trial comparing 2 versions of a parent-led, telehealth-delivered CBT program. Parents and children completed a variety of clinical assessments and self-report questionnaires before and after treatment. Fifty-seven percent of the child sample reported experiencing elevated depressive symptoms while roughly 20% of parents reported elevated depressive symptoms in their child. A strong association between anxiety and depression was found. Heightened feelings of loneliness, per child report, and functional impairment, per parent report, were found to be uniquely associated with elevated depressive symptoms. Finally, depressive symptoms were not a significant predictor of CBT outcomes for anxiety. Findings suggest high degrees of comorbidity between anxiety and depression among autistic children and that feelings of loneliness, anxiety, and functional impairment may be early indicators of mood-related concerns. Further research is needed to determine the full extent of the association between anxiety and depression and additional options for treating depression in autistic children.


Diaphorase Coupling Protocols for Red-Shifting Dehydrogenase Assays.
Dehydrogenases are an important target for the development of cancer therapeutics. Dehydrogenases either produce or consume NAD(P)H, which is fluorescent but at a wavelength where many compounds found in chemical libraries are also fluorescent. By coupling dehydrogenases to diaphorase, which utilizes NAD(P)H to produce the fluorescent molecule resorufin from resazurin, the assay can be red-shifted into a spectral region that reduces interference from compound libraries. Dehydrogenases that produce NAD(P)H, such as isocitrate dehydrogenase 1 (IDH1), can be read in kinetic mode. Dehydrogenases that consume NAD(P)H, such as mutant IDH1 R132H, can be read in endpoint mode. Here, we report protocols for robust and miniaturized 1,536-well assays for WT IDH1 and IDH1 R132H coupled to diaphorase, and the counterassays used to further detect compound interference with the coupling reagents. This coupling technique is applicable to dehydrogenases that either produce or consume NAD(P)H, and the examples provided here can act as guidelines for the development of high-throughput screens against this enzyme class.


Influence of genes encoding proton-translocating enzymes on suppression of Salmonella typhimurium growth and colonization.
Twenty-four-hour-old, aerobically grown, Luria-Bertani broth cultures of Salmonella typhimurium F98 suppressed the growth of a spectinomycin-resistant (Spcr) derivative of the same strain inoculated at 10(3) CFU ml(-1). This growth suppression is genus specific and RpoS independent, and it is not solely a result of nutrient depletion (P. A. Barrow, M. A. Lovell, and L. Zhang-Barber, J. Bacteriol. 178:3072-3076, 1996). Mutations in three genes are shown here to significantly reduce growth suppression under these conditions. The mutations were located in the nuo, cyd, and unc operons, which code for the NADH dehydrogenase I, cytochrome d oxidase, and F0F1 proton-translocating ATPase complexes, respectively. When cultures were grown under strictly anaerobic conditions, only the unc mutant did not suppress growth. Prior colonization of the alimentary tract of newly hatched chickens with the S. typhimurium F98 wild type or nuo or cyd mutants suppressed colonization by an S. typhimurium F98 Spcr derivative inoculated 24 h later. In contrast, the S. typhimurium unc mutant did not suppress colonization. The nuo and unc mutants showed poorer growth on certain carbon sources. The data support the hypothesis that growth suppression operates because of the absence of a utilizable carbon source or electron acceptor.


The social environment of schools and adolescent nutrition: associations between the school nutrition climate and adolescents' eating behaviors and body mass index.
The aim of this study was to determine the association between the school nutrition climate and students' eating behaviors and body mass index (BMI). Data were collected as part of Youth'07, a nationally representative health survey of high school students in New Zealand. Overall, 9107 randomly selected students from 96 randomly selected schools participated. School-level measures were created by aggregating students' reports within schools. Analyses were conducted using multilevel modeling, accounting for student-level characteristics. There was a positive association between the school nutrition climate and students' consumption of fruits and vegetables. This relationship was statistically significant after controlling for the background characteristics of students. There were no associations between the school nutrition climate and students' junk food consumption or BMI. The school nutrition climate appears to have a positive influence on adolescents' healthy eating behaviors (fruit and vegetable intake), but a limited effect on unhealthy eating behaviors and ultimately body weight. This may reflect the pervasiveness of junk food in the environments of adolescents outside of school and the difficulty in limiting its consumption.


Dopaminergic Genetic Variation in Young Adolescents: Associations with Sensation-Seeking.
Deficient reward functioning, including reward-related personality, is implicated in depression's etiology. A dopaminergic genetic multilocus genetic profile score (MGPS) has previously been associated with neural reward responsivity but, despite theoretical basis, has not been studied with reward-related personality. Such research is needed to elucidate associations between genetic variation and reward-related personality in a developmentally sensitive population. In the present study, we examined associations between dopaminergic MGPS's and self-report reward-related personality in two young adolescent samples aged 10-15 years old (Sample 1: N = 100 girls, 82% White, 18% Other; Sample 2: N = 141, 65 girls, 76 boys, 89.36% White, 10.64% Other) using an established MGPS and an augmented MGPS. A "mini" meta-analysis synthesized results across samples. In Sample 1, an exploratory mediation analysis intended to gauge effect size for future work tested a path between the MGPS and depression through significant reward traits. In each independent sample, both MGPS's showed significant associations with sensation-seeking but not social drive, a pattern that persisted following correction. Effect sizes of novel variants were at least as robust as established variants, suggesting their added utility. Additionally, the exploratory mediation analysis suggested no noteworthy indirect effect, but a small (R2 = 0.022), statistically non-significant direct effect of the MGPS predicting prospective depressive symptoms. Results suggest that dopaminergic genetic variation is associated with the reward-related personality trait of sensation seeking but not social drive. Additional work is needed to probe whether sensation seeking may be a path through which this genetic variation confers depression risk.


Identification of proliferin mRNA and protein in mouse placenta.
Proliferin is a recently described, prolactin-related protein whose mRNA appears in several murine cell lines during active growth. We have surveyed a number of mouse organs or tissues for the presence of mRNAs that hybridize to cloned proliferin cDNA. Of the tissues tested, only the placenta yielded proliferin-related mRNA. This placental RNA is about 1 kilobase in length, increases sharply between days 8 and 10 of pregnancy, and then gradually declines through day 18. It is more abundant in RNA extracted from the fetal, compared to the maternal, part of the placenta. From a cDNA plasmid library prepared from poly(A)+ placental RNA, two types of proliferin-related clones were isolated, differing in intensity of hybridization to proliferin cDNA. By nucleotide sequence analysis, a strongly hybridizing clone was found to be nearly identical to the proliferin cDNA clone isolated from a library prepared from mRNA of a growing mouse fibroblastic cell line. Using an antiserum prepared against a synthetic proliferin fusion protein, we show that proliferin is secreted as a glycoprotein by minced placental tissue and that it differs from mouse placental lactogen. We conclude that proliferin is a placental hormone that is synthesized in certain mouse cell lines during active growth. Its function during pregnancy and during the growth of cultured cells is presently unknown.