Small Molecule Intervention in a Protein Kinase C-Gli Transcription Factor Axis.
Aberrations in the Hedgehog (Hh) signaling pathway are responsible for a broad range of human cancers, yet only a subset rely on the activity of the clinical target, Smoothened (Smo). Emerging cases of cancers that are insensitive to Smo-targeting drugs demand new therapeutic targets and agents for inhibition. As such, we sought to pursue a recently discovered connection between the Hedgehog pathway transcription factors, the glioma-associated oncogene homologues (Glis), and protein kinase C (PKC) isozymes. Here, we report our assessment of a structurally diverse library of PKC effectors for their influence on Gli function. Using cell lines that employ distinct mechanisms of Gli activation up- and downstream of Smo, we identify a PKC effector that acts as a nanomolar Gli antagonist downstream of Smo through a mitogen-activated protein kinase kinase (MEK)-independent mechanism. This agent provides a unique tool to illuminate crosstalk between PKC isozymes and Hh signaling and new opportunities for therapeutic intervention in Hh pathway-dependent cancers.


Anesthesia and neurotoxicity study design, execution, and reporting in the nonhuman primate: A systematic review.
Concern for a role of anesthesia in neurotoxicity in children originated from neonatal rodent and nonhuman primate (NHP) models, yet prospective clinical studies have largely not supported this concern. The goal of this study was to conduct an objective assessment of published NHP study rigor in design, execution, and reporting. A MEDLINE search from 2005 to December 2021 was performed. Inclusion criteria included full-length original studies published in English under peer-reviewed journals. We documented experimental parameters on anesthetic dosing, monitoring, vitals, and experimental outcomes. Twenty-three manuscripts were included. Critical issues identified in study design included: lack of blinding in data acquisition (57%) and analysis (100%), supratherapeutic (4-12 fold) maintenance dosing in 22% of studies, lack of sample size justification (91%) resulting in a mean (SD) sample size of 6 (3) animals per group. Critical items identified in the conduct and reporting of studies included: documentation of anesthesia provider (0%), electrocardiogram monitoring (35%), arterial monitoring (4%), spontaneous ventilation employed (35%), failed intubations resulting in comingling ventilated and unventilated animals in data analysis, inaccurate reporting of failed intubation, and only 50% reporting on survival. Inconsistencies were noted in drug-related induction of neuroapoptosis and region of occurrence. Further, 67%-100% of behavior outcomes were not significantly different from controls. Important deficits in study design, execution, and reporting were identified in neonatal NHP studies. These results raise concern for the validity and reliability of these studies and may explain in part the divergence from results obtained in human neonates.


Sex differences in relations of muscle power, lung function, and reaction time in athletes.
In an earlier study, relations of nonverbal abilities with several bodily measures such as height, weight, and lung capacity were observed. The present aim was estimation of associations of muscle power and lung function with simple eye-hand reaction time. Sex differences for muscle power were significant even with the covariates of height, weight, and age included; however, these disappeared for lung functions (forced vital capacity and peak expiratory flow) and reaction time. The effects of leg power, forced vital capacity, and peak expiratory flow on the left eye-hand reaction time were significant after adjustment for height, weight, right- and left-hand powers and age. The positive effect of exercise may be especially associated with the right brain activity or left hand speed.


Pediatric Sarcoma Data Forms a Unique Cluster Measured via the Earth Mover's Distance.
In this note, we combined pediatric sarcoma data from Columbia University with adult sarcoma data collected from TCGA, in order to see if one can automatically discern a unique pediatric cluster in the combined data set. Using a novel clustering pipeline based on optimal transport theory, this turned out to be the case. The overall methodology may find uses for the classification of data from other biological networking problems.


Characterizing the space of interatomic distance distribution functions consistent with solution scattering data.
Scattering of neutrons and X-rays from molecules in solution offers alternative approaches to the study of a wide range of macromolecular structures in their solution state without crystallization. We study one part of the problem of elucidating three-dimensional structure from solution scattering data, determining the distribution of interatomic distances, P(r), where r is the distance between two atoms in the protein molecule. This problem is known to be ill-conditioned: for a single observed diffraction pattern, there may be many consistent distance distribution functions, and there is a risk of overfitting the observed scattering data. We propose a new approach to avoiding this problem: accepting the validity of multiple alternative P(r) curves rather than seeking a single "best." We place linear constraints to ensure that a computed P(r) is consistent with the experimental data. The constraints enforce smoothness in the P(r) curve, ensure that the P(r) curve is a probability distribution, and allow for experimental error. We use these constraints to precisely describe the space of all consistent P(r) curves as a polytope of histogram values or Fourier coefficients. We develop a linear programming approach to sampling the space of consistent, realistic P(r) curves. On both experimental and simulated scattering data, our approach efficiently generates ensembles of such curves that display substantial diversity.


Keratocyte survival following a controlled-rate freeze.
One of the major drawbacks of cryorefractive surgery is the delay in the postoperative recovery of corneal clarity. The authors have hypothesized that this delay is due to the death of graft keratocytes caused by the freezing process, and the need for subsequent repopulation by host keratocytes to restore corneal clarity. By using a slow, controlled-rate freeze on a modified Barraquer cryolathe, the authors are able to maintain keratocyte viability in the graft corneal tissue, as assessed by standard histologic methods and by collagenase digestion. Maintenance of keratocyte viability results in the rapid return of corneal clarity following lamellar keratoplasty in the rabbit, with few of the histopathologic changes seen following standard rapid freezing of corneal tissue.


Numeric definition of the clinical performance of the nested reverse transcription-PCR for detection of hematogenous epithelial cells and correction for specific mRNA of non-target cell origin as evaluated for prostate cancer cells.
Inappropriate quality management of reverse transcription-PCR (RT-PCR) assays for the detection of blood-borne prostate cancer (PCa) cells hampers clinical conclusions. Improvement of the RT-PCR methodology for prostate-specific antigen (PSA) mRNA should focus on an appropriate numeric definition of the performance of the assay and correction for PSA mRNA that is not associated with PCa cells. Repeated (RT-)PCR tests for PSA mRNA in single blood specimens from PCa patients and PCa-free controls, performed by four international institutions, showed a large percentage (approximately equal to 50%) of divergent test results. The best estimates of the mean, lambda (SD), of the expected Poisson frequency distributions of the number of positive tests among five replicate assays of samples from PCa-free individuals were 1.0 (0.2) for 2 x 35 PCR cycles and 0.2 (0.1) for 2 x 25 PCR cycles. Assessment of the numeric value of the mean can be considered as a new indicator of the performance of a RT-PCR assay for PSA mRNA under clinical conditions. Moreover, it determines the required number of positive test repetitions to differentiate between true and false positives for circulating prostate cells. At a predefined diagnostic specificity of > or = 98%, repeated PCRs with lambda of either 1.0 or 0.2 require, respectively, more than three or more than one positive tests to support the conclusion that PSA mRNA-containing cells are present. Repeated nested PCR tests for PSA and appropriate handling of the data allow numeric quantification of the performance of the assay and differentiation between analytical false and true positives at a predefined accuracy. This new approach may contribute to introduction of PSA RT-PCR assays in clinical practice.


Effectiveness and cost-effectiveness of community singing on mental health-related quality of life of older people: randomised controlled trial.
As the population ages, older people account for a greater proportion of the health and social care budget. Whereas some research has been conducted on the use of music therapy for specific clinical populations, little rigorous research has been conducted looking at the value of community singing on the mental health-related quality of life of older people. To evaluate the effectiveness and cost-effectiveness of community group singing for a population of older people in England. A pilot pragmatic individual randomised controlled trial comparing group singing with usual activities in those aged 60 years or more. A total of 258 participants were recruited across five centres in East Kent. At 6 months post-randomisation, significant differences were observed in terms of mental health-related quality of life measured using the SF12 (mean difference = 2.35; 95% CI = 0.06-4.76) in favour of group singing. In addition, the intervention was found to be marginally more cost-effective than usual activities. At 3 months, significant differences were observed for the mental health components of quality of life (mean difference = 4.77; 2.53-7.01), anxiety (mean difference = -1.78; -2.5 to -1.06) and depression (mean difference = -1.52; -2.13 to -0.92). Community group singing appears to have a significant effect on mental health-related quality of life, anxiety and depression, and it may be a useful intervention to maintain and enhance the mental health of older people.


Bioavailability and biodegradation of prosulfocarb in soil.
Active microbial degraders of the herbicide prosulfocarb (PSC) were isolated to evaluate their performance in soil with a view to their use for bioremediation. The isolated cultures (a microbial consortium and a Pseudomonas sp. strain) were active when tested in mineral medium with PSC as the only carbon source, but had an adverse effect on the soil indigenous microflora. Biodegradation in the inoculated soils was thus lower than in the uninoculated soil when only the indigenous microflora was present. Further tests showed that the strong affinity of PSC for soil organic matter affected its bioavailability and hence its biodegradation by the inocula. Bioremediation of PSC contaminated soils could thus be undertaken by biostimulation of indigenous microflora.


Functional characteristics of connective tissue growth factor on vitreoretinal cells.
Connective tissue growth factor (CTGF) level is elevated in eyes with proliferative vitreoretinal diseases, such as proliferative diabetic retinopathy and proliferative vitreoretinopathy (PVR), as we previously reported, but its functional characteristics on vitreoretinal cells are yet to be clarified. In this study, we demonstrated a growth-promoting effect of CTGF on cultured hyalocytes and bovine retinal pigment epithelial cells (BRPEs) with the induction of p44/p42 mitogen-activated protein kinase phosphorylation and [(3)H]thymidine incorporation. CTGF also stimulated the synthesis of fibronectin by hyalocytes and BRPEs without significant effect on collagen gel contraction by these cells. On the other hand, CTGF had no direct effects on the proliferation, migration, or in vitro tube formation by vascular endothelial cells. Nevertheless, CTGF promoted vascular endothelial growth factor (VEGF) gene expression by hyalocytes and BRPEs. Although the concentrations of both CTGF and VEGF in the human vitreous samples with proliferative vitreoretinal diseases were elevated, there was no significant correlation between these concentrations. These findings indicate that CTGF appears to be involved in the formation of proliferative membranes without direct regulation of their cicatricial contraction in the pathogenesis of proliferative vitreoretinal diseases. Whereas CTGF might have no direct effects or minimal effects, if any, on retinal neovascularization, it is possible that CTGF has indirect effects by modulating the expression of VEGF.


[Inherited thrombocytopenias].
Secondary causes of thrombocytopenia as immunologic thrombopenia purpura, or ITP, are far more common than inherited causes, which even as a group, are rare. Nevertheless, diagnosis is important and progress made in uncovering the molecular basis of these disorders has contributed greatly to our knowledge of these diseases. Inherited thrombocytopenias are a heterogeneous group of disorders. Different criteria have been suggested to classify the forms, such as the inheritance mechanism and the platelet volume as well as the associated platelet dysfunctions or clinical abnormality. This paper describes the clinical and biological data, and current knowledge of the molecular findings of inherited thrombocytopenia, allowing a diagnostic approach to these diseases.


Purification, immunological and biochemical characterization of rat 28 kDa cholecalcin (cholecalciferol-induced calcium-binding proteins). Identity between renal and cerebellar cholecalcins.
The cholecalcins are intracellular vitamin D-dependent calcium-binding proteins. High concentrations of 28 kDa cholecalcins have been found in the kidneys and cerebella of birds and mammals. However, whereas the synthesis of the renal protein is vitamin D-dependent, that of the cholecalcin in the cerebella of young growing chicks and rats is apparently not. In the present study a range of immunological, physicochemical and structural characteristics of renal and cerebellar cholecalcins isolated from a single species, the rat, has been examined to ascertain what, if any, differences there are between them, other than the vitamin D-dependence. Both proteins behaved in exactly the same way during purification and showed complete immunocross-reactivity in Ouchterlony double immunodiffusion and radioimmunoassay. No difference could be detected between them on electrophoresis under denaturing conditions or on two-dimensional isoelectric focusing/electrophoresis. Their amino acid compositions were very similar, as were their u.v.-absorption spectra and their chymotryptic and tryptic peptide maps. The N-terminal sequence was found to be Gly-Gly-Val-Ser... for both cholecalcins. We have thus been unable to show any significant difference between the two proteins and suggest that the 28 kDa cholecalcins of the rat cerebellum and kidney are extremely similar, if not identical.


WEGO: a web tool for plotting GO annotations.
Unified, structured vocabularies and classifications freely provided by the Gene Ontology (GO) Consortium are widely accepted in most of the large scale gene annotation projects. Consequently, many tools have been created for use with the GO ontologies. WEGO (Web Gene Ontology Annotation Plot) is a simple but useful tool for visualizing, comparing and plotting GO annotation results. Different from other commercial software for creating chart, WEGO is designed to deal with the directed acyclic graph structure of GO to facilitate histogram creation of GO annotation results. WEGO has been used widely in many important biological research projects, such as the rice genome project and the silkworm genome project. It has become one of the daily tools for downstream gene annotation analysis, especially when performing comparative genomics tasks. WEGO, along with the two other tools, namely External to GO Query and GO Archive Query, are freely available for all users at http://wego.genomics.org.cn. There are two available mirror sites at http://wego2.genomics.org.cn and http://wego.genomics.com.cn. Any suggestions are welcome at email@example.com.


Isolation and mutagenesis of a capsule-like complex (CLC) from Francisella tularensis, and contribution of the CLC to F. tularensis virulence in mice.
Francisella tularensis is a category-A select agent and is responsible for tularemia in humans and animals. The surface components of F. tularensis that contribute to virulence are not well characterized. An electron-dense capsule has been postulated to be present around F. tularensis based primarily on electron microscopy, but this specific antigen has not been isolated or characterized. A capsule-like complex (CLC) was effectively extracted from the cell surface of an F. tularensis live vaccine strain (LVS) lacking O-antigen with 0.5% phenol after 10 passages in defined medium broth and growth on defined medium agar for 5 days at 32°C in 7% CO₂. The large molecular size CLC was extracted by enzyme digestion, ethanol precipitation, and ultracentrifugation, and consisted of glucose, galactose, mannose, and Proteinase K-resistant protein. Quantitative reverse transcriptase PCR showed that expression of genes in a putative polysaccharide locus in the LVS genome (FTL_1432 through FTL_1421) was upregulated when CLC expression was enhanced. Open reading frames FTL_1423 and FLT_1422, which have homology to genes encoding for glycosyl transferases, were deleted by allelic exchange, and the resulting mutant after passage in broth (LVSΔ1423/1422_P10) lacked most or all of the CLC, as determined by electron microscopy, and CLC isolation and analysis. Complementation of LVSΔ1423/1422 and subsequent passage in broth restored CLC expression. LVSΔ1423/1422_P10 was attenuated in BALB/c mice inoculated intranasally (IN) and intraperitoneally with greater than 80 times and 270 times the LVS LD₅₀, respectively. Following immunization, mice challenged IN with over 700 times the LD₅₀ of LVS remained healthy and asymptomatic. Our results indicated that the CLC may be a glycoprotein, FTL_1422 and -FTL_1423 were involved in CLC biosynthesis, the CLC contributed to the virulence of F. tularensis LVS, and a CLC-deficient mutant of LVS can protect mice against challenge with the parent strain.


Role of physician extenders on the managed care team.
The use of physician extenders seems to be assured as health care reform efforts unfold at both the state and federal level. Movement to fixed budgeting and capitated reimbursement systems will encourage providers to be cost efficient whenever possible, and will accelerate the use of extenders as well as other lower cost allied professionals. As a spokesperson for Oregon's Sisters of Providence Health System said, "We have stopped investing in acute care. Our system is spending $30 million to ... establish ambulatory care settings. We need to prepare for increased numbers of patients in HMOs, more physicians who are employees, and the increased use of physician extenders." Proponents of physician extenders point to the fact that they charge less than physicians for similar services. Truly, several studies on the subject indicate that this assumption is correct. For example, a study from the Journal of Nurse Practitioners indicates that the cost of an office visit to see a nurse practitioner compared with a physician ranged from about 10% to 40% less depending on the nature of the visit (i.e., family, pediatric, gerontology, mental health, etc.). Another study has revealed that the average nurse practitioner in a family practice setting makes an annual salary of $43,600, compared to a family practice physician who makes over $110,000. While such statistics are compelling, they may not tell the entire story. For example, a study which compared APNs and primary care physicians indicated that the nurse practitioner saw fewer patients per week and spent significantly more time with each patient.(ABSTRACT TRUNCATED AT 250 WORDS)


The effectiveness of proprioceptive training for improving motor function: a systematic review.
Numerous reports advocate that training of the proprioceptive sense is a viable behavioral therapy for improving impaired motor function. However, there is little agreement of what constitutes proprioceptive training and how effective it is. We therefore conducted a comprehensive, systematic review of the available literature in order to provide clarity to the notion of training the proprioceptive system. Four major scientific databases were searched. The following criteria were subsequently applied: (1) A quantified pre- and post-treatment measure of proprioceptive function. (2) An intervention or training program believed to influence or enhance proprioceptive function. (3) Contained at least one form of treatment or outcome measure that is indicative of somatosensory function. From a total of 1284 articles, 51 studies fulfilled all criteria and were selected for further review. Overall, proprioceptive training resulted in an average improvement of 52% across all outcome measures. Applying muscle vibration above 30 Hz for longer durations (i.e., min vs. s) induced outcome improvements of up to 60%. Joint position and target reaching training consistently enhanced joint position sense (up to 109%) showing an average improvement of 48%. Cortical stroke was the most studied disease entity but no clear evidence indicated that proprioceptive training is differentially beneficial across the reported diseases. There is converging evidence that proprioceptive training can yield meaningful improvements in somatosensory and sensorimotor function. However, there is a clear need for further work. Those forms of training utilizing both passive and active movements with and without visual feedback tended to be most beneficial. There is also initial evidence suggesting that proprioceptive training induces cortical reorganization, reinforcing the notion that proprioceptive training is a viable method for improving sensorimotor function.


Spatial reasoning with external visualizations: what matters is what you see, not whether you interact.
Three experiments examined the effects of interactive visualizations and spatial abilities on a task requiring participants to infer and draw cross sections of a three-dimensional (3D) object. The experiments manipulated whether participants could interactively control a virtual 3D visualization of the object while performing the task, and compared participants who were allowed interactive control of the visualization to those who were not allowed control. In Experiment 1, interactivity produced better performance than passive viewing, but the advantage of interactivity disappeared in Experiment 2 when visual input for the two conditions in a yoked design was equalized. In Experiments 2 and 3, differences in how interactive participants manipulated the visualization were large and related to performance. In Experiment 3, non-interactive participants who watched optimal movements of the display performed as well as interactive participants who manipulated the visualization effectively and better than interactive participants who manipulated the visualization ineffectively. Spatial ability made an independent contribution to performance on the spatial reasoning task, but did not predict patterns of interactive behavior. These experiments indicate that providing participants with active control of a computer visualization does not necessarily enhance task performance, whereas seeing the most task-relevant information does, and this is true regardless of whether the task-relevant information is obtained actively or passively.


Optimized 3D Culture of Hepatic Cells for Liver Organoid Metabolic Assays.
The liver is among the principal organs for glucose homeostasis and metabolism. Studies of liver metabolism are limited by the inability to expand primary hepatocytes in vitro while maintaining their metabolic functions. Human hepatic three-dimensional (3D) organoids have been established using defined factors, yet hepatic organoids from adult donors showed impaired expansion. We examined conditions to facilitate the expansion of adult donor-derived hepatic organoids (HepAOs) and HepG2 cells in organoid cultures (HepGOs) using combinations of growth factors and small molecules. The expansion dynamics, gluconeogenic and HNF4α expression, and albumin secretion are assessed. The conditions tested allow the generation of HepAOs and HepGOs in 3D cultures. Nevertheless, gluconeogenic gene expression varies greatly between conditions. The organoid expansion rates are limited when including the TGFβ inhibitor A8301, while are relatively higher with Forskolin (FSK) and Oncostatin M (OSM). Notably, expanded HepGOs grown in the optimized condition maintain detectable gluconeogenic expression in a spatiotemporal distribution at 8 weeks. We present optimized conditions by limiting A8301 and incorporating FSK and OSM to allow the expansion of HepAOs from adult donors and HepGOs with gluconeogenic competence. These models increase the repertoire of human hepatic cellular tools available for use in liver metabolic assays.


Analysis of deoxyribonuclease activity by conjugation-free fluorescence polarisation in sub-nanolitre droplets.
We report the analysis of deoxyribonuclease (DNase) activity by conjugation-free fluorescence polarisation in a droplet-based microfluidic chip. DNase is a DNA cleaving enzyme and its activity is important in the maintenance of normal cellular functions. Alterations in DNase activity have been implicated as the cause of various cancers and autoimmune diseases. To date, various methods for the analysis of DNase activity have been reported. However, they are not cost effective due to the requirement of large sample volumes and the need for the conjugation of fluorescent dyes. In this study, we have used ethidium bromide (EtBr), a DNA intercalating reagent, as a fluorescent reporter without any prior conjugation or modification of DNA. Degradation of DNA by DNase 1 was monitored at a steady state by making changes in the fluorescence polarisation of EtBr in droplets with a volume of 330 picolitre at a 40 hertz frequency under visible light. Using this technique, we successfully determined the half-maximal inhibitory concentration (IC50) of ethylenediaminetetraacetic acid (EDTA) for the inhibition of DNase 1 activity to be 1.56 ± 0.91 mM.


Sustained JNK activation induces endothelial apoptosis: studies with colchicine and shear stress.
The disruption of microtubules by treating bovine aortic endothelial cells with 10(-7)-10(-5) M colchicine caused apoptosis, as evidenced by DNA laddering and TdT-mediated dUTP nick end labeling fluorescence staining. Colchicine treatment also induced a sustained activation of c-Jun NH(2)-terminal kinase (JNK) that lasted for >/=12 h. The blockade of JNK activity by using the negative interfering mutant JNK(K-R) markedly decreased the apoptosis induced by colchicine. Exposure of bovine aortic endothelial cells to laminar shear stress (12 dyn/cm(2)) caused a transient (<2 h) activation of JNK, and there was no induction of apoptosis. The sustained activation of JNK may play a significant role in the apoptosis induced by colchicine.


Functional categorization of unique expressed sequence tags obtained from the yeast-like growth phase of the elm pathogen Ophiostoma novo-ulmi.
The highly aggressive pathogenic fungus Ophiostoma novo-ulmi continues to be a serious threat to the American elm (Ulmus americana) in North America. Extensive studies have been conducted in North America to understand the mechanisms of virulence of this introduced pathogen and its evolving population structure, with a view to identifying potential strategies for the control of Dutch elm disease. As part of a larger study to examine the genomes of economically important Ophiostoma spp. and the genetic basis of virulence, we have constructed an expressed sequence tag (EST) library using total RNA extracted from the yeast-like growth phase of O. novo-ulmi (isolate H327). A total of 4,386 readable EST sequences were annotated by determining their closest matches to known or theoretical sequences in public databases by BLASTX analysis. Searches matched 2,093 sequences to entries found in Genbank, including 1,761 matches with known proteins and 332 matches with unknown (hypothetical/predicted) proteins. Known proteins included a collection of 880 unique transcripts which were categorized to obtain a functional profile of the transcriptome and to evaluate physiological function. These assignments yielded 20 primary functional categories (FunCat), the largest including Metabolism (FunCat 01, 20.28% of total), Sub-cellular localization (70, 10.23%), Protein synthesis (12, 10.14%), Transcription (11, 8.27%), Biogenesis of cellular components (42, 8.15%), Cellular transport, facilitation and routes (20, 6.08%), Classification unresolved (98, 5.80%), Cell rescue, defence and virulence (32, 5.31%) and the unclassified category, or known sequences of unknown metabolic function (99, 7.5%). A list of specific transcripts of interest was compiled to initiate an evaluation of their impact upon strain virulence in subsequent studies. This is the first large-scale study of the O. novo-ulmi transcriptome. The expression profile obtained from the yeast-like growth phase of this species will facilitate a multigenic approach to gene expression studies to assess their role in the determination of pathogenicity for this species. The identification and evaluation of gene targets in such studies will be a prerequisite to the development of biological control strategies for this pathogen.


Breast cancer in Brazil: epidemiology and treatment challenges.
Notwithstanding the advances in tumor research, diagnosis, and treatment, breast cancer is still a challenge worldwide. This global burden of disease has been associated with population aging and the persistence of cancer-related behaviors. The number of women diagnosed with breast cancer has been estimated as increasing, especially in middle-income countries such as Brazil. Estimates from the Instituto Nacional de Câncer (INCA) point to breast cancer as the major malignant neoplasia in Brazilian women and the main cause of death from cancer in the country. This fact has been associated with increased life expectancy, urbanization, and cancer-related behaviors. Given this scenario, it is clear that there is a need for identifying and discussing which factors have substantially contributed to this growing number of cases in Brazil, including access to treatment, prevention and early diagnosis, weaknesses of the local health policy, and intrinsic genetic peculiarities of the Brazilian population. This review aims to address the role of such factors.


Inhibition of maturation of human monocyte-derived dendritic cells in a patient with Mycobacterium avium infection.
Mycobacterium avium complex is a facultative intracellular pathogen that can cause pulmonary disease in immunocompromised individuals. Dendritic cells (DCs) play a central role in protective immunity against mycobacteria. Mycobacterium avium complex infects DCs but does not impair in vitro infected monocytes differentiation into DCs. A 54-year old woman affected by chronic graft-versus-host-disease (cGVHD) was referred to our Division of Dermatology. Immature DCs were generated from her monocytes. One week later she was hospitalized due to a lung infection with Mycobacterium avium complex. Monocyte-derived DCs during Mycobacterium avium infection expressed low levels of CD1a and CD80 as determined by flow cytometry. They also expressed high levels of CD83 and CD86, and when stimulated with LPS for 24 hrs they slightly up-regulated CD83 and did not produce IL12. When monocyte-derived DCs were obtained from the patient after having recovered from the Mycobacterium avium complex infection, they expressed normal levels of CD1a and CD80 and were negative both for CD83 and for CD86. IL12 production in response to LPS was restored. Inhibition of DC maturation by the in vivo infection with Mycobacterium avium may be an immune-evasion mechanism used by the pathogen because incompletely matured DCs may not activate effector T cells efficiently in vivo.


Neon color spreading in dynamic displays: temporal factors.
When a red star is placed in the middle of an Ehrenstein figure so as to be collinear with the surrounding black rays, a reddish veil is perceived to fill the white center. This is called neon color spreading. To better understand the processes that give rise to this phenomenon, we studied the temporal properties of the effect. Specifically, we presented a "sustained" black Ehrenstein figure (rays) for 600 ms and a "transient" red star for 48 ms, or the converse pattern, at various stimulus onset asynchronies (-100-700 ms) and asked subjects to compare the strength of the neon color in the test stimulus to that of a reference pattern in which the transient star had an onset asynchrony of 300 ms. Additional exposure durations of 24 and 96 ms were used for each transient stimulus in order to study the effect of temporal integration. Simultaneity of the on- and off-transients of the star and the Ehrenstein rays were found to optimize neon color spreading, especially when both stimuli terminated together. Longer exposure durations of the transient stimulus up to 96 ms further improved the effect. Neon color spreading was much reduced when the transient stimulus was presented soon after the beginning of the sustained stimulus, with a gradual build-up towards the end. These results emphasize the importance of stimulus onset asynchrony (SOA) and stimulus termination asynchrony (STA) for the perception of neon color spreading.


[15N]-labeled pea flour protein nitrogen exhibits good ileal digestibility and postprandial retention in humans.
The aim of the present study was to evaluate postprandial absorption of pea protein as well as exogenous nitrogen retention in humans. For this purpose, after fasting overnight, seven healthy adults (4 males and 3 females) ingested [15N]-labeled pea protein (195 mmol N). Ileal effluents were collected for 8 h at 30-min intervals using a nasointestinal intubation technique. Urine and plasma samples were collected for 24 h. The [15N]-enrichment was determined in the intestinal samples, in the plasma amino acids and urea as well as in the urinary urea and ammonia fractions. The true gastroileal absorption of pea protein was 89.4 +/- 1.1%. This absorption was correlated with a significant increase (P < 0.05) in [15N]-enrichment in the plasma amino acids and in the nitrogen incorporated into the body urea pool for 1 h following pea ingestion. The enrichment remained significantly higher than the basal values in these pools 24 h after pea ingestion. The recovery of total urinary exogenous nitrogen after 22 h was 31.1 +/- 9.3 mmol N. Moreover, the kinetics of [15N]-labeled pea amino acids deamination reached a plateau of 39 mmol. Under these conditions, pea nitrogen retention represented 78% of the absorbed dietary nitrogen in healthy humans. The present results demonstrate the good true nitrogen digestibility and retention of pea protein in humans.


Frequency and distribution of Notch mutations in tumor cell lines.
Deregulated Notch signaling is linked to a variety of tumors and it is therefore important to learn more about the frequency and distribution of Notch mutations in a tumor context. In this report, we use data from the recently developed Cancer Cell Line Encyclopedia to assess the frequency and distribution of Notch mutations in a large panel of cancer cell lines in silico. Our results show that the mutation frequency of Notch receptor and ligand genes is at par with that for established oncogenes and higher than for a set of house-keeping genes. Mutations were found across all four Notch receptor genes, but with notable differences between protein domains, mutations were for example more prevalent in the regions encoding the LNR and PEST domains in the Notch intracellular domain. Furthermore, an in silico estimation of functional impact showed that deleterious mutations cluster to the ligand-binding and the intracellular domains of NOTCH1. For most cell line groups, the mutation frequency of Notch genes is higher than in associated primary tumors. Our results shed new light on the spectrum of Notch mutations after in vitro culturing of tumor cells. The higher mutation frequency in tumor cell lines indicates that Notch mutations are associated with a growth advantage in vitro, and thus may be considered to be driver mutations in a tumor cell line context.


Using nursing science to advance policy and practice in the context of social and structural determinants of health.
Social and structural determinants of health play a large role in health inequities. To highlight how nursing science can be used to advance policy and practice in the context of social and structural determinants of health. This paper reports on the author's keynote presentation from the 2022 State of The Science Conference on Social and Structural Determinants of Health presented by the Council for the Advancement of Nursing Science. Key concepts are overviewed and defined, followed by examples of two community-engaged research projects with findings that inform practice and policy. The author concludes with individual-, social- and structural-level recommendations as a clarion call for nurses to use research to eliminate health inequities and promote justice for all. What we know is, in part, only as good as what we do with that knowledge. When lives are at stake, gone are the days of knowing something and failing to act on that knowledge.


Potential role for retinoic acid receptor-gamma in the inhibition of breast cancer cells by selective retinoids and interferons.
Retinoids are known to inhibit the growth of a wide variety of cancer cells, including breast cancer cells. Advances made in recent years in the understanding of the molecular mechanisms of retinoid action have allowed the design of retinoids with selective activities. Such selective retinoids are of particular interest, because they may reduce the number of undesirable side effects observed with natural compounds. Here, we have compared the growth-inhibitory activities of natural retinoids with various selective retinoids, including anti-activator protein (AP)-1 selective compounds on estrogen receptor-positive and -negative breast cancer cell lines. In addition, we have investigated cooperativity between selective retinoids and IFNs and have begun to analyze the pathways that these two different growth inhibitors use for antagonizing breast cancer cell proliferation. We observe that several selective retinoids can inhibit breast cancer cells as efficiently as the natural compounds. Anti-AP-1-selective retinoids are as effective as retinoic acid receptor (RAR)-beta/gamma-selective compounds. This lets us conclude that retinoid-induced inhibition of breast cancer cell growth does not require retinoid receptor transactivation. Several synthetic retinoids including anti-AP-1-selective compounds show synergism with IFNs. However, true synergism between the two different types of growth regulators was seen only when both classes of molecules were used at low concentrations. RAR-beta/gamma and anti-AP-1-selective retinoids, but not RAR-alpha-selective compounds, induced increased RAR-gamma mRNA levels. Interestingly, IFNs at elevated concentrations (100 units/ml and higher) also induced increased RAR-gamma expression. Thus, when used at high concentrations, IFNs may activate growth-inhibitory pathways overlapping with those activated by retinoids. Because increased RAR-gamma expression is induced by the two different classes of breast cancer cell inhibitors, it is likely to have an important role in controlling the growth fo these cancer cells.


Clinical roundtable monograph: effective management of quality of life in metastatic breast cancer.
Quality of life is accepted as an important consideration in the management of patients with metastatic breast cancer, which remains incurable. Recent clinical trials of newer agents, such as eribulin and trastuzumab emtansine, have incorporated quality of life analyses. Quality of life is impacted by multiple patient-related, disease-related, and treatment-related factors. Therapies most beneficial for maintaining or improving quality of life include those that can effectively reduce tumor burden and tumor-related symptoms, but have toxicity profiles that are well tolerated and easily managed. Overall outcomes of patients with metastatic breast cancer improve when therapy is focused not only on the disease itself, but also on the goals of minimizing diseaserelated and treatment-related symptoms. A paradigm shift now reflected in major guidelines is the incorporation of palliative care strategies earlier in the course of metastatic disease management. The selection and sequence of treatments should be made in cooperation with the patient and after consideration of her particular priorities.


Robot-Assisted and Manual Cochlear Implantation: An Intra-Individual Study of Speech Recognition.
Cochlear implantation (CI) allows rehabilitation for patients with severe to profound hearing impairment. Although the use of a robotic assistant provides technical assistance to the surgeon, the assessment of the impact of its use on auditory outcomes remains uncertain. We aim to compare the hearing results of patients who underwent bilateral cochlear implantation; one side was performed with manual insertion and the other side with robot-assisted insertion. The electrode array intrascalar positioning and the surgery duration were also studied. This retrospective intra-individual study involved 10 patients who underwent bilateral cochlear implantation. The study included two infants and eight adults. The unique composition of this cohort enabled us to utilize each patient as their own control. Regarding speech disyllabic recognition, pure tone average, ECAP, ratio of array translocation, basilar membrane rupture, and percentage of translocated electrodes, there was no difference between manual and robot-assisted CI groups. This study is the first to compare intra-individual hearing performance after cochlear implantation, either manually or robot-assisted. The number of patients and the time delay between manual and robotic implantation may have led to a lack of power, but there was no apparent difference in hearing performance between manual and robotic implantation.


Homocysteine thiolactone and protein homocysteinylation in human endothelial cells: implications for atherosclerosis.
Editing of the nonprotein amino acid homocysteine by certain aminoacyl-tRNA synthetases results in the formation of the thioester homocysteine thiolactone. Here we show that in the presence of physiological concentrations of homocysteine, methionine, and folic acid, human umbilical vein endothelial cells efficiently convert homocysteine to thiolactone. The extent of this conversion is directly proportional to homocysteine concentration and inversely proportional to methionine concentration, suggesting involvement of methionyl-tRNA synthetase. Folic acid inhibits the synthesis of thiolactone by lowering homocysteine and increasing methionine concentrations in endothelial cells. We also show that the extent of post-translational protein homocysteinylation increases with increasing homocysteine levels but decreases with increasing folic acid and HDL levels in endothelial cell cultures. These data support a hypothesis that metabolic conversion of homocysteine to thiolactone and protein homocysteinylation by thiolactone may play a role in homocysteine-induced vascular damage.


CpG and non-CpG oligodeoxynucleotides directly costimulate mouse and human CD4+ T cells through a TLR9- and MyD88-independent mechanism.
TLR ligands are known to activate APCs, but direct T cell responsiveness to TLR ligands is controversial. Because of their clinical relevance, we performed in-depth studies of the effects of the TLR9-associated ligands, oligodeoxynucleotides (ODNs), on highly purified T lymphocytes. Both CpG and non-CpG ODNs directly costimulate mouse and human CD4(+) T cells, resulting in activation marker upregulation, cytokine secretion, elevated TCR phosphorylation, and proliferation. Surprisingly, ODN costimulation occurred independently of TLR9 and MyD88, as well as ICOS, CD28, and TRIF. TLR9-antagonist ODNs likewise promoted T cell activation, which has important implications for the study of these "inhibitory" ODNs in inflammatory diseases. Cytokine profiling revealed that ODNs promote polarization of distinct Th subsets, and that ODNs differentially affect human naive and memory T cells. Our studies reveal a striking and unexpected ability of ODNs to directly activate and polarize T cells, presenting an opportunity to enhance the paradigm for selection of therapeutic ODNs in humans.


Generation of a bank of clinical-grade, HLA-homozygous iPSC lines with high coverage of the Spanish population.
Induced pluripotent stem cell (iPSC)-derived cell therapies are an interesting new area in the field of regenerative medicine. One of the approaches to decrease the costs of iPSC-derived therapies is the use of allogenic homozygous human leukocyte antigen (HLA)-matched donors to generate iPSC lines and to build a clinical-grade iPSC bank covering a high percentage of the Spanish population. The Spanish Stem Cell Transplantation Registry was screened for cord blood units (CBUs) homozygous for the most common HLA-A, HLA-B and HLA-DRB1 haplotypes. Seven donors were selected with haplotypes covering 21.37% of the haplotypes of the Spanish population. CD34-positive hematopoietic progenitors were isolated from the mononuclear cell fraction of frozen cord blood units from each donor by density gradient centrifugation and further by immune magnetic labeling and separation using purification columns. Purified CD34 + cells were reprogrammed to iPSCs by transduction with the CTS CytoTune-iPS 2.1 Sendai Reprogramming Kit. The iPSCs generated from the 7 donors were expanded, characterized, banked and registered. Master cell banks (MCBs) and working cell banks (WCBs) from the iPSCs of each donor were produced under GMP conditions in qualified clean rooms. Here, we present the first clinical-grade, iPSC haplobank in Spain made from CD34 + cells from seven cord blood units homozygous for the most common HLA-A, HLA-B and HLA-DRB1 haplotypes within the Spanish population. We describe their generation by transduction with Sendai viral vectors and their GMP-compliant expansion and banking. These haplolines will constitute starting materials for advanced therapy medicinal product development (ATMP).


A case of profound neutropenia and agranulocytosis associated with off-label use of ceftaroline.
A case of profound neutropenia and agranulocytosis associated with the off-label use of ceftaroline is reported. A 67-year-old Caucasian man arrived at the emergency room with right shoulder pain and weakness that radiated to his right chest, back, and right arm. A review of symptoms was notable for two days of burning with urination associated with decreased urinary output and decreased appetite. Multiple tests revealed the presence of methicillin-resistant Staphylococcus aureus (MRSA) septic arthritis, which was treated with an off-label dosage of ceftaroline (600 mg intravenously every eight hours). At the start of ceftaroline therapy, the patient's baseline absolute neutrophil count (ANC) was 6640 cells/μL and decreased to 816 cells/μL by day 19, eventually falling to 0 cells/μL on day 21 of therapy. Ceftaroline was then discontinued due to the suspicion that the neutropenia was secondary to maturation arrest of the bone marrow. The patient was switched to i.v. daptomycin to finish a six-week course of antibiotics. Interventional radiology placed a drain in the patient's right shoulder during the hospital stay, with symptom improvement. His white blood cell count continued to increase after ceftaroline discontinuation, reaching 6.5×10(3) cells/μL with a differential of 56.6% segmented neutrophils and 28.4% lymphocytes after nine days off of ceftaroline. A 67-year-old man developed profound neutropenia and agranulocytosis after three weeks of high-dose ceftaroline therapy for the treatment of MRSA septic arthritis. His neutropenia resolved after ceftaroline discontinuation and treatment with an alternative antibiotic.


[Current aspects of treatment for clubfoot in infants and young children].
The congenital club-foot is one of the frequent malformations (incidence around 2%). The treatment during early childhood implies conservative measures with corrective manipulations and casting, surgical procedures with extensive soft tissue releases. The technically demanding medial release is a well established procedure; however it is often not performed because the peritalar malpositions are often underestimated in the young child. After surgical correction stabilization of the foot for prolonged time is important and follow-up is mandatory until adulthood is reached.


Engineering and the law-the wages of sin: preventing punitive damages.
The author discusses the common themes in several recent cases holding medical device manufacturers liable for punitive damages. He points out that current medical device design and manufacturing standards prevent clearly dangerous products from staying in the marketplace, creating the legal expectation that devices should be risk-free. A court that expects devices to be risk-free may misunderstand the inevitable adverse product reports that accompany rigorous product testing. Therefore, device manufacturers must anticipate and manage these misunderstandings during product development if they are to appear credible before the court. A framework for a preventive law plan to minimize the risk of punitive damage awards is presented.


Gadolinium-DTPA in MR imaging of glioblastomas and intracranial metastases.
In 14 patients with the diagnosis of glioblastoma (n = 7) or intracranial metastases (n = 7), magnetic resonance (MR) imaging was performed using a variety of spin-echo (SE) pulse sequences before and after intravenous injection of 0.1 mmol gadolinium-DTPA (Gd-DTPA) per kilogram of body weight. In 10 patients, tumor tissue could not be adequately differentiated from perifocal edema on unenhanced scans with any of the applied pulse sequences. In four cases of intracranial metastases, poor differentiation between tumor and perifocal edema was possible in T2-weighted (SE 1600/70 and SE 1600/105) unenhanced scans. After administration of Gd-DTPA, tumor tissue showed marked contrast enhancement, and tumor delineation was consistently possible on SE 800/35 images. Tumor tissue could be differentiated from perifocal edema on SE 800/70 scans. Gd-DTPA is likely to increase the potential of MR imaging and refine the evaluation of glioblastomas and intracerebral metastases.


Unresectable Pancreatic Adenocarcinoma: Eight Years Later.
Pancreatic cancer is the fourth leading cause of cancer deaths in the United States, and is considered uniformly fatal when patients present with unresectable, advanced-stage disease at the time of diagnosis. Long-term survival of patients with advanced-stage pancreatic adenocarcinoma remains rare, despite advances in adjuvant chemoradiation protocols. A 73-year-old male presented to our emergency department with abdominal pain and a history of biopsy-proven, stage III pancreatic adenocarcinoma. His initial staging CT scan and trans-duodenal ultrasound had demonstrated a stage IIa (T3, N0, Mx) lesion. On surgical exploration, he was up-staged to stage III (T4, N0, Mx), noting encasement of the superior mesenteric vessels and involvement of the portal vein. He underwent palliative choledochojejunostomy and was treated with 4 months of oxaliplatin and capecitabine, with concurrent radiation therapy (50.4 Gy), followed by 4 months of gemcitabine. After 7 months, the patient withdrew from therapy due to treatment intolerance. He then turned to self-medication with non-traditional herbal therapies. After 3 years of surveillance, he was lost to follow-up until presenting to our facility with abdominal pain 8 years after his initial diagnosis. On diagnostic CT scan during his current presentation for abdominal pain, he was found to have no evidence of pancreatic cancer. Based on our review of the literature, we present the longest known survival of a patient with surgically unresectable pancreatic adenocarcinoma. Further study of this patient's phenotypic or genotypic characteristics may provide insight into better therapeutic agents, or a predictive subset of patients who will benefit from specific chemotherapeutic options.


TGRL Lipolysis Products Induce Stress Protein ATF3 via the TGF-β Receptor Pathway in Human Aortic Endothelial Cells.
Studies have suggested a link between the transforming growth factor beta 1 (TGF-β1) signaling cascade and the stress-inducible activating transcription factor 3 (ATF3). We have demonstrated that triglyceride-rich lipoproteins (TGRL) lipolysis products activate MAP kinase stress associated JNK/c-Jun pathways resulting in up-regulation of ATF3, pro-inflammatory genes and induction of apoptosis in human aortic endothelial cells. Here we demonstrate increased release of active TGF-β at 15 min, phosphorylation of Smad2 and translocation of co-Smad4 from cytosol to nucleus after a 1.5 h treatment with lipolysis products. Activation and translocation of Smad2 and 4 was blocked by addition of SB431542 (10 μM), a specific inhibitor of TGF-β-activin receptor ALKs 4, 5, 7. Both ALK receptor inhibition and anti TGF-β1 antibody prevented lipolysis product induced up-regulation of ATF3 mRNA and protein. ALK inhibition prevented lipolysis product-induced nuclear accumulation of ATF3. ALKs 4, 5, 7 inhibition also prevented phosphorylation of c-Jun and TGRL lipolysis product-induced p53 and caspase-3 protein expression. These findings demonstrate that TGRL lipolysis products cause release of active TGF-β and lipolysis product-induced apoptosis is dependent on TGF-β signaling. Furthermore, signaling through the stress associated JNK/c-Jun pathway is dependent on TGF-β signaling suggesting that TGF-β signaling is necessary for nuclear accumulation of the ATF3/cJun transcription complex and induction of pro-inflammatory responses.


Temporal changes in activity during destruction of the thoracic ventral eclosion muscle of the tsetse fly.
The spontaneous intracellular activity of the thoracic ventral longitudinal eclosion muscle (VLEM) of Glossina is described for the period from eclosion up to a short time before the final breakdown of recorded fibres. The VLEM comprises a single motor unit with no inhibitory input. The firing frequency of the motor unit declines over 5 h after eclosion and leg release. Over a period of inactivity lasting between 19-24 h in the sample fibres, there is no loss of resting membrane potential and occasional miniature potentials. The inactivity is ended by the sudden onset of biphasic potentials very different in form to the motor potentials and having a greatly reduced amplitude. These potentials fired at 6 Hz. lasted 2-4 h and ended with a rise in frequency to 25 Hz. No further activity is recorded and the fibres are observed to lose their striations and rigor. Experiments to characterize the ionic basis of activity in the VLEM have been done on spontaneous and evoked activity. Like other insect muscles, the VLEM has a major Ca2+ potential but unlike insect skeletal muscles, it also appears to have a TTX-sensitive component. This Na+ component is revealed by pretreating the system in Na(+)-free-choline saline, or by treatment with TEA in Ca(2+)-free saline. Neither EGTA nor cobalt abolish this potential. Addition of EGTA does not inhibit nerve evoked activity suggesting that the VLEM neuromuscular junction is in some way protected. The similarity of this muscle to insect visceral muscles is discussed.


A Survey for End-Stage Ankle Arthritis Treatment: Ankle Arthrodesis Versus Ankle Arthroplasty.
With promising technological advances, ankle arthroplasty has become an alternative to arthrodesis, traditionally the gold standard, for treating end-stage ankle arthritis. We collected knowledge and perceptions on both procedures to determine the need for a patient decision aid for these patients by administering a cross-sectional survey to 103 orthopaedic surgeons. Respondents were predominantly male and 41 to 50 years old. Half of those who stated that they do not perform arthroplasty said this was because they do not have adequate training. Additionally, certain variables were associated with the surgeon's choice of intervention: patient gender, age, body mass index, postoperative activity level, employment type, perceived risk of infection, neurovascular injury or wound complication, risk of developing or pre-existing adjacent arthritis, deformity, malalignment, bone loss or abnormal bone quality, number of prior ankle operations, cause of arthritis, and desire for motion preservation. The majority agreed that they always incorporate patient preferences into their decisions and that a decision aid would be beneficial. This survey revealed that several patient characteristics are influential in the surgeon's preference for either arthroplasty or arthrodesis for end-stage ankle arthritis. Because the majority of surgeons incorporate patient preferences in their decisions and report that a decision aid would be beneficial for informed decision-making in this clinical scenario, this survey identified an unmet need supporting the development of such a tool for these patients.


Short-window spectral analysis of cortical event-related potentials by adaptive multivariate autoregressive modeling: data preprocessing, model validation, and variability assessment.
In this article we consider the application of parametric spectral analysis to multichannel event-related potentials (ERPs) during cognitive experiments. We show that with proper data preprocessing, Adaptive MultiVariate AutoRegressive (AMVAR) modeling is an effective technique for dealing with nonstationary ERP time series. We propose a bootstrap procedure to assess the variability in the estimated spectral quantities. Finally, we apply AMVAR spectral analysis to a visuomotor integration task, revealing rapidly changing cortical dynamics during different stages of task processing.


Subchronic toxicity of 2,2',3,3',4,4'-hexachlorobiphenyl in rats.
The subchronic toxicity of 2,2',3,3',4,4'-hexachlorobiphenyl (PCB 128) was investigated in rats following dietary exposure at 0, 0.05, 0.5, 5, or 50 ppm for 13 wk. The growth rate was not affected by treatment and no apparent clinical signs of toxicity were observed. There was a significant increase in liver weight in the 50 ppm females. The liver ethoxyresorufin deethylase (EROD) activity was increased by five- and fourfold in the highest dose males and females, respectively, while aminopyrine demethylase (ADPM) activity was significantly increased only in the highest dose females. Liver vitamin A was significantly reduced in the highest dose females. No other biochemical or hematological effects were observed. Treatment-related histopathological changes were seen in the thyroid and liver, and to a lesser extent in the bone marrow and thymus. Residue data showed a dose-dependent accumulation of PCB 128 in the following tissues: fat, liver, kidney, brain, spleen, and serum, with the highest concentration being found in fat followed by liver and kidney. Based on these data, the no-observable-adverse-effect level of PCB 128 was judged to be 0.5 ppm in diet or 42 micrograms/kg body weight.


Multinuclear NMR and MRI Reveal an Early Metabolic Response to mTOR Inhibition in Sarcoma.
Biomarkers predicting rapalog responses in sarcomas where PI3K and mTOR are often hyperactivated could improve the suitable recruitment of responsive patients to clinical trials. PI3K/mTOR pathway activation drives energy production by regulating anaerobic glycolysis in cancer cells, suggesting a route toward a monitoring strategy. In this study, we took a multimodality approach to evaluate the phenotypic effects and metabolic changes that occur with inhibition of the PI3K/mTOR pathway. Its central role in regulating glycolysis in human sarcomas was evaluated by short- and long-term rapamycin treatment in sarcoma cell lines. We observed an overall decrease in lactate production in vitro, followed by cell growth inhibition. In vivo, we observed a similar quantitative reduction in lactate production as monitored by hyperpolarized MRI, also followed by tumor size changes. This noninvasive imaging method could distinguish reduced cell proliferation from induction of cell death. Our results illustrate the use of hyperpolarized MRI as a sensitive technique to monitor drug-induced perturbation of the PI3K/mTOR pathway in sarcomas. Cancer Res; 77(11); 3113-20. ©2017 AACR.


New approaches to structure-based discovery of dengue protease inhibitors.
Dengue virus (DENV), a member of the family Flaviviridae, presents a tremendous threat to global health since an estimated 2.5 billion people worldwide are at risk for epidemic transmission. DENV infections are primarily restricted to sub-tropical and tropical regions; however, there is concern that the virus will spread into new regions including the United States. There are no approved antiviral drugs or vaccines to combat dengue infection, although DENV vaccines have entered Phase 3 clinical trials. Drug discovery and development efforts against DENV and other viral pathogens must overcome specificity, efficacy, safety, and resistance challenges before the shortage of licensed drugs to treat viral infections can be relieved. Current drug discovery methods are largely inefficient and thus relatively ineffective at tackling the growing threat to public health presented by emerging and remerging viral pathogens. This review discusses current and newly implemented structure-based computational efforts to discover antivirals that target the DENV NS3 protease, although it is clear that these computational tools can be applied to most disease targets.


Molecular epidemiology of vancomycin-resistant enterococci: a 2-year perspective.
To determine the molecular epidemiology of vancomycin-resistant enterococci (VRE) at our medical center in order to identify the extent of strain clonality and possible transmission patterns of this pathogen. An important facet of our infection control program includes molecular typing of all clinical and surveillance isolates of VRE to determine transmission patterns in the hospital. Molecular strain typing is performed by restriction endonuclease analysis (REA) of genomic DNA. REA patterns are visually compared to categorize VRE strains into type and subtype designations. A 588-bed, university-affiliated, tertiary-care hospital and a neighboring 155-bed rehabilitation facility. From January 1995 through December 1996, 379 VRE isolates were collected from 197 patients. Thirty-three genotypes were determined by REA typing; 15 genotypes were implicated in 29 instances of potential nosocomial transmission. Three major clusters of VRE involving patients on multiple nursing units and two adjacent hospitals were identified. The remaining instances of nosocomial transmission occurred in small patient clusters. In conclusion, the VRE epidemic at this medical center is polyclonal. VRE transmission patterns are complex, and, while large clusters do occur, the usual pattern of nosocomial acquisition of this pathogen occurs in the setting of "mini-clusters".


Effects of adding minimally refined cottonseed oil or crude glycerol to diets containing 40% corn distiller's dried grains with solubles on growth performance, carcass characteristics, and pork fat firmness of growing-finishing pigs.
Diets containing more than 20% distiller's dried grains with solubles (DDGS) reduce fat firmness in pork, but supplementation of cottonseed oil or crude glycerol may improve fat firmness. The objective of this study was to assess the effect of feeding minimally refined cottonseed oil or crude glycerol on growth performance, carcass composition, and fat quality of growing-finishing pigs. Mixed sex pigs ( = 216; 24 ± 4 kg initial BW) were blocked by BW and allotted to 1 of 3 dietary treatments: 1) a basal corn-soybean meal diet with 40% DDGS (CON), 2) CON diet plus 5% minimally refined cottonseed oil added throughout the experiment (COT), or 3) CON fed during the first 8 wk and CON + 8% crude glycerol fed during the last 6 wk of the experiment (GLY). Although diets were not isocaloric, total AA-to-ME ratios were calculated to be equal among diets. Carcass composition was estimated using real-time ultrasound 2 d before harvest. Gilts (16/treatment) closest to the mean BW of each pen were harvested (115 ± 8 kg BW), and bellies were retrieved for in-depth analysis of fat quality. Belly fat was sampled and analyzed for fatty acid composition. Overall, ADFI of pigs fed COT (2.30 kg/d) was less ( < 0.01) than that of pigs fed CON or GLY (2.47 and 2.49 kg/d, respectively). Pigs fed COT (0.93 kg/d) had greater ( < 0.01) ADG compared with pigs fed CON or GLY (0.88 and 0.87 kg/d, respectively). Greater ( < 0.01) G:F was observed for pigs fed COT (0.41) than for pigs fed CON or GLY diets (0.36 and 0.35, respectively). Final BW of pigs fed COT (124.3 kg) was greater ( < 0.01) than that of pigs fed CON or GLY (118.9 and 118.6 kg, respectively). Pigs fed COT had greater ( < 0.01) HCW (94.9 kg) compared with pigs fed CON or GLY (89.9 and 89.2 kg, respectively). No differences were observed for dressing percentage (75.7, 76.3, and 75.3%), fat-free carcass lean percentage (50.5, 49.7, and 50.0%), and belly flop angle (6.21, 8.57, and 6.06°) for CON, COT, and GLY, respectively. Pigs assigned to COT had higher ( < 0.01) melting point of belly fat compared with pigs assigned to CON or GLY (30.4 vs. 26.3 and 25.3°C, respectively). Pigs fed COT had increased ( < 0.05) SFA, PUFA, and iodine value (IV) compared with CON-fed pigs. Glycerol supplementation had no influence on SFA, MUFA, and PUFA concentrations or IV of belly, jowl, and back fat compared with CON. In conclusion, COT diets improved growth performance due to greater energy density, but carcass composition was not affected by treatments. In this experiment, feeding neither COT nor GLY improved fat firmness of pigs fed diets containing 40% DDGS.


Pressure-Regulated Excitonic Transitions in Emergent Metal Halides.
ConspectusEmergent metal halides are generating significant interest as novel optical materials, and their diverse applications have brought them to the spotlight of chemistry and material science. The optical properties of semiconducting metal halides are fundamentally dominated by excitonic transitions, which refer to the complex processes of excitonic formation, self-trapping, as well as subsequent transitions of intersystem crossing (ISC) and internal conversion (IC). In this regard, high pressure has recently opened a new research dimension to regulate excitonic transitions in metal halides via continuous structural modulations, to understand the intriguing excitonic emissions from a new perspective. In this Account, we aim to rationalize the fundamental strategy for modulating and optimizing the optical properties of metal halides based on delicate exciton regulation via high-pressure method. First, the band gaps of metal halides that are directly related to the efficiency of excitonic formation, are accurately modulated through contraction, distortion, and destruction of metal-halogen polyhedra under compression. Then, considerable enhancement of self-trapped exciton emission is demonstrated by inducing proper polyhedral distortions via high-pressure method. Furthermore, the emission energy of metal halides could also be controllably and widely tuned through pressure-modulated excitonic transitions. Upon compression on different metal halides, excitonic IC is promoted with sufficient polyhedral distortions, and different sets of ISC could also be achieved. In the end, we emphasize the significance of high-pressure investigations in uncovering the complex excitonic transitions in emergent metal halides and predicting novel metal halides with desired optical properties at ambient conditions. It is expected that these discussions could inspire researchers in different fields to perform interdisciplinary high-pressure studies on novel functional materials.


Economic hardship and sexually transmitted diseases in Haiti's rural Artibonite Valley.
A study was conducted to determine the prevalence rate and risk factors for sexually transmitted diseases (STDs) in Haiti's rural Artibonite Valley. Women attending antenatal services at Hospital Albert Schweitzer from October to December 1996 were tested for gonorrhea, chlamydia, trichomonas, syphilis, and human immunodeficiency virus (HIV). Of the 476 women tested, 121 (25.4%) had trichomonas, 11/475 (2.3%) had gonorrhea, 51/475 (10.7%) had chlamydia, 32/474 (6.8%) were seropositive for syphilis, 20/469 (4.3%) were seropositive for HIV, and 191 (40.1%) had at least one STD. Nearly 30% of the women reported having entered a sexual relationship out of economic necessity and had increased odds of HIV infection, Odds Ratio (OR) 6.3 (P < 0.001). We postulate that due to recent economic hardship in rural Haiti, women are entering into sexual relationships out of economic necessity and that this trend is contributing to the growing HIV epidemic. We recommend STD prevention and development programs that target young people and economically disadvantaged women.


Assessing Fluid Resuscitation in Adults with Sepsis Who Are Not Mechanically Ventilated: a Systematic Review of Diagnostic Test Accuracy Studies.
Fluid resuscitation is a widely used intervention that is mandated in the management of sepsis. While its use can be life-saving, its overuse is associated with harm. Despite this, the best means of assessing a need for fluid resuscitation in an acute medical setting is unclear. To assess studies of diagnostic tests that identify the need for fluid resuscitation in adults with sepsis, as defined by the presence of fluid responsiveness. Protocol registration was performed in advance (PROSPERO:CRD42017048651). Research database searches were performed alongside additional searches to identify grey literature. Diagnostic test accuracy studies that assessed any fluid assessment tool were identified independently by two authors, before data extraction and quality assessments were performed. Adults with sepsis, without intensive care organ support, who would be appropriate for admission to an acute medical unit. Of the 26,841 articles that were screened, 14 studies were identified for inclusion, involving a combined total of 594 patients. Five categories of index test were identified: inferior vena cava collapsibility index (IVCCI), haemodynamic change with passive leg raise, haemodynamic change with respiration, haemodynamic change with intravenous fluid administration, and static assessment tools. Due to the high level of clinical heterogeneity affecting all aspects of study design, quantitative analysis was not feasible. There was a lack of consensus on reference tests to determine fluid responsiveness. While fluid resuscitation is considered a key part of the management of sepsis, evidence to support fluid assessment in awake adults is lacking. This review has highlighted a number of research recommendations that should be addressed as a matter of urgency if patient harm is to be avoided.


Perceptions of surgeons on surgical antibiotic prophylaxis use at an urban tertiary hospital in Tanzania.
Surgical Site Infections are a major cause of morbidity and mortality among operated patients. In spite of the accessibility of universal and national guidelines for surgical prophylaxis, recent studies surveying the present routine of prophylaxis have demonstrated overutilization of a wide range antibacterial medication for a single patient. Few studies have shown qualitatively factors influencing this and perceptions of surgeons on surgical antibiotic prophylaxis use. Unfortunately, none of these studies have been done in Tanzania. To describe the perceptions of surgeons on surgical antibiotic prophylaxis use at an urban tertiary hospital. A qualitative study involving in-depth interviews with surgeons was conducted in English by the primary investigator. The interviews were audio-recorded and transcribed verbatim. Systematic text condensation by Malterud was used for data analysis. Fourteen surgeons and obstetrics and gynaecologists participated. Their perceptions were summarized into three main categories: Inadequate data to support practice; one who sees the patient decides the antibiotic prophylaxis; prolonged antibiotic use for fear of unknown. The participants perceived that choice of antibiotic should be based on local hospital data for bacterial resistance pattern, however the hospital guidelines and data for surgical site infection rates are unknown. Fear of getting infection and anticipating complications led to prolonged antibiotics use. The study provides an understanding of surgical antibiotic prophylaxis use and its implementation challenges. This was partly expressed by unavailability of local data and guidelines to enhance practice. To improve this, there is a need of guidelines that incorporates local resistance surveillance data and enhanced antibiotic stewardship programmes. A strong consideration should be placed into ways to combat the fears of surgeons for complications, as these significantly affect the current practise with use of surgical antibiotic prophylaxis.


Development of a Physiologically Based Pharmacokinetic Model to Predict Disease-Mediated Therapeutic Protein-Drug Interactions: Modulation of Multiple Cytochrome P450 Enzymes by Interleukin-6.
Disease-mediated therapeutic protein-drug interactions have recently gained attention from regulatory agencies and pharmaceutical industries in the development of new biological products. In this study, we developed a physiologically based pharmacokinetic (PBPK) model using SimCYP to predict the impact of elevated interleukin-6 (IL-6) levels on cytochrome P450 (CYP) enzymes and the treatment effect of an anti-IL-6 monoclonal antibody, sirukumab, in patients with rheumatoid arthritis (RA). A virtual RA patient population was first constructed by incorporating the impact of systemic IL-6 level on hepatic and intestinal expression of multiple CYP enzymes with information from in vitro studies. Then, a PBPK model for CYP enzyme substrates was developed for healthy adult subjects. After incorporating the virtual RA patient population, the PBPK model was applied to quantitatively predict pharmacokinetics of multiple CYP substrates in RA patients before and after sirukumab treatment from a clinical cocktail drug interaction study. The results suggested that, compared with observed clinical data, changes in systemic exposure to multiple CYP substrates by anti-IL-6 treatment in virtual RA patients have been reasonably captured by the PBPK model, as manifested by modulations in area under plasma concentration versus time curves for midazolam, omeprazole, S-warfarin, and caffeine. This PBPK model reasonably captured the modulation effect of IL-6 and sirukumab on activity of CYP3A, CYP2C9, CYP2C19, and CYP1A2 and holds the potential to be utilized to assess the modulation effect of sirukumab on the metabolism and pharmacokinetics of concomitant small-molecule drugs in RA patients.


Traumatic brain injury among female veterans: a review of sex differences in military neurosurgery.
The impact of traumatic brain injury (TBI) has been demonstrated in various studies with respect to prevalence, morbidity, and mortality data. Many of the patients burdened with long-term sequelae of TBI are veterans. Although fewer in number, female veterans with TBI have been suggested to suffer from unique physical, mental, and social challenges. However, there remains a significant knowledge gap in the sex differences in TBI. Increased female representation in the military heralds an increased risk of TBI for female soldiers, and medical professionals must be prepared to address the unique health challenges in the face of changing demographics among the veteran TBI population. In this review, the authors aimed to present the current understanding of sex differences in TBI in the veteran population and suggest directions for future investigations.


Transcatheter aortic valve replacement in patients with severe aortic stenosis who are at high risk for surgical complications: summary assessment of the California Technology Assessment Forum.
The California Technology Assessment Forum is dedicated to assessment and public reporting of syntheses of available data on medical technologies. In this assessment, transcatheter aortic valve replacement (TAVR) was evaluated for patients with severe aortic stenosis (AS) who are at high risk for complications. In this assessment, 5 criteria were used: Regulatory approval, sufficient scientific evidence to allow conclusions on effectiveness, evidence that the technology improves net health outcomes, evidence that the technology is as beneficial as established methods, and availability of the technology outside investigational settings. In this assessment, all 5 criteria were judged to have been met. The primary benefit of TAVR is the ability to treat AS in patients who would otherwise be ineligible for surgical aortic valve replacement. It may also be useful for patients at high surgical risk by potentially reducing periprocedural complications and avoiding the morbidity and recovery from undergoing heart surgery. Potential harms include the need for conversion to an open procedure, perioperative death, myocardial infarction, stroke, bleeding, valve embolization, aortic regurgitation, heart block that requires a permanent pacemaker, renal failure, pulmonary failure, and major vascular complications such as cardiac perforation or arterial dissection. Potential long-term harms include death, stroke, valve failure or clotting, and endocarditis. As highlighted at the February 2012 California Technology Assessment Forum meeting, the dispersion of this technology to new centers across the United States must proceed with careful thought given to training and proctoring multidisciplinary teams to become new centers of excellence. TAVR is a potentially lifesaving procedure that may improve quality of life for patients at high risk for surgical AVR. However, attention needs to be paid to appropriate patient selection, their preoperative evaluation, surgical techniques, and postoperative care to preserve and improve on the results attained in the Placement of Aortic Transcatheter Valve trial. Specialty societies are collaborating to ensure that this happens in a rational and comprehensive manner.


Targeting Microbiota: What Do We Know about It at Present?
The human microbiota is a variety of different microorganisms. The composition of microbiota varies from host to host, and it changes during the lifetime. It is known that microbiome may be changed because of a diet, bacteriophages and different processes for example, such as inflammation. Like all other areas of medicine, there is a continuous growth in the area of microbiology. Different microbes can reside in all sites of a human body, even in locations that were previously considered as sterile; for example, liver, pancreas, brain and adipose tissue. Presently one of the etiological factors for liver disease is considered to be pro-inflammatory changes in a host's organism. There are lot of supporting data about intestinal dysbiosis and increased intestinal permeability and its effect on development of liver disease pointing to the gut-liver axis. The gut-liver axis affects pathogenesis of many liver diseases, such as chronic hepatitis B, chronic hepatitis C, alcoholic liver disease, non-alcoholic liver disease, non-alcoholic steatohepatitis, liver cirrhosis and hepatocellular carcinoma. Gut microbiota has been implicated in the regulation of brain health, emphasizing the gut-brain axis. Also, experiments with mice showed that microorganisms have significant effects on the blood-brain barrier integrity. Microbiota can modulate a variety of mechanisms through the gut-liver axis and gut-brain axis. Normal intestinal flora impacts the health of a host in many positive ways, but there is now significant evidence that intestinal microbiota, especially altered, have the ability to impact the pathologies of many diseases through different inflammatory mechanisms. At this point, many of the pathophysiological reactions in case of microbial disbyosis are still unclear.


A prospective study of cutaneous abnormalities in patients with chronic kidney disease.
There are diverse ways in which the skin is affected by chronic kidney disease (CKD). Various specific and nonspecific skin abnormalities are observed in patients with CKD. The aim of the study was to document the prevalence of skin diseases that commonly occur in patients with CKD on medical treatment and dialysis. A total of 99 patients with CKD were examined for evidence of skin diseases. Ninety-six had at least one cutaneous abnormality attributable to CKD. The most prevalent finding was xerosis (66.7%), followed by pallor (45.45%), pruritus (43.4%), and cutaneous pigmentation (32.3%). Other cutaneous manifestations included dermatitis (27.27%); Kyrle's disease (17.17%); fungal (8.08%), bacterial (11.1%), and viral (5.05%) infections; purpura (10.1%); gynecomastia (4.04%); and yellow skin (5.05%). The common nail changes were half and half nails (36.36%) and onycholysis (13.13%). CKD is associated with various cutaneous abnormalities caused either by the disease or by treatment, the most common being xerosis and pruritus. The dermatologic complications can significantly impair the quality of life in certain individuals; therefore, earlier diagnosis and treatment is important to improve their quality of life.


Mental health needs and service utilization by Hispanic immigrants residing in mid-southern United States.
This study assessed mental health needs and service utilization patterns in a convenience sample of Hispanic immigrants. A total of 84 adult Hispanic participants completed a structured diagnostic interview and a semistructured service utilization interview with trained bilingual research assistants. In the sample, 36% met diagnostic criteria for at least one mental disorder. Although 42% of the sample saw a physician in the prior year, mental health services were being rendered primarily by religious leaders. The most common barriers to service utilization were cost (59%), lack of health insurance (35%), and language (31%). Although more women than men met criteria for a disorder, service utilization rates were comparable. Participants with a mental disorder were significantly more likely to have sought medical, but not psychiatric, services in the prior year and faced significantly more cost barriers than participants without a mental disorder. Findings suggest that Hispanic immigrants, particularly those with a mental illness, need to access services but face numerous systemic barriers. The authors recommend specific ways to make services more affordable and linguistically accessible.


The Association Between Toddlers' Temperament and Well-Being in Norwegian Early Childhood Education and Care, and the Moderating Effect of Center-Based Daycare Process Quality.
Children who experience well-being are engaging more confidently and positively with their caregiver(s) and peers, which helps them to profit more from available learning opportunities and support current and later life outcomes. The goodness-of-fit theory suggests that children's well-being might be a result of the interplay between their temperament and the environment. However, there is a lack of studies that examined the association between children's temperament and well-being in early childhood education and care (ECEC), and whether this association is affected by ECEC process quality. Using a multilevel random coefficient approach, this study examines the association between toddlers' (N = 1,561) temperament (shyness, emotionality, sociability, and activity) and well-being in Norwegian ECEC and investigates whether process quality moderates this association. Results reveal an association between temperament and well-being. Staff-child conflict moderates the association between shyness and well-being, and between activity and well-being. Moreover, high emotional behavioral support moderates the association between activity and well-being. Extra attention should be paid by the staff to these children's needs.


Paroxysmal non-kinesigenic dyskinesia: pathophysiological investigations.
The paroxysmal dyskinesias constitute a heterogeneous group of disorders whose pathophysiologies and nosologies are still generally unknown. In a boy with severe, paroxysmal non-kinesigenic dyskinesia (PNKD), an invasive video-electrographic study demonstrated that his PNKD did not originate from the cortex, while a discharge was registered from the caudate nuclei. An 18FDG PET scan failed to show metabolic anomalies. A 18FDOPA and a 11C raclopride PET scans revealed a marked reduction in the density of presynaptic dopa decarboxylase activity in the striatum, together with an increased density of postsynaptic dopamine D2 receptors. These findings may suggest a chronic upregulation of postsynaptic dopa receptors, either because of an increase in their numbers or changes in their affinity. It remains unknown if these are secondary to reduced dopamine synthesis or altered release, or if the changes in striatal dopamine receptor binding are a primary or a compensatory mechanism.


Tasks Driven by Perceptual Information Do Not Recruit Sustained BOLD Activity in Cingulo-Opercular Regions.
Sustained blood oxygen level dependent (BOLD) signal in the dorsal anterior cingulate cortex/medial superior frontal cortex (dACC/msFC) and bilateral anterior insula/frontal operculum (aI/fO) is found in a broad majority of tasks examined and is believed to function as a putative task set maintenance signal. For example, a meta-analysis investigating task-control signals identified the dorsal anterior cingulate cortex and anterior insula as exhibiting sustained activity across a variety of task types. Re-analysis of tasks included in that meta-analysis showed exceptions, suggesting that tasks where the information necessary to determine a response was present in the stimulus (i.e., perceptually driven) does not show strong sustained cingulo-opercular activity. In a new experiment, we tested the generality of this observation while addressing alternative explanations about sustained cingulo-opercular activity (including task difficulty and verbal vs. non-verbal task demands). A new, difficult, perceptually driven task was compared with 2 new tasks that depended on information beyond that provided by the stimulus. The perceptually driven task showed a lack of cingulo-opercular activity in contrast to the 2 newly constructed tasks. This finding supports the idea that sustained cingulo-opercular activity contributes to maintenance of task set in only a subset of tasks.


A hypothesis: Potential contributions of metals to the pathogenesis of pulmonary artery hypertension.
Pulmonary artery hypertension (PAH) is characterized by vasoconstriction and vascular remodeling resulting in both increased pulmonary vascular resistance (PVR) and pulmonary artery pressure (PAP). The chronic and high-pressure stress experienced by endothelial cells can give rise to inflammation, oxidative stress, and infiltration by immune cells. However, there is no clearly defined mechanism for PAH and available treatment options only provide limited symptomatic relief. Due to the far-reaching effects of metal exposures, the interaction between metals and the pulmonary vasculature is of particular interest. This review will briefly introduce the pathophysiology of PAH and then focus on the potential roles of metals, including essential and non-essential metals in the pathogenic process in the pulmonary arteries and right heart, which may be linked to PAH. Based on available data from human studies of occupational or environmental metal exposure, including lead, antimony, iron, and copper, the hypothesis of metals contributing to the pathogenesis of PAH is proposed as potential risk factors and underlying mechanisms for PAH. We propose that metals may initiate or exacerbate the pathogenesis of PAH, by providing potential mechanism by which metals interact with hypoxia-inducible factor and tumor suppressor p53 to modulate their downstream cellular proliferation pathways. These need further investigation. Additionally, we present future research directions on roles of metals in PAH.


Orthogonal view analysis for evaluating the femoral component position of total hip implants in dogs using postoperative radiographs.
To show the ability to obtain repeatable, accurate, quantitative data to assess the position of the femoral component of canine total hip implants using postoperative radiographs. Cadaveric study and clinical trial. Five cadaveric canine femurs and 4 patients. Femoral implants were placed into 5 cadaveric canine femurs in predetermined locations. Orthogonal radiographs were taken of each femur at 10 degrees intervals as they were moved through a natural range of hip motion. An assessment of implant position was made with each set of radiographs and analyzed to observe significant inconsistencies in the values obtained when the femur was repositioned. Three investigators positioned and radiographed the hips of 4 client-owned dogs that had total hip arthroplasty (THA). Each investigator obtained a set of orthogonal radiographic views from which implant position was measured. The measurements obtained from each investigator's set of radiographs were statistically analyzed to evaluate for interobserver differences. The methods described indicate that consistent data regarding femoral implant placement can be obtained from postoperative radiographs of THA patients. Under most circumstances, change in position of the femur did not significantly affect femoral implant measurements. There were no significant differences noted among values obtained by different investigators. Measurements based on the femoral and implant axes obtained from orthogonal radiographic views of the femur provide a means for obtaining accurate and consistent quantitative data regarding femoral implant position using postoperative radiographs of canine THA patients. Acquisition of quantitative information about femoral implant position using postoperative radiographs will facilitate development of a readily available data source. This information, attainable in a clinical setting, may help identify elements of implant position that are important in determining the clinical outcome of THA in dogs.


Genetic fusion of an alpha-subunit gene to the follicle-stimulating hormone and chorionic gonadotropin-beta subunit genes: production of a bifunctional protein.
The human glycoprotein hormones, hCG, TSH, LH, and FSH, are composed of a common alpha-subunit assembled to a hormone-specific beta-subunit. The subunits combine noncovalently early in the secretory pathway and exist as heterodimers but not as multimers. LH/FSH are synthesized in the pituitary gonadotrophs, and several of the alpha-subunit sequences required for association with either the LHbeta or FSHbeta subunits are different. Thus, it is intriguing that no ternary complexes are observed for LH and FSH in vivo (e.g. two different beta-assembled to a single alpha-subunit). To examine whether the alpha-subunit can interact with more than one beta-subunit, and to study the conformational relationships between the ligand and the receptor, we constructed a vector encoding two tandemly arranged beta-subunits fused to a single alpha-subunit gene (FSHbeta-CGbeta-alpha). This approach permitted structure-function analyses of alpha/beta domain complexes without the possibility of subunit dissociation. We reported previously that the CGbeta or FSHbeta subunit gene can be genetically fused to the alpha-gene and the resulting single chains (CGbetaalpha and FSHbetaalpha, respectively) were biologically active. Here we demonstrate that a triple-domain single chain bearing the configuration FSHbeta-CGbeta-alpha is efficiently secreted from transfected Chinese hamster ovary (CHO) cells and exhibits high-affinity receptor binding to both FSH and LH/hCG receptors, comparable to the native heterodimers. These results indicate that the alpha-subunit can interact with each beta-subunit in the same complex and that an alpha-domain fused to a beta-domain can still interact with an additional beta-subunit. The data also demonstrate the remarkable flexibility of the receptor to accommodate the increased bulkiness of the triple-domain ligand. In addition, the formation of intrachain FSH- and CG-like complexes observed in a triple-domain single chain suggests that the alpha-subunit can resonate, i.e. shuttle between alpha-beta heterodimeric intermediates during the early stages of synthesis and accumulation in the endoplasmic reticulum. Such model compounds could be useful as substrates to generate a new class of analogs in which the ratio of the LH/FSH activity is varied. This could aid in the design of analogs that could be used to mimic the in vivo hormonal profiles.


The Implementation of Mindfulness-Based, Trauma-Informed Parent Education in an Underserved Latino Community: The Emergence of a Community Workforce.
In this practice-oriented program review, a mindfulness-based, trauma-informed parent intervention, called Safe, Secure and Loved™ (SSL), designed to strengthen nurturing parenting and children's resilience, was implemented in an underserved Latino community. Across 5 years, a volunteer community workforce of promotoras transformed an academic-community research partnership into a community-led program partnership and established sustainable agency parent education programming. To better understand this transformation, we used a modified implementation science (IS) framework to structure interviews from members of the academic-community research partnership. Findings suggest that the commitment and cultural expertise of the volunteer community workforce acted as the major leadership drivers to create the community-led program partnership. Employing mindfulness-based, trauma-informed parent education designed to promote nurturing parenting and children's resilience may be an effective training model to engage and mobilize a volunteer community workforce from an underserved community.


Development of Limited-Angle Iterative Reconstruction Algorithms with Context Encoder-Based Sinogram Completion for Micro-CT Applications.
Limited-angle iterative reconstruction (LAIR) reduces the radiation dose required for computed tomography (CT) imaging by decreasing the range of the projection angle. We developed an image-quality-based stopping-criteria method with a flexible and innovative instrument design that, when combined with LAIR, provides the image quality of a conventional CT system. This study describes the construction of different scan acquisition protocols for micro-CT system applications. Fully-sampled Feldkamp (FDK)-reconstructed images were used as references for comparison to assess the image quality produced by these tested protocols. The insufficient portions of a sinogram were inpainted by applying a context encoder (CE), a type of generative adversarial network, to the LAIR process. The context image was passed through an encoder to identify features that were connected to the decoder using a channel-wise fully-connected layer. Our results evidence the excellent performance of this novel approach. Even when we reduce the radiation dose by 1/4, the iterative-based LAIR improved the full-width half-maximum, contrast-to-noise and signal-to-noise ratios by 20% to 40% compared to a fully-sampled FDK-based reconstruction. Our data support that this CE-based sinogram completion method enhances the efficacy and efficiency of LAIR and that would allow feasibility of limited angle reconstruction.


Effect of parental donor sex on rejection in pediatric renal transplantation: a report of the North American Pediatric Renal Transplant Cooperative Study.
Using the North American Pediatric Renal Transplant Cooperative Study (NAPRTCS) database, we performed a retrospective cohort study of 1,552 pediatric renal transplant patients who had received a graft from a biological parent to determine if parental donor sex influences the development of rejection. There were 102/675 (15.1%) graft failures in paternal grafts compared to 144/877 (16.4%) graft failures in maternal grafts. Overall graft survival (p=0.48) and time to first rejection (p>0.9) were not different in patients receiving paternal versus maternal grafts. The overall frequency of graft loss to rejection was also not different. However, maternal donation was associated with a significantly longer time to first rejection in patients less than one year of age at the time of transplantation (p=0.01). Time to first rejection was not different between maternal and paternal grafts in older recipients. In summary, the present study did not demonstrate a difference in graft survival between maternal and paternal donations, but the youngest patients may experience a longer time to first rejection with maternal donation. The number of young patients is small, however, and further data are necessary to confirm this observation.