Inherent dynamics of head domain correlates with ATP-recognition of P2X4 receptors: insights gained from molecular simulations.
P2X receptors are ATP-gated ion channels involved in many physiological functions, and determination of ATP-recognition (AR) of P2X receptors will promote the development of new therapeutic agents for pain, inflammation, bladder dysfunction and osteoporosis. Recent crystal structures of the zebrafish P2X4 (zfP2X4) receptor reveal a large ATP-binding pocket (ABP) located at the subunit interface of zfP2X4 receptors, which is occupied by a conspicuous cluster of basic residues to recognize triphosphate moiety of ATP. Using the engineered affinity labeling and molecular modeling, at least three sites (S1, S2 and S3) within ABP have been identified that are able to recognize the adenine ring of ATP, implying the existence of at least three distinct AR modes in ABP. The open crystal structure of zfP2X4 confirms one of three AR modes (named AR1), in which the adenine ring of ATP is buried into site S1 while the triphosphate moiety interacts with clustered basic residues. Why architecture of ABP favors AR1 not the other two AR modes still remains unexplored. Here, we examine the potential role of inherent dynamics of head domain, a domain involved in ABP formation, in AR determinant of P2X4 receptors. In silico docking and binding free energy calculation revealed comparable characters of three distinct AR modes. Inherent dynamics of head domain, especially the downward motion favors the preference of ABP for AR1 rather than AR2 and AR3. Along with the downward motion of head domain, the closing movement of loop139-146 and loop169-183, and structural rearrangements of K70, K72, R298 and R143 enabled ABP to discriminate AR1 from other AR modes. Our observations suggest the essential role of head domain dynamics in determining AR of P2X4 receptors, allowing evaluation of new strategies aimed at developing specific blockers/allosteric modulators by preventing the dynamics of head domain associated with both AR and channel activation of P2X4 receptors.


Mathematical model developed for environmental samples: prediction of GC/MS dioxin TEQ from XDS-CALUX bioassay data.
Remediation of hazardous waste sites requires efficient and cost-effective methods to assess the extent of contamination by toxic substances including dioxin-like chemicals. Traditionally, dioxin-like contamination has been assessed by gas chromatography/high-resolution mass spectrometry (GC/MS) analysis for specific polychlorinated dibenzo-p-dioxins, dibenzofurans, and biphenyl congeners. Toxic equivalency factors for these congeners are then used to estimate the overall dioxin toxic equivalency (TEQ) of complex mixtures found in samples. The XDS-CALUX bioassay estimates contamination by dioxin-like chemicals in a sample extract by measuring expression of a sensitive reporter gene in genetically engineered cells. The output of the XDS-CALUX assay is a CALUX-TEQ value, calibrated based on TCDD standards. Soil samples taken from a variety of hazardous waste sites were measured using the XDS-CALUX bioassay and GC/MS. TEQ and CALUX-TEQ from these methods were compared, and a mathematical model was developed describing the relationship between these two data sets: log(TEQ) = 0.654 x log(CALUX-TEQ) + 0.058-(log(CALUX-TEQ))2. Applying this equation to these samples showed that predicted and GC/MS measured TEQ values strongly correlate (R2 = 0.876) and that TEQ values predicted from CALUX-TEQ were on average nearly identical to the GC/MS-TEQ. The ability of XDS-CALUX bioassay data to predict GC/MS-derived TEQ data should make this procedure useful in risk assessment and management decisions.


Effect of the antiplatelet drug dilazep dihydrochloride on urinary podocytes in patients in the early stage of diabetic nephropathy.
To determine whether the antiplatelet drug dilazep dihydrochloride affects the number of urinary podocytes in diabetic patients with microalbuminuria. Fifty patients with type 2 diabetes and microalbuminuria (30 men and 20 women, mean age 48.6 years) and 30 age-matched control subjects (18 men and 12 women, mean age 49.2 years) were included in the study. No patients showed serum creatinine levels in excess of 2.0 mg/dl. Urinary podocytes were examined by immunofluorescence microscopy with monoclonal antibodies against podocalyxin. Urinary podocytes were detected in 18 of the 50 microalbuminuric diabetic patients (mean, 1.3 cells/ml). Urinary podocytes were not detected in the remaining 32 patients or in the 30 healthy control subjects. Diabetic patients positive for urinary podocytes were divided into 2 treatment groups: a dilazep dihydrochloride treatment group (300 mg/day; n = 9, group A) and a placebo group (n = 9, group B). Treatments were continued for 6 months. In group A, microalbuminuria decreased significantly from 146 +/- 42 to 86 +/- 28 microg/min (P < 0.01) and urinary podocytes also decreased from 1.3 +/- 0.8 to 0.4 +/- 0.2 cells/ml (P < 0.01). However, in group B, microalbuminuria and urinary podocytes changed little over the study period. Podocyte injury may occur in patients with early diabetic nephropathy, and dilazep dihydrochloride may be useful for preventing glomerular injury.


Song variation in an avian ring species.
Divergence of mating signals can occur rapidly and be of prime importance in causing reproductive isolation and speciation. A ring species, in which two reproductively isolated taxa are connected by a chain of intergrading populations, provides a rare opportunity to use spatial variation to reconstruct the history of divergence. I use geographic variation in the song of a likely ring species, the greenish warbler (Phylloscopus trochiloides) to reconstruct the microevolutionary steps that occurred during divergence of a trait that is often important in speciation in birds. Populations of a western Siberian (P. t. viridanus) and an eastern Siberian (P. t. plumbeitarsus) form of the greenish warbler meet, but do not interbreed in central Siberia; these forms are connected by a chain of interbreeding populations extending in a ring to the south around the treeless Tibetan Plateau. I show that: (1) song structure differs greatly between the two Siberian forms, which share the same habitat; (2) song structure changes gradually around the ring; (3) singing behavior is relatively simple in the Himalayas, but becomes increasingly complex to the north, both to the west and east of the Tibetan Plateau; and (4) song varies along independent axes of complexity in the western and eastern south-north clines. By comparing geographic variation in singing behavior and ecological variables, I distinguish among possible causes of song divergence, including selection based on the acoustic environment, stochastic effects of sexual selection, and selection for species recognition. I suggest that parallel south-to-north ecological gradients have caused a greater intensity of sexual selection on song in northern populations and that the stochastic effects of sexual selection have led to divergence in song structure.


Discrimination Based on Personal Responsibility: Luck Egalitarianism and Healthcare Priority Setting.
Luck egalitarianism is a responsibility-sensitive theory of distributive justice. Its application to health and healthcare is controversial. This article addresses a novel critique of luck egalitarianism, namely, that it wrongfully discriminates against those responsible for their health disadvantage when allocating scarce healthcare resources. The philosophical literature about discrimination offers two primary reasons for what makes discrimination wrong (when it is): harm and disrespect. These two approaches are employed to analyze whether luck egalitarian healthcare prioritization should be considered wrongful discrimination. Regarding harm, it is very plausible to consider the policies harmful but much less reasonable to consider those responsible for their health disadvantages a socially salient group. Drawing on the disrespect literature, where social salience is typically not required for something to be discrimination, the policies are a form of discrimination. They are, however, not disrespectful. The upshot of this first assessment of the discrimination objection to luck egalitarianism in health is, thus, that it fails.


A synthetic uric acid analog accelerates cutaneous wound healing in mice.
Wound healing is a complex process involving intrinsic dermal and epidermal cells, and infiltrating macrophages and leukocytes. Excessive oxidative stress and associated inflammatory processes can impair wound healing, and antioxidants have been reported to improve wound healing in animal models and human subjects. Uric acid (UA) is an efficient free radical scavenger, but has a very low solubility and poor tissue penetrability. We recently developed novel UA analogs with increased solubility and excellent free radical-scavenging properties and demonstrated their ability to protect neural cells against oxidative damage. Here we show that the uric acid analog (6, 8 dithio-UA, but not equimolar concentrations of UA or 1, 7 dimethyl-UA) modified the behaviors of cultured vascular endothelial cells, keratinocytes and fibroblasts in ways consistent with enhancement of the wound healing functions of all three cell types. We further show that 6, 8 dithio-UA significantly accelerates the wound healing process when applied topically (once daily) to full-thickness wounds in mice. Levels of Cu/Zn superoxide dismutase were increased in wound tissue from mice treated with 6, 8 dithio-UA compared to vehicle-treated mice, suggesting that the UA analog enhances endogenous cellular antioxidant defenses. These results support an adverse role for oxidative stress in wound healing and tissue repair, and provide a rationale for the development of UA analogs in the treatment of wounds and for modulation of angiogenesis in other pathological conditions.


Sensitivity of VCD spectroscopy for small structural and stereochemical changes of macrolide antibiotics.
We report the application of VCD spectroscopy for the characterization of clarithromycin and erythromycin. We show that the VCD spectra of these large macrolides are distinctly different and that spectra calculations reproduce the experimentally observed VCD signatures. In addition, computed VCD spectra of different epimers indicate that they should also be distinguishable from the correct structure of clarithromycin.


Outside management firm may help drive up parking revenues.
Larger healthcare organizations may be able to realize additional net revenues by using an outside firm to manage their paid-parking programs. As the author of this article explains, experienced outside management firms work to reduce operational expenditures and improve collections. They use program-specific revenue control methods and monitor parking activity on a daily, weekly, and monthly basis. Arrangements with outside parking management firms span a variety of lease and contract arrangements.


Free coracoacromial ligament graft for augmentation of massive rotator cuff tears treated with mini-open repair.
Repair of massive rotator cuff tears is a challenging and complex procedure. The tissue at the end of the torn tendons is often friable and weak, because ruptures are old and degenerated, and thus the ends must be sutured effectively and strongly to achieve a satisfactory outcome. We aimed to evaluate the effectiveness of free total or partial coracoacromial ligament (CAL) graft to protect and augment the repair of massive rotator cuff tears. Forty six patients ((32 females, mean age 54.3 years (range 39-66 years)) operated for massive rotator cuff tears between January 2003 and June 2009 were included in the study. Twenty nine of these patients had right-sided tears, and 17 had left-sided tears. Fifteen of the tears were 3-4 cm wide (mean 3.5 cm); 27 were >4 cm wide (mean 4.5 cm), and 4 were >5 cm wide. The mean follow-up period was 26 months (range 16-52 months). Patients were operated with a mini-open technique, and reconstructed after primary repair with 18 total and 28 partial free transfer of the CAL. Patients were evaluated by Constant-Murley score, and the degree of active flexion and abduction. Tendon thickness was measured with ultrasonography during follow-up. Mean preoperative shoulder flexion was 27.5° (range 5-40°), and mean abduction was 22.5° (range 10-30°). Shoulder flexion was significantly greater postoperatively (mean 102.6°, range 70-150°), as was shoulder abduction (mean 96.5°, range 60-150°). Mean preoperative and postoperative Constant-Murley score was 45 and 80, respectively. Surgical complications, particularly recurrence, did not occur in any patient during the follow-up period. The integrity and tendon thickness of the repairs were similar to those of normal tendons at the end of follow-up. Augmentation with a free transfer of the coracoacromial ligament provides excellent and promising functional results in the operative treatment of massive rotator cuff tears with a mini-open technique.


Co-Occurrence of Lower Urinary Tract Symptoms and Frailty among Community-Dwelling Older Men.
To estimate associations between lower urinary tract symptoms (LUTS) and phenotypic frailty in older men. Cross-sectional study. Community-dwelling men recruited from 2000 to 2002 from six U.S. academic centers for the Osteoporotic Fractures in Men Study. A total of 5,979 men aged 65 and older. The independent variable was LUTS severity (none/mild, moderate, or severe) assessed with the American Urologic Association Symptom Index. Participants were categorized as frail, intermediate stage, or robust using an adapted Cardiovascular Health Study index (components: low lean mass, weakness, exhaustion, slowness, and low physical activity). Associations were estimated with odds ratios and 95% confidence intervals (CIs) from multivariable multinomial logistic regression models adjusted for potential confounders of age, other demographics, health-related behaviors, and comorbidities. The prevalence of frailty was 7%, 11%, and 18% among men with none/mild, moderate, and severe LUTS, respectively. Moderate and severe LUTS, overall and by storage and voiding subscores, were associated with higher odds of both intermediate stage and frailty in all models. After adjustment for confounders, the odds of frailty was 1.41 times higher among men with moderate LUTS (95% CI = 1.14-1.74) and 2.51 times higher among men with severe LUTS (95% CI = 1.76-3.55), compared with none/mild LUTS. Severe LUTS was associated with a greater odds of individual frailty components exhaustion and low physical activity. The prevalence of phenotypic frailty is higher among older community-dwelling men with moderate or severe LUTS compared with those with mild or no LUTS. The positive association between LUTS severity and frailty among older men appears independent of age and known frailty risk factors. Although the temporal direction of this association and the utility of LUTS or frailty interventions in this population remain unclear, the high co-occurrence of these conditions could lead to earlier identification of frailty when clinically appropriate.


Biosynthetic incorporation of unnatural sialic acids into polysialic acid on neural cells.
In this study we demonstrate that polysialyltransferases are capable of accepting unnatural substrates in terminally differentiated human neurons. Polysialyltransferases catalyze the glycosylation of the neural cell adhesion molecule (NCAM) with polysialic acid (PSA). The unnatural sialic acid analog, N-levulinoyl sialic acid (SiaLev), was incorporated into cell surface glycoconjugates including PSA by the incubation of cultured neurons with the metabolic precursor N-levulinoylmannosamine (ManLev). The ketone group within the levulinoyl side chain of SiaLev was then used as a chemical handle for detection using a biotin probe. The incorporation of SiaLev residues into PSA was demonstrated by protection from sialidases that can cleave natural sialic acids but not those bearing unnatural N-acyl groups. The presence of SiaLev groups on the neuronal cell surface did not impede neurite outgrowth or significantly affect the distribution of PSA on neuronal compartments. Since PSA is important in neural plasticity and development, this mechanism for modulating PSA structure might be useful for functional studies.


Effect of bacteremia and fluid resuscitation on cardiac output.
The cardiac output acutely inducible by a 1-ml/kg/min saline infusion was observed in 12 septic and 7 nonseptic cynomolgus monkeys. In septic monkeys intravenous live E. coli were titrated to rapidly produce equivalent hypotension which was maintained for 1 hr before saline. Bacteria dose range was 1.3 X 10(11) bacteria/kg-400 X 10(11) bacteria/kg. There was no significant correlation between the dose of bacteria and the cardiac output nadir; however, the subsequent maximum cardiac output during saline resuscitation was significantly negatively correlated with the bacterial dose. In conclusion, widely varying amounts of intravenous bacteria can produce the same degree of hypotension and not correlate with cardiac output. In contrast, cardiac output measured early during subsequent saline resuscitation better predicts the severity of bacteremia.


Large, open datasets for human connectomics research: Considerations for reproducible and responsible data use.
Large, open datasets have emerged as important resources in the field of human connectomics. In this review, the evolution of data sharing involving magnetic resonance imaging is described. A summary of the challenges and progress in conducting reproducible data analyses is provided, including description of recent progress made in the development of community guidelines and recommendations, software and data management tools, and initiatives to enhance training and education. Finally, this review concludes with a discussion of ethical conduct relevant to analyses of large, open datasets and a researcher's responsibility to prevent further stigmatization of historically marginalized racial and ethnic groups. Moving forward, future work should include an enhanced emphasis on the social determinants of health, which may further contextualize findings among diverse population-based samples. Leveraging the progress to date and guided by interdisciplinary collaborations, the future of connectomics promises to be an impressive era of innovative research, yielding a more inclusive understanding of brain structure and function.


STAG1 mutations cause a novel cohesinopathy characterised by unspecific syndromic intellectual disability.
Cohesinopathies are rare neurodevelopmental disorders arising from a dysfunction in the cohesin pathway, which enables chromosome segregation and regulates gene transcription. So far, eight genes from this pathway have been reported in human disease. STAG1 belongs to the STAG subunit of the core cohesin complex, along with five other subunits. This work aimed to identify the phenotype ascribed to STAG1 mutations. Among patients referred for intellectual disability (ID) in genetics departments worldwide, array-comparative genomic hybridisation (CGH), gene panel, whole-exome sequencing or whole-genome sequencing were performed following the local diagnostic standards. A mutation in STAG1 was identified in 17 individuals from 16 families, 9 males and 8 females aged 2-33 years. Four individuals harboured a small microdeletion encompassing STAG1; three individuals from two families had an intragenic STAG1 deletion. Six deletions were identified by array-CGH, one by whole-exome sequencing. Whole-exome sequencing found de novo heterozygous missense or frameshift STAG1 variants in eight patients, a panel of genes involved in ID identified a missense and a frameshift variant in two individuals. The 17 patients shared common facial features, with wide mouth and deep-set eyes. Four individuals had mild microcephaly, seven had epilepsy. We report an international series of 17 individuals from 16 families presenting with syndromic unspecific ID that could be attributed to a STAG1 deletion or point mutation. This first series reporting the phenotype ascribed to mutation in STAG1 highlights the importance of data sharing in the field of rare disorders.


Specialization of plant-pollinator interactions increases with temperature at Mt. Kilimanjaro.
Species differ in their degree of specialization when interacting with other species, with significant consequences for the function and robustness of ecosystems. In order to better estimate such consequences, we need to improve our understanding of the spatial patterns and drivers of specialization in interaction networks. Here, we used the extensive environmental gradient of Mt. Kilimanjaro (Tanzania, East Africa) to study patterns and drivers of specialization, and robustness of plant-pollinator interactions against simulated species extinction with standardized sampling methods. We studied specialization, network robustness and other network indices of 67 quantitative plant-pollinator networks consisting of 268 observational hours and 4,380 plant-pollinator interactions along a 3.4 km elevational gradient. Using path analysis, we tested whether resource availability, pollinator richness, visitation rates, temperature, and/or area explain average specialization in pollinator communities. We further linked pollinator specialization to different pollinator taxa, and species traits, that is, proboscis length, body size, and species elevational ranges. We found that specialization decreased with increasing elevation at different levels of biological organization. Among all variables, mean annual temperature was the best predictor of average specialization in pollinator communities. Specialization differed between pollinator taxa, but was not related to pollinator traits. Network robustness against simulated species extinctions of both plants and pollinators was lowest in the most specialized interaction networks, that is, in the lowlands. Our study uncovers patterns in plant-pollinator specialization along elevational gradients. Mean annual temperature was closely linked to pollinator specialization. Energetic constraints, caused by short activity timeframes in cold highlands, may force ectothermic species to broaden their dietary spectrum. Alternatively or in addition, accelerated evolutionary rates might facilitate the establishment of specialization under warm climates. Despite the mechanisms behind the patterns have yet to be fully resolved, our data suggest that temperature shifts in the course of climate change may destabilize pollination networks by affecting network architecture.


Intestinal parasitism, potable water availability and methods of sewage disposal in three communities in Benue State, Nigeria: a survey.
To assess the level of parasite burden in a village community and the predisposing factors. Two hundred subjects each were recruited from three communities- Tyogbenda, Jato-Aka and Adikpo during an episode of free medical outreach. A simple random sampling method was adopted and a questionnaire was interviewer administered on relevant aspects of basic hygiene such as- sources of water supply, methods of domestic sewage disposal and frequency of hand washing. Stool samples were collected and tested and findings analysed using appropriate statistical methods, p values < 0.05 were considered significant. The prevalence of intestinal parasites in Tyogbenda, Jato-Aka and Adikpo communities was found to be 71.5%, 65.5% and 40.5% respectively. Ascaris lumbricoides was the commonest parasite in the three centres (34.5%, 28.5% and 19.0% respectively for Tyogbenda, Jato-Aka and Adikpo communities). Other parasites identified were- Entamoeba histolytica, Entamoeba coli, Hookworm. Enterobius vermicularis, Strongyloides stercoralis, Schistosoma mansoni and Trichuris trichura. Multiple parasitic infestation was common in the communities where quality of water supply and methods of sewage disposal facilities were below standard. The prevalence of intestinal parasitism is still high in Nigerian rural communities. The present resolve by the federal ministry of water resources to supply potable water to all Nigerian rural communities should be sustained. Furthermore, a health education program should be properly constituted and integrated into the present primary health care policy for the country.


Exercise-Related Physical Activity Relates to Brain Volumes in 10,125 Individuals.
The potential neuroprotective effects of regular physical activity on brain structure are unclear, despite links between activity and reduced dementia risk. To investigate the relationships between regular moderate to vigorous physical activity and quantified brain volumes on magnetic resonance neuroimaging. A total of 10,125 healthy participants underwent whole-body MRI scans, with brain sequences including isotropic MP-RAGE. Three deep learning models analyzed axial, sagittal, and coronal views from the scans. Moderate to vigorous physical activity, defined by activities increasing respiration and pulse rate for at least 10 continuous minutes, was modeled with brain volumes via partial correlations. Analyses adjusted for age, sex, and total intracranial volume, and a 5% Benjamini-Hochberg False Discovery Rate addressed multiple comparisons. Participant average age was 52.98±13.04 years (range 18-97) and 52.3% were biologically male. Of these, 7,606 (75.1%) reported engaging in moderate or vigorous physical activity approximately 4.05±3.43 days per week. Those with vigorous activity were slightly younger (p < 0.00001), and fewer women compared to men engaged in such activities (p = 3.76e-15). Adjusting for age, sex, body mass index, and multiple comparisons, increased days of moderate to vigorous activity correlated with larger normalized brain volumes in multiple regions including: total gray matter (Partial R = 0.05, p = 1.22e-7), white matter (Partial R = 0.06, p = 9.34e-11), hippocampus (Partial R = 0.05, p = 5.96e-7), and frontal, parietal, and occipital lobes (Partial R = 0.04, p≤1.06e-5). Exercise-related physical activity is associated with increased brain volumes, indicating potential neuroprotective effects.


A genomics approach reveals the global genetic polymorphism, structure, and functional diversity of ten accessions of the marine model diatom Phaeodactylum tricornutum.
Diatoms emerged in the Mesozoic period and presently constitute one of the main primary producers in the world's ocean and are of a major economic importance. In the current study, using whole genome sequencing of ten accessions of the model diatom Phaeodactylum tricornutum, sampled at broad geospatial and temporal scales, we draw a comprehensive landscape of the genomic diversity within the species. We describe strong genetic subdivisions of the accessions into four genetic clades (A-D) with constituent populations of each clade possessing a conserved genetic and functional makeup, likely a consequence of the limited dispersal of P. tricornutum in the open ocean. We further suggest dominance of asexual reproduction across all the populations, as implied by high linkage disequilibrium. Finally, we show limited yet compelling signatures of genetic and functional convergence inducing changes in the selection pressure on many genes and metabolic pathways. We propose these findings to have significant implications for understanding the genetic structure of diatom populations in nature and provide a framework to assess the genomic underpinnings of their ecological success and impact on aquatic ecosystems where they play a major role. Our work provides valuable resources for functional genomics and for exploiting the biotechnological potential of this model diatom species.


Aberrant TCR-mediated signaling in CD45-null thymocytes involves dysfunctional regulation of Lck, Fyn, TCR-zeta, and ZAP-70.
CD45 is a transmembrane phosphotyrosine phosphatase expressed on all nucleated hemopoietic cells. Targeting of CD45 exon 9 has generated a mouse line completely lacking CD45 expression (CD45-null) in which there are severe abnormalities in T cell development. Defects in TCR-mediated signals underlying these abnormalities have now been investigated using CD45-null T cells. No T cell proliferation was detected in response to a CD3 mAb. In thymocytes the p56(lck) and p59(fyn) tyrosine kinases were hyperphosphorylated, and p56(lck) was in its inactive conformation. Both basal and TCR-stimulated tyrosine phosphorylation of TCR-zeta and CD3-epsilon were much reduced, and TCR stimulation induced an abnormal p18 phosphoisomer of TCR-zeta previously noted in T cells stimulated by altered peptide ligands. These defects were associated with the failure of ZAP-70 kinase recruitment to the TCR-zeta chain. TCR coupling to the tyrosine phosphorylation of several proteins, including HS1 and p120(cbl), was also much reduced. However, TCR-induced signaling was not ablated, and significant inositol phosphate and calcium signals were observed in CD45-null thymocytes. Our molecular analysis suggests that the threshold for TCR signal transduction is greatly increased in CD45-null T cells, thus explaining the profound defects in thymic development.


Outcomes of Right Ventricular Outflow Tract Reconstruction in Children: Retrospective Comparison Between Bovine Jugular Vein and Expanded Polytetrafluoroethylene Conduits.
Bovine jugular vein (BJV) and expanded polytetrafluoroethylene (ePTFE) conduits have been described as alternatives to the homograft for right ventricular outflow tract (RVOT) reconstruction. This study compared RVOT reconstructions using BJV and ePTFE conduits performed in a single institution. The valve functions and outcomes of patients aged < 18 years who underwent primary RVOT reconstruction with a BJV or ePTFE conduit between 2013 and 2017 were retrospectively investigated. 44 patients (20 and 24 with BJV and ePTFE conduits, respectively) met the inclusion criteria. The mean follow-up time was 4.5 ± 1.5 years. No significant differences in peak RVOT velocity (1.8 ± 0.9 m/s vs 2.1 ± 0.9 m/s, P = 0.27), branch pulmonary stenosis (P = 0.50), or pulmonary regurgitation (P = 0.44) were found between the BJV and ePTFE conduit groups, respectively. Aneurysmal dilatation of the conduit was observed in 25.0% of the patients in the BJV conduit group but not in the ePTFE conduit group (P = 0.011). All the cases with aneurysmal dilatation of the BJV conduit were complicated with branch pulmonary stenosis up to 3.0 m/s (P = 0.004). No conduit infections occurred during the follow-up period, and no significant difference in conduit replacement (20.0% vs 8.3%, P = 0.43) was found between the BJV and ePTFE conduit groups, respectively. The outcomes of the RVOT reconstructions with BJV and ePTFE conduits were clinically satisfactory. Aneurysmal dilatation was found in the BJV conduit cases, with branch pulmonary stenosis as the risk factor.


Telomerase activity in benign and malignant thyroid diseases.
Telomerase, an enzyme associated with cellular immortality, is expressed by most malignant cells and is inactive in most normal somatic cells, with the excitation of proliferative stem cells, male germ cells, and activated lymphocytes. The measurement of telomerase activity in clinically obtained tissue samples may provide useful information as both a diagnostic and prognostic marker. In this study, we sought to determine whether telomerase activity might prove helpful in the assessment of benign and malignant thyroid tumors. A modified, semiquantitative polymerase chain reaction-based telomeric repeat amplification protocol assay was used for detection of telomerase activity in 59 samples obtained at thyroidectomy, including 15 thyroid cancers, 22 benign thyroid diseases, and 22 adjacent normal thyroid tissues. Four of 13 differentiated thyroid carcinomas (30%) and 2 of 2 medullary carcinomas (100%) expressed telomerase activity. Unexpectedly, we also detected activity in 3 of 22 (14%) adjacent normal thyroid tissues and 6 of 22 (28%) benign thyroid diseases. Pathologic review of the telomerase-positive benign specimens revealed that many contained extensive lymphoid infiltrates with germinal centers (six of nine, 67%), as did two of four telomerase-positive papillary carcinomas. In contradistinction to other epithelial carcinomas, telomerase does not appear to be frequently reactivated in differentiated thyroid carcinomas.


Health economics methods for public health resource allocation: a qualitative interview study of decision makers from an English local authority.
Local authorities in England have responsibility for public health, however, in recent years, budgets have been drastically reduced placing decision makers under unprecedented financial pressure. Although health economics can offer support for decision making, there is limited evidence of it being used in practice. The aim of this study was to undertake in-depth qualitative research within one local authority to better understand the context for public health decision making; what, and how economics evidence is being used; and invite suggestions for how methods could be improved to better support local public health decision making. The study included both observational methods and in-depth interviews. Key meetings were observed and semi-structured interviews conducted with participants who had a decision-making role to explore views on economics, to understand the barriers to using evidence and to invite suggestions for improvements to methods. Despite all informants valuing the use of health economics, many barriers were cited: including a perception of a narrow focus on the health sector; lack of consideration of population impact; and problems with translating long timescales to short term impact. Methodological suggestions included the broadening of frameworks; increased use of natural experiments; and capturing wider non-health outcomes that resonate with the priorities of multiple stakeholders.


Investigation of the C242T polymorphism of NAD(P)H oxidase p22 phox gene and ischaemic heart disease using family-based association methods.
Ischaemic heart disease is a complex phenotype arising from the interaction of genetic and environmental factors. Excessive production of reactive oxygen species leading to endothelial dysfunction is believed to be important in the pathogenesis of ischaemic heart disease. The NAD(P)H oxidase system generates superoxide anions in vascular cells; however, the role of the C242T polymorphism of the NAD(P)H oxidase p22 phox gene in ischaemic heart disease is unclear due to contradictory results from case-control studies. Consequently, we applied family-based association tests to investigate the role of this polymorphism in ischaemic heart disease in a well-defined Irish population. A total of 1023 individuals from 388 families (discordant sibships and parent/child trios) were recruited. Linkage disequilibrium between the polymorphism and ischaemic heart disease was tested using the combined transmission disequilibrium test (TDT)/sib-TDT (cTDT) and pedigree disequilibrium test (PDT). Both cTDT and PDT analyses found no statistically significant excess transmission of either allele to affected individuals (P =0.30 and P =0.28, respectively). Using robust family-based association tests specifically designed for the study of complex diseases, we found no evidence that the C242T polymorphism of the p22 phox gene has a significant role in the development of ischaemic heart disease in our population.


[Armauer Hansen (1841-1912), portrait of a Nordic pioneer].
Desending from a Danish family, Gerhard Henrik Armauer Hansen was born in Bergan, Norway, on 29 July, 1841. Graduated in medicine in 1866 from the University at Christiana (the former name of Oslo), he started his medical career as an assistant physician at the National Hospital of Oslo for one year, working afterwards as a medical officer for a fishing company outside the Arctic Circle. Taking not well his routine job, he soon returned to Bergen where he began his work on a disease known as leprosy at the age of 26 and as an assistant of D.C. Danielssen at the Lungegaarden Hospital. There he engaged in research and investigations on the nature of the disease. Studying the various types of leprosy (skin, nerve and visceral lesions), he was awarded, one year after beginning his work, the gold metal of the University for his first publication on the disease. While Danielssen leaned toward heredity as a dominant factor in leprosy, Hansen's conviction was that the disease must have an infectious causal agent. In about 1871, Hansen began to observe tiny little rods in tissue specimens and considered they could be the ethiologic agent of leprosy, the more he found these rods in all the infiltrated nodular lesions in his patients. Finally, he proposed on February 28, 1873, that the rods were bacilli, responsible of leprosy. Promoted to the rank of Chief of the Leprosy Service in 1875, he held his job for 37 years; Hansen's entire adult life was spent at work in leprosy. He edited the journal <<Lepra>>. Hansen was also an eminent zoologist engaged in studies involving mollusks and worms; Since 1874, he was president of the Bergen Museum of Natural History. Armauer Hansen died on February 12, 1912, and the funeral ceremonies took place in the Museum of Bergen where his ashes are still kept.


[The accumulation of different substituted hydroxyethyl starches (HES) following repeated infusions in healthy volunteers].
Accumulation of hydroxyethyl starch (HES) after repeated applications of starches with different molar substitution and similar molecular weight was investigated. Treatment with five consecutive infusions of hydroxyethyl starch was carried out using two medium molecular weight hydroxyethyl starches with a molar substitution of 0.5 and 0.62. Healthy volunteers received 500 ml 6% HES 200/0.62 (30 g) or 500 ml 10% HES 200/0.5 per day over a period of five consecutive days. Blood samples were taken in the morning before infusions were started (7.A.M.) and at each hour during the infusion period of 4 hours post infusionem until 4 hours after the infusions. During the first 10 days and on the 20th and on the 30th day after the last infusion blood samples were taken. Both HES solutions were subjectively and objectively well tolerated by healthy volunteers. No side effects were observed. However, pharmacokinetics of the investigated HES-formulations were significantly different. The model-independent calculated elimination half life time (T1/2) increased from day to day. During the five days T1/2 was prolonged for 20 h by high substituted HES (200/0.62) and for 2.5 h for medium substituted HES (200/0.5). The half life times related to the three compartment model calculation were with 0.6 h, 11.6 h and 211 h for HES 200/0.62 two fold higher than the times for HES 200/0.5 with 0.39 h, 6.98 h and 113 h. Plasma clearance for HES 200/0.5 (4.86 ml/min) was five fold higher than that from HES 200/0.62 with 0.98 ml/min. With the exception of the first day of infusion serum concentrations of HES 200/0.5 although only 30 g HES 200/0.6 versus 50 g HES 200/0.5 were infused. No difference of the hemodilution effects between the two HES-formulations were observed. The hemorheologic parameters were similar in both groups with the exception of plasma viscosity which was significantly higher after infusion of HES 200/0.62. High substituted HES accumulate in serum more than medium substituted HES. Especially when HES must be applied in multiple doses, high substituted HES should not be used or the infusion interval must be adapted to the elimination half life time of the used HES.


Genetics of flavonoid, carotenoid, and chlorophyll pigments in melon fruit rinds.
External color has profound effects on acceptability of agricultural products by consumers. Carotenoids and chlorophylls are known to be the major pigments of melon (Cucumis melo L.) rinds. Flavonoids (especially chalcones and anthocyanins) are also prominent in other fruits but have not been reported to occur in melons fruit. We analyzed the pigments accumulating in rinds of different melon genotypes during fruit development. We found that melon rind color is based on different combinations of chlorophyll, carotenoids, and flavonoids according to the cultivar tested and their ratios changed during fruit maturation. Moreover, in "canary yellow" type melons, naringenin chalcone, a yellow flavonoid pigment previously unknown to occur in melons, has been identified as the major fruit colorant in mature rinds. Naringenin chalcone is also prominent in other melon types, occurring together with carotenoids (mainly β-carotene) and chlorophyll. Both chlorophyll and carotenoid pigments segregate jointly in an F(2) population originating from a cross between a yellow canary line and a line with green rind. In contrast, the content of naringenin chalcone segregates as a monogenic trait independently to carotenoids and chlorophyll. Transcription patterns of key structural phenylpropanoid and flavonoid biosynthetic pathway genes were monitored in attempts to explain naringenin chalcone accumulation in melon rinds. The transcript levels of CHI were low in both parental lines, but C4H, C4L, and CHS transcripts were upregulated in "Noy Amid", the parental line that accumulates naringenin chalcone. Our results indicate that naringenin chalcone accumulates independently from carotenoids and chlorophyll pigments in melon rinds and gives an insight into the molecular mechanism for the accumulation of naringenin chalcone in melon rinds.


Critical comparison of the on-line and off-line molecularly imprinted solid-phase extraction of patulin coupled with liquid chromatography.
Reaching trace amounts of mycotoxin contamination requires sensitive and selective analytical tools for their determination. Improving the selectivity of sample pretreatment steps covering new and modern extraction techniques is one way to achieve it. Molecularly imprinted polymers as selective sorbent for extraction undoubtedly meet these criteria. The presented work is focused on the hyphenation of on-line molecularly imprinted solid-phase extraction with a chromatography system using a column-switching approach. Making a critical comparison with a simultaneously developed off-line extraction procedure, evaluation of pros and cons of each method, and determining the reliability of both methods on a real sample analysis were carried out. Both high-performance liquid chromatography methods, using off-line extraction on molecularly imprinted polymer and an on-line column-switching approach, were validated, and the validation results were compared against each other. Although automation leads to significant time savings, fewer human errors, and required no handling of toxic solvents, it reached worse detection limits (15 versus 6 μg/L), worse recovery values (68.3-123.5 versus 81.2-109.9%), and worse efficiency throughout the entire clean-up process in comparison with the off-line extraction method. The difficulties encountered, the compromises made during the optimization of on-line coupling and their critical evaluation are presented in detail.


Electroconvulsive shock: progressive dopamine autoreceptor subsensitivity independent of repeated treatment.
Repeated electroconvulsive shock, applied to rats, induces a subsensitivity of dopamine autoreceptors located in the substantia nigra as indexed by single-unit electrophysiological techniques. This reduced sensitivity is time-dependent, since effects similar to those seen with repeated treatment were also observed when single electroconvulsive shock was followed by an appropriate treatment-free interval. These data, coupled with identical results after the repeated administration of tricyclic antidepressants, raise the possibility that a reduction of dopamine autoreceptor sensitivity could underlie both electroconvulsive shock and pharmacological treatment of depression.


Exploring preferences of market traders on the type and delivery methods of HIV services in Lilongwe, Malawi.
Background: Reaching all people with HIV services, including traders in the informal economy, is critical to meeting UNAIDS' 95-95-95 goals. However, traders prioritise their business over attendance at health facilities. This limits their access to health services. This study explores market traders' preferences for the potential type and delivery methods of HIV services at Lilongwe Central market.Method: The study used an exploratory qualitative study design in Lilongwe, Malawi. Sixteen in-depth interviews were conducted among traders at Lilongwe Central Market between June and September 2022. In the same period, we also conducted four key informant interviews involving three officers responsible for HIV services at the district and council levels, and the market chairman.Results: HIV services preferred by market traders include HIV testing, antiretroviral therapy, condom dispensation, voluntary medical male circumcision and HIV awareness campaigns. These services should be offered daily or when the market is less crowded, and they could be delivered in the market. These services can be provided by both lay and health workers, depending on traders' preferences, and must be integrated with other health services to mitigate unintended HIV status disclosure concerns.Conclusion: The achievement of UNAIDS' 95-95-95 goals by 2030 requires that HIV services should be available to all those who require them at times and locations that are convenient for them, through providers they have chosen either as integrated or standalone, depending on the target group perception of the role of these two models in mitigating stigma. This will necessitate the development of new approaches targeting underserved groups, such as traders in markets.


[Circahoral fluctuations in protein release from hepatocytes in a monolayer culture].
An account is presented on morphological and functional properties in primary monolayer culture. Advantages of this model, as compared to the suspension cells, for studying various aspects of the liver vital activity are described. Dynamics of protein secretion by hepatocytes during a 2-hour period was investigated by different methods. The data obtained point to cyclic changes of the rate of protein secretion in the liver cells with circahoralian periodicity, which is typical of the changes in the rate of protein synthesis we revealed earlier for the same cells.


Novel gas producing Vibrio cholerae: a case report of gastroenteritis with acute kidney injury.
Bacterial characterization is important in clinical and epidemiological studies. We herein report the first case of gas-producing Vibrio cholera gastroenteritis with acute kidney injury. A 30-year-old female presented to the emergency department with complaints of about ten episodes of watery diarrhea, four episodes of vomiting and elevated serum urea/creatinine levels. Although the bacteria were first misidentified as Vibrio furnissii by gas production on carbohydrate fermentation and triple sugar iron agar, it was later confirmed as Vibrio cholerae by 16 S rRNA gene sequencing and specific PCR. The treatment regimen was followed as for Vibrio species with intravenous fluids, ciprofloxacin and doxycycline. The patient recovered without relapse. Literature survey from the PubMed database shows no gas-producing Vibrio cholerae isolate being reported in the world. Further, genotype studies are warranted to look into the gas production of Vibrio cholerae .


An Optimum Fatigue Design of Polymer Composite Compressed Natural Gas Tank Using Hybrid Finite Element-Response Surface Methods.
The main purpose of this research is to design a high-fatigue performance hoop wrapped compressed natural gas (CNG) composite cylinder. To this end, an optimization algorithm was presented as a combination of finite element simulation (FES) and response surface analysis (RSA). The geometrical model was prepared as a variable wall-thickness following the experimental measurements. Next, transient dynamic analysis was performed subjected to the refueling process, including the minimum and maximum internal pressures of 20 and 200 bar, respectively. The time histories of stress tensor components were extracted in the critical region. Furthermore, RSA was utilized to investigate the interaction effects of various polymer composite shell manufacturing process parameters (thickness and fiber angle) on the fatigue life of polymer composite CNG pressure tank (type-4). In the optimization procedure, four parameters including wall-thickness of the composite shell in three different sections of the CNG tank and fiber angle were considered as input variables. In addition, the maximum principal stress of the component was considered as the objective function. Eventually, the fatigue life of the polymer composite tank was calculated using stress-based failure criterion. The results indicated that the proposed new design (applying optimal parameters) leads to improve the fatigue life of the polymer composite tank with polyethylene liner about 2.4 times in comparison with the initial design.


Modulating mesenchymal stromal cell function with cholesterol synthesis inhibitors.
There is increasing evidence that statins, inhibitors of 3-hydroxyl-3-methylglutaryl coenzyme A reductase, can effectively be used not only in the treatment of hypercholesterolemia, but also in other human disorders; indeed, statins have strong anti-inflammatory and immunomodulatory effects, so that they can influence the onset and outcome of inflammation and autoimmunity. On the other hand, it has been shown that statins can affect growth and survival of solid tumour and leukemic cells, thus they have been proposed in the treatment of neoplasias as multiple myeloma, in association with drugs, as thalidomide, known to act on the cancer microenvironment. In the current view, tumor microenvironment include many cell types that interact with tumor cells: among them, stromal and endothelial cells, macrophages and dendritic cells, the various types of lymphocytes such as NK cells, B and T cells. The interplay between all these cell populations, and the balance between these, determines whether there is a tumour cell growth promotion or inhibition. In haematological malignancies, such as multiple myeloma, chronic lymphocytic and myeloid leukemias and follicular lymphomas, the survival, drug-resistance and proliferation of leukemic cells have been shown to be largely dependent on a supportive microenvironment, so that some cellular components of it, mainly mesenchymal stromal cells, cancer associated fibroblasts and macrophages, are now proposed as targets of new therapies. Herein, we analyze the effects that statins can exert on cancer cells, stromal cells and human natural killer cells, to discuss whether they can be proposed as anti-cancer drugs.


Host response to coagulase-negative staphylococci in abscesses induced within mice.
A model whereby a known number of coagulase-negative staphylococci were packed into capillary tubes and implanted into the peritoneal cavity of mice proved to be a satisfactory method for generating abscesses that could be easily removed free of extraneous host tissue, and that permitted measurement of the survival of the organisms and accumulation of lipid in the lesion. Strains of S. epidermidis, S. schleiferi and S. lugdunensis, differing in their ability to produce fatty acid modifying enzyme (FAME) and lipase, were packed into either glass or plastic capillary tubes and used to generate abscesses. Abscesses produced by S. aureus served as comparators. Lipids accumulated within the abscesses caused by S. aureus in the same manner as previously described for the organism inoculated without tubes. Lipids also accumulated within abscesses produced by all the coagulase-negative staphylococci, but the rate of accumulation was slower and the lipid droplets were smaller than seen with S. aureus. The mobilisation of lipid did not differ in response to cocci in plastic or glass tubes. Strains of S. epidermidis and S. schleiferi producing FAME and lipase were better able to survive within abscesses than strains unable to produce these enzymes. However, FAME and lipase production did not appear to be the sole determinants of survival within abscesses. Regardless of whether they produced FAME and lipase, the two S. epidermidis strains were significantly better able to survive within plastic tubes than in glass tubes. No such difference was seen with S. aureus between plastic and glass tubes.


Wind chill reconsidered, Siple revisited.
Experiments similar to those establishing the concepts of wind chill have been done with bare and "clothed" cylinders and when the clothing was wet at temperatures above and below freezing. Clothing prevented the heat loss that was associated with increased wind velocity in the bare cylinder. With wet clothing, evaporation increased with wind velocity but heat loss did not, and raincoats or freezing markedly reduced wet heat loss. The concept of wind chill applies only to unprotected objects.


Observation of disorder-induced 2D mott-hubbard states of the alkali-earth metal (Mg,Ba)-adsorbed Si(111) surface
We report evidence of a disorder-driven Mott-Hubbard-type localization on the alkali-earth metal (AEM) (Mg,Ba)-adsorbed Si(111)-(7x7) surface. The clean metallic Si(111) surface is found to undergo a two-dimensional (2D) metal-insulator transition as randomly distributed AEM adsorbates cause disorder on the surface. A well-defined electron-energy-loss peak unique to the insulating phase is attributed to an interband excitation between the split Hubbard bands originated from a metallic surface band at Fermi energy. A quantitative analysis of the loss peak reveals that the AEM-induced insulating surfaces are of a Mott-Hubbard type driven essentially by disorder.


Fabrication of Integrated Cu2O@HKUST-1@Au Nanocatalysts via Galvanic Replacements toward Alcohols Oxidation Application.
Metastable Cu2O is an attractive material for the architectural design of integrated nanomaterials. In this context, the in situ growth of microporous metal-organic frameworks (MOFs) on Cu2O nanocrystals with well-defined shapes (such as octahedron, truncated octahedron, and corner-truncated cube) was conducted under ambient conditions by using sacrificial Cu2O nanocrystals as a copper ion source for the construction of HKUST-1. It was found that the growth rate of HKUST-1 on the (111) facet of octahedral Cu2O was much faster than that on the (100) facet of cubic Cu2O. A subsequent etching process on the core-shell-structured Cu2O@HKUST-1 to remove Cu2O results in hollow HKUST-1 with hierarchical pores, where the shapes of cavities are tailored depending on the original Cu2O crystals. Importantly, it was found that the integrated Cu2O@HKUST-1@Au nanocatalysts could be fabricated by galvanic replacement reaction between Cu2O and AuCl4-, wherein the copper(I) ions from Cu2O etching diffuse through the microporous MOF shell and reach to the external surface to reduce AuCl4- ions, thereby forming Au nanoparticles exclusively on the MOF external surface. Furthermore, the catalytic applications of Cu2O@HKUST-1@Au toward liquid-phase oxidation of both aromatic and aliphatic alcohols were studied. Notably, 100% selectivities of aldehyde products were achieved without any further oxidation byproducts such as acid and ester.


Postprandial kinetics of digestive function in rainbow trout (Oncorhynchus mykiss): genes expression, enzymatic activity and blood biochemistry as a practical tool for nutritional studies.
Postprandial kinetics of genes expression of gastric (chitinase, pepsinogen) and intestinal (alkaline phosphatase, maltase) digestive enzymes and nutrient transporters (peptide transporter 1, sodium-glucose transporter 1), Brush Border Membrane (BBM) enzymes activity (alkaline phosphatase, leucine aminopeptidase, maltase, saccharase) and blood biochemistry (triglycerides, cholesterol, protein, albumin, glucose, amino acids) through NMR spectroscopy, were investigated in rainbow trout (Oncorhynchus mykiss) fed a commercial aquafeed. For this purpose, fish were starved 72 h and digestive tract and blood were sampled before the meal and at 1.5, 3, 6, 9, 12, and 24 h after feeding (T0, T1.5, T3, T6, T9, T12 and T24). The postprandial kinetic showed that the expression of the genes involved in digestion and nutrient transport, the activity of BBM enzymes, and the presence of metabolites in blood were stimulated in different ways by the presence of feed in the digestive tract. The expression of most genes peaked 3 h after meal except gastric pepsinogen and maltase in distal intestine that peaked at T9 and T12, respectively. The activity of BBM enzymes were stimulated differently based on the intestine tract. The plasma proteins level increased from T1.5 until T9, while the other blood parameters unvariated during the postprandial period. This study supplied useful information about the physiological effects a single meal as a potential tool for planning nutritional studies involving the digestive functions.


A population level analysis of second hematological malignancies in chronic lymphocytic leukemia/small lymphocytic lymphoma survivors in the era of targeted therapies.
With improvement in survival after chronic lymphocytic leukemia (CLL) diagnosis, the real-world burden of second hematological malignancies (SHM) has not been comprehensively assessed in recent era. We analyzed risk, incidence, and outcomes of SHM in CLL patients between 2000 and 2019 using SEER database. CLL patients had greater risk for hematological malignancies than general population [SIR, standardized incidence ratio (95% CI):2.58 (2.46-2.70); p < 0.05]. The risk for subsequent lymphoma increased by 1.75 folds in 2015-2019 compared to 2000-2004. The duration, after CLL diagnosis, of maximum risk for SHM decreased as 60-119 months for time-period 2000-2004, 6-11 months for 2005-2009 to 2-5 months for 2010-2014 and 2015-2019. Incidence of SHM was 2.5% in CLL survivors (1736/70,346) with lymphoid SHM being more common than myeloid SHM, and DLBCL being the most common pathology (n = 610, 35% of all SHM). Male sex, age ≤65 years at CLL diagnosis, and chemotherapy treatment were associated with higher risk for SHM. The median gap between CLL and SHM diagnoses was 46 months. The median survival for de-novo-AML, t-MN, CML, and aggressive NHL was 63, 86, 95, and 96 months respectively. Although SHM remains rare, there is increased risk in recent era, likely due to improved survival in CLL patients, necessitating active surveillance strategies.


First notification of positive blood cultures and the high accuracy of the gram stain report.
When blood cultures turn positive, the attending physicians are usually notified immediately about Gram stain findings. However, information on the accuracy of Gram staining is very limited. We examined the accuracy of preliminary blood culture reports provided by a regional laboratory in an observational study including the years 1996, 2000 to 2001, and 2003. We used data from computer files and technicians' laboratory notes. The study was restricted to cultures with one morphological type. Using cultural identification as a reference, we estimated the sensitivity, specificity, and positive and negative predictive values (PPV and NPV, respectively) for the following defined morphological groups: gram-positive cocci in clusters, gram-positive cocci in chains or diplococci, gram-positive rods, gram-negative cocci, gram-negative rods, and yeasts. We further evaluated the Gram stain and wet mount findings for the most frequent bacterial species/groups. We obtained 5,893 positive blood cultures and the following results for the defined groups: sensitivity, range of 91.3 to 99.7%; specificity, 98.9 to 100%; PPV, 94.6 to 100%; and NPV, 99.0 to 100%. The sensitivity for the most frequent species was in the range 91.3 to 100%, with nonhemolytic streptococci having the lowest value (sensitivity, 91.3%; 95% confidence interval, 86.2 to 94.9%). Wet mount reports were less accurate (sensitivity of 30 to 70% for species with peritrichous motility), and Enterobacteriaceae (notably Salmonella spp.) accounted for 25% of the reports stating polar motility. In conclusion, we demonstrated a high accuracy of Gram stain reports, whereas wet mount microscopy was generally less accurate.


Subjective Memory in a National Sample: Predicting Psychological Well-Being.
Individual perception of memory performance (i.e., subjective memory) is assessed using a variety of approaches. This article focuses on 2 such approaches: (1) self-comparison assessments that attempt to capture changes in memory ability over a period of time and (2) age-anchored comparisons that assess how an individual perceives their memory in relation to others their age. These different types of assessment may relate to psychological well-being differently due to the underlying mechanisms of assessment. The purpose of these analyses is to examine 2 measures of subjective memory (i.e., a self-comparison measure and an age-anchored comparison measure) as predictors of psychological well-being among adults in mid- and late life. Participants (n = 3,434) in the Midlife in the United States Study completed measures of subjective memory, depressive affect, and life satisfaction. Structural equation modeling was used to examine whether the self-comparison and age-anchored comparison measures had differential predictive utility regarding psychological well-being. Higher age-anchored comparison ratings were related to higher life satisfaction scores. There was a significant interaction between the 2 items such that individuals with lower ratings on both subjective memory measures had the poorest outcomes. Additionally, age-anchored comparisons interacted with age: older adults had the poorest outcomes when they reported poorer age-anchored comparisons. These findings highlight the importance of precise measurement in the consideration of subjective memory. How an individual was asked to rate his or her perception of memory influenced the relationships between subjective memory and psychological well-being. This study contributes valuable insight into the importance of the assessment models of subjective memory.


p54nrb/NonO and PSF promote U snRNA nuclear export by accelerating its export complex assembly.
The assembly of spliceosomal U snRNPs in metazoans requires nuclear export of U snRNA precursors. Four factors, nuclear cap-binding complex (CBC), phosphorylated adaptor for RNA export (PHAX), the export receptor CRM1 and RanGTP, gather at the m(7)G-cap-proximal region and form the U snRNA export complex. Here we show that the multifunctional RNA-binding proteins p54nrb/NonO and PSF are U snRNA export stimulatory factors. These proteins, likely as a heterodimer, accelerate the recruitment of PHAX, and subsequently CRM1 and Ran onto the RNA substrates in vitro, which mediates efficient U snRNA export in vivo. Our results reveal a new layer of regulation for U snRNA export and, hence, spliceosomal U snRNP biogenesis.


Intestinal alkaline phosphatase in the colonic mucosa of children with inflammatory bowel disease.
To investigate intestinal alkaline phosphatase (iAP) in the intestinal mucosa of children with inflammatory bowel disease (IBD). Colonic biopsy samples were taken from 15 newly diagnosed IBD patients and from 10 healthy controls. In IBD patients, specimens were obtained both from inflamed and non-inflamed areas. The iAP mRNA and protein expression was determined by reverse transcription-polymerase chain reaction and Western blotting analysis, respectively. Tissue localization of iAP and Toll-like receptor (TLR) 4 was investigated by immunofluorescent staining. The iAP protein level in the inflamed mucosa of children with Crohn's disease (CD) and ulcerative colitis (UC) was significantly decreased when compared with controls (both P < 0.05). Similarly, we found a significantly decreased level of iAP protein in the inflamed mucosa in CD compared with non-inflamed mucosa in CD (P < 0.05). In addition, the iAP protein level in inflamed colonic mucosa in patients with UC was decreased compared with non-inflamed mucosa in patients with CD (P < 0.05). iAP protein levels in the non-inflamed mucosa of patients with CD were similar to controls. iAP mRNA expression in inflamed colonic mucosa of children with CD and UC was not significantly different from that in non-inflamed colonic mucosa with CD. Expression of iAP mRNA in patients with non-inflamed mucosa and in controls were similar. Co-localization of iAP with TLR4 showed intense staining with a dotted-like pattern. iAP was present in the inflamed and non-inflamed mucosa of patients with CD, UC, and in control biopsy specimens, irrespective of whether it was present in the terminal ileum or in the colon. However, the fluorescent signal of TLR4 was more pronounced in the colon compared with the terminal ileum in all groups studied. Lower than normal iAP protein levels in inflamed mucosa of IBD patients may indicate a role for iAP in inflammatory lesions in IBD. Based on our results, administration of exogenous iAP enzyme to patients with the active form of IBD may be a therapeutic option.


Phytoremediation of aged petroleum sludge: effect of inorganic fertilizer.
Phytoremediation is a promising new technology that uses higher plants to enhance biodegradation. Nutrient availability is an important factor governing the success of phytoremediation and can be regulated through the addition of fertilizer. A greenhouse study was conducted to assess the importance of nitrogen and phosphorus for the phytoremediation of petroleum sludge. Degradation of total petroleum hydrocarbons (TPH) was quantified for six fertilization rates and three vegetation treatments: bermuda grass [Cynodon dactylon (L.) Pers.], tall fescue (Festuca arundinacea Schreb.), and an unvegetated control. During the first 6 mo of the experiment, TPH declined by an average of 49% with no significant differences between treatments. After 1 yr, TPH degradation was significantly greater in both vegetated treatments with a mean TPH reduction of 68% for bermuda, 62% for fescue, and 57% for the unvegetated control. Degradation of TPH in the fescue and bermuda treatments was significantly lower in the treatments in which no fertilizer was added or N and P were added simply to maintain plant growth compared with the higher rates of fertilization. For this short-term, greenhouse experiment, optimal remediation was obtained by fertilization that produced a C to N to P ratio of 100:2:0.2.


Racial discrimination and allostatic load among First Nations Australians: a nationally representative cross-sectional study.
Increased allostatic load is linked with racial discrimination exposure, providing a mechanism for the biological embedding of racism as a psychosocial stressor. We undertook an examination of how racial discrimination interacts with socioecological, environmental, and health conditions to affect multisystem dysregulation in a First Nations population. We conducted latent class analysis (LCA) using indicators of life stress, socioeconomic background, and physical and mental health from a nationally representative sample of Australian Aboriginal adults (N = 2056). We used LCA with distal outcomes to estimate the effect of the latent class variable on our derived allostatic load index and conducted a stratified analysis to test whether allostatic load varied based on exposure to racial discrimination across latent classes. Our psychosocial, environmental, and health measures informed a four-class structure; 'Low risk', 'Challenged but healthy', 'Mental health risk' and 'Multiple challenges'. Mean allostatic load was highest in 'Multiple challenges' compared to all other classes, both in those exposed (4.5; 95% CI: 3.9, 5.0) and not exposed (3.9; 95% CI: 3.7, 4.2) to racial discrimination. Allostatic load was significantly higher for those with exposure to racial discrimination in the 'Multiple challenges' class (t = 1.74, p = .04) and significantly lower in the 'Mental health risk' class (t = - 1.67, p = .05). Racial discrimination may not always modify physiological vulnerability to disease. Social and economic contexts must be considered when addressing the impact of racism, with a focus on individuals and sub-populations experiencing co-occurring life challenges.


[The "information test" in the diagnosis of ischemic heart disease].
The effect of psychophysiologic testing (the information test) on coronary flow was examined in 113 patients with coronary heart disease, 48 patients with neurocirculatory dystonia, 10 patients with essential hypertension and 15 normal subjects. Electrocardiographic evidence of coronary insufficiency was identified that can be found in 75.2% of coronary patients.


Pediatric Readmissions After Hospitalizations for Lower Respiratory Infections.
Lower respiratory infections (LRIs) are among the most common reasons for pediatric hospitalization and among the diagnoses with the highest number of readmissions. Characterizing LRI readmissions would help guide efforts to prevent them. We assessed variation in pediatric LRI readmission rates, risk factors for readmission, and readmission diagnoses. We analyzed 2008-2009 Medicaid Analytic eXtract data for patients <18 years of age in 26 states. We identified LRI hospitalizations based on a primary diagnosis of bronchiolitis, influenza, or community-acquired pneumonia or a secondary diagnosis of one of these LRIs plus a primary diagnosis of asthma, respiratory failure, or sepsis/bacteremia. Readmission rates were calculated as the proportion of hospitalizations followed by ≥1 unplanned readmission within 30 days. We used logistic regression with fixed effects for patient characteristics and a hospital random intercept to case-mix adjust rates and assess risk factors. Of 150 590 LRI hospitalizations, 8233 (5.5%) were followed by ≥1 readmission. The median adjusted hospital readmission rate was 5.2% (interquartile range: 5.1%-5.4%), and rates varied across hospitals (P < .0001). Infants (patients <1 year of age), boys, and children with chronic conditions were more likely to be readmitted. The most common primary diagnoses on readmission were LRIs (48.2%), asthma (10.0%), fluid/electrolyte disorders (3.4%), respiratory failure (3.3%), and upper respiratory infections (2.7%). LRI readmissions are common and vary across hospitals. Multiple risk factors are associated with readmission, indicating potential targets for strategies to reduce readmissions. Readmission diagnoses sometimes seem related to the original LRI.


Venous Thromboembolism Prophylaxis and Risk in the Inpatient and Outpatient Continuum of Care Among Hospitalized Acutely Ill Patients in the US: A Retrospective Analysis.
Venous thromboembolism (VTE) is a leading cause of preventable morbidity and mortality among hospitalized patients in the US. The objectives of this study were to examine VTE prophylaxis patterns and risk for VTE events during hospitalization and post-discharge among patients hospitalized for acute illnesses in the US. Acutely ill hospitalized patients were identified from the MarketScan databases (January 1, 2012-June 30, 2015). Proportions of patients that received inpatient and/or outpatient VTE prophylaxis were determined. VTE rates were calculated for the overall study population and for each subpopulation with each acute illness type. Risk for VTE events after the index admission was determined by Kaplan-Meier analysis. Of the acutely ill patients (n = 17,895, mean age: 58.4 years), most were hospitalized for infectious diseases (40.6%), followed by respiratory diseases (31.0%), cancer (10.7%), heart failure (10.4%), ischemic stroke (6.4%), and rheumatic diseases (0.9%). Among the entire study population, 59.1% did not receive any VTE prophylaxis, and only 7.1% received both inpatient and outpatient prophylaxis. Among the overall study population, cumulative VTE rate, including during index admission and within 6 months post-discharge, was 4.6%. VTE risk in the inpatient and outpatient continuum of care remained elevated up to 30-40 days after hospital admission, with 60.1% of VTEs occurring within 40 days of hospital admission. In this retrospective analysis of nearly 18,000 patients hospitalized for acute illnesses, 59.1% did not receive any VTE prophylaxis and only 7.1% received VTE prophylaxis in both the inpatient and outpatient continuum of care, despite significant VTE risk extending from hospitalization into the post-discharge period. Portola Pharmaceuticals.


Trapping of single nano-objects in dynamic temperature fields.
In this article we explore the dynamics of a Brownian particle in a feedback-free dynamic thermophoretic trap. The trap contains a focused laser beam heating a circular gold structure locally and creating a repulsive thermal potential for a Brownian particle. In order to confine a particle the heating beam is steered along the circumference of the gold structure leading to a non-trivial motion of the particle. We theoretically find a stability condition by switching to a rotating frame, where the laser beam is at rest. Particle trajectories and stable points are calculated as a function of the laser rotation frequency and are experimentally confirmed. Additionally, the effect of Brownian motion is considered. The present study complements the dynamic thermophoretic trapping with a theoretical basis and will enhance the applicability in micro- and nanofluidic devices.


zCall: a rare variant caller for array-based genotyping: genetics and population analysis.
zCall is a variant caller specifically designed for calling rare single-nucleotide polymorphisms from array-based technology. This caller is implemented as a post-processing step after a default calling algorithm has been applied. The algorithm uses the intensity profile of the common allele homozygote cluster to define the location of the other two genotype clusters. We demonstrate improved detection of rare alleles when applying zCall to samples that have both Illumina Infinium HumanExome BeadChip and exome sequencing data available. http://atguweb.mgh.harvard.edu/apps/zcall. firstname.lastname@example.com Supplementary data are available at Bioinformatics online.


Health-Related Quality of Life Outcomes in Patients with Myelodysplastic Syndromes with Ring Sideroblasts Treated with Luspatercept in the MEDALIST Phase 3 Trial.
Patients with myelodysplastic syndromes (MDS) often experience chronic anemia and long-term red blood cell transfusion dependence associated with significant burden on clinical and health-related quality of life (HRQoL) outcomes. In the MEDALIST trial (NCT02631070), luspatercept significantly reduced transfusion burden in patients with lower-risk MDS who had ring sideroblasts and were refractory to, intolerant to, or ineligible for prior treatment with erythropoiesis-stimulating agents. We evaluated the effect of luspatercept on HRQoL in patients enrolled in MEDALIST using the EORTC QLQ-C30 and the QOL-E questionnaire. Change in HRQoL was assessed every 6 weeks in patients receiving luspatercept with best supportive care (+ BSC) and placebo + BSC from baseline through week 25. No clinically meaningful within-group changes and between-group differences across all domains of the EORTC QLQ-C30 and QOL-E were observed. On one item of the QOL-E MDS-specific disturbances domain, patients treated with luspatercept reported marked improvements in their daily life owing to the reduced transfusion burden, relative to placebo. Taken together with previous reports of luspatercept + BSC reducing transfusion burden in patients from baseline through week 25 in MEDALIST, these results suggest luspatercept may offer a treatment option for patients that reduces transfusion burden while providing stability in HRQoL.


Genomic DNA-chip hybridization reveals a higher incidence of genomic amplifications in pancreatic cancer than conventional comparative genomic hybridization and leads to the identification of novel candidate genes.
Genomic analyses aimed at the detection of high-level DNA amplifications were performed on 13 widely used pancreatic cancer cell lines and 6 pancreatic tumor specimens. For these analyses, array-based comparative genomic hybridization (Matrix-CGH) onto dedicated microarrays was used. In comparison with chromosomal CGH (eight amplifications), a >3-fold number of DNA amplifications was detected (n = 29). The most frequent amplifications mapped to 7p12.3 (three pancreatic cancer cell lines and three pancreatic tumor specimens), 8q24 (four pancreatic cancer cell lines and one pancreatic tumor specimen), 11q13 (three pancreatic cancer cell lines and three pancreatic tumor specimens), and 20q13 (four pancreatic cancer cell lines and three pancreatic tumor specimens). Genes contained in the consensus regions were MYC (8q24), EGFR (7p12.3), and FGF3 (11q13). In six of seven pancreatic cancer cell lines and pancreatic tumor specimens with 20q13 amplifications, the novel candidate gene NFAT C2, which plays a role in the activation of cytokines, was amplified. Other amplifications also affected genes for which a pathogenetic role in pancreatic carcinoma has not been described, such as BCL10 and BCL6, two members of the BCL family. A subset of amplified genes was checked for overexpression by means of real-time PCR, revealing the highest expression levels for BCL6 and BCL10. Thus, Matrix-CGH allows the detection of a high number of amplifications, resulting in the identification of novel candidate genes in pancreatic cancer.


Critical analysis of the Council for International Organizations of Medical Sciences 2016 International Guidelines for health-related research involving humans.
This paper presents a preliminary discussion of the Council for International Organizations of Medical Sciences (CIOMS), recently issued "International Ethical Guidelines for Health-related Research Involving Humans" (2016) that acknowledges the document's declared concern of the protection of human subjects and awareness of their needs and interests in "low-resource settings". Nevertheless, guideline recommendations present exceptional situations –vulnerability, mental incompetence- wherein voluntary and consented participation may be reduced or omitted under three concurrent conditions: compelling scientific value, the need to include persons that will not benefit directly from participation, exposure to minimal or slight risks. CIOMS 2016 extends the range of issues than need deliberation and regulation, but it does not clarify controversial issues in research ethics. The indetermination of special situations that would allow incomplete or absent informed disclosure weakens rigorous ethical norms and opens up to slippery slopes of permissiveness that may be harmful to host-nations and participants of biomedical studies.


Echocardiography. The role of this noninvasive test in the geriatric population.
Echocardiography assumes a major role in the diagnosis and evaluation of cardiovascular disease in the older population. Available diagnostic modalities are briefly reviewed and their role in identifying disease entities common in a geriatric population is discussed. The prognostic implications of echocardiographic findings of mitral annular calcification and aortic sclerosis are reviewed, as are the use of echo and Doppler techniques in the assessment of congestive heart failure. Particular emphasis is placed on the evaluation of ischemic heart disease using stress echocardiography, the diagnosis and assessment of degenerative aortic valve disease, and the role of transesophageal echo in the management of atrial fibrillation and stroke.


Ligand substitution from the (eta5-DMP)Mn(CO)2(Solv) [DMP = 2,5-dimethylpyrrole, Solv = solvent] complexes: to ring slip or not to ring slip?
The mechanism and energetics of the displacement of solvent from photolytically generated (eta(5)-DMP)Mn(CO)(2)(Solv) complexes has been studied [DMP = 2,5-dimethylpyrrole, Solv = solvent]. Rate enhancement relative to the eta(5)-cyclopentadienyl (Cp) system is not observed in the displacement of weakly bound solvents. The bond dissociation enthalpies obtained from the kinetic analysis are in good agreement with the values obtained by detailed density functional theory (DFT) calculations. The results indicate that for both the Cp and the DMP based systems the displacement of weakly bound solvents proceeds by a dissociative or I(d) mechanism. This is in sharp contrast to CO displacement from (eta(5)-DMP)Mn(CO)(3), which is known to proceed by an associative mechanism by way of an eta(3) ring slip intermediate. The associative substitution pathway only becomes competitive with the dissociative channel when the Mn-Solv bond dissociation enthalpy is more than 33 kcal/mol.


Understanding barriers to mental health service utilization for adolescents in rural Australia.
There is a general paucity of research in the area of rural adolescent mental health in Australia, and in particular a lack of data regarding the experiences of rural adolescents who seek help for mental health problems. This study used a qualitative approach to data collection and analysis in order to assist understanding of the barriers to mental health service utilization for young people in rural communities. A series of interviews were conducted with each of the study's participants, who ranged in age from 15 to 17 years. All participants were clients of the Child and Adolescent Mental Health Services in the rural cities of Horsham and Ararat, Victoria, Australia. Participants described how the lack of reliable transport to and from the mental health service affected the utilization of the service by rural young people. They also expressed concern regarding a lack of qualified professionals in their region who specialize in child and adolescent mental health. Participants reported frustration at long waiting lists and the lack of an after-hours service. One participant shared her experiences of deliberate self-harm to in order to gain access. Results also revealed that rural gossip networks and social visibility within rural communities compounded the experience of stigma and social exclusion for these young people. Furthermore, participants explained how these experiences negatively impacted on their utilization of the mental health service and their progress towards recovery. There are several barriers to mental health service utilization for rural adolescents which affect both their decision to access help as well as their ability to engage effectively with mental health services over time. Clinicians who work with rural adolescents need to be mindful of the influence of rural culture on mental health service utilization by young people. The co-location of mental health services and general health services is suggested as one way to reduce the fear associated with 'being seen' entering a stand-alone mental health service. It is suggested that treatment programs for adolescents in rural areas address the different types of stigma that these young people are likely to encounter. Furthermore, community and school-based interventions aimed at reducing the social stigma of young people with mental illness in rural areas is recommended.


[Changes of mitochondria membrane potential of the uterine smooth muscle under Mg2+ and Ca2+ influence].
Using the method of flow cytometry and potential-sensitive fluorescent dye TMRM the effect of divalent cations on the membrane potential of isolated myometrium mitochondria was studied. It was shown, that Mg2+ (7 mM) addition to the incubation medium induced mitochondrial membrane hyperpolarization. In the case of protonophore CCCP (1 microM) preliminary presence in the incubation medium, Mg2+ addition did not lead to membrane potential hyperpolarization. Addition of Ca24+ leads to membrane potential dissipation. Ca2+ -induced mitochondrial depolarization is the same as NaN3 (10 mM)-induced depolarization. It also was shown that Mg2+ (7 mM) preliminary presence in the incubation medium did not protect mitochondria from Ca2+ -induced depolarization.


Directed evolution of mammalian cytochrome P450 2B1: mutations outside of the active site enhance the metabolism of several substrates, including the anticancer prodrugs cyclophosphamide and ifosfamide.
Cytochrome P450 2B1 has been subjected to directed evolution to investigate the role of amino acid residues outside of the active site and to engineer novel, more active P450 catalysts. A high throughput screening system was developed to measure H(2)O(2)-supported oxidation of the marker fluorogenic substrate 7-ethoxy-4-trifluoromethylcoumarin (7-EFC). Random mutagenesis by error-prone polymerase chain reaction and activity screening were optimized using the L209A mutant of P450 2B1 in an N-terminally modified construct with a C-terminal His tag (P450 2B1dH). Two rounds of mutagenesis and screening and one subcloning step yielded the P450 2B1 quadruple mutant V183L/F202L/L209A/S334P, which demonstrated a 6-fold higher k(cat) than L209A. Further random or site-directed mutagenesis did not improve the activity. When assayed in an NADPH-supported reconstituted system, V183L/L209A demonstrated lower 7-EFC oxidation than L209A. Therefore, F202L/L209A/S334P was generated, which showed a 2.5-fold higher k(cat)/K(m) for NADPH-dependent 7-EFC oxidation than L209A. F202L/L209A/S334P also showed enhanced catalytic efficiency with 7-benzyloxyresorufin, benzphetamine, and testosterone, and a 10-fold increase in stereoselectivity for testosterone 16alpha-versus 16beta-hydroxylation compared with 2B1dH. Enhanced catalytic efficiency of F202L/L209A/S334P was also retained in the full-length P450 2B1 background with 7-EFC and testosterone as substrates. Finally, the individual mutants were tested for metabolism of the anti-cancer prodrugs cyclophosphamide and ifosfamide. Several of the mutants showed increased metabolism via the therapeutically beneficial 4-hydroxylation pathway, with L209A/S334P showing 2.8-fold enhancement of k(cat)/K(m) with cyclophosphamide and V183L/L209A showing 3.5-fold enhancement with ifosfamide. Directed evolution can thus be used to enhance P450 2B1 catalytic efficiency across a panel of substrates and to identify functionally important residues distant from the active site.


Behavioral changes in the housing market before and after the Covid-19 lockdown.
We exploit unique Norwegian day-by-day transaction and hour-by-hour bidding logs data in order to examine how market participants reacted to the spreading news of Covid-19 in early March 2020, the lockdown on March 12, and the re-opening on April 20. We observe changes on the date of the lockdown in transaction volumes, sell-prediction spreads, exploitative bidding behavior, and seller confidence. However, when we compare observed price developments with our estimated counter-factual price developments, we find that about half of the total fall in prices had already occurred before the lockdown was implemented. The re-opening completely reverses the lockdown effect on prices. We show that voluntary behavioral changes, as well as lockdown and re-opening effects, are visible in various measures of social mobility, and that changes in daily news sentiment correlate with the abnormal price movements during this period.


Emotion regulation and mental health among professionals of long-term care institutions for older adults: the mediating role of work engagement.
This study examined whether work engagement mediated the association between emotion regulation and mental health of professionals working in long-term care institutions for older adults in Brazil. A cross-sectional study with a total of 104 professionals working in 13 long-term care facilities for the elderly in Brazil was conducted. Emotion regulation (expressive suppression and cognitive reappraisal), work engagement (vigor, dedication, and absorption), and mental health (depression, anxiety, and stress) were measured using self-report scales. Multiple mediation models were used to test the mediating role of engagement. Expressive suppression was associated with more vigor, which in turn was associated with less anxiety and stress; cognitive reappraisal was associated with more vigor, which in turn was associated with less anxiety and stress. No indirect effects were found for depression. Expressive suppression and cognitive reappraisal strategies to regulate emotions seem to be associated with more vigor in terms of resilience and persistence to deal with work demands in these professionals. These results seem to suggest that workers who are more capable of regulating their emotions (using both expressive suppression and cognitive reappraisal in a flexible way according to work demands) are more likely to be resilient, invest in their profession, and be persistent in the face of difficulties. This, in turn, contributes to protecting them from experiencing anxiety and stress.


Predicting the Future: Using Simulation Modeling to Forecast Patient Flow on General Medicine Units.
Hospitals are complex adaptive systems within which multiple components such as patients, practitioners, facilities, and technology interact. A careful approach to optimization of this complex system is needed because any change can result in unexpected deleterious effects. One such approach is discrete event simulation, in which what-if scenarios allow researchers to predict the impact of a proposed change on the system. However, studies illustrating the application of simulation in optimization of general internal medicine (GIM) team inpatient operations are lacking. Administrative data about admissions and discharges, data from a time-motion study, and expert opinion on workflow were used to construct the simulation model. Then, the impact of four changes: aligning medical teams with nursing units, adding a hospitalist team, adding a nursing unit, and adding both a nursing unit and hospitalist team with higher admission volume were modeled on key hospital operational metrics. Aligning medical teams with nursing units improved team metrics for aligned teams but shifted patients to unaligned teams. Adding a hospitalist team had little benefit, but adding a nursing unit improved system metrics. Both adding a hospitalist team and a nursing unit would be required to maintain operational metrics with increased patient volume. Using simulation modeling, we provided data on the implications of four possible strategic changes on GIM inpatient units, providers, and patient throughput. Such analyses may be a worthwhile investment to study strategic decisions and make better choices with fewer unintended consequences.


Roles of autolysin and pneumolysin in middle ear inflammation caused by a type 3 Streptococcus pneumoniae strain in the chinchilla otitis media model.
Streptococcus pneumoniae cell wall and pneumolysin are important contributors to pneumococcal pathogenicity in some animal models. To further explore these factors in middle ear inflammation caused by pneumococci, penicillin-induced inflammatory acceleration was studied by using three closely related pneumococcal strains: a wild-type 3 strain (WT3), its pneumolysin-negative derivative (P-1), and into autolysin-negative derivative (A-1). Both middle ears of chinchillas were inoculated with one of the three pneumococcal strains. During the first 12 h, all three strains grew in vivo at the same rate, and all three strains induced similar inflammatory cell responses in middle ear fluid (MEF). Procaine penicillin G was given as 12 h to one-half of the animals in each group, and all treated chinchillas had sterile MEF at 24 h. Penicillin significantly accelerated MEF inflammatory cell influx into WT3-and P-1-infected ears at 18 and 24 h in comparison with the rate for penicillin-treated A-1-infected ears. Inflammatory cell influx was slightly, but not significantly, greater after treatment of WT3 infection than after treatment of P-1 infection. Interleukin (IL)-1beta and IL-6, but not IL-8, concentrations in MEF at 24 h reflected the penicillin effect on MEF inflammatory cells; however, differences between treatment groups were not significant. Results suggest that pneumococcal otitis media pathogenesis is triggered principally by the inflammatory effects of intact and lytic cell wall products in the middle ear, with at most a modes additional pneumolysin effect. Investigation strategies that limit the release of these products or neutralize them warrant further investigation.


Improving service quality in primary care.
A framework for improving health care service quality was implemented at a 12-provider family medicine practice in 2010. A national patient satisfaction research vendor conducted weekly telephone surveys of 840 patients served by that practice: 280 patients served in 2009, and 560 served during 2010 and 2011. After the framework was implemented, the proportion of "excellent" ratings of provider service (the highest rating on a 5-point scale) increased by 5% to 9%, most notably thoroughness (P = .04), listening (P = .04), and explaining (P = .04). Other improvements included prompt test result notification and telephone staff courtesy (each by 10%, P = .02), as well as teamwork (by 8%, P = .04). Overall quality increased by 10% (P = .01), moving the practice from the 68th to the 91st percentile of medical practices in the research vendor's database. Improvements in patient satisfaction suggest that this framework may be useful in value-based payment models.


Post-Exposure Anti-Ricin Treatment Protects Swine Against Lethal Systemic and Pulmonary Exposures.
Ricin, a plant-derived toxin originating from the seeds of Ricinus communis (castor bean plant), is one of the most lethal toxins known. To date, there is no approved post-exposure therapy for ricin exposures. This work demonstrates for the first time the therapeutic efficacy of equine-derived anti-ricin F(ab')2 antibodies against lethal pulmonary and systemic ricin exposures in swine. While administration of the antitoxin at 18 h post-exposure protected more than 80% of both intratracheally and intramuscularly ricin-intoxicated swine, treatment at 24 h post-exposure protected 58% of the intramuscular-exposed swine, as opposed to 26% of the intratracheally exposed animals. Quantitation of the anti-ricin neutralizing units in the anti-toxin preparations confirmed that the disparate protection conferred to swine subjected to the two routes of exposure stems from variance between the two models. Furthermore, dose response experiments showed that approximately 3 times lesser amounts of antibody are needed for high-level protection of the intramuscularly compared to the intratracheally intoxicated swine. This study, which demonstrates the high-level post-exposure efficacy of anti-ricin antitoxin at clinically relevant time-points in a large animal model, can serve as the basis for the formulation of post-exposure countermeasures against ricin poisoning in humans.


In vivo Three-Dimensional Superresolution Fluorescence Tracking using a Double-Helix Point Spread Function.
The point spread function (PSF) of a widefield fluorescence microscope is not suitable for three-dimensional super-resolution imaging. We characterize the localization precision of a unique method for 3D superresolution imaging featuring a double-helix point spread function (DH-PSF). The DH-PSF is designed to have two lobes that rotate about their midpoint in any transverse plane as a function of the axial position of the emitter. In effect, the PSF appears as a double helix in three dimensions. By comparing the Cramer-Rao bound of the DH-PSF with the standard PSF as a function of the axial position, we show that the DH-PSF has a higher and more uniform localization precision than the standard PSF throughout a 2 μm depth of field. Comparisons between the DH-PSF and other methods for 3D super-resolution are briefly discussed. We also illustrate the applicability of the DH-PSF for imaging weak emitters in biological systems by tracking the movement of quantum dots in glycerol and in live cells.