Near-infrared bone densitometry: A feasibility study on distal radius measurement.
Osteoporosis, defined as decreased bone mineral density (BMD), poses patients in dangers for fracture risk and has become a major public health problem worldwide because of is associated morbidity, mortality and costs. Without doubt, early detection and timely intervention are important to successfully manage osteoporosis and its associated complications. The dual-energy x-ray absorptiometry (DXA) is the most popular and standard method to measure BMD. However, limitations including radiation exposure and availability restrict its application for osteoporosis screening among general population. In this study, we developed a simple method to detect human distal radius bone density based on near infrared (NIR) image system. Among 10 volunteers (including 5 young and 5 elderly participants) receiving bone density measurement using our NIR image system at the ultradistal part of bilateral distal radius, we demonstrated a strong correlation between the optical attenuation and BMD measured with DXA, which may facilitate predicting bone density status. We hope our potential NIR image system may open a new avenue for development of osteoporosis screening facilities and help in prevention of osteoporosis related fracture and its associated complications in the near future. Pearson's correlations between BMD values from the DXA and light intensity of NIR system.


Alternative splicing in the pore-forming region of shaker potassium channels.
We have cloned cDNAs for the shaker potassium channel gene from the spiny lobster Panulirus interruptus. As previously found in Drosophila, there is alternative splicing at the 5' and 3' ends of the coding region. However, in Panulirus shaker, alternative splicing also occurs within the pore-forming region of the protein. Three different splice variants were found within the P region, two of which bestow unique electrophysiological characteristics to channel function. Pore I and pore II variants differ in voltage dependence for activation, kinetics of inactivation, current rectification, and drug resistance. The pore 0 variant lacks a P region exon and does not produce a functional channel. This is the first example of alternative splicing within the pore-forming region of a voltage-dependent ion channel. We used a recently identified potassium channel blocker, kappa-conotoxin PVIIA, to study the physiological role of the two pore forms. The toxin selectively blocked one pore form, whereas the other form, heteromers between the two pore forms, and Panulirus shal were not blocked. When it was tested in the Panulirus stomatogastric ganglion, the toxin produced no effects on transient K+ currents or synaptic transmission between neurons.


Cardiopulmonary responses to treadmill and cycle ergometry exercise in patients with peripheral vascular disease.
Peripheral arterial disease (PAD) presenting as intermittent claudication (IC) is routinely assessed as the distance or time walked to the onset of pain, which often occurs before significant cardiopulmonary stress and is subject to confounding factors such as increased body mass and altered gait. Thus, where exercise-induced cardiovascular stress is desirable, such as in cardiac stress testing or clinical trials, an alternative modality of exercise is required. Cycling will circumvent several of the associated problems of treadmill walking and may provide an alternative preferable method of exercise, although there is limited information on the physiologic response of patients with PAD to cycling. This study compared the peak cardiorespiratory responses and the repeatability of cycling and treadmill exercise in patients with PAD. Ten men (mean age, 54 +/- 10 years) with stable IC completed two incremental exercise tests to the limit of tolerance on a treadmill and a cycle ergometer after familiarization with the outcome measures of exercise duration, work performed, respiratory gas exchange variables using continuous breath-by-breath measurement, heart rate, and ratings of perceived pain. Both methods of exercise assessment revealed high reproducibility in terms of absolute claudication time (treadmill, r = 0.95; cycle, r = 0.91), time to volitional fatigue (treadmill, r = 0.96; cycle, r = 0.91), and cardiopulmonary exercise responses such as the lactate threshold (treadmill, r = 0.95; cycle, r = 0.94), peak heart rate (treadmill, r = 0.94; cycle, r = 0.96), and peak oxygen uptake (treadmill, r = 0.98; cycle, r = 0.87). Cycling induced significantly higher cardiopulmonary responses (peak heart rate, peak carbon dioxide output, peak minute ventilation, and respiratory exchange ratio) than treadmill exercise. There was no difference in time to volitional fatigue or in absolute claudication time between exercise modalities. These results demonstrate that exercise testing using cycling offers an alternative method of cardiopulmonary testing for patients with IC that is equally reliable and reproducible to treadmill walking. Cycling may be preferable to treadmill exercise because it induces greater cardiopulmonary and metabolic responses and is better tolerated by patients.


Locomotion Inhibition of Cimex lectularius L. Following Topical, Sublethal Dose Application of the Chitin Synthesis Inhibitor Lufenuron.
To date, few studies have evaluated chitin synthesis inhibitors against bed bugs, although they would provide an alternative mode of action to circumvent insecticide resistance. Acute and sublethal effects of lufenuron were evaluated against two strains of the common bed bug. Combined acute and sublethal effects were used to calculate effective doses. The dose that was effective against 50% of Harlan strain bed bugs was 0.0081% (w/v), and was much higher against Bradenton strain bed bugs (1.11% w/v). Sublethal doses were chosen to determine the effect that leg abnormalities had on pulling force. Both Harlan and Bradenton strain bed bugs had significantly lower locomotion ability (p < 0.0001) following topical application of lufenuron. The observed sublethal effects that limit locomotion could prevent bed bugs from moving within a domicile and taking a blood meal, subsequently reducing a bed bug population over time.


The role of intracavitary therapies in the treatment of malignant pleural mesothelioma.
Surgery is one of the steps of multimodality approach for the treatment of MPM. Due to anatomical features, microscopically radical (R0) resection is never possible and a Macroscopic Complete Resection (R1) is considered the target for mesothelioma surgeons. Recently, intracavitary therapies have been described with the aim of extending the loco-regional effect of surgery. Different agents might be administered intrapleurally: chemotherapy drugs are the most widely used, but also photodynamic therapy (PDT) showed to lead to satisfactory long-term outcomes; furthermore, immunotherapies and gene therapies have been also reported. Despite promising results, no high-quality evidences are currently available and controlled randomized trials are required to establish the exact role of intracavitary therapies and to standardize the technique.


Topical Formulation Containing Naringenin: Efficacy against Ultraviolet B Irradiation-Induced Skin Inflammation and Oxidative Stress in Mice.
Naringenin (NGN) exhibits anti-inflammatory and antioxidant activities, but it remains undetermined its topical actions against ultraviolet B (UVB)-induced inflammation and oxidative stress in vivo. The purpose of this study was to evaluate the physicochemical and functional antioxidant stability of NGN containing formulations, and the effects of selected NGN containing formulation on UVB irradiation-induced skin inflammation and oxidative damage in hairless mice. NGN presented ferric reducing power, ability to scavenge 2,2'-azinobis (3-ethylbenzothiazoline- 6-sulfonic acid) (ABTS) and hydroxyl radical, and inhibited iron-independent and dependent lipid peroxidation. Among the three formulations containing NGN, only the F3 kept its physicochemical and functional stability over 180 days. Topical application of F3 in mice protected from UVB-induced skin damage by inhibiting edema and cytokine production (TNF-α, IL-1β, IL-6, and IL-10). Furthermore, F3 inhibited superoxide anion and lipid hydroperoxides production and maintained ferric reducing and ABTS scavenging abilities, catalase activity, and reduced glutathione levels. In addition, F3 maintained mRNA expression of cellular antioxidants glutathione peroxidase 1, glutathione reductase and transcription factor Nrf2 (nuclear factor erythroid 2-related factor 2), and induced mRNA expression of heme oxygenase-1. In conclusion, a formulation containing NGN may be a promising approach to protecting the skin from the deleterious effects of UVB irradiation.


Identifying the preferred subset of enzymatic profiles in nonlinear kinetic metabolic models via multiobjective global optimization and Pareto filters.
Optimization models in metabolic engineering and systems biology focus typically on optimizing a unique criterion, usually the synthesis rate of a metabolite of interest or the rate of growth. Connectivity and non-linear regulatory effects, however, make it necessary to consider multiple objectives in order to identify useful strategies that balance out different metabolic issues. This is a fundamental aspect, as optimization of maximum yield in a given condition may involve unrealistic values in other key processes. Due to the difficulties associated with detailed non-linear models, analysis using stoichiometric descriptions and linear optimization methods have become rather popular in systems biology. However, despite being useful, these approaches fail in capturing the intrinsic nonlinear nature of the underlying metabolic systems and the regulatory signals involved. Targeting more complex biological systems requires the application of global optimization methods to non-linear representations. In this work we address the multi-objective global optimization of metabolic networks that are described by a special class of models based on the power-law formalism: the generalized mass action (GMA) representation. Our goal is to develop global optimization methods capable of efficiently dealing with several biological criteria simultaneously. In order to overcome the numerical difficulties of dealing with multiple criteria in the optimization, we propose a heuristic approach based on the epsilon constraint method that reduces the computational burden of generating a set of Pareto optimal alternatives, each achieving a unique combination of objectives values. To facilitate the post-optimal analysis of these solutions and narrow down their number prior to being tested in the laboratory, we explore the use of Pareto filters that identify the preferred subset of enzymatic profiles. We demonstrate the usefulness of our approach by means of a case study that optimizes the ethanol production in the fermentation of Saccharomyces cerevisiae.


Monoclonal anti-IgG autoantibodies derived from lipopolysaccharide-activated spleen cells of 129/Sv mice.
In some colonies, 129/Sv mice produce, upon aging, a rheumatoid factor (RF) that is specific for mouse IgG2a but fails to react with IgG2a of the b allotype. It is not known whether this narrow specificity is due to the absence of other RF specificities in the repertoire of these mice or to the selective activation of the production of anti-IgG2a autoantibodies by a specific stimulus. To analyze the RF repertoire of 129/Sv mice, we have derived hybridomas from their spleen cells 3 d after an intraperitoneal injection of lipopolysaccharide. We have obtained 68 hybridomas secreting a monoclonal IgM with RF activity. This represents approximately 3 percent of the total number of hybridomas generated in four hybridizations. In addition, one monoclonal IgA RF was derived from unstimulated 129/Sv spleen cells. The specificities of these monoclonal RF were examined by testing their ability to bind to a panel of homologous and heterologous IgG preparations. The majority of the IgM RF reacted exclusively with a single mouse IgG subclass: 58 with IgG1, and 1 with IgG2a. Eight bound preferentially to IgG1 but cross-reacted to some extent with IgG2a and one was specific for a determinant shared by IgG1, IgG2a, and IgG3. The IgA RF derived from unstimulated spleen cells was primarily directed against IgG2a but cross- reacted somewhat with IgG2b. Identical results were obtained with two different monoclonal IgG1 and IgG2a proteins of the a allotype. No allotypic specificity was found for the anti-IgG1 RF, which all reacted well with IgG1 of the b allotype. In contrast, the IgM anti-IgG2a antibody exhibited such allotypic specificity because it failed to react with IgG2a of the b allotype. When tested on heterologous IgG preparations, all anti-IgG1 RF reacted better with rat IgG1, rat IgG2c, bovine IgG2, goat IgG2, and rabbit IgG than with mouse IgG1, demonstrating a particular homology between these Ig. On the basis of additional cross-reactions with other IgG, including rat IgG2a, rat IgG2b, bovine IgG1, goat IgG1, human IgG, and chicken IgG, seven different anti-IgG1 clonotypes could be identified. However, despite their heterogeneity, nearly all antigenic determinants recognized by anti-IgG 1 RF appeared to be located in the hinge region of the molecule. Total lack of binding to IgG1 Fab fragments was indeed observed, and only one antibody reacted with IgG1 Fc fragments. Unlike the anti-IgG1 RF, the IgM and the IgA anti-IgG2a antibodies did not cross-react with any heterologous IgG of the same panel. Altogether, t 1 different RF clonotypes could be distinguished on the basis of their fine specificity. The anti-IgG2a specificity of the RF spontaneously produced by 129/ Sv mice is thus not due to the absence of other RF specificities in the repertoire of these mice.


Psychiatric interviewing techniques II. Naturalistic study: eliciting factual information.
A naturalistic study was undertaken of 36 video and audio taped interviews made by 7 different psychiatric trainees. The interviews studied were those conducted in the ordinary course of clinic work for diagnostic and therapeutic planning purposes by trainees when first seeing the parent or parents of a child newly referred to a psychiatric clinic. It was found that a directive style with specific probes and requests for detailed descriptions was associated with the obtaining of better-quality factual information than that associated with a more free-style approach. Interviewers who talked less and who made use of open questions and checks tended to have more talkative informants. Double questions were liable to result in ambiguous answers, but multiple-choice questions did not appear to cause distortion and in some circumstances might be helpful.


Fetal MRI-calculated total lung volumes in the prediction of short-term outcome in giant omphalocele: preliminary findings.
To determine the value of fetal MRI-calculated total lung volumes (TLV) in the prediction of short-term outcome in patients with giant omphalocele (GO). We reviewed all cases of GO undergoing fetal MRI after 21 weeks' gestation and receiving postnatal care at our institution between 2003 and 2010. Observed/expected (O/E) TLV was calculated using age-matched TLV normograms [Radiology 2001;219:236-241]. Postnatal outcomes were stratified based on O/E TLV above or below 50% of expected. Seventeen GO cases fulfilled the entry criteria. The mean age at fetal MRI evaluation was 25.8 ± 4.8 weeks' gestation. The mean GO TLV (21.0 ± 13.2) was lower than age-matched population norms (p < 0.001), resulting in a mean O/E TLV of 52.3 ± 16.8%. The mean gestational age at delivery was 36.8 ± 1.6 weeks. Overall survival was 94%. Fourteen (88%) infants underwent staged reduction, and 2 underwent silver sulfadiazine treatment and delayed repair. Infants with ≤50% of predicted O/E TLV (n = 11, 65%) had lower Apgar scores at birth (p = 0.03), prolonged ventilatory support (p = 0.004), delayed oral intake (p = 0.03), and longer hospitalization (p = 0.03) compared to patients with ≥50% of expected O/E TLV. Two infants (both O/E TLV <50%) required tracheostomy placement. In the assessment of GO fetuses, MRI-based O/E TLV of <50% was predictive of increased postnatal morbidity.


Vibration compensating beam scanning interferometer for surface measurement.
Light beam scanning using a dispersive element and wavelength tuning is coupled with fiber-optic interferometry to realize a new surface measurement instrument. The instrument is capable of measuring nanoscale surface structures and form deviations. It features active vibration compensation and a small optical probe size that may be placed remotely from the main apparatus. Active vibration compensation is provided by the multiplexing of two interferometers with near common paths. Closed loop control of a mirror mounted on a piezoelectric transducer is used to keep the path length stable. Experiments were carried out to deduce the effectiveness of the vibration compensation and the ability to carry out a real measurement in the face of large environmental disturbance.


Gene synteny in species of Plasmodium.
We have attempted to establish the degree of linkage conservation between different species of the malaria parasite Plasmodium. Initially, the chromosome locations of 42 homologous genes were established in parasites from a rodent malaria species and the human malaria parasite P. falciparum. Of these genes, 26 appeared to be conserved within ten synteny groups between the two genomes. Several synteny groups were analysed further by long-range restriction mapping of digested chromosomes. Finally, a fine restriction map of one of the linkage groups was made from the rodent malaria parasites P. berghei and from P. falciparum and from the simian malaria parasite P. knowlesi. The fine-scale organisation of this linkage group appears to have remained intact among the three species, despite the evolutionary distance between them. This provides the first example of linkage conservation between the rodent, simian and human malaria species, which represent three different branches of the inferred phylogenetic tree of the genus Plasmodium.


Glutathione supplementation attenuates lipopolysaccharide-induced mitochondrial dysfunction and apoptosis in a mouse model of acute lung injury.
Acute lung injury (ALI) is a life threatening condition associated with hypoxemia, diffuse alveolar damage, inflammation, and loss of lung function. Lipopolysaccharide (LPS; endotoxin) from the outer membrane of Gram-negative bacteria is a major virulence factor involved in the development of ALI. The depletion of glutathione (GSH), an essential intra- and extra-cellular protective antioxidant, by LPS is an important event that contributes to the elevation in reactive oxygen species. Whether restoring GSH homeostasis can effectively ameliorate mitochondrial dysfunction and cellular apoptosis in ALI is unknown and therefore, was the focus of this study. In peripheral lung tissue of LPS-treated mice, hydrogen peroxide and protein nitration levels were significantly increased. Pre-treatment with GSH-ethyl ester (GSH-EE) prevented this increase in oxidative stress. LPS also increased the lactate/pyruvate ratio, attenuated SOD2 protein levels, and decreased ATP levels in the mouse lung indicative of mitochondrial dysfunction. Again, GSH-EE treatment preserved the mitochondrial function. Finally, our studies showed that LPS induced an increase in the mitochondrial translocation of Bax, caspase 3 activation, and nuclear DNA fragmentation and these parameters were all prevented with GSH-EE. Thus, this study suggests that GSH-EE supplementation may reduce the mitochondrial dysfunction associated with ALI.


Characterization of the facilitative effects of perinatal choline supplementation on timing and temporal memory.
Perinatal choline supplementation can improve performance on a variety of spatial memory tasks in adulthood. In order to extend these studies, we have investigated the effects of perinatal choline supplementation on the performance of a peak-interval timing task in which a 20 s temporal criterion was trained for a visual signal in adult (3-6 months old) rats. Following 5 weeks of baseline training, rats received systemic injections of nicotine (0.1, 0.2, or 0.4 mg/kg, s.c.) or saline prior to testing on the peak-interval timing task. The results indicated that perinatal choline supplementation increased rats' sensitivity to the 20 s temporal criterion during baseline training and facilitated the clock speed enhancing effects of nicotine during drug testing. The present study extends the types of long-term cognitive enhancement produced by perinatal choline supplementation to include the temporal processing domain and relates these effects to modifications in cholinergic function.


A comparative study on the leishmanicidal activity of the L-amino acid oxidases BjussuLAAO-II and BmooLAAO-II isolated from Brazilian Bothrops snake venoms.
This study aims to examine whether two L-amino acid oxidases isolated from Bothrops snake venom (SV-LAAOs) were cytotoxic to Leishmania (Leishmania) amazonensis and Leishmania (Viannia) braziliensis, two causative agents of leishmaniasis, which is an endemic disease in tropical and subtropical countries. The SV-LAAOs BjussuLAAO-II and BmooLAAO-II were isolated from Bothrops jararacussu and Bothrops moojeni venom, respectively, through a three-step chromatography process that used molecular exclusion, hydrophobic interaction, and affinity columns. BmooLAAO-II is a new SV-LAAO isoform that we isolated in this study. The purified BjussuLAAO-II and BmooLAAO-II had high L-amino acid oxidase-specific activity: 3481.17 and 4924.77 U/mg/min, respectively. Both SV-LAAOs were strongly cytotoxic to the two Leishmania species, even at low concentrations. At the same concentration, BjussuLAAO-II and BmooLAAO-II exerted different cytotoxic effects on the parasites. We reported for the first time that the SV-LAAOs suppressed cell proliferation and altered the mitochondrial membrane potential of the two Leishmania species. Surprisingly, BjussuLAAO-II increased the intracellular reactive oxygen species production only in L. (L.) amazonensis, while BmooLAAO-II increased the intracellular reactive oxygen species production only in L. (V.) braziliensis, indicating that these SV-LAAOs had a certain specificity of action.


Adverse events risk associated with bevacizumab addition to breast cancer chemotherapy: a meta-analysis.
Bevacizumab is a monoclonal antibody against vascular endothelial growth factor with the ability to increase progression-free survival in metastatic breast cancer (MBC). A systematic review and meta-analysis was conducted to determine the risk of the most clinically relevant adverse outcomes associated with the use of bevacizumab in the treatment of breast cancer. We included phase III clinical trials that used bevacizumab alone or in combination with chemotherapy as for MBC or locally recurrent. Statistical analyses were conducted to calculate summary odds ratio (OR) of the eight most relevant adverse outcomes related with bevacizumab. Five clinical trials were included in the meta-analysis. Summary odds ratios obtained showed a statistically significant bevacizumab-associated increased risk in four of the adverse outcomes studied: proteinuria (OR = 27.68), hypertension (OR = 12.76), left ventricular dysfunction (LVD) (OR = 2.25), and hemorrhagic events (OR = 4.07). No statistically significant differences were found for gastrointestinal (GI) perforation, vascular events, fatal events, or febrile neutropenia. Bevacizumab did increase the risk of LVD and hemorrhagic events. The addition of bevacizumab to chemotherapy in patients with metastatic breast cancer was not associated with a significant increase in grade ≥ 3 arterial or venous thromboembolic events, GI perforation, or fatal events.


Discriminative ability of the generic and condition specific Oral Impact on Daily Performance (OIDP) among adolescents with and without hypodontia.
The aims of this study were to (1) investigate to what extent the generic and condition specific (CS) forms of the oral impact of daily performance (OIDP) inventory discriminate between a group of patients with hypodontia and a group of patients having malocclusion, (2) assess the association of the generic and CS OIDP with severity and localisation of hypodontia, whilst adjusting for patients' age and sex. A total of 163 patients aged 10-17 years were included in a cross-sectional study. Two groups were investigated: 62 patients with non-syndromic hypodontia and 101 non-hypodontia patients. Both groups had a malocclusion of similar treatment need. All patients underwent a clinical and radiographic examination and completed a Norwegian version of the generic and the CS OIDP inventory. CS scores were established for impacts attributed to hypodontia. The mean number of missing teeth in the hypodontia group was 6.2. The prevalence of generic and CS oral impacts in the hypodontia group were 64% and 30%, and the corresponding rates in the non-hypodontia group were 62% and 10%. The generic OIDP did not discriminate between the two groups with respect to overall scores. The CS OIDP discriminated strongly between patients with and without hypodontia regarding problems with emotional status, showing teeth, social contact, speaking and carrying out work. Compared to the non-hypodontia group, patients with hypodontia, with severe hypodontia (≥ 6 missing teeth) and upper anterior hypodontia were respectively 3.4, 2.5 and 7.0 times more likely to report any oral impact attributed to small teeth, gaps between teeth and missing teeth. Hypodontia and malocclusion patients report a considerable burden of oral impacts. The CS-OIDP measure discriminated most effectively between patients with and without hypodontia and was related to severity and upper anterior localisation of hypodontia.


Primary dental care evaluation in Brazil: an item response theory approach.
Item response theory (IRT) is a method used to design, analyze, and score tests, questionnaires, and similar instruments measuring abilities, attitudes, or other variables. The aim of this study was to assess the psychometric properties of dental care questions in the "Programa Nacional de Melhoria do Acesso e Qualidade da Atenção Básica-PMAQ-AB," Brazil, using IRT. Dentists in primary health care units in Brazil (n = 12,403) were interviewed face-to-face using a structured questionnaire. The questions were primarily dichotomous, with a no answer/do not know option. The items about dental care from a Brazilian national evaluation survey were analyzed using the IRT model (20 items). Oral health teams (OHTs) received scores that varied from the lowest performance level to the highest performance level. Scores for the Brazilian states and the Federal District were calculated to evaluate the regional distribution of OHT performance. The questions about dental care exhibited higher discrimination power for OHTs with below average performance. In general, the teams, including those with low performance, performed the actions, and procedures included in the questionnaire. Actions such as making prostheses and tracking and monitoring oral cancer cases characterized the high-performing teams. The performance of the teams distributed throughout the Brazilian states indicated that OHTs in the south and southeast performed better than OHTs in the rest of the country. Although the analyzed items are insufficient to determine the performance of OHTs, the items related to prosthesis and oral cancer tend to discriminate high-performing OHTs from other OHTs.


Stem cell growth becomes predominant while neural plate progenitor pool decreases during spinal cord elongation.
The antero-posterior dispersion of clonally related cells is a prominent feature of axis elongation in vertebrate embryos. Two major models have been proposed: (i) the intercalation of cells by convergent-extension and (ii) the sequential production of the forming axis by stem cells. The relative importance of both of these cell behaviors during the long period of elongation is poorly understood. Here, we use a combination of single cell lineage tracing in the mouse embryo, computer modeling and confocal video-microscopy of GFP labeled cells in the chick embryo to address the mechanisms involved in the antero-posterior dispersion of clones. In the mouse embryo, clones appear as clusters of labeled cells separated by intervals of non-labeled cells. The distribution of intervals between clonally related clusters correlates with a statistical model of a stem cell mode of growth only in the posterior spinal cord. A direct comparison with published data in zebrafish suggests that elongation of the anterior spinal cord involves similar intercalation processes in different vertebrate species. Time-lapse analyses of GFP labeled cells in cultured chick embryos suggest a decrease in the size of the neural progenitor pool and indicate that the dispersion of clones involves ordered changes of neighborhood relationships. We propose that a pre-existing stem zone of growth becomes predominant to form the posterior half of the axis. This temporal change in tissue-level motion is discussed in terms of the clonal and genetic continuities during axis elongation.


Palliative homecare is associated with reduced high- and low-acuity emergency department visits at the end of life: A population-based cohort study of cancer decedents.
Prior work shows that palliative homecare services reduce the subsequent need for hospitalizations and emergency services; however, no study has investigated whether this association is present for emergency department visits of high acuity or whether it only applies to low-acuity emergency department visits. To examine the association between palliative versus standard homecare nursing and the rate of high-acuity and low-acuity emergency department visits among cancer decedents during their last 6 months of life. This is a retrospective cohort study of end-of-life homecare patients in Ontario, Canada, who had confirmed cancer cause of death from 2004 to 2009. A multivariable Poisson regression analysis was implemented to examine the association between the receipt of palliative homecare nursing (vs standard homecare nursing) and the rate of high- and low-acuity emergency department visits, separately. There were 54,743 decedents who received homecare nursing in the last 6 months of life. The receipt of palliative homecare nursing decreased the rate of low-acuity emergency department visits (relative rate = 0.53, 95% confidence interval = 0.50-0.56) and was significantly associated with a larger decrease in the rate of high-acuity emergency department visits (relative rate = 0.37, 95% confidence interval = 0.35-0.38). Receiving homecare nursing with palliative intent may decrease the need for dying cancer patients to visit the emergency department, for both high and low-acuity visits, compared to receiving general homecare nursing. Policy implications include building support for additional training in palliative care to generalist homecare nurses and increasing access to palliative homecare nursing.


Allosteric modulation of rat brain nitric oxide synthase by the pterin-site enzyme inhibitor 4-aminotetrahydrobiopterin.
We investigated the functional and allosteric effects of the 4-amino analogue of tetrahydrobiopterin, (6R)-2,4-diamino- 5,6,7,8-tetrahydro-6-(L-erythro-1,2-dihydroxypropyl) pteridine (4-amino-H4biopterin) on pteridine-free rat neuronal nitric oxide synthase. In the presence of added (6R)-5,6,7,8-tetrahydro-L-erythrobiopterin (H4biopterin; 10 microM), 4-amino-H4biopterin completely inhibited the conversion of both L-arginine and NG-hydroxy-L-arginine with half-maximally effective concentrations of 1.1+/-0.09 and 1.3+/-0.09 microM, respectively. Inhibition was reversible, as shown by a time-dependent restoration of citrulline formation upon dilution of the inhibitor-treated enzyme (t1/2=3.0 min). Binding of 4-amino-H4biopterin led to a complete conversion of the haem from low-spin to high-spin state, and to the formation of stable homodimers which partially survived electrophoresis under denaturating conditions. These results show that oxidation of both L-arginine and NG-hydroxy-L-arginine is pteridine-dependent, and that the allosteric effects of H4biopterin do not fully explain the essential role of the pteridine cofactor in nitric oxide biosynthesis.


Descriptive spatial analysis of BSE in western France.
The spatial heterogeneity of Bovine Spongiform Encephalopathy (BSE) was analysed on the 84 cases confirmed in western France (WF) between August and December 2000, when both the Mandatory Reporting System and an active surveillance on cattle at risk were running. Ninety-four percent of these cases were born between June 1993 and June 1996, and we analysed the location at birth. One disease mapping and two clustering methods (Scan of Kulldorff and the method of Besag and Newell) were used. In order to attenuate the contrasts artificially created by the standard disease mapping method (over-dispersion), we estimated the Standard Incidence Ratio (SIR) with a Bayesian method (Poisson-Gamma model) allowing a smoothing of the estimators. The geographical location of interest was the "canton", that divided the total area into 526 geographical units. The background population (2.6 million cattle) was obtained from the Agricultural Census 2000. We tested the hypothesis of a homogenous spatial distribution of the BSE risk where the expected number of BSE cases per unit area was obtained by applying the overall BSE rate in WF to each "canton", standardised on the type of breed, dairy versus beef suckler. The SIR ranged from 0.80 to 2.18 and the spatial distribution of BSE cases was significantly heterogeneous. Two spatial clusters were detected with the spatial scan statistics of Kulldorff and the method of Besag and Newell (18 to 20 observed BSE-cases per cluster with a radius of 45 km) centred on the "département" of Côtes-d'Armor and Mayenne. Another cluster was detected with the method of Besag and Newell (9 observed BSE-cases) in the "département" of Finistère. The results proved that the risk of BSE is linked to the geographical location in the area of the study.


Precise Probing of Residue Roles by Post-Translational β,γ-C,N Aza-Michael Mutagenesis in Enzyme Active Sites.
Biomimicry valuably allows the understanding of the essential chemical components required to recapitulate biological function, yet direct strategies for evaluating the roles of amino acids in proteins can be limited by access to suitable, subtly-altered unnatural variants. Here we describe a strategy for dissecting the role of histidine residues in enzyme active sites using unprecedented, chemical, post-translational side-chain-β,γ C-N bond formation. Installation of dehydroalanine (as a "tag") allowed the testing of nitrogen conjugate nucleophiles in "aza-Michael"-1,4-additions (to "modify"). This allowed the creation of a regioisomer of His (iso-His, Hisiso) linked instead through its pros-Nπ atom rather than naturally linked via C4, as well as an aza-altered variant aza-Hisiso. The site-selective generation of these unnatural amino acids was successfully applied to probe the contributing roles (e.g., size, H-bonding) of His residues toward activity in the model enzymes subtilisin protease from Bacillus lentus and Mycobacterium tuberculosis pantothenate synthetase.


Machine learning ensemble models predict total charges and drivers of cost for transsphenoidal surgery for pituitary tumor.
Efficient allocation of resources in the healthcare system enables providers to care for more and needier patients. Identifying drivers of total charges for transsphenoidal surgery (TSS) for pituitary tumors, which are poorly understood, represents an opportunity for neurosurgeons to reduce waste and provide higher-quality care for their patients. In this study the authors used a large, national database to build machine learning (ML) ensembles that directly predict total charges in this patient population. They then interrogated the ensembles to identify variables that predict high charges. The authors created a training data set of 15,487 patients who underwent TSS between 2002 and 2011 and were registered in the National Inpatient Sample. Thirty-two ML algorithms were trained to predict total charges from 71 collected variables, and the most predictive algorithms combined to form an ensemble model. The model was internally and externally validated to demonstrate generalizability. Permutation importance and partial dependence analyses were performed to identify the strongest drivers of total charges. Given the overwhelming influence of length of stay (LOS), a second ensemble excluding LOS as a predictor was built to identify additional drivers of total charges. An ensemble model comprising 3 gradient boosted tree classifiers best predicted total charges (root mean square logarithmic error = 0.446; 95% CI 0.439-0.453; holdout = 0.455). LOS was by far the strongest predictor of total charges, increasing total predicted charges by approximately $5000 per day.In the absence of LOS, the strongest predictors of total charges were admission type, hospital region, race, any postoperative complication, and hospital ownership type. ML ensembles predict total charges for TSS with good fidelity. The authors identified extended LOS, nonelective admission type, non-Southern hospital region, minority race, postoperative complication, and private investor hospital ownership as drivers of total charges and potential targets for cost-lowering interventions.


Therapeutic Approach to Pulp Canal Calcification as Sequelae of Dental Avulsion.
Tooth avulsion represents the most complicated dental injury, and the classic treatment is tooth replantation. The most common sequelae are pulp canal calcification (PCC) and pulp necrosis. The presence of pulp necrosis after PCC is reported in up to 30% of the cases and is a challenge due to the difficulty of endodontic treatment. This case report describes the surgical treatment of a replanted tooth presenting PCC and periapical pathology eight years after the trauma. An endodontic surgery was performed to remove the apical granuloma, to prepare the apical root, and to seal the apical region with an endodontic cement. In a three-year follow-up, there was an absence of inflammatory signs and symptoms or apical lesion. This report shows the importance of close follow-up after dentoalveolar injuries. After the initial dental trauma and its consequences to pulpal tissues, the executed procedures allowed a favourable outcome.


Rapid radiation in spiny lobsters (Palinurus spp) as revealed by classic and ABC methods using mtDNA and microsatellite data.
Molecular tools may help to uncover closely related and still diverging species from a wide variety of taxa and provide insight into the mechanisms, pace and geography of marine speciation. There is a certain controversy on the phylogeography and speciation modes of species-groups with an Eastern Atlantic-Western Indian Ocean distribution, with previous studies suggesting that older events (Miocene) and/or more recent (Pleistocene) oceanographic processes could have influenced the phylogeny of marine taxa. The spiny lobster genus Palinurus allows for testing among speciation hypotheses, since it has a particular distribution with two groups of three species each in the Northeastern Atlantic (P. elephas, P. mauritanicus and P. charlestoni) and Southeastern Atlantic and Southwestern Indian Oceans (P. gilchristi, P. delagoae and P. barbarae). In the present study, we obtain a more complete understanding of the phylogenetic relationships among these species through a combined dataset with both nuclear and mitochondrial markers, by testing alternative hypotheses on both the mutation rate and tree topology under the recently developed approximate Bayesian computation (ABC) methods. Our analyses support a North-to-South speciation pattern in Palinurus with all the South-African species forming a monophyletic clade nested within the Northern Hemisphere species. Coalescent-based ABC methods allowed us to reject the previously proposed hypothesis of a Middle Miocene speciation event related with the closure of the Tethyan Seaway. Instead, divergence times obtained for Palinurus species using the combined mtDNA-microsatellite dataset and standard mutation rates for mtDNA agree with known glaciation-related processes occurring during the last 2 my. The Palinurus speciation pattern is a typical example of a series of rapid speciation events occurring within a group, with very short branches separating different species. Our results support the hypothesis that recent climate change-related oceanographic processes have influenced the phylogeny of marine taxa, with most Palinurus species originating during the last two million years. The present study highlights the value of new coalescent-based statistical methods such as ABC for testing different speciation hypotheses using molecular data.


Balancing torques in membrane-mediated interactions: exact results and numerical illustrations.
Torques on interfaces can be described by a divergence-free tensor which is fully encoded in the geometry. This tensor consists of two terms, one originating in the couple of the stress, the other capturing an intrinsic contribution due to curvature. In analogy to the description of forces in terms of a stress tensor, the torque on a particle can be expressed as a line integral along a contour surrounding the particle. Interactions between particles mediated by a fluid membrane are studied within this framework. In particular, torque balance places a strong constraint on the shape of the membrane. Symmetric two-particle configurations admit simple analytical expressions which are valid in the fully nonlinear regime; in particular, the problem may be solved exactly in the case of two membrane-bound parallel cylinders. This apparently simple system provides some flavor of the remarkably subtle nonlinear behavior associated with membrane-mediated interactions.


Variable and multiple expression of Protease Nexin-1 during mouse organogenesis and nervous system development.
Protease Nexin-1 (PN-1) also known as Glia-Derived Nexin (GDN) inhibits the activity of several serine proteases including thrombin, tissue (tPA)- and urokinase (uPA)-type plasminogen activators. These and other serine proteases seem to play roles in development and tissue homeostasis. To gain insight into where and when PN-1 might counteract serine protease activities in vivo, we examined its mRNA and protein expression in the mouse embryo, postnatal developing nervous system and adult tissues. These analyses revealed distinct temporal and spatial PN-1 expression patterns in developing cartilage, lung, skin, urogenital tract, and central and peripheral nervous system. In the embryonic spinal cord, PN-1 expression occurs in cells lining the neural canal that are different from the cells previously shown to express tPA. In the developing postnatal brain, PN-1 expression appears transiently in many neuronal cell populations. These findings suggest a role for PN-1 in the maturation of the central nervous system, a phase that is accompanied by the appearance of different forms of PN-1. In adults, few distinct neuronal cell populations like pyramidal cells of the layer V in the neocortex retained detectable levels of PN-1 expression. Also, mRNA and protein levels did not correspond in adult spleen and muscle tissues. The widespread and complex regulation of PN-1 expression during embryonic development and, in particular, in the early postnatal nervous system as well as in adult tissues suggests multiple roles for this serine protease inhibitor in organogenesis and tissue homeostasis.


An Unusual Mimicker of Systemic Lupus Erythematosus: A Case Report.
We present a case of a 47 year-old African American female with 15 pack-years of tobacco use and heavy alcohol use who presented with arthritis and was found to have a positive antinuclear antibodies (ANA), anti double stranded DNA antibodies (anti-dsDNA), and anti-Sjogren's syndrome-related antigen A and antigen B (anti-SSA and anti-SSB). She was subsequently found to have a lung adenocarcinoma associated with hypertrophic pulmonary osteoarthropathy (HPO). This demonstrates a case of positive antinuclear antibodies and arthritis in a patient with lung adenocarcinoma, which can be falsely diagnosed as systemic lupus erythematosus.


A method to quantify and analyze the foraging activity of honey bees: relevance to the sublethal effects induced by systemic insecticides.
The assessment of agropharmaceuticals' side effects requires more realistic simulations of field conditions than those deduced from the dose-lethality relation obtained under laboratory conditions. Because the presence of sublethal doses or concentrations may also alter the behavior of foraging insects, we attempted to devise a quantifiable and accurate protocol for evidencing various alterations in free-flying bees. Such a protocol was illustrated by testing new classes of systemic insecticides. The protocol focused on video recording to quantify the foraging activity of small colonies of honey bees confined in insect-proof tunnels. The basis of the protocol was not the colony itself but the change in each colony on a specific day and between days. First, the paradigms of attendance at a safe feeding source were established by observing 8 control colonies at different times of the season during 5 days after the necessary forager training was accomplished. Second, on three different colonies we considered the paradigms on the control day before contamination and during 4 days after the feeding source was contaminated. During the same period, one more colony was exclusively fed with safe food to serve as control. Two plant-systemic insecticides were tested at contamination levels 70 times lower than the 50% of the lethal concentration. Imidacloprid, at 6 microg/kg, clearly induced a decrease in the proportion of active bees. Fipronil, at 2 microg/kg, induced an additional decrease in attendance at the feeder. Such levels are still higher than the corresponding lowest observable effect concentration (LOEC). Our protocol, which provided intermediate conditions between field and laboratory conditions, allowed the quantification, with an enhanced level of sensitivity, of sublethal effects on foraging bees.


Stewardship in sepsis.
Sepsis is the major cause of mortality from any infectious disease worldwide. The goals of antimicrobial stewardship are to achieve optimum clinical outcomes and to ensure cost effectiveness and minimum unintended consequences, including toxic effects, selection of pathogenic organisms, and resistance. The combination of inadequate diagnostic criteria for sepsis with the extraordinary time pressure to provide broad-spectrum antimicrobial therapy is troubling from a stewardship perspective. Use of empirical therapy according to guidelines, de-escalation of therapy, switch from intravenous to oral therapy, therapeutic drug monitoring, use of a list of restricted antibiotics, and bedside consultation can lead to significant benefits for clinical outcomes, adverse events, and costs.


Acute viral hepatitis - should the current screening strategy be modified?
The epidemiology of viral hepatitis has changed. Since the introduction of safe and effective vaccines for hepatitis A and B in 1980s, the incidence of acute infections caused by these viruses has been declining in the UK. At the same time, hepatitis E virus (HEV) has been recognised as an increasingly important cause of acute hepatitis, but testing is not widely available. The aim of this study was to establish the viral causes of acute hepatitis, and use that data to modify the current diagnostic algorithm. A Cognos search was performed to collate subjects tested for HAV, HBV, HCV, HEV, EBV and CMV between June 2010 and December 2012. Information included virological result and their ALT level if done within 5 days from virological testing. From 3462 subjects with suspected acute viral hepatitis, only 25% had biochemical evidence of acute hepatitis (n=854; ALT>100IU/l). The frequency of detection of acute HEV infection (25/409) was over 31-times higher than that of HAV (6/3462), and 7-times higher than that of HBV (24/3462). Most cases of acute HAV, HEV, EBV and CMV infections presented with abnormal ALT levels. Most EBV infections were associated with lymphadenopathy (23/34); in comparison most of CMV infections were not associated with lymphadenopathy (18/22). HEV screening should be included in the initial testing panel for acute hepatitis and screening at least for HAV and HEV might be limited to those with abnormal ALT levels.


Relationship between bed sharing and breastfeeding: longitudinal, population-based analysis.
This is an investigation into the longitudinal patterns of bed sharing, the characteristics associated with those patterns, and the relationship with breastfeeding. The study used prospective, population-based data from the United Kingdom to investigate nocturnal bed sharing at 5 time points from birth to 4 years of age. Of 14 062 live births, 7447 (53%) had data available for all time points. Latent class analysis identified 4 mutually exclusive groups, broadly described as nonsharers (66%), early bed sharers (only in infancy) (13%), late bed sharers (after the first year) (15%), and constant bed sharers (throughout the 4 years) (6%). The boy/girl ratio and the proportion of families of nonwhite ethnicity were slightly higher in all 3 bed-sharing groups, compared with the non-bed-sharing group. Higher maternal educational achievement and higher social class were positively associated with early bed sharing, negatively associated with late bed sharing, and not associated with constant bed sharing. The 3 bed-sharing patterns were related significantly to breastfeeding at 12 months (P < .001), whether the families shared beds late (odds ratio: 1.72 [95% confidence interval: 1.36-2.18]), early (odds ratio: 2.36 [95% confidence interval: 1.87-2.97]), or for the whole period (odds ratio: 5.29 [95% confidence interval: 4.05-6.91]). The prevalence of breastfeeding was significantly higher among the groups that shared beds constantly or early for each of the first 15 months after birth. Advice on whether bed sharing should be discouraged needs to take into account the important relationship with breastfeeding.


Attitude toward genetic testing for cancer risk in Istanbul.
To identify attitudes toward genetic testing, and the effects of this information on decisions regarding issues such as pregnancy, abortion, and prophylactic surgery, several subsets of the Turkish population were surveyed in hospital settings. Individuals (n = 179) chosen arbitrarily from four different subsets of a Turkish population were asked to participate in a confidential 23-question survey. Survey participants were familiar with the concept of cancer being a familial disease (85.5%), and 84.7% of them expressed interest in genetic testing to determine cancer risk, 83.9% would have their fetuses tested for such cancer risk, 65.1% would terminate their pregnancies, 92.2% would have their children tested if they were determined to have an increased cancer risk, 71.9% would agree to undergo prophylactic oophorectomy or orchiectomy and 67.6% would have mastectomy/prostatectomy should there be an increased cancer risk to these organs. It appears that at least the sampled segment of a Turkish population is willing to undergo genetic testing to determine if they are at increased risk for cancer. The feasibility and acceptance of genetic testing and the influence of education and genetic counseling in the Turkish people should further be evaluated with a larger stratified sample of the population.


Development of techniques for the genetic manipulation of the gliding bacterium Cytophaga johnsonae.
Cytophaga johnsonae displays many features that make it an excellent model of bacterial gliding motility. Unfortunately, genetic analyses of C. johnsonae, or any related gliding bacteria, were not possible because of a complete lack of selectable markers, cloning vectors, transposons, and convenient methods of gene transfer. As a first step toward a molecular analysis of gliding motility of C. johnsonae, we developed these genetic techniques and tools. Common broad-host-range plasmids and transposons did not function in C. johnsonae. We identified one Bacteroides transposon, Tn4351, that could be introduced into C. johnsonae on plasmid R751 by conjugation from Escherichia coli. Tn4351 inserted in the C. johnsonae genome and conferred erythromycin resistance. Tn-4351 insertions resulted in auxotrophic mutations and motility mutations. We constructed novel plasmids and cosmids for genetic analyses of C. johnsonae. These cloning vectors are derived from a small cryptic plasmid (pCP1) that we identified in the fish pathogen Cytophaga psychrophila D12. These plasmids contain the ermF (erythromycin resistance) gene from Tn4351 and a variety of features that facilitate propagation and selection in E. coli and conjugative transfer from E. coli to C. johnsonae.


Comparing patient care seeking pathways in three models of hospital and TB programme collaboration in China.
Public hospitals in China play an important role in tuberculosis (TB) control. Three models of hospital and TB control exist in China. The dispensary model is the most common one in which a TB dispensary provides both clinical and public health care. The specialist model is similar to the former except that a specialist TB hospital is located in the same area. The specialist hospital should treat only complicated TB cases but it also treats simple cases in practice. The integrated model is a new development to integrate TB service in public hospitals. Patients were diagnosed, treated and followed up in this public hospital in this model while the TB dispensary provides public health service as case reporting and mass education. This study aims to compare patient care seeking pathways under the three models, and to provide policy recommendation for the TB control system reform in China. Six sites, two in each model, were selected across four provinces, with 293 newly treated uncomplicated TB patients being randomly selected. The majority (68%) of TB patients were diagnosed in hospitals. Patients in the integrated model presented the simplest care seeking pathways, with the least number of providers visited (2.2), shortest treatment delays (2 days) and the least medical expenditure (2729RMB/401USD). On the contrary, patients in the specialist model had the highest number of provider visits (4), longest treatment delays (23 days) and the highest medical expenditure (11626RMB/1710USD). Logistic regression suggested that patients who were hospitalised tended to have longer treatment delays and higher medical expenditure. Specialist hospital treating uncomplicated cases not using the standard regimens posed a threat to TB control. The integrated model has shortened patient treatment pathways, and reduced patient costs; therefore, it could be considered as the direction for future reform of China's TB control system.


LKB1 and AMPK family signaling: the intimate link between cell polarity and energy metabolism.
Research on the LKB1 tumor suppressor protein mutated in cancer-prone Peutz-Jeghers patients has continued at a feverish pace following exciting developments linking energy metabolism and cancer development. This review summarizes the current state of research on the LKB1 tumor suppressor. The weight of the evidence currently indicates an evolutionary conserved role for the protein in the regulation of various aspects of cellular polarity and energy metabolism. We focus on studies examining the concept that both cellular polarity and energy metabolism are regulated through the conserved LKB1-AMPK signal transduction pathway. Recent studies from a variety of model organisms have given new insight into the mechanism of polyp development and cancer formation in Peutz-Jeghers patients and the role of LKB1 mutation in sporadic tumorigenesis. Conditional LKB1 mouse models have outlined a tissue-dependent context for pathway activation and suggest that LKB1 may affect different AMPK isoforms independently. Elucidation of the molecular mechanism responsible for Peutz-Jeghers syndrome will undoubtedly reveal important insight into cancer development in the larger population.


Magnetic control of electrochemical processes at electrode surface using iron-rich graphene materials with dual functionality.
Metal-doped graphene hybrid materials demonstrate promising capabilities in catalysis and various sensing applications. There also exists great interest for on-demand control of the selectivity of many electrochemical processes. In this work, an iron-doped thermally reduced graphene oxide (Fe-TRGO) was prepared and used to investigate the possibility of a reproducible, magnetically controlled method to modulate electrochemical reactivities through a scalable method. We made use of the presence of both magnetic and electrocatalytic properties in the Fe-TRGOs to induce attraction and removal of the Fe-TRGO material onto and off the working electrode surfaces magnetically, thereby controlling the electrochemical oxidation and reduction processes. The outstanding electrochemical performance of the Fe-TRGO material was evident, with enhanced current signals and lower peak potentials observed upon magnetic activation. Reversible and reproducible cycles of activation and deactivation were obtained as the peak heights and peak potentials remained relatively consistent with no apparent carryover between every step. Both components of Fe-TRGO play an electrocatalytic role in the electrochemical sensing. In the cases of the oxygen reduction reaction and reduction of cumene hydroperoxide, the iron oxide plays the role of an electrocatalyst, while in the cases of ascorbic acid, the enhanced electroactivity originates from the high surface area of the graphene portion in the Fe-TRGO hybrid material. The feasibility of this magnetically switchable method for on-demand sensing and energy production thus brings about potential developments for future electrochemical applications.


Cu(II) potentiation of alzheimer abeta neurotoxicity. Correlation with cell-free hydrogen peroxide production and metal reduction.
Oxidative stress markers as well as high concentrations of copper are found in the vicinity of Abeta amyloid deposits in Alzheimer's disease. The neurotoxicity of Abeta in cell culture has been linked to H(2)O(2) generation by an unknown mechanism. We now report that Cu(II) markedly potentiates the neurotoxicity exhibited by Abeta in cell culture. The potentiation of toxicity is greatest for Abeta1-42 > Abeta1-40 >> mouse/rat Abeta1-40, corresponding to their relative capacities to reduce Cu(II) to Cu(I), form H(2)O(2) in cell-free assays and to exhibit amyloid pathology. The copper complex of Abeta1-42 has a highly positive formal reduction potential ( approximately +500-550 mV versus Ag/AgCl) characteristic of strongly reducing cuproproteins. These findings suggest that certain redox active metal ions may be important in exacerbating and perhaps facilitating Abeta-mediated oxidative damage in Alzheimer's disease.


The Impact of Proximal Vessel Tortuosity on the Outcomes of Chronic Total Occlusion Percutaneous Coronary Intervention: Insights From a Contemporary Multicenter Registry.
We examined the impact of proximal vessel tortuosity on the outcomes of chronic total occlusion (CTO) percutaneous coronary intervention (PCI). The baseline clinical and angiographic characteristics and procedural outcomes of 1618 consecutive CTO-PCIs performed between 2012 and 2016 at 14 United States centers in 1589 patients were reviewed. Mean patient age was 65.3 ± 10.0 years and 85% were men. Moderate/severe proximal vessel tortuosity was present in 35.7% of target lesions. Compared with non-tortuous lesions, tortuous lesions had longer length (30 mm [interquartile range, 20-50 mm] vs 28 mm [interquartile range, 16-40 mm]; P<.001), more proximal cap ambiguity (36% vs 28%; P<.01), and more frequent utilization of the retrograde approach (52% vs 37%; P<.001). Moderate/severe proximal vessel tortuosity was associated with lower technical success rates (84.1% vs 91.3%; P<.001) and procedural success rates (82.3% vs 89.9%; P<.001), but similar incidence of major cardiac adverse events (3.0% vs 2.5%; P=.59). Moderate/severe tortuosity was associated with longer procedure time and fluoroscopy time, higher air kerma radiation dose, and larger contrast volume. In a contemporary multicenter registry, moderate/severe proximal vessel tortuosity was present in approximately one-third of target CTO lesions and was associated with more frequent use of the retrograde approach and lower success rates, but similar complication rates.


Effect of mammalian lignans on the growth of prostate cancer cell lines.
Mammalian lignans, enterolactone (EL) and enterodiol (ED), have been shown to inhibit breast and colon carcinoma. To date, there have been no reports of the effect of lignans on prostatic carcinoma. We investigated the effects of ED and EL on three human prostate cancer cell lines (PC-3, DU-145 and LNCaP). Cells were treated with either 0.1% (v/v) DMSO (vehicle) or 10-100 microM of EL, ED or genistein (positive control) for 72 hours. Cell viability was measured by the propidium iodide nuclei staining fluorometric assay with each assay performed in triplicate. At 10-100 microM, EL significantly inhibited the growth of all cell lines, whereas ED only inhibited PC-3 and LNCaP cells. While EL was a more potent growth inhibitor than ED, both were less potent than genistein. The dose for 50% growth inhibition of LNCaP cells (IC50) by EL was 57 microM, whereas IC50 was 100 microM for ED, (the observed IC50 for genistein was 25 microM). ED and EL suppress the growth of prostate cancer cells, and may do so via hormonally-dependent and independent mechanisms.


MicroRNA-19a contributes to the epigenetic regulation of tissue factor in diabetes.
Diabetes mellitus is characterized by chronic vascular disorder and presents a main risk factor for cardiovascular mortality. In particular, hyperglycaemia and inflammatory cytokines induce vascular circulating tissue factor (TF) that promotes pro-thrombotic conditions in diabetes. It has recently become evident that alterations of the post-transcriptional regulation of TF via specific microRNA(miR)s, such as miR-126, contribute to the pathogenesis of diabetes and its complications. The endothelial miR-19a is involved in vascular homeostasis and atheroprotection. However, its role in diabetes-related thrombogenicity is unknown. Understanding miR-networks regulating procoagulability in diabetes may help to develop new treatment options preventing vascular complications. Plasma of 44 patients with known diabetes was assessed for the expression of miR-19a, TF protein, TF activity, and markers for vascular inflammation. High miR-19a expression was associated with reduced TF protein, TF-mediated procoagulability, and vascular inflammation based on expression of vascular adhesion molecule-1 and leukocyte count. We found plasma expression of miR-19a to strongly correlate with miR-126. miR-19a reduced the TF expression on mRNA and protein level in human microvascular endothelial cells (HMEC) as well as TF activity in human monocytes (THP-1), while anti-miR-19a increased the TF expression. Interestingly, miR-19a induced VCAM expression in HMEC. However, miR-19a and miR-126 co-transfection reduced total endothelial VCAM expression and exhibited additive inhibition of a luciferase reporter construct containing the F3 3'UTR. While both miRs have differential functions on endothelial VCAM expression, miR-19a and miR-126 cooperate to exhibit anti-thrombotic properties via regulating vascular TF expression. Modulating the post-transcriptional control of TF in diabetes may provide a future anti-thrombotic and anti-inflammatory therapy.


Gold immunoblot analysis of IgM-specific antibody in the diagnosis of human leptospirosis.
An immunoblot for the detection of leptospirosis was developed in our laboratory. Antigen prepared from Leptospira interrogans serovar bataviae was dotted onto nitrocellulose paper and blocked with skim milk. Test and control sera diluted 1:20 were applied to the dot, incubated, and washed. Anti-human IgM colloidal gold conjugate was added and the dots were washed. A positive reaction was shown by the development of a pink dot against a white background. The test was performed on 62 sera that tested positive for leptospirosis by a microagglutination (MA) test, on 40 sera that were positive by an indirect hemagglutination (IHA) test, and on sera from forty healthy blood donors. Four sera from the blood donors showed a faint pink dot, but the remainder showed a colorless reaction. All 62 sera that tested positive by MA were positive by this new test, while 95% of the 20 sera that tested positive by IHA were positive. Tests for IgG antibody were performed on 20 sera positive by MA using protein A-colloidal gold conjugate, and all showed weak reactivity. The results confirmed previous findings that most antibodies present in leptospirosis patients are of the IgM type. The ELISA takes three hours to perform, but the gold immunoblot can be completed in 30 min. In addition, the test blot can be kept as a permanent record, and is a significant improvement over existing tests.


S3QL: a distributed domain specific language for controlled semantic integration of life sciences data.
The value and usefulness of data increases when it is explicitly interlinked with related data. This is the core principle of Linked Data. For life sciences researchers, harnessing the power of Linked Data to improve biological discovery is still challenged by a need to keep pace with rapidly evolving domains and requirements for collaboration and control as well as with the reference semantic web ontologies and standards. Knowledge organization systems (KOSs) can provide an abstraction for publishing biological discoveries as Linked Data without complicating transactions with contextual minutia such as provenance and access control.We have previously described the Simple Sloppy Semantic Database (S3DB) as an efficient model for creating knowledge organization systems using Linked Data best practices with explicit distinction between domain and instantiation and support for a permission control mechanism that automatically migrates between the two. In this report we present a domain specific language, the S3DB query language (S3QL), to operate on its underlying core model and facilitate management of Linked Data. Reflecting the data driven nature of our approach, S3QL has been implemented as an application programming interface for S3DB systems hosting biomedical data, and its syntax was subsequently generalized beyond the S3DB core model. This achievement is illustrated with the assembly of an S3QL query to manage entities from the Simple Knowledge Organization System. The illustrative use cases include gastrointestinal clinical trials, genomic characterization of cancer by The Cancer Genome Atlas (TCGA) and molecular epidemiology of infectious diseases. S3QL was found to provide a convenient mechanism to represent context for interoperation between public and private datasets hosted at biomedical research institutions and linked data formalisms.


Radical cations of phenyl silatrane.
Electrochemical oxidation of phenylsilatrane (1) in CH3CN/0.1 M Bu4NPF6 has been studied by voltammetry, UV-Vis and EPR-coupled spectroelectrochemistry supported by DFT calculations. One-electron withdrawal from the HOMO of 1, formed with a predominant contribution of the atrane N atom to the 3c-4e system, results in a short lived radical cation, in which the atrane nitrogen atom is almost planar and carries most of the spin density showing strong coupling with the protons of the axially directed C-H bonds of the three adjacent α-methylene groups (g = 2.0037, aαHax = 37.93 G, aαHlat = 0.23 G and aβH = 1.8 G). EPR spectroscopy and DFT calculations attest that the unpaired electron in the radical cation does not reside at the Si atom.


Endogenous Sensory Discrimination and Selection by a Fast Brain Switch for a High Transfer Rate Brain-Computer Interface.
In this study, we present a novel multi-class brain-computer interface (BCI) for communication and control. In this system, the information processing is shared by the algorithm (computer) and the user (human). Specifically, an electro-tactile cycle was presented to the user, providing the choice (class) by delivering timely sensory input. The user discriminated these choices by his/her endogenous sensory ability and selected the desired choice with an intuitive motor task. This selection was detected by a fast brain switch based on real-time detection of movement-related cortical potentials from scalp EEG. We demonstrated the feasibility of such a system with a four-class BCI, yielding a true positive rate of ∼ 80% and ∼ 70%, and an information transfer rate of ∼ 7 bits/min and ∼ 5 bits/min, for the movement and imagination selection command, respectively. Furthermore, when the system was extended to eight classes, the throughput of the system was improved, demonstrating the capability of accommodating a large number of classes. Combining the endogenous sensory discrimination with the fast brain switch, the proposed system could be an effective, multi-class, gaze-independent BCI system for communication and control applications.


Simulated diabetic ketoacidosis therapy in vitro elicits brain cell swelling via sodium-hydrogen exchange and anion transport.
A common complication of type 1 diabetes mellitus is diabetic ketoacidosis (DKA), a state of severe insulin deficiency. A potentially harmful consequence of DKA therapy in children is cerebral edema (DKA-CE); however, the mechanisms of therapy-induced DKA-CE are unknown. Our aims were to identify the DKA treatment factors and membrane mechanisms that might contribute specifically to brain cell swelling. To this end, DKA was induced in juvenile mice with the administration of the pancreatic toxins streptozocin and alloxan. Brain slices were prepared and exposed to DKA-like conditions in vitro. Cell volume changes were imaged in response to simulated DKA therapy. Our experiments showed that cell swelling was elicited with isolated DKA treatment components, including alkalinization, insulin/alkalinization, and rapid reductions in osmolality. Methyl-isobutyl-amiloride, a nonselective inhibitor of sodium-hydrogen exchangers (NHEs), reduced cell swelling in brain slices elicited with simulated DKA therapy (in vitro) and decreased brain water content in juvenile DKA mice administered insulin and rehydration therapy (in vivo). Specific pharmacological inhibition of the NHE1 isoform with cariporide also inhibited cell swelling, but only in the presence of the anion transport (AT) inhibitor 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid. DKA did not alter brain NHE1 isoform expression, suggesting that the cell swelling attributed to the NHE1 was activity dependent. In conclusion, our data raise the possibility that brain cell swelling can be elicited by DKA treatment factors and that it is mediated by NHEs and/or coactivation of NHE1 and AT.


The effect of hypointense white matter lesions on automated gray matter segmentation in multiple sclerosis.
Previous imaging studies assessing the relationship between white matter (WM) damage and matter (GM) atrophy have raised the concern that Multiple Sclerosis (MS) WM lesions may affect measures of GM volume by inducing voxel misclassification during intensity-based tissue segmentation. Here, we quantified this misclassification error in simulated and real MS brains using a lesion-filling method. Using this method, we also corrected GM measures in patients before comparing them with controls in order to assess the impact of this lesion-induced misclassification error in clinical studies. We found that higher WM lesion volumes artificially reduced total GM volumes. In patients, this effect was about 72% of that predicted by simulation. Misclassified voxels were located at the GM/WM border and could be distant from lesions. Volume of individual deep gray matter (DGM) structures generally decreased with higher lesion volumes, consistent with results from total GM. While preserving differences in GM volumes between patients and controls, lesion-filling correction revealed more lateralised DGM shape changes in patients, which were not evident with the original images. Our results confirm that WM lesions can influence MRI measures of GM volume and shape in MS patients through their effect on intensity-based GM segmentation. The greater effect of lesions at increasing levels of damage supports the use of lesion-filling to correct for this problem and improve the interpretability of the results. Volumetric or morphometric imaging studies, where lesion amount and characteristics may vary between groups of patients or change over time, may especially benefit from this correction.


Methods of choice in completed suicides: gender differences and review of literature.
Suicide is an important public health hazard worldwide. A 4-year retrospective study from January 2000 to December 2003 was conducted to identify the favored methods in realized suicides among males and females in the west coastal region of India. During the study period, a total of 539 cases of suicidal deaths were autopsied. Males were predominantly affected (male: female-1.9:1). The age of the victims ranged from 13 to 90 years in males (mean = 40.1 years, median = 37.0 years) and 15 to 85 years in females (mean = 36.6 years, median = 32.0 years). Most favored method of suicide amongst males and females was hanging (36.9%, n = 199) followed by poisoning (34.7%, n = 187). Male dominance was apparent for each method of suicide except for self-immolation. Males were relatively more likely to use hanging and poisoning while females were more likely to prefer drowning and self-immolation as methods of suicide. Relatively younger females (mean = 33.0 years, median = 32.0 years) preferred hanging as a method of suicide when compared to males (mean = 42.4 years, median = 40.0 years). Among females, significantly younger females resorted to hanging when compared to older females who preferred drowning. On investigating the various theories proposed for choice of suicide methods in males and females in different regions we conclude that preference of method of suicide in men and women is complexly determined. In this region, availability, accessibility, popularity, and socioacceptability seem to be the major determinants in the choice of methods among males and females rather than violence associated and lethality of the method. Females were as likely to use lethal and violent methods as males in this region.


Chromatin remodeling as a mechanism for circadian prolactin transcription: rhythmic NONO and SFPQ recruitment to HLTF.
Most clock-controlled genes (CCGs) lack the specific E-box response element necessary for direct circadian regulation. This is the case for the prolactin (Prl) gene, the expression of which oscillates in individual lactotrope pituitary cells. To characterize the processes underlying this oscillation, we used a lactotrope cell line (GH4C1 cells). In these cells, Prl gene expression fluctuated significantly during 24 h (P=0.0418). Circadian Prl transcription depended on an interaction between the pituitary-specific transcription factor, PIT-1, and the helicase-like transcription factor (HLTF), a SWI/SNF chromatin remodeler, shown here to bind the Prl promoter on an E-box that differs from the specific E-box preferentially bound by clock proteins. Circadian Prl transcription was further accompanied by marked daily chromatin transitions. While neither HLTF nor PIT-1 was rhythmically expressed, NONO and SFPQ, identified as HLTF-associated proteins by mass spectrometry, displayed a circadian pattern and bound rhythmically to the Prl promoter. Furthermore, NONO and SFPQ were functionally involved in circadian Prl transcription since overexpression of both proteins greatly reduced Prl promoter activity (P<0.001) and disrupted its circadian pattern. A mechanism involving a rhythm in paraspeckle protein recruitment is proposed to explain how the core oscillator can generate a circadian pattern of CCGs lacking the specific E-box response element.


Biosynthesis of selenoproteins in cultured bovine mammary cells.
The biosynthesis of selenoproteins was studied in relation to milk formation and mammary cell biology by incubating the bovine mammary cell line MAC-T with ((75)Se)selenite. Intracellular proteins and proteins secreted into the cell culture medium were separated by 2D electrophoresis, the selenoproteins were detected by autoradiography, and the proteins were identified by MALDI-TOF. Approximately 35 (75)Se-containing spots were found in the cell proteins from MAC-T cells. Among them, one-third showed high intensity. The strongest spot was identified as glutathione peroxidase 1. About 20 spots were observed in protein precipitated from cell culture medium, one-third of them being distinctly visible. In an attempt to study a perturbation of the system, the effect of retinoic acid (RA) on the formation of selenoproteins was investigated. The concentration of (75)Se in total cell protein was reduced by about 35% in cells cultured with RA compared with control cells, while the opposite effect was observed in protein precipitated from cell culture medium, which contained 60% more (75)Se in RA-treated samples than in controls. There were also indications that RA might affect different selenoproteins in different ways. The methods described provide a promising approach for further studies of the regulation of selenoprotein formation in the mammary gland.


Racial comparison of outcomes of male Department of Veterans Affairs patients with lung and colon cancer.
A study was performed to distinguish between genetic vs socioeconomic factors or artifacts inherent in retrospective studies to discover explanations for the apparent shorter survival of blacks than whites with lung and colon cancer. Detailed biological and socioeconomic data were collected prospectively from a large cohort of patients with advanced lung and colon cancer who had not previously received chemotherapy and who consented to be entered in the clinical trial. Twelve medical centers within the Department of Veterans Affairs hospital system (Veterans Affairs Cooperative Study 188) entered patients between May 1981 and May 1986. These patients were evaluated and treated in a uniform manner within this hospital system and followed up to death. Outcome measures included time to tumor progression, tumor response, and survival. A total of 719 patients were entered in the study, 704 of whom were either black or white men. Blacks were found to have a significantly lower socioeconomic status than whites as measured by marital and educational status, occupation, income, housing, and number of individuals residing in household. Blacks had a significantly lower frequency of prior tumor resection and a significantly increased frequency of mediastinal lymph node involvement and of metastasis than whites. Trends existed toward a shorter time interval from original diagnosis to entry and a lower frequency of prior radiation therapy among blacks in comparison with whites at the time of entry to the study. However, no differences in intensity of treatment or follow-up, time to progression, response to treatment, or overall survival (measured from entry to the study to death) were observed for blacks vs whites. Lung and colon cancer are not necessarily more aggressive diseases in blacks than in whites. Based on a comparison of these data with those reported from other practice settings, we postulate that lung and colon cancer outcomes may be similar among black and white patients who receive equal access to comparable medical care in spite of socioeconomic differences. Findings suggest that future studies might focus profitably on racial factors that may exist at the time of patient entry to the health care system.


How cooperative are protein folding and unfolding transitions?
A thermodynamically and kinetically simple picture of protein folding envisages only two states, native (N) and unfolded (U), separated by a single activation free energy barrier, and interconverting by cooperative two-state transitions. The folding/unfolding transitions of many proteins occur, however, in multiple discrete steps associated with the formation of intermediates, which is indicative of reduced cooperativity. Furthermore, much advancement in experimental and computational approaches has demonstrated entirely non-cooperative (gradual) transitions via a continuum of states and a multitude of small energetic barriers between the N and U states of some proteins. These findings have been instrumental towards providing a structural rationale for cooperative versus noncooperative transitions, based on the coupling between interaction networks in proteins. The cooperativity inherent in a folding/unfolding reaction appears to be context dependent, and can be tuned via experimental conditions which change the stabilities of N and U. The evolution of cooperativity in protein folding transitions is linked closely to the evolution of function as well as the aggregation propensity of the protein. A large activation energy barrier in a fully cooperative transition can provide the kinetic control required to prevent the accumulation of partially unfolded forms, which may promote aggregation. Nevertheless, increasing evidence for barrier-less "downhill" folding, as well as for continuous "uphill" unfolding transitions, indicate that gradual non-cooperative processes may be ubiquitous features on the free energy landscape of protein folding.


Evaluation the relationship of left ventricular global longitudinal strain and laboratory parameters in discharged patients with COVID-19: a follow-up study.
The novel coronavirus infection (COVID-19) disease has spread rapidly and posed a great threat to global public health. The laboratory parameters and clinical outcomes of the disease in discharged patients remain unknown. In this study, we aimed to investigate the laboratory and echocardiographic findings of patients with COVID-19 after discharge and the relation between left ventricular global longitudinal strain (LVGLS) and inflammatory parameters in discharged patients. A total of 75 patients recovering from COVID-19 as the study group were prospectively recruited from the COVID-19 outpatient clinic for their follow-up visits at a median 6 months after discharge. Patients were classified into groups according to pneumonia severity and impairment in LVGLS. Laboratory findings of patients both at admission and after discharge were evaluated and the relation with pneumonia severity at admission and LVGLS after discharge were analyzed. Serum ferritin, lactate dehydrogenase (LDH) and prohormone B-type natriuretic peptide (pro-BNP) levels after discharge were significantly higher in the study group than the control group (n = 44). Ferritin was found to be related to pneumonia severity. Serum ferritin and LDH values after discharge were significantly higher in patients with impaired LVGLS than those with preserved. There was a significant correlation between LVGLS, serum ferritin and LDH values after discharge (r = -0.252, p = 0.012; r = -0.268, p = 0.005, respectively). Clinicians should pay close attention to the serum ferritin and LDH levels in discharged patients for predicting the severity of COVID-19 disease and early identification of subclinical left ventricular myocardial dysfunction.


Protein energy malnutrition in India: the plight of our under five children.
Protein energy malnutrition (PEM) is a major public health problem in India. This affects the child at the most crucial period of time of development, which can lead to permanent impairment in later life. PEM is measured in terms of underweight (low weight for age), stunting (low height for age) and wasting (low weight for height). The prevalence of stunting among under five is 48% and wasting is 19.8% and with an underweight prevalence of 42.5%, it is the highest in the world. Undernutrition predisposes the child to infection and complements its effect in contributing to child mortality. Lalonde model (1974) is used to look into the various determinants of PEM in under five children and its interrelation in causation of PEM. The determinants of PEM are broadly classified under four distinct categories: Environmental factors including the physical and social environment, behavioral factors, health-care service related and biological factors. The socio-cultural factors play an important role wherein, it affects the attitude of the care giver in feeding and care practices. Faulty feeding practice in addition to poor nutritional status of the mother further worsens the situation. The vicious cycle of poor nutritional status of the mother leading to low birth weight child further exposes the child to susceptibility to infections which aggravates the situation. However, it is seen that percapita income of the family did not have much bearing on the poor nutritional status of the child rather lack of proper health-care services adversely contributed to poor nutritional status of the child. PEM is a critical problem with many determinants playing a role in causing this vicious cycle of undernutrition. With almost half of under five children undernourished in India, the Millennium Development Goal (MDG) of halving the prevalence of underweight by 2015 seems a distant dream.


Circadian Rest-Activity Pattern Changes in Aging and Preclinical Alzheimer Disease.
Circadian rhythm disturbances occur in symptomatic Alzheimer disease (AD) and have been hypothesized to contribute to disease pathogenesis. However, it is unknown whether circadian changes occur during the presymptomatic phase of the disease. To examine the associations between circadian function, aging, and preclinical AD pathology in cognitively normal adults. This cross-sectional study was conducted using community volunteers from the Knight Alzheimer's Disease Research Center at Washington University in St Louis. Cognitively normal participants (n = 205) underwent 7 to 14 days of actigraphy in their home environment between 2010 and 2012, in addition to clinical assessment, amyloid imaging with Pittsburgh Compound B (PiB), and cerebrospinal fluid biomarker collection. Data collected from 3 years before to 6 months after actigraphy were included. Sixteen participants were excluded owing to incomplete data collection. Circadian rhythm analysis was performed on actigraphy data using 3 methods: cosinor, nonparametric, and empirical mode decomposition. Preclinical AD was assessed by longitudinal clinical assessment, amyloid imaging with PiB, and cerebrospinal fluid biomarker collection. Data from 189 participants were included in the analyses. The mean (SD) age was 66.6 (8.3) years, and 121 participants (64%) were women. Older age (β = .247; P = .003) and male sex (β = .170; P = .04), in the absence of amyloid pathology, were associated with a significant increase in intradaily variability, a nonparametric measure of rest-activity rhythm fragmentation, as well as decreased amplitude by several measures. After correction for age and sex, the presence of preclinical amyloid plaque pathology, assessed by positive PiB imaging (mean [SD], 0.804 [0.187] for PiB negative vs 0.875 [0.178] for PiB positive; P = .05) or increasing cerebrospinal fluid phosphorylated-tau to amyloid β 42 ratio (β = .231; P = .008), was associated with increased intradaily variability, indicating rest-activity rhythm fragmentation. Preclinical AD is associated with rest-activity rhythm fragmentation, independent of age or sex. Aging was also associated with circadian dysfunction independently of preclinical AD pathology, particularly in men. The presence of circadian rhythm abnormalities in the preclinical phase of AD suggests that circadian dysfunction could contribute to early disease pathogenesis or serve as a biomarker of preclinical disease.


The intravascular oxygenator (IVOX): preliminary results of a new means of performing extrapulmonary gas exchange.
Conventional management of adult respiratory distress syndrome (ARDS) with high minute ventilation, positive end-expiratory pressure (PEEP), and increased fractional inspired oxygen (FIO2) concentrations may worsen pulmonary injury. The intravascular oxygenator (IVOX) is a device made up of several hundred gas permeable hollow fibers that are inserted into the vena cava by femoral venous cutdown. Flow of gas through each fiber adds O2 and removes CO2 from the bloodstream. The purpose of this study was to determine if the IVOX significantly reduces the level of mechanical ventilatory support in ARDS patients. The IVOX was inserted in nine patients, and aborted in one because of technical complications. The IVOX increased PaO2 and reduced PaCO2, but the quantity of gas transfer was not sufficient to allow a reduction in PEEP, FIO2, or minute ventilation. Insertion of the IVOX decreased cardiac index and systemic oxygen delivery despite maximum fluid and inotropic support. Mortality was 80%. Although some gas exchange occurs, the current device does not allow a significant reduction in the level of mechanical ventilatory support and adversely affects systemic oxygen transport.


Having to Be the One: Mothers Providing Home Care to Infants With Complex Cardiac Needs.
Early diagnosis of complex congenital heart disease and advances in surgical interventions have resulted in remarkable improvements in prognoses and hospital survival. Although studies have provided insight into children's experiences with complex congenital heart disease after hospitalization, few have addressed parents' experiences providing care for infants with complex congenital heart disease who are discharged home with complex care needs after surgical palliation. To describe the perceptions and lived experiences of mothers of infants who were discharged from the hospital after surgery for complex congenital heart disease but were then readmitted to the hospital. Data collection and analysis for this pilot study were guided by grounded theory. From February through October 2017, interviews were conducted with 10 mothers about their experiences caring for their infants at home after surgery for complex congenital heart disease. Analyses led to development of 1 category, "having to be the one," which had 3 properties: having no choice but to provide complex care at home, handling unexpected roles, and grappling with the possibility of death. The category of "having to be the one" highlighted mothers' experiences providing medicalized care at home to their infants after complex cardiac surgery while managing other responsibilities, such as employment, busy households, and parenting other school-age children. The role of the caregiver is vital but demanding. Mothers' caregiving at home may be enhanced by nursing interventions such as routine screening for infant distress plus assessment for alterations in family coping or relational challenges that threaten family function.


Intrinsic Dynamics and Causality in Correlated Motions Unraveled in Two Distinct Inactive States of Human β2-Adrenergic Receptor.
The alternative inactive state of the human β2-adrenergic receptor originally exposed in molecular dynamics simulations was investigated using various analysis tools to evaluate causality between correlated residue-pair fluctuations and suggest allosteric communication pathways. A major conformational shift observed in the third intracellular loop (ICL3) displayed a novel inactive state, featuring an inaccessible G protein binding site blocked by ICL3 and an expanded orthosteric ligand binding site. Residue-based mean-square fluctuation and stiffness calculations revealed a significant mobility decrease in ICL3, which induced a mobility increase in the remaining loop regions. This indicates conformational entropy loss in one mobile region being compensated by residual intermolecular motions in other mobile regions. Moreover, the extent of significantly correlated motions decreased, and correlations that once existed between transmembrane helices shifted toward regions with increased mobility. Conditional time-delayed cross-correlation analysis identified distinct driver-follower relationship profiles. Prior to its packing, freely moving ICL3 was markedly driven by transmembrane helix-8 whereas once packed, ICL3 controlled future fluctuations of nearby helices. Moreover, two transmembrane helices, (H5 and H6), started to control future fluctuations of a remote site, the extracellular loop, ECL2. This clearly suggests that allosteric coupling between extra- and intracellular parts intensified, in agreement with the receptor's well recognized feature, which is the inverse proportionality between activity and the degree of coupling.


Hospitalization for respiratory syncytial virus in the paediatric population in Spain.
The aim of this population-based retrospective study was to determine the incidence of hospitalization for community-acquired, laboratory-confirmed respiratory syncytial virus (RSV) infection in an unselected paediatric population from southern Europe. The study was performed in an area with 15,700 children aged less than 5 years attended by a single hospital. The presence of RSV in nasopharyngeal aspirates from children with acute respiratory infection treated in the hospital was investigated in four seasons (July 1996-June 2000). A total of 390 episodes of hospitalization for RSV infection were detected and 83.3% of the children were aged less than 1 year old. The annual hospitalization rate was 37/1000 for infants aged less than 6 months and 25/1000 for those aged less than 1 year. During the study period, 2.5% of the infants younger than 1 year and approximately 5% of those younger than 3 months were hospitalized for RSV infection. The mean length of hospital stay was 5.9 days. Seven per cent of the patients required admission to the intensive care unit and more than half of these children were aged less than 1 month. In Spain, community-acquired RSV infection is a highly frequent cause of hospitalization in young children, especially in those aged less than 1 year. Prevention of RSV infection, through the development of vaccines and/or other strategies, should be a public health priority.


Fine-Needle Aspiration Biopsy of Liver Lesions Yields Higher Tumor Fraction for Molecular Studies: A Direct Comparison With Concurrent Core Needle Biopsy.
This retrospective study evaluated and compared the diagnostic accuracy and suitability of tissue specimens for advanced molecular diagnostic testing obtained via 2 different techniques for percutaneous biopsy of primary and metastatic liver tumors. Samples from 137 patients with liver masses who underwent concurrent fine-needle aspiration biopsy with cell block (FNAB-CB) and core needle biopsy (CNB) at 2 hospitals were assessed for diagnostic accuracy, tumor fraction, and tumor cellularity. A subset of FNAB-CBs, that were deemed to have less or equal tumor cellularity compared with CNBs, had level sections performed and were reassessed for tumor cellularity. Diagnostic accuracy was 96% for FNAB and 93% for CNB (P=.267). In FNAB-CBs, tumor fraction was significantly higher than in CNB samples (67% vs 36%; P<.0001), whereas nontumor components were significantly lower (stromal component, 7% vs 29%; P<.0001; background benign hepatocytes, 25% vs 36%; P=.003). Additionally, in 44% of cases, FNAB-CB tumor cellularity was equal to or greater than that of the concurrent CNB. In the current age of personalized medicine, a minimally invasive, safe approach to obtaining adequate tissue for myriad molecular testing is paramount. We have shown that FNAB sampling is diagnostically accurate and produces higher tumor fractions than CNB. Thus, FNAB should be strongly considered as an initial sampling modality, especially for patients in whom molecular tests will determine management.


Improving Care after Acute Kidney Injury: A Prospective Time Series Study.
Acute kidney injury (AKI) complicates 15-20% of hospitalizations, and AKI survivors are at increased risk of chronic kidney disease and death. However, less than 20% of patients see a nephrologist within 3 months of discharge, even though a nephrologist visit within 90 days of discharge is associated with enhanced survival. To address this, we established an AKI Follow-Up Clinic and characterized the patterns of care delivered. We conducted a prospective time series study. All hospitalized patients who developed Kidney Disease Improving Global Outcomes (KDIGO) stage 2 or 3 AKI were eligible. The pre-intervention period consisted of electronic reminders to the nephrology consults and cardiovascular surgery services to refer to the AKI Follow-Up Clinic. In the post-intervention period, eligible patients were automatically scheduled into the AKI Follow-Up Clinic at discharge. The primary outcome was the percentage of KDIGO stages 2-3 AKI survivors assessed by a nephrologist within 30 days of discharge. In the pre-intervention period, 8 of 46 patients (17%) were seen by a nephrologist within 30 days after discharge, and no additional patients were seen for 90 days. In the post-intervention period, 17 of 69 patients (25%) were seen by a nephrologist within 30 days after discharge (p = 0.36), with an additional 30 patients seen in 90 days (47 of 69, 68%, p < 0.001). The mean serum creatinine was 99 (SD 35) µmol/l prior to hospitalization and 133 (58) µmol/l at 3 months. Fifty-five of 79 patients (70%) received at least 1 medical intervention at their first AKI Follow-Up Clinic visit. An AKI Follow-Up Clinic with an automatic referral process increased the proportion of patients seen at 90 days, but not 30 days post discharge. Being seen in the AKI Follow-Up Clinic was associated with interventions in most patients. Future research is needed to evaluate the effect of the AKI Follow-Up Clinic on patient-centered outcomes, but physicians should be aware that AKI survivors may benefit from close outpatient follow-up and a multipronged approach to care similarly for other high-risk populations.