Bone marrow-derived fibroblast precursors mediate ischemic cardiomyopathy in mice.
We previously described a mouse model of fibrotic ischemia/reperfusion cardiomyopathy (I/RC) arising from daily, brief coronary occlusion. One characteristic of I/RC was the prolonged elevation of monocyte chemoattractant protein 1 (MCP-1), which was obligate to its phenotype and may contribute to the uptake of bloodborne cells. Here we describe in I/RC hearts a population of small spindle-shaped fibroblasts that were highly proliferative and expressed collagen I and alpha-smooth muscle actin (myofibroblast markers), CD34 (a precursor marker), and CD45 (a hematopoietic marker). These cells represented 3% of all nonmyocyte live cells. To confirm the cells' bone marrow origin, chimeric mice were created by the rescue of irradiated C57BL/6 mice with marrow from ROSA26, a congenic line expressing lacZ. I/RC resulted in a large population of spindle-shaped fibroblasts containing lacZ. We postulated that the fibroblast precursors represented a developmental path for a subset of monocytes, whose phenotype we have shown to be influenced by serum amyloid P (SAP). Thus, we administered SAP in vivo, which markedly reduced the number of proliferative spindle-shaped fibroblasts and completely prevented I/RC-induced fibrosis and global ventricular dysfunction. By contrast, SAP did not suppress the inflammation or chemokine expression seen in I/RC. SAP, a member of the pentraxin family, binds to Fcgamma receptors and modifies the pathophysiological function of monocytes. Our data suggest that SAP interferes with assumption of a fibroblast phenotype in a subset of monocytes and that SAP may be an important regulator in the linkage between inflammation and nonadaptive fibrosis in the heart.


The binding of vimentin to human erythrocyte membranes: a model system for the study of intermediate filament-membrane interactions.
We have characterized the association of the intermediate filament protein, vimentin, with the plasma membrane, using radioiodinated lens vimentin and various preparations of human erythrocyte membrane vesicles. Inside-out membrane vesicles (IOVs), depleted of spectrin and actin, bind I125-vimentin in a saturable manner unlike resealed, right-side-out membranes which bind negligible amounts of vimentin in an unsaturable fashion. The binding of vimentin to IOVs is abolished by trypsin or acid treatment of the vesicles. Extraction of protein 4.1 or reconstitution of the membranes with purified spectrin do not basically affect the association. However, removal of ankyrin (band 2.1) significantly lowers the binding. Upon reconstitution of depleted vesicles with purified ankyrin, the vimentin binding function is restored. If ankyrin is added in excess the binding of vimentin to IOVs is quantitatively inhibited, whereas protein 4.1, the cytoplasmic fragment of band 3, band 6, band 4.5 (catalase), or bovine serum albumin do not influence it. Preincubation of the IOVs with a polyclonal anti-ankyrin antibody blocks 90% of the binding. Preimmune sera and antibodies against spectrin, protein 4.1, glycophorin A, and band 3 exhibit no effect. On the basis of these data, we propose that vimentin is able to associate specifically with the erythrocyte membrane skeleton and that ankyrin constitutes its major attachment site.


Pulmonary fibrotic response to aspiration of multi-walled carbon nanotubes.
Multi-walled carbon nanotubes (MWCNTs) are new manufactured nanomaterials with a wide spectrum of commercial applications. To address the hypothesis that MWCNTs cause persistent pulmonary pathology, C57BL/6J mice were exposed by pharyngeal aspiration to 10, 20, 40 or 80 μg of MWCNTs (mean dimensions of 3.9 μm × 49 nm) or vehicle. Lungs were preserved at 1, 7, 28 and 56 days post- exposure to determine the potential regions and target cells for impact by MWCNT lung burden. Morphometric measurement of Sirius Red staining was used to assess the connective tissue response. At 56 days post-exposure, 68.7 ± 3.9, 7.5 ± 1.9 and 22.0 ± 5.1 percent (mean ± SE, N = 8) of the MWCNT lung burden were in alveolar macrophages, alveolar tissue and granulomatous lesions, respectively. The subpleural tissues contained 1.6% of the MWCNT lung burden. No MWCNTs were found in the airways at 7, 28 or 56 days after aspiration The connective tissue in the alveolar interstitium demonstrated a progressive increase in thickness over time in the 80 μg exposure group (0.12 ± 0.01, 0.12 ± 0.01, 0.16 ± 0.01 and 0.19 ± 0.01 μm for 1, 7, 28 and 56 days post-exposure (mean ± SE, N = 8)). Dose-response determined at 56 days post-exposure for the average thickness of connective tissue in alveolar septa was 0.11 ± 0.01, 0.14 ± .02, 0.14 ± 0.01, 0.16 ± 0.01 and 0.19 ± 0.01 μm (mean ± SE, N = 8) for vehicle, 10, 20, 40 and 80 μg dose groups, respectively. The distribution of lung burden was predominately within alveolar macrophages with approximately 8% delivery to the alveolar septa, and a smaller but potentially significant burden to the subpleural tissues. Despite the relatively low fraction of the lung burden being delivered to the alveolar tissue, the average thickness of connective tissue in the alveolar septa was increased over vehicle control by 45% in the 40 μg and 73% in the 80 μg exposure groups. The results demonstrate that MWCNTs have the potential to produce a progressive, fibrotic response in the alveolar tissues of the lungs. However, the increases in connective tissue per μg dose of MWCNTs to the interstitium are significantly less than those previously found for single-walled carbon nanotubes (SWCNTs).


In vitro motility of liver connexin vesicles along microtubules utilizes kinesin motors.
Trafficking of the proteins that form gap junctions (connexins) from the site of synthesis to the junctional domain appears to require cytoskeletal delivery mechanisms. Although many cell types exhibit specific delivery of connexins to polarized cell sites, such as connexin32 (Cx32) gap junctions specifically localized to basolateral membrane domains of hepatocytes, the precise roles of actin- and tubulin-based systems remain unclear. We have observed fluorescently tagged Cx32 trafficking linearly at speeds averaging 0.25 μm/s in a polarized hepatocyte cell line (WIF-B9), which is abolished by 50 μM of the microtubule-disrupting agent nocodazole. To explore the involvement of cytoskeletal components in the delivery of connexins, we have used a preparation of isolated Cx32-containing vesicles from rat hepatocytes and assayed their ATP-driven motility along stabilized rhodamine-labeled microtubules in vitro. These assays revealed the presence of Cx32 and kinesin motor proteins in the same vesicles. The addition of 50 μM ATP stimulated vesicle motility along linear microtubule tracks with velocities of 0.4-0.5 μm/s, which was inhibited with 1 mM of the kinesin inhibitor AMP-PNP (adenylyl-imidodiphosphate) and by anti-kinesin antibody but only minimally affected by 5 μM vanadate, a dynein inhibitor, or by anti-dynein antibody. These studies provide evidence that Cx32 can be transported intracellularly along microtubules and presumably to junctional domains in cells and highlight an important role of kinesin motor proteins in microtubule-dependent motility of Cx32.


Molecular mechanisms of lipopolysaccharide-induced cyclooxygenase-2 expression in human neutrophils: involvement of the mitogen-activated protein kinase pathway and regulation by anti-inflammatory cytokines.
Neutrophils are an important cellular source of proinflammatory mediators, whose regulation may be of potential benefit for the treatment of a number of inflammatory diseases. However, the mechanisms of lipopolysaccharide (LPS)-induced neutrophil activation and its regulation by anti-inflammatory cytokines have not yet been fully elucidated. Recent studies have revealed that mitogen-activated protein kinases (MAPK) play a crucial role in the generation of proinflammatory mediators in some cell types. Therefore, we conducted this study to determine whether MAPK activation could be involved in prostaglandin E(2) (PGE(2)) production and cyclooxygenase (COX)-2 expression in LPS-stimulated human neutrophils. PD98059 (MEK1 inhibitor) and SB203580 (p38(MAPK) inhibitor) reduced PGE(2) production as well as COX-2 expression in LPS-stimulated neutrophils. In addition, both extracellular signal-regulated protein kinase (ERK) and p38(MAPK) were phosphorylated and activated in time- and dose-dependent manners. Since we previously showed that IL-10 and IL-4 similarly inhibited COX-2 expression in LPS-stimulated neutrophils, we next tested the effects of IL-10 and IL-4 on the phosphorylation and activation of both kinases. IL-10 inhibited the phosphorylation and activation of p38(MAPK), but not ERK. In addition, IL-4 caused a marginal inhibition in the activation of p38(MAPK). Taken together, these results suggest that both ERK and p38(MAPK) pathways are involved in LPS-induced COX-2 expression and PGE(2) production in neutrophils, and IL-10 and IL-4 inhibit neutrophil prostanoid synthesis by down-regulating the activation of p38(MAPK).


Lactose malabsorption and intolerance: pathogenesis, diagnosis and treatment.
Lactose malabsorption is a common condition caused by reduced expression or activity of lactase in the small intestine. In such patients, lactose intolerance is characterized by abdominal symptoms (e.g. nausea, bloating, and pain) after ingestion of dairy products. The genetic basis of lactose malabsorption is established and several tests for this condition are available, including genetic, endoscopic, and H2-breath tests. In contrast, lactose intolerance is less well understood. Recent studies show that the risk of symptoms after lactose ingestion depends on the dose of lactose, lactase expression, intestinal flora, and sensitivity of the gastrointestinal tract. Lactose intolerance has recently been defined as symptoms developing after ingestion of lactose which do not develop after placebo challenge in a person with lactose maldigestion. Such blinded testing might be especially important in those with functional gastrointestinal diseases in whom self-reported lactose intolerance is common. However, placebo-controlled testing is not part of current clinical practice. Updated protocols and high-quality outcome studies are needed. Treatment options of lactose intolerance include lactose-reduced diet and enzyme replacement. Documenting the response to multiple doses can guide rational dietary management; however, the clinical utility of this strategy has not been tested. This review summarizes the genetic basis, diagnosis, and treatment of lactose malabsorption and intolerance.


Investigation of the effect of ginger on the lipid levels. A double blind controlled clinical trial.
To study the effect of fine powder of ginger on lipid level in volunteer patients. This is a double blind controlled clinical trial study in 2 cardiac clinics Cardiac Disease Clinic, Babol, north of Iran, between April to May 2004. We randomly divided the patients with hyperlipidemia into 2 groups, treatment group (receiving ginger capsules 3 g/day in 3 divided doses) and placebo group (lactose capsule 3 g/day in 3 divided doses) for 45 days. All subjects with diabetes mellitus, hypothyroidism, nephrotic syndrome, and alcohol drinking, pregnancy and peptic ulcer were excluded. Lipid concentrations profile before and after treatment was measured by enzymatic assay. Forty-five patients in the treatment group and 40 patients in placebo group participated in this study. There was a significant reduce in triglyceride, cholesterol, low density lipoprotein (LDL), very low density lipoprotein (VLDL), levels of before and after study separately in each group (p<0.05). Mean changes in triglyceride and cholesterol levels of ginger group were significantly higher than placebo group (p<0.05). Mean reduction in LDL level and increase in high density lipoprotein level of ginger group were higher than the placebo group, but in VLDL level of placebo was higher than ginger (p>0.05). The results show that ginger has a significant lipid lowering effect compared to placebo.


Work attitudes of older RNs.
Using data collected from 1,906 RNs from Metropolitan Statistical Areas in 29 states, the purpose of this study was to examine the characteristics and work attitudes of older RNs compared to RNs less than age 50 at two time periods, and compare among the older RNs those who are working in nursing, working outside nursing, and not working. Older RNs reported more distributive justice (fairness of rewards), work group cohesion, and supervisory support and less organizational constraint, and quantitative workload than younger RNs. Overall, older RNs were more satisfied, had greater organizational commitment, and had less desire to quit than younger RNs. There were no significant differences between older and younger RNs for autonomy, mentor support, or variety. Strategic efforts by employers and government could be used to retain older workers, attract RNs working in nonnursing settings back into nursing, and recruit retired RNs into the nursing workforce.


Spin current generation in organic antiferromagnets.
Spin current-a flow of electron spins without a charge current-is an ideal information carrier free from Joule heating for electronic devices. The celebrated spin Hall effect, which arises from the relativistic spin-orbit coupling, enables us to generate and detect spin currents in inorganic materials and semiconductors, taking advantage of their constituent heavy atoms. In contrast, organic materials consisting of molecules with light elements have been believed to be unsuited for spin current generation. Here we show that a class of organic antiferromagnets with checker-plate type molecular arrangements can serve as a spin current generator by applying a thermal gradient or an electric field, even with vanishing spin-orbit coupling. Our findings provide another route to create a spin current distinct from the conventional spin Hall effect and open a new field of spintronics based on organic magnets having advantages of small spin scattering and long lifetime.


Scaffold-based discovery of indeglitazar, a PPAR pan-active anti-diabetic agent.
In a search for more effective anti-diabetic treatment, we used a process coupling low-affinity biochemical screening with high-throughput co-crystallography in the design of a series of compounds that selectively modulate the activities of all three peroxisome proliferator-activated receptors (PPARs), PPARalpha, PPARgamma, and PPARdelta. Transcriptional transactivation assays were used to select compounds from this chemical series with a bias toward partial agonism toward PPARgamma, to circumvent the clinically observed side effects of full PPARgamma agonists. Co-crystallographic characterization of the lead molecule, indeglitazar, in complex with each of the 3 PPARs revealed the structural basis for its PPAR pan-activity and its partial agonistic response toward PPARgamma. Compared with full PPARgamma-agonists, indeglitazar is less potent in promoting adipocyte differentiation and only partially effective in stimulating adiponectin gene expression. Evaluation of the compound in vivo confirmed the reduced adiponectin response in animal models of obesity and diabetes while revealing strong beneficial effects on glucose, triglycerides, cholesterol, body weight, and other metabolic parameters. Indeglitazar has now progressed to Phase II clinical evaluations for Type 2 diabetes mellitus (T2DM).


Unveiling astrocytic control of cerebral blood flow with optogenetics.
Cortical neural activities lead to changes in the cerebral blood flow (CBF), which involves astrocytic control of cerebrovascular tone. However, the manner in which astrocytic activity specifically leads to vasodilation or vasoconstriction is difficult to determine. Here, cortical astrocytes genetically expressing a light-sensitive cation channel, channelrhodopsin-2 (ChR2), were transcranially activated with a blue laser while the spatiotemporal changes in CBF were noninvasively monitored with laser speckle flowgraphy in the anesthetised mouse cortex. A brief photostimulation induced a fast transient increase in CBF. The average response onset time was 0.7 ± 0.7 sec at the activation foci, and this CBF increase spread widely from the irradiation spot with an apparent propagation speed of 0.8-1.1 mm/sec. The broad increase in the CBF could be due to a propagation of diffusible vasoactive signals derived from the stimulated astrocytes. Pharmacological manipulation showed that topical administration of a K(+) channel inhibitor (BaCl2; 0.1-0.5 mM) significantly reduced the photostimulation-induced CBF responses, which indicates that the ChR2-evoked astrocytic activity involves K(+) signalling to the vascular smooth muscle cells. These findings demonstrate a unique model for exploring the role of the astrocytes in gliovascular coupling using non-invasive, time-controlled, cell-type specific perturbations.


Alpha-fetoprotein and prognosis in acute liver failure.
Serum concentrations of alpha-fetoprotein (AFP), variably elevated during liver injury, have been suggested to be of prognostic importance in acute liver failure (ALF), higher values being associated with improved outcome. Using a nephelometric assay, we measured AFP in sera obtained on admission from 206 patients prospectively enrolled in the US ALF Study, and on day 3 in 162 of these patients. The AFP ratio was defined as the day 3 AFP concentration divided by that observed on day 1. Median (range) admission serum AFP in all patients was 8.1 (1-1,811) ng/mL and increased to 17.6 (1.1-1,162) ng/mL on day 3 (P < 0.001). Higher absolute levels were not associated with improved outcome. In fact, admission AFP levels were lower in survivors not receiving transplants than in those who died or were transplanted (P < 0.001), whereas there was no difference between the 2 groups on day 3 (P = 0.34). However, a rise in AFP values between day 1 and day 3 indicated a better prognosis: the AFP ratio was 2.2 (0.11-22.1) in spontaneous survivors and 0.87 (0.11-16.4) in nonsurvivors (P < 0.001). An increasing AFP level indicated by an AFP ratio >or=1 was observed in 70 of 98 (71%) survivors, whereas a ratio <1 was observed in 51 of 64 (80%) nonsurvivors. In conclusion, AFP values change dynamically during ALF. In this large prospective study, higher absolute values of AFP did not predict a favorable outcome, but a rising level of AFP over the first 3 hospital days frequently indicated survival.


Rationale and study design of the MyHEART study: A young adult hypertension self-management randomized controlled trial.
Young adults (18-39 year-olds) with hypertension have a higher lifetime risk for cardiovascular disease. However, less than 50% of young adults achieve hypertension control in the United States. Hypertension self-management programs are recommended to improve control, but have been targeted to middle-aged and older populations. Young adults need hypertension self-management programs (i.e., home blood pressure monitoring and lifestyle modifications) tailored to their unique needs to lower blood pressure and reduce the risks and medication burden they may face over a lifetime. To address the unmet need in hypertensive care for young adults, we developed MyHEART (My Hypertension Education And Reaching Target), a multi-component, theoretically-based intervention designed to achieve self-management among young adults with uncontrolled hypertension. MyHEART is a patient-centered program, based upon the Self-Determination Theory, that uses evidence-based health behavior approaches to lower blood pressure. Therefore, the objective of this study is to evaluate MyHEART's impact on changes in systolic and diastolic blood pressure compared to usual care after 6 and 12 months in 310 geographically and racially/ethnically diverse young adults with uncontrolled hypertension. Secondary outcomes include MyHEART's impact on behavioral outcomes at 6 and 12 months, compared to usual clinical care (increased physical activity, decreased sodium intake) and to examine whether MyHEART's effects on self-management behavior are mediated through variables of perceived competence, autonomy, motivation, and activation (mediation outcomes). MyHEART is one of the first multicenter, randomized controlled hypertension trials tailored to young adults with primary care. The design and methodology will maximize the generalizability of this study. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT03158051.


Delayed administration of a small molecule tropomyosin-related kinase B ligand promotes recovery after hypoxic-ischemic stroke.
Stroke is the leading cause of long-term disability in the United States, yet no drugs are available that are proven to improve recovery. Brain-derived neurotrophic factor stimulates neurogenesis and plasticity, processes that are implicated in stroke recovery. It binds to both the tropomyosin-related kinase B and p75 neurotrophin receptors. However, brain-derived neurotrophic factor is not a feasible therapeutic agent, and no small molecule exists that can reproduce its binding to both receptors. We tested the hypothesis that a small molecule (LM22A-4) that selectively targets tropomyosin-related kinase B would promote neurogenesis and functional recovery after stroke. Four-month-old mice were trained on motor tasks before stroke. After stroke, functional test results were used to randomize mice into 2 equally, and severely, impaired groups. Beginning 3 days after stroke, mice received LM22A-4 or saline vehicle daily for 10 weeks. LM22A-4 treatment significantly improved limb swing speed and accelerated the return to normal gait accuracy after stroke. LM22A-4 treatment also doubled both the number of new mature neurons and immature neurons adjacent to the stroke. Drug-induced differences were not observed in angiogenesis, dendritic arborization, axonal sprouting, glial scar formation, or neuroinflammation. A small molecule agonist of tropomyosin-related kinase B improves functional recovery from stroke and increases neurogenesis when administered beginning 3 days after stroke. These findings provide proof-of-concept that targeting of tropomyosin-related kinase B alone is capable of promoting one or more mechanisms relevant to stroke recovery. LM22A-4 or its derivatives might therefore serve as "pro-recovery" therapeutic agents for stroke.


Potential correlation between statins and pulp chamber calcification.
3-Hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins) are the first-line pharmaceuticals for the prevention and treatment of dyslipidemia. A recent investigation has shown that statins induced odontoblastic differentiation of dental pulp stem cells. Statins enhance the differentiation of human dental pulp cells by up-regulating mineralization nodules and odontogenic markers. This study tested the hypothesis that the systemic administration of statins results in increased dental pulp calcification. This retrospective case-control study used digital bitewing radiographs of mandibular molars. Subjects (N = 90) aged ≥60 years were assigned to either test (n = 45) or control (n = 45) groups based on the systemic use of statins. The dimensions of the pulp chambers were measured using a standardized method for height and mesiodistal distances. The chi-square test was used to analyze the data. Multiple linear regression model analysis was performed to explore the association between statin intake and pulp calcification. Three of the 45 mandibular molars in the test group exhibited almost complete pulp chamber obliteration. There was a significant reduction in pulp chamber height ratio shown in the statin group compared with the control group (P < .0001). When the mesiodistal width was compared between the 2 groups, there was no significant difference (P = .3730). The significant increase of calcification and loss of vertical height of the pulp chamber observed in mandibular molars in patients on statin medication indicated a possible increased odontoblastic activity. Therefore, systemic statins could be a contributing factor for pulp chamber calcification.


Polymicrobial endophthalmitis: prevalence, causative organisms, and visual outcomes.
The purpose of the present study is to evaluate the prevalence, causative organisms, and visual acuity outcome in patients with culture-proven polymicrobial endophthalmitis. The method used in this study is the non-comparative, consecutive case series using a retrospective analysis of patients diagnosed with polymicrobial endophthalmitis for the period 2000 to 2010. Polymicrobial endophthalmitis was identified in 43/1,107 (3.88%) patients. Forty-two patients had two isolates, and one patient had grown three isolates, yielding a total of 87 isolates. Gram-positive cocci were the most common isolate (n = 53; 60.9%) including Staphylococcus epidermidis (n = 14/53; 16.1%) and Streptococcus pneumoniae (n = 13/53; 13.8%). The etiologies included posttraumatic (n = 31/43; 72.1%) and postoperative (n = 9/43; 20.9%) endophthalmitis. Antibiotic susceptibilities among Gram-positive bacteria were vancomycin (100%) and chloramphenicol (96%). Susceptibilities among Gram-negative bacteria were ciprofloxacin (86.4%) and ofloxacin (81.2%). A maximum number of secondary interventions were done in traumatic cases (38.7%) and cases having coinfection with Gram-negative bacteria and fungus (66.7%). Visual acuity (VA) < 20/200 was more frequently observed in posttraumatic cases (n = 27/31; 87.1%) as compared with postoperative cases (n = 4/9; 44.4%). Of the 43 patients, only 9 patients (20.9%) achieved a VA ≥ 20/200 on final follow-up. Four out of twelve patients (33.3%), with fungus as one of the isolates, had a VA ≥ 20/200. Although polymicrobial infection in endophthalmitis is uncommon, it is generally associated with poor visual acuity outcomes especially in eyes with open-globe injuries. Coinfection with Gram-negative bacteria or fungi was associated with most unfavorable visual outcome.


Molecular imaging of bone marrow mononuclear cell survival and homing in murine peripheral artery disease.
This study aims to provide insight into cellular kinetics using molecular imaging after different transplantation methods of bone marrow-derived mononuclear cells (MNCs) in a mouse model of peripheral artery disease (PAD). MNC therapy is a promising treatment for PAD. Although clinical translation has already been established, there is a lack of knowledge about cell behavior after transplantation and about the mechanism whereby MNC therapy might ameliorate complaints of PAD. MNCs were isolated from F6 transgenic mice (FVB background) that express firefly luciferase (Fluc) and green fluorescence protein (GFP). Male FVB and C57Bl6 mice (n = 50) underwent femoral artery ligation and were randomized into 4 groups receiving the following: 1) single intramuscular (IM) injection of 2 × 10(6) MNCs; 2) 4 weekly IM injections of 5 × 10(5) MNCs; 3) 2 × 10(6) MNCs intravenously; and 4) phosphate-buffered saline as control. Cells were characterized by flow cytometry and in vitro bioluminescence imaging (BLI). Cell survival, proliferation, and migration were monitored by in vivo BLI, which was validated by ex vivo BLI, post-mortem immunohistochemistry, and flow cytometry. Paw perfusion and neovascularization was measured with laser Doppler perfusion imaging (LDPI) and histology, respectively. In vivo BLI revealed near-complete donor cell death 4 weeks after IM transplantation. After intravenous transplantation, BLI revealed that cells migrated to the injured area in the limb, as well as to the liver, spleen, and bone marrow. Ex vivo BLI showed presence of MNCs in the scar tissue and adductor muscle. However, no significant effects on neovascularization were observed, as monitored by LDPI and histology. This is one of the first studies to assess kinetics of transplanted MNCs in PAD using in vivo molecular imaging. MNC survival is short-lived, MNCs do not preferentially home to injured areas, and MNCs do not significantly stimulate perfusion in this particular model.


Calibrating a tuning fork for use as a scanning probe microscope force sensor.
Quartz tuning forks mounted with sharp tips provide an alternate method to silicon microcantilevers for probing the tip-substrate interaction in scanning probe microscopy. The high quality factor and stable resonant frequency of the tuning fork allow accurate measurements of small shifts in the resonant frequency as the tip approaches the substrate. To permit an accurate measure of surface interaction forces, the electrical and piezoelectromechanical properties of a tuning fork have been characterized using a fiber optical interferometer.


Clinical, epidemiologic, histopathologic and molecular features of an unexplained dermopathy.
Morgellons is a poorly characterized constellation of symptoms, with the primary manifestations involving the skin. We conducted an investigation of this unexplained dermopathy to characterize the clinical and epidemiologic features and explore potential etiologies. A descriptive study was conducted among persons at least 13 years of age and enrolled in Kaiser Permanente Northern California (KPNC) during 2006-2008. A case was defined as the self-reported emergence of fibers or materials from the skin accompanied by skin lesions and/or disturbing skin sensations. We collected detailed epidemiologic data, performed clinical evaluations and geospatial analyses and analyzed materials collected from participants' skin. We identified 115 case-patients. The prevalence was 3.65 (95% CI = 2.98, 4.40) cases per 100,000 enrollees. There was no clustering of cases within the 13-county KPNC catchment area (p = .113). Case-patients had a median age of 52 years (range: 17-93) and were primarily female (77%) and Caucasian (77%). Multi-system complaints were common; 70% reported chronic fatigue and 54% rated their overall health as fair or poor with mean Physical Component Scores and Mental Component Scores of 36.63 (SD = 12.9) and 35.45 (SD = 12.89), respectively. Cognitive deficits were detected in 59% of case-patients and 63% had evidence of clinically significant somatic complaints; 50% had drugs detected in hair samples and 78% reported exposure to solvents. Solar elastosis was the most common histopathologic abnormality (51% of biopsies); skin lesions were most consistent with arthropod bites or chronic excoriations. No parasites or mycobacteria were detected. Most materials collected from participants' skin were composed of cellulose, likely of cotton origin. This unexplained dermopathy was rare among this population of Northern California residents, but associated with significantly reduced health-related quality of life. No common underlying medical condition or infectious source was identified, similar to more commonly recognized conditions such as delusional infestation.


[Lipid profile and risk factors for cardiovascular diseases in medicine students].
To analyze the lipid profile and its correlation with risk factors for cardiovascular diseases (CVD) among medicine students. We assessed 153 students, regardless of sex, with age between 18 and 31 years old, submitted to lipid profile analysis, including serum levels of total cholesterol (TC), fraction of cholesterol of low (LDLc), high (HDLc) and very low density (VLDLc) lipoproteins and triglycerides (TG), besides of life habits and anthropometric data. Statistic analysis was employed, including the test of Mann Whitney, chi-square test, Pearson's correlation and multivariate analysis, by adopting a significance level for a value of p<0.05. Sedentary lifestyle (43.1%) and familial history for CVD, particularly hypertension (74.5%), were distinguished. The lipid profile showed desirable, although altered levels of TC, LDLc and TG were detected in 11.8%, 9.8% and 8.5% from the students, respectively, and reduced levels of HDLc in 12.4% of them. Women showed significantly reduced values for LDLc and high values for HDLc compared to men's (p=0.031 and p<0.0001, respectively). There was a significant association between lipid profile and, preferably, body mass index (BMI), sedentary lifestyle, ingestion of alcohol, contraceptive use, familial antecedents of cerebrovascular accident and dyslipidemia. Familial history for CVD, sedentary lifestyle and contraceptive use among medicine students showed frequent and associated to the lipid profile, as well as ingestion of alcohol and BMI. Although with desirable lipid profile, regardless of sex, higher levels of LDLc and reduced levels of HDLc in male sex provide a disadvantage to men compared to women.


Detection of inspiratory resistive loads after anaesthesia for minor surgery.
We measured the ability to detect inspiratory resistances in 22 patients undergoing minor gynaecological surgery, using linear resistances of 1.2-24.4 cm H2O litre-1 s, and ranked 1-9. The rank that could be detected was measured before surgery, approximately 10 min after recovery from anaesthesia, and then 25 min later. After anaesthesia there was a significant increase (P < 0.001) in the rank number by 2, which returned to preoperative values 25 min later. Minute ventilation was reduced by 25% after operation and this may partly explain this change; however, we believe the transient decrease in ability to sense resistance indicates residual effects of anaesthetics and may explain in part the incidence of hypoxia caused by airway obstruction in patients after anaesthesia.


Napping promotes inter-session habituation to emotional stimuli.
The effects of a daytime nap on inter-session habituation to aversive visual stimuli were investigated. Healthy young adult volunteers viewed repeated presentations of highly negative and emotionally neutral (but equally arousing) International Affective Picture System (IAPS) photographs during two afternoon sessions separated by 2.5h. Half of the photographs were shown at both sessions (Repeated Sets) and half differed between sessions (Novel Sets). For each stimulus presentation, evoked skin conductance response (SCR), heart-rate deceleration (HRD) and corrugator supercilii EMG response (EMG), were computed and range corrected using respective maximum session-1 responses. Following each presentation, subjects rated each photograph on dimensions of pleasantness and arousability. During the inter-session interval, Nap subjects had a 120-min polysomnographically monitored sleep opportunity, whereas Wake subjects watched a non-stimulating video. Nap and Wake subjects did not differ in their subjective ratings of photographs. However, for Repeated-Set photographs, Nap subjects demonstrated greater inter-session habituation in SCR and EMG but a trend toward lesser inter-session habituation in HRD. These group differences were absent for Novel-Set photographs. Group differences across all measures were greater for negative stimuli. Occurrence of SWS during the nap was associated with greater inter-session habituation of EMG whereas occurrence of REM was associated with lesser inter-session habituation of SCR to negative stimuli. Sleep may therefore promote emotional adjustment at the level of somatic responses. Physiological but not subjective inter-session habituation to aversive images was enhanced by a daytime nap.


Identification and functional characterization of a novel α-conotoxin (EIIA) from Conus ermineus.
Nicotinic acetylcholine receptors (nAChRs) are one of the most important families in the ligand-gated ion channel superfamily due to their involvement in primordial brain functions and in several neurodegenerative pathologies. The discovery of new ligands which can bind with high affinity and selectivity to nAChR subtypes is of prime interest in order to study these receptors and to potentially discover new drugs for treating various pathologies. Predatory cone snails of the genus Conus hunt their prey using venoms containing a large number of small, highly structured peptides called conotoxins. Conotoxins are classified in different structural families and target a large panel of receptors and ion channels. Interestingly, nAChRs represent the only subgroup for which Conus has developed seven distinct families of conotoxins. Conus venoms have thus received much attention as they could represent a potential source of selective ligands of nAChR subtypes. We describe the mass spectrometric-based approaches which led to the discovery of a novel α-conotoxin targeting muscular nAChR from the venom of Conus ermineus. The presence of several posttranslational modifications complicated the N-terminal sequencing. To discriminate between the different possible sequences, analogs with variable N-terminus were synthesized and fragmented by MS/MS. Understanding the fragmentation pathways in the low m/z range appeared crucial to determine the right sequence. The biological activity of this novel α-conotoxin (α-EIIA) that belongs to the unusual α4/4 subfamily was determined by binding experiments. The results revealed not only its selectivity for the muscular nAChR, but also a clear discrimination between the two binding sites described for this receptor.


Are There Any Clinically Relevant Subgroups of Triple-Negative Breast Cancer in 2018?
The working immunohistochemical definition of triple-negative breast cancer (TNBC) is admittedly reductionist and has only limited usefulness for informing oncologists about therapeutic decisions beyond chemotherapy. Early molecular taxonomies of TNBC based heavily on gene expression profiling, which is not readily available in the clinic today, do not necessarily encompass other molecular targets already incorporated into rationally designed clinical trials. We state that it is possible to delineate five subgroups of TNBC relevant to present-day clinical practice and cover the evidence that lends credence to emerging biomarker-directed treatment strategies for each subgroup.


Understanding racial HIV/STI disparities in black and white men who have sex with men: a multilevel approach.
The reasons for black/white disparities in HIV epidemics among men who have sex with men have puzzled researchers for decades. Understanding reasons for these disparities requires looking beyond individual-level behavioral risk to a more comprehensive framework. From July 2010-December 2012, 803 men (454 black, 349 white) were recruited through venue-based and online sampling; consenting men were provided HIV and STI testing, completed a behavioral survey and a sex partner inventory, and provided place of residence for geocoding. HIV prevalence was higher among black (43%) versus white (13% MSM (prevalence ratio (PR) 3.3, 95% confidence interval (CI): 2.5-4.4). Among HIV-positive men, the median CD4 count was significantly lower for black (490 cells/µL) than white (577 cells/µL) MSM; there was no difference in the HIV RNA viral load by race. Black men were younger, more likely to be bisexual and unemployed, had less educational attainment, and reported fewer male sex partners, fewer unprotected anal sex partners, and less non-injection drug use. Black MSM were significantly more likely than white MSM to have rectal chlamydia and gonorrhea, were more likely to have racially concordant partnerships, more likely to have casual (one-time) partners, and less likely to discuss serostatus with partners. The census tracts where black MSM lived had higher rates of poverty and unemployment, and lower median income. They also had lower proportions of male-male households, lower male to female sex ratios, and lower HIV diagnosis rates. Among black and white MSM in Atlanta, disparities in HIV and STI prevalence by race are comparable to those observed nationally. We identified differences between black and white MSM at the individual, dyadic/sexual network, and community levels. The reasons for black/white disparities in HIV prevalence in Atlanta are complex, and will likely require a multilevel framework to understand comprehensively.


Basis for half-site ligand binding in yeast NAD(+)-specific isocitrate dehydrogenase.
Yeast NAD(+)-specific isocitrate dehydrogenase is an allosterically regulated octameric enzyme composed of four heterodimers of a catalytic IDH2 subunit and a regulatory IDH1 subunit. Despite structural predictions that the enzyme would contain eight isocitrate binding sites, four NAD(+) binding sites, and four AMP binding sites, only half of the sites for each ligand can be measured in binding assays. On the basis of a potential interaction between side chains of Cys-150 residues in IDH2 subunits in each tetramer of the enzyme, ligand binding assays of wild-type (IDH1/IDH2) and IDH1/IDH2(C150S) octameric enzymes were conducted in the presence of dithiothreitol. These assays demonstrated the presence of eight isocitrate and four AMP binding sites for the wild-type enzyme in the presence of dithiothreitol and for the IDH1/IDH2(C150S) enzyme in the absence or presence of this reagent, suggesting that interactions between sulfhydryl side chains of IDH2 Cys-150 residues limit access to these sites. However, only two NAD(+) sites could be measured for either enzyme. A tetrameric form of IDH (an IDH1(G15D)/IDH2 mutant enzyme) demonstrated half-site binding for isocitrate (two sites) in the absence of dithiothreitol and full-site binding (four sites) in the presence of dithiothreitol. Only one NAD(+) site could be measured for the tetramer under both conditions. In the context of the structure of the enzyme, these results suggest that an observed asymmetry between heterotetramers in the holoenzyme contributes to interactions between IDH2 Cys-150 residues and to half-site binding of isocitrate, but that a form of negative cooperativity may limit access to apparently equivalent NAD(+) binding sites.


Aspirations of Latina adolescent suicide attempters.
Parents' aspirations and expectations are communicated to their offspring. Children internalize their parents' aspirations and accept some of the expectations while rejecting others, all part of the developmental process and identity-consolidation. When the aspirations and expectations of youth and parents are incongruent, the outcomes in youths' behavior can be deleterious, such as when adolescents manifest suicidal behaviors. We examined aspirations expressed by 12 Latina adolescent suicide attempters and their parents and compared them to 12 non-suicidal Latinas and parents. Qualitative analyses revealed that incongruence of aspirations between girls and their parents were greater among suicidal teens. Suicidal and non-suicidal Latinas presented contrasting aspirations: the former on gaining independence and the latter on completing their education and pursuing careers. Findings may inform developmental research and ways in which clinicians and policymakers can help Latinas achieve their own and their parents' aspirations.


Characterization of a serine protease homologous to house dust mite group 3 allergens from the scabies mite Sarcoptes scabiei.
The scabies mite, Sarcoptes scabiei var. hominis, infests human skin, causing allergic reactions and facilitating bacterial infection by Streptococcus sp., with serious consequences such as rheumatic fever and rheumatic heart disease. To identify a possible drug target or vaccine candidate protein, we searched for homologues of the group 3 allergen of house dust mites, which we subsequently identified in a cDNA library. The native protein, designated Sar s 3, was shown to be present in the mite gut and excreted in fecal pellets into mite burrows within the upper epidermis. The substrate specificity of proteolytically active recombinant rSar s 3 was elucidated by screening a bacteriophage library. A preference for substrates containing a RS(G/A) sequence at the P1-P2' positions was revealed. A series of peptides synthesized as internally quenched fluorescent substrates validated the phage display data and high performance liquid chromatography/mass spectrometry analysis of the preferred cleaved substrate and confirmed the predicted cleavage site. Searches of the human proteome using sequence data from the phage display allowed the in silico prediction of putative physiological substrates. Among these were numerous epidermal proteins, with filaggrin being a particularly likely candidate substrate. We showed that recombinant rSar s 3 cleaves human filaggrin in vitro and obtained immunohistological evidence that the filaggrin protein is ingested by the mite. This is the first report elucidating the substrate specificity of Sar s 3 and its potential role in scabies mite biology.


Characterization of Listeria monocytogenes isolates from the meat, poultry and seafood industries by automated ribotyping.
A total of 564 Listeria monocytogenes isolates were characterized by automated ribotyping. The samples were taken from equipment, personnel and the environment after cleaning procedures and during food processing, as well as from raw materials and products from six meat, two poultry and five seafood processing plants located in the Faroe Islands, Finland, Iceland, Norway and Sweden. Altogether, 25 different ribotypes (RTs) were generated. Two RTs occurred in the samples from all three food sectors--meat, poultry and seafood. Four RTs occurred in meat and poultry plant samples and other four RTs occurred in meat and seafood plant samples. Five RTs occurred only in meat plant samples, five only in poultry plant samples and five only in seafood plant samples. Eight of the thirteen plants had their own in-house L. monocytogenes ribotype. There was geographical differences between the RTs, but no correlation between RTs and food sectors was detected. The discrimination power of automated ribotyping was satisfactory to trace the contamination sources in the food processing plants clearly indicating the sites at which improved cleaning procedures were necessary. In addition, it was possible to screen a large number of isolates with two instruments located at different institutes and to make a reliable combination of the results.


Antigen bias in T cell cross-priming.
Activated CD8+ T cells detect virally infected cells and tumor cells by recognition of major histocompatibility complex class I-bound peptides derived from degraded, endogenously produced proteins. In contrast, CD8+ T cell activation often occurs through interaction with specialized antigen-presenting cells displaying peptides acquired from an exogenous cellular source, a process termed cross-priming. Here, we observed a marked inefficiency in exogenous presentation of epitopes derived from signal sequences in mouse models. These data indicate that certain virus- and tumor-associated antigens may not be detected by CD8+ T cells because of impaired cross-priming. Such differences in the ability to cross-present antigens should form important considerations in vaccine design.


Potential application of chicken manure biochar towards toxic phenol and 2,4-dinitrophenol in wastewaters.
In this study, chicken manure biochar (CBC) was prepared and applied as adsorbent for the removal of phenolic pollutants including phenol (Ph) and 2,4-Dinitrophenol (DNP) from wastewaters. The feasibility analysis was focused on the adsorption effects of various factors, such as initial concentration, adsorbent dosage and reaction time. The results showed that BC could efficiently remove the Ph and DNP within 90 min of reaction time. Increasing of CBC dosage up to 0.3 g results in the maximum removal efficiency of Ph and DNP and lowers initial concentration which is beneficial for the adsorption of phenolic compounds. The second-order kinetic model and the Langmuir isotherm provided the best correlation with the adsorption data. Based on the Langmuir isotherm, maximum adsorption capacities (qmax) of Ph and DNP were found at 106.2 and 148.1 mg g-1, respectively. The obtained qmax values for CB were higher than those reported in literature on the adsorption of Ph and DNP using different biochar. Analyzing the regeneration characteristics, BC displayed high reusability with less than 20% loss in adsorption capacities of Ph and DNP, even after five repeated cycles. Investigation of the adsorption equilibrium under various conditions suggested several possible interaction mechanisms, including hydrogen bonding, electrostatic interaction and π- π bonding, which were attributed to the binding affinity of the adsorbent-adsorbate interaction. In the field application, the CBC showed an excellent removal efficiencies of Ph and DNP from industrial wastewaters (around 80% phenolic pollutants were removed). These findings support the potential use of CBC as effective adsorbent for treatment of wastewater containing Ph and DNP.


Discovery, optimization and validation of an optimal DNA-binding sequence for the Six1 homeodomain transcription factor.
The Six1 transcription factor is a homeodomain protein involved in controlling gene expression during embryonic development. Six1 establishes gene expression profiles that enable skeletal myogenesis and nephrogenesis, among others. While several homeodomain factors have been extensively characterized with regards to their DNA-binding properties, relatively little is known of the properties of Six1. We have used the genomic binding profile of Six1 during the myogenic differentiation of myoblasts to obtain a better understanding of its preferences for recognizing certain DNA sequences. DNA sequence analyses on our genomic binding dataset, combined with biochemical characterization using binding assays, reveal that Six1 has a much broader DNA-binding sequence spectrum than had been previously determined. Moreover, using a position weight matrix optimization algorithm, we generated a highly sensitive and specific matrix that can be used to predict novel Six1-binding sites with highest accuracy. Furthermore, our results support the idea of a mode of DNA recognition by this factor where Six1 itself is sufficient for sequence discrimination, and where Six1 domains outside of its homeodomain contribute to binding site selection. Together, our results provide new light on the properties of this important transcription factor, and will enable more accurate modeling of Six1 function in bioinformatic studies.


Experimental changes in bodily self-consciousness are tuned to the frequency sensitivity of proprioceptive fibres.
Several lines of evidence suggest an important implication of proprioceptive signals in bodily self-consciousness. By manipulating proprioceptive signals using muscle vibration, here, we investigated whether such effects depend on the vibration frequency by testing three different vibratory stimuli applied at the lower limbs (20, 40 and 80 Hz). We thus explored whether frequency-specific proprioceptive interference that has been reported in postural or motor tasks will also be found for measures of bodily self-consciousness. Self-identification (questionnaires) and visuotactile integration (asking participants to make tactile discriminations) were quantified during synchronous and asynchronous stroking conditions that are known to manipulate bodily self-consciousness. We found that even though muscle vibrations were applied at the same body location in all cases, 20 Hz vibrations did not alter the magnitude of self-identification and visuotactile integration, whereas 40 and 80 Hz vibrations did. These frequency-specific effects extend earlier vibration effects on motor and postural tasks to bodily self-consciousness. We suggest that the observed changes in bodily self-consciousness are due to altered proprioceptive signals from the lower limbs and that these changes depend on the tuning of Ia fibres to muscle vibration.


Staff perception of Lean, care-giving, thriving and exhaustion: a longitudinal study in primary care.
Lean is commonly adopted in healthcare to increase quality of care and efficiency. Few studies of Lean involve staff-related outcomes, and few have a longitudinal design. Thus, the aim was to examine the extent to which changes over time in Lean maturity are associated with changes over time in care-giving, thriving and exhaustion, as perceived by staff, with a particular emphasis on the extent to which job demands and job resources, as perceived by staff, have a moderated mediation effect. A longitudinal study with a correlational design was used. In total, 260 staff at 46 primary care units responded to a web survey in 2015 and 2016. All variables in the study were measured using staff ratings. Ratings of Lean maturity reflect participants' judgements regarding the entire unit; ratings of care-giving, thriving, exhaustion and job demands and resources reflect participants' judgements regarding their own situation. First, over time, increased Lean maturity was associated with increased staff satisfaction with their care-giving and increased thriving, mediated by increased job resources. Second, over time, increased Lean maturity was associated with decreased staff exhaustion, mediated by decreased job demands. No evidence was found showing that job demands and job resources had a moderated mediation effect. The results indicate that primary care staff may benefit from working in organizations characterized by high levels of Lean maturity and that caregiving may also be improved as perceived by staff.


Challenges of accurately measuring and using BMI and other indicators of obesity in children.
BMI is an important indicator of overweight and obesity in childhood and adolescence. When measurements are taken carefully and compared with appropriate growth charts and recommended cutoffs, BMI provides an excellent indicator of overweight and obesity that is sufficient for most clinical, screening, and surveillance purposes. Accurate measurements of height and weight require that adequate attention be given to data collection and management. Choosing appropriate equipment and measurement protocols and providing regular training and standardization of data collectors are critical aspects that apply to all settings in which BMI will be measured and used. Proxy measures for directly measured BMI, such as self-reports or parental reports of height and weight, are much less preferred and should only be used with caution and cognizance of the limitations, biases, and uncertainties attending these measures. There is little evidence that other measures of body fat such as skinfolds, waist circumference, or bioelectrical impedance are sufficiently practicable or provide appreciable added information to be used in the identification of children and adolescents who are overweight or obese. Consequently, for most clinical, school, or community settings these measures are not recommended for routine practice. These alternative measures of fatness remain important for research and perhaps in some specialized screening situations that include a specific focus on risk factors for cardiovascular or diabetic disease.


Validation of the burn intervention score in a National Burn Centre.
The Linköping burn score has been used for two decades to calculate the cost to the hospital of each burned patient. Our aim was to validate the Burn Score in a dedicated Burn Centre by analysing the associations with burn-specific factors: percentage of total body surface area burned (TBSA%), cause of injury, patients referred from other (non-specialist) centres, and survival, to find out which of these factors resulted in higher scores. Our second aim was to analyse the variation in scores of each category of care (surveillance, respiration, circulation, wound care, mobilisation, laboratory tests, infusions, and operation). We made a retrospective analysis of all burned patients admitted during the period 2000-15. Multivariable regression models were used to analyse predictive factors for an increased daily burn score, the cumulative burn score (the sum of the daily burn scores for each patient) and the total burn score (total sum of burn scores for the whole group throughout the study period) in addition to sub-analysis of the different categories of care that make up the burn score. We retrieved 22301 daily recordings for inpatients. Mobilisation and care of the wound accounted for more than half of the total burn score during the study. Increased TBSA% and age over 45 years were associated with increased cumulative (model R2 0.43, p<0.001) and daily (model R2 0.61, p<0.001) burn scores. Patients who died had higher daily burn scores, while the cumulative burn score decreased with shorter duration of hospital stay (p<0.001). To our knowledge this is the first long term analysis and validation of a system for scoring burn interventions in patients with burns that explores its association with the factors important for outcome. Calculations of costs are based on the score, and it provides an indicator of the nurses' workload. It also gives important information about the different dimensions of the care provided from thorough investigation of the scores for each category.


Recruiting Practice-based Research Network (PBRN) physicians to be research participants: lessons learned from the North Texas (NorTex) needs assessment study.
The purpose of this study was to examine strategies for recruiting physician subjects in a practice-based research network continuing education research study, using different recruitment methods at four systems, or health plan arrangements. The North Texas Primary Care Practice-based Research Network Needs Assessment Study consisted of a survey and five self-directed medical record abstractions. Physicians were recruited to be research subjects from four systems, using different recruitment strategies. χ(2) was used to determine differences in physicians consenting and completing the study between systems. Kruskal-Wallis was used to determine differences in time from first contact to consent and number of contacts required before consent between systems. One hundred five of 211 physicians (49.8%) consented to participate, of which 90 (85.7%) completed the survey. There was a significant difference by system in the number of physicians who consented (P = .04) and number of contacts required pre-consent (P < .001) but not in the number of physicians completing the study or time from first contact to consent. Success of recruiting physicians to be research subjects varied between systems using different recruitment methods. Lessons learned include using clinician champions to make initial contact, establishing a relationship with clinic personnel, distinguishing the research team from a pharmaceutical representative, establishing a preferred contact method, and collecting study materials on a set timeline.


The present, past and future of the study of intellectual disability: challenges in developing countries.
There is strong evidence that socio-cultural factors largely determine what is seen as competent behaviour. Within western high income countries, driven by the values of utilitarian individualism, the construct of intellectual disability has been largely determined to meet the needs of urban, industrialised societies. In contrast, competence in non-industrialised societies may be more reflected in collaborative, interpersonal problem solving skills such as those found among Nigerian students labelled as intellectually disabled. However, people who are judged to be incompetent or "obtrusive" in countries deficient in support services, are often neglected and consigned to a life in poorly managed segregated institutions, as is the case in China, Russia and some countries in Eastern Europe. Non western countries that have a long history of a globalised economy, such as Taiwan and Japan also remain committed to segregated institutional provisions for people with an intellectual disability, despite a notional acceptance of inclusionary policies enunciated by the United Nations Declarations and Conventions. In this paper is concluded that it must be recognised that the population of people with an intellectual disability, regardless of how the condition is defined and classified, is quite heterogeneous. Their needs are also varied and not at all dissimilar to those of the general population. As developing countries adopt western style consumer-driven economies, there is an extreme danger that they, too, will follow the same trajectory of exclusion and impose the culture of "otherness" for a group whose contribution to that society will be devalued. Good science is futile unless it benefits all peoples.


Functional optimization of arithmetic processing in perfect performers.
Lesion and imaging studies to date have not clarified which sub-regions of the parietal lobe are specialized for arithmetic processing, and which perform supporting functions. We used functional magnetic resonance imaging to investigate parietal lobe function during arithmetic processing. Functional optimization was examined by analyzing regional differences in brain activation between perfect (100% accuracy) and imperfect performers. Perfect performers had significantly less activation only in the left angular gyrus, a finding that may be associated with skill mastery and long-term practice effects. The present results provide the first direct evidence of localized functional optimization for arithmetic processing in the human brain.


Unexpected sequences and structures of mtDNA required for efficient transcription from the first heavy-strand promoter.
Human mtDNA contains three promoters, suggesting a need for differential expression of the mitochondrial genome. Studies of mitochondrial transcription have used a reductionist approach, perhaps masking differential regulation. Here we evaluate transcription from light-strand (LSP) and heavy-strand (HSP1) promoters using templates that mimic their natural context. These studies reveal sequences upstream, hypervariable in the human population (HVR3), and downstream of the HSP1 transcription start site required for maximal yield. The carboxy-terminal tail of TFAM is essential for activation of HSP1 but not LSP. Images of the template obtained by atomic force microscopy show that TFAM creates loops in a discrete region, the formation of which correlates with activation of HSP1; looping is lost in tail-deleted TFAM. Identification of HVR3 as a transcriptional regulatory element may contribute to between-individual variability in mitochondrial gene expression. The unique requirement of HSP1 for the TFAM tail may enable its regulation by post-translational modifications.


Effect of microcurrent stimulation on delayed-onset muscle soreness: a double-blind comparison.
To examine the efficacy of microcurrent electrical neuromuscular stimulation (MENS) treatment on pain and loss of range of motion (ROM) associated with delayed-onset muscle soreness (DOMS). We assigned subjects to 1 of 2 groups. Group 1 received treatment with microcurrent stimulation (200 muA, 30 Hz, for 10 minutes, then 100 muA, 0.3 Hz, for 10 minutes) 24, 48, and 72 hours after DOMS induction. Group 2 served as a sham group and was treated using a machine altered by the manufacturer so that no current could flow through the electrodes. DOMS was induced in the biceps brachii of the nondominant arm of 18 subjects (3 males, 15 females: age = 20.33 +/- 2.3 years, ht = 170.81 +/- 7.3 cm, wt = 69.61 +/- 13.1 kg). Dominance was defined as the arm used by the subject to throw a ball. Subjective pain and active elbow extension ROM were evaluated before and after treatment each day. Two methods were used to assess pain: constant pressure using a weighted Orthoplast sphere and full elbow extension to the limit of pain tolerance. Subjective pain was measured with a graphic rating scale and active elbow extension ROM using a standard, plastic, double-armed goniometer. Three repeated-measures ANOVAs (between-subjects variable was group, within- subjects variables were day and test) were used to assess ROM and pain scores for the 2 groups. We found no significant difference in the measurement of subjective pain scores or elbow extension ROM when the MENS group was compared with the sham group. Our results indicate that the MENS treatment, within the parameters used for this experiment, was not effective in reducing the pain or loss of ROM associated with delayed-onset muscle soreness.


Building a practically useful theory of goal setting and task motivation. A 35-year odyssey.
The authors summarize 35 years of empirical research on goal-setting theory. They describe the core findings of the theory, the mechanisms by which goals operate, moderators of goal effects, the relation of goals and satisfaction, and the role of goals as mediators of incentives. The external validity and practical significance of goal-setting theory are explained, and new directions in goal-setting research are discussed. The relationships of goal setting to other theories are described as are the theory's limitations.


A combination of genomic approaches reveals the role of FOXO1a in regulating an oxidative stress response pathway.
While many of the phenotypic differences between human and chimpanzee may result from changes in gene regulation, only a handful of functionally important regulatory differences are currently known. As a first step towards identifying transcriptional pathways that have been remodeled in the human lineage, we focused on a transcription factor, FOXO1a, which we had previously found to be up-regulated in the human liver compared to that of three other primate species. We concentrated on this gene because of its known role in the regulation of metabolism and in longevity. Using a combination of expression profiling following siRNA knockdown and chromatin immunoprecipitation in a human liver cell line, we identified eight novel direct transcriptional targets of FOXO1a. This set includes the gene for thioredoxin-interacting protein (TXNIP), the expression of which is directly repressed by FOXO1a. The thioredoxin-interacting protein is known to inhibit the reducing activity of thioredoxin (TRX), thereby hindering the cellular response to oxidative stress and affecting life span. Our results provide an explanation for the repeated observations that differences in the regulation of FOXO transcription factors affect longevity. Moreover, we found that TXNIP is down-regulated in human compared to chimpanzee, consistent with the up-regulation of its direct repressor FOXO1a in humans, and with differences in longevity between the two species.


Quantitative analysis of gingival phenotype in different types of malocclusion in the anterior esthetic zone.
For any esthetic treatment planning, the shape and form of gingiva should be a prime factor of concern. The correct identification of gingival phenotype (GP) ensures a firm foundation for future health and prognosis of the treatment indicated. Hence, the aim of the present study was to evaluate the correlation between the GP in the anterior esthetic zone with different types of maloclussion and severity of crowding. A total of 110 periodontally healthy controls were equally divided into two groups depending on the type of malocclusion. They were further divided according to the levels of dental crowding as mild, moderate, and severe. GP was measured on the anterior esthetic teeth using transgingival probing, and width of the attached gingiva (WAG) was measured using histochemical staining method. In severe crowding group, the GP in 12 and 22 region was found to be thick (P = 0.035) while, in 32 and 42 region was thin (P = 0.042). The WAG shows a significant difference between WAG with 23 in severe crowding group with P = 0.042, whereas there was no significant relationship found between the GP with Angle's classification. Within the limitations of the study, it can be concluded that the teeth in the maxillary and mandibular anterior esthetic region showed the thin phenotype. When the severity of crowding increases, the GP and WAG vary depending on the position of the tooth. There is no association between the Angle's classification and the mean GP of the maxillary and mandibular anterior region teeth.


Investigation of short-term variations in term breast milk composition during repeated breast expression sessions.
Breast milk composition can be affected by several factors, and it can exhibit short-term (weekly) variations. Investigating variations in breast milk composition is important to accurately estimate nutrient requirements of the infant. To investigate short-term changes in breast milk composition between left and right breasts, over a 3-week period within the first 6 months of lactation. The left and right breasts of the mothers of healthy, term infants (n = 23) were simultaneously expressed with an electric breast pump for 15 minutes, on 3 occasions within 3 weeks. Milk samples (5 mL) were collected from the total expression volume of each breast at each session. The macronutrient contents, total solids, and energy content were determined using a mid-infrared human milk analyzer. Mothers (n = 17) measured their 24-hour milk production, and the average 24-hour fat contents were also determined. Over the 3 weekly sessions, no significant changes were found in macronutrient contents. On average, total solids (P = .04) and energy (P = .04) decreased by week 3 of follow-up sessions from 14 to 13 g/100 mL and from 82 to 76 Kcal/100 mL, respectively; however, these changes became insignificant when expression volume was taken into account. The macronutrient concentration was similar for the left and right breasts; however, milk composition varied markedly between mothers. Furthermore, average 24-hour fat content was significantly lower than the mean fat content from a single expression session (P < .01). Our findings highlight that when determining the nutritional adequacy of a mother's milk, assuming an average concentration requires caution. The study findings illustrate the importance of using average 24-hour fat content of milk to obtain representative measures of infant energy intake.


Sepsis-induced depressed contractile function of isolated ventricular myocytes is due to altered calcium transient properties.
Chronic peritoneal sepsis in a rodent model produces myocardial dysfunction characterized by decreased rates of ventricular contraction and relaxation in the isolated heart preparation. However, it remains controversial whether the ventricular contractility is altered during sepsis. In the present study, we determined the effect of chronic peritoneal sepsis on the mechanical properties and intracellular Ca2+ handling of cardiac myocytes isolated from septic rats at 24 or 48 h. Mechanical properties were evaluated by use of an IonOptix MyoCam system. Myocytes were electrically stimulated at 0.5 Hz. The contractile properties analyzed included peak shortening (PS), time-to-peak shortening (TPS), time-to-90% relengthening (TR90), and maximal velocities of shortening and relengthening (+/-dL/dt). Intracellular Ca2+ handling was evaluated with fura-2 fluorescent dye. Myocytes obtained from 24-h postseptic animals exhibited a depressed PS (85% of control), normal TPS, prolonged TR90 (147% of control), and reduced +/-dL/dt (both 79% of control). Myocytes from 48-h postseptic animals also exhibited a reduced peak of intracellular Ca2+ sequestration (55% of control), but resting intracellular Ca2+ and Ca2+-transient decay were comparable with the values seen in myocytes from untreated rats. Myocytes from septic and control animals were equally responsive over a range of stimulation frequencies (0.1-5 Hz). Myocytes from septic animals were unresponsive (5% of control) to increase of extracellular Ca2+ (0.5-3 mM). These results demonstrate that sepsis produces substantial deficits in cardiac myocytes function that can be attributed to altered calcium transient properties.


Binding Modes of Ligands Using Enhanced Sampling (BLUES): Rapid Decorrelation of Ligand Binding Modes via Nonequilibrium Candidate Monte Carlo.
Accurately predicting protein-ligand binding affinities and binding modes is a major goal in computational chemistry, but even the prediction of ligand binding modes in proteins poses major challenges. Here, we focus on solving the binding mode prediction problem for rigid fragments. That is, we focus on computing the dominant placement, conformation, and orientations of a relatively rigid, fragment-like ligand in a receptor, and the populations of the multiple binding modes which may be relevant. This problem is important in its own right, but is even more timely given the recent success of alchemical free energy calculations. Alchemical calculations are increasingly used to predict binding free energies of ligands to receptors. However, the accuracy of these calculations is dependent on proper sampling of the relevant ligand binding modes. Unfortunately, ligand binding modes may often be uncertain, hard to predict, and/or slow to interconvert on simulation time scales, so proper sampling with current techniques can require prohibitively long simulations. We need new methods which dramatically improve sampling of ligand binding modes. Here, we develop and apply a nonequilibrium candidate Monte Carlo (NCMC) method to improve sampling of ligand binding modes. In this technique, the ligand is rotated and subsequently allowed to relax in its new position through alchemical perturbation before accepting or rejecting the rotation and relaxation as a nonequilibrium Monte Carlo move. When applied to a T4 lysozyme model binding system, this NCMC method shows over 2 orders of magnitude improvement in binding mode sampling efficiency compared to a brute force molecular dynamics simulation. This is a first step toward applying this methodology to pharmaceutically relevant binding of fragments and, eventually, drug-like molecules. We are making this approach available via our new Binding modes of ligands using enhanced sampling (BLUES) package which is freely available on GitHub.


A novel approach to acoustic liquid density measurements using a buffer rod based measuring cell.
A new method for measuring the pressure reflection coefficient in a buffer rod configuration is presented, together with experimental results for acoustic measurements of the liquid density, based on the measurement of the liquid's acoustic impedance. The method consists of using 2 buffers enclosing the liquid in a symmetrical arrangement with a transducer fixed to each buffer. One of the transducers is used in a pulse-echo mode while the other transducer operates as a receiver. The echo amplitudes leading to the pressure reflection coefficient as found by this method possess advantages such as reduced attenuation due to a shorter liquid transmission path and reduced interference, as compared with the ABC method. Measurements with distilled water and with special density calibration oil qualities have been performed using both the new method and the ABC method and are shown for the new method to give a density span within +/- 0.15% of the reference values. A comparison of the measured densities based on both a time-domain and a l(2)-norm frequency domain integration signal processing approach is given, along with a recommendation as to how the signal processing should be performed.


Femtosecond laser writing of porous capillaries inside fused silica glass.
We demonstrate that within a restricted optical pulse duration-pulse energy parameter space tightly focused femtosecond laser radiation can be used to fabricate porous capillaries in bulk fused silica glass by simply moving the laser focus through the material. We show that the rate of penetration of liquids into the porous capillaries can be controlled by the laser polarization, which determines their morphology. The fluid propagation is measured using the form birefringence of nanocrack/nanovoid structures produced inside the capillaries. We also demonstrate the nanofiltration capabilities of the capillaries by separating the relatively small molecules of Rhodamine 6G dye from their solvent.


Practice based research networks impacting periodontal care: PEARL Initiative.
In 2005, the National Institute of Dental and Craniofacial Research /National Institutes of Health funded the largest initiative to date to affect change in the delivery of oral care. This commentary provides the background for the first study related to periodontics in a Practice Based Research Network (PBRN). It was conducted in the Practitioners Engaged in Applied Research & Learning (PEARL) Network. The PEARL Network is headquartered at New York University College of Dentistry. The basic tenet of the PBRN initiative is to engage clinicians to participate in clinical studies, where they will be more likely to accept the results and to incorporate the findings into their practices. This process may reduce the translational gap that exists between new findings and the time it takes for them to be incorporated into clinical practice. The cornerstone of the PBRN studies is to conduct comparative effectiveness research studies to disseminate findings to the profession and improve care. This is particularly important because the majority of dentists practice independently. Having practitioners generate clinical data allows them to contribute in the process of knowledge development and incorporate the results in their practice to assist in closing the translational gap. With the advent of electronic health systems on the horizon, dentistry may be brought into the mainstream health care paradigm and the PBRN concept can serve as the skeletal framework for advancing the profession provided there is consensus on the terminology used.


Efficacy and safety of HL301 in the treatment of acute bronchitis and acute exacerbation of chronic bronchitis: a phase 2, randomized, double-blind, placebo-controlled, multicenter study.
The efficacy and safety of Chinese herbs for symptomatic treatment of bronchitis is not well established. We evaluated the efficacy and safety of a combination product of seven herbs (HL301) for the treatment of acute bronchitis (AB) and acute exacerbation of chronic bronchitis (AECB) using a randomized, double-blind, placebo-controlled, multicenter trial design. A total of 160 patients with AB or with AECB were randomized to receive placebo or one of three doses of HL301 (0.6 g/day, 1.2 g/day, or 1.8 g/day) for a total of 7 days. The primary study endpoint was the change in bronchitis severity score (BSS) from the baseline visit (visit 2) to the end of treatment visit (visit 3). Other efficacy variables were percentage BSS systemic sign efficacy after treatment and change in individual BSS parameters after treatment. Changes in BSS from visit 2 to visit 3 in the three treatment groups (4.63 ± 2.24, 4.08 ± 1.63, and 4.15 ± 1.74 in the HL301 0.6 g/day, 1.2 g/day, and 1.8 g/day groups, respectively) were higher than that of the placebo group (2.88 ± 2.57) in the per protocol set (PPS) (P < .05), and it was also valid in the full analysis set (FAS). The number of participants whose symptoms (measured by BSS) improved at least 30% after treatment was higher in all three treatment groups compared to the placebo group in both the FAS and the PPS (P < .05, for all). Three different doses of HL301 (0.6 g/day, 1.2 g/day, and 1.8 g/day) were effective in decreasing the BSS index compared to placebo. HL301 may be effective for symptomatic treatment of both AB and AECB. Essential components of HL301 have not been delineated in the study and patients with AB and AECB were indiscriminately enrolled in the present study. Respective evaluation of the efficacy of HL301 for AB and AECB will be necessary in the future.


Bacillus wiedmannii biovar thuringiensis: A Specialized Mosquitocidal Pathogen with Plasmids from Diverse Origins.
Bacillus cereus sensu lato also known as B. cereus group is composed of an ecologically diverse bacterial group with an increasing number of related species, some of which are medically or agriculturally important. Numerous efforts have been undertaken to allow presumptive differentiation of B. cereus group species from one another. FCC41 is a Bacillus sp. strain toxic against mosquito species like Aedes aegypti, Aedes (Ochlerotatus) albifasciatus, Culex pipiens, Culex quinquefasciatus, and Culex apicinus, some of them responsible for the transmission of vector-borne diseases. Here, we report the complete genome sequence of FCC41 strain, which consists of one circular chromosome and eight circular plasmids ranging in size from 8 to 490 kb. This strain harbors six crystal protein genes, including cry24Ca, two cry4-like and two cry52-like, a cry41-like parasporin gene and multiple virulence factors. The phylogenetic analysis of the whole-genome sequence of this strain with molecular approaches places this strain into the Bacillus wiedmannii cluster. However, according with phenotypical characteristics such as the mosquitocidal activity due to the presence of Cry proteins found in the parasporal body and cry genes encoded in plasmids of different sizes, indicate that this strain could be renamed as B. wiedmannii biovar thuringiensis strain FCC41.


Exploring the clinical utility of blood ketone levels in the emergency department assessment of paediatric patients.
Ketonuria (on standard urine testing) is a frequent finding in children presenting to emergency departments. With the advent of hand-held ketone meters, blood ketone levels can now be rapidly quantified. Point of care testing (POCT) of blood ketone levels could provide clinically useful information on severity of illness in children and risk of hospital admission. A prospective study using POCT of blood ketone levels in a convenience sample of children <13 years old, with a typical case mix of medical problems. 186 children were studied. The range of ketone levels varied widely among this study population depending on the presenting complaint. Higher levels were noted in those presenting with anorexia or vomiting and fever. The median ketone level of the total study population was 0.2 (range 0-6.0, interquartile range 0.1-0.9) mmol/l. Ketone levels correlated poorly with discharge destination and duration of admission. However, receiver-operator characteristics for ketones as a predictor of admission were comparable to Pediatric Risk of Admission scores (area under the curve 0.64 and 0.72, respectively) and may represent an independent risk factor for admission. A ketone level >1.2 mmol/l has a positive predictive value of 66.7% for admission. Ketone levels correlated well with decreased oral intake (R2 = 0.25; p<0.001). A strong association was found between ketone levels, decreased oral intake and fever. Although ketone levels do not correlate well with more traditional markers of illness severity, they can help to predict the requirement for admission to hospital when interpreted in the context of the presenting illness. They may have applications in both the emergency department and primary care settings. Further prospective testing is required to validate these findings.


Seasonality and temporal correlation between community antibiotic use and resistance in the United States.
Therapeutic antibiotic use in humans is a significant driver of antibiotic resistance. The seasonal effect of antibiotic use on antibiotic resistance has been poorly quantified because of lack of large-scale, spatially disaggregated time-series data on antibiotic use and resistance. We used time-series analysis (Box-Jenkins) on US antibiotic usage from IMS Health and on antibiotic resistance from The Surveillance Network from 1999-2007 to estimate the effect of aminopenicillin, fluoroquinolone, trimethoprim/sulfamethoxazole, and tetracycline usage on resistance of Escherichia coli to drugs within these classes. We also quantified the effect of fluoroquinolone and macrolide/lincosamide usage on resistance of methicillin-resistant Staphylococcus aureus (MRSA) to ciprofloxacin and clindamycin (which has a similar mode of action to macrolides), respectively. Prevalence of resistant Escherichia coli was significantly correlated with lagged (by 1 month) antibiotic prescriptions for aminopenicillins (0.22, P = .03) and fluoroquinolones (0.24, P = .02), which are highly prescribed, but was uncorrelated to antibiotic classes with lower prescription levels. Fluoroquinolone prescriptions were also significantly correlated with a 1-month lag with the prevalence of ciprofloxacin-resistant MRSA (0.23, P = .03). Large-scale usage of antibiotics can generate seasonal patterns of resistance that fluctuate on a short time scale with changes in antibiotic retail sales, suggesting that use of antibiotics in the winter could have a significant effect on resistance. In addition, the strong correlation between community use of antibiotics and resistance isolated in the hospital indicates that restrictions imposed at the hospital level are unlikely to be effective unless coordinated with campaigns to reduce unnecessary antibiotic use at the community level.


Spatial variations in the consumption of illicit stimulant drugs across Australia: A nationwide application of wastewater-based epidemiology.
Obtaining representative information on illicit drug use and patterns across a country remains difficult using surveys because of low response rates and response biases. A range of studies have used wastewater-based epidemiology (WBE) as a complementary approach to monitor community-wide illicit drug use. In Australia, no large-scale WBE studies have been conducted to date to reveal illicit drug use profiles in a national context. In this study, we performed the first Australia-wide WBE monitoring to examine spatial patterns in the use of three illicit stimulants (cocaine, as its human metabolite benzoylecgonine; methamphetamine; and 3,4-methylendioxymethamphetamine (MDMA)). A total of 112 daily composite wastewater samples were collected from 14 wastewater treatment plants across four states and two territories. These covered approximately 40% of the Australian population. We identified and quantified illicit drug residues using liquid chromatography coupled with tandem mass spectrometry. There were distinctive spatial patterns of illicit stimulant use in Australia. Multivariate analyses showed that consumption of cocaine and MDMA was higher in the large cities than in rural areas. Also, cocaine consumption differed significantly between different jurisdictions. Methamphetamine consumption was more similar between urban and rural locations. Only a few cities had elevated levels of use. Extrapolation of the WBE estimates suggested that the annual consumption was 3tonnes for cocaine and 9tonnes combined for methamphetamine and MDMA, which outweighed the annual seizure amount by 25 times and 45 times, respectively. These ratios imply the difficulty of detecting the trafficking of these stimulants in Australia, possibly more so for methamphetamine than cocaine. The obtained spatial pattern of use was compared with that in the most recent national household survey. Together both WBE and survey methods provide a more comprehensive evaluation of drug use that can assist governments in developing policies to reduce drug use and harm in the communities.


Warm-up, stretching and massage diminish harmful effects of eccentric exercise.
The effect of a combination of a warm-up, stretching exercises and massage on subjective scores for delayed onset muscle soreness (DOMS) and objective functional and biochemical measures was studied. Fifty people, randomly divided in a treatment and a control group, performed eccentric exercise with the forearm flexors for 30 min. The treatment group additionally performed a warm-up and underwent a stretching protocol before the eccentric exercise and massage afterwards. Functional and biochemical measures were obtained before, and 1, 24, 48, 72 and 96h after exercise. The median values at the five post-exercise time points differed significantly for DOMS measured when the arm was extended (p = 0.043). Significant main effects for treatment were found on the maximal force (p = 0.026), the flexion angle of the elbow (p = 0.014) and the creatine kinase activity in blood (p = 0.006). No time-by-treatment interactions were found. DOMS on pressure, extension angle and myoglobin concentration in blood did not differ between the groups. This combination of a warm-up, stretching and massage reduces some negative effects of eccentric exercise, but the results are inconsistent, since some parameters were significantly affected by the treatment whereas others were not, despite the expected efficacy of a combination of treatments. The objective measures did not yield more unequivocal results than the subjective DOMS scores.


Successful treatment of low-grade endometrial cancer in premenopausal women with an aromatase inhibitor after failure with oral or intrauterine progesterone.
Young women with endometrial intraepithelial hyperplasia or low-grade endometrial carcinoma are potential candidates for conservative fertility sparing therapy utilizing progesterone rather than hysterectomy. High-dose progesterone treatment is associated with 55-80% initial response but high relapse rates. Using aromatase inhibitors in conjunction with high-dose progesterone has largely been unstudied. Three obese premenopausal women with endometrial cancer failed to respond to oral or intrauterine progesterone as first line therapy. Due to their desire to continue to pursue fertility sparing treatment options, an aromatase inhibitor was added to their treatment regimen. This resulted in resolution of their malignancy in each case. In obese premenopausal women, the mechanism of malignant transformation in endometrial carcinoma is considered to be an association with relatively high levels of serum estrogen from peripheral conversion of androgens to estrone in adipose tissue with a deficiency in progesterone exposure due to chronic anovulation. Using aromatase inhibitors seems reasonable as an adjunct to progesterone given the high likelihood that this population has a significant proportion of their estrogen production coming from peripheral conversion in adipose tissue. This case series is unique in that each woman initially failed to respond to progesterone but had resolution when an aromatase inhibitor was added to their treatment regimen. This would suggest that obese women with low grade malignancy or hyperplasia who have no radiographic evidence of deep myometrial invasion, ovarian or retroperitoneal metastases and who wish to retain their fertility may be treated with intrauterine progesterone and an aromatase inhibitor.


Autocatalytic replication and homochirality in biopolymers: is homochirality a requirement of life or a result of it?
A key step in the origin of life is the establishment of autocatalytic cycles controlled by biopolymer catalysts. These catalysts (either ribozymes or proteins) are composed of homochiral monomers. Homochirality in living systems is maintained because biopolymers are asymmetric in their catalysis and synthesize molecules of their own handedness. Asymmetric autocatalysis is also possible with small molecules, as demonstrated by the Soai reaction, but it is rare. As far as we know, single nucleotides and amino acids are not autocatalytic. The observation that organic molecules in meteorites can have an enantiomeric excess of a few percent suggests that the prebiotic mixture may have had a partial chiral bias that was caused by external physical influences. Here, we consider the way that such a partial prebiotic bias would influence the origin of ribozymes in an RNA world scenario. We have previously shown how a transition to a living state can occur in a model for RNA polymerization. Here, we add chirality to the problem by considering simultaneous synthesis and polymerization of left- and right-handed monomers. The two chemical synthesis rates may be equal or unequal, due to physical or chemical effects prior to the origin of life. We determine the stationary states of this reaction system. The nonliving state is racemic, or slightly biased. There are two living states that are almost completely homochiral, whether or not the nonliving state is biased. It is a feature of our model that, for some regions of parameter space, living and nonliving states are both found to be stable under the same conditions. The origin of life therefore involves a stochastic transition between the nonliving and living states. Our model extends previous theories by treating the origin of life and the origin of chirality as aspects of the same model.


Evaluating an audit and feedback intervention for reducing antibiotic prescribing behaviour in general dental practice (the RAPiD trial): a partial factorial cluster randomised trial protocol.
Antibiotic prescribing in dentistry accounts for 9% of total antibiotic prescriptions in Scottish primary care. The Scottish Dental Clinical Effectiveness Programme (SDCEP) published guidance in April 2008 (2nd edition, August 2011) for Drug Prescribing in Dentistry, which aims to assist dentists to make evidence-based antibiotic prescribing decisions. However, wide variation in prescribing persists and the overall use of antibiotics is increasing. RAPiD is a 12-month partial factorial cluster randomised trial conducted in NHS General Dental Practices across Scotland. Its aim is to compare the effectiveness of individualised audit and feedback (A&F) strategies for the translation into practice of SDCEP recommendations on antibiotic prescribing. The trial uses routinely collected electronic healthcare data in five aspects of its design in order to: identify the study population; apply eligibility criteria; carry out stratified randomisation; generate the trial intervention; analyse trial outcomes. Eligibility was determined on contract status and a minimum level of recent NHS treatment provision. All eligible dental practices in Scotland were simultaneously randomised at baseline either to current audit practice or to an intervention group. Randomisation was stratified by single-handed/multi-handed practices. General dental practitioners (GDPs) working at intervention practices will receive individualised graphical representations of their antibiotic prescribing rate from the previous 14 months at baseline and an update at six months. GDPs could not be blinded to their practice allocation. Intervention practices were further randomised using a factorial design to receive feedback with or without: a health board comparator; a supplementary text-based intervention; additional feedback at nine months. The primary outcome is the total antibiotic prescribing rate per 100 courses of treatment over the year following delivery of the baseline intervention. A concurrent qualitative process evaluation will apply theory-based approaches using the Consolidated Framework for Implementation Research to explore the acceptability of the interventions and the Theoretical Domains Framework to identify barriers and enablers to evidence-based antibiotic prescribing behaviour by GDPs. RAPiD will provide a robust evaluation of A&F in dentistry in Scotland. It also demonstrates that linked administrative datasets have the potential to be used efficiently and effectively across all stages of an randomised controlled trial. Current Controlled Trials ISRCTN49204710.


On average, physicians spend nearly 11 percent of their 40-year careers with an open, unresolved malpractice claim.
The US malpractice system is widely regarded as inefficient, in part because of the time required to resolve malpractice cases. Analyzing data from 40,916 physicians covered by a nationwide insurer, we found that the average physician spends 50.7 months-or almost 11 percent-of an assumed forty-year career with an unresolved, open malpractice claim. Although damages are a factor in how doctors perceive medical malpractice, even more distressing for the doctor and the patient may be the amount of time these claims take to be adjudicated. We conclude that this fact makes it important to assess malpractice reforms by how well they are able to reduce the time of malpractice litigation without undermining the needs of the affected patient.


Evaluation of p16/Ki-67 dual-stained cytology as triage test for high-risk human papillomavirus-positive women.
The aim of this study was to evaluate the clinical utility of p16/Ki-67 dual staining, for the identification of CIN in high-risk HPV-positive women from a non-responder screening cohort. P16/Ki-67 dual staining, Pap cytology, and HPV16/18 genotyping were performed on physician-taken liquid-based samples from 495 women who tested high-risk HPV positive on self-sampled material (PROHTECT-3B study). Different triage strategies involving p16/Ki-67 dual staining were evaluated for sensitivity, specificity, and predictive value for ≥CIN2 and ≥CIN3, and compared to Pap cytology with a threshold of atypical cells of undetermined significance. Centrally revised histology or an adjusted endpoint with combined high-risk HPV negative and cytology negative follow-up at 6 months was used as gold standard. Pap cytology (threshold atypical cells of undetermined significance) triage of high-risk HPV-positive samples showed a sensitivity of 93% (95% confidence interval: 85-98) with a specificity of 49% (95% confidence interval: 41-56) for ≥CIN3. Three triage strategies with p16/Ki-67 showed a significantly increased specificity with similar sensitivity. P16/Ki-67 triage of all high-risk HPV-positive samples had a sensitivity of 92% (95% confidence interval: 84-97) and a specificity of 61% (95% confidence interval: 54-69) for ≥CIN3. Applying p16/Ki-67 triage to only high-risk HPV-positive women with low-grade Pap cytology showed a similar sensitivity of 92% (95% confidence interval: 84-97), with a specificity for ≥CIN3 of 64% (95% confidence interval: 56-71). For high-risk HPV-positive women with low-grade and normal Pap cytology, triage with p16/Ki-67 showed a sensitivity of 96% (95% confidence interval: 89-99), and a specificity of 58% (95% confidence interval: 50-65). HPV16/18 genotyping combined with Pap cytology showed a sensitivity and specificity for ≥CIN3 similar to Pap cytology with an atypical cells of undetermined significance threshold. Because the quality of Pap cytology worldwide varies, and differences in sensitivity and specificity are limited between the three selected strategies, p16/Ki-67 triage of all high-risk HPV-positive samples would be the most reliable strategy in triage of high-risk HPV-positive women with an increased specificity and similar sensitivity compared with Pap cytology triage.