Genetic variation in CCR2 and CXCL12 genes impacts on CD4 restoration in patients initiating cART with advanced immunesupression.
We investigated the association of genetic polymorphisms in chemokine and chemokine receptor genes with poor immunological recovery in HIV patients starting combined antiretroviral therapy (cART) with low CD4 T-cell counts. A case-control study was conducted in 412 HIV-infected patients starting cART with CD4 T-cell count <200 cells/μL and successful viral control for two years. CD4 count increase below 200 cells/μL after two years on cART was used to define INR (immunological non-responder) patients. Polymorphisms in CXCL12, CCL5 and CCR2 genes were genotyped using sequenom's MassARRAY platform. Thirty two percent (134/412) of patients were classified as INR. After adjusting by age, route of HIV infection, length of infection before cART and viral hepatitis coinfection, CCR2 rs1799864-AG genotype was significantly associated with INR status (OR [95% CI]: 1.80 [1.04-3.11]; p = 0.04), and CXCL12 rs1801157-TT genotype showed a trend (OR [95% CI]: 2.47 [0.96-6.35]; p = 0.06). CCR2 rs1799864-AG or CXCL12 rs1801157-TT genotypes influence on the probability of poor CD4 recovery in the population of HIV patients starting cART with low CD4 counts. Genotyping of these polymorphisms could be used to estimate the risk of poor CD4 restoration, mainly in patients who are diagnosed late in the course of infection.


Clinicians' experiences of anxiety in patients with dementia.
Since anxiety in patients with dementia is a complex, understudied phenomenon, this paper presents clinicians' experiences of anxiety in this population. Semi-structured interviews were conducted with seven clinicians experienced with dementia in elderly patients (65 years and above), and then evaluated via qualitative content analysis. Analysis revealed three main categories: A reaction to loss and worries, symptoms of anxiety and depression interfere with each other, and anxiety in dementia-a multidisciplinary task. Anxiety in this population is perhaps best understood as a reaction to loss and worries, and existential in nature by the participants. Care interventions can reduce or prevent anxiety symptoms in this population. However, when anxiety co-exists with depression it might be difficult to attenuate these symptoms through care measures alone. To better identify and treat the condition, valid dementia-specific anxiety-screening instruments are necessary.


Location of the active site of the bean alpha-amylase inhibitor and involvement of a Trp, Arg, Tyr triad.
Seeds of the common bean contain three homologous proteins: phytohaemagglutinin E, phytohaemagglutinin L and the lectin-like protein alpha-amylase inhibitor (alpha AI). Whereas the active site of lectins has been studied in great detail, there is no information on the active site of the related protein alpha AI, which exerts its biological activity by making a 1:1 complex with alpha-amylase. alpha-amylase inhibitor is synthesized as a 30 kDa precursor glycoprotein that needs to be processed at Asn77 to form an active molecule. Comparison of the amino acid sequence of the bean alpha AI with that of the bacterial amylase inhibitor, tendamistat, suggested that a region around Trp188 might be involved in the inhibitory site. When a three-dimensional model of the bean alpha AI was constructed based on its homology to the legume lectins, this Trp region was alongside Asn77. To test this site hypothesis, mutants of alpha AI were created by site-directed mutagenesis of the cDNA and expressed in transgenic tobacco. The mutant proteins R74N and WSY188-190GNV, as well as the double mutant, were inactive as inhibitors. These findings suggest that the active site of alpha AI consists of W188, R74 and Y190, in analogy to the Trp-Arg-Tyr motif of tendamistat, and that the processing of the polypeptide at Asn77 may be necessary to bring these residues in close proximity.


Evaluating dynamic effects of copy number alterations on gene expression using a single transcription model.
DNA copy number alterations (CNAs) are known to be related to genetic diseases, including cancer. The unlimited transcription (UT) model, in which transcription occurs permissively with a simple activation probability, has been proposed to investigate long-term effects of CNAs on gene expression values. Queueing theory was applied, and the copy-number-gene-expression relationship has been shown to be generally nonlinear in the UT model. However, the dynamic effects of CNAs on transcription and the underlying disorders related to diseases remain greatly unknown. Since most genes in a single cell are permissively transcribed in short periods of time interspersed by long periods of limited transcription, an alternative model for transcription in the restrictive state is needed for unraveling the effects of CNAs on gene expression levels with time. To address these issues, herein a single transcription (ST) model is proposed, in which bound TFs are assumed to be unloaded immediately after stimulating a transcription. Using the Laplace-Stieltjes transform and numerical analysis, the relationship between DNA copy number and gene expression level is evaluated. Dynamic modeling reveals that CNAs would potentially alter, or even reverse, the burst-like gene expression modifications while shifting from the ST model to the UT model. Moreover, functional disorders in transcriptional oscillation due to CNAs are shown via simulation. This paper demonstrates how mathematical theories could be helpful to interpret statistical findings from real data and achieve a better understanding of cancer biology.


Preventive effects of garlic (Allium sativum) on oxidative stress and histopathology of cardiac tissue in streptozotocin-induced diabetic rats.
Since some complications of diabetes mellitus may be caused or exacerbated by an oxidative stress, the protective effects of garlic (Allium sativum) were investigated in the blood and heart of streptozotocin-induced diabetic rats. Twenty-eight male Wistar rats were randomly divided into four groups: control, garlic, diabetic, and diabetic+garlic. Diabetes was induced by intraperitoneal (i.p.) injection of streptozotocin (50 mg/kg) in male rats. Rats were fed with raw fresh garlic homogenate (250 mg/kg) six days a week by gavage for a period of 6 weeks. At the end of the 6th week blood samples and heart tissues were collected and used for determination of glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) and histological evaluation. Induction of diabetes increased MDA levels in blood and homogenates of heart. In diabetic rats treated with garlic, MDA levels decreased in blood and heart homogenates. Treatment of diabetic rats with garlic increased SOD, GPX and CAT in blood and heart homogenates. Histopathological finding of the myocardial tissue confirmed a protective role for garlic in diabetic rats. Thus, the present study reveals that garlic may effectively modulate antioxidants status in the blood and heart of streptozotocin induced-diabetic rats.


Demonstration of rare protein in the outer membrane of Treponema pallidum subsp. pallidum by freeze-fracture analysis.
The surface of Treponema pallidum subsp. pallidum (T. pallidum), the etiologic agent of syphilis, appears antigenically inert and lacks detectable protein, as judged by immunocytochemical and biochemical techniques commonly used to identify the outer membrane (OM) constituents of gram-negative bacteria. We examined T. pallidum by freeze-fracture electron microscopy to visualize the architecture of its OM. Treponema phagedenis biotype Reiter (T. phagedenis Reiter), a nonpathogenic host-associated treponeme, and Spirochaeta aurantia, a free-living spirochete, were studied similarly. Few intramembranous particles interrupted the smooth convex and concave fracture faces of the OM of T. pallidum, demonstrating that the OM of this organism is an unusual, nearly naked lipid bilayer. In contrast, the concave fracture face of the OM of S. aurantia was densely covered with particles, indicating the presence of abundant integral membrane proteins, a feature shared by typical gram-negative organisms. The concentration of particles in the OM concave fracture face of T. phagedenis Reiter was intermediate between those of T. pallidum and S. aurantia. Similar to typical gram-negative bacteria, the OM convex fracture faces of the three spirochetes contained relatively few particles. The unique molecular architecture of the OM of T. pallidum can explain the puzzling in vitro properties of the surface of the organism and may reflect a specific adaptation by which treponemes evade the host immune response.


Communication in Plants: Comparison of Multiple Action Potential and Mechanosensitive Signals With Experiments.
Both action potentials and mechanosensitive signalling are an important communication mechanisms in plants. Considering an information-theoretic framework, this paper explores the effective range of multiple action potentials for a long chain of cells (i.e., up to 100) in different configurations, and introduces the study of multiple mechanosensitive activation signals (generated due to a mechanical stimulus) in plants. For both these signals, we find that the mutual information per cell and information propagation speed tends to increase up to a certain number of receiver cells. However, as the number of cells increase beyond 10 to 12, the mutual information per cell starts to decrease. To validate our model and results, we include an experimental verification of the theoretical model, using a PhytlSigns biosignal amplifier, allowing us to measure the magnitude of the voltage associated with the multiple AP's and mechanosensitive activation signals induced by different stimulus in plants. Experimental data is used to calculate the mutual information and information propagation speed, which is compared with corresponding numerical results. Since these signals are used for a variety of important tasks within the plant, understanding them may lead to new bioengineering methods for plants.


Organic cultivation of Triticum turgidum subsp. durum is reflected in the flour-sourdough fermentation-bread axis.
Triticum turgidum subsp. durum was grown according to four farming systems: conventional (CONV), organic with cow manure (OMAN) or green manure (OLEG), and without inputs (NOINPUT). Some chemical and technological characteristics differed between CONV and organic flours. As shown by two-dimensional electrophoresis (2-DE) analysis, OMAN and OLEG flours showed the highest number of gliadins, and OMAN flour also had the highest number of high-molecular-mass glutenins. Type I sourdoughs were prepared at the laboratory level through a back-slopping procedure, and the bacterial ecology during sourdough preparation was described by 16S rRNA gene pyrosequencing. Before fermentation, the dough made with CONV flour showed the highest bacterial diversity. Flours were variously contaminated by genera belonging to the Proteobacteria, Firmicutes, and Actinobacteria. Mature sourdoughs were completely and stably dominated by lactic acid bacteria. The diversity of Firmicutes was the highest for mature sourdoughs made with organic and, especially, NOINPUT flours. Beta diversity analysis based on the weighted UniFrac distance showed differences between doughs and sourdoughs. Those made with CONV flour were separated from the other with organic flours. Lactic acid bacterium microbiota structure was qualitatively confirmed through the culturing method. As shown by PCR-denaturing gradient gel electrophoresis (DGGE) analysis, yeasts belonging to the genera Saccharomyces, Candida, Kazachstania, and Rhodotorula occurred in all sourdoughs. Levels of bound phenolic acids and phytase and antioxidant activities differed depending on the farming system. Mature sourdoughs were used for bread making. Technological characteristics were superior in the breads made with organic sourdoughs. The farming system is another determinant affecting the sourdough microbiota. The organic cultivation of durum wheat was reflected along the flour-sourdough fermentation-bread axis.


Lack of association between aspirin responsiveness and seven candidate gene haplotypes in patients with symptomatic vascular disease.
We studied the effect of prophylactic aspirin (ASA) ingestion on platelet function in 463 patients with stroke, transient ischemic attack (TIA) or acute coronary disease (ACD), using the Platelet Function Analyzer-100 (PFA-100). We correlated ASA responsiveness with haplotypes of seven candidate genes, selected for their documented role in platelet function, namely, the genes for integrins alpha2beta1and alphaIIbbeta3 (ITGA2, ITGA2B, and ITGB3), platelet glycoproteins Ibalpha and VI (GPIBA and GP6), the purinergic receptor P2Y1 (P2RY1), and prostaglandin H synthase 1 (PTGS1 = COX1). Non-responsiveness to ASA was defined as the failure of prior ASA ingestion to prolong the PFA-100 closure time (CT) when blood was perfused through cartridges coated with collagen plus epinephrine (CEPI-CT). ASA non-responsiveness was observed in 114 of 463 patients (24.6 %), but was not associated with haplotypes of any of the seven candidate genes. There was also no association between any haplotypes and the CT when blood was perfused through cartridges coated with collagen plus ADP (CADP-CT). The ASA non-responsive cohort had significantly increased whole blood platelet counts (p = 0.03) and plasma von Willebrand Factor antigen levels (p < 0.001), which likely contributes to resistance to the inhibitory effects of ASA in the PFA-100.


Association of allostatic load with brain structure and cognitive ability in later life.
Allostatic load (AL) has been proposed as a general framework for understanding the cumulative effects of life stress on individuals. Despite growing interest in AL, limited research has been conducted on aging samples. We consider the association of AL (operationalized by a range of inflammatory, cardiovascular, and metabolic measures) with a range of brain volume measurements and cognitive ability in a large cohort sample of older adults (n = 658, mean age = 72.5 years, standard deviation = 0.7) using structural equation modeling. AL was significantly inversely associated with total brain volume (range of standardized β = -0.16 to -0.20) and white-matter volume (-0.35 to -0.36) and positively with hippocampal volume (0.10-0.15) but not gray-matter volume (0.04). AL was also significantly inversely associated with general cognitive ability (range β = -0.13 to -0.20), processing speed (-0.20 to -0.22), and knowledge (-0.18 to -0.20) but not memory or nonverbal reasoning. The associations of AL with cognitive abilities were not mediated by these brain volume measures. AL did not predict cognitive change from age 11 to approximately age 73. The findings suggest a link between AL and later life brain health and cognitive functioning.


Isolated intraocular relapses of primary cerebral lymphomas: An LOC network study.
Most relapses of primary central nervous system lymphoma (PCNSL) occur in the brain and are associated with a poor prognosis. Isolated intraocular relapses (IIORs) are rare and poorly described. We retrospectively selected from the French Lymphome Oculo-Cérébral database PCNSL patients who initially presented with cerebral localization and who experienced IIOR during the course of the disease. Of the 1472 patients included in the database, 55 patients presented an IIOR. Their median age was 68 years, and median Karnofsky Performance Status 80. IL-10 levels in the aqueous humor and/or in the vitreous were increased in 42/46 patients. 45/55 patients received systemic chemotherapy, and 11/55 received high-dose chemotherapy with autologous stem cell transplantation (HCT-ASCT) as consolidation treatment. After a median follow-up of 69 months, 42/55 patients had relapsed, including 90% of the patients who did not receive HCT-ASCT at IIOR and 40% of the patients who received HCT-ASCT at IIOR (p < 0.001). The first relapse after the initial IIOR was exclusively in the eye in 23/42 patients, and 29/42 patients had a subsequent brain relapse during the course of the disease. The median progression-free survival, brain-free survival and overall survival from IIOR were 12.2, 48.6 and 57.1 months, respectively. Isolated intraocular relapse is not exceptional in the course of PCNSL and deserves systematic ophthalmological follow-up. Its prognosis is much better than the prognosis of brain relapse, with an evolution close to that of primary vitreoretinal lymphoma. With the exception of patients who received HCT-ASCT at IIOR, almost all patients subsequently relapsed, often with other IIORs.


Bivariate Analysis of Age-Related Macular Degeneration Progression Using Genetic Risk Scores.
Age-related macular degeneration (AMD) is a leading cause of blindness in the developed world. While many AMD susceptibility variants have been identified, their influence on AMD progression has not been elucidated. Using data from two large clinical trials, Age-Related Eye Disease Study (AREDS) and AREDS2, we evaluated the effects of 34 known risk variants on disease progression. In doing so, we calculated the eye-level time-to-late AMD and modeled them using a bivariate survival analysis approach, appropriately accounting for between-eye correlation. We then derived a genetic risk score (GRS) based on these 34 risk variants, and analyzed its effect on AMD progression. Finally, we used the AREDS data to fit prediction models of progression based on demographic and environmental factors, eye-level AMD severity scores and the GRS and tested the models using the AREDS2 cohort. We observed that GRS was significantly associated with AMD progression in both cohorts, with a stronger effect in AREDS than in AREDS2 (AREDS: hazard ratio (HR) = 1.34, P = 1.6 × 10-22; AREDS2: HR = 1.11, P = 2.1 × 10-4). For prediction of AMD progression, addition of GRS to the demographic/environmental risk factors considerably improved the prediction performance. However, when the baseline eye-level severity scores were included as the predictors, any other risk factors including the GRS only provided small additional predictive power. Our model for predicting the disease progression risk demonstrated satisfactory performance in both cohorts, and we recommend its use with baseline AMD severity scores plus baseline age, education level, and smoking status, either with or without GRS.


CT evaluation of musculoskeletal trauma: initial experience with cinematic rendering.
Multi-detector computed tomography (MDCT) scanners that can quickly acquire volumetric datasets composed of isotropic voxels laid the groundwork for the widespread clinical implementation of 3D MDCT reconstructions, with maximum intensity projection (MIP) and volumetric rendering (VR) becoming important parts of the imaging evaluation of patients with a wide variety of pathologic conditions. Recently, a new 3D reconstruction technique known as cinematic rendering (CR) has become available and is now U.S. FDA approved. CR bears fundamental similarities to VR, but utilizes a more complex lighting model to bring about photorealistic reconstructions. While a tremendous amount of work remains to be done in order to understand the advantages and disadvantages of CR in comparison to traditional 3D reconstruction methods, the images themselves are strikingly detailed and can be interactively manipulated to highlight a variety of different tissue types and anatomic structures. In the following pictorial essay, we provide a number of clinical examples of the use of CR in musculoskeletal imaging, including the evaluation of complex fractures, the delineation of the relationship of fractures to adjacent vasculature and overlying soft tissues, and the visualization of vascular and soft tissue injuries.


Evaluation of a novel kallikrein inhibitor on hemostatic activation in vitro.
DX-88 is a potent kallikrein inhibitor that is being studied for the treatment of hereditary angioedema (HAE) and represents a potential alternative to aprotinin in cardiac surgical patients. The current study was designed to evaluate in vitro effects of DX-88 on coagulation in comparison with aprotinin. Blood samples were obtained from consented 12 healthy volunteers. DX-88 or aprotinin was added to blood at 200 and 800 kallikrein inhibitory units (KIU) per milliliter for aprotinin, and at 1.1, 2.2, or 8.8 microg/ml for DX-88. Thromboelastography (TEG) was performed using celite, kaolin, or tissue factor (TF) activation. Kaolin-based activated clotting times (ACTs) were measured at different heparin levels. The whole blood prothrombin time (PT)/PTT values were also measured. The endogenous thrombin generation was assessed with a fluorogenic assay using platelet-poor plasma (PPP). With celite and kaolin activation of TEG, the reaction time was prolonged with DX-88 and aprotinin. With tissue factor activation, TEG parameters were not affected. DX-88 caused dose-dependent kaolin-ACT prolongation that was augmented by increasing doses of heparin. DX-88 or aprotinin had no significant effects on the PT values, but PTT values were dose-dependently prolonged. Both agents delayed the onset of thrombin generation when PTT reagent was used as a trigger, whereas no change was observed when tissue factor was used. We found that DX-88 delayed contact activator induced coagulation without affecting tissue factor mediated coagulation. For evaluation of coagulation during DX-88 therapy, the use of PT or tissue factor-activated TEG may be preferable.


A resurgent HIV-1 epidemic among men who have sex with men in the era of potent antiretroviral therapy.
Reducing viral load, highly active antiretroviral therapy has the potential to limit onwards transmission of HIV-1 and thus help contain epidemic spread. However, increases in risk behaviour and resurgent epidemics have been widely reported post-highly active antiretroviral therapy. The aim of this study was to quantify the impact that highly active antiretroviral therapy had on the epidemic. We focus on the HIV-1 epidemic among men who have sex with men in the Netherlands, which has been well documented over the past 20 years within several long-standing national surveillance programs. We used a mathematical model including highly active antiretroviral therapy use and estimated the changes in risk behaviour and diagnosis rate needed to explain annual data on HIV and AIDS diagnoses. We show that the reproduction number R(t), a measure of the state of the epidemic, declined early on from initial values above two and was maintained below one from 1985 to 2000. Since 1996, when highly active antiretroviral therapy became widely used, the risk behaviour rate has increased 66%, resulting in an increase of R(t) to 1.04 in the latest period 2000-2004 (95% confidence interval 0.98-1.09) near or just above the threshold for a self-sustaining epidemic. Hypothetical scenario analysis shows that the epidemiological benefits of highly active antiretroviral therapy and earlier diagnosis on incidence have been entirely offset by increases in the risk behaviour rate. We provide the first detailed quantitative analysis of the HIV epidemic in a well defined population and find a resurgent epidemic in the era of highly active antiretroviral therapy, most likely predominantly caused by increasing sexual risk behaviour.


Comparative electrochemical inactivation of bacteria and bacteriophage.
Electric fields and currents have been shown to be capable of disinfecting drinking water and reducing the numbers of bacteria and yeast in food. However, little research has been conducted regarding the effectiveness of electric fields and currents in the inactivation of viruses. The objective of this study was to compare the ability of bacteria and bacteriophage to survive exposure to direct electric current in an electrochemical cell, where they would be subject to irreversible membrane permeabilization processes, direct oxidation of cellular/viral constituents by electric current, and disinfection by electrochemically generated oxidants. Suspensions of the bacteria Escherichia coli and Pseudomonas aeruginosa and bacteriophage MS2 and PRD1 at both high (approximately 1 x 10(6)CFU or PFU/mL) and low (approximately 1 x 10(3)CFU or PFU/mL) population densities were exposed to currents ranging from 25 to 350 mA in 5s pulses. Post-exposure plaque counts of the bacteriophage were proportionally higher than bacterial culturable counts at corresponding current exposures. E. coli and MS2 were then exposed to 5 mA for 20 min at both high and low population densities. The inactivation rate of E. coli was 2.1-4.3 times greater than that of MS2. Both bacteria and bacteriophage were more resistant to exposure to direct current at higher population densities. Also, amelioration of inactivation within the electrochemical cell by the reducing agent glutathione indicates the major mechanism of inactivation in the electrochemical cell is disinfection by electrochemically generated oxidants. The implications of these results are that technologies relying upon direct current to reduce the numbers of microbes in food and water may not be sufficient to reduce the numbers of potentially pathogenic viruses and ensure the safety of the treated food or water.


Long-term effect of MPTP in the mouse brain in relation to aging: neurochemical and immunocytochemical analysis.
The long-term effect of the parkinsonism-inducing neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) on central monoaminergic neurons in young (2-3 months) and aging (12 months) C57BL/6 mice has been studied using neurochemical and immunocytochemical techniques. MPTP treatment (4 x 20 mg/kg i.p. given 12 h apart) resulted in significant depletion of dopamine (DA) concentration in the striatum, substantia nigra, nucleus accumbens, and olfactory tubercle 1 week after treatment in both young and aging mice. Although a decreased DA concentration in the ventral tegmental area was not seen in young mice, aging mice did show a significant decrease. The extent of decrease of DA concentration was greater in aging mice than in young mice in all areas investigated except in dorsal striatum. The long-term effect of MPTP on DA neurons in young mice included considerable recovery of DA concentration in both nigrostriatal and mesolimbic DA systems following the initial profound depletion; such recovery was minimal in aging mice, even 3 months after MPTP treatment. In young mice treated with MPTP, no significant change of norepinephrine (NE) or serotonin (5-HT) concentration was observed in any area investigated while a significant decrease of NE and 5-HT concentration was seen in several brain areas investigated in aging mice. Immunocytochemical analysis revealed that the MPTP injection resulted in marked disappearance of tyrosine hydroxylase (TH)-immunoreactive (IR) fibers in striatum of both young and aging mice 1 week following treatment. Partial recovery of TH-IR fibers was seen 5 weeks or 3 months after MPTP treatment in young mice, while no such apparent recovery was seen in aging mice. Aging mice also showed significant decrease in the number of TH-positive cell bodies in the substantia nigra and ventral tegmental area through all periods investigated, while such a significant decrease was only seen in the substantia nigra of young mice 1 week after treatment. We conclude that aging mice are more sensitive to MPTP and show more widespread damage to the monoaminergic systems than young mice, suggesting that MPTP-treated aging mice provide a more useful model for studying anatomical and neurochemical characteristics of Parkinson's disease than young mice.


Increased incidence and severity of the systemic inflammatory response syndrome in patients deficient in mannose-binding lectin.
To determine whether pediatric PICU patients with mannose-binding lectin (MBL) gene polymorphisms associated with low levels of the functional protein have an increased risk of developing sepsis and SIRS. A prospective, observational cohort study in a 22-bed PICU in a tertiary referral centre. One hundred consecutive admissions to a PICU with at least one organ system failure longer than 12 h. Patients were classified into those with infectious or non-infectious insults as the primary reason for intensive care admission. Patients were followed to determine which developed sepsis or non-infection related SIRS using standard criteria. Of the 100 patients 50 had infectious and 50 had non-infectious insults as the precipitant for admission. 42 patients had variant MBL alleles (determined by MBL-2 gene exon 1 and promoter polymorphisms) and were significantly over-represented amongst the 59 patients that developed SIRS. This effect was not explained by differences in age, sex or ethnicity and was seen in both the infection and non-infection subgroups. In patients with infection, variant MBL alleles were associated with increased systemic response (2/15 with localised infection, 10/19 with sepsis and 12/16 with septic shock). MBL serum levels showed close concordance with the genotype and indicated that MBL levels less than 1000 ng/ml are associated with a greatly increased risk of SIRS. MBL-2 exon 1 polymorphisms with low serum levels of functional MBL protein are associated with a greatly increased risk of developing SIRS and of progression from infection to sepsis and septic shock in paediatric ICU patients.


T-type Ca2+ channels encode prior neuronal activity as modulated recovery rates.
T-type Ca2+ channels give rise to low-threshold inward currents that are central determinants of neuronal excitability. The availability of T-type Ca2+ channels is strongly influenced by voltage-dependent inactivation and recovery from inactivation. Here, we show that native and cloned T-type Ca2+ channel subunits selectively encode specific aspects of prior membrane potential changes via a powerful modulation of the rates with which these channels recover from inactivation. Increasing the duration of subthreshold (-70 to -55 mV) conditioning depolarizations caused a pronounced slowing of subsequent recovery from inactivation of both cloned (Ca(v)3.1-3.3) and native T-type channels (thalamic neurones). The scaling of recovery rates with increasing duration of conditioning depolarizations could be well described by a power law function. Different T-type channel isoforms exhibited overlapping but complementary ranges of recovery rates. Intriguingly, scaling of recovery rates was dramatically reduced in Ca(v)3.2 and Ca(v)3.3, but not Ca(v)3.1 subunits, when mock action potentials were superimposed on conditioning depolarizations. Our results suggest that different T-type channel subunits exhibit dramatic differences in scaling relationships, in addition to well-described differences in other biophysical properties. Furthermore, the availability of T-type channels is powerfully modulated over time, depending on the patterns of prior activity that these channels have encountered. These data provide a novel mechanism for cellular short-term plasticity on the millisecond to second time scale that relies on biophysical properties of specific T-type Ca2+ channel subunits.


Entinostat induces antitumor immune responses through immune editing of tumor neoantigens.
Although immune-checkpoint inhibitors (ICIs) have been a remarkable advancement in bladder cancer treatment, the response rate to single-agent ICIs remains suboptimal. There has been substantial interest in the use of epigenetic agents to enhance ICI efficacy, although precisely how these agents potentiate ICI response has not been fully elucidated. We identified entinostat, a selective HDAC1/3 inhibitor, as a potent antitumor agent in our immune-competent bladder cancer mouse models (BBN963 and BBN966). We demonstrate that entinostat selectively promoted immune editing of tumor neoantigens, effectively remodeling the tumor immune microenvironment, resulting in a robust antitumor response that was cell autonomous, dependent upon antigen presentation, and associated with increased numbers of neoantigen-specific T cells. Finally, combination treatment with anti-PD-1 and entinostat led to complete responses and conferred long-term immunologic memory. Our work defines a tumor cell-autonomous mechanism of action for entinostat and a strong preclinical rationale for the combined use of entinostat and PD-1 blockade in bladder cancer.


The Feasibility and Acceptability of "Arise": An Online Substance Abuse Relapse Prevention Program.
The purpose of this study was to test the feasibility and acceptability of a novel online adolescent substance abuse relapse prevention tool, "Arise" (3C Institute, Cary, NC). The program uses an innovative platform including interactive instructional segments and skill-building games to help adolescents learn and practice coping skills training strategies. We conducted a pilot test with nine adolescents in substance abuse treatment (44 percent female) and a feasibility test with treatment providers (n=8; 50 percent female). Adolescents interacted with the program via a secure Web site for approximately 30 minutes for each of two instructional units. Treatment providers reviewed the same material at their own pace. All participants completed a questionnaire with items assessing usability, acceptability, understanding, and subjective experience of the program. Regarding feasibility, recruitment of this population within the study constraints proved challenging, but participant retention in the trial was high (no attrition). Adolescents and treatment providers completed the program with no reported problems, and overall we were able to collect data as planned. Regarding acceptability, the program received strong ratings from both adolescents and providers, who found the prototype informative, engaging, and appealing. Both groups strongly recommended continuing development. We were able to deliver the intervention as intended, and acceptability ratings were high, demonstrating the feasibility and acceptability of online delivery of engaging interactive interventions. This study contributes to our understanding of how interactive technologies, including games, can be used to modify behavior in substance abuse treatment and other health areas.


Insulin facilitation of muscle protein synthesis following resistance exercise in hindlimb-suspended rats is independent of a rapamycin-sensitive pathway.
Hindlimb suspension (HS) results in rapid losses of muscle mass, which may in part be explained by attenuated rates of protein synthesis. Mammalian target of rapamycin (mTOR) regulates protein synthesis and has been implicated as a potential mediator of the muscle mass decrement with HS. This study examined the effect of resistance exercise, a muscle hypertrophy stimulant, on rates of protein synthesis after 4 days of HS in mature male Sprague-Dawley rats. Flywheel resistance exercise (2 sets x 25 repetitions) was conducted on days 2 and 4 of HS (HSRE). Sixteen hours after the last exercise bout, soleus muscles were assessed for in vitro rates of protein synthesis, with and without insulin (signaling agonist) and/or rapamycin (mTOR inhibitor). Results demonstrated that soleus mass was reduced (P < 0.05) with HS, but this loss of mass was not observed (P > 0.05) with HSRE. Muscle protein synthesis was diminished (P < 0.05) with HS, with or without insulin. HSRE also had reduced rates of synthesis without insulin; however, insulin administration yielded higher (P < 0.05) rates in HSRE compared with HS or control. Rapamycin diminished protein synthesis in all groups (P < 0.05), but insulin rescued synthesis rates in HS and HSRE to levels similar to insulin alone for each group, suggesting that alternate signaling pathways develop to increase protein synthesis with HS. These results demonstrate that the capacity for an augmented anabolic response to resistance exercise is maintained after 4 days of HS and is independent of a rapamycin-sensitive pathway.


Engineering osteochondral constructs through spatial regulation of endochondral ossification.
Chondrogenically primed bone marrow-derived mesenchymal stem cells (MSCs) have been shown to become hypertrophic and undergo endochondral ossification when implanted in vivo. Modulating this endochondral phenotype may be an attractive approach to engineering the osseous phase of an osteochondral implant. The objective of this study was to engineer an osteochondral tissue by promoting endochondral ossification in one layer of a bilayered construct and stable cartilage in the other. The top half of bilayered agarose hydrogels were seeded with culture expanded chondrocytes (termed the chondral layer) and the bottom half of the bilayered agarose hydrogels with MSCs (termed the osseous layer). Constructs were cultured in chondrogenic medium for 21days and thereafter were either maintained in chondrogenic medium, transferred to hypertrophic medium, or implanted subcutaneously into nude mice. This structured chondrogenic bilayered co-culture was found to enhance chondrogenesis in the chondral layer, appearing to help re-establish the chondrogenic phenotype that is lost in chondrocytes during monolayer expansion. Furthermore, the bilayered co-culture appeared to suppress hypertrophy and mineralization in the osseous layer. The addition of hypertrophic factors to the media was found to induce mineralization of the osseous layer in vitro. A similar result was observed in vivo where endochondral ossification was restricted to the osseous layer of the construct, leading to the development of an osteochondral tissue. This novel approach represents a potential new treatment strategy for the repair of osteochondral defects.


Vertical distraction osteogenesis of edentulous ridges for improvement of oral implant positioning: a clinical report of preliminary results.
This study examined the opportunities offered by intraoral distraction osteogenesis to vertically elongate insufficient alveolar ridges and thereby improve local anatomy for ideal implant placement. Eight patients presenting with vertically deficient edentulous ridges were treated by means of the distraction osteogenesis principle with an intraoral alveolar distractor. Two to 3 months after consolidation of the distracted segments, 26 implants were placed in the distracted areas. Four to 6 months later, abutments were connected and prosthetic loading of the implants was started. The mean follow-up after initial prosthetic loading was 14 months. In all patients, the desired bone gain was reached at the end of distraction (mean vertical bone gain of 8.5 mm). Probing depth, Bleeding Index, and Plaque Index around implants were evaluated, and Periotest values were also calculated. The cumulative success rate of implants was 100%. Radiographic examinations 12 months after functional loading of implants showed a significant increase in the density of the newly generated bone in the distracted areas. This technique seems to be reliable, and the regenerated bone has withstood the functional demands of implant loading. Success rates of implants, periodontal indices of peri-implant soft tissues, and Periotest values were consistent with those reported in the literature regarding implants placed in native bone.


p35 and p39 are essential for cyclin-dependent kinase 5 function during neurodevelopment.
Cyclin-dependent kinase 5 (Cdk5) plays a pivotal role in brain development and neuronal migration. Cdk5 is abundant in postmitotic, terminally differentiated neurons. The ability of Cdk5 to phosphorylate substrates is dependent on activation by its neuronal-specific activators p35 and p39. There exist striking differences in the phenotypic severity of Cdk5-deficient mice and p35-deficient mice. Cdk5-null mutants show a more severe disruption of lamination in the cerebral cortex, hippocampus, and cerebellum. In addition, Cdk5-null mice display perinatal lethality, whereas p35-null mice are viable. These discrepancies have been attributed to the function of other Cdk5 activators, such as p39. To understand the roles of p39 and p35, we created p39-null mice and p35/p39 compound-mutant mice. Interestingly, p39-null mice show no obvious detectable abnormalities, whereas p35(-/-)p39(-/-) double-null mutants are perinatal lethal. We show here that the p35(-/-)p39(-/-) mutants exhibit phenotypes identical to those of the Cdk5-null mutant mice. Other compound-mutant mice with intermediate phenotypes allow us to determine the distinct and redundant functions between p35 and p39. Our data strongly suggest that p35 and p39 are essential for Cdk5 activity during the development of the nervous system. Thus, p35 and p39 are likely to be the principal, if not the only, activators of Cdk5.


Immunocytochemical characterization of the high-affinity thiazide diuretic receptor in rabbit renal cortex.
Thiazide diuretics increase urinary NaCl excretion primarily by inhibiting Na and Cl transport across the apical membrane of cells in the renal distal tubule. Although these diuretics bind to a membrane protein that couples transport of Na and Cl directly, the molecular nature of this transporter and its localization in the mammalian kidney remain controversial. The present experiments were designed to develop monoclonal antibodies to the high-affinity thiazide diuretic receptor to investigate its molecular characteristics and its cellular and subcellular localization in rabbit kidney. Mice were immunized with high-affinity thiazide diuretic receptors that had been partially purified from rabbit kidney cortex. Resulting hybridomas were screened for the ability to immunoprecipitate thiazide diuretic receptors that were labeled with the thiazide-like diuretic [3H]metolazone. A single hybridoma (MAb JM5) produced antibodies capable of immunoprecipitating up to 80% of the labeled thiazide receptors from solubilized renal cortical membranes. MAb JM5 reacted with a 125-kDa protein on Western blots of solubilized renal cortical apical membranes. It stained the apical membrane of cells in the distal convoluted and connecting tubule but did not stain proximal tubules, glomeruli, or interstitial structures. Less intense staining of apical membranes of principal cells in the collecting tubule and a subpopulation of cells in the thick ascending limb were also present. These results indicate that the high-affinity thiazide diuretic receptor comprises a 125-kDa protein that localizes to the apical membrane of cells in the renal distal tubule.


Diagnosis and management of asymptomatic bacteriuria in kidney transplant recipients: a survey of current practice in Europe.
Asymptomatic bacteriuria is frequent in kidney transplant recipients (KTRs). However, there is no consensus on diagnosis or management. We conducted a European survey to explore current practice related to the diagnosis and management of asymptomatic bacteriuria in adult KTRs. A panel of experts from the European Renal Association-European Dialysis Transplant Association/Developing Education Science and Care for Renal Transplantation in European States working group and the European Study Group for Infections in Compromised Hosts of the European Society of Clinical Microbiology and Infectious Diseases designed this cross-sectional, questionnaire-based, self-administered survey. Invitations to participate were e-mailed to European physicians involved in the care of KTRs. Two hundred and forty-four participants from 138 institutions in 25 countries answered the survey (response rate 30%). Most participants [72% (176/244)] said they always screen for asymptomatic bacteriuria in KTRs. Six per cent (15/240) reported never treating asymptomatic bacteriuria with antibiotics. When antimicrobial treatment was used, 24% of the participants (53/224) said they would start with empirical antibiotics. For an episode of asymptomatic bacteriuria caused by a fully susceptible microorganism and despite no contraindications, a majority of participants (121/223) said they would use a fluoroquinolone (n = 56), amoxicillin/clavulanic acid (n = 38) or oral cephalosporins (n = 27). Screening for and treating asymptomatic bacteriuria are common in KTRs despite uncertainties around the benefits and harms. In an era of antimicrobial resistance, further studies are needed to address the diagnosis and management of asymptomatic bacteriuria in these patients.


Stroke incidence and survival among middle-aged adults: 9-year follow-up of the Atherosclerosis Risk in Communities (ARIC) cohort.
Although stroke mortality rates in the United States are well documented, assessment of incidence rates and case fatality are less well studied. A cohort of 15 792 men and women aged 45 to 64 years from a population sample of households in 4 US communities was followed from 1987 to 1995, an average of 7. 2 years. Incident strokes were identified through annual phone contacts and hospital record searching and were then validated. Of the 267 incident definite or probable strokes, 83% (n=221) were categorized as ischemic strokes, 10% (n=27) were intracerebral hemorrhages, and 7% (n=19) were subarachnoid hemorrhages. The age-adjusted incidence rate (per 1000 person-years) of total strokes was highest among black men (4.44), followed by black women (3.10), white men (1.78), and white women (1.24). The black versus white age-adjusted rate ratio (RR) for ischemic stroke was 2.41 (95% CI, 1.85 to 3.15), which was attenuated to 1.38 (95% CI, 1.01 to 1.89) after adjustment for baseline hypertension, diabetes, education level, smoking status, and prevalent coronary heart disease. There was a tendency for the adjusted case fatality rates to be higher among blacks and men, although none of the case fatality comparisons across sex or race was statistically significant. After accounting for established baseline risk factors, blacks still had a 38% greater risk of incident ischemic stroke compared with whites. Identification of new individual and community-level risk factors accounting for the elevated incidence of stroke requires further investigation and incorporation into intervention planning.


The protective effect of berberine against lipopolysaccharide-induced abortion by modulation of inflammatory/immune responses.
Berberine is an isoquinoline derivative alkaloid with anti-inflammatory activity. In this study, we investigated the protective effects of berberine in prevention of LPS-induced abortion. On the gestation day (GD) 9.5, the pregnant mice were injected with low, medium, and high doses of berberine or with PBS. After 4 h, berberine or PBS-pretreated mice were injected with LPS. On GD 11.5, blood samples and uterine tissues were collected from treated mice and percentage of abortion and serum levels of NO, TNF-α, IL-10, and IL12p70 were measured by macroscopic examination and sandwich ELISA, respectively. Our findings show that mice injected with berberine were resistant to LPS-induced abortion. We also found that this treatment prevents the reduction of IL-10 and the enhancement of NO, TNF-α, and IL-12p70 in LPS-treated pregnant mice. Taken together, our results suggest that berberine as an anti-inflammatory agent has protective effects on LPS-induced abortion by modulation of inflammatory/immune responses.


Regional and interspecific variation in Sr, Ca, and Sr/Ca ratios in avian eggshells from the USA.
To examine regional variation in strontium (Sr), which at high concentrations may reduce eggshell quality, increase egg breakage and reproductive failure, we analyzed Sr, and calcium (Ca) concentrations and Sr/Ca ratios in eggshells from 20 avian species from California, Texas, Idaho, Kansas, and Michigan. In addition, we included data previously reported from Arizona to expand the regional comparisons and to better establish patterns of Sr, and Sr/Ca ratios in bird species across the United States. We found Sr concentrations varied significantly among regions, among species, and among foraging guilds; this variability is strongly influenced by the Sr/Ca ratios in surface water from locations close to the region where the eggshells were collected. Sr concentrations and Sr/Ca ratios were significantly higher in bird eggshells from the Volta wildlife region in the San Joaquin Valley, California and in various locales from Arizona. Sr concentrations and Sr/Ca ratios in bird eggshells from other locations in the USA were lower than those detected in these two regions. Among foraging guilds, invertivores had the highest Sr concentrations and Sr/Ca ratios and carnivores had the lowest. In general, the Sr/Ca ratio increased strongly with increasing Sr concentrations (R(2) = 0.99, P < 0.0001). There was a significant correlation (R(2) = 0.58, P < 0.0001) between Sr/Ca ratios in water and the average Sr/Ca ratios in eggshells suggesting that these values could be determined from Sr/Ca ratios in water. Eggshell thickness was poorly correlated with Sr (R(2) = 0.03) but had a significant and positive correlation with Ca and was more properly correlated by a quadratic equation (R(2) = 0.50, Thickness = 2.13 - 0.02Ca - 3.07 * 10(-5)Ca(2)). Our study provides further evidence that Sr accumulates significantly in the avian eggshell, in some regions at concentrations which could be of concern for potential negative effects on reproduction. We suggest that when assessing the effects of metals on avian reproduction in regions with high Sr deposits in rock and soil, Sr concentrations in the eggshell also should be measured to evaluate additional effects on thickness and reproduction.


Combined ultrasound and nerve stimulator-guided deep nerve block may decrease the rate of local anesthetics systemic toxicity: a randomized clinical trial.
Ultrasound guidance might decrease the incidence of local anesthetics systemic toxicity (LAST) for many peripheral nerve blocks compared with nerve stimulator guidance. However, it remains uncertain whether ultrasound guidance is superior to nerve stimulator guidance for deep nerve block of the lower extremity. This study was designed to investigate whether deep nerve block with ultrasound guidance would decrease the incidence of LAST compared with that with nerve stimulator guidance, and to identify associated risk factors of LAST. Three hundred patients undergoing elective lower limb surgery and desiring lumbar plexus blocks (LPBs) and sciatic nerve blocks (SNBs) were enrolled in this study. The patients were randomly assigned to receive LPBs and SNBs with ultrasound guidance (group U), nerve stimulator guidance (group N) or dual guidance (group M). The primary outcome was the incidence of LAST. The secondary outcomes were the number of needle redirection, motor and sensory block onset and nerve distribution restoration time, as well as associated risk factors. There were 18 patients with LAST, including 12 in group U, 4 in group N and 2 in group M. By multiple comparisons among the three groups, we found that the incidence of LAST in group U (12%) was significantly higher than that in group N (4%)(P = 0.037) and group M(2%)(P = 0.006). The OR of LAST with hepatitis B (HBV) infection and the female sex was 3.352 (95% CI,1.233-9.108, P = 0.013) and 9.488 (95% CI,2.142-42.093, P = 0.0004), respectively. Ultrasound guidance, HBV infection and the female sex were risk factors of LAST with LPBs and SNBs. For patients infected with HBV or female patients receiving LPBs and SNBs, we recommended that combined ultrasound and nerve stimulator guidance should be used to improve the safety. This study was approved by the Ethical Committee of the First Affiliated Hospital of Army Medical University. The protocol was registered prospectively with the Chinese Clinical Trial Registry ( ChiCTR-IOR-16008099 ) on March 15, 2016.


Fostering Children's Connection to Nature Through Authentic Situations: The Case of Saving Salamanders at School.
The aim of this paper is to explore how children learn to form new relationships with nature. It draws on a longitudinal case study of children participating in a stewardship project involving the conservation of salamanders during the school day in Stockholm, Sweden. The qualitative method includes two waves of data collection: when a group of 10-year-old children participated in the project (2015) and 2 years after they participated (2017). We conducted 49 interviews with children as well as using participant observations and questionnaires. We found indications that children developed sympathy for salamanders and increased concern and care for nature, and that such relationships persisted 2 years after participation. Our rich qualitative data suggest that whole situations of sufficient unpredictability triggering free exploration of the area, direct sensory contact and significant experiences of interacting with a species were important for children's development of affective relationships with the salamander species and with nature in an open-ended sense. Saving the lives of trapped animals enabled direct sensory interaction, feedback, increased understanding, and development of new skills for dynamically exploring further ways of saving species in an interactive process experienced as deeply meaningful, enjoyable and connecting. The behavioral setting instilled a sense of pride and commitment, and the high degree of responsibility given to the children while exploring the habitat during authentic situations enriched children's enjoyment. The study has implications for the design of education programs that aim to connect children with nature and for a child-sensitive urban policy that supports authentic nature situations in close spatial proximity to preschools and schools.


Germinal center B cell depletion diminishes CD4+ follicular T helper cells in autoimmune mice.
Continuous support from follicular CD4(+) T helper (Tfh) cells drives germinal center (GC) responses, which last for several weeks to produce high affinity memory B cells and plasma cells. In autoimmune Sle1 and NZB/W F1 mice, elevated numbers of Tfh cells persist, promoting the expansion of self-reactive B cells. Expansion of circulating Tfh like cells have also been described in several autoimmune diseases. Although, the signals required for Tfh differentiation have now been well described, the mechanisms that sustain the maintenance of fully differentiated Tfh are less understood. Recent data demonstrate a role for GC B cells for Tfh maintenance after protein immunization. Given the pathogenic role Tfh play in autoimmune disease, we explored whether B cells are required for maintenance of autoreactive Tfh. Our data suggest that the number of mature autoreactive Tfh cells is controlled by GC B cells. Depletion of B cells in Sle1 autoimmune mice leads to a dramatic reduction in Tfh cells. In NZB/W F1 autoimmune mice, similar to the SRBC immunization model, GC B cells support the maintenance of mature Tfh, which is dependent mainly on ICOS. The CD28-associated pathway is dispensable for Tfh maintenance in SRBC immunized mice, but is required in the spontaneous NZB/W F1 model. These data suggest that mature Tfh cells require signals from GC B cells to sustain their optimal numbers and function in both autoimmune and immunization models. Thus, immunotherapies targeting B cells in autoimmune disease may affect pathogenic Tfh cells.


Open reduction and internal fixation of capitellar fractures with headless screws.
The outcome of operatively treated capitellar fractures has not been reported frequently. The purpose of the present study was to evaluate the clinical, radiographic, and functional outcomes following open reduction and internal fixation of capitellar fractures that were treated with a uniform surgical approach in order to further define the impact on the outcome of fracture type and concomitant lateral column osseous and/or ligamentous injuries. A retrospective evaluation of the upper extremity database at our institution identified sixteen skeletally mature patients (mean age, 40 +/- 17 years) with a closed capitellar fracture. In all cases, an extensile lateral exposure and articular fixation with buried cannulated variable-pitch headless compression screws was performed at a mean of ten days after the injury. Clinical, radiographic, and elbow-specific outcomes, including the Mayo Elbow Performance Index, were evaluated at a mean of 27 +/- 19 months postoperatively. Six Type-I, two Type-III, and eight Type-IV fractures were identified with use of the Bryan and Morrey classification system. Four of five ipsilateral radial head fractures occurred in association with a Type-IV fracture. The lateral collateral ligament was intact in fifteen of the sixteen elbows. Metaphyseal comminution was observed in association with five fractures (including four Type-IV fractures and one Type-III fracture). Supplemental mini-fragment screws were used for four of eight Type-IV fractures and one of two Type-III fractures. All fractures healed, and no elbow had instability or weakness. Overall, the mean ulnohumeral motion was 123 degrees (range, 70 degrees to 150 degrees). Fourteen of the sixteen patients achieved a functional arc of elbow motion, and all patients had full forearm rotation. The mean Mayo Elbow Performance Index score was 92 +/- 10 points, with nine excellent results, six good results, and one fair result. Patients with a Type-IV fracture had a greater magnitude of flexion contracture (p = 0.04), reduced terminal flexion (p = 0.02), and a reduced net ulnohumeral arc (p = 0.01). An ipsilateral radial head fracture did not appear to affect ulnohumeral motion or the functional outcome. Despite the presence of greater flexion contractures at the time of follow-up in elbows with Type-IV fractures or fractures with an ipsilateral radial head fracture, good to excellent outcomes with functional ulnohumeral motion can be achieved following internal fixation of these complex fractures. Type-IV injuries may be more common than previously thought; such fractures often are associated with metaphyseal comminution or a radial head fracture and may require supplemental fixation.


A case-control study of risk factors for hepatocellular carcinoma in Henan, China.
A case-control study was carried out to investigate risk factors in the development of hepatocellular carcinoma (HCC) in Henan, China. A total of 152 patients with HCC and 115 control patients with nonhepatic disease were included in this study. The risk factor analysis indicated that individual history of liver disease (odds ratio [OR]=11.77; 95% confidence interval [CI]=3.94-35.18), family history of liver disease (OR=11.80, 95% CI=2.75-50.61), drinking of alcohol (OR=1.06, 95% CI=1.02-1.10), corn consumption (OR=19.43, 95% CI=3.67-102.98), peanut and peanut oil consumption (OR=13.75, 95% CI=3.69-51.16), and infection with hepatitis B virus (HBV) (OR=44.59, 95% CI=12.54-158.49) were significantly different between the case and control groups (P < 0.01). We also found that there was a dose-response relationship between drinking of alcohol and development of HCC, whereas heavy drinkers experienced an approximately 3-4-fold risk increase compared with nondrinkers. In further analysis, a 1:1 pair-matched case-control study was performed. One hundred thirteen of 152 HCC patients were randomly selected to be pair-matched by sex and age (+/-5 years) with the controls with nonhepatic disease. The results from the 1:1 pair-matched case-control study were consistent with the results from the group-matched case-control study. The findings of this study suggest that hepatitis virus (HBV, HCV) infection, drinking of alcohol, and dietary exposure to aflatoxin are likely etiologic agents of HCC in Henan, China.


Military Medicine Interest Groups in U.S. Medical Schools.
Medical student interest groups are organizations that help expose students to different medical specialties and fields of medicine while in medical school. Military medicine interest groups (MMIGs) are a particular type of interest group that spreads information about military medicine, fosters mentorship, and camaraderie between students and military faculty, and increases the opportunities for leadership while in medical school. Surveys were sent to all U.S. medical schools to determine how many schools had an MMIG. If a medical school had a group, a second survey was sent to the student leader to determine more information about how their group operated (such as type of participants, funding sources, activities, faculty involvement, military health care provider involvement, etc.). Fifty-six percent of U.S. medical schools who responded were found to have an MMIG and most participants were students in the Health Professions Scholarship Program. Information about military medicine was found to be the biggest impact of having a group at a medical school and student leaders expressed they wished to have more military health care provider involvement. The results of this study could help start MMIGs at other medical schools, as well as give ideas to current MMIGs on how other groups operate.


Phase I/II trial of metronomic chemotherapy with daily dalteparin and cyclophosphamide, twice-weekly methotrexate, and daily prednisone as therapy for metastatic breast cancer using vascular endothelial growth factor and soluble vascular endothelial growth factor receptor levels as markers of response.
PURPOSE Preclinical studies indicate that metronomic chemotherapy is antiangiogenic and synergistic with other antiangiogenic agents. We designed a phase I/II study to evaluate the safety and activity of adding dalteparin and prednisone to metronomic cyclophosphamide and methotrexate in women with measurable metastatic breast cancer (MBC). PATIENTS AND METHODS Patients received daily dalteparin and oral cyclophosphamide, twice-weekly methotrexate, and daily prednisone (dalCMP). The primary study end point was clinical benefit rate (CBR), a combination of complete response (CR), partial response (PR), and prolonged stable disease for > or = 24 weeks (pSD). Secondary end points included time to progression (TTP), duration of response, and overall survival (OS). Biomarker response to treatment was assessed by using plasma vascular endothelial growth factor (VEGF) and soluble VEGF receptors (sVEGFRs) -1 and -2. Results Forty-one eligible patients were accrued. Sixteen (39%) had no prior chemotherapy for MBC; 15 (37%) had two or more chemotherapy regimens for MBC. Toxicities were minimal except for transient grade 3 elevation of liver transaminases in 11 patients (27%) and grade 3 vomiting in one patient (2%). One patient (2%) had CR, six (15%) had PR, and three (7%) had pSD, for a CBR of 10 (24%) of 41 patients. Median TTP was 10 weeks (95% CI, 8 to 17 weeks), and median OS was 48 weeks (95% CI, 32 to 79 weeks). VEGF levels decreased but not significantly, whereas sVEGFR-1 and -2 levels increased significantly after 2 weeks of therapy. There was no correlation between response and VEGF, sVEGFR-1, or sVEGFR-2 levels. CONCLUSION Metronomic dalCMP is safe, well tolerated, and clinically active in MBC.


NAADP, cADPR and IP3 all release Ca2+ from the endoplasmic reticulum and an acidic store in the secretory granule area.
Inositol trisphosphate and cyclic ADP-ribose release Ca2+ from the endoplasmic reticulum via inositol trisphosphate and ryanodine receptors, respectively. By contrast, nicotinic acid adenine dinucleotide phosphate may activate a novel Ca2+ channel in an acid compartment. We show, in two-photon permeabilized pancreatic acinar cells, that the three messengers tested could each release Ca2+ from the endoplasmic reticulum and also from an acid store in the granular region. The nicotinic acid adenine dinucleotide phosphate action on both types of store, like that of cyclic ADP-ribose but unlike inositol trisphosphate, depended on operational ryanodine receptors, since it was blocked by ryanodine or ruthenium red. The acid Ca2+ store in the granular region did not have Golgi or lysosomal characteristics and might therefore be associated with the secretory granules. The endoplasmic reticulum is predominantly basal, but thin extensions penetrate into the granular area and cytosolic Ca2+ signals probably initiate at sites where endoplasmic reticulum elements and granules come close together.


Hypertonic dextrose injections (prolotherapy) in the treatment of symptomatic knee osteoarthritis: A systematic review and meta-analysis.
Hypertonic dextrose injections (prolotherapy) is an emerging treatment for symptomatic knee osteoarthritis (OA) but its efficacy is uncertain. We conducted a systematic review with meta-analysis to synthesize clinical evidence on the effect of prolotherapy for knee OA. Fifteen electronic databases were searched from their inception to September 2015. The primary outcome of interest was score change on the Western Ontario and McMaster Universities Arthritis Index (WOMAC). Three randomized controlled trials (RCTs) of moderate risk of bias and one quasi-randomized trial were included, with data from a total of 258 patients. In the meta-analysis of two eligible studies, prolotherapy is superior to exercise alone by a standardized mean difference (SMD) of 0.81 (95% CI: 0.18 to 1.45, p = 0.012), 0.78 (95% CI: 0.25 to 1.30, p = 0.001) and 0.62 (95% CI: 0.04 to 1.20, p = 0.035) on the WOMAC composite scale; and WOMAC function and pain subscale scores respectively. Moderate heterogeneity exists in all cases. Overall, prolotherapy conferred a positive and significant beneficial effect in the treatment of knee OA. Adequately powered, longer-term trials with uniform end points are needed to better elucidate the efficacy of prolotherapy.


Single Molecule Localization and Discrimination of DNA-Protein Complexes by Controlled Translocation Through Nanocapillaries.
Through the use of optical tweezers we performed controlled translocations of DNA-protein complexes through nanocapillaries. We used RNA polymerase (RNAP) with two binding sites on a 7.2 kbp DNA fragment and a dCas9 protein tailored to have five binding sites on λ-DNA (48.5 kbp). Measured localization of binding sites showed a shift from the expected positions on the DNA that we explained using both analytical fitting and a stochastic model. From the measured force versus stage curves we extracted the nonequilibrium work done during the translocation of a DNA-protein complex and used it to obtain an estimate of the effective charge of the complex. In combination with conductivity measurements, we provided a proof of concept for discrimination between different DNA-protein complexes simultaneous to the localization of their binding sites.


Genotype tunes pancreatic ductal adenocarcinoma tissue tension to induce matricellular fibrosis and tumor progression.
Fibrosis compromises pancreatic ductal carcinoma (PDAC) treatment and contributes to patient mortality, yet antistromal therapies are controversial. We found that human PDACs with impaired epithelial transforming growth factor-β (TGF-β) signaling have high epithelial STAT3 activity and develop stiff, matricellular-enriched fibrosis associated with high epithelial tension and shorter patient survival. In several KRAS-driven mouse models, both the loss of TGF-β signaling and elevated β1-integrin mechanosignaling engaged a positive feedback loop whereby STAT3 signaling promotes tumor progression by increasing matricellular fibrosis and tissue tension. In contrast, epithelial STAT3 ablation attenuated tumor progression by reducing the stromal stiffening and epithelial contractility induced by loss of TGF-β signaling. In PDAC patient biopsies, higher matricellular protein and activated STAT3 were associated with SMAD4 mutation and shorter survival. The findings implicate epithelial tension and matricellular fibrosis in the aggressiveness of SMAD4 mutant pancreatic tumors and highlight STAT3 and mechanics as key drivers of this phenotype.


An exploratory ecotoxicity study of primary microplastics versus aged in natural waters and wastewaters.
Current understanding of how environmental aging of microplastics contributes to their ecotoxicity is low. We investigated whether incubation of microplastics in waters with different organic load and toxic potential alters the toxicity of microplastics to crustacean Daphnia magna, fish embryos Danio rerio and plant Lemna minor. Polyethylene primary microplastics; specifically microbeads from facial scrub; were subjected to 3-weeks incubation in low affected spring water, river water, effluent from the municipal wastewater treatment plant (WWTP) and municipal landfill leachate. Primary microplastics had no acute effect on D. magna mobility and D. rerio embryos development. While high organic load wastewaters; WWTP effluent and landfill leachate; showed evident toxicity for D. magna and D. rerio embryos, microplastics aged in these wastewaters had no effect. This suggests that adsorption of pollutants from wastewaters to microplastic particles was not high enough to induce acute toxicity to D. magna and D. rerio. On the contrary, primary microplastics affected the root growth of L. minor. Interestingly, aging of microplastics in low organic-load waters mitigated the toxicity of microplastics for L. minor, while microplastics aged in high-organic load waters had the same adverse effect as primary microplastics. Partly, these effects can be explained by different extent of coating on microplastics in different water samples. This study suggests that aging of microplastics in wastewaters and natural waters did not significantly enhance the toxicity to selected test species, but further studies on plants may be of interest.


Adaptive divergence in experimental populations of Pseudomonas fluorescens. V. Insight into the niche specialist fuzzy spreader compels revision of the model Pseudomonas radiation.
Pseudomonas fluorescens is a model for the study of adaptive radiation. When propagated in a spatially structured environment, the bacterium rapidly diversifies into a range of niche specialist genotypes. Here we present a genetic dissection and phenotypic characterization of the fuzzy spreader (FS) morphotype-a type that arises repeatedly during the course of the P. fluorescens radiation and appears to colonize the bottom of static broth microcosms. The causal mutation is located within gene fuzY (pflu0478)-the fourth gene of the five-gene fuzVWXYZ operon. fuzY encodes a β-glycosyltransferase that is predicted to modify lipopolysaccharide (LPS) O antigens. The effect of the mutation is to cause cell flocculation. Analysis of 92 independent FS genotypes showed each to have arisen as the result of a loss-of-function mutation in fuzY, although different mutations have subtly different phenotypic and fitness effects. Mutations within fuzY were previously shown to suppress the phenotype of mat-forming wrinkly spreader (WS) types. This prompted a reinvestigation of FS niche preference. Time-lapse photography showed that FS colonizes the meniscus of broth microcosms, forming cellular rafts that, being too flimsy to form a mat, collapse to the vial bottom and then repeatably reform only to collapse. This led to a reassessment of the ecology of the P. fluorescens radiation. Finally, we show that ecological interactions between the three dominant emergent types (smooth, WS, and FS), combined with the interdependence of FS and WS on fuzY, can, at least in part, underpin an evolutionary arms race with bacteriophage SBW25Φ2, to which mutation in fuzY confers resistance.


Rail Track Detection and Projection-Based 3D Modeling from UAV Point Cloud.
The expansion of the railway industry has increased the demand for the three-dimensional modeling of railway tracks. Due to the increasing development of UAV technology and its application advantages, in this research, the detection and 3D modeling of rail tracks are investigated using dense point clouds obtained from UAV images. Accordingly, a projection-based approach based on the overall direction of the rail track is proposed in order to generate a 3D model of the railway. In order to extract the railway lines, the height jump of points is evaluated in the neighborhood to select the candidate points of rail tracks. Then, using the RANSAC algorithm, line fitting on these candidate points is performed, and the final points related to the rail are identified. In the next step, the pre-specified rail piece model is fitted to the rail points through a projection-based process, and the orientation parameters of the model are determined. These parameters are later improved by fitting the Fourier curve, and finally a continuous 3D model for all of the rail tracks is created. The geometric distance of the final model from rail points is calculated in order to evaluate the modeling accuracy. Moreover, the performance of the proposed method is compared with another approach. A median distance of about 3 cm between the produced model and corresponding point cloud proves the high quality of the proposed 3D modeling algorithm in this study.


Analytical description of a Fabry-Perot spectrometer. 8: Optimum operation with equidistant equal-noise sampling.
The optimum parameters of operation for a Fabry-Perot spectrometer used in the equidistant equal-noise sampling method have been calculated. The results, expressed in terms of normalized halfwidths at half-height, are: etalon (a*) = 0.078 (0.095), linewidth (dg*) = 0.13 (0.19), and aperture (f*) = 0.10 (0.19) for temperature (wind) determinations. The etalon widths correspond to rather low reflectivities, namely, 0.62 and 0.56. The critical number of samples, required for unambiguous determinations of a measured profile, are found to be equal to 8 and 12. The usefulness of the equal-noise method in absorption measurements is discussed.


Rho GTPase isoforms in cell motility: Don't fret, we have FRET.
The Rho-family of p21 small GTPases are directly linked to the regulation of actin-based motile machinery and play a key role in the control of cell migration. Aside from the original and most well-characterized canonical Rho GTPases RhoA, Rac1, and Cdc42, numerous isoforms of these key proteins have been identified and shown to have specific roles in regulating various cellular motility processes. The major difficulty in addressing these isoform-specific effects is that isoforms typically contain highly similar primary amino acid sequences and thus are able to interact with the same upstream regulators and the downstream effector targets. Here, we will introduce the major members of each GTPase subfamily and discuss recent advances in the design and application of fluorescent resonance energy transfer-based probes, which are at the forefront of the technologies available to directly probe the differential, spatiotemporal activation dynamics of these proteins in live single cells. Currently, it is possible to specifically detect the activation status of RhoA vs. RhoC isoforms, as well as Cdc42 vs. TC-10 isoforms in living cells. Clearly, additional efforts are still required to produce biosensor systems capable of detecting other isoforms of Rho GTPases including RhoB, Rac2/3, RhoG, etc. Through such efforts, we will uncover the isoform-specific roles of these near-identical proteins in living cells, clearly an important area of the Rho GTPase biology that is not yet fully appreciated.


SEPT7-mediated regulation of Ca2+ entry through Orai channels requires other septin subunits.
Orai channels are plasma membrane resident Ca2+ channels that allow extracellular Ca2+ uptake after depletion of ER-Ca2+ stores by a process called store-operated Ca2+ entry (SOCE). Septins of the SEPT2 subgroup act as positive regulators of SOCE in human nonexcitable cells. SEPT2 subgroup septins form the central core of hetero-hexameric or hetero-octameric complexes with SEPT6, SEPT7 and SEPT9 subgroup septins. The presence of fewer septin encoding genes coupled with ease of genetic manipulation allows for better understanding of septin subgroup function in Drosophila. Our earlier findings show that although dSEPT7 reduction does not alter Orai-mediated Ca2+ entry during SOCE, it results in constitutive activation of Orai channels in resting neurons. Here, we have investigated the role of other septin subgroup members in regulating Orai channel activation in Drosophila neurons by both cellular and functional assays. We show that dSEPT1, a SEPT2 subgroup septin can exist in a complex with dSEPT2 and dSEPT7 in the central nervous system (CNS) of Drosophila. Our findings suggest that the nature of septin filaments and heteromers obtained after reducing septins of different subgroups alters their ability to regulate Orai channel opening. The molecular mechanisms underlying this complex regulation of Orai function by septins require further cellular investigations.


Shape Transformations of Epithelial Shells.
Regulated deformations of epithelial sheets are frequently foreshadowed by patterning of their mechanical properties. The connection between patterns of cell properties and the emerging tissue deformations is studied in multiple experimental systems, but the general principles remain poorly understood. For instance, it is in general unclear what determines the direction in which the patterned sheet is going to bend and whether the resulting shape transformation will be discontinuous or smooth. Here these questions are explored computationally, using vertex models of epithelial shells assembled from prismlike cells. In response to rings and patches of apical cell contractility, model epithelia smoothly deform into invaginated or evaginated shapes similar to those observed in embryos and tissue organoids. Most of the observed effects can be captured by a simpler model with polygonal cells, modified to include the effects of the apicobasal polarity and natural curvature of epithelia. Our models can be readily extended to include the effects of multiple constraints and used to describe a wide range of morphogenetic processes.


Characterization of a recombinant canine coronavirus with a distinct receptor-binding (S1) domain.
Canine alphacoronaviruses (CCoV) exist in two serotypes, type I and II, both of which can cause severe gastroenteritis. Here, we characterize a canine alphacoronavirus, designated CCoV-A76, first isolated in 1976. Serological studies show that CCoV-A76 is distinct from other CCoVs, such as the prototype CCoV-1-71. Efficient replication of CCoV-A76 is restricted to canine cell lines, in contrast to the prototypical type II strain CCoV-1-71 that more efficiently replicates in feline cells. CCoV-A76 can use canine aminopeptidase N (cAPN) receptor for infection of cells, but was unable to use feline APN (fAPN). In contrast, CCoV-1-71 can utilize both. Genomic analysis shows that CCoV-A76 possesses a distinct spike, which is the result of a recombination between type I and type II CCoV, that occurred between the N- and C-terminal domains (NTD and C-domain) of the S1 subunit. These data suggest that CCoV-A76 represents a recombinant coronavirus form, with distinct host cell tropism.


Systems Based Study of the Therapeutic Potential of Small Charged Molecules for the Inhibition of IL-1 Mediated Cartilage Degradation.
Inflammatory cytokines are key drivers of cartilage degradation in post-traumatic osteoarthritis. Cartilage degradation mediated by these inflammatory cytokines has been extensively investigated using in vitro experimental systems. Based on one such study, we have developed a computational model to quantitatively assess the impact of charged small molecules intended to inhibit IL-1 mediated cartilage degradation. We primarily focus on the simplest possible computational model of small molecular interaction with the IL-1 system-direct binding of the small molecule to the active site on the IL-1 molecule itself. We first use the model to explore the uptake and release kinetics of the small molecule inhibitor by cartilage tissue. Our results show that negatively charged small molecules are excluded from the negatively charged cartilage tissue and have uptake kinetics in the order of hours. In contrast, the positively charged small molecules are drawn into the cartilage with uptake and release timescales ranging from hours to days. Using our calibrated computational model, we subsequently explore the effect of small molecule charge and binding constant on the rate of cartilage degradation. The results from this analysis indicate that the small molecules are most effective in inhibiting cartilage degradation if they are either positively charged and/or bind strongly to IL-1α, or both. Furthermore, our results showed that the cartilage structural homeostasis can be restored by the small molecule if administered within six days following initial tissue exposure to IL-1α. We finally extended the scope of the computational model by simulating the competitive inhibition of cartilage degradation by the small molecule. Results from this model show that small molecules are more efficient in inhibiting cartilage degradation by binding directly to IL-1α rather than binding to IL-1α receptors. The results from this study can be used as a template for the design and development of more pharmacologically effective osteoarthritis drugs, and to investigate possible therapeutic options.


T-cadherin is present on endothelial microparticles and is elevated in plasma in early atherosclerosis.
The presence of endothelial cell (EC)-derived surface molecules in the circulation is among hallmarks of endothelial activation and damage in vivo. Previous investigations suggest that upregulation of T-cadherin (T-cad) on the surface of ECs may be a characteristic marker of EC activation and stress. We investigated whether T-cad might also be shed from ECs and in amounts reflecting the extent of activation or damage. Immunoblotting showed the presence of T-cad protein in the culture medium from normal proliferating ECs and higher levels in the medium from stressed/apoptotic ECs. Release of T-cad into the circulation occurs in vivo and in association with endothelial dysfunction. Sandwich ELISA revealed negligible T-cad protein in the plasma of healthy volunteers (0.90 ± 0.90 ng/mL, n = 30), and increased levels in the plasma from patients with non-significant atherosclerosis (9.23 ± 2.61 ng/mL, n = 63) and patients with chronic coronary artery disease (6.93 ± 1.31 ng/mL, n = 162). In both patient groups there was a significant (P = 0.043) dependency of T-cad and degree of endothelial dysfunction as measured by reactive hyperaemia peripheral tonometry. Flow cytometry analysis showed that the major fraction of T-cad was released into the EC culture medium and the plasma as a surface component of EC-derived annexin V- and CD144/CD31-positive microparticles (MPs). Gain-of-function and loss-of-function studies demonstrate that MP-bound T-cad induced Akt phosphorylation and activated angiogenic behaviour in target ECs via homophilic-based interactions. Our findings reveal a novel mechanism of T-cad-dependent signalling in the vascular endothelium. We identify T-cad as an endothelial MP antigen in vivo and demonstrate that its level in plasma is increased in early atherosclerosis and correlates with endothelial dysfunction.


Pediatric Bone Mineral Accrual Z-Score Calculation Equations and Their Application in Childhood Disease.
Annual gains in BMC and areal bone mineral density (aBMD) in children vary with age, pubertal status, height-velocity, and lean body mass accrual (LBM velocity). Evaluating bone accrual in children with bone health-threatening conditions requires consideration of these determinants. The objective of this study was to develop prediction equations for calculating BMC/aBMD velocity SD scores (velocity-Z) and to evaluate bone accrual in youth with health conditions. Bone and body compositions via DXA were obtained for up to six annual intervals in healthy youth (n = 2014) enrolled in the Bone Mineral Density in Childhood Study (BMDCS) . Longitudinal statistical methods were used to develop sex- and pubertal-status-specific reference equations for calculating velocity-Z for total body less head-BMC and lumbar spine (LS), total hip (TotHip), femoral neck, and 1/3-radius aBMD. Equations accounted for (1) height velocity, (2) height velocity and weight velocity, or (3) height velocity and LBM velocity. These equations were then applied to observational, single-center, 12-month longitudinal data from youth with cystic fibrosis (CF; n = 65), acute lymphoblastic leukemia (ALL) survivors (n = 45), or Crohn disease (CD) initiating infliximab (n = 72). Associations between BMC/aBMD-Z change (conventional pediatric bone health monitoring method) and BMC/aBMD velocity-Z were assessed. The BMC/aBMD velocity-Z for CF, ALL, and CD was compared with BMDCS. Annual changes in the BMC/aBMD-Z and the BMC/aBMD velocity-Z were strongly correlated, but not equivalent; LS aBMD-Z = 1 equated with LS aBMD velocity-Z = -3. In CF, BMC/aBMD velocity-Z was normal. In posttherapy ALL, BMC/aBMD velocity-Z was increased, particularly at TotHip (1.01 [-.047; 1.7], p < 0.0001). In CD, BMC/aBMD velocity-Z was increased at all skeletal sites. LBM-velocity adjustment attenuated these increases (eg, TotHip aBMD velocity-Z: 1.13 [0.004; 2.34] versus 1.52 [0.3; 2.85], p < 0.0001). Methods for quantifying the BMC/aBMD velocity that account for maturation and body composition changes provide a framework for evaluating childhood bone accretion and may provide insight into mechanisms contributing to altered accrual in chronic childhood conditions. © 2018 American Society for Bone and Mineral Research.


Laser Fluorescence Illuminates the Soft Tissue and Life Habits of the Early Cretaceous Bird Confuciusornis.
In this paper we report the discovery of non-plumage soft tissues in Confuciusornis, a basal beaked bird from the Early Cretaceous Jehol Biota in northeastern China. Various soft tissues are visualized and interpreted through the use of laser-stimulated fluorescence, providing much novel anatomical information about this early bird, specifically reticulate scales covering the feet, and the well-developed and robust pro- and postpatagium. We also include a direct comparison between the forelimb soft tissues of Confuciusornis and modern avian patagia. Furthermore, apparently large, fleshy phalangeal pads are preserved on the feet. The reticulate scales, robust phalangeal pads as well as the highly recurved pedal claws strongly support Confuciusornis as an arboreal bird. Reticulate scales are more rounded than scutate scales and do not overlap, thus allowing for more flexibility in the toe. The extent of the pro- and postpatagium and the robust primary feather rachises are evidence that Confuciusornis was capable of powered flight, contrary to previous reports suggesting otherwise. A unique avian wing shape is also reconstructed based on plumage preserved. These soft tissues combined indicate an arboreal bird with the capacity for short-term (non-migratory) flight, and suggest that, although primitive, Confuciusornis already possessed many relatively advanced avian anatomical characteristics.


Prevalence of depression after stroke: the Perth Community Stroke Study.
The Perth Community Stroke Study (PCSS) was a population-based study of the incidence, cause, and outcome of acute stroke. Subjects from the study were assessed initially, by examination and interview, and at four- and 12-month follow-ups to determine differences in prevalence of depression between the sexes and between patients with first-ever and recurrent strokes. The prevalence of depressive illness four months after stroke in 294 patients from the PCSS was 23% (18-28%), 15% (11-19%) major depression and 8% (5-11%) minor depression. There were no significant differences between the sexes or between patients with first-ever and recurrent strokes. With a non-hierarchic approach to diagnosis of those with depression, 26% of men and 39% of women had an associated anxiety disorder, mainly agoraphobia. Nine per cent of male and 13% of female patients interviewed had evidence of depression at the time of the stroke. Twelve months after stroke 56% of the men were still depressed (40% major and 16% minor), as were 30% of the women (12% major and 18% minor). The prevalence of depression after stroke was comparable with that reported from other studies, and considerably less than that reported from in-patient and rehabilitation units.


Zinc intakes and plasma concentrations in men with osteoporosis: the Rancho Bernardo Study.
Low zinc intakes and reduced blood zinc concentrations have been reported to be associated with osteoporosis in women. The objective was to examine the independent association between dietary zinc and plasma zinc and the association of each with bone mineral density (BMD) and 4-y bone loss in community-dwelling older men. Of the original Rancho Bernardo Study subjects, 396 men (age: 45-92 y) completed BMD measurements at baseline in 1988-1992 and 4 y later. Osteoporosis was defined as a BMD > or = 2.5 SDs below the mean for young women (a T-score < or = -2.5). At baseline, dietary intake data were collected by using a standard food-frequency questionnaire, and plasma zinc concentrations were measured by using inductively coupled plasma spectroscopy. The mean dietary zinc intake was 11.2 mg, and the mean plasma zinc concentration was 12.7 micromol/L. Plasma zinc was correlated with total zinc intake (diet plus supplements). Dietary zinc intake and plasma zinc concentrations were lower in men with osteoporosis at the hip and spine than in men without osteoporosis at those locations. BMDs for the hip, spine, and distal wrist were significantly lower in men in the lowest plasma zinc quartile (<11.3 micromol/L) than in men with higher plasma zinc concentrations. The association between plasma zinc and BMD was cross-sectional, longitudinal, and independent of age or body mass index. However, plasma zinc did not predict bone loss during the 4-y interval. Dietary zinc intake and plasma zinc each have a positive association with BMD in men.


Determination of HER2 phosphorylation at tyrosine 1221/1222 improves prediction of poor survival for breast cancer patients with hormone receptor-positive tumors.
High expression of total HER2 protein confers poor prognosis for breast cancer patients. HER2 is a member of the HER family consisting of four receptors, HER1 to HER4. HER receptor activity is regulated by a variety of mechanisms, and phosphorylation of the C-terminal part of the HER receptors is a marker for active signaling. The importance of phosphorylation and thereby activation of the HER1 to HER4 receptors, however, has not been investigated concomitantly in breast tumors. In the present study we examined the importance of active HER signaling in breast tumor biopsies and paired metastases, by evaluating the expression of phosphorylated HER1, HER2, HER3, Erk, Akt and the total level of HER4 and HER2. Immunohistochemical analysis was performed on 268 primary breast tumors and 30 paired metastatic lesions from postmenopausal women with hormone receptor-positive breast tumors, who had received adjuvant tamoxifen therapy. The observed protein expression levels were analyzed for co-expression, for correlation to clinicopathological parameters and for prognostic value in relation to disease-free survival and overall survival. Lastly, the difference between protein levels in primary tumors versus metastasis was evaluated. In the primary tumors, 8%, 18%, 14% and 15% of cases were scored positive for total HER2, pHER1, pHER2 and pHER3 expression, respectively. HER4 was expressed with strong intensity in 68% and at moderate intensity in 29% of cases. The activated forms of Akt and Erk were quite uniformly expressed in the categories; negative, moderate or strong. In univariate analysis, expression of total HER2, pHER1, pHER2 and pHER3 was significantly associated with poor disease-free survival. Strong HER4 expression was associated with prolonged disease-free as well as with overall survival. Expression of pAkt and pErk was not correlated with survival. In multivariate analysis, pHER2 expression was clearly an independent marker for poor disease-free survival and overall survival when tested against tumor size, tumor grade, nodal status and HER2. Lastly, comparison of HER receptor expression in metastatic versus primary tumors showed a significant increase in expression of pHER1 and pHER3 in the metastases. In hormone receptor-positive breast cancer, determination of pHER2 yields additional prognostic information about poor prognosis compared with the current clinical standard for measuring HER2.


Transitions to material efficiency in the UK steel economy.
Steel production is energy intensive so already has achieved impressive levels of energy efficiency. If the emissions associated with steel must be reduced in line with the requirements of the UK Climate Change Act, demand for new steel must be reduced. The strategies of 'material efficiency' aim to achieve such a reduction, while delivering the same final services. To meet the emissions targets set into UK law, UK consumption of steel must be reduced to 30 per cent of present levels by 2050. Previous work has revealed six strategies that could contribute to this target, and this paper presents an approximate analysis of the required transition. A macro-economic analysis of steel in the UK shows that while the steel industry is relatively small, the construction and manufacturing sectors are large, and it would be politically unacceptable to pursue options that lead to a major contraction in other sectors. Alternative business models are therefore required, and these are explored through four representative products--one for each final sector with particular emphasis given to options for reducing product weight, and extending product life. Preliminary evidence on the triggers that would lead to customers preferring these options is presented and organized in order to predict required policy measures. The estimated analysis of transitions explored in this paper is used to define target questions for future research in the area.


Effects of acute stress on cognition in older versus younger adults.
Does acute stress differentially alter cognitive functioning in older versus younger adults? While older adults may be better at handling stress psychologically, their physiological systems are less elastic, potentially impairing the cognitive functioning of older adults after a stressor. We examined cognition following an acute stressor among older (n = 65; ages 60-79) and younger (n = 61; ages 25-40) adults. Participants were randomized to complete the Trier Social Stress Test (TSST) in one of three conditions: (a) negative feedback, (b) positive feedback, or (c) no feedback. Participants reported mood states and appraisals of the speech task and we measured cortisol via saliva throughout the study. After the TSST, participants completed standard cognitive tasks to evaluate cognitive flexibility, problem solving, and short-term memory. Results showed that after the TSST, older adults took longer to solve problems compared with younger adults, though they were able to solve the same number of problems. Older adults showed less cognitive flexibility compared with younger adults in all conditions, a finding that was partially exaggerated in the positive feedback condition. There were no age-group differences in short-term memory; however, for older adults greater perceived resources and positive affect were associated with better memory performance. In sum, older and younger adults were both affected by acute stress, and older adults were not more (or less) vulnerable to the effects of stress on cognition, though they did show stronger associations between self-reported affective states and memory performance. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Effects of cigarette tax on cigarette consumption and the Chinese economy.
To analyse a policy dilemma in China on public health versus the tobacco economy through additional cigarette tax. Using published statistics from 1980 through 1997 to estimate the impact of tobacco production and consumption on government revenue and the entire economy. These estimates relied on the results of estimated price elasticities of the demand for cigarettes in China. Given the estimated price elasticities (-0.54), by introducing an additional 10% increase in cigarette tax per pack (from the current 40% to 50% tax rate), the central government tax revenue would twice exceed total losses in industry revenue, tobacco farmers' income, and local tax revenue. In addition, between 1.44 and 2.16 million lives would be saved by this tax increase. Additional taxation on cigarettes in China would be a desirable public policy for the Chinese government to consider.


Estrogen effects on triglyceride metabolism in analbuminemic rats.
Triglyceride (TG) levels are normally lower in female rats, while the opposite is the case in the Nagase analbuminemic rats (NAR). Increased TG levels in normal males are caused by a testosterone-mediated decrease in postheparin (PH) lipoprotein lipase (LpL). Castration of males reduces TG, while castration of females is without effect. TG levels are reduced by castration of the female NAR, suggesting that estrogen rather than testosterone causes hypertriglyceridemia in this strain. The mechanism for this increase is unknown. We measured secretion of very-low density lipoprotein (VLDL) TG using Triton WR 1339 clearance as the disappearance from blood of 3H-trioleate and 14C-cholesterol-labeled chylomicrons (CM), and the activity of the PH lipases: LpL and hepatic lipase (HL). All were determined in Sprague-Dawley (SD) and NAR female, male, and ovariectomized (OVX) rats. TG levels were significantly greater in female NAR in comparison to all other groups. Ovariectomy of NAR significantly ameliorated hypertriglyceridemia. VLDL TG secretion was significantly greater in intact female NAR compared with all other groups. There were no other differences in VLDL TG secretion among the other groups. The clearance of CM was greatest in female SD rats, and OVX had no effect. NAR cleared CM less well than did SD rats (P < 0.001), but among NAR, clearance was greatest in OVX NAR and male NAR (P < 0. 002). Both PH LpL activity and HL activity were lowest in female NAR (P < 0.05). Ovariectomy partially corrected the defect in HL (P < 0. 05). TG levels in female NAR are in part a result of increased VLDL-TG secretion, an effect mediated by estrogen. The presence of an estrogen-mediated catabolic defect that was alleviated by OVX was also observed. This catabolic defect is likely a result of an estrogen-mediated decrease both in LpL and HL expressed only in the presence of analbuminemia.