Vitamin E-stabilized UHMWPE: Biological response on human osteoblasts to wear debris.
UHMWPE doped with vitamin E was introduced to provide oxidation resistance upon sterilization, without affecting UHMWPE's mechanical properties. Particle-induced macrophage activation leads to periprosthetic bone resorption, requiring total joint replacements. During osteolysis, osteoblasts produce osteoimmunological factors such as RANKL and OPG, and the inhibitors of the Wnt pathway DKK-1 and Sclerostin. This study investigated in vitro how vitamin E-blended-UHMWPE wear debris might affect osteoblast-mediated osteolysis and the production of RANKL, OPG, Sclerostin and DKK-1, compared to conventional UHMWPE wear debris. Human osteoblastic SaOS2 cells were incubated with wear particles from Vitamin E doped and conventional UHMWPE and the gene expression and protein production of IL-6, RANKL, OPG, DKK-1, and Sclerostin was evaluated, RANKL, a bone erosion marker, was reduced, while OPG, a bone protective marker, were increased by the vitamin E-blended UHMWPE compared to conventional UHMWPE. Vitamin E doped UHMWPE reduced Sclerostin level, and partially affected DKK-1 production, thereby protecting against bone erosion. In conclusion, Vitamin E-blended UHMWPE induced an osteoimmunological response in bone cells that had positive effects on the osteolysis induced by wear debris, reducing aseptic loosening of the implants. In conclusion, this is the first study showing that Vitamin E-blended UHMWPE induced an osteoimmunological response in bone cells that positively affect the osteolysis induced by wear debris, thereby reducing the aseptic loosening of the implants.


Corticotropin/lipotropin common precursor-like material in normal rat extrapituitary tissues.
A Mr 26,000 corticotropin (ACTH)-like material is present in glacial acetic acid extracts of all normal rat extrapituitary tissues. In the present study, beta-melanotropin (beta-MSH) immunoactivity was detected in glacial acetic acid extracts of normal rat extrapituitary tissues. beta-MSH immunoactivity was also present in all extracts (mean +/- SEM, fmol/mg of protein): brain, 71.0 +/- 16.3; stomach, 11.5 +/- 1.6; kidney, 8.9 +/- 0.8; colon, 8.2 +/- 1.1; small intestine, 6.5 +/- 1.1; liver, 4.3 +/- 0.5; and heart, 3.2 +/- 0.5. Except in brain extracts, beta-MSH and ACTH immunoactivities of tissue extracts were strongly correlated to each other (r = 0.79; n = 42). When tissue extracts (except brain) were passed through a Sephadex G-75 (superfine) column, ACTH and beta-MSH immunoactivities were eluted in a single peak corresponding to Mr 26,000. In contrast, for brain extracts, the MrS of major peaks of ACTH and beta-MSH immunoactivities were 4,500 and 8,000, respectively; a smaller peak of Mr 26,000 ACTH/beta-MSH-like material was also eluted. Specific anti-ACTH immunocolumns, which did not bind purified synthetic beta-MSH, adsorbed both ACTH and beta-MSH immunoactivities of all tissue extracts except those of brain. One-third of the beta-MSH immunoactivity in brain extracts adsorbed to the anti-ACTH immunocolumn, but two-thirds of beta-MSH immunoactivity passed through the column. We conclude that ACTH and beta-MSH immunoactivities are present in all normal rat extrapituitary tissues and exist in most tissues on the same molecule. This Mr 26,000 molecule is closely related to the pituitary ACTH/beta-lipotropin common precursor.


Coronavirus Disease (COVID-19): The Value of Chest Radiography for Patients Greater Than Age 50 Years at an Earlier Timepoint of Symptoms Compared With Younger Patients.
Background: A relative paucity of data exists regarding chest radiography (CXR) in diagnosis of coronavirus disease (COVID-19) compared to computed tomography. We address the use of a strict pattern of CXR findings for COVID-19 diagnosis, specifically during early onset of symptoms with respect to patient age. Methods: We performed a retrospective study of patients under investigation for COVID-19 who presented to the emergency department during the COVID-19 outbreak of 2020 and had CXR within 1 week of symptoms. Only reverse transcription polymerase chain reaction (RT-PCR)-positive patients were included. Two board-certified radiologists, blinded to RT-PCR results, assessed 60 CXRs in consensus and assigned 1 of 3 patterns: characteristic, atypical, or negative. Atypical patterns were subdivided into more suspicious or less suspicious for COVID-19. Results: Sixty patients were included: 30 patients aged 52 to 88 years and 30 patients aged 19 to 48 years. Ninety-three percent of the older group demonstrated an abnormal CXR and were more likely to have characteristic and atypical-more suspicious findings in the first week after symptom onset than the younger group. The relationship between age and CXR findings was statistically significant (χ2 [2, n=60]=15.70; P=0.00039). The relationship between negative and characteristic COVID-19 CXR findings between the 2 age cohorts was statistically significant with Fisher exact test resulting in a P value of 0.001. Conclusion: COVID-19 positive patients >50 years show earlier, characteristic patterns of statistically significant CXR changes than younger patients, suggesting that CXR is useful in the early diagnosis of infection. CXR can be useful in early diagnosis of COVID-19 in patients older than 50 years.


Insulin resistance in critically ill patients with acute renal failure.
Mortality in critically ill patients with acute renal failure (ARF) remains high. Hyperglycemia associated with insulin resistance has been associated with adverse outcomes in critically ill intensive care unit (ICU) patients but has not been examined specifically in patients with ARF. We used data from a subcohort (n = 90) of the Program to Improve Care in Acute Renal Disease (PICARD), an observational study of 618 adult ICU patients with ARF in whom nephrology service consultation was obtained. We obtained simultaneous measurements of serum glucose, insulin, insulin-like growth factor (IGF)-I, and IGF-1 binding proteins (IGFBP) in 90 patients. Daily glucose determinations were obtained from a larger fraction of the PICARD cohort (n = 509). Among the 90 patients with intensive metabolic monitoring, glucose concentrations in survivors were significantly lower than in nonsurvivors throughout the 5-wk period (P = 0.008, adjusted P = 0.013). In the larger PICARD cohort (n = 509), hyperglycemia was also significantly associated with in-hospital mortality. Mean insulin concentrations were significantly higher (431 +/- 508 vs. 234 +/- 189 pmol/l, P = 0.03), mean homeostasis model of insulin resistance levels were significantly higher (24.1 +/- 30.0 vs. 11.7 +/- 12.5, P = 0.04), and IGFBP-3 concentrations were significantly lower (1,190 +/- 498 vs. 1,470 +/- 581 microg/l, P = 0.02) among nonsurvivors compared with survivors. Insulin resistance as defined by hyperglycemia in the setting of higher insulin concentrations may be associated with mortality in critically ill patients with ARF. The IGF-IGFBP axis may play an important role in this process.


The Human-Nature Experience: A Phenomenological-Psychoanalytic Perspective.
Drawing upon phenomenology and psychoanalytic concepts, we explore and explicate participants' lived experience of the natural world. The authors draw upon Husserl's description of consciousness as intentionality and his later work on the life-world, in exploring experiences which provide a basis for a psychochoanalytic understanding of the human-nature experience. Unstructured interviews were undertaken with nine participants, each of whom regarded nature as being significant for their sense of wellbeing. The lived experiences were explicated drawing upon the two processes: Giorgi's descriptive phenomenological psychological methodology and psychoanalytic researcher reflexivity. Data analysis and explication involved the following steps: (1) a thorough reading of each interview transcript, (2) breaking data into parts by demarcating meaning units, (3) organizing data by translating meaning units into units of psychological experience through coding, and (4) arriving at a summary of the data which involved organizing and reviewing units of psychological experience. The process of reflection led to the formulation of an essential psychological structure of participants' lived experience of the natural world. We argue that the human-nature relationship can be conceived in terms of psychoanalytic concepts, and in particular, constructs based upon an understanding of the primacy of attachment relationships. The natural world is elucidated as (a) nature being experienced as a primary attachment, (b) nature experienced as a secure base, (c) nature experienced as twinship, (d) nature experienced as containing, and (e) nature experienced as embodied. This paper extends previous empirical descriptions of the human-nature relationship by incorporating psychoanalytic processes and theory into a theoretically informed qualitative methodological stance. Beyond the traditional notion of nature as something 'out there' that we can interact with for cognitive or emotional restoration, participants in this study described the experience of nature as being integral to their sense of self. This study suggests that experiences that facilitate immersion in nature provide opportunities for the development of an integrated sense of self that has a profound impact on a participant's sense of wellbeing. The findings further demonstrate the convergence between phenomenology and psychoanalytic constructs which offers a richness to our understanding the subjectivity of participants and their relationship with nature, a perspective not often attainable through more traditional quantitative research methodologies.


Multimodality therapy for locoregional extrahepatic cholangiocarcinoma: a population-based analysis.
Although surgical resection is the mainstay of treatment for extrahepatic cholangiocarcinoma, the majority of patients present with advanced disease. Due in part to numeric rarity, the optimum role of radiotherapy (RT) for extrahepatic cholangiocarcinoma, as well as its relative benefit, is an area of debate. The specific aim of this series was to estimate survival for extrahepatic cholangiocarcinoma patients receiving surgery and adjuvant RT using a robust population-based data set. Data were extracted from the Surveillance, Epidemiology, and End Results (SEER) limited-use data set for selected extrahepatic cholangiocarcinoma cases. Lognormal multivariate survival analysis was implemented to estimate survival for patients for treatment cohorts based on extent of surgical intervention and RT. Parametric estimated median survival for patients receiving total/radical resection + RT was 26 months; it was 25 months for total/radical resection alone, 25 months for subtotal/debulking resection + RT, 21 months for subtotal/debulking resection, 12 months for RT alone, and 9 months for those not receiving surgery or RT. Parametric multivariate analysis revealed age, American Joint Committee on Cancer Stage, grade, and surgical/radiation regimen as statistically significant covariates with survival. Surgery alone and adjuvant RT cohorts demonstrated evidence of improved survival compared with no treatment; comparatively, RT alone was associated with survival decrement. Early improvement in survival in adjuvant cohorts was not observed at later time points. Survival estimates using SEER data suggest an early survival advantage for adjuvant RT for patients with locoregional extrahepatic cholangiocarcinoma. Although future prospective series are needed to confirm these observations, SEER data represent the largest domestic population-based extrahepatic cholangiocarcinoma cohort, and may provide useful baseline survival estimates for future studies.


Measures, markers, and mediators: toward a staging system for clinical sepsis. A report of the Fifth Toronto Sepsis Roundtable, Toronto, Ontario, Canada, October 25-26, 2000.
Sepsis is not a single disease but a complex and heterogeneous process. Its expression is variable, and its severity is influenced by the nature of the infection, the genetic background of the patient, the time to clinical intervention, the supportive care provided by the clinician, and a number of factors as yet unknown. The evaluation of effective therapies has been hampered by limitations in our ability to characterize the process and to stratify patients into more homogeneous groups with respect to pathogenesis. To develop a taxonomy of markers relevant to clinical research in sepsis and to propose a testable candidate system for stratifying patients into more therapeutically homogeneous groups. An expert roundtable discussion and a MEDLINE review using search terms "marker" and "sepsis." Markers provide information in one or more of three domains: diagnosis, prognosis, and response to therapy. More than 80 putative markers of sepsis have been described. All correlate with the risk of mortality (prognosis), yet none has shown utility in stratifying patients with respect to therapy (diagnosis) or in titrating that therapy (response). Their limitations arise from the challenges of establishing causality in a complex disease process such as sepsis and of stratifying patients into more homogeneous populations. The former limitation may be addressed through a modification of Koch's postulates to differentiate causality from simple association. The latter suggests the need for a staging system analogous to those used in other complex disease processes such as cancer. A candidate framework for such a system, based on the infection, the host response, and the extent of organ dysfunction (the IRO system) is described. Advances in the understanding and management of patients with sepsis will necessitate more rigorous approaches to disease description and stratification. Models should be developed, tested, and modified through clinical studies rather than through consensus.


The shape of the spatial kernel and its implications for biological invasions in patchy environments.
Ecological and epidemiological invasions occur in a spatial context. We investigated how these processes correlate to the distance dependence of spread or dispersal between spatial entities such as habitat patches or epidemiological units. Distance dependence is described by a spatial kernel, characterized by its shape (kurtosis) and width (variance). We also developed a novel method to analyse and generate point-pattern landscapes based on spectral representation. This involves two measures: continuity, which is related to autocorrelation and contrast, which refers to variation in patch density. We also analysed some empirical data where our results are expected to have implications, namely distributions of trees (Quercus and Ulmus) and farms in Sweden. Through a simulation study, we found that kernel shape was not important for predicting the invasion speed in randomly distributed patches. However, the shape may be essential when the distribution of patches deviates from randomness, particularly when the contrast is high. We conclude that the speed of invasions depends on the spatial context and the effect of the spatial kernel is intertwined with the spatial structure. This implies substantial demands on the empirical data, because it requires knowledge of shape and width of the spatial kernel, and spatial structure.


Effects of Modern Food Retailers on Adult and Child Diets and Nutrition.
In many developing countries, food environments are changing rapidly, with modern retailers-such as supermarkets-gaining in importance. Previous studies have suggested that the rise of modern retailers contributes to overweight and obesity. Effects of modern retailers on dietary quality have not been analyzed previously due to the unavailability of individual-level dietary data. Here, we address this research gap with data from randomly selected households in Lusaka, Zambia. Anthropometric and food-intake data from 930 adults and 499 children were analyzed to estimate effects of purchasing food in modern retailers on body weight, height, and dietary quality while controlling for income and other confounding factors. The food expenditure share spent in modern retailers was found to be positively associated with overweight in adults, but not in children. For children, a positive association between expenditures in modern retailers and height was identified. Modern retailers contribute to higher consumption of ultra-processed foods and calories. But they also increase protein and micronutrient intakes among adults and children, mainly through higher consumption of meat and dairy. The findings underline that modern retailers can influence diets and nutrition in positive and negative ways. Differentiated regulatory policies are needed to shape food environments for healthy food choices and nutrition.


Substance use among adolescent high school students in India: A survey of knowledge, attitude, and opinion.
Is knowledge regarding the consequences of substance use among adolescents enough to prevent them from initiating and continuing its use, is a question that needs to be clarified further? To assess the knowledge regarding harm of use and to obtain information about attitudes among high school students. Also, to discover the opinion of substance use held by users. This was a population based cross-sectional study conducted in two high schools of West Bengal, India, among 416 students, in classes VIII, IX, and X, with no interventions. Primary outcome measurements were substance use: knowledge regarding harm, attitude, and opinion. Following this proportions and the chi-square test were used for statistical analysis. Out of 416 students, 52 (12.5%) used or abused any one of the substances irrespective of time and frequency in lifetime; 26 (15.1 %) were among the urban students and 26 (10.7 %) were among their rural counterparts. More than two-thirds (73.07%) of the respondents expressed a desire to quit substance use and 57.69% had tried to stop. 'Easy availability' and 'relief from tension' were the most frequent reasons for continuation of substance use. Level of knowledge on harmfulness of substance use among students was very high (urban - -84.6% and rural - 61.5%) and they stated media as the most frequent source of information. Users were successful in influencing their peers into taking up this habit (urban - 15.4% and rural - 26.9%). Inspite of being aware of the harmful effects of substance use, adolescents take up this habit. This requires comprehensive prevention and control programs in schools and the community, targeted toward adolescents and their parents and other family members. Effective measures are required to encourage shaping the attitude of school children toward self-confidence and adequacy, as also to prevent risk behavior among adolescents.


Risk-adjusted models for adverse obstetric outcomes and variation in risk-adjusted outcomes across hospitals.
Regulatory bodies and insurers evaluate hospital quality using obstetrical outcomes, however meaningful comparisons should take preexisting patient characteristics into account. Furthermore, if risk-adjusted outcomes are consistent within a hospital, fewer measures and resources would be needed to assess obstetrical quality. Our objective was to establish risk-adjusted models for 5 obstetric outcomes and assess hospital performance across these outcomes. We studied a cohort of 115,502 women and their neonates born in 25 hospitals in the United States from March 2008 through February 2011. Hospitals were ranked according to their unadjusted and risk-adjusted frequency of venous thromboembolism, postpartum hemorrhage, peripartum infection, severe perineal laceration, and a composite neonatal adverse outcome. Correlations between hospital risk-adjusted outcome frequencies were assessed. Venous thromboembolism occurred too infrequently (0.03%; 95% confidence interval [CI], 0.02-0.04%) for meaningful assessment. Other outcomes occurred frequently enough for assessment (postpartum hemorrhage, 2.29%; 95% CI, 2.20-2.38, peripartum infection, 5.06%; 95% CI, 4.93-5.19, severe perineal laceration at spontaneous vaginal delivery, 2.16%; 95% CI, 2.06-2.27, neonatal composite, 2.73%; 95% CI, 2.63-2.84). Although there was high concordance between unadjusted and adjusted hospital rankings, several individual hospitals had an adjusted rank that was substantially different (as much as 12 rank tiers) than their unadjusted rank. None of the correlations between hospital-adjusted outcome frequencies was significant. For example, the hospital with the lowest adjusted frequency of peripartum infection had the highest adjusted frequency of severe perineal laceration. Evaluations based on a single risk-adjusted outcome cannot be generalized to overall hospital obstetric performance.


Gastric bypass as treatment for obesity: trends, characteristics, and complications.
This paper describes national trends in gastric bypass procedures from 1998 through 2003 and explores the demographic and health profile of those who receive this procedure. Short-term outcomes such as length of stay and in-hospital complication rates are also examined. Data on obese hospital inpatients who had gastric bypass were obtained from the 1998 to 2003 National Hospital Discharge Survey. Gastric bypass was reported for an estimated 288,000 discharges during the 6-year study period. Trends within the 6-year period were tested using weighted regression. Characteristics of gastric bypass patients were compared with those of other inpatients using a chi(2) test of independence and the two-sided t test. The estimated number of hospital discharges with gastric bypass increased significantly, from 14,000 in 1998 to 108,000 in 2003. During this period, the average length of stay declined by 56% from 7.2 to 3.2 days. Gastric bypass patients were primarily women (84%), 25 to 54 years of age (82%), and privately insured (76%). A 1 in 10 complication rate was found for discharges with gastric bypass. Gastric bypass procedures in the United States have increased rapidly since 1998, whereas the average hospital stay has decreased. The decreasing length of stay needs to be evaluated in conjunction with potential complication rates and the permanent change in anatomy and lifestyle that must accompany this procedure. Monitoring trends in use of this procedure is important, especially if reimbursement policies change and the epidemic of obesity continues.


CT, MR and SPECT imaging in temporal lobe epilepsy.
Cranial computed tomography (CT) with modified temporal lobe technique, 0.15T magnetic resonance imaging (MRI) and single photon emission computed tomography (SPECT) were carried out on 30 patients with intractable temporal lobe epilepsy. Lateralising abnormalities were detected in 21/30 patients overall. Specific lesions were detected by CT in one patient and by MRI in seven patients (in one case bilateral). In addition CT detected asymmetry of the sylvian fissures or temporal horns in 10 patients, and MRI in eight patients. SPECT detected lateralising abnormalities in 19 patients (in five cases bilateral). It is concluded that low field MRI is superior to modified CT in demonstrating subtle structural lesions of the temporal lobe. Functional scanning with SPECT supports the evidence of origin of an epileptic focus in a substantial proportion of cases and may improve the selection of patients for surgery.


The great balancing act: regulation and fate of antiviral T-cell interactions.
The fate of T lymphocytes revolves around a continuous stream of interactions between the T-cell receptor (TCR) and peptide-major histocompatibility complex (MHC) molecules. Beginning in the thymus and continuing into the periphery, these interactions, refined by accessory molecules, direct the expansion, differentiation, and function of T-cell subsets. The cellular context of T-cell engagement with antigen-presenting cells, either in lymphoid or non-lymphoid tissues, plays an important role in determining how these cells respond to antigen encounters. CD8(+) T cells are essential for clearance of a lymphocytic choriomeningitis virus (LCMV) infection, but the virus can present a number of unique challenges that antiviral T cells must overcome. Peripheral LCMV infection can lead to rapid cytolytic clearance or chronic viral persistence; central nervous system infection can result in T-cell-dependent fatal meningitis or an asymptomatic carrier state amenable to immunotherapeutic clearance. These diverse outcomes all depend on interactions that require TCR engagement of cognate peptide-MHC complexes. In this review, we explore the diversity in antiviral T-cell behaviors resulting from TCR engagement, beginning with an overview of the immunological synapse and progressing to regulators of TCR signaling that shape the delicate balance between immunopathology and viral clearance.


Loss of BAP1 function leads to EZH2-dependent transformation.
The tumor suppressors BAP1 and ASXL1 interact to form a polycomb deubiquitinase complex that removes monoubiquitin from histone H2A lysine 119 (H2AK119Ub). However, BAP1 and ASXL1 are mutated in distinct cancer types, consistent with independent roles in regulating epigenetic state and malignant transformation. Here we demonstrate that Bap1 loss in mice results in increased trimethylated histone H3 lysine 27 (H3K27me3), elevated enhancer of zeste 2 polycomb repressive complex 2 subunit (Ezh2) expression, and enhanced repression of polycomb repressive complex 2 (PRC2) targets. These findings contrast with the reduction in H3K27me3 levels seen with Asxl1 loss. Conditional deletion of Bap1 and Ezh2 in vivo abrogates the myeloid progenitor expansion induced by Bap1 loss alone. Loss of BAP1 results in a marked decrease in H4K20 monomethylation (H4K20me1). Consistent with a role for H4K20me1 in the transcriptional regulation of EZH2, expression of SETD8-the H4K20me1 methyltransferase-reduces EZH2 expression and abrogates the proliferation of BAP1-mutant cells. Furthermore, mesothelioma cells that lack BAP1 are sensitive to EZH2 pharmacologic inhibition, suggesting a novel therapeutic approach for BAP1-mutant malignancies.


Independent state health surveys: responding to the need for local population health data.
There is high demand for local-level population health data. A national system of state and local data collection would help improve both population health and health care delivery. The primary source of state-level population health data for adults is the Behavioral Risk Factor Surveillance System. However, many states need data on children and adolescents, racial and ethnic subpopulations, consistent estimates for localities, or more in-depth information on key topics than the Behavioral Risk Factor Surveillance System provides. Eleven state health surveys (SHSs) have emerged in an effort to address these gaps. Semistructured telephone interviews were conducted in 2009 with representatives of 9 SHSs. The interviews were recorded, and data were transcribed, organized, and analyzed according to the query structure. This analysis identified (1) the core elements of SHS that have been successful in meeting needs for local data and (2) the processes and strategies used by state officials in creating these surveys. Key findings include the following: (1) SHSs provide concrete data on local health issues that meet the needs of policy makers who wish to adopt evidence-based public health policies; (2) data from SHSs allow researchers to identify issues, apply for grants, and evaluate, assess, and track health indicators; (3) a "champion" is required to build the case for a survey and push through barriers to obtain funding and stakeholder buy-in; and (4) SHSs face challenges such as inconsistent funding and lack of uniform standards. Opportunities to support SHSs include (1) identifying sustained funding sources; (2) providing technical assistance and facilitating training to foster best practices, quality standards, and comparability across states; and (3) supporting an organization for SHS researchers to share resources, information, and experiences.


Left ventricular early myocardial dysfunction after chronic misuse of anabolic androgenic steroids: a Doppler myocardial and strain imaging analysis.
Anabolic androgenic steroids (AAS) are sometimes used by power athletes to improve performance by increasing muscle mass and strength. Recent bioptical data have shown that in athletes under the pharmacological effects of AAS, a focal increase in myocardial collagen content might occur as a repair mechanism against myocardial damage. To investigate the potential underlying left ventricular myocardial dysfunction after chronic misuse of AAS in athletes by use of Doppler myocardial imaging (DMI) and strain rate imaging (SRI). Standard Doppler echocardiography, DMI, SRI and ECG treadmill test were undertaken by 45 bodybuilders, including 20 athletes misusing AAS for at least 5 years (users), by 25 anabolic-free bodybuilders (non-users) and by 25 age-matched healthy sedentary controls, all men. The mean (SD) number of weeks of AAS use per year was 31.3 (6.4) in users, compared with 8.9 (3.8) years in non-users, and the mean weekly dosage of AAS was 525.4 (90.7) mg. The groups were matched for age. Systolic blood pressure was higher in athletes (145 (9) vs 130 (5) mm Hg) than in controls. Left ventricular mass index did not significantly differ between the two groups of athletes. In particular, both users and non-users showed increased wall thickness and relative wall thickness compared with controls, whereas left ventricular ejection fraction, left ventricular end-diastolic diameter and transmitral Doppler indexes were comparable for the three groups. Colour DMI analysis showed significantly lower myocardial early: myocardial atrial diastolic wave ratios in users at the level of the basal interventricular septum (IVS) and left ventricular lateral wall (p<0.01), in comparison with both non-users and controls. In addition, in users, peak systolic left ventricular strain rate and strain were both reduced in the middle IVS (both p<0.001) and in the left ventricular lateral free wall (both p<0.01). By stepwise forward multivariate analyses, the sum of the left ventricular wall thickness (beta coefficient = -0.32, p<0.01), the number of weeks of AAS use per year (beta = -0.42, p<0.001) and the weekly dosage of AAS (beta = -0.48, p<0.001) were the only independent determinants of middle IVS strain rate. In addition, impaired left ventricular strain in users was associated with a reduced performance during physical effort (p<0.001). Several years after chronic misuse of AAS, power athletes show a subclinical impairment of both systolic and diastolic myocardial function, strongly associated with mean dosage and duration of AAS use. The combined use of DMI and SRI may therefore be useful for the early identification of patients with more diffused cardiac involvement, and eventually for investigation of the reversibility of such myocardial effects after discontinuation of the drug.


Ultra-low-power carrier-depletion Mach-Zehnder silicon optical modulator.
We demonstrate a 26 Gbit/s Mach-Zehnder silicon optical modulator. The doping concentration and profile are optimized, and a modulation efficiency with the figure of merit (VπL) of 1.28 V·cm is achieved. We design an 80-nm-wide intrinsic silicon gap between the p-type and n-type doped regions to reduce the capacitance of the diode and prevent the diode from working in a slow diffusion mode. Therefore, the modulator can be driven with a small differential voltage of 0.5 V with no bias. Without the elimination of the dissipated power of the series resistors and the reflected power of the electrical signal, the maximum power consumption is 3.8 mW.


A myriad of miRNA variants in control and Huntington's disease brain regions detected by massively parallel sequencing.
Huntington disease (HD) is a neurodegenerative disorder that predominantly affects neurons of the forebrain. We have applied the Illumina massively parallel sequencing to deeply analyze the small RNA populations of two different forebrain areas, the frontal cortex (FC) and the striatum (ST) of healthy individuals and individuals with HD. More than 80% of the small-RNAs were annotated as microRNAs (miRNAs) in all samples. Deep sequencing revealed length and sequence heterogeneity (IsomiRs) for the vast majority of miRNAs. Around 80-90% of the miRNAs presented modifications in the 3'-terminus mainly in the form of trimming and/or as nucleotide addition variants, while the 5'-terminus of the miRNAs was specially protected from changes. Expression profiling showed strong miRNA and isomiR expression deregulation in HD, most being common to both FC and ST. The analysis of the upstream regulatory regions in co-regulated miRNAs suggests a role for RE1-Silencing Transcription Factor (REST) and P53 in miRNAs downregulation in HD. The putative targets of deregulated miRNAs and seed-region IsomiRs strongly suggest that their altered expression contributes to the aberrant gene expression in HD. Our results show that miRNA variability is a ubiquitous phenomenon in the adult human brain, which may influence gene expression in physiological and pathological conditions.


Long-term prognostic value of 13N-ammonia myocardial perfusion positron emission tomography added value of coronary flow reserve.
The goal of this study was to assess the predictive value of myocardial perfusion imaging with (13)N-ammonia positron emission tomography (PET) and coronary flow reserve (CFR) on long-term prognosis in patients with suspected myocardial ischemia. No prognostic data exist on the predictive value of CFR and (13)N-ammonia PET. Perfusion and CFR were assessed in 256 patients using (13)N-ammonia PET, and follow-up was obtained in 245 (96%) patients. Sixteen early revascularized patients were excluded and 229 were assigned to normal versus abnormal perfusion or normal versus abnormal CFR (<2.0). Major adverse cardiac events (MACE) (cardiac death, nonfatal myocardial infarction, late revascularization, or hospitalization for cardiac reasons) were assessed using the Kaplan-Meier method. Cox proportional hazard regression was used to identify independent predictors for cardiac events. During follow-up (5.4 +/- 2.2 years), 78 patients had at least 1 cardiac event, including 29 cardiac deaths. Abnormal perfusion (n = 126) was associated with a higher incidence of MACE (p < 0.001) and cardiac death (p < 0.05). In patients with normal perfusion, abnormal CFR was independently associated with a higher annual event rate over 3 years compared with normal CFR for MACE (1.4% vs. 6.3%; p < 0.05) and cardiac death (0.5% vs. 3.1%; p < 0.05). In abnormal perfusion, CFR remained predictive throughout the 10-year follow-up (p < 0.001). Perfusion findings in (13)N-ammonia PET and CFR are strong outcome predictors. CFR allows further risk stratification, suggesting a "warranty" period of 3 years if normal CFR is associated with normal perfusion. Conversely, in patients with abnormal perfusion, an impaired CFR has added value for predicting adverse outcomes.


Arm Ergometry to Improve Mobility in Progressive Multiple Sclerosis (AMBOS)-Results of a Pilot Randomized Controlled Trial.
Background: Walking disability is one of the most frequent and burdening symptoms of progressive multiple sclerosis (MS). Most of the exercise intervention studies that showed an improvement in mobility performance were conducted in low to moderately disabled relapsing-remitting MS patients with interventions using the legs. However, MS patients with substantial walking disability hardly can perform these tasks. Earlier work has indicated that aerobic arm training might also improve walking performance and could therefore be a therapeutic option in already moderately disabled progressive MS patients. Methods: Patients with progressive MS and EDSS 4-6.5 were randomized using a computer-generated algorithm list to either a waitlist control group (CG) or an intervention group (IG). The IG performed a 12-week home-based, individualized arm ergometry exercise training program. Maximum walking distance as measured by the 6-min walking test (6MWT) was the primary endpoint. Secondary endpoints included aerobic fitness, other mobility tests, cognitive functioning, as well as fatigue and depression. Results: Of n = 86 screened patients, 53 with moderate disability (mean EDSS 5.5, SD 0.9) were included and data of 39 patients were analyzed. Patients in the IG showed strong adherence to the program with a mean of 67 (SD 26.4) training sessions. Maximum work load (P max) increased in the training group while other fitness indicators did not. Walking distance in the 6MWT improved in both training and waitlist group but not significantly more in trained patients. Similarly, other mobility measures showed no differential group effect. Cognitive functioning remained unchanged. No serious events attributable to the intervention occurred. Conclusion: Although maximum work load improved, 3 months of high-frequency arm ergometry training of low to moderate intensity could not show improved walking ability or cognitive functioning in progressive MS compared to a waitlist CG. The study was registered at www.clinicaltrials.gov (NCT03147105) and funded by the local MS self-help organization.


Remote distribution of a mode-locked pulse train with sub 40-as jitter.
Remote transfer of an ultralow-jitter microwave frequency reference signal is demonstrated using the pulse trains generated by a mode-locked fiber laser. The timing jitter in a ~ 30-m fiber link is reduced to 38 attoseconds (as) integrated over a bandwidth from 1 Hz to 10 MHz via active stabilization which represents a significant improvement over previously reported jitter performance. Our approach uses an all-optical generation of the synchronization error signal and an accompanying out-of-loop optical detection technique to verify the jitter performance.


Prairie dogs avoid extreme inbreeding.
Black-tailed prairie dogs (Rodentia: Sciuridae: Cynomys ludovicianus) live in colonies composed of contiguous but separate family groups called coteries. During the 6 years that individuals in a colony were observed, they almost nevermated with close genetic relatives. Inbreeding is avoided in four ways: (i) a young male usually leaves his natal coterie before breeding, but his female relatives remain; (ii) an adult male usually leaves his breeding coterie before his daughters mature; (iii) a young female is less likely to come into estrus if her father is in her coterie; and (iv) an estrous female behaviorally avoids mating with a father, son, or brother in her coterie.


Polypropylene/Layered Double Hydroxide (LDH) Nanocomposites: Influence of LDH Particle Size on the Crystallization Behavior of Polypropylene.
Highly dispersed isotactic polypropylene (iPP) nanocomposites were prepared by incorporating two different sized Mg-Al LDH nanoparticles with different loadings from 1 to 10 wt % using a modified solvent mixing method. Larger sized LDH nanoparticles (∼3-4 μm) were prepared from the gel form of Mg-Al LDH, and the smaller sized nanoparticles (∼50-200 nm) were prepared by sonication of as-synthesized LDH particles. Such obtained LDH nanoparticles were carefully characterized using wide-angle X-ray diffraction (WAXD), transmission electron microscopy, and scanning electron microscopy. WAXD and atomic force microscopy results indicate that the LDH nanoparticles were highly dispersed in the iPP matrix. The influence of LDH nanoparticles size and concentration on the thermal stability, spherulitic morphology, melting behavior, isothermal crystallization kinetics, and lamellar structure of iPP were investigated. Incorporation of low loadings of sonicated LDH particles (e.g., 1-2.5 wt %) show substantial effect on thermal stability, spherulite size, crystallinity, and crystallization half-time and lamellar morphology of iPP compared to the pure iPP and that of nanocomposites with larger LDH particles with same loadings. The better nucleation ability of iPP in the presence of sonicated LDH can be attributed to the high surface area of LDH nanoparticles along with its better dispersibility within the polymer matrix. The incorporation of LDH nanoparticles does not change the crystallization growth mechanism and crystal structure of iPP.


Psychedelic philanthropy: The nonprofit sector and Timothy Leary's 1960s psychedelic movement.
Little has been written on the financial support behind Timothy Leary's unorthodox research into mind-altering drugs like LSD and psilocybin and his subsequent psychedelic movement. Indeed several individuals and organizations helped the psychedelic cause by directly funding Leary's ventures, offering legal and logistic assistance, and organizing fund-raising campaigns. I argue that classic philanthropic attitudes and wealthy patrons played a major supporting role for Leary's psychedelic movement in the first part of the decade and that the changes in Leary's research objectives and his transition from academic to LSD guru were accompanied by changes in the patterns of support that occurred throughout the 1960s. This paper also connects Leary's legal troubles in the second part of the decade with the rise of the movement to legalize cannabis and points to historical continuity by looking at contemporary endeavors to fund psychedelic research.


A composition-dependent molecular clutch between T cell signaling condensates and actin.
During T cell activation, biomolecular condensates form at the immunological synapse (IS) through multivalency-driven phase separation of LAT, Grb2, Sos1, SLP-76, Nck, and WASP. These condensates move radially at the IS, traversing successive radially-oriented and concentric actin networks. To understand this movement, we biochemically reconstituted LAT condensates with actomyosin filaments. We found that basic regions of Nck and N-WASP/WASP promote association and co-movement of LAT condensates with actin, indicating conversion of weak individual affinities to high collective affinity upon phase separation. Condensates lacking these components were propelled differently, without strong actin adhesion. In cells, LAT condensates lost Nck as radial actin transitioned to the concentric network, and engineered condensates constitutively binding actin moved aberrantly. Our data show that Nck and WASP form a clutch between LAT condensates and actin in vitro and suggest that compositional changes may enable condensate movement by distinct actin networks in different regions of the IS.


Endemic Cryptosporidium and Giardia lamblia infections in a Thai orphanage.
We conducted a point prevalence survey for enteric protozoa in 205 institutionalized orphans 1-61 months of age in Bangkok, Thailand. Cryptosporidium was identified in 17 children (8%), Giardia lamblia in 42 (20%), and 3 children (1%) had both parasites. At the time of diagnosis, diarrheal symptoms were present in a minority of subjects: 36% of children with Cryptosporidium alone, 10% with G. lamblia alone, and in 20% of those with neither parasite. Although chronic nutritional status (height/age) was similar in all groups, acute nutritional status (weight/height) was lower only in children with Cryptosporidium (Z score = -1.39 +/- 0.13) compared with children with G. lamblia (mean Z score +/- SEM = -0.56 +/- 0.26) or neither parasite (Z score = -0.78 +/- 0.13; P = 0.05). Detectable levels of Cryptosporidium-specific IgG antibodies by ELISA were identified in 15 of 16 Thai children with Cryptosporidium and in 17 of 19 Thai children without Cryptosporidium (mean OD +/- SEM = 1.27 +/- 0.18 vs. 1.06 +/- 0.13, respectively), but in only 1 of 18 sera from toddlers in day-care centers in Denver, CO (OD = 0.128 +/- 0.03). Although neither infection with Cryptosporidium nor G. lamblia was consistently associated with acute diarrheal symptoms, Cryptosporidium was more often associated with depressed acute nutritional status than G. lamblia. The high prevalence of specific antibodies to Cryptosporidium in Thai orphans suggests an association between high rates of exposure with asymptomatic excretion of the parasites.


Vitamin D physiology.
Vitamin D3 is synthesized in the skin during summer under the influence of ultraviolet light of the sun, or it is obtained from food, especially fatty fish. After hydroxylation in the liver into 25-hydroxyvitamin D (25(OH)D) and kidney into 1,25-dihydroxyvitamin D (1,25(OH)2D), the active metabolite can enter the cell, bind to the vitamin D-receptor and subsequently to a responsive gene such as that of calcium binding protein. After transcription and translation the protein is formed, e.g. osteocalcin or calcium binding protein. The calcium binding protein mediates calcium absorption from the gut. The production of 1,25(OH)2D is stimulated by parathyroid hormone (PTH) and decreased by calcium. Risk factors for vitamin D deficiency are premature birth, skin pigmentation, low sunshine exposure, obesity, malabsorption and advanced age. Risk groups are immigrants and the elderly. Vitamin D status is dependent upon sunshine exposure but within Europe, serum 25(OH)D levels are higher in Northern than in Southern European countries. Severe vitamin D deficiency causes rickets or osteomalacia, where the new bone, the osteoid, is not mineralized. Less severe vitamin D deficiency causes an increase of serum PTH leading to bone resorption, osteoporosis and fractures. A negative relationship exists between serum 25(OH)D and serum PTH. The threshold of serum 25(OH)D, where serum PTH starts to rise is about 75nmol/l according to most surveys. Vitamin D supplementation to vitamin D-deficient elderly suppresses serum PTH, increases bone mineral density and may decrease fracture incidence especially in nursing home residents. The effects of 1,25(OH)2D and the vitamin D receptor have been investigated in patients with genetic defects of vitamin D metabolism and in knock-out mouse models. These experiments have demonstrated that for active calcium absorption, longitudinal bone growth and the activity of osteoblasts and osteoclasts both 1,25(OH)2D and the vitamin D receptor are essential. On the other side, bone mineralization can occur by high ambient calcium concentration, so by high doses of oral calcium or calcium infusion. The active metabolite 1,25(OH)2D has its effects through the vitamin D receptor leading to gene expression, e.g. the calcium binding protein or osteocalcin or through a plasma membrane receptor and second messengers such as cyclic AMP. The latter responses are very rapid and include the effects on the pancreas, vascular smooth muscle and monocytes. Muscle cells contain vitamin D receptor and several studies have demonstrated that serum 25(OH)D is related to physical performance. The active metabolite 1,25(OH)2D has an antiproliferative effect and downregulates inflammatory markers. Extrarenal synthesis of 1,25(OH)2D occurs under the influence of cytokines and is important for the paracrine regulation of cell differentiation and function. This may explain that vitamin D deficiency can play a role in the pathogenesis of auto-immune diseases such as multiple sclerosis and diabetes type 1, and cancer. In conclusion, the active metabolite 1,25(OH)2D has pleiotropic effects through the vitamin D receptor and vitamin D responsive elements of many genes and on the other side rapid non-genomic effects through a membrane receptor and second messengers. Active calcium absorption from the gut depends on adequate formation of 1,25(OH)2D and an intact vitamin D receptor. Bone mineralization mainly depends on ambient calcium concentration. Vitamin D metabolites may play a role in the prevention of auto-immune disease and cancer.


Female undergraduate's perceptions of intrusive behavior in 12 countries.
The present study examines young women's (N = 1,734) perceptions of the unacceptability of 47 intrusive activities enacted by men. Female undergraduate psychology students from 12 countries (Armenia, Australia, England, Egypt, Finland, India, Indonesia, Italy, Japan, Portugal, Scotland, Trinidad) indicated which of 47 intrusive activities they considered to be unacceptable. Responses were compared with parasite-stress values, a measure of global gender equality and Hofstede's dimensions of national cultures. There was no unanimous agreement on any of the items, even for those relating to forced sexual violence. Cluster analysis yielded four clusters: "Aggression and surveillance" (most agreement that the constituent items were unacceptable), "Unwanted attention," "Persistent courtship and impositions," and "Courtship and information seeking" (least agreement that the constituent items were unacceptable). There were no significant relationships between the "Aggression and surveillance" or "Courtship and information seeking" clusters and the measure of gender equality, Hofstede's dimensions of national cultures or the measure of parasite stress. For the "Unwanted attention" and "Persistent courtship and impositions" clusters, women residing in countries with higher gender inequality and higher parasite-stress were less accepting of behavior associated with uncommitted sexual relations, and women in more individualistic societies with higher levels of gender equality were less accepting of monitoring activities. Culture may take precedence over personal interpretations of the unacceptability of intrusive behavior that is not obviously harmful or benign in nature.


Zoledronate prevents lactation induced bone loss and results in additional post-lactation bone mass in mice.
In rodents, lactation is associated with a considerable and very rapid bone loss, which almost completely recovers after weaning. The aim of the present study was to investigate whether the bisphosphonate Zoledronate (Zln) can inhibit lactation induced bone loss, and if Zln interferes with recovery of bone mass after lactation has ceased. Seventy-six 10-weeks-old NMRI mice were divided into the following groups: Baseline, Pregnant, Lactation, Lactation+Zln, Recovery, Recovery+Zln, and Virgin Control (age-matched). The lactation period was 12days, then the pups were removed, and thereafter recovery took place for 28days. Zln, 100μg/kg, was given s.c. on the day of delivery, and again 4 and 8days later. Mechanical testing, μCT, and dynamic histomorphometry were performed. At L4, lactation resulted in a substantial loss of bone strength (-55% vs. Pregnant, p<0.01), BV/TV (-40% vs. Pregnant, p<0.01), and trabecular thickness (Tb.Th) (-29% vs. Pregnant, p<0.001). Treatment with Zln completely prevented lactation induced loss of bone strength, BV/TV, and Tb.Th at L4. Full recovery of micro-architectural and mechanical properties was found 28days after weaning in vehicle-treated mice. Interestingly, the recovery group treated with Zln during the lactation period had higher BV/TV (+45%, p<0.01) and Tb.Th (+16%, p<0.05) compared with virgin controls. Similar results were found at the proximal tibia and femur. This indicates that Zln did not interfere with the bone formation taking place after weaning. On this background, we conclude that post-lactation bone formation is not dependent on a preceding lactation induced bone loss.


Energy Landscapes Reveal Agonist Control of G Protein-Coupled Receptor Activation via Microswitches.
Agonist binding to G protein-coupled receptors (GPCRs) leads to conformational changes in the transmembrane region that activate cytosolic signaling pathways. Although high-resolution structures of different receptor states are available, atomistic details of allosteric signaling across the membrane remain elusive. We calculated free energy landscapes of β2 adrenergic receptor activation using atomistic molecular dynamics simulations in an optimized string of swarms framework, which shed new light on how microswitches govern the equilibrium between conformational states. Contraction of the extracellular binding site in the presence of the agonist BI-167107 is obligatorily coupled to conformational changes in a connector motif located in the core of the transmembrane region. The connector is probabilistically coupled to the conformation of the intracellular region. An active connector promotes desolvation of a buried cavity, a twist of the conserved NPxxY motif, and an interaction between two conserved tyrosines in transmembrane helices 5 and 7 (Y-Y motif), which lead to a larger population of active-like states at the G protein binding site. This coupling is augmented by protonation of the strongly conserved Asp792.50. The agonist binding site hence communicates with the intracellular region via a cascade of locally connected microswitches. Characterization of these can be used to understand how ligands stabilize distinct receptor states and contribute to development drugs with specific signaling properties. The developed simulation protocol can likely be transferred to other class A GPCRs.


Real time bleaching of methylene blue or thionine sensitized gelatin.
The kinetics of bleaching of methylene blue or thionine sensitized gelatin plates are investigated with a singlebeam exposure experiment. It is found that thionine bleaching can be modeled by a first-order theory; whereas, a higher-order theory is found necessary to explain methylene blue bleaching.


Trying to Do What Is Best: A Qualitative Study of Maternal-Infant Bonding and Neonatal Abstinence Syndrome.
The maternal experience of caring for and bonding with infants affected by neonatal abstinence syndrome (NAS) has not been adequately characterized. This study was designed to describe mothers' experiences of, supports for, and barriers to bonding with infants with NAS. Semistructured interviews were coded using computer-assisted thematic content analysis. A code co-occurrence model was used to visualize the relationships between themes. Thirteen mothers of infants with NAS participated. Trying to Do What Is Best emerged as the overarching theme with which several subthemes co-occurred. Subthemes that captured mothers loving their infants and bonding, feeling supported by the infants' fathers, feeling supported in the community, and receiving information from hospital staff were associated with mothers' trying to do what is best. Barriers to trying to do what is best included feeling unsupported in the community, guilt about taking medications or substances during pregnancy, feeling judged, and infant withdrawal. Specific implications for practice may be derived from the mothers' criticisms of NAS assessment tools. Mothers highlighted the value of reassurance and education from providers and the uniquely nonjudgmental support received from peers and male coparents. There is a lack of information about maternal-infant bonding in dyads affected by NAS and factors that contribute to parental loss of custody. Qualitative, quantitative, and mixed-methods studies in diverse populations might help researchers better understand the long-term outcomes of NAS and develop interventions that decrease family separation.


Insulin-mediated signaling promotes proliferation and survival of glioblastoma through Akt activation.
Metabolic complications such as obesity, hyperglycemia, and type 2 diabetes are associated with poor outcomes in patients with glioblastoma. To control peritumoral edema, use of chronic high-dose steroids in glioblastoma patients is common, which can result in de novo diabetic symptoms. These metabolic complications may affect tumors via profound mechanisms, including activation of insulin receptor (InsR) and the related insulin-like growth factor 1 receptor (IGF1R) in malignant cells. In the present study, we assessed expression of InsR in glioblastoma surgical specimens and glioblastoma response to insulin at physiologically relevant concentrations. We further determined whether genetic or pharmacological targeting of InsR affected oncogenic functions of glioblastoma in vitro and in vivo. We showed that InsR was commonly expressed in glioblastoma surgical specimens and xenograft tumor lines, with mitogenic isoform-A predominating. Insulin at physiologically relevant concentrations promoted glioblastoma cell growth and survival, potentially via Akt activation. Depletion of InsR impaired cellular functions and repressed orthotopic tumor growth. The absence of InsR compromised downstream Akt activity, but yet stimulated IGF1R expression. Targeting both InsR and IGF1R with dual kinase inhibitors resulted in effective blockade of downstream signaling, loss of cell viability, and repression of xenograft tumor growth. Taken together, our work suggests that glioblastoma is sensitive to the mitogenic functions of insulin, thus significant insulin exposure imposes risks to glioblastoma patients. Additionally, dual inhibition of InsR and IGF1R exhibits promise for treating glioblastoma.


Investigating the impact of observation errors on the statistical performance of network-based diffusion analysis.
Experiments in captivity have provided evidence for social learning, but it remains challenging to demonstrate social learning in the wild. Recently, we developed network-based diffusion analysis (NBDA; 2009) as a new approach to inferring social learning from observational data. NBDA fits alternative models of asocial and social learning to the diffusion of a behavior through time, where the potential for social learning is related to a social network. Here, we investigate the performance of NBDA in relation to variation in group size, network heterogeneity, observer sampling errors, and duration of trait diffusion. We find that observation errors, when severe enough, can lead to increased Type I error rates in detecting social learning. However, elevated Type I error rates can be prevented by coding the observed times of trait acquisition into larger time units. Collectively, our results provide further guidance to applying NBDA and demonstrate that the method is more robust to sampling error than initially expected. Supplemental materials for this article may be downloaded from http://lb.psychonomic-journals.org/content/supplemental.


Expression of the CD34 gene in vascular endothelial cells.
All seven of a set of CD34 monoclonal antibodies that recognize epitopes on an approximately 110 Kd glycoprotein on human hemopoietic progenitor cells also bind to vascular endothelium. Capillaries of most tissues are CD34 positive, as are umbilical artery and, to a lesser extent, vein, but the endothelium of most large vessels and the endothelium of placental sinuses are not. Angioblastoma cells and parafollicular mesenchymal cells in fetal skin are also CD34 positive, as are some stromal elements. An approximately 110 Kd protein can be identified by Western blot analysis with CD34 antibodies in detergent extracts of freshly isolated umbilical vessel endothelial cells, and CD34 mRNA is present in cultured umbilical vein cells as well as other tissues rich in vascular endothelium (breast, placenta). These data indicate that the binding of CD34 antibodies to vascular endothelium is to the CD34 gene product, and not to crossreactive epitopes. Despite the presence of CD34 mRNA in cultured, proliferating endothelial cells, the latter do not bind CD34 antibodies. In addition, CD34 antigen cannot be upregulated by growth factors. We conclude that under these conditions, CD34 protein is downregulated or processed into another form that is unreactive with CD34 antibodies. Electron microscopy of umbilical artery, breast, and kidney capillary vessels reveals that in all three sites, CD34 molecules are concentrated on membrane processes, many of which interdigitate between adjacent endothelial cells. However, well-established endothelial cell contacts with tight junctions are CD34 negative. CD34 may function as an adhesion molecule on both endothelial cells and hematopoietic progenitors.


Learning to perform role-filler binding with schematic knowledge.
Through specific experiences, humans learn the relationships that underlie the structure of events in the world. Schema theory suggests that we organize this information in mental frameworks called "schemata," which represent our knowledge of the structure of the world. Generalizing knowledge of structural relationships to new situations requires role-filler binding, the ability to associate specific "fillers" with abstract "roles." For instance, when we hear the sentence Alice ordered a tea from Bob, the role-filler bindings customer:Alice, drink:tea and barista:Bob allow us to understand and make inferences about the sentence. We can perform these bindings for arbitrary fillers-we understand this sentence even if we have never heard the names Alice, tea, or Bob before. In this work, we define a model as capable of performing role-filler binding if it can recall arbitrary fillers corresponding to a specified role, even when these pairings violate correlations seen during training. Previous work found that models can learn this ability when explicitly told what the roles and fillers are, or when given fillers seen during training. We show that networks with external memory learn to bind roles to arbitrary fillers, without explicitly labeled role-filler pairs. We further show that they can perform these bindings on role-filler pairs that violate correlations seen during training, while retaining knowledge of training correlations. We apply analyses inspired by neural decoding to interpret what the networks have learned.


Association between perceived insufficient sleep, frequent mental distress, obesity and chronic diseases among US adults, 2009 behavioral risk factor surveillance system.
Although evidence suggests that poor sleep is associated with chronic disease, little research has been conducted to assess the relationships between insufficient sleep, frequent mental distress (FMD ≥14 days during the past 30 days), obesity, and chronic disease including diabetes mellitus, coronary heart disease, stroke, high blood pressure, asthma, and arthritis. Data from 375,653 US adults aged ≥ 18 years in the 2009 Behavioral Risk Factor Surveillance System were used to assess the relationships between insufficient sleep and chronic disease. The relationships were further examined using a multivariate logistic regression model after controlling for age, sex, race/ethnicity, education, and potential mediators (FMD and obesity). The overall prevalence of insufficient sleep during the past 30 days was 10.4% for all 30 days, 17.0% for 14-29 days, 42.0% for 1-13 days, and 30.6% for zero day. The positive relationships between insufficient sleep and each of the six chronic disease were significant (p < 0.0001) after adjustment for covariates and were modestly attenuated but not fully explained by FMD. The relationships between insufficient sleep and both diabetes and high blood pressure were also modestly attenuated but not fully explained by obesity. Assessment of sleep quantity and quality and additional efforts to encourage optimal sleep and sleep health should be considered in routine medical examinations. Ongoing research designed to test treatments for obesity, mental distress, or various chronic diseases should also consider assessing the impact of these treatments on sleep health.


Increased alpha-synuclein tear fluid levels in patients with Parkinson's disease.
The objective of the study was to estimate if altered levels of alpha-synuclein can be detected in tear fluid of patients with Parkinson's disease (PD). Therefore, tear fluid samples of 75 PD patients, 75 control subjects and 31 atypical Parkinsonian patients were collected and analyzed in triplicates using an ultra-sensitive single molecule array (SIMOA) system and applying a human alpha-synuclein immunoassay. In PD, levels of total soluble alpha-synuclein were significantly increased compared to control subjects (p = 0.03; AUC PD vs. controls 0.60). There was no difference comparing PD patients stratified by Hoehn & Yahr stages and atypical Parkinsonian syndromes stratified by tauopathies and non-PD-synucleinopathies against each other (p > 0.05). In conclusion, alpha-synuclein can be detected and quantified in tear fluid, revealing small but significant differences in total alpha-synuclein levels between PD and control subjects. Tear fluid can be collected non-invasively and risk-free, therefore presenting a promising source for further biomarker research.


The Associations of Breast Feeding with Infant Growth and Body Mass Index to 16 years: 'Children of 1997'.
Controversial findings concerning associations of breast feeding with growth have been reported. This study examined whether the associations of breast feeding with early growth trajectories and body mass index to 16 years differed by sex or age. In Hong Kong's 'Children of 1997' population-representative birth cohort, contemporaneously reported breast-feeding status in the first 3 months was classified as exclusive breast feeding (BF) (n = 470), mixed feeding (MF) (n = 2693), and formula feeding (FF) (n = 4204). Adjusted sex- and age-specific associations of breast feeding with infant growth (gains in weight-for-age z scores (WAZ), length/height-for-age z scores (LAZ), and body-mass-index-for-age z score (BAZ) based on the World Health Organization standards/references from birth to 36 months) were assessed using linear regression and mixed modelling, respectively. Adjusted sex-specific associations of breast feeding with average BAZ from 3 months to 16 years were assessed using generalized estimating equation. Potential confounders were maternal and infant characteristics, and household income. Among 7367 children, associations of breast feeding with infant growth did not vary by sex, but WAZ gains varied by age. Greater WAZ gains were observed in BF than FF infants from 0 to 3 months but in FF than BF infants from 3 to 9 months. Breast feeding was not associated with overall BAZ from 3 months to 16 years, with no differences by sex. Our findings suggest that breast feeding may only have short-term effects on growth. Further studies of the role of breast feeding in other metabolic diseases may be needed.


Immunophenotypic and genotypic characteristics of chronic myelogenous leukemia in blast crisis.
Chronic myelogenous leukemia (CML) may transform into blast crisis (BC) if not properly treated. Among patients with transformation, 20% to 30% will develop BC with lymphoid-associated antigens (Ly-BC), and the remaining cases with myeloid-associated antigens (My-BC) or with both (Mix-BC). In this study, we investigated the lineage of blast cells in CML-BC using immunophenotypic and genetic analyses and analyzed the prognostic significance of genotypic change in CML-BC. Twenty-one patients with CML-BC diagnosed at the Taipei Veterans General Hospital from 1982 to 1992 were included. Immunophenotyping was done by using the avidin-biotin immunoperoxidase technique. Genetic analyses were carried out by using Southern Blot hybridization. The prognostic influence of genotypic change was analyzed. Thirteen patients (61.9%) expressed myeloid-associated antigens, one patient (4.8%) expressed megakaryoblast-associated antigens, four patients (19%) expressed B lymphoid-associated antigens and three patients (14.3%) expressed both myeloid and B lymphoid antigens. Clonal rearrangement of the immunoglobulin heavy chain (IgH) gene was found in six cases. Among them, four expressed B lymphoid markers only and two expressed both myeloid and B lymphoid markers. Patients with clonal IgH gene rearrangement tended to have a better response to chemotherapy (50% vs 8.3%, p = 0.08) and significantly longer survival (median survival, 5 months vs 3 months, p < 0.05) than did those with a germline configuration. Clonal rearrangement of the IgH gene was found mostly in cases of Ly-BC and Mix-BC. We found that CML-BC with clonal rearrangement of the IgH gene had a more favorable prognosis than in cases with a germline configuration.


A Single Simulated Heliox Dive Modifies Endothelial Function in the Vascular Wall of ApoE Knockout Male Rats More Than Females.
The number of divers is rising every year, including an increasing number of aging persons with impaired endothelial function and concomitant atherosclerosis. While diving is an independent modulator of endothelial function, little is known about how diving affects already impaired endothelium. In this study, we questioned whether diving exposure leads to further damage of an already impaired endothelium. A total of 5 male and 5 female ApoE knockout (KO) rats were exposed to simulated diving to an absolute pressure of 600 kPa in heliox gas (80% helium, 20% oxygen) for 1 h in a dry pressure chamber. 10 ApoE KO rats (5 males, 5 females) and 8 male Sprague-Dawley rats served as controls. Endothelial function was examined in vitro by isometric myography of pulmonary and mesenteric arteries. Lipid peroxidation in blood plasma, heart and lung tissue was used as measures of oxidative stress. Expression and phosphorylation of endothelial NO synthase were quantified by Western blot. A single simulated dive was found to induce endothelial dysfunction in the pulmonary arteries of ApoE KO rats, and this was more profound in male than female rats. Endothelial dysfunction in males was associated with changing in production or bioavailability of NO; while in female pulmonary arteries an imbalance in prostanoid signaling was observed. No effect of diving was found on mesenteric arteries from rats of either sex. Our findings suggest that changes in endothelial dysfunction were specific for pulmonary circulation. In future, human translation of these findings may suggest caution for divers who are elderly or have prior reduced endothelial function.


Structural determinants of slippage-mediated mutations by human immunodeficiency virus type 1 reverse transcriptase.
Single-base deletions at nucleotide runs or -1 frameshifting by human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) result from template slippage during polymerization. In crystal structures of HIV-1 RT complexed with DNA-DNA template-primer, the palm subdomain in the template cleft contacts the template backbone near the proposed site of slippage via the Glu(89) side chain. We investigated the role of Glu(89) in frameshifting by perturbing this interaction. Substitutions with Asp, Gly, Ala, Val, Ser, Thr, Asn, or Lys were created in recombinant HIV RT, and frameshift frequencies of the resulting mutant RTs were measured. All substitutions led to reduced -1 frameshifting by HIV-1 RT (2-40-fold). Interestingly, the suppression of -1 frameshifting frequently coincided with an enhancement of +1 frameshifting (3-47-fold) suggesting that Glu(89) can influence the slippage of both strands. Glu(89) substitutions also led to reduced rates of dNTP misincorporation that paralleled reductions in -1 frameshifting, suggesting a common structural mechanism for both classes of RT error. Our results reveal a major influence of Glu(89) on slippage-mediated errors and dNTP incorporation fidelity. The crystal structure of HIV-1 RT reveals a salt bridge between Glu(89) and Lys(154), which may facilitate -1 frameshifting; this concept is supported by the observed reduction in -1 frameshifting for K154A and K154R mutants.


South Pacific Islanders resist type 2 diabetes: comparison of aerobic and resistance training.
The purpose of this study was to evaluate the effectiveness of two exercise modalities for improving glycosylated hemoglobin (HbA1c) and associated clinical outcomes in Polynesian adults diagnosed with type 2 diabetes and visceral obesity. Twenty-six adults were randomized to receive resistance training or aerobic training, 3×/week, for 16 weeks. Dependent variables collected before and after intervention included: diabetes markers including HbA1c, blood lipids, relevant cytokines (C-reactive protein, adiponectin), and anthropometric and hemodynamic indices. Eighteen participants (72% female; age: 49.3 ± 5.3 years; waist circumference: 128.7 ± 18.7 cm) completed the intervention and follow-up assessments. Body mass index in the whole cohort at baseline indicated Class III (morbid) obesity (43.8 ± 9.5 kg/m(2)). Compliance to training was 73 ± 19 and 67 ± 18% in the aerobic and resistance training groups, respectively. HbA1c remained elevated in both groups after training. Aerobic training reduced systolic and diastolic blood pressure and increased serum triglycerides (all P < 0.05). No other exercise-induced adaptations were noted within or between groups. Post hoc analysis using pooled data indicated that higher adherence to training (≥75% attendance, n = 8) significantly reduced waist circumference (P < 0.001) and tended to reduce body weight and fasting insulin (all P ≤ 0.11) versus lower adherence (<75% attendance, n = 10). In conclusion, this study did not demonstrate an improvement in HbA1c with exercise in morbidly obese Polynesian people. Future investigations involving exercise regimens that are more practicable and which involve greater frequency and duration of training may be required to induce significant and clinically meaningful adaptations in this unique diabetes population.


A surface-based analysis of language lateralization and cortical asymmetry.
Among brain functions, language is one of the most lateralized. Cortical language areas are also some of the most asymmetrical in the brain. An open question is whether the asymmetry in function is linked to the asymmetry in anatomy. To address this question, we measured anatomical asymmetry in 34 participants shown with fMRI to have language dominance of the left hemisphere (LLD) and 21 participants shown to have atypical right hemisphere dominance (RLD). All participants were healthy and left-handed, and most (80%) were female. Gray matter (GM) volume asymmetry was measured using an automated surface-based technique in both ROIs and exploratory analyses. In the ROI analysis, a significant difference between LLD and RLD was found in the insula. No differences were found in planum temporale (PT), pars opercularis (POp), pars triangularis (PTr), or Heschl's gyrus (HG). The PT, POp, insula, and HG were all significantly left lateralized in both LLD and RLD participants. Both the positive and negative ROI findings replicate a previous study using manually labeled ROIs in a different cohort [Keller, S. S., Roberts, N., Garcia-Finana, M., Mohammadi, S., Ringelstein, E. B., Knecht, S., et al. Can the language-dominant hemisphere be predicted by brain anatomy? Journal of Cognitive Neuroscience, 23, 2013-2029, 2011]. The exploratory analysis was accomplished using a new surface-based registration that aligns cortical folding patterns across both subject and hemisphere. A small but significant cluster was found in the superior temporal gyrus that overlapped with the PT. A cluster was also found in the ventral occipitotemporal cortex corresponding to the visual word recognition area. The surface-based analysis also makes it possible to disentangle the effects of GM volume, thickness, and surface area while removing the effects of curvature. For both the ROI and exploratory analyses, the difference between LLD and RLD volume laterality was most strongly driven by differences in surface area and not cortical thickness. Overall, there were surprisingly few differences in GM volume asymmetry between LLD and RLD indicating that gross morphometric asymmetry is only subtly related to functional language laterality.


Evaluation of internal consistency of the epidermolysis bullosa oropharyngeal severity score (EBOS).
To evaluate the internal consistency of the epidermolysis bullosa oropharyngeal severity score (EBOS). Data from 92 patients of varying EB types/sub-types already described in a previous multi-center study were re-analyzed via the coefficient Cronbach's α (CR-α). Additionally, the corrected item total correlation between each item and the items' overall score with Pearson's product-moment correlation (ρ) was calculated. The alpha coefficient for the mean total score of 17 items is 0.941. The inter-observer reliability for disease severity score was excellent for oral medicine specialist (α = 0.924) and dermatologist (α = 0.916) and the intra-observer reliability was good at Time 1 (α = 0.895) and Time 2 (α = 0.897). The analysis of CR-α per single item revealed that alpha was greater than 0.904 for disease activity and 0.743 for structural damage, after the elimination of four items for oral medicine specialist and greater than 0.898 for disease activity and 0.769 for structural damage after the elimination of five items for dermatologist. Similarly the analysis of the corrected items-EBOS correlation showed that the same items do not correlate very well (ρ < 0.4) with the overall EBOS. The EBOS turned out to have a strong and reliable internal consistency, as the majority of the EBOS' items were consistent with each other.


Targeting interventions for ethnic minority and low-income populations.
Although rates of childhood obesity among the general population are alarmingly high, they are higher still in ethnic minority and low-income communities. The disparities pose a major challenge for policymakers and practitioners planning strategies for obesity prevention. In this article Shiriki Kumanyika and Sonya Grier summarize differences in childhood obesity prevalence by race and ethnicity and by socioeconomic status. They show how various environmental factors can have larger effects on disadvantaged and minority children than on their advantaged white peers-and thus contribute to disparities in obesity rates. The authors show, for example, that low-income and minority children watch more television than white, non-poor children and are potentially exposed to more commercials advertising high-calorie, low-nutrient food during an average hour of TV programming. They note that neighborhoods where low-income and minority children live typically have more fast-food restaurants and fewer vendors of healthful foods than do wealthier or predominantly white neighborhoods. They cite such obstacles to physical activity as unsafe streets, dilapidated parks, and lack of facilities. In the schools that low-income and minority children attend, however, they see opportunities to lead the way to effective obesity prevention. Finally, the authors examine several aspects of the home environment-breast-feeding, television viewing, and parental behaviors-that may contribute to childhood obesity but be amenable to change through targeted intervention. Kumanyika and Grier point out that policymakers aiming to prevent obesity can use many existing policy levers to reach ethnic minority and low-income children and families: Medicaid, the State Child Health Insurance Program, and federal nutrition "safety net" programs. Ultimately, winning the fight against childhood obesity in minority and low-income communities will depend on the nation's will to change the social and physical environments in which these communities exist.


Magnetic Resonance Enterography Findings in Crohn's disease in the Pediatric Population and Correlation with Fluoroscopic and Multidetector Computed Tomographic Techniques.
Traditionally, fluoroscopic examinations such as enteroclysis, upper GI studies, and small bowel follow through exams have been the procedures of choice in evaluating inflammatory bowel disease (IBD) in pediatric populations. With the advent of multidetector computed tomography (MDCT), it has subsequently become a complementary examination in imaging inflammatory bowel disease. A major advantage of MDCT over fluoroscopic examination is its ability to directly visualize bowel mucosa, as well as demonstrate extra-enteric complications of IBD such as abscesses, fistulae, and sinus tracts. The major disadvantage of CT however is exposure to ionizing radiation, especially in IBD patients of the pediatric age group who maybe repeatedly imaged due to exacerbations. As a result, magnetic resonance enterography (MRE) is becoming increasingly important in the evaluation and follow-up of pediatric patients with IBD. This pictorial essay will summarize the multi-modality imaging findings of IBD with emphasis on MRE including the imaging protocol and procedure. For the purposes of this article, patients less than 17 years of age have been considered to represent the pediatric population.


Expression of a synthetic gene encoding canine milk lysozyme in Escherichia coli and characterization of the expressed protein.
A high-expression plasmid of the canine milk lysozyme, which belongs to the family of calcium-binding lysozymes, was constructed in order to study its physico-chemical properties. Because the cDNA sequence of the protein has not yet been determined, a 400 base-pair gene encoding canine milk lysozyme was first designed on the basis of the known amino acid sequence. The gene was constructed by an enzymatic assembly of 21 chemically synthesized oligonucleotides and inserted into an Escherichia coli expression vector by stepwise ligation. The expression plasmid thus constructed was transformed into BL21(DE3)/pLysS cells. The gene product accumulated as inclusion bodies in an insoluble fraction. Recombinant canine milk lysozyme was obtained by purification and refolding of the product and showed the same characteristics in terms of bacteriolytic activity and far- and near-UV circular dichroism spectra as the authentic protein. The NMR spectra of refolded lysozyme were also characteristic of a native globular protein. It was concluded that recombinant canine milk lysozyme was folded into the correct native structure. Moreover, the thermal unfolding profiles of the refolded recombinant lysozyme showed a stable equilibrium intermediate, indicating that the molten globule state of this protein was extraordinarily stable. This expression system of canine milk lysozyme will enable biophysical and structural studies of this protein to be extended.


Template-assisted nanostructure fabrication by glancing angle deposition: a molecular dynamics study.
In the present work, we investigate the pre-existing template-assisted glancing angle deposition of Al columnar structures on Cu substrate by means of molecular dynamics simulations, with a focus on examining the effect of deposition-induced template deformation on the morphologies of the fabricated structures. Our simulations demonstrate that the pre-existing templates significantly intensify the shadowing effect, which thus facilitates the formation of columnar structures under small deposition flux. The underlying deformation modes of the templates under different deposition configurations are analyzed and are correlated to the geometrical characteristics of the columnar structures. It is found that the template height-dependent deformation behavior of the templates strongly influences the morphologies of the fabricated columnar structures. Our findings provide design and fabrication guidelines for the fabrication of one-dimensional nanostructures by the template-assisted deposition technique.


ROS open roads to roundworm infection.
The rapid production of reactive oxygen species (ROS) upon pathogen attack is generally considered a defense mechanism for microbial killing and an initiation of host defense responses in plants and animals. In this issue, Siddique et al. show that nicotinamide adenine dinucleotide phosphate oxidase-derived ROS function as a pathogenicity factor to promote the roundworm nematode infection in Arabidopsis thaliana, revealing the complex action of ROS in host-pathogen interactions.


Generation of whistler-wave heated discharges with planar resonant RF networks.
Magnetized plasma discharges generated by a planar resonant rf network are investigated. A regime transition is observed above a magnetic field threshold, associated with rf waves propagating in the plasma and which present the characteristics of whistler waves. These wave heated regimes can be considered as analogous to conventional helicon discharges, but in planar geometry.


Honeydew honey as a potent antibacterial agent in eradication of multi-drug resistant Stenotrophomonas maltophilia isolates from cancer patients.
Multi-drug resistance in nosocomial pathogens is a continually evolving and alarming problem in health care units. Since ancient times, honey has been used successfully for the treatment of a broad spectrum of infections with no risk of resistance development. This study investigated the antibacterial activity of two natural honeys, namely honeydew and manuka, against 20 nosocomial multi-drug resistant Stenotrophomonas maltophilia (S. maltophilia) isolates from cancer patients. An antibiotic susceptibility test was carried out using the disk diffusion method with 20 antibiotic disks. The antibacterial activity of honey was determined using a broth dilution method. The concentration of honey used in the study was within the range of 3.75% to 25% (w/v). All 20 clinical isolates were multi-drug resistant against 11 to 19 antibiotics. The MICs for honeydew honey ranged from 6.25% to 17.5%, while those for active manuka honey ranged from 7.5% to 22.5%. Honeydew honey had lower MICs than manuka honey against 16 of the tested isolates. This study showed that Slovak honeydew honey has exceptional antibacterial activity against multi-drug resistant S. maltophilia isolates and was more efficient than manuka honey (UMF 15+). Honeydew honey with strong antibacterial activity could be used as a potential agent to eradicate multi-drug resistant clinical isolates.


The clinical spectrum of hydrocephalus in adults: report of the first 517 patients of the Adult Hydrocephalus Clinical Research Network registry.
The authors describe the demographics and clinical characteristics of the first 517 patients enrolled in the Adult Hydrocephalus Clinical Research Network (AHCRN) during its first 2 years. Adults ≥ 18 years were nonconsecutively enrolled in a registry at 6 centers. Four categories of adult hydrocephalus were defined: transition (treated before age 18 years), unrecognized congenital (congenital pattern, not treated before age 18 years), acquired (secondary to known risk factors, treated or untreated), and suspected idiopathic normal pressure hydrocephalus (iNPH) (≥ age 65 years, not previously treated). Data include etiology, symptoms, examination findings, neuropsychology screening, comorbidities, treatment, complications, and outcomes. Standard evaluations were administered to all patients by trained examiners, including the Montreal Cognitive Assessment, the Symbol Digit Modalities Test, the Beck Depression Inventory-II, the Overactive Bladder Questionnaire Short Form symptom bother, the 10-Meter Walk Test, the Boon iNPH gait scale, the Lawton Activities of Daily Living/Instrumental Activities of Daily Living (ADL/IADL) questionnaire, the iNPH grading scale, and the modified Rankin Scale. Overall, 517 individuals were enrolled. Age ranged from 18.1 to 90.7 years, with patients in the transition group (32.7 ± 10.0 years) being the youngest and those in the suspected iNPH group (76.5 ± 5.2 years) being the oldest. The proportion of patients in each group was as follows: 16.6% transition, 26.5% unrecognized congenital, 18.2% acquired, and 38.7% suspected iNPH. Excluding the 86 patients in the transition group, who all had received treatment, 79.4% of adults in the remaining 3 groups had not been treated at the time of enrollment. Patients in the suspected iNPH group had the poorest performance in cognitive evaluations, and those in the unrecognized congenital group had the best performance. The same pattern was seen in the Lawton ADL/IADL scores. Gait velocity was lowest in patients in the suspected iNPH group. Categories that had the most comorbidities (suspected iNPH) or etiologies of hydrocephalus that directly cause neurological injury (transition, acquired) had greater degrees of impairment compared to unrecognized congenital, which had the fewest comorbidities or etiologies associated with neurological injury. The clinical spectrum of hydrocephalus in adults comprises more than iNPH or acquired hydrocephalus. Only 39% of patients had suspected iNPH, whereas 43% had childhood onset (i.e., those in the transition and unrecognized congenital groups). The severity of symptoms and impairment was worsened when the etiology of the hydrocephalus or complications of treatment caused additional neurological injury or when multiple comorbidities were present. However, more than half of patients in the transition, unrecognized congenital, and acquired hydrocephalus groups had minimal or no impairment. Excluding the transition group, nearly 80% of patients in the AHCRN registry were untreated at the time of enrollment. A future goal for the AHCRN is to determine whether patients with unrecognized congenital and acquired hydrocephalus need treatment and which patients in the suspected iNPH cohort actually have possible hydrocephalus and should undergo further diagnostic testing. Future prospective research is needed in the diagnosis, treatment, outcomes, quality of life, and macroeconomics of all categories of adult hydrocephalus.


Conducting a microbiome study.
Human microbiome research is an actively developing area of inquiry, with ramifications for our lifestyles, our interactions with microbes, and how we treat disease. Advances depend on carefully executed, controlled, and reproducible studies. Here, we provide a Primer for researchers from diverse disciplines interested in conducting microbiome research. We discuss factors to be considered in the design, execution, and data analysis of microbiome studies. These recommendations should help researchers to enter and contribute to this rapidly developing field.


Spatial Light Interference Microscopy (SLIM) using twisted-nematic liquid-crystal modulation.
We report the use of a twisted nematic liquid-crystal spatial light modulator (TNLC-SLM) for quantitative phase imaging. The experimental setup is a new implementation of the SLIM principle, which is a phase shifting, white light method for quantitative phase imaging. The approach is based on switching between the phase and amplitude modulation modes of the SLM. Our system is able to deliver a 0.99 nm spatial and 1.33 nm temporal pathlength sensitivity while retaining the optical transverse resolution. The system is implemented as an additional module mounted to a conventional microscope, which makes the system very easy to deploy and integrate with other imaging modalities.


Evaluation of some anthracycline antibiotics in an in vivo model for studying drug-induced human leukemia cell differentiation.
Human acute promyelocytic leukemia cells (HL-60) were induced to undergo terminal differentiation by treatment in vivo with marcellomycin. This was accomplished by cloning HL-60 cells in 0.3% agar in diffusion chambers, which were subsequently implanted in the peritoneal cavities of mice. These animals were given injections i.v. with anthracycline antibiotics, and the chambers were transferred to a second recipient 24 hr later. After an additional 8 days, the chambers were removed, the cloning efficiency was determined, and colonies were scored for the presence of differentiated cells based on their ability to reduce nitro blue tetrazolium (NBT). Marcellomycin produced dose-dependent decreases in cloning efficiency and increases in the number of differentiated cells present in chambers. At all dose levels of marcellomycin tested, three types of colonies were observed; these were colonies consisting entirely of undifferentiated cells and approximately equal numbers of colonies consisting of solely differentiated cells and those with mixtures of both differentiated and undifferentiated cells. An 8-fold increase in colonies containing differentiated cells (both pure and mixed) was observed after a single injection of marcellomycin (10 mg/kg), a dose which reduced cloning efficiency by 40%. At that dosage level, aclacinomycin A also induced differentiation, while doxorubicin was ineffective, a finding consistent with the effects of these anthracyclines on HL-60 cells in suspension culture. In addition to the functional changes accompanying differentiation, commitment was characterized by a limitation in proliferative potential. Thus, the average size of uniform NBT-positive colonies was approximately 16 cells, and few clones of NBT-positive cells greater than 32 cells in number were observed; this contrasted with NBT-negative clones which contained up to 100 cells. This finding suggests that HL-60 cells undergo an average of four and a maximum of five divisions upon commitment to granulocytic differentiation. The in vivo system described may be useful in further evaluation of differentiation-inducing agents for therapeutic potential after an initial in vitro screen to identify active compounds.


Max: a helix-loop-helix zipper protein that forms a sequence-specific DNA-binding complex with Myc.
The myc protooncogene family has been implicated in cell proliferation, differentiation, and neoplasia, but its mechanism of function at the molecular level is unknown. The carboxyl terminus of Myc family proteins contains a basic region helix-loop-helix leucine zipper motif (bHLH-Zip), which has DNA-binding activity and has been predicted to mediate protein-protein interactions. The bHLH-Zip region of c-Myc was used to screen a complementary DNA (cDNA) expression library, and a bHLH-Zip protein, termed Max, was identified. Max specifically associated with c-Myc, N-Myc, and L-Myc proteins, but not with a number of other bHLH, bZip, or bHLH-Zip proteins. The interaction between Max and c-Myc was dependent on the integrity of the c-Myc HLH-Zip domain, but not on the basic region or other sequences outside the domain. Furthermore, the Myc-Max complex bound to DNA in a sequence-specific manner under conditions where neither Max nor Myc exhibited appreciable binding. The DNA-binding activity of the complex was dependent on both the dimerization domain and the basic region of c-Myc. These results suggest that Myc family proteins undergo a restricted set of interactions in the cell and may belong to the more general class of eukaryotic DNA-binding transcription factors.


Increasing compliance with neuromuscular training to prevent ankle sprain in sport: does the 'Strengthen your ankle' mobile App make a difference? A randomised controlled trial.
E-health has the potential to facilitate implementation of effective measures to prevent sports injuries. We evaluated whether an interactive mobile application containing a proven effective exercise programme to prevent recurrent ankle sprains resulted in higher compliance as compared with regular written exercise materials. 220 athletes participated in this randomised controlled trial with a follow-up of 8 weeks; 110 athletes received a booklet explaining an 8-week neuromuscular training programme; 110 athletes participated in the same programme in an interactive mobile App (Strengthen your ankle). The primary outcome was compliance with the exercise programme. Secondary outcome measure was the incidence density of self-reported recurrent ankle sprains. The mean compliance to the exercise scheme was 73.3% (95% CI 67.7% to 78.1%) in the App group, compared with 76.7% (95% CI 71.9% to 82.3%) in the Booklet group. No significant difference in compliance was found between groups. The incidence densities of self-reported time-loss recurrences were not significantly different between both groups (HR 3.07; 95% CI 0.62 to 15.20). This study shows that the method of implementing the exercises by using an App or a Booklet does not lead to different compliance rates. The use of a mobile App or a Booklet lead to similar compliance and injury rates in the short term. The Netherlands National Trial Register NTR 4027. The NTR is part of the WHO Primary Registries.


Ping-pong amplification of a retroviral vector achieves high-level gene expression: human growth hormone production.
Retroviral vectors offer major advantages for gene transfer studies but have not been useful for producing proteins in large quantities. This deficiency has resulted in part from interference to superinfection, which limits the numbers of active proviruses in cells. Recently, we found that these vectors amplify when they are added as calcium phosphate precipitates to cocultures of cells that package retroviruses into ecotropic and amphotropic host range envelopes. Helper-free virions from either cell type can infect the other without interference, resulting in theoretically limitless back-and-forth (ping-pong) vector replication. In initial studies, however, amplifications of a vector that contained the human growth hormone gene ceased when the hormone produced was 0.3% or less of cellular protein synthesis. This limit was caused by two factors. First, recombinant shutoff viruses that are replication defective and encode envelope glycoproteins form at a low probability during any round of the vector replication cycle and these spread in cocultures, thereby establishing interference. Single cells in shutoff cocultures therefore synthesize both ecotropic and amphotropic envelope glycoproteins, and they release promiscuous (presumably hybrid) virions. The probability of forming shutoff viruses before the vector had amplified to a high multiplicity was reduced by using small cocultures. Second, cells with large numbers of proviruses are unhealthy and their proviral expression can be unstable. Stable expresser cell clones were obtained by selection. Thereby, cell lines were readily obtained that stably produce human growth hormone as 4 to 6% of the total protein synthesis. A ping-pong retroviral vector can be used for high-level protein production in vertebrate cells.