Targeted delivery of small interfering RNA to angiogenic endothelial cells with liposome-polycation-DNA particles.
Angiogenesis is an attractive target for cancer therapy, due to its central position in tumor growth and development. Vascular Endothelial Growth Factor (VEGF) and its receptors (VEGFRs) play a key role in the angiogenic process. A promising strategy for targeting VEGF-mediated angiogenesis is RNA interference (RNAi) using short interfering RNA (siRNA). However, for efficacious RNAi a well-designed siRNA delivery system is crucial. Liposome-Polycation-DNA (LPD) particles form a promising system for siRNA delivery to tumors. In order to target angiogenic endothelial cells, LPD particles may be modified with a targeting ligand, such as a cyclic Arg-Gly-Asp (RGD) peptide that specifically binds to integrins expressed on tumor-associated endothelial cells. In the current study, RGD-targeted PEGylated LPD particles containing VEGFR-2 siRNA were prepared and optimized with respect to their size and charge by varying protamine content, carrier DNA content for stronger complexation, and PEGylation density. The size of the optimized particles was around 200 nm and the ζ-potential was approximately +20 mV. The uptake and silencing efficacy of the RGD-targeted PEGylated LPD particles were evaluated in H5V cells (murine endothelial cells) and Human Umbilical Vein Endothelial cells (HUVECs). When compared to non-targeted LPD particles, enhanced uptake and silencing of VEGFR-2 expression was observed for RGD-targeted PEGylated LPD particles. In conclusion, the RGD-targeted PEGylated LPD particles containing VEGFR-2 siRNA presented here may be a promising approach for targeting VEGF-mediated angiogenesis in cancer therapy.


Theoretical analysis of 'addressed' chemical modification of DNA.
Chemical "addressed" modification of DNA involves treatment of single-stranded DNA with oligonucleotides complementary to certain target sequences in this DNA and bearing a groupings reactive towards DNA bases. The binding of oligonucleotides can occur both at completely (specific) and incompletely (nonspecific) complementary sites. We analyse the modification of a fragment that is flanked by two target sequences complementary to a given oligonucleotide address, contains no more such targets and has some randomly distributed sites for nonspecific binding. Conditions for the maximum ratio between specific and non-specific modification are determined. We find the probability of both target termini being specifically modified without any non-specific modification occurring within the fragment up to a given moment in time. Quantitative analysis is based on the use of known features of the specific and non-specific binding of an oligonucleotide to DNA sites. This analysis shows the possibility of specific cutting of DNA based on addressed modification.


Sexual behaviour in context: a global perspective.
Research aimed at investigating sexual behaviour and assessing interventions to improve sexual health has increased in recent decades. The resulting data, despite regional differences in quantity and quality, provide a historically unique opportunity to describe patterns of sexual behaviour and their implications for attempts to protect sexual health at the beginning of the 21st century. In this paper we present original analyses of sexual behaviour data from 59 countries for which they were available. The data show substantial diversity in sexual behaviour by region and sex. No universal trend towards earlier sexual intercourse has occurred, but the shift towards later marriage in most countries has led to an increase in premarital sex, the prevalence of which is generally higher in developed countries than in developing countries, and is higher in men than in women. Monogamy is the dominant pattern everywhere, but having had two or more sexual partners in the past year is more common in men than in women, and reported rates are higher in industrialised than in non-industrialised countries. Condom use has increased in prevalence almost everywhere, but rates remain low in many developing countries. The huge regional variation indicates mainly social and economic determinants of sexual behaviour, which have implications for intervention. Although individual behaviour change is central to improving sexual health, efforts are also needed to address the broader determinants of sexual behaviour, particularly those that relate to the social context. The evidence from behavioural interventions is that no general approach to sexual-health promotion will work everywhere and no single-component intervention will work anywhere. Comprehensive behavioural interventions are needed that take account of the social context in mounting individual-level programmes, attempt to modify social norms to support uptake and maintenance of behaviour change, and tackle the structural factors that contribute to risky sexual behaviour.


Visualization of podocyte substructure with structured illumination microscopy (SIM): a new approach to nephrotic disease.
A detailed microscopic analysis of renal podocyte substructure is essential to understand and diagnose nephrotic kidney disease. Currently only time consuming electron microscopy (EM) can resolve this substructure. We used structured illumination microscopy (SIM) to examine frozen sections of renal biopsies stained with an immunofluorescence marker for podocin, a protein localized to the perimeter of the podocyte foot processes and compared them with EM in both normal and nephrotic disease biopsies. SIM images of normal glomeruli revealed curvilinear patterns of podocin densely covering capillary walls similar to podocyte foot processes seen by EM. Podocin staining of all nephrotic disease biopsies were significantly different than normal, corresponding to and better visualizing effaced foot processes seen by EM. The findings support the first potential use of SIM in the diagnosis of nephrotic disease.


Toward verified biological models.
The last several decades have witnessed a vast accumulation of biological data and data analysis. Many of these data sets represent only a small fraction of the system's behavior, making the visualization of full system behavior difficult. A more complete understanding of a biological system is gained when different types of data (and/or conclusions drawn from the data) are integrated into a larger-scale representation or model of the system. Ideally, this type of model is consistent with all available data about the system, and it is then used to generate additional hypotheses to be tested. Computer-based methods intended to formulate models that integrate various events and to test the consistency of these models with respect to the laboratory-based observations on which they are based are potentially very useful. In addition, in contrast to informal models, the consistency of such formal computer-based models with laboratory data can be tested rigorously by methods of formal verification. We combined two formal modeling approaches in computer science that were originally developed for non-biological system design. One is the inter-object approach using the language of live sequence charts (LSCs) with the Play-Engine tool, and the other is the intra-object approach using the language of statecharts and Rhapsody as the tool. Integration is carried out using InterPlay, a simulation engine coordinator. Using these tools, we constructed a combined model comprising three modules. One module represents the early lineage of the somatic gonad of C. elegans in LSCs, while a second more detailed module in statecharts represents an interaction between two cells within this lineage that determine their developmental outcome. Using the advantages of the tools, we created a third module representing a set of key experimental data using LSCs. We tested the combined statechart-LSC model by showing that the simulations were consistent with the set of experimental LSCs. This small-scale modular example demonstrates the potential for using similar approaches for verification by exhaustive testing of models by LSCs. It also shows the advantages of these approaches for modeling biology.


The consistency between scientific papers presented at the Orthopaedic Trauma Association and their subsequent full-text publication.
To determine the consistency of conclusions/statements made in podium presentations at the annual meeting of the Orthopaedic Trauma Association (OTA) with those in subsequent full-text publications. Also, to evaluate the nature and consistency of study design, methods, sample sizes, results and assign a corresponding level of evidence. Abstracts of the scientific programs of the OTA from 1994 to 1997 (N = 254) were queried by using the PubMed database to identify those studies resulting in a peer-reviewed, full-text publication. Of the 169 articles retrieved, 137 studies were the basis of our study after the exclusion criteria were applied: non-English language, basic science studies, anatomic dissection studies, and articles published in non-peer-reviewed journals. Information was abstracted onto a data form: first from the abstract published in the final meeting program, and then from the published journal article. Information was recorded regarding study issues, including the study design, primary objective, sample size, and statistical methods. We provided descriptive statistics about the frequency of consistent results between abstracts and full-text publications. The results were recorded as percentages and a 95% confidence interval was applied to each value. Study results were recorded for the abstract and full-text publication comparing results and the overall conclusion. A level of scientific-based evidence was assigned to each full-text publication. The final conclusion of the study remained the same 93.4% of the time. The method of study was an observational case series 52% of the time and a statement regarding the rate of patient follow-up was reported 42% of the time. Of the studies published, 18.2% consisted of a sample size smaller than the previously presented abstract. When the published papers had their level of evidence graded, 11% were level I, 16% level II, 17% level III, and 56% level IV. Authors conclusions were consistent with those in full-text publications. Most studies were observational, less than half reported on the rate of patient follow-up. Many abstracts followed by publication had a smaller sample size in the published paper. Half of all studies were graded level IV evidence.


Work capacity assessments and efforts to achieve a job match for claimants in a social security setting: an international inventory.
Many high-income countries are witnessing a shift of focus on eligibility for disability benefits towards promotion of work reintegration. However, little is known about how countries assess work capacity, and how a job match is then obtained. The current study aims to compare work capacity assessments and available efforts to achieve a job match in eight high-income OECD countries. A survey was conducted among key stakeholders concerning organization of work capacity assessments in social security settings, and efforts made to obtain a job, across eight OECD countries: Australia, Denmark, Finland, Iceland, Sweden, the Netherlands, the United Kingdom and the United States. In most countries, work capacity is assessed at several time points, with variations in moments and in information used for job matching. In countries obtaining information on personal and work levels, the search to find a job match usually begins with the persons who have disabilities. Although a shift towards a holistic focus in work capacity assessment has been recognized, medical factors still prevail. Limited emphasis is placed on the implications of functional limitations for the possibilities of work. A holistic approach to assessment needs to be coupled with holistic support measures through provision of coordinated and high quality job matching services.IMPLICATIONS FOR REHABILITATIONBesides determining eligibility for benefits the outcome of the work capacity assessment can also be used for other purposes such as reintegration and should not result in a static description of work capacity but also deliver insight in support needs.Involving multiple institutions and disciplines in work capacity assessments may result in a broader overview of the claimants' capacities, however this places high demands on cooperation and data sharing of all those involved.Incorporating the claimants own perspective on work capacity and possibilities to work might improve the legitimacy of the process and reasonably also the achievement of a good and sustainable job match. This can be achieved by using self reported questionnaires and interviews in the assessments.


Locating systematic reviews of test accuracy studies: how five specialist review databases measure up.
The aim of this study was to examine location of systematic reviews of test accuracy in five specialist review databases: York CRD's DARE and HTA databases, Medion (University of Maastricht), C-EBLM (International Federation of Clinical Chemistry), and the ARIF in-house database (University of Birmingham). Searches were limited to the period 1996-2006. Test accuracy reviews were located using in-house diagnostic search filters and with help from database producers where databases were not confined to test accuracy reviews. References were coded according to disease area, review purpose, and test application. Ease of use, volume, overlap, and content of databases was noted. A large degree of overlap existed between databases. Medion contained the largest number (n = 672) and the largest number of unique (n = 328) test accuracy references. A combination of three databases identified only 76% of test reviews. All databases were rated as easy to search but varied with respect to timeliness and compatibility with reference management software. Most reviews evaluated test accuracy (85%) but the HTA database had a larger proportion of cost-effectiveness and screening reviews and C-EBLM more reviews addressing early test development. Most reviews were conducted in secondary care settings. Specialist review databases offer an essential addition to general bibliographic databases where application of diagnostic method filters can compromise search sensitivity. Important differences exist between databases in terms of ease of use and content. Our findings raise the question whether the current balance of research setting, in particular the predominance of research on tests used in secondary care, matches the needs of decision makers.


The characteristics and motivations of online health information seekers: cross-sectional survey and qualitative interview study.
Most households in the United Kingdom have Internet access, and health-related Internet use is increasing. The National Health Service (NHS) Direct website is the major UK provider of online health information. Our objective was to identify the characteristics and motivations of online health information seekers accessing the NHS Direct website, and to examine the benefits and challenges of the health Internet. We undertook an online questionnaire survey, offered to users of the NHS Direct website. A subsample of survey respondents participated in in-depth, semistructured, qualitative interviews by telephone or instant messaging/email. Questionnaire results were analyzed using chi-square statistics. Thematic coding with constant comparison was used for interview transcript analysis. In total 792 respondents completed some or all of the survey: 71.2% (534/750 with data available) were aged under 45 years, 67.4% (511/758) were female, and 37.7% (286/759) had university-level qualifications. They sought information for themselves (545/781, 69.8%), someone else (172/781, 22.0%), or both (64/781, 8.2%). Women were more likely than men to seek help for someone else or both themselves and someone else (168/509 vs 61/242, χ(2) (2) = 6.35, P = .04). Prior consultation with a health professional was reported by 44.9% (346/770), although this was less common in younger age groups (<36 years) (χ(2) (1) = 24.22, P < .001). Participants aged 16 to 75 years (n = 26, 20 female, 6 male) were recruited for interview by telephone (n = 23) and instant messaging/email (n = 3). Four major interview themes were identified: motivations for seeking help online; benefits of seeking help in this way and some of the challenges faced; strategies employed in navigating online health information provision and determining what information to use and to trust; and specific comments regarding the NHS Direct website service. Within the motivation category, four concepts emerged: the desire for reassurance; the desire for a second opinion to challenge other information; the desire for greater understanding to supplement other information; and perceived external barriers to accessing information through traditional sources. The benefits clustered around three theme areas: convenience, coverage, and anonymity. Various challenges were discussed but no prominent theme emerged. Navigating online health information and determining what to trust was regarded as a "common sense" activity, and brand recognition was important. Specific comments about NHS Direct included the perception that the online service was integrated with traditional service provision. This study supports a model of evolutionary rather than revolutionary change in online health information use. Given increasing resource constraints, the health care community needs to seek ways of promoting efficient and appropriate health service use, and should aim to harness the potential benefits of the Internet, informed by an understanding of how and why people go online for health.


Dehydroepiandrosterone sulfate and anovulation increase serum inhibin and affect follicular function during administration of gonadotropins.
To test the hypothesis that testosterone (T) derived from circulating dehydroepiandrosterone sulfate (DS) or produced in excess within the anovulatory ovary is a follicular regulator capable of stimulating inhibin secretion. DS and inhibin were determined by RIA in residual serum samples from in vitro fertilization cycles and analyzed along with other parameters using analysis of variance and stepwise multiple linear regression after log transformation of the RIA data. It was predicted that the serum concentration of inhibin would be higher in women with anovulation than in normal subjects and that the serum inhibin concentration would have a positive regression coefficient on the serum DS concentration. The serum concentrations of inhibin (P < 0.01) and estradiol (P < 0.001) on the day of hCG treatment were higher in women with anovulation than in normal subjects, as was the number of oocytes retrieved (P < 0.001). The FSH and LH doses (both P < 0.005) and age (P < 0.001) were significantly lower, and the average duration of gonadotropin therapy was 1 day shorter (P < 0.001) in anovulatory patients. There was no significant difference in serum DS (P > 0.2). Regression analysis showed that serum inhibin was positively related to the number of oocytes (P < 0.001) and serum DS (P = 0.05), with an increase in anovulatory subjects (P < 0.025). The duration of treatment with gonadotropins was related positively to the patient's age (P < 0.001) and negatively to serum DS (P < 0.025), with a decrease in anovulatory subjects (P < 0.025). The number of oocytes obtained was negatively related to age (P < 0.001) and positively to serum DS (P < 0.005). These data are consistent with a stimulatory effect of follicular T derived from either circulating DS or the anovulatory ovary, which affects the secretion of inhibin, the duration of gonadotropin treatment, and the number of oocytes retrieved.


The myxoma virus-soluble interferon-gamma receptor homolog, M-T7, inhibits interferon-gamma in a species-specific manner.
The myxoma virus M-T7 protein contains significant sequence similarity to the ligand binding domain of the mammalian interferon-gamma receptors, and functions as a soluble homolog which can bind and inhibit the biological activities of rabbit interferon-gamma (Upton, C., Mossman, K., and McFadden, G. (1992) Science 258:1369-1372). M-T7, the most abundantly secreted protein from myxoma virus-infected cells, was shown to be expressed in significant biological amounts as a typical poxvirus early gene product, efficiently secreted at early times of infection to levels that exceed 5 x 10(7) molecules/cell, and function as a stable inhibitory protein in infected cell supernatants until late times of infection. M-T7 was specific in binding and inhibiting rabbit interferon-gamma, and did not bind either human or murine interferon-gamma. Scatchard analysis of rabbit interferon-gamma binding curves yielded a single high affinity binding site on M-T7, with a Kd of 1.2 x 10(-9) M, which is comparable to the affinity between soluble forms of cellular interferon-gamma receptors and their cognate ligands. In comparison, rabbit interferon-gamma was shown to bind its cellular receptor with a Kd of 5.9 x 10(-10) M, again comparable to the affinity of membrane bound forms of other mammalian interferon-gamma receptors for interferon-gamma. Thus, the myxoma virus M-T7 protein is a functional soluble interferon-gamma receptor homolog which binds and inhibits interferon-gamma with high affinity in a species-specific manner.


Mammographic breast density, dense area, and breast area differences by phase in the menstrual cycle.
Mammographic breast density may be greater in the luteal phase (days 15-30) than the follicular phase (days 1-14) of the menstrual cycle; this may have implications for when mammography screening should occur. Examine whether percent breast density, breast area, or dense area differ by menstrual phase. We identified 204 premenopausal women with regular periods who were <55 years (mean = 45.0 years) and had two screening mammograms within 9 to 18 months, with one screening between days 9 and 14, and one screening between days 22 and 35 of the menstrual cycle. We measured percent breast density, breast area, and dense area using the Cumulus software. We used linear regression to test for differences in breast density, breast area, and dense area from follicular to luteal phase, adjusting for change in weight and time between exams. The mean (SD) percent breast density was 35.8% (21.3) in the follicular phase and 36.7% (21.3) in the luteal phase. Multivariable analyses showed small but not statistically significant increases in percent density [1.1%; 95% confidence interval (95% CI), -0.2% to 2.3%] and breast area (16.7 cm(2); 95% CI, -2.8 to 36.2) and a statistically significant increase in dense area (13.1 cm(2); 95% CI, 0.1-26.1) in the luteal compared with the follicular phase. Breast density, breast area, and dense area have small, but probably not clinically meaningful, increases in the luteal phase of the menstrual cycle. However, there are other factors that may differ by menstrual cycle phase that we were unable to assess (e.g., breast compression), which may ultimately influence mammographic sensitivity by menstrual cycle phase.


Analysis of starch distribution in the paper cross-section by Raman microscopy.
A new Raman microscopy approach was developed to analyze the starch distribution of paper cross-sections in a faster and more specific way than is possible with the currently used iodine-staining method. Raman images were recorded and analyzed from cross-sections of cellulose hand sheets surface-sized with 1% or 2% starch solutions and with different film thicknesses. In addition, Raman imaging analysis of the starch distribution was performed on two industrial papers, an abrasive base paper and a surface-sized recycling paper. The visualization and the quantitative analysis of the starch distribution were performed by using the intensity changes of the Raman starch band at 855 cm(-1) and by principal component analysis. Distribution curves were calculated from the intensity data and compared for the samples with different starch concentrations and with results obtained from iodine-stained cross-sections of the same samples. The results of this study demonstrate the great potential and the new possibilities of Raman microscopy for studying the z-distribution of chemical components and additives in paper.


A catechol derivative (4-methylcatechol) accelerates the recovery from experimental acrylamide-induced neuropathy.
Acrylamide (ACR) monomer produces neuropathy of the dying-back type and 4-methylcatechol (4-MC) is a potent stimulator of endogenous nerve growth factor synthesis. In the present study, we investigated the efficacy of 4-MC in promoting recovery from experimental ACR neuropathy in rats. Twenty-two Sprague-Dawley rats were made neuropathic by ACR injections. They showed hindlimb paralysis, increment of landing foot spread distance and a statistically significant reduction in motor nerve conduction velocity. After the ACR neuropathy had been established, 12 of the rats were administered 4-MC for 2 weeks, and the other 10 were injected with phosphate-buffered saline alone. 4-MC-administered ACR neuropathy rats showed improvement, i.e., a decrease in landing foot spread distance, increase in motor nerve conduction velocity and increase in nerve growth factor content in the sciatic nerves in comparison with the corresponding values for ACR neuropathy rats given phosphate-buffered saline alone. A decreased number of large myelinated fiber with a reciprocal increase in small myelinated fiber number also was seen in the ACR neuropathy rats; however, this change was ameliorated in part by the administration of 4-MC. Therefore, these findings suggest that 4-MC can accelerate the recovery process clinically, electrophysiologically, biochemically and neuropathologically.


Impairment of DNA Methylation Maintenance Is the Main Cause of Global Demethylation in Naive Embryonic Stem Cells.
Global demethylation is part of a conserved program of epigenetic reprogramming to naive pluripotency. The transition from primed hypermethylated embryonic stem cells (ESCs) to naive hypomethylated ones (serum-to-2i) is a valuable model system for epigenetic reprogramming. We present a mathematical model, which accurately predicts global DNA demethylation kinetics. Experimentally, we show that the main drivers of global demethylation are neither active mechanisms (Aicda, Tdg, and Tet1-3) nor the reduction of de novo methylation. UHRF1 protein, the essential targeting factor for DNMT1, is reduced upon transition to 2i, and so is recruitment of the maintenance methylation machinery to replication foci. Concurrently, there is global loss of H3K9me2, which is needed for chromatin binding of UHRF1. These mechanisms synergistically enforce global DNA hypomethylation in a replication-coupled fashion. Our observations establish the molecular mechanism for global demethylation in naive ESCs, which has key parallels with those operating in primordial germ cells and early embryos.


Myasthenia gravis: diagnostic and management dilemmas.
This review focuses on the diagnostic tests that may help to confirm myasthenia in patients without acetylcholine receptor antibodies, including the newly discovered anti-muscle-specific receptor tyrosine kinase antibody and other investigations. We discuss management dilemmas, particularly those that revolve around treatments whose efficacy is questionable or unproved, such as thymectomy. We review the important issue of how treatments are assessed, and examine future treatment trial designs.


Noise induced hearing loss risk assessment in truck drivers.
Hearing sense is one of the key elements which may have impact on the driver's task quality. This cross-sectional study investigates the hearing status of 500 truck drivers by pure tone audiometry (AC) in one of the cities in Fars province, Iran. Hearing threshold levels of the subjects were measured in frequencies of 500Hz-8000Hz. Screening and determination of permanent threshold shift (PTS) was the first aim of this study. Hence tests were done at least 16 hours after any exposure to noticeable sound. The effect of age as a confounding factor was considered using ISO equation and subtracted from whole hearing threshold. The threshold of 25 dB HL and above was considered abnormal but the calculation of hearing was also carried out using 0 dB HL as reference. Subjects were categorized into two groups on the basis of working experience and the hearing threshold of 25 dB was considered a boundary of normal hearing sense. The results of Pearson Chi-Square test showed that working experience as an independent variable has significant contributing effect on hearing thresholds of truck drivers in frequencies of 500, 1000, 2000 and 4000 Hz (p greater than 0.05). Also, it was shown that currently nine and 12.6 % of truck drivers suffer from impaired hearing sense in left and right respectively (hearing threshold level greater than 25 dB) in mid frequencies (500, 1000, 2000 Hz) and 45% in high frequencies of both ears (4000 and 8000 Hz). The results indicated that hearing damage of professional drivers was expected to occur sooner at 4000 and 8000 Hz than lower frequencies. Finally it was deduced that the occupational conditions of truck drivers may have bilateral, symmetrical harmful effect on hearing threshold sense in all frequencies mainly in frequency of 4000 Hz, so health surveillance programs such as education and periodic medical examinations are emphasized for pre-diagnosing and prevention of any possible impairment and an urgent need to take up some interventions such as better maintenance of roads, automobile industry efforts to reduce the noise level emission of vehicles and reducing number of working hours per day of drivers are highlighted to improve the harmful working conditions of truck drivers.


A single-molecule digital enzyme assay using alkaline phosphatase with a cumarin-based fluorogenic substrate.
Digitalization of fluorogenic enzymatic assays through the use of femtoliter chamber array technology is an emerging approach to realizing highly quantitative bioassays with single-molecule sensitivity. However, only a few digital fluorogenic enzyme assays have been reported, and the variations of the digital enzyme assays are basically limited to fluorescein- and resorufin-based fluorogenic assays. This limitation hampers the realization of a multiplex digital enzyme assay such as a digital enzyme-linked immunosorbent assay (ELISA). In this study, after optimization of buffer conditions, we achieved a single-molecule digital enzyme alkaline phosphatase (ALP) assay with a cumarin-based fluorogenic substrate, 4-methylunbelliferyl phosphate (4-MUP). When ALP molecules were encapsulated in a 44-femtoliter chamber array at a low ratio of less than 1 molecule per chamber, each chamber showed a discrete fluorescence signal in an all-or-none manner, allowing the digital counting of the number of active enzyme molecules. The fraction of fluorescent chambers linearly decreased with the enzyme concentration, obeying the Poisson distribution as expected. We also demonstrated a dual-color digital enzyme assay with a ALP/4-MUP and β-galactosidase (β-gal)/resorufin-β-d-galactopyranoside combination. The activities of single ALP and β-gal molecules were clearly detected simultaneously. The method developed in this study will enable us to carry out a parallelized, multiplex digital ELISA.


NAADP, a new intracellular messenger that mobilizes Ca2+ from acidic stores.
NAADP (nicotinic acid-adenine dinucleotide phosphate) is a recently described Ca2+-mobilizing molecule. First characterized in the sea urchin egg, it has been shown to mobilize Ca2+ from intracellular stores in a wide range of cells from different organisms. It is a remarkably potent molecule, and recent reports show that its cellular levels change in response to a variety of agonists, confirming its role as a Ca2+-mobilizing messenger. In many cases, NAADP appears to interact with other Ca2+-mobilizing messengers such as IP3 (inositol 1,4,5-trisphosphate) and cADP-ribose in shaping cytosolic Ca2+ signals. What is not clear is the molecular nature of the NAADP-sensitive Ca2+ release mechanism and its subcellular localization. This review focuses on the recent progress made in sea urchin eggs, which indicates that NAADP activates a novel Ca2+ release channel distinct from the relatively well-characterized IP3 and ryanodine receptors. Furthermore, in the sea urchin egg, the NAADP-sensitive store appears to be separate from the endoplasmic reticulum and is most likely an acidic store. These findings have also been reinforced by similar findings in mammalian cells, and a unified model for NAADP-induced Ca2+ signalling is presented.


The impact of implementation of the Canadian regulatory requirements on the quality of natural health products: the glucosamine case.
We investigated whether the recent implementation of the regulatory requirements for the entry to the Canadian market of natural products has resulted in improved quality of the available glucosamine products. Eleven available products, of which 8 had been tested in 2002 (7 had contained substantially lower than the label claim of the active ingredient), and a European pharmaceutical grade tablet were assayed for their glucosamine content. The potassium and sodium contents of the products were also tested. Nine of the 11 Canadian products and the European tablet had more than 91% of the label claim of the active ingredient, hence, met the criterion. Two products contained 71 and 78% label claim. The electrolyte contents were very variable but constituted only a small fraction of the daily requirements. Most tested glucosamine products passed the Health Canada requirements. This improvement is likely due to the publicity regarding the low quality of the products in the past and also a result, at least in part, of the introduction of the new regulatory requirements. The sub-standard quality of a few tested products is still of concern.


Recognition and Management of Hypertension in Older Persons: Focus on African Americans.
Hypertension is the most commonly diagnosed condition in persons aged 60 and older and is the single most important risk factor for cardiovascular disease (ischemic heart disease, heart failure, and stroke), kidney disease, and dementia. More than half of individuals with hypertension in the United States are aged 60 and older. Hypertension disproportionately affects African Americans, with all age groups, including elderly adults, having a higher burden of hypertension-related complications than other U.S. Multiple clinical trials have demonstrated the beneficial effects of blood pressure (BP) reduction on cardiovascular morbidity and mortality, with most of the evidence in individuals aged 60 and older. Several guidelines have recently been published on the specific management of hypertension in individuals aged 60 and older, including in high-risk groups such as African Americans. Most recommend careful evaluation, thiazide diuretics and calcium-channel blockers for initial drug therapy in most African Americans, and angiotensin-converting enzyme inhibitors and angiotensin receptor blockers in those with chronic kidney disease or heart failure. Among the areas of controversy is the recommended target BP in African Americans aged 60 and older. A recent U.S. guideline recommended raising the systolic BP target from less than 140 mmHg to less than 150 mmHg in this population. This article will review the evidence and current guideline recommendations for hypertension treatment in older African Americans, including the rationale for continuing to recommend a SBP target of less than 140 mmHg in this population.


Hemodynamic parameters in blood vessels in choroidal melanoma xenografts and rat choroid.
Choroidal melanoma is the most common primary ocular cancer among the adult population. To avoid enucleation, there has been a concerted effort to develop therapies that spare the affected eye and the patient's vision. Blood flow helps shape the tumor's microenvironment, plays a key role in the tumor's response to many different types of therapy, and is necessary for delivery of chemotherapeutic agents. To rationally design new therapies and optimize existing treatments, it is essential to learn as much as possible about blood flow and the microcirculation in this tumor. In recent years, much has been discovered about the anatomy of the microvasculature and the dynamics of overall blood flow in choroidal melanoma, but little is known about the factors that determine microvascular blood flow. In this study hemodynamic parameters in individual microvessels of a human choroidal melanoma xenograft were compared with those same parameters in a normal rat choroid. Nude, athymic WAG/RijHs-rnu rats were used in this study. The human choroidal melanoma cell line OCM-1 was used to grow solid tumors subcutaneously in the flanks of donor rats. Small pieces of these tumors were then implanted into the choroidal space of recipient rats. After 6 to 8 weeks, the rats were anesthetized with a subcutaneous injection of urethane, and the sclera was exposed. Rhodamine-labeled liposomes and red blood cells (RBCs) labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) were injected intravenously. Epifluorescent, intravital microscopy was used to visualize the flow of fluorescent RBCs through individual vessels in the choroid or tumor. Flow through multiple vessels was recorded on videotape for later analysis. From the recordings, RBC flux, RBC velocity (V(c)), and microvascular hematocrit (HCT(m)) were determined. Similar experiments were performed in rats with no tumor growth, and these same parameters were calculated in normal choroidal vessels. RBC flow was characterized in 55 vessels in six OCM-1 tumors and in 153 choroidal vessels in five non-tumor-bearing rats. RBC flux was higher in larger tumor vessels (>30 micro m in diameter) compared with similarly sized choroidal vessels. There were no differences in the velocities of RBCs through the two types of vessels. HCT(m) was significantly higher in medium-sized (>20 micro m in diameter) and larger tumor vessels compared with normal choroidal vessels. These experiments demonstrate differences between hemodynamic parameters in normal choroidal microvessels and microvessels in choroidal melanoma in this animal model. Because HCT(m) is a key determinant of apparent viscosity, abnormally high HCT(m) in the tumor vessels would increase vascular resistance and decrease flow. This could have a negative impact on the tumor oxygen levels and on the ability to deliver drugs effectively. On the contrary, higher local HCT(m) has also been shown to increase oxygen delivery. The impact and interplay of these two effects on tumor oxygenation remain to be determined.


Growth of healthy infants and the timing, type, and frequency of complementary foods.
Growth patterns of exclusively and predominantly breast-fed infants differ from those of non-breast-fed infants, but less is known about associations among growth patterns and different durations of exclusive breast-feeding and the types and frequency of complementary foods. We examined these associations, particularly between 4 and 6 mo of age, using data from a unique longitudinal 7-country study. Data from the World Health Organization Multinational Study of Breast-feeding and Lactational Amenorrhea on infants living in generally favorable environments were used. Multilevel analyses described growth and the relation between growth and variables related to feeding. Small differences in growth that were statistically significant but probably not biologically important were noted among infants in whom complementary foods were introduced at different times. Weight gain was more sensitive to feeding frequencies than were gains in length, but the cumulative 10-wk differences were small. The most extreme differences were equivalent to approximately 10 centiles of the weight and height distributions at 6 mo of age. These results do not provide compelling evidence of benefit or risk related to growth and the timing of introduction of complementary foods at any specific time between 4 and 6 mo of age. Thus, postnatal growth appears to not be sensitive to the differential timing of introduction of complementary foods nor to differential types and frequencies of complementary foods in healthy infants living in environments without major economic constraints and low rates of illness. These results, however, may not indicate growth differences in populations living in poor environments.


Auxin regulation of the gibberellin pathway in pea.
The auxin indole-3-acetic acid (IAA) has been shown to promote the biosynthesis of the active gibberellin (GA(1)) in shoots of pea (Pisum sativum). We used northern analysis to investigate the timing of IAA-induced changes in transcript levels of PsGA3ox1 (Mendel's LE), PsGA2ox1, PsGA2ox2, and PsGA20ox1, key genes for the later stages of GA(1) biosynthesis and metabolism in pea. Rapid (2-4 h) changes occurred in the transcript levels of PsGA3ox1, PsGA2ox1, and PsGA2ox2 after treatment with IAA. [(14)C]GA(1) metabolism studies in decapitated shoots indicated that IAA inhibits GA(1) deactivation, consistent with the suppression of PsGA2ox1 (SLN) transcript levels by IAA. Studies with the sln mutant also indicated that PsGA2ox1 activity is involved in GA(1) deactivation in decapitated shoots. Culture of excised internode tissue in the presence of auxin clearly demonstrated that internode tissue is a site of GA(1) biosynthesis per se. Excised internode tissue cultured in the presence/absence of cycloheximide showed that de novo protein synthesis is required for IAA-induced increases in PsGA3ox1 transcript levels. Auxin dose response studies indicated that IAA concentration is a critical determinant of GA(1) biosynthesis over 1 to 2 orders of magnitude, and a range of auxins was shown to affect the GA pathway.


Evidence for subjective values guiding posture and movement coordination in a free-endpoint whole-body reaching task.
When moving, humans must overcome intrinsic (body centered) and extrinsic (target-related) redundancy, requiring decisions when selecting one motor solution among several potential ones. During classical reaching studies the position of a salient target determines where the participant should reach, constraining the associated motor decisions. We aimed at investigating implicit variables guiding action selection when faced with the complexity of human-environment interaction. Subjects had to perform whole body reaching movements towards a uniform surface. We observed little variation in the self-chosen motor strategy across repeated trials while movements were variable across subjects being on a continuum from a pure 'knee flexion' associated with a downward center of mass (CoM) displacement to an 'ankle dorsi-flexion' associated with an upward CoM displacement. Two optimality criteria replicated these two strategies: a mix between mechanical energy expenditure and joint smoothness and a minimization of the amount of torques. Our results illustrate the presence of idiosyncratic values guiding posture and movement coordination that can be combined in a flexible manner as a function of context and subject. A first value accounts for the reach efficiency of the movement at the price of selecting possibly unstable postures. The other predicts stable dynamic equilibrium but requires larger energy expenditure and jerk.


Overexpression of Bcl-2 inhibits alveolar cell apoptosis during involution and accelerates c-myc-induced tumorigenesis of the mammary gland in transgenic mice.
Expression of the apoptosis-inhibitory protein Bcl-2 has frequently been detected in human cancer including mammary carcinoma. The functional significance of its expression has been well established in experimental tumors of the lymphoid system, however, remains to be elucidated for epithelial tumors. In order to assess the role of Bcl-2 in mammary tumorigenesis we have generated WAP-bcl-2 transgenic mice. The strong overexpression of Bcl-2 in lactating mammary glands was preserved during early postlactational involution and apoptosis of alveolar epithelial cells was prevented without influencing the dedifferentiation of the milk-producing epithelium. Although Bcl-2 overexpression was not sufficient to induce spontaneous tumors it, however, led to an accelerated development of MMTV myc transgene-induced mammary tumors. In the mammary glands of MMTV myc transgenic mice, a high proportion of apoptotic cells was detected which was significantly reduced in the mammary glands of WAP-bcl-2/ MMTV myc double transgenic mice. Taken together, these results suggest that Bcl-2 contributes to mammary tumorigenesis by inhibiting apoptosis.


Direct quantification of inorganic polyphosphate in microbial cells using 4'-6-diamidino-2-phenylindole (DAPI).
Inorganic polyphosphate (polyP) is increasingly being recognized as an important phosphorus sink within the environment, playing a central role in phosphorus exchange and phosphogenesis. Yet despite the significant advances made in polyP research there is a lack of rapid and efficient analytical approaches for the quantification of polyP accumulation in microbial cultures and environmental samples. A major drawback is the need to extract polyP from cells prior to analysis. Due to extraction inefficiencies this can lead to an underestimation of both intracellular polyP levels and its environmental pool size: we observed 23-58% loss of polyP using standard solutions and current protocols. Here we report a direct fluorescence based DAPI assay system which removes the requirement for prior polyP extraction before quantification. This increased the efficiency of polyP detection by 28-55% in microbial cultures suggesting quantitative measurement of the intracellular polyP pool. It provides a direct polyP assay which combines quantification capability with technical simplicity. This is an important step forward in our ability to explore the role of polyP in cellular biology and biogeochemical nutrient cycling.


Mitoriboscins: Mitochondrial-based therapeutics targeting cancer stem cells (CSCs), bacteria and pathogenic yeast.
The "endo-symbiotic theory of mitochondrial evolution" states that mitochondrial organelles evolved from engulfed aerobic bacteria, after millions of years of symbiosis and adaptation. Here, we have exploited this premise to design new antibiotics and novel anti-cancer therapies, using a convergent approach. First, virtual high-throughput screening (vHTS) and computational chemistry were used to identify novel compounds binding to the 3D structure of the mammalian mitochondrial ribosome. The resulting library of ∼880 compounds was then subjected to phenotypic drug screening on human cancer cells, to identify which compounds functionally induce ATP-depletion, which is characteristic of mitochondrial inhibition. Notably, the top ten "hit" compounds define four new classes of mitochondrial inhibitors. Next, we further validated that these novel mitochondrial inhibitors metabolically target mitochondrial respiration in cancer cells and effectively inhibit the propagation of cancer stem-like cells in vitro. Finally, we show that these mitochondrial inhibitors possess broad-spectrum antibiotic activity, preventing the growth of both gram-positive and gram-negative bacteria, as well as C. albicans - a pathogenic yeast. Remarkably, these novel antibiotics also were effective against methicillin-resistant Staphylococcus aureus (MRSA). Thus, this simple, yet systematic, approach to the discovery of mitochondrial ribosome inhibitors could provide a plethora of anti-microbials and anti-cancer therapies, to target drug-resistance that is characteristic of both i) tumor recurrence and ii) infectious disease. In summary, we have successfully used vHTS combined with phenotypic drug screening of human cancer cells to identify several new classes of broad-spectrum antibiotics that target both bacteria and pathogenic yeast. We propose the new term "mitoriboscins" to describe these novel mitochondrial-related antibiotics. Thus far, we have identified four different classes of mitoriboscins, such as: 1) mitoribocyclines, 2) mitoribomycins, 3) mitoribosporins and 4) mitoribofloxins. However, we broadly define mitoriboscins as any small molecule(s) or peptide(s) that bind to the mitoribosome (large or small subunits) and, as a consequence, inhibit mitochondrial function, i.e., mitoribosome inhibitors.


Microstructured polymer optical fiber for long period gratings fabrication using an ultraviolet laser beam.
We present a microstructured polymer fiber dedicated to long period grating (LPGs) inscription using a focused UV laser beam. The core and the microstructured cladding of the developed fiber are made of pure poly(methyl methacrylate) (PMMA). The external layer of the solid part of the cladding has increased UV absorption due to doping with trans-4-stilbenemethanol, which shows an absorption band at around 310 nm related to trans-cis photoisomerization. We present transmission characteristics of LPGs fabricated in this fiber using the point-by-point inscription technique with a He-Cd laser beam of 325 nm wavelength. We also demonstrate that in the proposed fiber, the fabrication process is shortened six times compared to pure PMMA fibers. Moreover, we report on temperature response and long-term stability of the fabricated gratings.


mRNA decay proteins are targeted to poly(A)+ RNA and dsRNA-containing cytoplasmic foci that resemble P-bodies in Entamoeba histolytica.
In higher eukaryotes, mRNA degradation and RNA-based gene silencing occur in cytoplasmic foci referred to as processing bodies (P-bodies). In protozoan parasites, the presence of P-bodies and their putative role in mRNA decay have yet to be comprehensively addressed. Identification of P-bodies might provide information on how mRNA degradation machineries evolved in lower eukaryotes. Here, we used immunofluorescence and confocal microscopy assays to investigate the cellular localization of mRNA degradation proteins in the human intestinal parasite Entamoeba histolytica and found evidence of the existence of P-bodies. Two mRNA decay factors, namely the EhXRN2 exoribonuclease and the EhDCP2 decapping enzyme, were localized in cytoplasmic foci in a pattern resembling P-body organization. Given that amoebic foci appear to be smaller and less rounded than those described in higher eukaryotes, we have named them "P-body-like structures". These foci contain additional mRNA degradation factors, including the EhCAF1 deadenylase and the EhAGO2-2 protein involved in RNA interference. Biochemical analysis revealed that EhCAF1 co-immunoprecipitated with EhXRN2 but not with EhDCP2 or EhAGO2-2, thus linking deadenylation to 5'-to-3' mRNA decay. The number of EhCAF1-containing foci significantly decreased after inhibition of transcription and translation with actinomycin D and cycloheximide, respectively. Furthermore, results of RNA-FISH assays showed that (i) EhCAF1 colocalized with poly(A)(+) RNA and (ii) during silencing of the Ehpc4 gene by RNA interference, EhAGO2-2 colocalized with small interfering RNAs in cytoplasmic foci. Our observation of decapping, deadenylation and RNA interference proteins within P-body-like foci suggests that these structures have been conserved after originating in the early evolution of eukaryotic lineages. To the best of our knowledge, this is the first study to report on the localization of mRNA decay proteins within P-body-like structures in E. histolytica. Our findings should open up opportunities for deciphering the mechanisms of mRNA degradation and RNA-based gene silencing in this deep-branching eukaryote.


First and second trimester urinary metabolic profiles and fetal growth restriction: an exploratory nested case-control study within the infant development and environment study.
Routine prenatal care fails to identify a large proportion of women at risk of fetal growth restriction (FGR). Metabolomics, the comprehensive analysis of low molecular weight molecules (metabolites) in biological samples, can provide new and earlier biomarkers of prenatal health. Recent research has suggested possible predictive first trimester urine metabolites correlating to fetal growth restriction in the third trimester. Our objective in this current study was to examine urinary metabolic profiles in the first and second trimester of pregnancy in relation to third trimester FGR in a US population from a large, multi-center cohort study of healthy pregnant women. We conducted a nested case-control study within The Infant Development and the Environment Study (TIDES), a population-based multi-center pregnancy cohort study. We identified 53 cases of FGR based on the AUDIPOG [Neonatal growth - AUDIPOG [Internet]. [cited 29 Nov 2016]. Available from: http://www.audipog.net/courbes_morpho.php?langue=en ] formula for birthweight percentile considering maternal height, age, and prenatal weight, as well as infant sex, gestational age, and birth rank. Cases were matched to 106 controls based on study site, maternal age (± 2 years), parity, and infant sex. NMR spectroscopy was used to assess concentrations of four urinary metabolites that have been previously associated with FGR (tyrosine, acetate, formate, and trimethylamine) in first and second trimester urine samples. We fit multivariate conditional logistic regression models to estimate the odds of FGR in relation to urinary concentrations of these individual metabolites in the first and second trimesters. Exploratory analyses of custom binned spectroscopy results were run to consider other potentially related metabolites. We found no significant association between the relative concentrations of each of the four metabolites and odds of FGR. Exploratory analyses did not reveal any significant differences in urinary metabolic profiles. Compared with controls, cases delivered earlier (38.6 vs 39.8, p < 0.001), and had lower birthweights (2527 g vs 3471 g, p < 0.001). Maternal BMI was similar between cases and controls. First and second trimester concentrations of urinary metabolites (acetate, formate, trimethylamine and tyrosine) did not predict FGR. This inconsistency with previous studies highlights the need for more rigorous investigation and data collection in this area before metabolomics can be clinically applied to obstetrics.


Fondaparinux vs enoxaparin for the prevention of venous thromboembolism in major orthopedic surgery: a meta-analysis of 4 randomized double-blind studies.
Orthopedic surgery remains a condition at high risk of venous thromboembolism (VTE). Fondaparinux, the first of a new class of synthetic selective factor Xa inhibitors, may further reduce this risk compared with currently available thromboprophylactic treatments. A meta-analysis of 4 multicenter, randomized, double-blind trials in patients undergoing elective hip replacement, elective major knee surgery, and surgery for hip fracture (N = 7344) was performed to determine whether a subcutaneous 2.5-mg, once-daily regimen of fondaparinux sodium starting 6 hours after surgery was more effective and as safe as approved enoxaparin regimens in preventing VTE. The primary efficacy outcome was VTE up to day 11, defined as deep vein thrombosis detected by mandatory bilateral venography or documented symptomatic deep vein thrombosis or pulmonary embolism. The primary safety outcome was major bleeding. Fondaparinux significantly reduced the incidence of VTE by day 11 (182 [6.8%] of 2682 patients) compared with enoxaparin (371 [13.7%] of 2703 patients), with a common odds reduction of 55.2% (95% confidence interval, 45.8% to 63.1%; P<.001); this beneficial effect was consistent across all types of surgery and all subgroups. Although major bleeding occurred more frequently in the fondaparinux-treated group (P =.008), the incidence of clinically relevant bleeding (leading to death or reoperation or occurring in a critical organ) did not differ between groups. In patients undergoing orthopedic surgery, 2.5 mg of fondaparinux sodium once daily, starting 6 hours postoperatively, showed a major benefit over enoxaparin, achieving an overall risk reduction of VTE greater than 50% without increasing the risk of clinically relevant bleeding.


Early Diagnosis and Treatment of Coronary Heart Disease in Asymptomatic Subjects With Advanced Vascular Atherosclerosis of the Carotid Artery (Type III and IV b Findings Using Ultrasound) and Risk Factors.
A study was conducted as to whether the early diagnosis of coronary heart disease in asymptomatic subjects with advanced atherosclerosis of the carotid artery which additionally shows at least one risk factor is successful using ultrasound technology. Within the scope of an occupational screening program using subjects from diverse employment sectors, people were given the opportunity to determine their risk of heart attack. During the study the total plaque area (TPA), the maximum plaque thickness in the carotid artery and the PROCAM-Scores of 3,748 healthy men and 2,260 healthy women between the ages of 20 and 64 years were determined. During the subsequent follow-up study 94 subjects sickened. An ultrasound examination of the carotid artery of 79 patients revealed a type III or IV b finding. In a pilot study 33 asymptomatic subjects with a type III or IV b finding in the ultrasound examination were assessed using a computed tomography (CT) coronary angiogram. Additional 10 asymptomatic subjects were examined independently to undergo further cardiac examinations. In the final analysis only five patients had entirely smooth coronary arteries, six had coronary sclerosis, eight had a 30% stenosis, one had a 30-50% stenosis and 23 patients had a stenosis ≥ 50%; and in extreme case, a left main coronary artery stenosis with three-vessel disease. Asymptomatic subjects with advanced atherosclerosis of the carotid artery (type III and type IV b findings) had a high risk for coronary heart disease (CHD). Early treatment of the disease improves the patient's prognosis. A screening consisting in the combination of TPA measurement and determining the maximum plaque thickness is recommended.


Better physical activity classification using smartphone acceleration sensor.
Obesity is becoming one of the serious problems for the health of worldwide population. Social interactions on mobile phones and computers via internet through social e-networks are one of the major causes of lack of physical activities. For the health specialist, it is important to track the record of physical activities of the obese or overweight patients to supervise weight loss control. In this study, acceleration sensor present in the smartphone is used to monitor the physical activity of the user. Physical activities including Walking, Jogging, Sitting, Standing, Walking upstairs and Walking downstairs are classified. Time domain features are extracted from the acceleration data recorded by smartphone during different physical activities. Time and space complexity of the whole framework is done by optimal feature subset selection and pruning of instances. Classification results of six physical activities are reported in this paper. Using simple time domain features, 99 % classification accuracy is achieved. Furthermore, attributes subset selection is used to remove the redundant features and to minimize the time complexity of the algorithm. A subset of 30 features produced more than 98 % classification accuracy for the six physical activities.


Crystal structure of human carbonic anhydrase XIII and its complex with the inhibitor acetazolamide.
The cytosolic isoform XIII is a recently discovered member of the human carbonic anhydrase (hCA, EC 126.96.36.199) family. It is selectively expressed among other tissues in the reproductive organs, where it may control pH and ion balance regulation, ensuring thus proper fertilization conditions. The authors report here the X-ray crystallographic structure of this isozyme in the unbound state and in complex with a classical sulfonamide inhibitor, namely acetazolamide. A detailed comparison of the obtained structural data with those already reported for other CA isozymes provides novel insights into the catalytic properties of the members of this protein family. On the basis of the inhibitory properties of acetazolamide against various cytosolic/transmembrane isoforms and the structural differences detected within the active site of the various CA isoforms, further prospects for the design of isozyme-specific CA inhibitors are here proposed.


Alkaline-earth (Be, Mg and Ca) bonds at the origin of huge acidity enhancements.
The interaction between alkaline-earth derivatives with the general formula X2M (X = H, F and Cl; M = Be, Mg and Ca) and a set of Lewis bases, including first and second-row hydrides, namely YHn (Y = O, N, F, S, P and Cl) hydrides, as well as other typical cyclic organic bases, such as aniline, 1H-1,2,3-triazole, 1H-tetrazole and phenylphosphine, was investigated using the G4 ab initio composite method. Contrary to what was expected, it was found that the interactions involving Mg and Ca derivatives were not necessarily weaker than those between beryllium bonds. The origin is two-fold: larger deformation of the interacting systems when Be-derivatives are involved and appearance of secondary non-covalent interactions in the formation of some of the Mg- and Ca-containing complexes. Hence, the dissociation of the latter complexes may require higher enthalpies than that of the Be complexes. These deformations are triggered by a significant redistribution of electron density of the two interacting moieties, which also result in dramatic changes in the reactivity of the interacting compounds and in particular in the intrinsic basicity of the Lewis bases investigated, to the point that conventional bases, such as ammonia or aniline, upon complexation with MCl2 (M = Be, Mg and Ca), become stronger Brønsted acids than phosphoric acid, whereas other bases, such as 1H-tetrazole, become stronger acids than perchloric acid.


Epidemiology of flavescence dorée in vineyards in northwestern Italy.
ABSTRACT A serious outbreak of flavescence dorée (FD) was reported in Piemonte, northwestern Italy, in 1998, and since then, the disease has compromised the economy of this traditional wine-growing area, even following the application of compulsory insecticide treatments to control Scaphoideus titanus, the vector of the causal phytoplasma. Affected vines show severe symptoms, varying according to the cultivar, and are rogued to reduce disease spread. Following winter and pruning, a previously affected vine may appear symptomless and free of phytoplasmas in its aerial as well as its root system, even by nested-polymerase chain reaction assays. Such plants are considered to be "recovered". Since 1998 homogenous data on the incidence of newly infected, healthy, or recovered plants productivity, presence of vectors, and treatment schedules have been collected in seven severely affected vineyards of southern Piemonte for 5 years (1999 to 2003). Infectivity and recovery rates were also calculated each year. From 1999 to 2003, the average number of healthy plants decreased and the numbers of recovered plants and those with symptoms increased. Productivity of recovered vines, although lower than that of healthy ones, was always higher than that of vines with symptoms and was not influenced by the time elapsed from date of recovery. The relationships between the ln-transformed number of vectors trapped in the vineyards the previous year and the infection and the recovery rates were fitted by an exponential (R(2) = 0.95) and an asymptotic (R(2) = 0.93) model, respectively.


Symptom recurrence following intermittent treatment in first-episode schizophrenia successfully treated for 2 years: a 3-year open-label clinical study.
An unanswered question in the management of schizophrenia is how long antipsychotic treatment should be continued after a single psychotic episode. In this study, we assessed the rates of symptom recurrence with intermittent treatment in patients with a first episode of DSM-IV-defined schizophrenia or related illness after 2 years of successful continuous treatment. We also investigated antecedents of recurrence, as well as demographic and baseline clinical predictors of early recurrence, and we compared the psychopathology of the recurrence episode with that of the first episode. Outpatients in an academic psychiatric hospital setting (single site) who had responded well in an open-label study with risperidone long-acting injection were recruited for this intermittent treatment trial, and those who participated had their treatment tapered and discontinued over a period of up to 6 weeks, with follow-up for 3 years or until reemergence of symptoms. Open-label treatment with oral risperidone and risperidone long-acting injection was immediately reinstituted in the event of recurrence of symptoms. The study was conducted between February 2004 and March 2010. The primary outcome measure was symptom recurrence rate at 3 years. Participants (N = 33) had a mean age ± SD of 28 ± 7.9 years and a mean baseline Positive and Negative Syndrome Scale total score ± SD of 44.8 ± 7.4 at study entry. Symptom recurrence rates were 79% at 12 months, 94% at 24 months, and 97% at 36 months. Onset of recurrence symptoms was fairly abrupt, and symptom severity returned to levels close to those of the first episode. No significant predictors of early recurrence were identified. Intermittent antipsychotic treatment, even after 2 years of successful treatment, may not be in the best interest of patients who have experienced a single psychotic episode. ClinicalTrials.gov identifier: NCT00378092.


Characterization of mitochondrial cytochromes P-450 from pig kidney and liver catalysing 26-hydroxylation of 25-hydroxyvitamin D3 and C27 steroids.
The properties of cytochrome P-450 from pig kidney mitochondria, catalysing 26-hydroxylation of 25-hydroxyvitamin D3 and C27 steroids [Postlind & Wikvall (1989) Biochem. Biophys. Res. Commun. 159, 1135-1140; Postlind (1990) Biochem. Biophys. Res. Commun. 168, 261-266], were compared with those of a 26-hydroxylating cytochrome P-450 from pig liver mitochondria. The liver enzyme was purified to a cytochrome P-450 content of 7.4 nmol/mg of protein and showed only one protein band with an apparent Mr of 53,000 upon SDS/PAGE. The cytochrome P-450 catalysed 26-hydroxylation of 25-hydroxyvitamin D3, cholesterol and 5 beta-cholestane-3 alpha, 7 alpha-diol at rates of 361, 1090 and 2065 pmol/min per nmol of cytochrome P-450. A monoclonal antibody against the purified liver mitochondrial cytochrome P-450 26-hydroxylase (cytochrome P-450(26] was prepared. After coupling to Sepharose, the antibody was able to bind to cytochrome P-450(26) from liver as well as from kidney mitochondria and to immunoprecipitate the 26-hydroxylase activity towards 25-hydroxyvitamin D3 and cholesterol when assayed in a reconstituted system. After SDS/PAGE and immunoblotting with the antibody, the cytochrome P-450(26) was detected in the purified liver and kidney preparations. These results indicate that similar species of cytochrome P-450 catalyse 26-hydroxylation of 25-hydroxyvitamin D3 and C27 steroids in liver and kidney mitochondria. The results with the monoclonal antibody together with the finding that cholesterol competitively inhibits the 26-hydroxylation of 25-hydroxyvitamin D3 further indicate that 26-hydroxylation of 25-hydroxyvitamin D3 and cholesterol is catalysed by the same species of cytochrome P-450 in each tissue. The N-terminal amino acid sequence of cytochrome P-450(26) in kidney mitochondria resembled that of pig kidney microsomal 25-hydroxylase active in 25-hydroxylation of vitamin D3 and C27 steroids, whereas the sequence of pig liver mitochondrial cytochrome P-450(26) differed from those of rabbit and rat liver mitochondrial 26-hydroxylases as well as from those of other hitherto isolated mammalian cytochromes P-450.


Vasopressin type 2 receptor V88M mutation: molecular basis of partial and complete nephrogenic diabetes insipidus.
Mutations in the type 2 vasopressin receptor gene (AVPR2) underlie X-linked recessive nephrogenic diabetes insipidus (NDI). Here, we report on a family with a mutation in AVPR2, c.262G>A (p.V88M). This recurrently identified mutation was previously shown to abolish AVPR2 function, yet in some affected members, urine osmolalities of up to 570 mosm/kg were observed. We detail the variable clinical phenotype and investigate its molecular basis. Retrospective analysis of clinical data and in vitro assessment of wild-type and V88M-mutant receptors. Clinical data were available on 6 patients. Four of these demonstrated a substantial increase in urinary concentration after 1-desamino[8-D-arginine] vasopressin, consistent with partial NDI, while 2 did not respond. In vitro analysis revealed a reduced cell surface expression and decreased binding affinity for arginine-vasopressin of the mutant receptor, leading to blunted signaling activity. Treatment with the pharmacological chaperone SR121463 enhanced cell surface expression. The V88M mutation is associated with phenotypical diversity, which may be explained by the fact that both the expression level and the hormone-binding affinity are affected by the mutation. Our results provide a rational basis for treatment trials with vasopressin analogues in combination with pharmacologic chaperones in patients with this recurrently identified mutation.


Alteration of polyunsaturated fatty acid status and metabolism in health and disease.
Essential polyunsaturated fatty acids (PUFA) cannot be synthesised in the body and must be ingested by food. A balanced intake of both n-6 and n-3 PUFA is essential for good health. PUFA are the basic constituents of phospholipid membranes and determine cellular membrane fluidity and modulate enzyme activities, carriers and membrane receptors. They are also precursors of active metabolites known collectively as eicosanoids (prostaglandins, prostacyclins, thromboxanes and leukotrienes) which regulate our cellular functions. Studies indicate that n-3 PUFA have anti-inflammatory, antithrombotic, antiarrhythmic actions and immuno-modulating properties. Erythrocyte fatty acid status is a reflection of dietary fat intake. It also explores PUFA metabolism and gives information about the integration of these fatty acids into cellular membranes. Thus, erythrocyte fatty acid analysis can detect PUFA insufficiencies and imbalances from the diet, but also metabolic abnormalities and lipid peroxidation. It can be helpful in the prevention and the control of chronic diseases in which PUFA alterations have been observed as coronary heart diseases, hypertension, cancer, diabetes, inflammatory and auto-immune disorders, atopic eczema, Alzheimer dementia, major depression, schizophrenia, multiple sclerosis, etc.


Proliferating cell nuclear antigen (PCNA) immunostaining--a prognostic factor in ovarian cancer?
The measurement of tumour cell proliferation is becoming increasingly recognised in defining prognostic groups. Proliferating cell nuclear antigen (PCNA) immunolocalisation can be used as an index of cell proliferation and may define the extent of departure from normal growth control. The monoclonal antibody PC10 stains PCNA in archival paraffin-embedded tissue. This study investigates its potential as a prognostic marker in early and advanced ovarian cancer. A three-stage immunoperoxidase technique was developed to detect the monoclonal antibody PC10. Archival paraffin-embedded tissue from 19 stage I ovarian tumours (13 malignant and six borderline) and 79 advanced (stage IIb-IV) ovarian tumours (patients entered into the Third North-West Thames Ovarian Cancer Trial) was immunostained with PC10. PC10 immunostaining was performed successfully in 91.8% of cases. The PC10 labelling index (PC10 LI) ranged from 1.5% to 88% with a mean value of 47.4%. Stage I borderline tumours had significantly lower PCNA labelling indexes than stage I malignant tumours (P < 0.048). In advanced disease there was an inverse correlation between PC10 and overall survival, and in those patients who underwent good debulking surgery (37 patients with disease < 2 cm diameter) a low PC10 value (< 36.5%) correlated with improved survival (log-rank trend test for survival, chi 2 = 5.75, P = 0.017). PCNA immunostaining defines a good prognostic subgroup in adequately debulked patients with ovarian cancer.


Biochemical kinetics of porcine cardiac subfragment-1. II. Pre-steady-state studies of the initial phosphate burst.
The actin dependence of the rate and magnitude of the initial phosphate burst was measured using both quench-flow and stopped-flow kinetic techniques. These studies revealed that even at high actin concentrations the magnitude of the phosphate burst was a significant fraction of the magnitude that exists in the absence of actin. Furthermore, it was shown that the rate of the burst rises rapidly as a function of the actin concentration. Detailed modeling with the four-state model revealed that if the predicted Vmax is constrained to be approximately equal to the extrapolated value (double reciprocal plot) and if the apparent dissociation constant of subfragment-1 to actin divided by the apparent activation constant of the actin-activated myosin ATPase activity (Kbinding/KATPase) is constrained to be considerably different from one, then the model is unable to simultaneously account for the ATPase activity and the rate and magnitude of the initial inorganic phosphate burst.


A Rare Form of Retinal Dystrophy Caused by Hypomorphic Nonsense Mutations in CEP290.
To identify the gene defect and to study the clinical characteristics and natural course of disease in a family originally diagnosed with oligocone trichromacy (OT), a rare congenital cone dysfunction syndrome. Extensive clinical and ophthalmologic assessment was performed on two siblings with OT and long-term follow up data were analyzed. Subsequently, whole exome sequencing (WES) and Sanger sequence analysis of CEP290 was performed in the two siblings. Additionally, the identified CEP290 mutations were analyzed in persons with achromatopsia (ACHM) (n = 23) and autosomal recessive or isolated cone dystrophy (CD; n = 145). In the first decade of life, the siblings were diagnosed with OT based on low visual acuity, photophobia, nystagmus, and absent cone response on electroretinography , but with normal color discrimination. Over time, the phenotype of OT evolved to a progressive degenerative disease without any CEP290-associated non-ocular features. In both siblings, two nonsense mutations (c.451C>T; p.(Arg151*) and c.4723A>T; p.(Lys1575*)) in CEP290 were found. Previously, p.(Arg151*) was demonstrated to induce nonsense-mediated alternative splicing events leading to intact open reading frames of the resulting mRNA products (p.(Leu148_Glu165del) and p.(Leu148_Lys172del)). mRNA analysis for p.(Lys1575*) confirmed a suspected hypomorphic character, as exon 36 skipping was observed in a small fraction of CEP290 mRNA, resulting in a 36 aa in-frame deletion (p.(Glu1569_Trp1604del)). No additional cases carrying these variants were identified in the ACHM and CD cohorts. Compound heterozygous hypomorphic mutations in CEP290 may lead to a rare form of cone-dominated retinal dystrophy, a novel phenotype belonging to the CEP290-associated spectrum of ciliopathies. These findings provide insight into the effect of CEP290 mutations on the clinical phenotype.


Effects of live yeast Saccharomyces cerevisiae on fermentation parameters and microbial populations of rumen, total tract digestibility of diet nutrients and on the in situ degradability of alfalfa hay in Iranian Chall sheep.
The effects of live yeast Saccharomyces cerevisiae (LYSC, strain Sc 47) on rumen fermentation and microbial populations, digestibility of nutrients, degradability of alfalfa hay and on the performances of sheep were investigated in two experiments. In both experiments, animals in treatment groups were received 0, 2.5, 5 and 7.5 g of LYSC per sheep per day and were defined as control, 2.5, 5.0 and 7.5 g LYSC treatment groups, respectively. In the first experiment, four fistulated Chall sheep (49 +/- 0.5 kg BW) were kept in individual metabolic crates under a 4x4 Latin square design and fed a Total Mixed Ration (TMR) containing of barely (48%), wheat bran (16%), shelled corn (5%), mineral-vitamin mix (1%) and of alfalfa hay (30%). In the second experiment, 28 sheep (48 +/- 0.5 kg BW) were assigned into four treatment groups under a complete randomized design and fed a TMR containing of barely (60%), wheat bran (7%), cottonseed meal (2%), mineral-vitamin mix (1%) and of alfalfa hay (15%) and wheat straw (15%). The highest and the lowest ruminal pH values (p < 0.01) were recorded for sheep in 2.5 g LYSC and control groups, respectively. At 3 h post-feeding, the total VFA of rumen fluid was increased (p < 0.01) from 91.26 to 103.34 mmol L(-1) in control vs. 2.5 g LYSC groups. The ruminal NH3-N of sheep was decreased (p < 0.01) from 159.63 to 128.90 mg L(-1) in control vs. 2.5 g LYSC groups. Bacterial populations of rumen fluids were differed from 14 to 43% in treatment groups although the differences were not significant. Compared to the other groups, voluntary feed intake was higher (p < 0.01) in 5.0 g LYSC group and this was ended to an inappropriate feed to gain ratio in this group. It can be concluded that the use of LYSC at a level of 2.5 g per sheep per day could improve the ruminal fermentation and resulted in a relatively better performances in Chall sheep.


Endothelin-1 increases superoxide production in human coronary artery bypass grafts.
Endothelin-1 (ET-1) has been shown to increase endothelial superoxide (O(2)(-)) production in experimental animal models. It is unclear whether ET-1 increases O(2)(-) production in humans. We sought to elucidate whether ET-1 increases O(2)(-) production in human vessels and to identify the mechanism behind this effect. Segments of internal mammary artery (IMA) and human saphenous vein (HSV) were harvested from 90 patients undergoing elective coronary artery bypass graft surgery. Paired vessel rings were incubated in the presence and absence of ET-1 (10(-10)M), the ET(A) receptor antagonist BQ123 alone, or in combination with the ET(B) receptor antagonist BQ788 (dual BQ) and known inhibitors of sources of O(2)(-) and further analysed for O(2)(-) production using lucigenin-enhanced chemiluminescence and DHE fluorescence. ET-1 increased O(2)(-) production in both IMA (2.6 ± 1.5 vs. 1.4 ± 0.8 relative light units/s/mg tissue (RLU); n=33; p < 0.0001) and HSV (1.4 ± 0.8 vs. 1.1 ± 0.6 RLU; n=24; p<0.05). The increase in O(2)(-)production induced by ET-1 in IMA was inhibited by co-incubation with dual BQ (p < 0.05; n=15) and BQ123 (p<0.05; n = 17). Of known O(2)(-) inhibitors, only incubation with Tiron and diphenyleneiodonium resulted in a significant reduction in ET-mediated O(2)(-) production. ET-1 increases O(2)(-) production especially in human arteries and less so in veins from patients with coronary artery disease via a receptor-dependent pathway involving a flavin dependent enzyme which is likely to be NADPH oxidase. Production of O(2)(-) may be an important factor underlying the negative effects of ET-1 on vascular function such as impairment of endothelium-dependent vasodilatation and pro-inflammatory effects.


Predictive Value of Coronary Artery Calcium Score Categories for Coronary Events Versus Strokes: Impact of Sex and Race: MESA and DHS.
Coronary artery calcium (CAC) predicts atherosclerotic cardiovascular disease (ASCVD) events, inclusive of coronary heart disease (CHD) and stroke, and is a decision-making aid for primary prevention. The predictive value of CAC categories for CHD and stroke separately and across sex and race groups of an asymptomatic population is unclear. White, Black, and Hispanic participants of MESA (Multi-Ethnic Study of Atherosclerosis) and DHS (Dallas Heart Study) underwent CAC measurement at enrollment and were followed for incident ASCVD events. Ten-year CHD-to-stroke incidence ratios across CAC score categories 0, 1 to 99, and ≥100 were assessed. Associations of CAC with incident CHD and stroke events were evaluated using multivariable-adjusted Cox models and multiplicative interactions of CAC with sex/race were tested. Among 7042 participants (mean age, 57 years, 54% women, 36% Black, 23% Hispanic, 49% CAC=0, 19% CAC ≥100), 574 incident ASCVD events (333 CHD and 241 stroke) were observed over 12.3-year follow-up. Ten-year CHD-to-stroke incidence ratio increased significantly across CAC categories in men, women, Whites, Blacks, and Hispanics (all P<0.001). High CAC burden (score ≥100) was independently associated with ASCVD and CHD risk in all groups and with stroke risk in the overall cohort and Blacks. No sex- or race-based CAC interactions for ASCVD, CHD, and stroke events were observed. Adding CAC to a traditional risk factor model improved risk discrimination and reclassification for CHD but not for stroke events. In 2 population-based cohorts of asymptomatic individuals, 10-year CHD-to-stroke incidence ratio was higher with increasing CAC score categories across sex and race groups, and CAC was consistently a better predictor of CHD than stroke. High CAC burden comparably associated with ASCVD risk across sex and race groups.


Roles of retinoids and their nuclear receptors in the development and prevention of upper aerodigestive tract cancers.
Vitamin A analogs (retinoids) suppress oral and lung carcinogenesis in animal models and prevent the development of second primary tumors in head, neck, and lung cancer patients. These effects result from changes in the expression of genes that regulate cell growth and differentiation. Retinoic acid receptors (RARs; -alpha, -beta, and -gamma) and retinoid X receptors (RXRs; -alpha, -beta, and, -gamma) are retinoid-activated transcription factors, which mediate effects of retinoids on gene expression. Therefore, alterations in receptor expression or function could interfere with the retinoid signaling pathway and thereby enhance cancer development. We found that the expression of RAR beta was suppressed in more than 50% of oral and lung premalignant lesions in individuals without cancer and in dysplastic lesions adjacent to cancer and in malignant oral and lung carcinomas. The expression of the other receptors was not different among normal, dysplastic, and malignant oral tissues. However, the expression of RAR gamma and RXR beta was somewhat decreased in lung cancers. These results show that RAR beta expression is lost at early stages of carcinogenesis in the aerodigestive tract and support the hypothesis that the loss of RAR beta expression may facilitate the development of some of these cancers.


Tumor-dependent kinetics of partial pressure of oxygen fluctuations during air and oxygen breathing.
The primary purpose of this study was to examine the kinetics of partial pressure of oxygen (pO2) fluctuations in fibrosarcoma (FSA) and 9L tumors under air and O2 breathing conditions. The overall hypothesis was that key factors relating to oxygen tension fluctuations would vary between the two tumor types and as a function of the oxygen content of the breathing gas. To assist in the interpretation of the temporal data, spatial pO2 distributions were measured in 10 FSA and 8 9L tumors transplanted into the subcutis of the hind leg of Nembutal-anesthetized (50 mg/kg) Fischer 344 rats. Recessed-tip oxygen microelectrodes were inserted into the tumor, and linear pO2 measurements were recorded in 50-microm steps along a 3-mm path, and blood pressure was simultaneously measured via femoral arterial access. Additionally, pO2 was measured at a single location for 90 to 120 minutes in FSA (n=11) or 9L tumors (n=12). Rats were switched from air to 100% O2 breathing after 45 minutes. Temporal pO2 records were evaluated for their potential radiobiological significance by assessing the number of times they crossed a 10-mm-Hg threshold. In addition, the data were subjected to Fourier analysis for air and O2 breathing. FSA and 9L tumors had spatial median pO2 measurements of 4 and 1 mm Hg, respectively. 9L had more low pO2 measurements < or =2.5 mm Hg than did FSA, whereas between 2.5 and 10 mm Hg this pattern was reversed. Pimonidazole staining patterns in FSA and 9L tumors supported these results. Temporal pO2 instability was observed in all experiments during air and O2 breathing. Threshold analyses indicated that the 10 mm Hg threshold was crossed 2 to 5 times per hour, independent of tumor type. However, the magnitude of 9L pO2 fluctuations was approximately eight times greater than FSA fluctuations, as assessed with Fourier transform analysis (Wilcoxon, P < 0.005). O2 breathing significantly increased median pO2 in FSA from 3 to 8 mm Hg (P < 0.005) and caused a significant increase in frequency and magnitude of pO2 fluctuations. One hundred percent O2 breathing had no effect on 9L tumor pO2, and it decreased the magnitude of pO2 fluctuations with borderline significance. These results show that these two tumors differ significantly with respect to spatial and temporal oxygenation conditions under air and O2 breathing. Fluctuations of pO2 of the type reported herein are predicted to significantly affect radiotherapy response and could be a source for genetic instability, increased angiogenesis, and metastases.


The role of the cerebellum in sub- and supraliminal error correction during sensorimotor synchronization: evidence from fMRI and TMS.
Our ability to interact physically with objects in the external world critically depends on temporal coupling between perception and movement (sensorimotor timing) and swift behavioral adjustment to changes in the environment (error correction). In this study, we investigated the neural correlates of the correction of subliminal and supraliminal phase shifts during a sensorimotor synchronization task. In particular, we focused on the role of the cerebellum because this structure has been shown to play a role in both motor timing and error correction. Experiment 1 used fMRI to show that the right cerebellar dentate nucleus and primary motor and sensory cortices were activated during regular timing and during the correction of subliminal errors. The correction of supraliminal phase shifts led to additional activations in the left cerebellum and right inferior parietal and frontal areas. Furthermore, a psychophysiological interaction analysis revealed that supraliminal error correction was associated with enhanced connectivity of the left cerebellum with frontal, auditory, and sensory cortices and with the right cerebellum. Experiment 2 showed that suppression of the left but not the right cerebellum with theta burst TMS significantly affected supraliminal error correction. These findings provide evidence that the left lateral cerebellum is essential for supraliminal error correction during sensorimotor synchronization.


Intranasal vaccination against angiotensin II type 1 receptor and pneumococcal surface protein A attenuates hypertension and pneumococcal infection in rodents.
To combat global increases in the prevalence of lifestyle-related diseases and concomitant infectious diseases, we aimed to develop an innovative intranasal vaccine that simultaneously targets both hypertension and pneumonia, is not given by invasive injection, and offers prolonged therapeutic effect and reduced frequency of administration. Angiotensin II type 1 receptor-pneumococcal surface protein A (AT1R-PspA) vaccine, consisting of a cationic nanometer-sized hydrogel incorporating AT1R partial peptide conjugated with PspA and cyclic diguanylate monophosphate adjuvant, was created and given intranasally to spontaneously hypertensive rats (SHRs). Antigen-specific antibodies and blood pressure were examined to evaluate immune responses and the antihypertensive effect of the vaccine. To examine the protective effect of antibodies induced by vaccination on pneumococcal infection, sera obtained from immunized SHRs were incubated with a lethal dose of Streptococcus pneumoniae and then administered to mice. Five doses of AT1R-PspA nasal-vaccine-induced AT1R-specific serum IgG antibody production and attenuated the development of hypertension in SHRs in the long term. Both in-vitro and in-vivo studies revealed that responses to angiotensin II were suppressed in vaccinated rats. Mice passively immunized with sera obtained from AT1R-PspA-vaccinated SHRs were protected from lethal pneumococcal infection. Intranasal immunization with AT1R-PspA vaccine has the potential to simultaneously attenuate the development of hypertension and protect from lethal pneumococcal infection.


Sex differences in immune responses to SARS-CoV-2 that underlie disease outcomes.
A growing body of evidence indicates sex differences in the clinical outcomes of coronavirus disease 2019 (COVID-19)1-4. However, whether immune responses against SARS-CoV-2 differ between sexes, and whether such differences explain male susceptibility to COVID-19, is currently unknown. In this study, we examined sex differences in viral loads, SARS-CoV-2-specific antibody titers, plasma cytokines, as well as blood cell phenotyping in COVID-19 patients. By focusing our analysis on patients with mild to moderate disease who had not received immunomodulatory medications, our results revealed that male patients had higher plasma levels of innate immune cytokines and chemokines including IL-8, IL-18, and CCL5, along with more robust induction of non-classical monocytes. In contrast, female patients mounted significantly more robust T cell activation than male patients during SARS-CoV-2 infection, which was sustained in old age. Importantly, we found that a poor T cell response negatively correlated with patients' age and was predictive of worse disease outcome in male patients, but not in female patients. Conversely, higher innate immune cytokines in female patients associated with worse disease progression, but not in male patients. These findings reveal a possible explanation underlying observed sex biases in COVID-19, and provide important basis for the development of sex-based approach to the treatment and care of men and women with COVID-19.


The utility of social capital in research on health determinants.
Social capital has become a popular subject in the literature on determinants of health. The concept of social capital has been used in the sociological, political science, and economic development literatures, as well as in the health inequalities literature. Analysis of its use in the health inequalities literature suggests that each theoretical tradition has conceptualized social capital differently. Health researchers have employed a wide range of social capital measures, borrowing from several theoretical traditions. Given the wide variation in these measures and an apparent lack of consistent theoretical or empirical justification for their use, conclusions about the likely role of "social capital" on population health may be overstated or even misleading. Elements of a research agenda are proposed to further elucidate the potential role of factors currently subsumed under the rubric of "social capital."


Clinical outcomes after intentional replantation of permanent teeth: A systematic review.
This study aimed to systematically assess the outcomes of intentional replantation (IR) of teeth and to determine the survival rate, success rate, and prognostic factors related to the treatment. A search was conducted for all relevant English language articles published from January 2000 to October 2017. The search terms included "intentional replantation" and "teeth" according to the inclusion criteria. The Methodological Index for Non-randomized Studies (MINORS) was used to assess the methodological quality of included studies. Twelve studies were identified as relevant for the systematic review. In total, 896 patients with 905 teeth were examined for intentional teeth replantation. The success rate was greater than 90% in four studies (33.33%) and between 70% and 80% in five studies. At short-term follow-up (<6 months), the survival rate was approximately 90% in four studies. At longer-term follow-up (>36 months), the survival rates of teeth were slightly reduced in four studies, and tended to be stable after 48 months. In conclusion, the long-term success and survival rate of IR are likely dependent upon short extraoral time, reduced pocket depth, type of tooth, type of root-end filling material, and the prevention of atraumatic tooth root damage.


Tumor pH and Protein Concentration Contribute to the Signal of Amide Proton Transfer Magnetic Resonance Imaging.
Abnormal pH is a common feature of malignant tumors and has been associated clinically with suboptimal outcomes. Amide proton transfer magnetic resonance imaging (APT MRI) holds promise as a means to noninvasively measure tumor pH, yet multiple factors collectively make quantification of tumor pH from APT MRI data challenging. The purpose of this study was to improve our understanding of the biophysical sources of altered APT MRI signals in tumors. Combining in vivo APT MRI measurements with ex vivo histological measurements of protein concentration in a rat model of brain metastasis, we determined that the proportion of APT MRI signal originating from changes in protein concentration was approximately 66%, with the remaining 34% originating from changes in tumor pH. In a mouse model of hypopharyngeal squamous cell carcinoma (FaDu), APT MRI showed that a reduction in tumor hypoxia was associated with a shift in tumor pH. The results of this study extend our understanding of APT MRI data and may enable the use of APT MRI to infer the pH of individual patients' tumors as either a biomarker for therapy stratification or as a measure of therapeutic response in clinical settings. SIGNIFICANCE: These findings advance our understanding of amide proton transfer magnetic resonance imaging (APT MRI) of tumors and may improve the interpretation of APT MRI in clinical settings.


Modular time division multiplexer: Efficient simultaneous characterization of fast and slow transients in multiple samples.
A modular time division multiplexer (MTDM) device is introduced to enable parallel measurement of multiple samples with both fast and slow decay transients spanning from millisecond to month-long time scales. This is achieved by dedicating a single high-speed measurement instrument for rapid data collection at the start of a transient, and by multiplexing a second low-speed measurement instrument for slow data collection of several samples in parallel for the later transients. The MTDM is a high-level design concept that can in principle measure an arbitrary number of samples, and the low cost implementation here allows up to 16 samples to be measured in parallel over several months, reducing the total ensemble measurement duration and equipment usage by as much as an order of magnitude without sacrificing fidelity. The MTDM was successfully demonstrated by simultaneously measuring the photoconductivity of three amorphous indium-gallium-zinc-oxide thin films with 20 ms data resolution for fast transients and an uninterrupted parallel run time of over 20 days. The MTDM has potential applications in many areas of research that manifest response times spanning many orders of magnitude, such as photovoltaics, rechargeable batteries, amorphous semiconductors such as silicon and amorphous indium-gallium-zinc-oxide.


Human neutrophils express immunoglobulin E (IgE)-binding proteins (Mac-2/epsilon BP) of the S-type lectin family: role in IgE-dependent activation.
It has been suggested that neutrophils may be involved in the late-phase reaction of immunoglobulin E (IgE)-dependent hypersensitivity states. However, the identity of neutrophil-associated molecules inducing the release of mediators remains unclear. In this report, we demonstrate that human neutrophils from normal donors or from patients with inflammatory disorders could bind myeloma IgE proteins, especially after desialylation. Northern blot, immunoprecipitation, and flow cytometry analyses revealed that neutrophils did not express Fc epsilon RII/CD23, but rather Mac-2/epsilon binding protein (BP), belonging to the S-type lectin family. Similarly to IgA used as positive control, myeloma IgE proteins, as well as polyclonal IgE antibodies with or without antibody specificity, were both capable of inducing a neutrophil respiratory burst. Anti-Mac-2 but not anti-CD23 mAb strongly decreased the IgE-dependent activation of neutrophils, induced either by the specific antigen or by anti-IgE antibodies. These findings open new perspectives on the functional role of neutrophils in IgE-associated diseases including allergic states or parasitic infections.


Lumbar disc herniation. A controlled, prospective study with ten years of observation.
Two hundred eighty patients with herniated lumbar discs, verified by radiculography, were divided into three groups. One group, which mainly will be dealt with in this paper, consisted of 126 patients with uncertain indication for surgical treatment, who had their therapy decided by randomization which permitted comparison between the results of surgical and conservative treatment. Another group comprising 67 patients had symptoms and signs that beyond doubt, required surgical therapy. The third group of 87 patients was treated conservatively because there was no indication for operative intervention. Follow-up examinations in the first group were performed after one, four, and ten years. The controlled trial showed a statistically significant better result in the surgically treated group at the one-year follow-up examination. After four years the operated patients still showed better results, but the difference was no longer statistically significant. Only minor changes took place during the last six years of observation.