Evaluation of a hepatitis C clinical care coordination programme's effect on treatment initiation and cure: A surveillance-based propensity score matching approach.

Hepatitis C (HCV) is a viral infection that if left untreated can severely damage the liver. Project INSPIRE was a 3 year HCV care coordination programme in New York City (NYC) that aimed to address barriers to treatment initiation and cure by providing patients with supportive services and health promotion. We examined whether enrolment in Project INSPIRE was associated with differences in HCV treatment and cure compared with a demographically similar group not enrolled in the programme. INSPIRE participants in 2015 were matched with a cohort of HCV-infected persons identified in the NYC surveillance registry, using full optimal matching on propensity scores and stratified by INSPIRE enrolment status. Conditional logistic regression was used to assess group differences in the two treatment outcomes. Two follow-up sensitivity analyses using individual pair-matched sets and the full unadjusted cohort were also conducted. Treatment was initiated by 72% (790/1130) of INSPIRE participants and 36% (11 960/32 819) of study-eligible controls. Among initiators, 65% (514/790) of INSPIRE participants compared with 47% (5641/11 960) of controls achieved cure. In the matched analysis, enrolment in INSPIRE increased the odds of treatment initiation (OR: 5.25, 95% CI: 4.47-6.17) and cure (OR: 2.52, 95% CI: 2.00-3.16). Results from the sensitivity analyses showed agreement with the results from the full optimal match. Participation in the HCV care coordination programme significantly increased the probability of treatment initiation and cure, demonstrating that care coordination for HCV-infected individuals improves treatment outcomes.

Direct Screening of Blood by PCR and Pyrosequencing for a 16S rRNA Gene Target from Emergency Department and Intensive Care Unit Patients Being Evaluated for Bloodstream Infection.

Here we compared the results of PCR/pyrosequencing to those of culture for detecting bacteria directly from blood. DNA was extracted from 1,130 blood samples from 913 patients suspected of bacteremia (enrollment criteria were physician-ordered blood culture and complete blood count [CBC]), and 102 controls (healthy blood donors). Real-time PCR assays for beta-globin and Universal 16S rRNA gene targets were performed on all 1,232 extracts. Specimens identified by Universal 16S rRNA gene PCR/pyrosequencing as containing staphylococci, streptococci, or enteric Gram-negative rods had target-specific PCR/pyrosequencing performed. Amplifiable beta-globin (melting temperature [Tm], 87.2°C ± 0.2°C) occurred in 99.1% (1,120/1,130) of patient extracts and 100% (102/102) of controls. Concordance between PCR/pyrosequencing and culture was 96.9% (1,085/1,120) for Universal 16S rRNA gene targets, with positivity rates of 9.4% (105/1,120) and 11.3% (126/1,120), respectively. Bacteria cultured included staphylococci (59/126, 46.8%), Gram-negative rods (34/126, 27%), streptococci (32/126, 25.4%), and a Gram-positive rod (1/126, 0.8%). All controls screened negative by PCR/pyrosequencing. Clinical performance characteristics (95% confidence interval [CI]) for Universal 16S rRNA gene PCR/pyrosequencing included sensitivity of 77.8% (69.5 to 84.7), specificity of 99.3% (98.6 to 99.7), positive predictive value (PPV) of 93.3% (86.8 to 97.3), and negative predictive value (NPV) of 97.2% (96.0 to 98.2). Bacteria were accurately identified in 77.8% (98/126) of culture-confirmed sepsis samples with Universal 16S PCR/pyrosequencing and in 76.4% (96/126) with follow-up target-specific PCR/pyrosequencing. The initial PCR/pyrosequencing took ∼5.5 h to complete or ∼7.5 h when including target-specific PCR/pyrosequencing compared to 27.9 ± 13.6 h for Gram stain or 81.6 ± 24.0 h for phenotypic identification. In summary, this molecular approach detected the causative bacteria in over three-quarters of all culture-confirmed cases of bacteremia directly from blood in significantly less time than standard culture but cannot be used to rule out infection.

Perceived energy compensation following various sports: an age and sex comparison. Preliminary observations.

Following periods of physical activity, it is not uncommon for exercisers to increase their energy intake as a reward deemed 'earned'. Consumers' awareness of the energy within food and expended from exercise has previously been found to be limited. Therefore, the aim was to investigate whether habitual exercisers (50 adults and 49 children from 5 sports clubs) were able to conceptualise the energy expenditure (EE), following 1 h of their regular sports training, into a quantifiable amount of perceived energy compensation (PEC) in the form of food (chocolate) or drink (sports drink). Mean percentage accuracy for the PEC against EE matched <30% (± 29%), a significant underestimation irrespective of sex or sport. Percentage accuracy failed to significantly correlate to age. These findings indicate a necessity to improve nutrition education surrounding the energy costs of exercise relative to the energy contained within foods/drinks for both adults and children.

The soy isoflavone genistein inhibits the reduction in Achilles tendon collagen content induced by ovariectomy in rats.

The objective of this study was to evaluate the effects of genistein and moderate intensity exercise on Achilles tendon collagen and cross-linking in intact and ovariectomized (OVX) female Sprague-Dawley rats. Rats were separated into eight groups (n = 9/group): intact or OVX, treadmill exercised or sedentary, genistein-treated (300 mg/kg/day) or vehicle. After 6 weeks, tendons were assayed for the collagen-specific amino acid hydroxyproline and hydroxylyslpyridinoline (HP). Collagen content was not influenced by exercise (P = 0.40) but was lower (P < 0.001) in OVX-vehicle rats compared with intact vehicle rats (OVX: 894 ± 35 µg collagen/mg dry weight; intact: 1185 ± 72 µg collagen/mg dry weight). In contrast, collagen content in OVX rats treated with genistein was greater (P = 0.010, 1198 ± 121 µg collagen/mg dry weight) when compared with untreated rats and was not different from intact rats (P = 0.89). HP content was lower in OVX genistein-treated rats when compared with intact genistein-treated rats, but only within the sedentary animals (P = 0.05, intact-treated: 232 ± 39 mmol/mol collagen; OVX-treated: 144 ± 21 mmol/mol collagen). Our findings suggest that ovariectomy leads to a reduction in tendon collagen, which is prevented by genistein. HP content, however, may not have increased in proportion to the addition of collagen. Genistein may be useful for improving tendon collagen content in conditions of estrogen deficiency.

Improving memory span in children with Down syndrome.

BACKGROUND: Down syndrome (DS) is characterized by impaired memory span, particularly auditory verbal memory span. Memory span is linked developmentally to several language capabilities, and may be a basic capacity that enables language learning. If children with DS had better memory span, they might benefit more from language intervention. The present study evaluates a home-based parent-implemented intervention designed to improve auditory memory span in children with DS. METHOD: Sixteen children with DS, age 6-14, completed one or two 3-month periods of memory training using overt cumulative rehearsal and auditory-only procedures. Children were divided into two groups. Group 1 completed 3 months of memory training followed by 3 months of visual activities, followed by three more months of memory training. Group 2 had the opposite schedule. Before and after each 3-month period, children came into the laboratory for memory assessment. RESULTS: Children improved in training sessions and a small amount on digit span, the primary proximal outcome assessment measure. Digit span improvement was linked to the memory training, as indicated by control comparisons and correlations. Improvement was correlated with language comprehension and verbal working memory, but not with non-verbal ability, age or home/behavioural variables. No improvement was evident on distal outcome measures (sentence memory and verbal working memory); however, a phonological similarity effect emerged coincidence with memory training, suggesting increased use of phonological codes in memory. CONCLUSIONS: Results suggest that some children with DS can improve their auditory verbal memory span with home-based rehearsal training, at least in limited ways. Children with good language and verbal working memory skills may be the best candidates for this type of training, even though they may show only small improvements. Still, small improvements in a severely impaired function are noteworthy, and justify further investigation.

Significance of non-calcified pulmonary nodules in patients with extrapulmonary cancers.

BACKGROUND: This study sought to determine the rate and patterns of malignancy in patients with extrapulmonary cancers and non-calcified pulmonary nodules, and to develop a statistical model to guide clinicians regarding choice of patients for diagnostic biopsy. METHOD: The medical records of 151 patients evaluated at the Memorial Sloan-Kettering Cancer Center between January 1999 and December 2001 for non-calcified pulmonary nodules were reviewed. Nodules were considered malignant based on the results of a diagnostic biopsy, and were considered benign if their appearance remained stable 2 years after the initial study, if they resolved, or if a biopsy showed a non-malignant condition. RESULTS: Sixty four of 151 patients (42%) were diagnosed with malignant nodules; 32 had newly diagnosed lung cancers, 28 had metastatic spread of their primary cancers, and four had lesions that were either new cancers or of undetermined aetiology. On univariate analysis the likelihood of malignancy increased with nodule size, tobacco exposure, and the finding of a solitary nodule. On multivariable analysis only nodule size and tobacco exposure were predictive of malignancy. The model had good predictive accuracy (area under the curve 0.751) but had insufficient discrimination for use as a clinical tool to determine which patients should undergo diagnostic biopsy. CONCLUSION: Nearly half the non-calcified pulmonary nodules identified in this series were malignant. Lung cancer was more common than metastatic disease. These findings support the need for close interval follow up and a low threshold for diagnostic biopsy in patients with extrapulmonary cancers and non-calcified pulmonary nodules. In smokers, such lesions should raise concern for lung cancer.

Leptin does not respond to 48 h fat deposition or mobilization in women.

OBJECTIVE: To test the hypothesis that acute responses of plasma leptin concentration to energy balance manipulation are mediated by fat flux. DESIGN: Ten healthy women aged 31-63 y, mass 48-113.5 kg, fat mass 8.5-62.5 kg, were studied for 3 days in a whole-body calorimeter on two occasions. After a control day (D1) during which energy balance was maintained, diet was manipulated to induce fat deposition (FD) or mobilization (FM) of 50 g/day for 2 days (D2 & D3). A difference totalling of 194+/-18.6 g fat was achieved between manipulations without significant effects on carbohydrate or protein balance. Fasting plasma leptin was measured on D2 and D4. RESULTS: After the control day plasma leptin concentration averaged 19.01+/-9.8 ng/ml, and was found to be linearly related to body fat mass. After 2 days manipulation of fat balance, leptin concentrations were 21.4+/-10.3 ng/ml (FD) and 21.2+/-11.3 ng/ml (FM). There was no significant difference between treatments in either control day or postmanipulation leptin concentrations, nor did the treatments induce any differences in glucose or insulin concentration responses. CONCLUSION: Although in states of energy balance leptin concentration is linearly related to fat mass, acute modulation of leptin concentration during energy imbalance is not mediated by fat flux.

Role of importin-beta in coupling Ran to downstream targets in microtubule assembly.

The guanosine triphosphatase Ran stimulates assembly of microtubule asters and spindles in mitotic Xenopus egg extracts. A carboxyl-terminal region of the nuclear-mitotic apparatus protein (NuMA), a nuclear protein required for organizing mitotic spindle poles, mimics Ran's ability to induce asters. This NuMA fragment also specifically interacted with the nuclear transport factor, importin-beta. We show that importin-beta is an inhibitor of microtubule aster assembly in Xenopus egg extracts and that Ran regulates the interaction between importin-beta and NuMA. Importin-beta therefore links NuMA to regulation by Ran. This suggests that similar mechanisms regulate nuclear import during interphase and spindle assembly during mitosis.

Nuclear import in digitonin-permeabilized cells.

Import of proteins into the nucleus of a cell is a complex process that can be reconstituted in vitro. Digitonin-permeabilized cells are washed free of cytosolic factors to provide competent nuclei. Cytosolic factors for import are provided by an extract of Xenopus ovarian cells. Fluorochrome-conjugated probes, cloned proteins fused to green fluorescent protein, or antibodies to the protein of interest are used to visualize nuclear import. The system can also be used to identify nuclear localization sequences (NLS) of imported proteins.

An efficient color representation for image retrieval.

A compact color descriptor and an efficient indexing method for this descriptor are presented. The target application is similarity retrieval in large image databases using color. Colors in a given region are clustered into a small number of representative colors. The feature descriptor consists of the representative colors and their percentages in the region. A similarity measure similar to the quadratic color histogram distance measure is defined for this descriptor. The representative colors can be indexed in the three-dimensional (3-D) color space thus avoiding the high-dimensional indexing problems associated with the traditional color histogram. For similarity retrieval, each representative color in the query image or region is used independently to find regions containing that color. The matches from all of the query colors are then combined to obtain the final retrievals. An efficient indexing scheme for fast retrieval is presented. Experimental results show that this compact descriptor is effective and compares favorably with the traditional color histogram in terms of overall computational complexity.

Selective disruption of nuclear import by a functional mutant nuclear transport carrier.

p10/NTF2 is a nuclear transport carrier that mediates the uptake of cytoplasmic RanGDP into the nucleus. We constructed a point mutant of p10, D23A, that exhibited unexpected behavior both in digitonin-permeabilized and microinjected mammalian cells. D23A p10 was markedly more efficient than wild-type (wt) p10 at supporting Ran import, but simultaneously acted as a dominant-negative inhibitor of classical nuclear localization sequence (cNLS)-mediated nuclear import supported by karyopherins (Kaps) alpha and beta1. Binding studies indicated that these two nuclear transport carriers of different classes, p10 and Kap-beta1, compete for identical and/or overlapping binding sites at the nuclear pore complex (NPC) and that D23A p10 has an increased affinity relative to wt p10 and Kap-beta1 for these shared binding sites. Because of this increased affinity, D23A p10 is able to import its own cargo (RanGDP) more efficiently than wt p10, but Kap-beta1 can no longer compete efficiently for shared NPC docking sites, thus the import of cNLS cargo is inhibited. The competition of different nuclear carriers for shared NPC docking sites observed here predicts a dynamic equilibrium between multiple nuclear transport pathways inside the cell that could be easily shifted by a transient modification of one of the carriers.

Interactions between physical activity and diet in the regulation of body weight.

The increasing worldwide prevalence of obesity suggests that there is a chronic positive energy balance. This situation implies poor coupling between energy intake and energy expenditure, but the contribution of each of these factors remains unclear. Epidemiological data suggests that physical activity has a role in the prevention of weight gain, although there is little evidence that it has a beneficial role in weight loss. High-fat diets have also been implicated, evidence from a variety of sources suggests that diets high in fat undermine appetite regulatory mechanisms. There has been much research to investigate the coupling between energy expenditure and energy intake in the short term in an attempt to elucidate some of the mechanisms involved. However, mechanisms regulating appetite are very complex, and it is currently unclear at which points physical activity and diet may have an influence.

The nuclear import of RCC1 requires a specific nuclear localization sequence receptor, karyopherin alpha3/Qip.

RCC1 is the only known guanine nucleotide exchange factor for the small GTPase Ran and is normally found inside the nucleus bound to chromatin. In order to analyze in more detail the nuclear import of RCC1, we created a fusion construct in which four IgG binding domains of protein A were fused to the amino terminus of human RCC1 (pA-RCC1). Surprisingly, we found that neither Xenopus ovarian cytosol nor a mixture of recombinant import factors (karyopherin alpha2, karyopherin beta1, Ran, and p10/NTF2) were able to support the import of pA-RCC1 into the nuclei of digitonin-permeabilized cells. Both, in contrast, were capable of supporting the import of a construct containing another classical nuclear localization sequence (NLS), glutathione S-transferase-green fluorescent protein-NLS. Subsequently, we found that only one of the NLS receptors, karyopherin alpha3 (Kapalpha3/Qip), would support significant nuclear import of pA-RCC1 in permeabilized cells, while members of the other two main classes, Kapalpha1 and Kapalpha2, would not. Accordingly, in vitro binding studies revealed that only Kapalpha3 showed significant binding to RCC1 (unlike Kapalpha1 and Kapalpha2) and that this binding was dependent on the basic amino acids present in the RCC1 NLS. In addition to Kapalpha3, we found that the nuclear import of pA-RCC1 also required both karyopherin beta1 and Ran.

Dopamine modulates acute responses to cocaine, nicotine and ethanol in Drosophila.

BACKGROUND: Drugs of abuse have a common property in mammals, which is their ability to facilitate the release of the neurotransmitter and neuromodulator dopamine in specific brain regions involved in reward and motivation. This increase in synaptic dopamine levels is believed to act as a positive reinforcer and to mediate some of the acute responses to drugs. The mechanisms by which dopamine regulates acute drug responses and addiction remain unknown. RESULTS: We present evidence that dopamine plays a role in the responses of Drosophila to cocaine, nicotine or ethanol. We used a startle-induced negative geotaxis assay and a locomotor tracking system to measure the effect of psychostimulants on fly behavior. Using these assays, we show that acute responses to cocaine and nicotine are blunted by pharmacologically induced reductions in dopamine levels. Cocaine and nicotine showed a high degree of synergy in their effects, which is consistent with an action through convergent pathways. In addition, we found that dopamine is involved in the acute locomotor-activating effect, but not the sedating effect, of ethanol. CONCLUSIONS: We show that in Drosophila, as in mammals, dopaminergic pathways play a role in modulating specific behavioral responses to cocaine, nicotine or ethanol. We therefore suggest that Drosophila can be used as a genetically tractable model system in which to study the mechanisms underlying behavioral responses to multiple drugs of abuse.

The control of gene expression by regulated nuclear transport.

Many proteins show distinct nuclear- and cytoplasmic-localization patterns. For proteins above the diffusion limit of the NPC, this localization is governed by the activity of NLSs and/or NESs contained in the protein. Structural modification of proteins can affect NLS and NES activities. Ligand binding, phosphorylation and proteolysis are each capable of modifying the nucleocytoplasmic distribution of proteins. In the case of transcription factors, control of these structural modifications affects access of the transcription factor to the chromatin. This management of cellular distribution, in turn, regulates gene expression.

Contribution of a sedentary lifestyle and inactivity to the etiology of overweight and obesity: current evidence and research issues.

PURPOSE: The etiology of overweight and obesity is clearly multifactorial, but ultimately it is determined by the long-term balance between energy intake and expenditure. This review will consider the effects on body weight and the risk of obesity of sedentary lifestyles, within the context of dietary habits. METHODS: The data from ecological, cross-sectional, and prospective studies that have assessed physical activity and dietary intake and their relationship to body weight were reviewed. RESULTS: Ecological analyses imply that the increase in the prevalence of obesity is more strongly related to lower levels of physical activity than higher energy intakes. However, there is a paucity of pertinent data from cross-sectional or prospective studies. There is some evidence that both a high proportion of dietary fat and low levels of physical activity may increase the likelihood of weight gain. However, even the most comprehensive studies are unable to account for more than a small proportion of the interindividual variance in weight gain, so it is difficult to usefully assess their relative importance. Furthermore, there are insufficient data that pertain to "sedentary lifestyles" to segregate any putative effect from a protective effect of exercise. All the data in this review is NHLBI Evidence category C. CONCLUSIONS: This review provides clear evidence that low levels of physical activity are associated with an increased risk of weight gain and obesity. On balance, the evidence is suggestive of a causal link, but the experimental designs are too weak is provide conclusive evidence. The potential effect of interactions between diet and activity have largely been ignored. To make progress in this area, a number of key issues need to be resolved with regard to the methodology, study design, and statistical analysis of prospective epidemiological studies. In the meantime, data need to be drawn from other sources, particularly those studies designed to elucidate the mechanism of action of diet and physical activity in the etiology of obesity, to establish rational interventions to guide public health policies.

Getting across the nuclear pore complex.

The nuclear pore complex (NPC) connects the cytoplasm and nucleus through the nuclear envelope and serves as the pipeline for moving material between the two compartments. Macromolecules that move through the NPC range in size from the very small (for example, ions and ATP) to the very large (for example, ribonucleoprotein particle complexes). Unlike translocation across other organelle membranes, proteins do not have to be unfolded to be transported through the NPC, and the NPC also routinely transports large, multicomponent substrates in both directions. This review focuses on current understanding of the different mechanisms by which macromolecules move across the NPC.

Engineered mutants in the switch II loop of Ran define the contribution made by key residues to the interaction with nuclear transport factor 2 (NTF2) and the role of this interaction in nuclear protein import.

Nuclear protein import requires a precisely choreographed series of interactions between nuclear pore components and soluble factors such as importin-beta, Ran, and nuclear transport factor 2 (NTF2). We used the crystal structure of the GDPRan-NTF2 complex to design mutants in the switch II loop of Ran to probe the contribution of Lys71, Phe72 and Arg76 to this interaction. X-ray crystallography showed that the F72Y, F72W and R76E mutations did not introduce major structural changes into the mutant Ran. The GDP-bound form of the switch II mutants showed no detectable binding to NTF2, providing direct evidence that salt bridges involving Lys71 and Arg76 and burying Phe72 are all crucial for the interaction between Ran and NTF2. Nuclear protein accumulation in digitonin-permeabilzed cells was impaired with Ran mutants deficient in NTF2 binding, confirming that the NTF2-Ran interaction is required for efficient transport. We used mutants of the yeast Ran homologue Gsp1p to investigate the effect of the F72Y and R76E mutations in vivo. Although neither mutant was viable when integrated into the genome as a single copy, yeast mildly overexpressing the Gsp1p mutant corresponding Ran F72Y on a centromeric plasmid were viable, confirming that this mutant retained the essential properties of wild-type Ran. However, yeast expressing the Gsp1p mutant corresponding to R76E to comparable levels were not viable, although strains overexpressing the mutant to higher levels using an episomal 2micrometers plasmid were viable, indicating that the R76E mutation may also have interfered with other interactions made by Gsp1p.

The porous-coated anatomic total hip prosthesis, inserted without cement. A prospective study with a minimum of ten years of follow-up.

One hundred consecutive primary total hip arthroplasties performed with use of a porous-coated anatomic total hip prosthesis, fixed without cement, in ninety-one patients were followed prospectively for a minimum of ten years. At the time of the most recent follow-up, twenty patients (twenty-three hips) had died and seventy-one patients (seventy-seven hips) were living. The average age of the living patients was sixty-six years (range, thirty-two to ninety-two years), and their average Harris hip score was 84 points (range, 33 to 100 points). Twelve percent (nine) of the seventy-seven hips were found to be associated with pain in the thigh when the patients were specifically questioned by the examiner. Eleven hips were revised during the follow-up period. Only the acetabular component was revised in six hips, only the femoral component was revised in one hip, and both the femoral and the acetabular components were revised in four hips. Of the ten acetabular revisions, one was performed because of acute dissociation of the component and eight, because of a combination of polyethylene wear, osteolysis, and loosening; the tenth acetabular revision consisted of exchange of the liner and curettage and bone-grafting of the osteolytic area. Of the five femoral revisions, two were performed because of loosening and three, because of extensive osteolysis of the proximal aspect of the femur. Including the revised components, twelve acetabular components and five femoral components had radiographic evidence of aseptic loosening. Acetabular osteolysis occurred in seventeen hips. Femoral osteolysis occurred in thirty-nine hips: in the proximal aspect of thirty-one hips, in the distal aspect of four, and in both the proximal and the distal aspect of four. The durability of the femoral fixation documented in this study is especially encouraging in view of the fact that this was our initial experience with devices fixed without cement and that a so-called first-generation femoral component was used. However, the study also demonstrated that not all acetabular components fixed without cement function well over the long term and that specific design considerations (adequate initial fixation, congruency between the liner and the shell, an optimum shell-liner capturing mechanism, and a smaller femoral head) are warranted.