Is Resident-Driven Inpatient Care More Expensive? Challenging a Long-Held Assumption.
The financial impact of graduate medical education (GME) on teaching hospitals remains poorly understood, while calls for increased federal support continue alongside legislative threats to reduce funding. Despite studies suggesting that residents are more "economical" than alternative providers, GME is widely believed to be an expensive investment. Assumptions that residents increase the cost of patient care have persisted in the absence of convincing evidence to the contrary. Thus, the authors sought to examine resident influence on patient care costs by comparing costs between a resident-driven service (RS) and a nonresident-covered service (NRS), with attention to clinical outcomes and how potential cost differences relate to the utilization of resources, length of stay (LOS), and other factors. This prospective study compared costs and clinical outcomes of internal medicine patients admitted to an RS versus an NRS at Massachusetts General Hospital (July 1, 2016-June 30, 2017). Total variable direct costs of inpatient admission was the primary outcome measure. LOS; 30-day readmission rate; utilization related to diagnostic radiology, pharmaceuticals, and clinical labs; and other outcome measures were also compared. Linear regression models quantified the relationship between log-transformed variable direct costs and service. Baseline characteristics of 5,448 patients on the 2 services (3,250 on an RS and 2,198 on an NRS) were similar. On an RS, patient care costs were slightly less and LOS was slightly shorter than on an NRS, with no significant differences in hospital mortality or 30-day readmission rate detected. Resource utilization was comparable between the services. These findings undermine long-held assumptions that residents increase the cost of patient care. Though not generalizable to ambulatory settings or other specialties, this study can help inform hospital decision making around sponsorship of GME programs, especially if federal funding for GME remains capped or is subject to additional reductions.


Edge-Confined Excitons in Monolayer Black Phosphorus.
Quantum confinement of two-dimensional excitons in van der Waals materials via electrostatic trapping, lithographic patterning, Moiré potentials, and chemical implantation has enabled significant advances in tailoring light emission from nanostructures. While such approaches rely on complex preparation of materials, natural edges are a ubiquitous feature in layered materials and provide a different approach for investigating quantum-confined excitons. Here, we observe that certain edge sites of monolayer black phosphorus (BP) strongly localize the intrinsic quasi-one-dimensional excitons, yielding sharp spectral lines in photoluminescence, with nearly an order of magnitude line width reduction. Through structural characterization of BP edges using transmission electron microscopy and first-principles GW plus Bethe-Salpeter equation (GW-BSE) calculations of exemplary BP nanoribbons, we find that certain atomic reconstructions can strongly quantum-confine excitons resulting in distinct emission features, mediated by local strain and screening. We observe linearly polarized luminescence emission from edge reconstructions that preserve the mirror symmetry of the parent BP lattice, in agreement with calculations. Furthermore, we demonstrate efficient electrical switching of localized edge excitonic luminescence, whose sites act as excitonic transistors for emission. Localized emission from BP edges motivates exploration of nanoribbons and quantum dots as hosts for tunable narrowband light generation, with future potential to create atomic-like structures for quantum information processing applications as well as exploration of exotic phases that may reside in atomic edge structures.


Protocol for determination of phosphatidylinositol 3-phosphate levels and localization in Candida glabrata by confocal microscopy.
Phosphatidylinositol 3-phosphate (PI3P) levels govern membrane trafficking in Candida glabrata. Here, we present a confocal imaging-based protocol for PI3P localization analysis using the GFP-FYVE (found in Fab1, YOTB, Vac1, and EEA1) fusion protein. We describe steps for cloning the FYVE domain into the GFP-containing vector backbone, transforming FYVE-GFP into C. glabrata, and preparing slides with FYVE-GFP-expressing C. glabrata cells. We then detail procedures for acquiring and analyzing images and quantifying signal data. This protocol is adaptable to subcellular localization analysis of other low-abundant lipid and protein molecules. For complete details on the use and execution of this protocol, please refer to Askari et al. (2023).1.


Aldehyde dehydrogenase 1 (ALDH1) expression is an independent prognostic factor in triple negative breast cancer (TNBC).
Triple negative breast cancer (TNBC) is a subset of breast cancer that is highly aggressive and has a poor prognosis. Meanwhile, cancer stem cells (CSCs) are also characterized by a strong tumorigenic potential, which might be partly responsible for the aggressive behavior of TNBC. We previously showed that CSCs are enriched in TNBC cell lines and tissues. Further experiments in animal models revealed higher tumorigenicity of CSCs sorted from TNBC cell lines. In this study, we aimed to determine the clinical relationship between CSCs and TNBC by exploring the expression of aldehyde dehydrogenase 1 (ALDH1), which is a putative marker of breast CSCs, in TNBC tissues.ALDH1 levels in paraffin-embedded tumor tissues from 158 TNBC patients were evaluated by immunohistochemistry staining using an ALDH1A1 primary antibody. Staining evaluation was performed independently by two pathologists, and the expression level of ALDH1 was evaluated in terms of the percentage and intensity of positive cells. The association of immunohistochemistry staining of ALDH1 expression with clinical parameters was also analyzed.ALDH1 expression in tumor cells was observed in 88 out of 158 cases (55.7%). Analysis of clinicopathological parameters showed that the immunohistochemistry staining of ALDH1 was significantly correlated with tumor size (P = 0.02) and stage (P = 0.04). Survival analysis in patients with ALDH1 expression demonstrated shorter relapse-free survival (RFS) and overall survival (OS) times (P = 0.01; P = 0.001). Moreover, Cox multivariate analysis revealed that ALDH1 expression was an independent prognostic indicator of RFS and OS (P = 0.04; P = 0.04).Immunohistochemistry staining of ALDH1 in tumor cells is an independent prognostic indicator of RFS and OS in TNBC patients.


Neuropsychological similarities and differences among Huntington's disease, multiple sclerosis, and cortical dementia.
A comparison of cognitive function was made among patients with Huntington's disease, multiple sclerosis, and cortical dementia. Utilizing indexes from the Wechsler Adult Intelligence Scale and the Halstead-Reitan Battery, it was found that there was substantially more severe cognitive deficit in the Huntington's disease patients than in the multiple sclerosis patients, and the level of impairment was similar between the Huntington's disease and cortical dementia groups. Qualitative differences, particularly involving amount and type of perseveration, were noted among the three groups. It was concluded that subcortical dementia is not necessarily characterized by mild cognitive impairment, and there appear to be important qualitative differences between cortical and subcortical dementia. Results are discussed in terms of the usefulness of the presently conceptualized distinction between cortical and subcortical dementia.


Tumor Drug Penetration Measurements Could Be the Neglected Piece of the Personalized Cancer Treatment Puzzle.
Precision medicine aims to use patient genomic, epigenomic, specific drug dose, and other data to define disease patterns that may potentially lead to an improved treatment outcome. Personalized dosing regimens based on tumor drug penetration can play a critical role in this approach. State-of-the-art techniques to measure tumor drug penetration focus on systemic exposure, tissue penetration, cellular or molecular engagement, and expression of pharmacological activity. Using in silico methods, this information can be integrated to bridge the gap between the therapeutic regimen and the pharmacological link with clinical outcome. These methodologies are described, and challenges ahead are discussed. Supported by many examples, this review shows how the combination of these techniques provides enhanced patient-specific information on drug accessibility at the tumor tissue level, target binding, and downstream pharmacology. Our vision of how to apply tumor drug penetration measurements offers a roadmap for the clinical implementation of precision dosing.


Identification of germline susceptibility loci in ETV6-RUNX1-rearranged childhood acute lymphoblastic leukemia.
Acute lymphoblastic leukemia (ALL) is a malignant disease of the white blood cells. The etiology of ALL is believed to be multifactorial and likely to involve an interplay of environmental and genetic variables. We performed a genome-wide association study of 355 750 single-nucleotide polymorphisms (SNPs) in 474 controls and 419 childhood ALL cases characterized by a t(12;21)(p13;q22) - the most common chromosomal translocation observed in childhood ALL - which leads to an ETV6-RUNX1 gene fusion. The eight most strongly associated SNPs were followed-up in 951 ETV6-RUNX1-positive cases and 3061 controls from Germany/Austria and Italy, respectively. We identified a novel, genome-wide significant risk locus at 3q28 (TP63, rs17505102, P(CMH)=8.94 × 10(-9), OR=0.65). The separate analysis of the combined German/Austrian sample only, revealed additional genome-wide significant associations at 11q11 (OR8U8, rs1945213, P=9.14 × 10(-11), OR=0.69) and 8p21.3 (near INTS10, rs920590, P=6.12 × 10(-9), OR=1.36). These associations and another association at 11p11.2 (PTPRJ, rs3942852, P=4.95 × 10(-7), OR=0.72) remained significant in the German/Austrian replication panel after correction for multiple testing. Our findings demonstrate that germline genetic variation can specifically contribute to the risk of ETV6-RUNX1-positive childhood ALL. The identification of TP63 and PTPRJ as susceptibility genes emphasize the role of the TP53 gene family and the importance of proteins regulating cellular processes in connection with tumorigenesis.


Hemispheric differences in hemodynamics elicited by auditory oddball stimuli.
Evidence from neuroimaging studies suggests that the right hemisphere of the human brain might be more specialized for attention than the left hemisphere. However, differences between right and left hemisphere in the magnitude of hemodynamic activity (i.e., 'functional asymmetry') rarely have been explicitly examined in previous neuroimaging studies of attention. This study used a new voxel-based comparison method to examine hemispheric differences in the amplitude of the hemodynamic response in response to infrequent target, infrequent novel, and frequent standard stimuli during an event-related fMRI auditory oddball task in 100 healthy adult participants. Processing of low probability task-relevant target stimuli, or 'oddballs', and low probability task-irrelevant novel stimuli is believed to engage in orienting and attentional processes. It was hypothesized that greater right-hemisphere activation compared to left would be observed to infrequent target and novel stimuli. Consistent with predictions, greater right hemisphere than left frontal, temporal, and parietal lobe activity was observed for target detection and novelty processing. Moreover, asymmetry effects did not differ with respect to age or gender of the participants. The results (1) support the proposal that the right hemisphere is differentially engaged in processing salient stimuli and (2) demonstrate the successful use of a new voxel-based laterality analysis technique for fMRI data.


Special pharmacokinetics of fluconazole in septic, obese and burn patients.
Although there are many potential changes of pharmacokinetic parameters in patients with thermal injury, obesity or septicemia, data about the actual effect on pharmacokinetics and clinical efficacy of fluconazole are very limited. As current dosing recommendations are derived from healthy subjects and patients with normal weight, these recommendations may be inaccurate when applied to the patient populations mentioned above. Pharmacokinetic data of 14 patients with thermal injury were reviewed and revealed a shorter half life and more rapid clearance of fluconazole. In a subgroup of five patients, distribution volume was up to 2 times larger as usual with no relationship to creatinine clearance and degree of burns. In one extremely obese patient treated with fluconazole 1200 mg/day*, the corresponding mean steady-state plasma concentration and AUC were decreased with an increase of fluconazole clearance possibly due to a larger volume of distribution. In patients with septicemia, fluconazole plasma levels appear to be highly variable. As a considerable number of these patients develops acute renal failure, renal replacement therapy may be indicated which may require substantial dosage modifications of fluconazole.


The physiology of wandering behaviour in Manduca sexta. I. Temporal organization and the influence of the internal and external environments.
A stereotyped series of behavioural changes occurs in preparation for pupation in Manduca sexta. Feeding declines over an 8-h period, after which the larva coats its body with a labial gland secretion. The animal then begins a 10- to 30-h wandering behaviour during which it constructs a pupation chamber underground. Wandering behaviour starts during a specific temporal gate which is determined by an internal circadian timer. The scotophase of the day prior to wandering has the major influence on the timing of internal processes which activate the behaviour. Wandering duration is correlated with larval size, reflecting a possible influence of juvenile hormone. The larva appears to be irrevocably committed to begin wandering by an event that occurs about 15 h previously, a time that corresponds to the second of three prothoracicotropic hormone (PTTH) pulses and the accompanying elevation of ecdysteroids as measured by Gilbert et al. (1981). We conclude that both the initiation and duration of wandering behaviour are governed primarily by processes which are internal to the larva.


Appointment length, psychiatrists' communication behaviors, and medication management appointment adherence.
The authors explored the relationship between critical elements of medication management appointments (appointment length, patient-centered talk, and positive nonverbal affect among providers) and patient appointment adherence. The authors used an exploratory, cross-sectional design employing quantitative analysis of 83 unique audio recordings of split treatment medication management appointments for 46 African-American and 37 white patients with 24 psychiatrists at four ambulatory mental health clinics. All patients had a diagnosis of depression. Data collected included demographic information; Patient Health Questionnaire-9 scores for depression severity; psychiatrist verbal and nonverbal communication behaviors during medication management appointments, identified by the Roter Interaction Analysis System during analysis of audio recordings; and appointment adherence. Bivariate analyses were employed to identify covariates that might influence appointment adherence. Generalized estimating equations (GEEs) were employed to assess the relationship between appointment length, psychiatrist patient-centered talk, and positive voice tone ratings and patient appointment adherence, while adjusting for covariates and the clustering of observations within psychiatrists. Wald chi square analyses were used to test whether all or some variables significantly influenced appointment adherence. GEE revealed a significant relationship between positive voice tone ratings and appointment adherence (p=.03). Chi square analyses confirmed the hypothesis of a positive and significant relationship between appointment adherence and positive voice tone ratings (p=.03) but not longer visit length and more patient-centered communication. The nonverbal conveyance of positive affect was associated with greater adherence to medication management appointments by depressed patients. These findings potentially have important implications for communication skills training and adherence research.


Anaplastic Thyroid Carcinoma Mimicking Cervical Tuberculosis: A Case Report.
Anaplastic thyroid carcinoma is a rare and aggressive form of thyroid cancer that has a poor prognosis and a high mortality rate. It is characterized by rapid growth and invasion of nearby tissues. It typically presents as a rapidly growing goiter or nodule that is firm to the touch and firmly attached to the underlying structures. Case reports of unusual presentations of anaplastic thyroid carcinoma have been reported. The presentation of anaplastic thyroid carcinoma mimicking cervical tuberculosis is very unusual. We reported a case of a 65-year-old patient who had a left cervical swelling that had been evolving for 4 months, causing dysphagia. Initial imaging showed a necrotic mass in the left lobe of the thyroid, communicating with a second necrotic mass in the subcutaneous tissue that was fistulized to the skin and suggesting cervical tuberculosis. The mass was incised with pus and whitish material resembling caseous tuberculosis was discharged. Acid-fast bacilli (AFB) Polymerase chain reaction (PCR) was negative and biopsy revealed a nonspecific granulomatous lesion. Due to the growth of the mass and the presence of a permeation nodule, a second biopsy was performed, revealing anaplastic thyroid carcinoma. The patient was referred for radiochemotherapy due to tumor inoperability.


Factors influencing variation in investigations after a negative CT brain scan in suspected subarachnoid haemorrhage: a qualitative study.
Variation in the approach to the patient with a possible subarachnoid haemorrhage (SAH) has been previously documented. The purpose of this study was to identify factors that influence emergency physicians' decisions about diagnostic testing after a normal CT brain scan for ED patients with a headache suspicious of a SAH. We conducted an interview-based qualitative study informed by social constructionist theory. Fifteen emergency physicians from six EDs across Queensland, Australia, underwent individual face-to-face or telephone interviews. Content analysis was performed whereby transcripts were examined and coded independently by two co-investigators, who then jointly agreed on the influencing factors. Six categories of influencing factors were identified. Patient interaction was at the forefront of the identified factors. This shared decision-making process incorporated 'what the patient wants' but may be biased by how the clinician communicates the benefits and harms of the diagnostic options to the patient. Patient risk profile, practice evidence and guidelines were also important. Other influencing factors included experiential factors of the clinician, consultation with colleagues and external influences where practice location and work processes impose constraints on test ordering external to the preferences of the clinician or patient. The six categories were organised within a conceptual framework comprising four components: the context, the evidence, the experience and the decision. When clinicians are faced with a diagnostic challenge, such as the workup of a patient with suspected SAH, there are a number of influencing factors that can result in a variation in approach. These need to be considered in approaches to improve the appropriateness and consistency of medical care.


Determination of optimal cut-off values for waist circumferences used for the diagnosis of the metabolic syndrome among Batswana adults (ELS 32).
To date, no definitive waist circumference (WC) cut-off values for abdominal obesity (AO) have been established for sub-Saharan Africa, including Botswana. Therefore, the classification of AO among these populations is based on European values. For accurate diagnosis of the metabolic syndrome (MetS), cut-off values reflective of the population investigated must be used. The study was an attempt to determine optimal cut-off values for AO among Batswana adults. The receiver operating characteristic curve was used to determine the optimal cut-off values for predicting at least two other risk factors of the MetS. Data were used from a descriptive cross-sectional study employing a complex multi-stage cluster sampling. Demographic and anthropometric measurements (weight and height, waist and hip circumferences), blood pressure, and blood glucose, triglycerides, high-density lipoprotein cholesterol and total cholesterol levels were collected from 384 men and 416 women in Gaborone and the surrounding villages. The ability of waist circumference to predict at least two other risk factors of the MetS gave cut-off values of ≥ 91.0 cm (sensitivity 69.1% and specificity of 90.8%, area under the curve 0.85) for men and ≥ 82.3 cm (sensitivity of 88.6% and specificity of 58.9%, area under the curve of 0.76) for women. There is a difference between the cut-off values for Europeans with those determined for Batswana adults. Inconsistencies in cut-off values used have the potential for undesirable consequences for cardiovascular risk stratification and prioritisation of preventative strategies for AO and the MetS. The need to determine population-, ethnic- and gender-based cut-off values for AO for Batswana adults has never been more paramount.


[Pontoscolex corethrurus (Annelidae: Oligochaeta) soil quality indicator in Eucalyptus grandis (Myrtacea) sites with slash and burn management].
Soil burning has been used in agricultural and forestry systems as a fundamental technique to clean the land and add some nutrients to the soil. In addition, earthworms are known to promote various soil functions since they contribute to aeration and organic matter and nutrients availability to other soil organisms. This study evaluated the effects of tropical forest crops management with presence-absence of Eucalyptus grandis on earthworm population in Huimanquillo, Tabasco, Mexico. Three sites (average area of 1-1.5ha each) with different management conditions were considered for soil and earthworm sampling (two depths and six replicates): without vegetation (SV) and recent slash-burned (38 days), forest crops of five years of production of E. grandis (Euc), and secondary vegetation of 15 years (Acah). Soil physico-chemical properties (apparent density, humidity, texture, pH, Ntot, OM, P, K, cationic capacity) were also evaluated, and earthworms were collected at the end of the rainy season (august-october 2007). We found that the sites soil is an acrisol acid, with pH 3.0-4.5 in the first 30cm depth. Organic matter content (OM) and total nitrogen (Ntot) in the recently burned sites were significantly lower (6-8% y 0.19-0.22%, respectively) than in sites with vegetation (OM=9-11%; el Ntot=0.27-0.33%). Only one species (P. corethrurus) was found in all the sampled areas, where most of the individuals were at juvenile stage (80%). The highest densities and biomass were found in Euc. treatment (166.4ind/m2 y 36.8g/m2) followed by Acah (138.7ind/m2 y 19.1g/m2 respectively), while the SV treatment showed of about an 80% reduced earthworm populations when compared to other treatments. Even though 15 years have passed over the secondary vegetation (Acah) still some perturbations were observed as the low abundance of the oligochaeta group. We concluded that the management used to culture E. grandis produces negative effects over the abundance and diversity of earthworms and soil nutrient availability.


A Human Bi-specific Antibody against Zika Virus with High Therapeutic Potential.
Zika virus (ZIKV), a mosquito-borne flavivirus, causes devastating congenital birth defects. We isolated a human monoclonal antibody (mAb), ZKA190, that potently cross-neutralizes multi-lineage ZIKV strains. ZKA190 is highly effective in vivo in preventing morbidity and mortality of ZIKV-infected mice. NMR and cryo-electron microscopy show its binding to an exposed epitope on DIII of the E protein. ZKA190 Fab binds all 180 E protein copies, altering the virus quaternary arrangement and surface curvature. However, ZIKV escape mutants emerged in vitro and in vivo in the presence of ZKA190, as well as of other neutralizing mAbs. To counter this problem, we developed a bispecific antibody (FIT-1) comprising ZKA190 and a second mAb specific for DII of E protein. In addition to retaining high in vitro and in vivo potencies, FIT-1 robustly prevented viral escape, warranting its development as a ZIKV immunotherapy.


Symposium 2: Modern approaches to nutritional research challenges: Targeted and non-targeted approaches for metabolite profiling in nutritional research.
The present report discusses targeted and non-targeted approaches to monitor single nutrients and global metabolite profiles in nutritional research. Non-targeted approaches such as metabolomics allow for the global description of metabolites in a biological sample and combine an analytical platform with multivariate data analysis to visualise patterns between sample groups. In nutritional research metabolomics has generated much interest as it has the potential to identify changes to metabolic pathways induced by diet or single nutrients, to explore relationships between diet and disease and to discover biomarkers of diet and disease. Although still in its infancy, a number of studies applying this technology have been performed; for example, the first study in 2003 investigated isoflavone metabolism in females, while the most recent study has demonstrated changes to various metabolic pathways during a glucose tolerance test. As a relatively new technology metabolomics is faced with a number of limitations and challenges including the standardisation of study design and methodology and the need for careful consideration of data analysis, interpretation and identification. Targeted approaches are used to monitor single or multiple nutrient and/or metabolite status to obtain information on concentration, absorption, distribution, metabolism and elimination. Such applications are currently widespread in nutritional research and one example, using stable isotopes to monitor nutrient status, is discussed in more detail. These applications represent innovative approaches in nutritional research to investigate the role of both single nutrients and diet in health and disease.


Effects of gestation and birth weight on the growth and development of very low birthweight small for gestational age infants: a matched group comparison.
To investigate the effects of small for gestational age (SGA) in very low birthweight (VLBW) infants on growth and development until the fifth year of life. VLBW (< 1500 g) infants, selected from a prospective study, were classified as SGA (n = 115) on the basis of birth weight below the 10th percentile for gestational age and were compared with two groups of appropriate for gestational age (AGA) infants matched according to birth weight (AGA-BW; n = 115) or gestation at birth (AGA-GA; n = 115). Prenatal, perinatal, and postnatal risk factors were recorded, and duration and intensity of treatment were computed from daily assessments. Body weight, length, and head circumference were measured at birth, five and 20 months (corrected for prematurity), and at 56 months. General development was assessed at five and 20 months with the Griffiths scale of babies abilities, and cognitive development at 56 months with the Columbia mental maturity scales, a vocabulary (AWST) and language comprehension test (LSVTA). Significant group differences were found in complications (pregnancy, birth, and neonatal), parity, and multiple birth rate. The AGA-GA group showed most satisfactory growth up to 56 months, with both the AGA-BW and SGA groups lagging behind. The AGA-GA group also scored significantly more highly on all developmental and cognitive tests than the other groups. Developmental test results were similar for the SGA and AGA-BW groups at five and 20 months, but AGA-BW infants (lowest gestation) had lower scores on performance intelligence quotient and language comprehension at 56 months than the SGA group. When prenatal and neonatal complications, parity, and multiple birth were accounted for, group differences in growth remained, but differences in cognitive outcome disappeared after five months. Being underweight and with a short gestation (SGA and VLBW) leads to poor weight gain and head growth in infancy but does not result in poorer growth than in infants of the same birth weight but shorter gestation (AGA-BW) in the long term. SGA is related to early developmental delay and later language problems; however, neonatal complications may have a larger detrimental effect on long term cognitive development of VLBW infants than whether they are born SGA or AGA.


Time efficiency and quality of outcomes in a model-free digital workflow using digital impression immediately after implant placement: A double-blind self-controlled clinical trial.
To assess the clinical and laboratory time efficiency and quality of outcomes for posterior single implant crowns by means of a model-free digital workflow using digital impressions immediately after implant placement. Forty patients missing a single posterior tooth received implant therapy. For within-subject comparison, digital impressions were taken immediately after implant placement and conventional impressions after implant healing. Two monolithic zirconia crowns were fabricated using a laboratory-based CAD-CAM system. One crown was produced from the immediate digital impression and a model-free digital workflow (test group), and the second crown was produced from the conventional impression and a hybrid workflow (control group). Clinical and laboratory time was recorded. Quality of outcomes was evaluated double-blinded. A paired-sample t test was applied for statistical analysis. The total mean chairside time (impression and delivery) was 23.2 min (95%CI 22.2, 24.3) in the test group and 25.7 min (95%CI 24.4, 26.9) in the control group (p = 0.013). Significantly less laboratory time was needed in the model-free digital workflow (13.6 min, 95%CI 11.5, 15.6) as compared to the model-based hybrid workflow (29.9 min, 95%CI 25.7, 34.2) (p < 0.05). At crown delivery, 4/40 (test) and 12/40 (control) had no need of chairside adjustments, and 6/40 (test) and 5/40 (control) implant crowns were in need of additional laboratory interventions. The fabrication of posterior single implant crowns using digital impressions taken immediately after implant placement and a model-free, laboratory-based digital workflow was more time efficient and resulted in similar quality of outcomes as a hybrid workflow using conventional impressions.


Bioinspired catalyst design and artificial metalloenzymes.
Many bioinspired transition-metal catalysts have been developed over the recent years. In this review the progress in the design and application of ligand systems based on peptides and DNA and the development of artificial metalloenzymes are reviewed with a particular emphasis on the combination of phosphane ligands with powerful molecular recognition and shape selectivity of biomolecules. The various approaches for the assembly of these catalytic systems will be highlighted, and the possibilities that the use of the building blocks of Nature provide for catalyst optimisation strategies are discussed.


Blocking the BKCa channel induces NF-κB nuclear translocation by increasing nuclear calcium concentration†.
Nuclear factor kappa B (NF-κB) transcriptionally regulates several genes involved in initiating uterine contractions. A key factor controlling NF-κB activity is its translocation to the nucleus. In myometrial smooth muscle cells (MSMCs), this translocation can be stimulated by the inflammatory molecule lipopolysaccharide (LPS) or by blocking the potassium calcium-activated channel subfamily M alpha 1 (KCNMA1 or BKCa) with paxilline (PAX). Here, we sought to determine the mechanism by which blocking BKCa causes NF-κB-p65 translocation to the nucleus in MSMCs. We show that LPS- and PAX-induced NF-κB-p65 translocation are similar in that neither depends on several mitogen-activated protein kinase pathways, but both require increased intracellular calcium (Ca2+). However, the nuclear transport inhibitor wheat germ agglutinin prevented NF-κB-p65 nuclear translocation in response to LPS but not in response to PAX. Blocking BKCa located on the plasma membrane resulted in a transient NF-κB-p65 nuclear translocation that was not sufficient to induce expression of its transcriptional target, suggesting a role for intracellular BKCa. We report that BKCa also localizes to the nucleus and that blocking nuclear BKCa results in an increase in nuclear Ca2+ in MSMCs. Together, these data suggest that BKCa localized on the nuclear membrane plays a key role in regulating nuclear Ca2+ and NF-κB-p65 nuclear translocation in MSMCs.


[Effects of chronic intermittent hypoxia on regulation of miRNA-214 in myocardial apoptosis in rats].
To observe the effects of chronic intermittent hypoxia (CIH) on myocardial cell apoptosis and explore the mechanism of microRNA-214 (miRNA-214) expression for myocardial cell apoptosis and cardiac dysfunction. A total of 20 adult male Sprague-Dawley rats were randomized into two groups (n = 10 each): for CIH group, the rats were raised in a CIH chamber 8 h daily for 6 weeks; for normal control (NC) group, the animal were exposed to normoxic air 7 h daily for 6 weeks. Hemodynamic values and myocardial function were measured via a cannula inserted into right common carotid artery. Myocardial cell apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method. The expressions of miRNA-214 and hypoxia-inducible factor-1α (HIF-1α) mRNA were observed by real-time polymerase chain reaction (PCR). The expressions of such apoptosis-related proteins as fatty acid synthase (Fas), caspase 3, caspase 8, B-cell lymphoma-2 (Bcl-2) and Bcl-2-Associated X Protein (Bax) were detected by Western blot. Compared with NC group, left ventricle end diastolic pressure (LVEDP) ((4.1 ± 0.5) vs (2.7 ± 0.4) mmHg, 1 mmHg = 0.133 kPa) increased, maximal rate of pressure decline (-dp/dtmax) ((3 005 ± 86) vs (5 918 ± 112) mmHg/s), maximal rate of pressure development (+dp/dtmax) ((4 118 ± 76) vs (7 547 ± 271) mmHg/s) and left ventricular systolic pressure (LVSP) ((90.7 ± 2.5) vs (132.7 ± 5.5) mmHg] decreased in CIH group (all P < 0.001). There were significantly more TUNEL positive cells in CIH group versus NC group (P < 0.001). The expression of miRNA-214 was significantly lower in CIH group than that in NC group (P < 0.001). In addition, HIF-1α mRNA level was significantly higher in CIH group than that in NC group (P < 0.001). The expressions of caspase 3, caspase 8, Fas and Bax increased significantly in CIH group versus NC group (all P < 0.001), suggesting increased apoptosis of myocardial cells in CIH group. Compared with NC group, the expression of Bcl-2 decreased significantly in CIH group (P < 0.001). CIH may decrease the hemodynamic values and myocardial function in rats. The expression of miRNA-214 in CIH group is significantly depressed through accelerated apoptosis of myocardial cells.


Subdural hematoma caused by epithelioid angiosarcoma originating from the skull.
Cancers metastatic to the skull or dura may cause subdural hematoma (SDH). However, the frequency is low, and the presence of underlying cancers has almost always been known in such situations. We report a case of skull angiosarcoma manifesting as SDH, posing a diagnostic challenge to physicians. A 75-year-old man visited our clinic with sensorimotor disturbance of gradual onset approximately 1 month after a minor head trauma. He was diagnosed with SDH after imaging studies, and underwent surgery to evacuate the hematoma. Because the hematoma was organized, surgery was switched from burr-hole drainage to craniotomy. The bone flap as well as the dura over the hematoma had grossly normal appearance, and only the hematoma itself was submitted for histological examination. Although postoperative recovery was uneventful, the patient experienced recurrence of the SDH 2 months after surgery. At the second surgery, the bone flap and dura were intermingled with tumor tissue, and histological examination revealed that an epithelioid angiosarcoma originating from the skull was responsible for the SDH. Timely diagnosis of angiosarcoma manifesting as SDH is difficult because of its rarity. In retrospect, however, the diagnosis might have been established earlier if the bone flap and/or the dura had been biopsied at the time of the first surgery. The present case gives us a lesson that SDH may be an unusual manifestation of malignant tumors of skull or dural origin, and histological examination of not only hematoma capsule but also of the surrounding tissues may provide important diagnostic clues.


Similarities and differences between on-scalp and conventional in-helmet magnetoencephalography recordings.
The development of new magnetic sensor technologies that promise sensitivities approaching that of conventional MEG technology while operating at far lower operating temperatures has catalysed the growing field of on-scalp MEG. The feasibility of on-scalp MEG has been demonstrated via benchmarking of new sensor technologies performing neuromagnetic recordings in close proximity to the head surface against state-of-the-art in-helmet MEG sensor technology. However, earlier work has provided little information about how these two approaches compare, or about the reliability of observed differences. Herein, we present such a comparison, based on recordings of the N20m component of the somatosensory evoked field as elicited by electric median nerve stimulation. As expected from the proximity differences between the on-scalp and in-helmet sensors, the magnitude of the N20m activation as recorded with the on-scalp sensor was higher than that of the in-helmet sensors. The dipole pattern of the on-scalp recordings was also more spatially confined than that of the conventional recordings. Our results furthermore revealed unexpected temporal differences in the peak of the N20m component. An analysis protocol was therefore developed for assessing the reliability of this observed difference. We used this protocol to examine our findings in terms of differences in sensor sensitivity between the two types of MEG recordings. The measurements and subsequent analysis raised attention to the fact that great care has to be taken in measuring the field close to the zero-line crossing of the dipolar field, since it is heavily dependent on the orientation of sensors. Taken together, our findings provide reliable evidence that on-scalp and in-helmet sensors measure neural sources in mostly similar ways.


Immunofluorescence localization of Z-DNA in chromosomes: quantitation by scanning microphotometry and computer-assisted image analysis.
Anti-Z-DNA polyclonal and monoclonal immunoglobulins raised against left-handed polynucleotides show various degrees of specificity for base sequence and substitution. Class 1 IgGs recognize all Z-DNA with equal affinity; class 2 IgGs show a preference for d(G-C)n sequences and class 3 IgGs for d(G-C)n sequences with substitutions at the C5 position of the pyrimidine. These antibodies served as probes for the localization of Z-DNA in polytene and metaphase chromosomes and in interphase chromatin by indirect immunofluorescence. A quantitative assessment of the binding of anti-Z-DNA IgGs to polytene chromosomes of Chironomus and Drosophila was made by scanning microphotometry and by computer-assisted image analysis of double immunofluorescence and DNA-specific dye fluorescence images. The three classes of antibodies bind to most of the bands in acid fixed polytene chromosomes of C. thummi; however, preferential binding of one class of antibody over another can be observed in certain regions. These differences can be quantitated by arithmetic division or subtraction of the normalized digital images. If a class 2 antibody is first bound at saturating concentrations the binding of class 1 antibody is reduced throughout most bands by 40-50%. However, the telomeres of the three large chromosomes bind greater than 10 times as much class 1 antibody as class 2 antibody, indicating that the Z-DNA tracts in these regions are comprised largely of alternating sequences containing the A X T basepair, e.g., A-C. High-resolution image analysis of class 1 and class 2 immunofluorescence patterns and the total DNA distribution from polytene chromosomes of D. melanogaster show that the two antibody distributions are very similar in a large majority of the bands, but they often deviate from the mean DNA distribution profile. Z-DNA sequences of both G-C and A-C type are detectable at all levels of ploidy from 2n to 2(13)n and in species as diverse as insects and man. We conclude that the vast majority of polytene chromosome bands (genes) contain one or a few DNA sequences with potential for undergoing the B----Z transition and contain both alternating purine-pyrimidine G-C and A-C tracts or mixed sequences. Highly heterochromatic bands and telomeres have more Z potential sequences than do other bands.


Repurposing a novel anti-cancer RXR agonist to attenuate murine acute GVHD and maintain graft-versus-leukemia responses.
The nuclear receptor (NR) subclass, retinoid X receptors (RXRs), exert immunomodulatory functions that control inflammation and metabolism via homodimers and heterodimers, with several other NRs, including retinoic acid receptors. IRX4204 is a novel, highly specific RXR agonist in clinical trials that potently and selectively activates RXR homodimers, but not heterodimers. In this study, in vivo IRX4204 compared favorably with FK506 in abrogating acute graft-versus-host disease (GVHD), which was associated with inhibiting allogeneic donor T-cell proliferation, reducing T-helper 1 differentiation, and promoting regulatory T-cell (Treg) generation. Recipient IRX4204 treatment reduced intestinal injury and decreased IFN-γ and TNF-α serum levels. Transcriptional analysis of donor T cells isolated from intestines of GVHD mice treated with IRX4204 revealed significant decreases in transcripts regulating proinflammatory pathways. In vitro, inducible Treg differentiation from naive CD4+ T cells was enhanced by IRX4204. In vivo, IRX4204 increased the conversion of donor Foxp3- T cells into peripheral Foxp3+ Tregs in GVHD mice. Using Foxp3 lineage-tracer mice in which both the origin and current FoxP3 expression of Tregs can be tracked, we demonstrated that IRX4204 supports Treg stability. Despite favoring Tregs and reducing Th1 differentiation, IRX4204-treated recipients maintained graft-versus-leukemia responses against both leukemia and lymphoma cells. Notably, IRX4204 reduced in vitro human T-cell proliferation and enhanced Treg generation in mixed lymphocyte reaction cultures. Collectively, these beneficial effects indicate that targeting RXRs with IRX4204 could be a novel approach to preventing acute GVHD in the clinic.


An Efficient Feature Selection Strategy Based on Multiple Support Vector Machine Technology with Gene Expression Data.
The application of gene expression data to the diagnosis and classification of cancer has become a hot issue in the field of cancer classification. Gene expression data usually contains a large number of tumor-free data and has the characteristics of high dimensions. In order to select determinant genes related to breast cancer from the initial gene expression data, we propose a new feature selection method, namely, support vector machine based on recursive feature elimination and parameter optimization (SVM-RFE-PO). The grid search (GS) algorithm, the particle swarm optimization (PSO) algorithm, and the genetic algorithm (GA) are applied to search the optimal parameters in the feature selection process. Herein, the new feature selection method contains three kinds of algorithms: support vector machine based on recursive feature elimination and grid search (SVM-RFE-GS), support vector machine based on recursive feature elimination and particle swarm optimization (SVM-RFE-PSO), and support vector machine based on recursive feature elimination and genetic algorithm (SVM-RFE-GA). Then the selected optimal feature subsets are used to train the SVM classifier for cancer classification. We also use random forest feature selection (RFFS), random forest feature selection and grid search (RFFS-GS), and minimal redundancy maximal relevance (mRMR) algorithm as feature selection methods to compare the effects of the SVM-RFE-PO algorithm. The results showed that the feature subset obtained by feature selection using SVM-RFE-PSO algorithm results has a better prediction performance of Area Under Curve (AUC) in the testing data set. This algorithm not only is time-saving, but also is capable of extracting more representative and useful genes.


A systematic review of lower extremity electrical stimulation for treatment of walking impairment in peripheral artery disease.
Lower extremity peripheral artery disease (PAD) induces an ischemic pain in the lower limbs and leads to walking impairment. Electrical stimulation has been used in patients with PAD, but no systematic review has been proposed to address the efficacy of the technique as a treatment for walking impairment in PAD. A systematic search was performed to identify trials focused on electrical stimulation for the treatment of walking impairment in patients with PAD in the Cochrane Central Register, PubMed, Embase, and the Web of Science. Studies were included where the primary outcomes were pain-free walking distance and/or maximal walking distance. When appropriate, eligible studies were independently assessed for quality using the Cochrane Collaboration's tool for assessing risk of bias. Five studies eligible for inclusion were identified, of which only two were randomized controlled studies. Trial heterogeneity prevented the use of the GRADE system and the implementation of a meta-analysis. Three types of electrical stimulation have been used: neuromuscular electrical stimulation (NMES, n = 3), transcutaneous electrical stimulation (n = 1), and functional electrical stimulation (n = 1). The two available randomized controlled studies reported a significant improvement in maximal walking distance (+40 m/+34% and +39 m/+35%, respectively) following a program of NMES. Owing to the low number of eligible studies, small sample size, and the risk of bias, no clear clinical indication can be drawn regarding the efficacy of electrical stimulation for the management of impaired walking function in patients with PAD. Future high-quality studies are required to define objectively the effect of electrical stimulation on walking capacity.


Endoglycosidase H-sensitive glycopeptides in eleven different animal cells.
We have examined the distribution of mannose-labelled glycopeptides in eleven different animal cells grown in vitro. All of the cells examined contained endoglycosidase H-sensitive species of high and low molecular weight, associated with the cell material and with the cell surface; however, the distribution between the two pools was different, suggesting a 'sorting out' of glycoproteins. Another conclusion from our studies is that the oligosaccharide processing known to occur during or after membrane glycoprotein translation is incomplete in a high percentage of mannose-containing N-linked oligosaccharides of cell surfaces. There was no consistent correlation between the relative amounts of endoglycosidase H-sensitive and -resistant glycopeptides and whether the cells were normal, virus-transformed or tumour-derived.


Moisture-Assisted Hydroboration of Nitriles and Conversion Thereof to N-Heterocyles and N-Containing Derivatives.
The recent revelation of hidden-borane catalysis has revolutionized the field of catalytic hydroboration in organic synthesis. Many nucleophilic reaction promoters, previously believed to be the catalysts, in fact primarily facilitated the formation of borane (BH3), which subsequently acted as the true catalyst. This revelation prompted us to explore the untapped potential of these unexpected transformations, with a view to simplify hydroboration using more cost-effective and environmentally friendly nucleophilic precatalysts. Via computational studies, we were able to identify that water can actually undertake that role. Herein, we report a study on the simple hydroboration of nitriles, a notoriously challenging yet synthetically valuable class of substrates, using nothing more than moisture as an activating agent. This moisture-assisted nitrile hydroboration process can seamlessly integrate with a range of downstream transformations in a one-pot fashion to produce valuable N-containing products such as symmetrical imines, thioureas, and bis(alcohol)amines as well as N-heterocycles such as pyrroles, pyridines, pyridinium salts, 2-iminothiazolines, and carbazoles.


Re-growth of an incomplete discoid lateral meniscus after arthroscopic partial resection in an 11 year-old boy: a case report.
Discoid lateral meniscus is common in children. Arthroscopic partial resection is indicated in symptomatic cases generally achieving satisfactory results. We present a case of an incomplete discoid lateral meniscus of the right knee in an 11 year-old boy, treated with arthroscopic partial resection, which developed a re-growth of the remnant, restoring the pre-operative incomplete discoid shape. To the best of our knowledge this is the first report about re-growth of a discoid meniscus after surgery. Debate still exists regarding the etiology of a discoid meniscus. Some authors proposed it is the persistence of the normal stage during fetal development. However, most other authors believe it is anomalous and arises through variant morphogenesis. The re-growth of the discoid lateral meniscus following surgery in this patient seems to prove this latter theory. The residual growth of the knee involves also the lateral meniscus and that may have contributed to restoring the meniscus to the previous condition. This case report demonstrates discoid meniscal re-growth in a child. The growth spurt may have an impact on meniscal regeneration. Re-growth of the discoid lateral meniscus in our patient favors the hypothesis of variant morphogenesis.


Selection of transposon mutants of Mycobacterium tuberculosis with increased macrophage infectivity identifies fadD23 to be involved in sulfolipid production and association with macrophages.
Alterations to the composition or architecture of the mycobacterial cell envelope can affect the macrophage infectivity of the bacillus. To further characterize the mycobacterial gene products that modulate the interaction with host cells, we employed transposon mutagenesis and screened for mutants that demonstrated an enhanced binding affinity toward macrophages. After successive rounds of mutant selection and enrichment, a total of five mutants were isolated that harboured gene disruptions within loci involved in lipid synthetic pathways as well as genes coding for putative hypothetical proteins. One mutant in particular, with a disruption in the Rv3826 gene (fadD23), was repeatedly isolated during library screening. Analysis of the cell envelope constituents of the Tn : : fadD23 strain revealed a lack of sulfolipid production which was restored following complementation with the wild-type gene.


[Primary diffuse leptomeningeal melanoblastosis and its CT appearances (author's transl)].
We report the case of a twelve-year-old boy who died, after a six months history, from primary diffuse leptomeningeal melanoblastosis, CT investigations showed a saw-tooth contrast enhancement at the brain surface. CT investigations may therefore be very helpful in meningeal lesions - apart from the importance of CSF investigations.


A 12-month follow-up of the immune response to SARS-CoV-2 primary vaccination: evidence from a real-world study.
A real-world population-based longitudinal study, aimed at determining the magnitude and duration of immunity induced by different types of vaccines against COVID-19, started in 2021 by enrolling a cohort of 2,497 individuals at time of their first vaccination. The study cohort included both healthy adults aged ≤65 years and elderly subjects aged >65 years with two or more co-morbidities. Here, patterns of anti-SARS-CoV-2 humoral and cell-mediated specific immune response, assessed on 1,182 remaining subjects, at 6 (T6) and 12 months (T12) after the first vaccine dose, are described. At T12 median anti-Spike IgG antibody levels were increased compared to T6. The determinants of increased anti-Spike IgG were the receipt of a third vaccine dose between T6 and T12 and being positive for anti-Nucleocapside IgG at T12, a marker of recent infection, while age had no significant effect. The capacity of T12 sera to neutralize in vitro the ancestral B strain and the Omicron BA.5 variant was assessed in a subgroup of vaccinated subjects. A correlation between anti-S IgG levels and sera neutralizing capacity was identified and higher neutralizing capacity was evident in healthy adults compared to frail elderly subjects and in those who were positive for anti-Nucleocapside IgG at T12. Remarkably, one third of T12 sera from anti-Nucleocapside IgG negative older individuals were unable to neutralize the BA.5 variant strain. Finally, the evaluation of T-cell mediated immunity showed that most analysed subjects, independently from age and comorbidity, displayed Spike-specific responses with a high degree of polyfunctionality, especially in the CD8 compartment. In conclusion, vaccinated subjects had high levels of circulating antibodies against SARS-CoV-2 Spike protein 12 months after the primary vaccination, which increased as compared to T6. The enhancing effect could be attributable to the administration of a third vaccine dose but also to the occurrence of breakthrough infection. Older individuals, especially those who were anti-Nucleocapside IgG negative, displayed an impaired capacity to neutralize the BA.5 variant strain. Spike specific T-cell responses, able to sustain immunity and maintain the ability to fight the infection, were present in most of older and younger subjects assayed at T12.


What influences chronic pain management? A best-worst scaling experiment with final year medical students and general practitioners.
Chronic pain education is an essential determinant for optimal chronic pain management. Given that attitudes and preferences are involved in making treatment decisions, identifying which factors are most influential to final year medical students' and general practitioners' (GPs) chronic pain management choices is of importance. This study investigates Swedish and Australian students' preferences with respect to a chronic pain condition, using a best-worst scaling (BWS) experiment, which is designed to rank alternatives. BWS, a stated-preference method grounded in random utility theory, was used to explore the importance of factors influencing chronic pain management. All three cohorts considered the patients' pain description and previous treatment experience as the most important factors in making treatment decisions, whereas their demographics and voices or facial expressions while describing their pain were considered least important. Factors such as social support, patient preferences and treatment adherence were, however, disregarded by all cohorts in favour of pain assessment factors such as pain ratings, description and history. Swedish medical students and GPs show very high correlation in their choices, although the GPs consider their professional experience as more important compared to the students. This study suggests that the relative importance of treatment factors is cemented early and thus underline the critical importance of improving pain curricula during undergraduate medical education.


Antioxidants and free radical scavengers for the treatment of stroke, traumatic brain injury and aging.
The overproduction of reactive oxygen species (ROS) and reactive nitrogen species (RNS) is a common underlying mechanism of many neuropathologies, as they have been shown to damage various cellular components, including proteins, lipids and DNA. Free radicals, especially superoxide (O(2)*-), and non-radicals, such as hydrogen peroxide (H(2)O(2)), can be generated in quantities large enough to overwhelm endogenous protective enzyme systems, such as superoxide dismutase (SOD) and reduced glutathione (GSH). Here we review the mechanisms of ROS and RNS production, and their roles in ischemia, traumatic brain injury and aging. In particular, we discuss several acute and chronic pharmacological therapies that have been extensively studied in order to reduce ROS/RNS loads in cells and the subsequent oxidative stress, so-called "free-radical scavengers." Although the overall aim has been to counteract the detrimental effects of ROS/RNS in these pathologies, success has been limited, especially in human clinical studies. This review highlights some of the recent successes and failures in animal and human studies by attempting to link a compound's chemical structure with its efficacy as a free radical scavenger. In particular, we demonstrate how antioxidants derived from natural products, as well as long-term dietary alterations, may prove to be effective scavengers of ROS and RNS.


Perisomatic changes in h-channels regulate depressive behaviors following chronic unpredictable stress.
Chronic stress can be a precipitating factor in the onset of depression. Lentiviral-mediated knockdown of HCN1 protein expression and reduction of functional Ih produce antidepressant behavior. However, whether h-channels are altered in an animal model of depression is not known. We found that perisomatic HCN1 protein expression and Ih-sensitive physiological measurements were significantly increased in dorsal but not in ventral CA1 region/neurons following chronic unpredictable stress (CUS), a widely accepted model for major depressive disorder. Cell-attached patch clamp recordings confirmed that perisomatic Ih was increased in dorsal CA1 neurons following CUS. Furthermore, when dorsal CA1 Ih was reduced by shRNA-HCN1, the CUS-induced behavioral deficits were prevented. Finally, rats infused in the dorsal CA1 region with thapsigargin, an irreversible inhibitor of the SERCA pump, exhibited anxiogenic-like behaviors and increased Ih, similar to that observed following CUS. Our results suggest that CUS, but not acute stress, leads to an increase in perisomatic Ih in dorsal CA1 neurons and that HCN channels represent a potential target for the treatment of major depressive disorder.


Functionally important amino acids in the Arabidopsis thylakoid phosphate transporter: homology modeling and site-directed mutagenesis.
The anion transporter 1 (ANTR1) from Arabidopsis thaliana, homologous to the mammalian members of the solute carrier 17 (SLC17) family, is located in the chloroplast thylakoid membrane. When expressed heterologously in Escherichia coli, ANTR1 mediates a Na(+)-dependent active transport of inorganic phosphate (P(i)). The aim of this study was to identify amino acid residues involved in P(i) binding and translocation by ANTR1 and in the Na(+) dependence of its activity. A three-dimensional structural model of ANTR1 was constructed using the crystal structure of glycerol 3-phosphate/phosphate antiporter from E. coli as a template. Based on this model and multiple sequence alignments, five highly conserved residues in plant ANTRs and mammalian SLC17 homologues have been selected for site-directed mutagenesis, namely, Arg-120, Ser-124, and Arg-201 inside the putative translocation pathway and Arg-228 and Asp-382 exposed at the cytoplasmic surface of the protein. The activities of the wild-type and mutant proteins have been analyzed using expression in E. coli and radioactive P(i) transport assays and compared with bacterial cells carrying an empty plasmid. The results from P(i)- and Na(+)-dependent kinetics indicate the following: (i) Arg-120 and Arg-201 may be important for binding and translocation of the substrate; (ii) Ser-124 may function as a transient binding site for Na(+) ions in close proximity to the periplasmic side; (iii) Arg-228 and Asp-382 may participate in interactions associated with protein conformational changes required for full transport activity. Functional characterization of ANTR1 should provide useful insights into the function of other plant and mammalian SLC17 homologous transporters.


Lessons learned about pneumonic plague diagnosis from two outbreaks, Democratic Republic of the Congo.
Pneumonic plague is a highly transmissible infectious disease for which fatality rates can be high if untreated; it is considered extremely lethal. Without prompt diagnosis and treatment, disease management can be problematic. In the Democratic Republic of the Congo, 2 outbreaks of pneumonic plague occurred during 2005 and 2006. In 2005, because of limitations in laboratory capabilities, etiology was confirmed only through retrospective serologic studies. This prompted modifications in diagnostic strategies, resulting in isolation of Yersinia pestis during the second outbreak. Results from these outbreaks demonstrate the utility of a rapid diagnostic test detecting F1 antigen for initial diagnosis and public health management, as well as the need for specialized sampling kits and trained personnel for quality specimen collection and appropriate specimen handling and preservation for plague confirmation and Y. pestis isolation. Efficient frontline management and a streamlined diagnostic strategy are essential for confirming plague, especially in remote areas.


Cytomegalovirus activates interferon immediate-early response gene expression and an interferon regulatory factor 3-containing interferon-stimulated response element-binding complex.
Interferon establishes an antiviral state in numerous cell types through the induction of a set of immediate-early response genes. Activation of these genes is mediated by phosphorylation of latent transcription factors of the STAT family. We found that infection of primary foreskin fibroblasts with human cytomegalovirus (HCMV) causes selective transcriptional activation of the alpha/beta-interferon-responsive ISG54 gene. However, no activation or nuclear translocation of STAT proteins was detected. Activation of ISG54 occurs independent of protein synthesis but is prevented by protein tyrosine kinase inhibitors. Further analysis revealed that HCMV infection induced the DNA binding of a novel complex, tentatively called cytomegalovirus-induced interferon-stimulated response element binding factor (CIF). CIF is composed, at least in part, of the recently identified interferon regulatory factor 3 (IRF3), but it does not contain the STAT1 and STAT2 proteins that participate in the formation of interferon-stimulated gene factor 3. IRF3, which has previously been shown to possess no intrinsic transcriptional activation potential, interacts with the transcriptional coactivator CREB binding protein, but not with p300, to form CIF. Activating interferon-stimulated genes without the need for prior synthesis of interferons might provide the host cell with a potential shortcut in the activation of its antiviral defense.


A qualitative study to assess perceptions, barriers, and motivators supporting smokeless tobacco cessation in the US fire service.
While firefighters currently have low smoking rates, rates of smokeless tobacco (SLT) use among this population are remarkably high and substantially greater than similar occupational groups, and the general population. This study explored determinants associated with SLT use, barriers to cessation, and motivators for SLT cessation in the fire service. Key informant interviews were conducted in 23 career firefighters who were current (n = 14) and former (n = 9) SLT users from across the U.S. Discussions were recorded and independently coded according to questions and themes. Major themes that developed among firefighters regarding SLT use determinants included positive perceptions of SLT products, social influences from their peers and family members, acceptability of SLT use in the fire service, and a coping resource for job stress. Firefighters discussed several barriers to SLT cessation, including intrapersonal barriers such as SLT use habits and its dependency, concerns about withdrawal symptoms; and social-environmental barriers including lack of support from health and other services providers, and lack of enforcement of existing tobacco policies regarding SLT use. Firefighters also mentioned both internal and external motivators for cessation. Internal motivators included self-motivation and their health concerns while external motivators included friends and family support, incentives or rewards, and price of SLT products. Findings provide unique perspectives from firefighters on factors that influence SLT use and barriers and motivators to SLT cessation. These are insufficiently assessed and considered by the fire service organizations and their health care providers. Thus, the organizations must understand these issues in order to mitigate barriers and motivate the personnel to quit using SLT. Information gained from firefighters who were current and former SLT users can be used to develop an effective, culturally-tailored intervention that is acceptable to fire service personnel.


Cardiac reprogramming reduces inflammatory macrophages and improves cardiac function in chronic myocardial infarction.
Cardiomyocytes (CMs) have little regenerative capacity. After myocardial infarction (MI), scar formation and myocardial remodeling proceed in the infarct and non-infarct areas, respectively, leading to heart failure (HF). Prolonged activation of cardiac fibroblasts (CFs) and inflammatory cells may contribute to this process; however, therapies targeting these cell types remain lacking. Cardiac reprogramming converts CFs into induced CMs, reduces fibrosis, and improves cardiac function in chronic MI through the overexpression of Mef2c/Gata4/Tbx5/Hand2 (MGTH). However, whether cardiac reprogramming reduces inflammation in infarcted hearts remains unclear. Moreover, the mechanism through which MGTH overexpression in CFs affects inflammatory cells remains unknown. Here, we showed that inflammation persists in the myocardium until three months after MI, which can be reversed with cardiac reprogramming. Single-cell RNA sequencing demonstrated that CFs expressed pro-inflammatory genes and exhibited strong intercellular communication with inflammatory cells, including macrophages, in chronic MI. Cardiac reprogramming suppressed the inflammatory profiles of CFs and reduced the relative ratios and pro-inflammatory signatures of cardiac macrophages. Moreover, fluorescence-activated cell sorting analysis (FACS) revealed that cardiac reprogramming reduced the number of chemokine receptor type 2 (CCR2)-positive inflammatory macrophages in the non-infarct areas in chronic MI, thereby restoring myocardial remodeling. Thus, cardiac reprogramming reduced the number of inflammatory macrophages to exacerbate cardiac function after MI.


Patient-Reported Outcomes Measurement Information System Correlates With Modified Harris Hip Score in Total Hip Arthroplasty.
The modified Harris Hip Score (mHHS) is a validated and disease-specific instrument commonly used to assess outcomes in total hip arthroplasty (THA). The Patient-Reported Outcomes Measurement Information System (PROMIS) is a validated, computer adaptive testing (CAT)-based global health assessment tool. The authors' goal was to examine the correlation between PROMIS Pain Interference and Physical Function CATs and the mHHS in patients undergoing primary THA. All THAs were performed by 1 of 2 fellowship-trained dedicated total joint surgeons at 1 academic institution. Patients completed PROMIS and mHHS assessments preoperatively and at 3, 6, 12, and 52 weeks postoperatively. Descriptive statistics and Pearson correlation values were determined. A total of 48 patients were prospectively enrolled in the study. Preoperatively, mean total PROMIS score (Pain Interference and Physical Function) was 74.2 and mHHS was 50.8. Preoperatively, mean total PROMIS score showed a moderate correlation (r=0.56; P<.0001) with total mHHS. Postoperatively, mean total PROMIS score at 3, 6, 12, and 52 weeks was 82.4, 93.4, 100, and 100, respectively (all P<.01 vs baseline), and mHHS was 68.2, 81.1, 85.9, and 88.6, respectively (all P<.01). At 3, 6, 12, and 52 weeks postoperatively, a strong and consistent correlation was observed between the total PROMIS score and mHHS (r=0.74, 0.74, 0.73, and 0.80, respectively; all P<.0001). The PROMIS Pain Interference and Physical Function CATs accurately assessed preoperative pain and dysfunction, as well as clinical improvement following THA. Combined PROMIS Pain Interference and Physical Function is comparable to the mHHS when assessing outcome following THA for osteoarthritis. [Orthopedics. 2021;44(1):e19-e25.].


Probing dense granular materials by space-time dependent perturbations.
The manner in which signals propagate through dense granular systems in both space and time is not well understood. In order to probe this process, we carry out discrete element simulations of the system response to excitations where we control the driving frequency and wavelength independently. Fourier analysis shows that properties of the signal depend strongly on the space-time scales of the perturbation. The features of the response provide a test bed for models that predict statistical and continuum space-time properties. We illustrate this connection between microscale physics and macroscale behavior by comparing the system response to a simple elastic model with damping.


Clinical and research issues regarding chronic advanced coronary artery disease: part I: Contemporary and emerging therapies.
The following report is based on a working group meeting about advanced coronary artery disease for patients with refractory ischemia who cannot receive revascularization. The aims were to review currently available treatment strategies, define unmet clinical needs, explore clinical trial design issues, and identify promising novel therapeutic targets and approaches for patients with chronic ischemia. The Working Group brought together medical experts in the management of refractory angina with representatives from regulatory agencies, Centers for Medicare and Medicaid Services, and industry. The meeting began with presentations reviewing the limitations of the current medical therapies and revascularization strategies and focused on lessons learned from past therapeutic attempts to optimize outcomes and on what are considered to be the most promising new approaches. Perspectives from clinical experts and from regulatory agencies were juxtaposed against needs and concerns of industry regarding development of new therapeutic strategies. This report presents the considerations and conclusions of the meeting on December 4-5, 2006. This document has been developed as a 2-part article, with contemporary and emerging therapies for advanced coronary artery disease reviewed first. Trial design, end points, and regulatory issues will be discussed in the second part of the article.


Surrogate endpoints and renal protection: focus on microalbuminuria.
End-stage renal disease (ESRD) is a major public health problem worldwide. The past decade has witnessed increased interest and investigative attention on therapeutic manoeuvres to retard the progression of renal disease. Reduction of albuminuria is regarded as a surrogate endpoint, because this phenomenon seems to be associated with preservation of GFR (principal endpoint) in diabetic nephropathy. Studies from our laboratory suggest that reduction in albuminuria during ACE inhibition constitutes a predictor of an attenuated rate of decline in GFR in early diabetic nephropathy. Because the rate of decline in kidney function is used to assess prognosis and the efficacy of therapy on progression of renal disease, a valid method for the determination of GFR is essential. During the last two decades, several radioactive and non-radioactive filtration markers have been validated and found to be both accurate and precise compared to inulin. Plasma clearance of such filtration markers is frequently applied in clinical trials in diabetic nephropathy because patients suffering from this complication frequently also suffer from diabetic cystopathy, precluding measurements of urine volume. Measurements of serum creatinine, creatinine clearance, calculated creatinine clearance by use of the Cockroft-Gault formula must all be regarded as surrogate measurements for glomerular filtration rate. Consequently, care must be taken to select appropriate filtration markers in order to rigorously evaluate therapeutic trials in progressive renal disease.


The estimation of a preference-based measure of health from the SF-12.
The SF-12 is a multidimensional generic measure of health-related quality of life. It has become widely used in clinical trials and routine outcome assessment because of its brevity and psychometric performance, but it cannot be used in economic evaluation in its current form. We sought to derive a preference-based measure of health from the SF-12 for use in economic evaluation and to compare it with the original SF-36 preference-based index. The SF-12 was revised into a 6-dimensional health state classification (SF-6D [SF-12]) based on an item selection process designed to ensure the minimum loss of descriptive information. A sample of 241 states defined by the SF-6D (of 7500) have been valued by a representative sample of 611 members of the UK general population using the standard gamble (SG) technique. Models are estimated of the relationship between the SF-6D (SF-12) and SG values and evaluated in terms of their coefficients, overall fit, and the ability to predict SG values for all health states. The models have produced significant coefficients for levels of the SF-6D (SF-12), which are robust across model specification. The coefficients are similar to those of the SF-36 version and achieve similar levels of fit. There are concerns with some inconsistent estimates and these have been merged to produce the final recommended model. As for the SF-36 model, there is evidence of over prediction of the value of the poorest health states. The SF-12 index provides a useful tool for researchers and policy makers wishing to assess the cost-effectiveness of interventions.


Transcriptional profiling reveals a role for RORalpha in regulating gene expression in obesity-associated inflammation and hepatic steatosis.
Retinoid-related orphan receptor (ROR)α4 is the major RORα isoform expressed in adipose tissues and liver. In this study we demonstrate that RORα-deficient staggerer mice (RORα(sg/sg)) fed with a high-fat diet (HFD) exhibited reduced adiposity and hepatic triglyceride levels compared with wild-type (WT) littermates and were resistant to the development of hepatic steatosis, adipose-associated inflammation, and insulin resistance. Gene expression profiling showed that many genes involved in triglyceride synthesis and storage, including Cidec, Cidea, and Mogat1, were expressed at much lower levels in liver of RORα(sg/sg) mice. In contrast, overexpression of RORα in mouse hepatoma Hepa1-6 cells significantly increased the expression of genes that were repressed in RORα(sg/sg) liver, including Sult1b1, Adfp, Cidea, and ApoA4. ChIP and promoter analysis suggested that several of these genes were regulated directly by RORα. In addition to reduced lipid accumulation, inflammation was greatly diminished in white adipose tissue (WAT) of RORα(sg/sg) mice fed with an HFD. The infiltration of macrophages and the expression of many immune response and proinflammatory genes, including those encoding various chemo/cytokines, Toll-like receptors, and TNF signaling proteins, were significantly reduced in RORα(sg/sg) WAT. Moreover, RORα(sg/sg) mice fed with an HFD were protected from the development of insulin resistance. RORα(sg/sg) mice consumed more oxygen and produced more carbon dioxide, suggesting increased energy expenditure in this genotype. Our study indicates that RORα plays a critical role in the regulation of several aspects of metabolic syndrome. Therefore, RORα may provide a novel therapeutic target in the management of obesity and associated metabolic diseases.


Spontaneous Seed Formation during Electrodeposition Drives Epitaxial Growth of Metastable Bismuth Selenide Microcrystals.
Materials with metastable phases can exhibit vastly different properties from their thermodynamically favored counterparts. Methods to synthesize metastable phases without the need for high-temperature or high-pressure conditions would facilitate their widespread use. We report on the electrochemical growth of microcrystals of bismuth selenide, Bi2Se3, in the metastable orthorhombic phase at room temperature in aqueous solution. Rather than direct epitaxy with the growth substrate, the spontaneous formation of a seed layer containing nanocrystals of cubic BiSe enforces the metastable phase. We first used single-crystal silicon substrates with a range of resistivities and different orientations to identify the conditions needed to produce the metastable phase. When the applied potential during electrochemical growth is positive of the reduction potential of Bi3+, an initial, Bi-rich seed layer forms. Electron microscopy imaging and diffraction reveal that the seed layer consists of nanocrystals of cubic BiSe embedded within an amorphous matrix of Bi and Se. Using density functional theory calculations, we show that epitaxial matching between cubic BiSe and orthorhombic Bi2Se3 can help stabilize the metastable orthorhombic phase over the thermodynamically stable rhombohedral phase. The spontaneous formation of the seed layer enables us to grow orthorhombic Bi2Se3 on a variety of substrates including single-crystal silicon with different orientations, polycrystalline fluorine-doped tin oxide, and polycrystalline gold. The ability to stabilize the metastable phase through room-temperature electrodeposition in aqueous solution without requiring a single-crystal substrate broadens the range of applications for this semiconductor in optoelectronic and electrochemical devices.


External quality assessment of BRAF molecular analysis in melanoma.
The availability of BRAF inhibitors has given metastatic melanoma patients an effective new treatment choice and molecular testing to determine the presence or absence of a BRAF codon 600 mutation is pivotal in the clinical management of these patients. This molecular test must be performed accurately and appropriately to ensure that the patient receives the most suitable treatment in a timely manner. Laboratories have introduced such testing; however, some experience low sample throughput making it critical that an external quality assurance programme is available to help promote a high standard of testing, reporting and provide an educational aspect for BRAF molecular testing. Laboratories took part in three rounds of external quality assessment (EQA) during a 12-month period giving participants a measure of the accuracy of genotyping, clinical interpretation of the result and experience in testing a range of different samples. Formalin fixed paraffin embedded tissue sections from malignant melanoma patients were distributed to participants for BRAF molecular testing. The standard of testing was generally high but distribution of a mutation other than the most common, p.(Val600Glu), highlighted concerns with detection or reporting of the presence of rarer mutations. The main issues raised in the interpretation of the results were the importance of clear unambiguous interpretation of the result tailored to the patient and the understanding that the treatment is different from that given to other stratified medicine programmes. The variability in reporting and wide range of methodologies used indicate a continuing need for EQA in this field.


[The concept of working memory in schizophrenia: current evidence and future perspectives].
Working memory (WM) deficits are core symptoms of cognitive impairment in schizophrenia. The psychological concept of WM offers a theoretical and empirical framework for the investigation of perceptual and attentional dysfunction as well as of deficits of higher cognitive functions in schizophrenia. From a theoretical perspective, schizophrenic psychopathology and cognitive dysfunction can be both integrated and experimentally validated within the concept of WM-impairment. In the last years investigation of WM has been a major issue in neuropsychiatric research, not least because of the association of prefrontal cortex and WM-function. The advent of functional imaging techniques, e. g. functional magnetic resonance imaging (fmri) has additionally contributed in generating neurobiological WM-models. Regarding the neuronal basis of WM-deficits in schizophrenia, prefrontal cortex dysfunction has been of major interest. This review provides a short overview on the concept of WM and its relevance for cognitive psychology, discussing both recent behavioral and functional neuroimaging evidence on WM-dysfunction in schizophrenia.


Affective mental contrasting to enhance physical activity: A randomized controlled trial.
Transition to university is a vulnerable period for discontinuing regular physical activity that can have implications for students' physical and psychological health. Accordingly, it is imperative to find and implement cost and time-effective interventions to mitigate the consequences of this transition. Intervention research has shown mental contrasting is an effective means of promoting various health-enhancing behaviors including physical activity. However, the efficacy of this intervention approach may be bolstered by targeting affective judgments (e.g., enjoyable-unenjoyable), which exert greater influence on physical activity behaviors compared to health-related instrumental judgments (e.g., useful-useless). The purpose of this study was to compare the relative efficacy of a mental contrasting intervention that targeted affective judgments associated with physical activity, in relation to a mental contrasting intervention that targeted instrumental judgments, and a "standard" mental contrasting intervention (with no modifications). Using a 3-arm parallel randomized controlled trial design (ClinicalTrials.gov Number NCT02615821), 110 insufficiently active, female, university students were randomly assigned to an affective, instrumental, or standard mental contrasting intervention following simple randomization procedures. Assessments were conducted at baseline and 4 weeks postintervention. Participants in the affective mental contrasting condition displayed higher levels of moderate-to-vigorous physical activity than those in the instrumental or standard comparison conditions, F(2, 90) = 3.14, p < .05, ηp2 = 0.065. Overall, affective mental contrasting has the potential to represent a low-cost and time-efficient intervention that may help insufficiently active, female students increase activity or attenuate declining levels of moderate-to-vigorous physical activity that occurs during university. (PsycINFO Database Record


Playing a part in research? University students' attitudes to direct-to-consumer genomics.
This study examined the attitudes of 1,146 Swiss University students to direct-to-consumer (DTC) genomic testing and to genomic research participation. Data were collected through a self-completion online questionnaire by students from 2 higher education institutions in Zurich, Switzerland. The survey aimed to capture motivation for undergoing or refraining from genomic testing, reactions to mock genetic risk results, and views about contributing data to scientific research. Descriptive and inferential statistics were used for the analysis. A total of 1.5% of the students had undergone testing. Most respondents were studying natural sciences and were interested in undergoing DTC genomic testing. The main motive was to contribute their data to scientific research, followed closely by their interest to find out disease risks and personal traits. Overall, 41% of the respondents were not interested in DTC tests. The primary reasons were concerns about receiving potentially worrying results. There was a significant correlation between studying natural sciences, as opposed to the humanities, and interest in undergoing testing. Male respondents were more interested in testing compared to females. There was a strong interest in genetic research participation and notably limited privacy concerns. Although 59% of the respondents were interested in DTC genomic testing, they were not likely to be affected by them or act upon them. This raises questions about concerns relating to potential risks of DTC genomics users and users' understanding of genetic information including their awareness of privacy risks. Furthermore, the strong interest in genetic research participation signals an underexplored personal utility of genomic testing which needs to be both better understood and better harnessed.


Eye Tracking for Deep Learning Segmentation Using Convolutional Neural Networks.
Deep learning with convolutional neural networks (CNNs) has experienced tremendous growth in multiple healthcare applications and has been shown to have high accuracy in semantic segmentation of medical (e.g., radiology and pathology) images. However, a key barrier in the required training of CNNs is obtaining large-scale and precisely annotated imaging data. We sought to address the lack of annotated data with eye tracking technology. As a proof of principle, our hypothesis was that segmentation masks generated with the help of eye tracking (ET) would be very similar to those rendered by hand annotation (HA). Additionally, our goal was to show that a CNN trained on ET masks would be equivalent to one trained on HA masks, the latter being the current standard approach. Step 1: Screen captures of 19 publicly available radiologic images of assorted structures within various modalities were analyzed. ET and HA masks for all regions of interest (ROIs) were generated from these image datasets. Step 2: Utilizing a similar approach, ET and HA masks for 356 publicly available T1-weighted postcontrast meningioma images were generated. Three hundred six of these image + mask pairs were used to train a CNN with U-net-based architecture. The remaining 50 images were used as the independent test set. Step 1: ET and HA masks for the nonneurological images had an average Dice similarity coefficient (DSC) of 0.86 between each other. Step 2: Meningioma ET and HA masks had an average DSC of 0.85 between each other. After separate training using both approaches, the ET approach performed virtually identically to HA on the test set of 50 images. The former had an area under the curve (AUC) of 0.88, while the latter had AUC of 0.87. ET and HA predictions had trimmed mean DSCs compared to the original HA maps of 0.73 and 0.74, respectively. These trimmed DSCs between ET and HA were found to be statistically equivalent with a p value of 0.015. We have demonstrated that ET can create segmentation masks suitable for deep learning semantic segmentation. Future work will integrate ET to produce masks in a faster, more natural manner that distracts less from typical radiology clinical workflow.


A simplified and accurate method for the analysis of urinary metabolites of testosterone-related steroids using gas chromatography/combustion/isotope ratio mass spectrometry.
The analysis of urinary metabolites of testosterone-related steroids through the measurement of their carbon isotopic signature (δ(13) C) by gas chromatography/combustion/mass spectrometry (GC/C/IRMS) is a confirmation method employed in doping control analyses. Stringent analytical conditions are essential to an accurate and precise analysis as well as the proper selection of the metabolites, which forms the basis of the refined method presented in this paper. In a simplified approach, following enzymatic hydrolysis and extraction from a relatively low volume of urine sample, a one-step high-performance liquid chromatography (HPLC) purification was developed for seven diagnostic urinary metabolites (TS) including testosterone itself, dehydroepiandrosterone, 5α- and 5β-androstanediol, epitestosterone, androsterone, etiocholanolone and two endogenous reference compounds (ERC), 5β-pregnanediol and 5α-androst-16-en-3β-ol. These steroids were pooled in three fractions and analyzed as such. With regards to the GC/C/IRMS analysis, a multi-level isotopic calibration using the 'identical treatment' principle was created. The proposed isotopic calibration yielded results for purified reference steroids with a precision ≤0.15 and accuracy of ≤0.30 ‰ (between-assay, n = 26). Compared to other common endogenous reference compounds, those selected in this study had δ(13) C values close to the target metabolites which, along with the proposed isotopic calibration, produced narrow reference intervals within ± 3‰ for most diagnostic TS-ERC pairs, in compliance with the requirements of the World Anti-Doping Agency. These carefully controlled analytical conditions are compatible with routine operations, affording accurate and precise results for the more diagnostically relevant metabolites such as testosterone itself and the 5α- and 5β-androstanediols. The values of the TS-ERC pairs measured in reference populations are described and the results from the routine testing of several hundreds of athletes' samples are discussed. Robust, this technique permitted the detection of adverse findings that would have been missed had these low level metabolites not been analyzed.


New perspectives on difficult-to-treat tuberculosis based on old therapeutic approaches.
Tuberculosis (TB) is an important clinical and public health issue worldwide. Despite improved treatment success rates following the introduction of antibiotics in daily clinical practice, the expected decline in incidence has been hampered by HIV epidemics and multi- and extensively drug-resistant TB. During the pre-antibiotic era, TB therapies were mainly based on improving hygiene conditions, strengthening the immune system, and targeting the rest of the affected lungs with invasive techniques. Detailed knowledge of old non-pharmacological therapies might support physicians and researchers in the identification of new solutions for difficult-to-treat patients. We performed a narrative literature review on the main old therapeutic options prescribed for patients with TB. The main recommendations and contraindications of sanatorium therapies (i.e., bed rest, fresh air, sunlight) and pulmonary collapse techniques are reviewed, evaluating their physiological basis and their impact on patient outcomes. We report studies describing new interventional pulmonary and surgical techniques and assess new perspectives based on old medical and surgical treatments, whose potential implementation could help complicated patients.


Understanding subtypes of inner-city drug users with a latent class approach.
We empirically identified subtypes of inner-city users of heroin and cocaine based on type of drug used and route of administration. The sample was recruited from the communities in Baltimore, MD (SHIELD study) and consisted of 1061 participants who used heroin and or cocaine in the past 6 months on a weekly basis or more. Latent class analysis (LCA) was used to identify subtypes of drug users based on type of drug and route of administration. Logistic regression was used to compare the subtypes on depressive symptoms, injection risk and drug network compositions. Inner-city drug users were classified into five subtypes: three subtypes of injection drug users (IDUs) [heroin injecting (n=134; 13%), polydrug and polyroute (n=88, 8%), and heroin and cocaine injecting (n=404, 38%)], and two subtypes with low proportions of IDUs (LIDUs) [heroin snorting (n=275, 26%) and crack smoking (n=160; 14%)]. The polydrug and polyroute subtype had the highest depressive symptoms risk among all subtypes. Injection risk was lowest in the heroin injecting subtype and significantly differed from heroin and cocaine injecting subtype. The IDU subtypes also varied in the drug network compositions. The LIDU subtypes had similar depressive symptoms risk but vastly differed in the drug network compositions. Subgroups of inner-city cocaine and heroin users based on type and route of administration differed in their depressive symptoms, injection risk and drug network compositions. Future studies should longitudinally examine factors associated with transitioning across these subtypes to better inform prevention and treatment efforts.


Mechanism of catch force: tethering of thick and thin filaments by twitchin.
Catch is a mechanical state occurring in some invertebrate smooth muscles characterized by high force maintenance and resistance to stretch during extremely slow relaxation. During catch, intracellular calcium is near basal concentration and myosin crossbridge cyctng rate is extremely slow. Catch force is relaxed by a protein kinase A-mediated phosphorylation of sites near the N- and C- temini of the minititin twitchin (approximately 526 kDa). Some catch force maintenance car also occur together with cycling myosin crossbridges at submaximal calcium concentrations, but not when the muscle is maximally activated. Additionally, the link responsible for catch can adjust during shortening of submaximally activated muscles and maintain catch force at the new shorter length. Twitchin binds to both thick and thin filaments, and the thin filament binding shown by both the N- and Cterminal portions of twitchin is decreased by phosphorylation of the sites that regulate catch. The data suggest that the twitchin molecule itself is the catch force beanng tether between thick and thin filaments. We present a model for the regulation of catch in which the twitchin tether can be displaced from thin filaments by both (a) the phosphorylation of twitchin and (b) the attachment of high force myosin crossbridges.


[Characteristics of nasopharyngeal carriage of Streptococcus pneumoniae in children during acute respiratory disease].
Streptococcus pneumoniae is an important cause of pediatric morbidity and its main reservoir is the nasopharynx, from which it can disseminate and cause invasive disease. From November 1997 through March 1998, nasopharyngeal carriage of S. pneumoniae was evaluated in 250 children under the age of 36 months: 123 Jews and 127 Bedouins with acute respiratory disease and in 980 healthy control children (852 Jews and 128 Bedouins). Carriage rate was higher among sick children. Among Jewish children it was 57% and 35% of sick and healthy children respectively (p < 0.01), and among Bedouin children it figured as 80% and 67% respectively (p = 0.01). The difference in carriage rate was most prominent in infants under the age of 5 months: among Jewish children it was 60% and 27% of sick and healthy children respectively (p < 0.001) and among Bedouins it was 82% and 65% respectively (p = 0.05). Higher carriage rate of penicillin resistant pneumococci (PRP) was also detected in sick children, with no relation to antibiotic treatment in the month prior to sampling. In Jewish children PRP was detected in 12%, 28% (p < 0.001) and 36% (p < 0.001) of healthy children, sick children with previous antibiotic treatment and sick children with no treatment, respectively. The seroypes included in the newly developed 7-valent conjugate vaccine: 4, 6B, 9V, 14, 18C, 19F, 23F, that are highly pathogenic and often antibiotic resistant contributed 74% of isolates in sick Jewish children who had previous antibiotic treatment and 39% of isolates in healthy children (p < 0.001). In Bedouin children vaccine types carriers rate among the sick children was not higher than in healthy children. Acute respiratory disease increases the risk of pneumococcal carriage in general and carriage of resistant pneumococci in particular. Previous antibiotic treatment increases the risk of carring one of the pathogenic serotypes included in the 7-valent vaccine. The impact of disease is most prominent in infants under 5 months, since they are usually less exposed to S. pneumoniae carriers than older children. Since the increase in carriage rate during illness is mostly due to the serotypes included in the newly developed conjugate vaccine, future immunization programme may decrease not only morbidity rate but also nasopharyngeal carriage rate of pneumococci in general and of antibiotic-resistant pneumococci in particular.


Functional analysis of the domain structure of tumor necrosis factor-alpha converting enzyme.
Many membrane-bound proteins, including cytokines, receptors, and growth factors, are proteolytically cleaved to release a soluble form of their extracellular domain. The tumor necrosis factor (TNF)-alpha converting enzyme (TACE/ADAM-17) is a transmembrane metalloproteinase responsible for the proteolytic release or "shedding" of several cell-surface proteins, including TNF and p75 TNFR. We established a TACE-reconstitution system using TACE-deficient cells co-transfected with TACE and substrate cDNAs to study TACE function and regulation. Using the TACE-reconstitution system, we identified two additional substrates of TACE, interleukin (IL)-1R-II and p55 TNFR. Using truncations and chimeric constructs of TACE and another ADAM family member, ADAM-10, we studied the function of the different domains of TACE in three shedding activities. We found that TACE must be expressed with its membrane-anchoring domain for phorbol ester-stimulated shedding of TNF, p75 TNFR, and IL-1R-II, but that the cytoplasmic domain is not required for the shedding of these substrates. The catalytic domain of ADAM-10 could not be functionally substituted for that of TACE. IL-1R-II shedding required the cysteine-rich domain of TACE as well as the catalytic domain, whereas TNF and p75 TNFR shedding required only the tethered TACE catalytic domain.