The efficacy of human placenta as a source of the universal feeder in human and mouse pluripotent stem cell culture.
The use of a mouse embryonic fibroblast (MEF) feeder for culture of embryonic stem cells (ESCs) is a widely accepted method, regardless of the ESCs' origin and type. In this study, we performed the undifferentiated propagation of human ES cell lines (hESCs, H1, and HSF6) and mouse ES cell lines (mESCs, D3, and CE3), which were previously maintained on an MEF feeder, using human placenta-derived fibroblast-like cell (HPC) feeders originated from chorionic villi of women who had undergone therapeutic abortion due to known maternal disease that is aggravated by pregnancy. Moreover, we tried to introduce the HPC feeder for the establishment of inducible pluripotent stem cells (iPSCs) from human placental mesenchymal stem cells (MSCs). On the HPC feeder we were able to propagate ESCs and iPSCs colonies as an undifferentiated state up to the 50th passage and 20th passage, respectively. Maintenance of undifferentiated ESCs was identified by the expression of ALP, SSEA-1, SSEA-4, TRA-81, TRA-60, Oct-4, Nanog, or Rex-1. Also, addition of leukemia inhibitory factor was not required for undifferentiated propagation of mESCs on the HPC feeder. The efficiency and expression of three germ layer markers of embryoid bodies (EBs) from ESCs were satisfactory in both the MEF and HPC group. EBs formed from iPSCs were scant, and differentiation to the three germ layers was identifiable by reverst transcription-polymerase chain reactio (RT-PCR) only in the HPC group. In conclusion, the HPC feeder can efficiently support the undifferentiated propagation of hESCs, mESCs, and iPSCs, suggesting that human placenta may be a useful source of universal feeder cells for hESC, mESC, and iPSC culture.


Proprotein convertase subtilisin/kexin type 9: a new target molecule for gene therapy.
Proprotein convertase subtilisin/kexin type 9 (PCSK9) has emerged as a novel target for controlling plasma levels of low-density lipoprotein cholesterol (LDL-C) and decreasing the risk of cardiovascular diseases. At present it is clear that the major classes of commonly prescribed lipid-lowering medications increase serum PCSK9 levels and fail to protect a significant percentage of patients from cardiovascular events. Therefore development of new LDL-C lowering medications that either do not increase circulating PCSK9 levels or work through inhibition of PCSK9 expression and protease activity is a highly desirable approach to overcome hypercholesterolemia. Since there are several agents which are being evaluated in human preclinical and clinical trials, this review summarizes current therapeutic strategies targeting PCSK9, including specific antibodies, antisense oligonucleotides, small interfering RNAs (siRNAs) and other small-molecule inhibitors.


Physician-Assisted Suicide: Why Neutrality by Organized Medicine Is Neither Neutral Nor Appropriate.
It has been proposed that medical organizations adopt neutrality with respect to physician-assisted suicide (PAS), given that the practice is legal in some jurisdictions and that membership is divided. We review developments in end-of-life care and the role of medical organizations with respect to the legalization of PAS since the 1990s. We argue that moving from opposition to neutrality is not ethically neutral, but a substantive shift from prohibited to optional. We argue that medical organizations already oppose many practices that are legal in many jurisdictions, and that unanimity among membership has not been required for any other clinical or ethical policy positions. Moreover, on an issue so central to the meaning of medical professionalism, it seems important for organized medicine to take a stand. We subsequently review the arguments in favor of PAS (arguments from autonomy and mercy, and against the distinction between killing and allowing to die (K/ATD)) and the arguments against legalization (the limits of autonomy, effects on the patient-physician relationship, the meaning of healing, the validity of the K/ATD distinction, the social nature of suicide, the availability of alternatives, the propensity for incremental extension, and the meaning of control). We conclude that organized medicine should continue its opposition to PAS.


Minimizing carry-over PCR contamination in expanded CAG/CTG repeat instability applications.
Expanded CAG/CTG repeats underlie the aetiology of 14 neurological and neuromuscular disorders. The size of the repeat tract determines in large part the severity of these disorders with longer tracts causing more severe phenotypes. Expanded CAG/CTG repeats are also unstable in somatic tissues, which is thought to modify disease progression. Routine molecular biology applications involving these repeats, including quantifying their instability, are plagued by low PCR yields. This leads to the need for setting up more PCRs of the same locus, thereby increasing the risk of carry-over contamination. Here we aimed to reduce this risk by pre-treating the samples with a Uracil N-Glycosylase (Ung) and using dUTP instead of dTTP in PCRs. We successfully applied this method to the PCR amplification of expanded CAG/CTG repeats, their sequencing, and their molecular cloning. In addition, we optimized the gold-standard method for measuring repeat instability, small-pool PCR (SP-PCR), such that it can be used together with Ung and dUTP-containing PCRs, without compromising data quality. We performed SP-PCR on myotonic-dystrophy-derived samples containing an expansion as large as 1000 repeats, demonstrating the applicability to clinically-relevant material. Thus, we expect the protocols herein to be applicable for molecular diagnostics of expanded repeat disorders.


Early Holocene chicken domestication in northern China.
Chickens represent by far the most important poultry species, yet the number, locations, and timings of their domestication have remained controversial for more than a century. Here we report ancient mitochondrial DNA sequences from the earliest archaeological chicken bones from China, dating back to ∼ 10,000 B.P. The results clearly show that all investigated bones, including the oldest from the Nanzhuangtou site, are derived from the genus Gallus, rather than any other related genus, such as Phasianus. Our analyses also suggest that northern China represents one region of the earliest chicken domestication, possibly dating as early as 10,000 y B.P. Similar to the evidence from pig domestication, our results suggest that these early domesticated chickens contributed to the gene pool of modern chicken populations. Moreover, our results support the idea that multiple members of the genus Gallus, specifically Gallus gallus and Gallus sonneratii contributed to the gene pool of the modern domestic chicken. Our results provide further support for the growing evidence of an early mixed agricultural complex in northern China.


Flavocytochrome P450 BM3 mutant W1046A is a NADH-dependent fatty acid hydroxylase: implications for the mechanism of electron transfer in the P450 BM3 dimer.
Bacillus megaterium P450 BM3 (BM3) is a P450/P450 reductase fusion enzyme, where the dimer is considered the active form in NADPH-dependent fatty acid hydroxylation. The BM3 W1046A mutant was generated, removing an aromatic "shield" from its FAD isoalloxazine ring. W1046A BM3 is a catalytically active NADH-dependent lauric acid hydroxylase, with product formation slightly superior to the NADPH-driven enzyme. The W1046A BM3 K(m) for NADH is 20-fold lower than wild-type BM3, and catalytic efficiency of W1046A BM3 with NADH and NADPH are similar in lauric acid oxidation. Wild-type BM3 also catalyzes NADH-dependent lauric acid hydroxylation, but less efficiently than W1046A BM3. A hypothesis that W1046A BM3 is inactive [15] helped underpin a model of electron transfer from FAD in one BM3 monomer to FMN in the other in order to drive fatty acid hydroxylation in native BM3. Our data showing W1046A BM3 is a functional fatty acid hydroxylase are consistent instead with a BM3 catalytic model involving electron transfer within a reductase monomer, and from FMN of one monomer to heme of the other [12]. W1046A BM3 is an efficient NADH-utilizing fatty acid hydroxylase with potential biotechnological applications.


ARIA-EAACI statement on severe allergic reactions to COVID-19 vaccines - An EAACI-ARIA Position Paper.
Further to the approval of the Coronavirus disease 2019 (COVID-19) vaccine BNT162b2, several severe anaphylaxis cases occured within the first few days of public vaccination. An investigation is taking place to understand the cases and their triggers. The vaccine will be administered to a large number of individuals worldwide and there are raising concerns that severe adverse events might occur. With the current information, the European Academy of Allergy and Clinical Immunology (EAACI) states its position for the following preliminary recommendations that are to be revised as soon as more data emerge. To minimize the risk of severe allergic reactions in vaccinated individuals, it is urgently required to understand the specific nature of the reported severe allergic reactions, including the background medical history of the individuals affected and the mechanisms involved. To achieve this goal, all clinical and laboratory information should be collected and reported. Mild and moderate allergic patients should not be excluded from the vaccine as this could have a significant impact on reaching the goal of population immunity. Healthcare practitioners vaccinating against COVID-19 are required to be sufficiently prepared to recognize and treat anaphylaxis properly with the ability to administer adrenaline. Further to vaccine administration, a mandatory observation period of at least 15 minutes should be followed for all individuals. The current data have not shown any higher risk for patients suffering from allergic rhinitis or asthma, and this message should be clearly stated by physicians to enable our patients to trust the vaccine. More than 30% of the population suffers from allergic diseases and the benefit of the vaccination clearly outweighs the risk of severe COVID-19 development.


Medical illness and the risk of suicide in the elderly.
Suicide is a leading cause of death, and rates are especially high among the elderly. Medical illnesses may predispose to suicide, but few controlled studies have examined the association between specific diseases and suicide. We explored the relationship between treatment for several illnesses and the risk of suicide in elderly patients using a population-based approach. All Ontario residents 66 years or older who committed suicide between January 1, 1992, and December 31, 2000, were identified from provincial coroners' records. Their prescription records during the preceding 6 months were compared with those of living matched controls (1:4) to determine the presence or absence of 17 illnesses potentially associated with suicide. During the 9-year study period, we identified 1354 elderly patients who died of suicide. The most common mechanisms involved firearms (28%), hanging (24%), and self-poisoning (21%). Specific illnesses associated with suicide included congestive heart failure (odds ratio [OR], 1.73; 95% confidence interval [CI], 1.33-2.24), chronic obstructive lung disease (OR, 1.62; 95% CI, 1.37-1.92), seizure disorder (OR, 2.95; 95% CI, 1.89-4.61), urinary incontinence (OR, 2.02; 95% CI, 1.29-3.17), anxiety disorders (OR, 4.65; 95% CI, 4.07-5.32), depression (OR, 6.44; 95% CI, 5.45-7.61), psychotic disorders (OR, 5.09; 95% CI, 3.94-6.59), bipolar disorder (OR, 9.20, 95% CI, 4.38-19.33), moderate pain (OR, 1.91; 95% CI, 1.66-2.20), and severe pain (OR, 7.52; 95% CI, 4.93-11.46). Treatment for multiple illnesses was strongly related to a higher risk of suicide. Almost half the patients who committed suicide had visited a physician in the preceding week. Many common illnesses are independently associated with an increased risk of suicide in the elderly. The risk is greatly increased among patients with multiple illnesses. These data may help clinicians to identify elderly patients at risk of suicide and open avenues for prevention.


Regional variation in use of immediate breast reconstruction after mastectomy for breast cancer in England.
English national guidelines recommend that breast reconstruction is made available to women with breast cancer undergoing mastectomy. We examined the use of immediate reconstruction (IR) across English Cancer Networks, who are responsible for the regional organisation of cancer services and ensuring equitable access to treatment. We analysed Hospital Episodes Statistics data for all women with breast cancer who underwent mastectomy in the English NHS between April 2006 and February 2009. IR rates were calculated for the 30 Networks. Multivariable logistic regression was used to adjust the rates for patient age, comorbidity, ethnicity and socioeconomic deprivation. Of 44 837 mastectomy patients, 7375 (16.5%) underwent IR. The IR rate was highest in women under 50 years (32.7%) and lowest in women aged 70 years or over (1.5%), and was lower in women with more comorbidities. Unadjusted IR rates varied from 8.4% to 31.9% among the 30 Networks (p<0.001). Adjusting for their patient characteristics did not appreciably reduce Network-level variation, with adjusted IR rates still ranging from 8.0% to 29.4% (p<0.001). The risk-model also suggested that non-white women and those from more deprived areas were less likely to undergo immediate reconstruction. There is substantial regional variation in immediate reconstruction use in England that is not explained by the characteristics of the local patient population. English Cancer Networks should act to reduce this variation. They should also examine why rates of reconstruction differ between particular patient groups.


Impact of heterogeneity within cultured cells on bacterial invasion: analysis of Pseudomonas aeruginosa and Salmonella enterica serovar typhi entry into MDCK cells by using a green fluorescent protein-labelled cystic fibrosis transmembrane conductance regulator receptor.
The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride ion channel that also serves as a receptor for entry of Pseudomonas aeruginosa and Salmonella enterica serovar Typhi into epithelial cells. To evaluate heterogeneity in CFTR protein expression in cultured cells and the effect of heterogeneity on internalization of different P. aeruginosa and serovar Typhi strains, we used two-color flow cytometry and confocal laser microscopy to study bacterial uptake by Madin-Darby canine kidney (MDCK) type I epithelial cells stably expressing a green fluorescent protein (GFP)-CFTR fusion construct (MDCK-GFP-CFTR cells). We found a strong correlation between cell size and GFP-CFTR protein expression, with 60 to 70% of cells expressing low levels of GFP-CFTR protein, 20 to 30% expressing intermediate levels, and <10% expressing high levels. The cells were sorted into low-, intermediate-, or high-level producers of CFTR protein; in vitro growth of each sorted population yielded the same distribution of CFTR protein expression as that in the original population. Cells expressing either low or high levels of CFTR protein internalized bacteria poorly; maximal bacterial uptake occurred in the cells expressing intermediate levels of CFTR protein. Treatment of MDCK cells with sodium butyrate markedly enhanced the production of CFTR protein without increasing cell size; butyrate treatment also increased the proportion of cells with internalized bacteria. However, there were fewer bacteria per butyrate-treated cell and, for P. aeruginosa, there was an overall decrease in the total level of bacterial uptake. The most highly ingested bacterial strains were internalized by fewer total MDCK-GFP-CFTR cells, indicating preferential bacterial uptake by a minority of epithelial cells within a given culture. Confocal fluorescence microscopy showed that P. aeruginosa and serovar Typhi induced cytoplasmic accumulation of CFTR protein close to the plasma membrane where the bacteria were adherent. These results show that within a population of MDCK-GFP-CFTR cells, there are cells with markedly different abilities to ingest bacteria via CFTR, the majority of the P. aeruginosa and serovar Typhi cells are ingested by the one-fourth to one-third of the cells that exhibit an intermediate size and level of CFTR protein expression, and overexpression of the CFTR receptor does not increase total bacterial uptake but rather allows more epithelial cells to ingest fewer total bacteria.


Analysis of the scalability of diffraction-limited fiber lasers and amplifiers to high average power.
We analyze the scalability of diffraction-limited fiber lasers considering thermal, non-linear, damage and pump coupling limits as well as fiber mode field diameter (MFD) restrictions. We derive new general relationships based upon practical considerations. Our analysis shows that if the fiber's MFD could be increased arbitrarily, 36 kW of power could be obtained with diffraction-limited quality from a fiber laser or amplifier. This power limit is determined by thermal and non-linear limits that combine to prevent further power scaling, irrespective of increases in mode size. However, limits to the scaling of the MFD may restrict fiber lasers to lower output powers.


The effect of Jun dimerization on neurite outgrowth and motif binding.
Axon regeneration is a necessary step toward functional recovery after spinal cord injury. The AP-1 transcription factor c-Jun has long been known to play an important role in directing the transcriptional response of Dorsal Root Ganglion (DRG) neurons to peripheral axotomy that results in successful axon regeneration. Here we performed ChIPseq for Jun in mouse DRG neurons after a sciatic nerve crush or sham surgery in order to measure the changes in Jun's DNA binding in response to peripheral axotomy. We found that the majority of Jun's injury-responsive changes in DNA binding occur at putative enhancer elements, rather than proximal to transcription start sites. We also used a series of single polypeptide chain tandem transcription factors to test the effects of different Jun-containing dimers on neurite outgrowth in DRG, cortical and hippocampal neurons. These experiments demonstrated that dimers composed of Jun and Atf3 promoted neurite outgrowth in rat CNS neurons as well as mouse DRG neurons. Our work provides new insight into the mechanisms underlying Jun's role in axon regeneration.


A theoretical perspective of the enhanced photocatalytic properties achieved by forming tetragonal ZnS/ZnSe hetero-bilayer.
An efficient and economical way to tune the properties of two-dimensional (2D) materials is by forming van der Waals (vdW) hetero-layered structures in order to achieve a better performance in energy-related applications. In this theoretical work, we employ first-principles calculations to investigate a novel tetragonal (t-)ZnS/ZnSe hetero-bilayer (BL), as well as bilayers of ZnS and ZnSe only. A significant quantum confinement effect has been observed for all bilayers by state-of-the-art computations based upon the quasiparticle G0W0 approach and the Bethe-Salpeter equation (BSE). Among them, both t-ZnSe and t-ZnS/ZnSe BLs have strong optical absorption in the solar spectrum, rendering them potentially good candidates for solar energy harvesting. Moreover, the t-ZnS/ZnSe BL demonstrates good electron-hole separation by allocation of photo-induced electrons and holes in different layers, making it potentially suitable for applications in photocatalytic systems.


Patterns of exchange sex and HIV infection in high-risk heterosexual men and women.
Heterosexual partnerships involving the trade of money or goods for sex are a well-described HIV risk factor in Africa and Southeast Asia, but less research has been conducted on exchange partnerships and their impact on HIV infection in the United States. In our study, men and women were recruited from high-risk risk neighborhoods in New York City through respondent-driven sampling in 2006-2007. We examined the factors associated with having an exchange partner in the past year, the relationship between exchange partnerships and HIV infection, and the risk characteristics of those with exchange partners by the directionality of payment. Overall, 28% of men and 41% of women had a past-year exchange partner. For men, factors independently associated with exchange partnerships were older age, more total sexual partners, male partners, and frequent non-injection drug use. For women, factors were homelessness, more total sexual partners, more unprotected sex partners, and frequent non-injection drug use. Exchange partnerships were associated with HIV infection for both men and women, although the relationships were substantially confounded by other behavioral risks. Those who both bought and sold sex exhibited the highest levels of risk with their exchange and non-exchange partners. Exchange partnerships may be an HIV risk both directly and indirectly, given the overlap of this phenomenon with other risk factors that occur with both exchange and non-exchange partners.


Allergy to betalactam antibiotics in children: a prospective follow-up study in retreated children after negative responses in skin and challenge tests.
Up to 10% of the patients in whom suspected betalactam hypersensitivity (HS) has been excluded by skin and challenge tests report suspected allergic reactions during subsequent treatments with the same or very similar betalactams. It has been suggested that the reactions may result from a resensitization induced by the challenge performed at the time of the allergological work-up. However, most patients did not undergo a second allergological work-up, to determine if the reactions resulted from betalactam HS or not. We aimed to determine if children diagnosed nonallergic to betalactams have tolerated subsequent treatments with the initially suspected and/or other betalactams, and, in case of a reaction, if the reaction resulted from betalactam HS. We sent a questionnaire concerning the clinical history of their children to the parents of 256 children previously diagnosed nonallergic to betalactams. A second allergological work-up was performed in the children reporting suspected allergic reactions during subsequent treatments with the same and/or other betalactams. Skin tests were performed with the soluble form of the suspected (or very similar) betalactams and other betalactams from the same and other classes. Skin test responses were assessed at 15-20 min (immediate), 6-8 h (semi-late) and 48-72 h (late). Oral challenge (OC) was performed in children with negative skin tests, either at the hospital (immediate and accelerated reactions), or at home (delayed reactions). A response was obtained from 141 children (55.3%). Forty-eight (34%) of those children had not been treated with the betalactams for whom a diagnosis of allergy had been ruled out previously. Seven (7.5%) of the 93 children who had been treated again reported suspected allergic reactions. Skin tests and OC were performed in six of those children, and gave negative results in five children. In one child previously diagnosed nonallergic to amoxicillin associated with clavulanic acid, we diagnosed a delayed HS to clavulanic acid and a serum sickness-like disease to cefaclor. Thus, the frequency of reactions resulting from betalactam HS in children with negative skin and challenge tests is very low, and does not exceed 2.1% (2/93) if we consider that the child which refused a second allergological work-up is really allergic to betalactams. Our results in a very large number of children show that reactions presumed to result from betalactam HS are rare in children in whom the diagnosis of betalactam allergy has been ruled out previously. Moreover, they suggest that, as shown for the initial reactions, most of the reactions during subsequent treatments are rather a consequence of the infectious diseases for whom betalactams have been prescribed than a result of betalactam HS. Finally, they suggest that the risk of resensitization by OC is very low, and do not support the notion that skin testing should be repeated in children diagnosed nonallergic to betalactams.


The diffusion of Magnet hospital recognition.
Magnet recognition is promoted by many in the practice community as the gold standard of nursing care quality. The Magnet hospital population has exploded in recent years, with about 8% of U.S. general hospitals now recognized. The purpose of this study was to identify the characteristics that distinguish Magnet-recognized hospitals from other hospitals within the framework of diffusion theory. We conceptualize Magnet recognition as an organizational innovation and Magnet-recognized hospitals as adopters of the innovation. We hypothesize that adoption is associated with selected characteristics of hospitals and their markets. The study population consists of the 3,657 general hospitals in the United States in 2008 located in metropolitan or micropolitan areas. We used logistic regression analysis to estimate the association of Magnet recognition with organizational and market characteristics. Empirical results support hypotheses that adoption is positively associated with hospital complexity and specialization, as measured by teaching affiliation, and with hospital size, slack resources, and not-for-profit or public ownership (vs. for-profit). Adopters also are more likely to be located in markets that are experiencing population growth and are more likely to have competitor hospitals within the market that also have adopted Magnet status. A positive association of adoption with baccalaureate nursing school supply is contrary to the hypothesized relationship. Because of its rapid recent growth, consideration of Magnet program recognition should be on the strategic planning agenda of hospitals and hospital systems. Hospital administrators, particularly in smaller, for-profit hospitals, may expect more of their larger not-for-profit competitors, particularly teaching hospitals, to adopt Magnet recognition, increasing competition for baccalaureate-prepared registered nurses in the labor market.


Systems Biology Graphical Notation: Process Description language Level 1 Version 2.0.
The Systems Biology Graphical Notation (SBGN) is an international community effort that aims to standardise the visualisation of pathways and networks for readers with diverse scientific backgrounds as well as to support an efficient and accurate exchange of biological knowledge between disparate research communities, industry, and other players in systems biology. SBGN comprises the three languages Entity Relationship, Activity Flow, and Process Description (PD) to cover biological and biochemical systems at distinct levels of detail. PD is closest to metabolic and regulatory pathways found in biological literature and textbooks. Its well-defined semantics offer a superior precision in expressing biological knowledge. PD represents mechanistic and temporal dependencies of biological interactions and transformations as a graph. Its different types of nodes include entity pools (e.g. metabolites, proteins, genes and complexes) and processes (e.g. reactions, associations and influences). The edges describe relationships between the nodes (e.g. consumption, production, stimulation and inhibition). This document details Level 1 Version 2.0 of the PD specification, including several improvements, in particular: 1) the addition of the equivalence operator, subunit, and annotation glyphs, 2) modification to the usage of submaps, and 3) updates to clarify the use of various glyphs (i.e. multimer, empty set, and state variable).


The effect of zinc chloride, humidity and the substrate on the reaction of 1,2-indanedione-zinc with amino acids in latent fingermark secretions.
Anecdotal evidence from forensic practitioners and studies conducted under controlled conditions have indicated that the reaction between 1,2-indanedione and the amino acids present in latent fingermark deposits is highly susceptible to ambient humidity. The addition of catalytic amounts of zinc chloride to the 1,2-indanedione working solution--usually in the order of 1:25 to 1:4 molar ratio (indanedione:zinc)--significantly improves the colour and luminescence of fingermarks treated under dry conditions but appears to have a negligible effect on fingermarks treated in humid environments. The results presented in this paper confirmed that zinc(II) ions added to the 1,2-indanedione working solution act as a Lewis acid catalyst, stabilising a key intermediate during a rate-limiting hydrolysis step. Furthermore, studying the reaction using a chromatography-grade cellulose substrate method previously reported confirmed that cellulose substrates play a major role in facilitating the indanedione-amino acid reaction by acting as a surface catalyst in the early stages of the reaction and by directing the formation of the desired luminescent product (Joullié's Pink).


The Robert Wood Johnson Foundation Nurse Faculty Scholars Program: an opportunity for junior nurse faculty.
The Robert Wood Johnson Foundation Nurse Faculty Scholars program provides promising junior faculty extramural funding, expert mentoring, and the training needed to be successful in the academic role. The Nurse Faculty Scholars program, which admitted its first cohort in 2008, is designed to address the nursing faculty shortage by enhancing leadership, educational, and research skills in junior nursing faculty. This article provides an overview of the program, its purpose, and its eligibility requirements. The authors give strategies for selecting mentors, developing the written application, and preparing for an oral interview. Finally, the authors provide an analysis of funded institutions, research design and methods from current and recently funded projects, and rank and positions held by nursing mentors.


Accuracy of self-reports of Pap and mammography screening compared to medical record: a meta-analysis.
To conduct a systematic review and meta-analysis of the accuracy of self-reported Pap smear and mammography screening compared to medical record. About 37 articles were reviewed and accuracy indices of self-report were calculated. Meta-analysis with random effects was used. Study heterogeneity was investigated and meta-regressions were done including in the models those factors that were hypothesized, a priori, to potentially explain heterogeneity. Pooled sensitivity and specificity for Pap recall were 94.7% (95% confidence interval [CI]; 93.0%-96.4%) and 47.4% (95% CI; 39.0%-55.7%), and for mammography were 94.9% (95% CI; 93.4%-96.4%) and 61.8% (95% CI; 54.1%-69.5%), respectively. There was significant heterogeneity for all indices. Stratifying by the study population source (population versus clinic-based), population characteristics (minority or low socio-economic status versus not), length of recall (within past 12 months versus longer), and expected completeness of the medical record (authors searched radiology or pathology reports of all likely facilities women may have attended, versus studies that did not) did not eliminate heterogeneity. Women tend to over-report their participation in Pap and mammography screening in a given timeframe. The pooled estimates should be interpreted with caution due to unexplained heterogeneity.


Lack of oral health care for adults in harlem: a hidden crisis.
Profound and growing disparities exist in oral health among certain US populations. We sought here to determine the prevalence of oral health complaints among Harlem adults by measures of social class, as well as their access to oral health care. A population-based survey of adults in Central Harlem was conducted from 1992 to 1994. Two questions on oral health were included: whether participants had experienced problems with their teeth or gums during the past 12 months and, if so, whether they had seen a dentist. Of 50 health conditions queried about, problems with teeth or gums were the chief complaint among participants (30%). Those more likely to report oral health problems than other participants had annual household incomes of less than $9000 (36%), were unemployed (34%), and lacked health insurance (34%). The privately insured were almost twice as likely to have seen a dentist for oral health problems (87%) than were the uninsured (48%). There is an urgent need to provide oral health services for adults in Harlem. Integrating oral health into comprehensive primary care is one promising mechanism.


Prevalence of genetic susceptibility for breast and ovarian cancer in a non-cancer related study population: secondary germline findings from a Swiss single centre cohort.
Since the advent of high-throughput sequencing technologies, organised germline screening, independent of the personal and family cancer history, has been frequently proposed. Since ethnic and geographic populations significantly differ in their mutation spectra and prevalence, one critical prerequisite would be the knowledge of the expected carrier frequencies. For the first time, in a retrospective non-cancer related cohort from a single Swiss genetic centre, we systematically assessed the prevalence of secondary findings in 19 genes (BRCA1/2 plus 17 non-BRCA genes) previously designated by the US National Comprehensive Cancer Network (NCCN) for hereditary breast and ovarian cancer (HBOC) germline testing. A total of 400 individuals without a cancer diagnosis undergoing whole-exome sequencing (WES) analysis for neurodevelopmental disorders (NDDs) from 2015 to 2017 at IMG Zurich were included after quality assessment. Among these, 180 were unaffected parental couples, 27 unaffected parental singles and 13 NDD index patients (mean age 43 years). The majority of the cohort was of Caucasian ethnicity (n = 336, 84.0%) and of Northwest European ancestry (n = 202, 50.5%), for 70 of whom (42.5%) an autochthonous Swiss descent was assumed. For WES filtering of rare, potentially actionable secondary variants in HBOC genes, an overall minor allele frequency (MAF) below 0.65% was used as cut-off. Each rare variant was manually evaluated according to the recommended ACGM-AMP standards, with some adaptations including "hypomorphic" as an additional distinct pathogenicity class. Overall, 526 rare secondary variants (339 different variants) were encountered, with the BRCA1/2 genes accounting for 27.2% of the total variant yield. If stratified for variant pathogenicity, for BRCA1/2, three pathogenic variants were found in three females of Italian ancestry (carrier frequency of 0.8%). In the non-BRCA genes, five carriers of (likely) pathogenic variants (1.3%) were identified, with two Swiss individuals harbouring the same CHEK2 Arg160Gly variant known to be recurrent among Caucasians. Hence, the overall carrier rate added up to 2.0%. Additionally, seven various hypomorphic HBOC predisposing alleles were detected in 22 individuals (5.5%). We provide the first evidence of a high prevalence of HBOC-related cancer susceptibility in the heterogeneous Swiss general population and relevant subpopulations, particularly in individuals of Italian descent. These pioneering data may substantiate population-based HBOC screening in Switzerland.


Enkephalin analog prodrugs: assessment of in vitro conversion, enzyme cleavage characterization and blood-brain barrier permeability.
To improve the blood-brain barrier penetration of the delta-opioid receptor peptides [D-Pen2, D-Pen5]enkephalin (DPDPE) and [D-Pen2, L-Cys5]enkephalin (DPLCE), various prodrug forms were synthesized to increase lipophilicity and drug delivery to the brain. The aims of this study were 3-fold, 1) to assess the metabolic conversion of various DPDPE and DPLCE prodrugs in vitro using mouse brain homogenate and mouse serum, 2)to characterize the proteolytic enzymes responsible for cleaving prodrugs to the parent compounds using select peptidase inhibitors and 3)to assess the blood-brain barrier permeability of prodrugs, compared with their parent compounds, using the in vitro bovine brain microvessel endothelial cell culture model. The prodrugs with carboxyl-terminal phenylalanine residues (DPDPE-Phe and DPLCE-Phe) had significantly longer metabolic conversion times in both mouse serum and brain homogenates than did the prodrugs with amino-terminal phenylalanine residues. Inhibition of leucine aminopeptidase with bestatin in the serum increased the conversion time of Phe0-DPDPE from 6.8 min to 92.2 min. Inhibition of aminopeptidase M with amastatin in the brain homogenate increased the conversion time of Phe0-DPDPE from 3.9 min to > 450 min. The long half-life of DPLCE-Arg-Pro-Ala in serum (317 min) vs. brain (9.2 min) can be explained by the high levels of the degradative endopeptidase 24.15 (EC 126.96.36.199) in the central nervous system but not in plasma. The data also showed that, for specific prodrugs of DPDPE such as Phe0-DPDPE and DPDPE-Arg-Gly, the prodrug shows a significant improvement in permeability, compared with the parent compound. Therefore, these data provide evidence that prodrugs or prodrug-enzyme inhibitor combinations may optimize the delivery of peptide and/or protein drugs to the central nervous system.


Mesoscopic features of charge generation in organic semiconductors.
CONSPECTUS: In the past two decades, organic materials have been extensively investigated by numerous research groups worldwide for implementation in organic photovoltaic (OPV) devices. The interest in organic semiconductors is spurred by their potential low cost and facile tunability, making OPV devices a potentially disruptive technology. To study OPV operating mechanisms is also to explore a knowledge gap in our general understanding of materials, because both the time scales (femtosecond to microsecond) and length scales (nanometer to micrometer) relevant to OPV functionality occupy a challenging and fascinating space between the traditional regimes of quantum chemistry and solid-state physics. New theoretical frameworks and computational tools are needed to bridge the aforementioned length and time scales, and they must satisfy the criteria of computational tractability for systems involving 10(4)-10(6) atoms, while also maintaining predictive utility. While this challenge is far from solved, advances in density functional theory (DFT) have allowed researchers to investigate the ground- and excited-state properties of many intermediate sized systems (10(2)-10(3) atoms) that provide the outlines of the larger problem. Results on these smaller systems are already sufficient to predict optical gaps and trends in valence band energies, correct erroneous interpretations of experimental data, and develop models for charge generation and transport in OPV devices. The active films of high-efficiency OPV devices are comprised of mesoscopic mixtures of electron donor (D) and electron acceptor (A) species, a "bulk-heterojunction" (BHJ) device, subject to variable degrees of structural disorder. Depending on the degree of intermolecular electronic coupling and energy level alignment, the spatial delocalization of photoexcitations and charge carriers can affect the dynamics of the solar cell. In this Account, we provide an overview of three pivotal characteristics of solar cells that possess strong delocalization dependence: (1) the exciton binding energy, (2) charge transfer at the D-A heterojunction, and (3) the energy landscape in the vicinity of the D-A heterojunction. In each case, the length scale dependence can be assessed through DFT calculations on reference systems, with a view to establishing general trends. Throughout the discussion, we draw from the experimental and theoretical literature to provide a consistent view of what is known about these properties in actual BHJ blends. A consistent interpretation of the results to date affords the following view: transient delocalization effects and resonant charge transfer at the heterojunction are capable of funneling excitations away from trap states and mediating exciton dissociation; these factors alone are capable of explaining the remarkably good charge generation currently achieved in OPV devices. The exciton binding energy likely plays a minimal role in modern OPV devices, since the presence of the heterojunction serves to bypass the costly exciton-to-free-charge transition state.


The Neisseria meningitidis adhesion regulatory protein CrgA acts through oligomerization and interaction with RNA polymerase.
CrgA is a LysR-type transcriptional regulator involved in the intimate adhesion of Neisseria meningitidis to target human epithelial cells. It is negatively autoregulated, and its expression is transiently induced upon contact with target cells. We analysed the functional organization of CrgA using in frame deleted proteins. Four truncated proteins were constructed and purified. They were deleted between residues 20 and 40, 121 and 154, 111 and 181 and 268 and 291. Meningococcal mutants harbouring the corresponding deleted crgA alleles were also constructed. All mutants showed a reduced ability to adhere to epithelial cells. beta-Galactosidase activity assays using a crgA-lacZ transcriptional fusion showed that all the mutations except the 268-291 deletion resulted in loss of induction upon contact with target cells. Gel mobility shift assays and cross-linking assays showed that the oligomerization of CrgA is required for DNA binding and that the N-terminal part of CrgA is directly involved in DNA binding through a helix-turn-helix motif. The C-terminal region is also involved in DNA binding, probably by permitting the oligomerization of CrgA. The C-terminal region also seemed to interact with RNA polymerase. Therefore, the binding of CrgA and its interaction with RNA polymerase may inhibit the clearance of meningococcal promoters that are repressed by CrgA during the intimate adhesion of N. meningitidis to target cells.


Common features of protein unfolding and dissolution of hydrophobic compounds.
Protein unfolding and the dissolution of hydrophobic compounds (including solids, liquids, and gases) in water are characterized by a linear relation between entropy change and heat capacity change. The same slope is found for various classes of compounds, whereas the intercept depends on the particular class. The feature common to these processes is exposure of hydrophobic groups to water. These observations make possible the assignment of the heat capacity change to hydrophobic solvation and lead to the description of protein stability in terms of a hydrophobic and a nonhydrophobic contribution. A general representation of protein stability is given by the heat capacity change and the temperature.


The Science of Prognosis in Psychiatry: A Review.
Prognosis is a venerable component of medical knowledge introduced by Hippocrates (460-377 BC). This educational review presents a contemporary evidence-based approach for how to incorporate clinical risk prediction models in modern psychiatry. The article is organized around key methodological themes most relevant for the science of prognosis in psychiatry. Within each theme, the article highlights key challenges and makes pragmatic recommendations to improve scientific understanding of prognosis in psychiatry. The initial step to building clinical risk prediction models that can affect psychiatric care involves designing the model: preparation of the protocol and definition of the outcomes and of the statistical methods (theme 1). Further initial steps involve carefully selecting the predictors, preparing the data, and developing the model in these data. A subsequent step is the validation of the model to accurately test its generalizability (theme 2). The next consideration is that the accuracy of the clinical prediction model is affected by the incidence of the psychiatric condition under investigation (theme 3). Eventually, clinical prediction models need to be implemented in real-world clinical routine, and this is usually the most challenging step (theme 4). Advanced methods such as machine learning approaches can overcome some problems that undermine the previous steps (theme 5). The relevance of each of these themes to current clinical risk prediction modeling in psychiatry is discussed and recommendations are given. Together, these perspectives intend to contribute to an integrative, evidence-based science of prognosis in psychiatry. By focusing on the outcome of the individuals, rather than on the disease, clinical risk prediction modeling can become the cornerstone for a scientific and personalized psychiatry.


UK Role 4 military infectious diseases at Birmingham Heartlands Hospital in 2005-9.
Infectious diseases affecting British troops are mostly due to gastrointestinal and respiratory illnesses, but these are usually minor in severity, easy to manage and short in duration. To assess the importance of infections that are more severe, difficult to manage or longer in duration, it is necessary to look at military cases that are evacuated or otherwise referred to the UK Role 4 (definitive care) medical facility for infectious diseases. Case notes from military infectious disease patients seen at Birmingham Heartlands Hospital in 2005-2009 were reviewed to extract data on demographics, origin of infection, diagnostic categories, exact diagnoses, type and duration of care, time off duty, quality of care and costs incurred. Over a 4-year period, 138 cases were referred, 131 (95%) were male and 98 (71%) were from the Army. The origin of infection was Afghanistan in 52 (38%) and Belize in 19 (14%). From 131 patients (95%) that attended, 59 (45%) had dermatological illnesses and 38 (29%) had undifferentiated febrile illnesses. Diagnoses included 35 (27%) with cutaneous leishmaniasis and 21 (16%) with "Helmand Fever" due to sandfly fever, acute Q fever or rickettsial infection. For 51 in-patients, the median (range) length of stay was 3 (1-17) days and time off duty was 20 (5-127) days. For 80 out-patients, the median (range) number of attendances was 1 (1-23) and time off duty was 22 (1-228) days. All cases were seen promptly (within 7 days for in-patients and 28 days for out-patients), but only 59 (45%) had appropriate letters sent to the referring medical officer and none had F Med 85 notifications of infectious disease submitted. Aeromedical evacuation costs could not be calculated, but UK hospital care cost approximately pound 78 000 per year. Dermatological infections and undifferentiated febrile illnesses that require management at a UK Role 4 facility are important causes of disease non-battle injury. Prospective collection of this data in the future will be a valuable asset.


Monocyte trafficking in acute and chronic inflammation.
Environmental signals at the site of inflammation mediate rapid monocyte mobilization and dictate differentiation programs whereby these cells give rise to macrophages or dendritic cells. Monocytes participate in tissue healing, clearance of pathogens and dead cells, and initiation of adaptive immunity. However, recruited monocytes can also contribute to the pathogenesis of infection and chronic inflammatory disease, such as atherosclerosis. Here, we explore monocyte trafficking in the context of acute inflammation, relying predominantly on data from microbial infection models. These mechanisms will be compared to monocyte trafficking during chronic inflammation in experimental models of atherosclerosis. Recent developments suggest that monocyte trafficking shares common themes in diverse inflammatory diseases; however, important differences exist between monocyte migratory pathways in acute and chronic inflammation.


Comparison of inhibitory actions of prostacyclin and a new prostacyclin analogue on the aggregation of human platelet in whole blood.
Prostacyclin (PGI2), an unstable endogenous prostanoid, is a potent vasodilator and inhibitor of platelet aggregation. The use of exogenous PGI2 as an antithrombotic agent is limited by its chemical instability and lack of dissociation between its vasodilatory and antithrombotic actions. Iloprost (ZK36374) is a recently developed, chemically stable, carbacyclin analogue of PGI2. Preliminary evaluation studies have shown a significant dissociation between vasodilatory and antithrombotic actions of Iloprost. We have compared the effects of PGI2 and Iloprost on platelet aggregation in human whole blood. Platelet aggregation was induced by ADP (4 microM), adrenaline (Adr, 0.4 microM) and arachidonic acid (AA, 0.4 mM). Aggregation was quantified as a fall in the number of single platelets counted using the Clay Adams Ultra Flo 100 whole blood platelet counter. In the absence of any PGI2 or Iloprost, each aggregating agent induced up to an 80% fall in the number of single platelets counted. When blood was pre-incubated with PGI2 (1-6 nM) or Iloprost (1-6 nM), aggregation responses to all three aggregating agents were inhibited in a dose-dependent manner. Iloprost was equipotent to PGI2 against ADP-induced aggregation but was more potent than PGI2 against Adr- and AA-induced aggregation. It is concluded that Iloprost, as a chemically stable PGI2 analogue, may be superior to PGI2 as an antithrombotic agent.


Genetic and environmental contributions to retrospectively reported DSM-IV childhood attention deficit hyperactivity disorder.
A variety of methodologies and techniques converge on the notion that adults and children with attention deficit hyperactivity disorder (ADHD) have similar deficits, but there is limited knowledge about whether adult retrospective reports reflect similar genetic and environmental influences implicated in childhood ADHD. DSM-IV ADHD symptoms were collected retrospectively from 3896 young adults participating in the National Longitudinal Study of Adolescent Health. Responses from this genetically informative sample of same- and opposite-sex twins and siblings were used to determine the magnitude of genetic and environmental influences. Possible gender differences in these effects were also examined. The degree of familial specificity of the genetic and environmental influences on the Inattentive and Hyperactive-Impulsive symptom dimensions was also determined. Additive genetic effects contributed moderately to DSM-IV Inattentive, Hyperactive-Impulsive and Combined ADHD subtypes (heritability estimates of 0.30-0.38). Individual-specific influences accounted for the remaining proportion of the variance. Both genetic and individual-specific environmental effects contributed to the covariation of Inattentive and Hyperactive-Impulsive symptomologies. Results from our genetic analyses agree with previous findings based on self-assessment of current and retrospectively reported ADHD symptoms in adolescents and adults. Large individual-specific environmental influences as identified here suggest that current questionnaires used for retrospective diagnoses may not provide the most accurate reconstruction of the etiological influences on childhood ADHD in general population samples.


Long-term evaluation of quality of life and gastrointestinal well-being after segmental colo-rectal resection for deep infiltrating endometriosis (ENDO-RESECT QoL).
The primary objective is to assess the long-term quality of life (QoL) and gastrointestinal well-being in patients with endometriosis (DIE) who underwent segmental resection (SR), through specific questionnaires focused on endometriosis and specific gastrointestinal evaluation. The secondary objectives are represented by the evaluation of peri-operative and post-operative outcomes of the procedure. This observational cohort study ENDO-RESECT (ClinicalTrials.gov ID: NCT03824054) reports all clinical data about women who underwent SR for DIE between October 2005 and November 2017. In the part of the study dedicated to the QoL assessment, the questionnaires adopted were the Endometriosis Health Profile (EHP30), the Psychological General Well-Being Index and the Hospital Anxiety and Depression Scale, the Gastrointestinal Well-being questionnaire and the Bristol Stool chart. Major post-surgical morbidity and obstetric outcomes were also collected. 50 women (18% stage III and 82% stage IV rAFS) were considered for enrollment. EHP-30 interpretation demonstrated a significant improvement in all continuous variables, except for fertility concerns. The overall gastrointestinal QoL and most of the specific symptoms improved after surgery. Frequent bowel movements appeared in the 13% of the series not resulting in an impairment of general and gastrointestinal QoL. Constipation remained unchanged. Patients with depressive mood managed with laparoscopy, benefited the most from SR; moreover, patients with multinodular bowel localizations experienced a greater reduction in abdominal pain. Median FU after SR was 42.5 months (range 12-157 months). Only three (6%) cases of late major grade III complications were documented. The pregnancy rate was 50%. Improvement of general QoL and most of gastrointestinal symptoms was documented after SR.


Chemoprevention of mammary, cervix and nervous system carcinogenesis in animals using cultured Panax ginseng drugs and preliminary clinical trials in patients with precancerous lesions of the esophagus and endometrium.
The anticarcinogenic effects and mechanisms of the biotechnological drugs of Panax ginseng C.A. Meyer cultivated in Russia, bioginseng, panaxel and panaxel- 5, were studied. Bioginseng was produced from a tissue culture of ginseng root cultured on standard medium, whereas panaxel and panaxel-5 were produced from ginseng tissue root cultures using standard mediums enriched with 2-carboxyethylgermanium sesquioxide and 1-hydroxygermatran-monohydrate respectively. All three ginseng drugs inhibited the development of mammary tumors induced by intramammary injections of N-methyl-N-nitrosourea (MNU) in rats, the development of the brain and spinal cord tumors induced by transplacental administration of N-ethyl-N-nitrosourea (ENU) in rats, and the development of uterine, cervical and vaginal tumors induced by intravaginal applications of 7,12-dimethylbenz(a)anthracene (DMBA) in mice. The ginseng drugs induced the cytotoxic activity of macrophages in mice, enhanced T-lymphocyte rosette formation in guinea pigs exposed to cyclophosphamide, and stimulated the production of thyroid hormones in rats. These mechanisms may contribute to the anticarcinogenic action of the ginseng drugs. The organic germanium compounds present in panaxel and panaxel-5 did not potentiate the anticarcinogenic or immuno- stimulatory effects as much as biogeinseng. Preliminary clinical trials with panaxel and bioginseng were carried out in patients with precancerous lesions of the esophagus and endometrium. Panaxel was found to have a strong therapeutic effect in patients suffering from chronic erosive esophagitis. Bioginseng induced the regression of adenomatous-cystic hyperplasia of the endometrium in some patients. Thus, we conclude that the drugs of ginseng appear to hold considerable promise for future cancer chemoprevention.


Pharmacokinetic aspects of protriptyline plasma levels.
Plasma levels of protriptyline have been determined in 30 patients undergoing antidepressant therapy. After 3 1/2 weeks treatment at dosage levels of 40 mg/day, protriptyline plasma levels ranged from 430 to 1430 nmol/l. During this period only two-thirds of the subjects had definitely achieved asymptotic concentrations. Single dose studies in 5 volunteers suggest that the volume of distribution of protriptyline shows little intersubject variation. The half life of the drug, however, may vary appreciably from subject to subject, ranging from 54 to 198 h. The effects of two sedatives on mean protriptyline plasma levels have been determined. Mean plasma levels for nitrazepam recipients are indistinguishable from those for patients receiving no night sedation. The mean plasma levels for a group of patients receiving sodium amylobarbitone were significantly reduced. The problems of choice and early adjustment of dosages in order to achieve satisfactory plasma levels is discussed. For practical purposes it is suggested that early values may be of predictive significance in allowing early dosage adjustments to be made.


Aneurysm Morphology and Prediction of Rupture: An International Study of Unruptured Intracranial Aneurysms Analysis.
There are conflicting data between natural history studies suggesting a very low risk of rupture for small, unruptured intracranial aneurysms and retrospective studies that have identified a much higher frequency of small, ruptured aneurysms than expected. To use the prospective International Study of Unruptured Intracranial Aneurysms cohort to identify morphological characteristics predictive of unruptured intracranial aneurysm rupture. A case-control design was used to analyze morphological characteristics associated with aneurysm rupture in the International Study of Unruptured Intracranial Aneurysms database. Fifty-seven patients with ruptured aneurysms during follow-up were matched (by size and location) with 198 patients with unruptured intracranial aneurysms without rupture during follow-up. Twelve morphological metrics were measured from cerebral angiograms in a blinded fashion. Perpendicular height (P = .008) and size ratio (ratio of maximum diameter to the parent vessel diameter; P = .01) were predictors of aneurysm rupture on univariate analysis. Aspect ratio, daughter sacs, multiple lobes, aneurysm angle, neck diameter, parent vessel diameter, and calculated aneurysm volume were not statistically significant predictors of rupture. On multivariate analysis, perpendicular height was the only significant predictor of rupture (Chi-square 7.1, P-value .008). This study underscores the importance of other morphological factors, such as perpendicular height and size ratio, that may influence unruptured intracranial aneurysm rupture risk in addition to greatest diameter and anterior vs posterior location.


Induction of apoptosis by S-nitrosoglutathione and Cu2+ or Ni2+ ion through modulation of bax, bad, and bcl-2 proteins in human colon adenocarcinoma cells.
In this study, the amount of S-nitrosoglutathione (GSNO) was measured spectrophotometrically at 334 nm. A spontaneous decrease in absorbency at 334 nm was detected when GSNO was exposed to 37 degrees C and a high pH (pH 8.0). We investigated the catalytic roles of various metal ions on the decomposition of GSNO. The degradation of GSNO (0.5 mM) was enhanced by the presence of Cu(2+) and Ni(2+) ions. The amount of nitric oxide (NO) released from GSNO degradation was estimated by the Griess reaction based on nitrite accumulation. The results indicated that nitrite production was elevated at least twofold in the presence of Cu(2+). Our study further indicated that Cu(2+) enhanced GSNO-induced apoptosis in human colon adenocarcinoma HT 29 cells. We also found that copper ions modulated the expression of bad, bax, and bcl-2 in GSNO-treated HT 29 cells. The levels of bax and bad proteins in treated cells were significantly elevated about fourfold to sixfold when compared with mock-treated cells 24 h after combined treatment with GSNO plus Cu(2+) or Ni(2+). On the other hand, significant inhibition of bcl-2 occurred in HT 29 cells with simultaneous treatment of GSNO with Cu(2+) (or Ni(2+)). It seemed that Cu(2+) and Ni(2+) can enhance the decomposition of GSNO, which liberates NO to activate the pathways. Our results demonstrated that the apoptotic effects induced by GSNO were promoted by Ni(2+) and Cu(2+) through two different mechanisms: depletion of intracellular glutathione and triggering of NO release from GSNO, which then promotes NO-induced apoptosis in human cells.


Social learning solves the problem of narrow-peaked search landscapes: experimental evidence in humans.
The extensive use of social learning is considered a major reason for the ecological success of humans. Theoretical considerations, models and experiments have explored the evolutionary basis of social learning, showing the conditions under which learning from others is more adaptive than individual learning. Here we present an extension of a previous experimental set-up, in which individuals go on simulated 'hunts' and their success depends on the features of a 'virtual arrowhead' they design. Individuals can modify their arrowhead either by individual trial and error or by copying others. We study how, in a multimodal adaptive landscape, the smoothness of the peaks influences learning. We compare narrow peaks, in which solutions close to optima do not provide useful feedback to individuals, to wide peaks, where smooth landscapes allow an effective hill-climbing individual learning strategy. We show that individual learning is more difficult in narrow-peaked landscapes, but that social learners perform almost equally well in both narrow- and wide-peaked search spaces. There was a weak trend for more copying in the narrow than wide condition, although as in previous experiments social information was generally underutilized. Our results highlight the importance of tasks' design space when studying the adaptiveness of high-fidelity social learning.


The structure-sensitivity of memory access: evidence from Mandarin Chinese.
The present study examined the processing of the Mandarin Chinese long-distance reflexive ziji to evaluate the role that syntactic structure plays in the memory retrieval operations that support sentence comprehension. Using the multiple-response speed-accuracy tradeoff (MR-SAT) paradigm, we measured the speed with which comprehenders retrieve an antecedent for ziji. Our experimental materials contrasted sentences where ziji's antecedent was in the local clause with sentences where ziji's antecedent was in a distant clause. Time course results from MR-SAT suggest that ziji dependencies with syntactically distant antecedents are slower to process than syntactically local dependencies. To aid in interpreting the SAT data, we present a formal model of the antecedent retrieval process, and derive quantitative predictions about the time course of antecedent retrieval. The modeling results support the Local Search hypothesis: during syntactic retrieval, comprehenders initially limit memory search to the local syntactic domain. We argue that Local Search hypothesis has important implications for theories of locality effects in sentence comprehension. In particular, our results suggest that not all locality effects may be reduced to the effects of temporal decay and retrieval interference.


Thiol-Ene Alginate Hydrogels as Versatile Bioinks for Bioprinting.
Bioprinting is a powerful technique that allows precise and controlled 3D deposition of biomaterials in a predesigned, customizable, and reproducible manner. Cell-laden hydrogel ("bioink") bioprinting is especially advantageous for tissue engineering applications as multiple cells and biomaterial compositions can be selectively dispensed to create spatially well-defined architectures. Despite this promise, few hydrogel systems are easily available and suitable as bioinks, with even fewer systems allowing for molecular design of mechanical and biological properties. In this study, we report the development of a norbornene functionalized alginate system as a cell-laden bioink for extrusion-based bioprinting, with a rapid UV-induced thiol-ene cross-linking mechanism that avoids acrylate kinetic chain formation. The mechanical and swelling properties of the hydrogels are tunable by varying the concentration, length, and structure of dithiol PEG cross-linkers and can be further modified by postprinting secondary cross-linking with divalent ions such as calcium. The low concentrations of alginate needed (<2 wt %), coupled with their rapid in situ gelation, allow both the maintenance of high cell viability and the ability to fabricate large multilayer or multibioink constructs with identical bioprinting conditions. The modularity of this bioink platform design enables not only the rational design of materials properties but also the gel's biofunctionality (as shown via RGD attachment) for the expected tissue-engineering application. This modularity enables the creation of multizonal and multicellular constructs utilizing a chemically similar bioink platform. Such tailorable bioink platforms will enable increased complexity in 3D bioprinted constructs.


A fitness advantage for Aedes aegypti and the viruses it transmits when females feed only on human blood.
Literature on arthropod-borne diseases has traditionally supported the notion that mosquito vectors maintain a feeding duality that includes vertebrate blood meals for egg development and sugar meals from plants for the synthesis of flight and survival energy reserves. Aedes aegypti was found to deviate from that feeding pattern by obtaining a reproductive advantage when feeding only on human blood. Female mosquitoes fed human blood alone had a greater net replacement rate and intrinsic rate of growth during all phases of their reproductive life than conspecifics fed human blood plus sucrose. Feeding frequently on human hosts during each gonotrophic cycle is necessary to avoid death due to starvation and increases exponentially the spread of Ae. aegypti-borne disease. Our results help explain why Ae. aegypti is such an unusually efficient vector of human disease; frequent biting of humans results in a high reproductive rate for vectors as well as the viruses they transmit.


[Chromomycosis: report of a disabling case].
Chromomycosis is a deep subcutaneous mycosis caused by different dymorphic fungi species that normally live in vegetal debris. We report the case of a 51 year-old patient that six years previous to the evaluation worked making roof tiles in Madre de Dios, Peru; where he presented an initial papular lesion in a leg, which continued expanding until the 4 limbs were affected with disabling verrucous lesions. Fumagoid cells were found in the skin biopsy. The patient was hospitalized and received topical cleaning, antibiotics and terbinafine. He was discharged two months later with clinical improvement.


Molecular and clinical studies of Dent's disease in Japan: biochemical examination and renal ultrasonography do not predict carrier state.
Dent's disease is an X-linked renal tubular disorder characterized by low-molecular-weight-proteinuria, hypercalciuria, nephrolithiasis and renal failure. The disease is due to inactivation of a renal chloride channel gene, CLCN5. We have investigated 3 unrelated Japanese families for CLCN5 mutations and assessed the carrier mothers biochemically and ultrasonogaraphically to ascertain whether these clinical examinations can predict the carrier state of the disease. Twelve members from these families were studied biochemically and ultrasonographically. Leukocyte DNA from probands was used with CLCN5-specific primers for PCR amplification of the coding region and exon-intron boundaries, and the DNA sequences of the products determined to identify abnormalities in the gene. Three novel CLCN5 mutations consisting of a single base "A" insertion between nucleotides 590 and 591, a nonsense mutation (R28X) and a missense mutation (G506R) were exhibited. Hypophosphatemia was detected in 2 patients, beta2-microglobulinuria, alpha1-microglobulinuria, and hyperretinol binding proteinuria in 6 patients, hypercalciuria in 5 patients, decreased urine osmolality in 3 patients, and nephrocalcinosis or nephrolithiasis in 4 patients. Biochemical analysis of the urine and the renal ultrasonography in each carrier mother were completely normal. Neither urinary low-molecular-weight-proteins, urinary calcium to creatinine ratio, nor renal ultrasonography was predictive of carrier state in the 3 families with this disease, although each carrier mother had CLCN5 mutation. Hypophosphatemia and decreased urine osmolality might be a hint to suspect the carrier state of Dent's disease, although these findings are not found frequently.


Trends of breast cancer incidence in Iran during 2004-2008: a Bayesian space-time model.
Breast cancer is the most frequently diagnosed cancer in women and estimating its relative risks and trends of incidence at the area-level is helpful for health policy makers. However, traditional methods of estimation which do not take spatial heterogeneity into account suffer from drawbacks and their results may be misleading, as the estimated maps of incidence vary dramatically in neighboring areas. Spatial methods have been proposed to overcome drawbacks of traditional methods by including spatial sources of variation in the model to produce smoother maps. In this study we analyzed the breast cancer data in Iran during 2004-2008. We used a method proposed to cover spatial and temporal effects simultaneously and their interactions to study trends of breast cancer incidence in Iran. The results agree with previous studies but provide new information about two main issues regarding the trend of breast cancer in provinces of Iran. First, this model discovered provinces with high relative risks of breast cancer during the 5 years of the study. Second, new information was provided with respect to overall trend trends o. East-Azerbaijan, Golestan, North-Khorasan, and Khorasan-Razavi had the highest increases in rates of breast cancer incidence whilst Tehran, Isfahan, and Yazd had the highest incidence rates during 2004-2008. Using spatial methods can provide more accurate and detailed information about the incidence or prevalence of a disease. These models can specify provinces with different health priorities in terms of needs for therapy and drugs or demands for efficient education, screening, and preventive policy into action.


Contextualized Autonomy in Transitional Care for Youth With Neurologic Conditions: The Role of the Pediatric Neurologist.
Youth with neurologic conditions experience multiple life transitions. The transfer from pediatric to adult health care systems exemplifies one such complex and multifaceted transition that occurs in parallel with developmental, legal, and social changes that may influence the roles and responsibilities of youth and their caregivers. As a result, ethical situations, questions, and challenges may surface in transition care to which pediatric neurologists may be confronted. In this article, we focus on the topic of autonomy and situations that may arise in transition care in the context of pediatric neurology. Building from a clinical case, we present the concept of contextualized autonomy to work through the questions that arise in the case and propose ways of thinking through those challenging situations in transition care.


Adipsic hypernatremia without hypothalamic lesions accompanied by autoantibodies to subfornical organ.
Adipsic (or essential) hypernatremia is a rare hypernatremia caused by a deficiency in thirst regulation and vasopressin release. In 2010, we reported a case in which autoantibodies targeting the sensory circumventricular organs (sCVOs) caused adipsic hypernatremia without hypothalamic structural lesions demonstrable by magnetic resonance imaging (MRI); sCVOs include the subfornical organ (SFO) and organum vasculosum of the lamina terminalis (OVLT), which are centers for the monitoring of body-fluid conditions and the control of water and salt intakes, and harbor neurons innervating hypothalamic nuclei for vasopressin release. We herein report three newly identified patients (3- to 8-year-old girls on the first visit) with similar symptoms. The common features of the patients were extensive hypernatremia without any sensation of thirst and defects in vasopressin response to serum hypertonicity. Despite these features, we could not detect any hypothalamic structural lesions by MRI. Immunohistochemical analyses using the sera of the three patients revealed that antibodies specifically reactive to the mouse SFO were present in the sera of all cases; in one case, the antibodies also reacted with the mouse OVLT. The immunoglobulin (Ig) fraction of serum obtained from one patient was intravenously injected into wild-type mice to determine whether the mice developed similar symptoms. Mice injected with a patient's Ig showed abnormalities in water/salt intake, vasopressin release, and diuresis, which resultantly developed hypernatremia. Prominent cell death and infiltration of reactive microglia was observed in the SFO of these mice. Thus, autoimmune destruction of the SFO may be the cause of the adipsic hypernatremia. This study provides a possible explanation for the pathogenesis of adipsic hypernatremia without demonstrable hypothalamus-pituitary lesions.


The relationship between the haemoglobin concentration and the haematocrit in Plasmodium falciparum malaria.
Malaria is a very important cause of anaemia in tropical countries. Anaemia is assessed either by measurement of the haematocrit or the haemoglobin concentration. For comparisons across studies, it is often necessary to derive one measure from the other. Data on patients with slide-confirmed uncomplicated falciparum malaria were pooled from 85 antimalarial drug trials conducted in 25 different countries, to assess the haemoglobin/haematocrit relationship at different time points in malaria. Using a linear random effects model, a conversion equation for haematocrit was derived based on 3,254 measurements from various time points (ranging from day 0 to day 63) from 1,810 patients with simultaneous measurements of both parameters. Haemoglobin was also estimated from haematocrit with the commonly used threefold conversion. A good fit was obtained using Haematocrit = 5.62 + 2.60 * Haemoglobin. On average, haematocrit/3 levels were slightly higher than haemoglobin measurements with a mean difference (+/- SD) of -0.69 (+/- 1.3) for children under the age of 5 (n = 1,440 measurements from 449 patients). Based on this large data set, an accurate and robust conversion factor both in acute malaria and in convalescence was obtained. The commonly used threefold conversion is also valid.


Regionally distinct cutaneous afferent populations contribute to reflex modulation evoked by stimulation of the tibial nerve during walking.
During walking, cutaneous reflexes in ankle flexor muscle [tibialis anterior (TA)] evoked by tibial nerve (TIB) stimulation are predominantly facilitatory at early swing phase but reverse to suppression at late swing phase. Although the TIB innervates a large portion of the skin of the foot sole, the extent to which specific foot-sole regions contribute to the reflex reversals during walking remains unclear. Therefore, we investigated regional cutaneous contributions from discrete portions of the foot sole on reflex reversal in TA following TIB stimulation during walking. Summation effects on reflex amplitudes, when applying combined stimulation from foot-sole regions with TIB, were examined. Middle latency responses (MLRs; 70-120 ms) after TIB stimulation were strongly facilitated during the late stance to mid-swing phases and reversed to suppression just before heel (HL) strike. Both forefoot-medial (f-M) and forefoot-lateral stimulation in the foot sole induced facilitation during stance-to-swing transition phases, but HL stimulation evoked suppression during the late stance to the end of swing phases. At the stance-to-swing transition, a summation of MLR amplitude occurred only for combined f-M&TIB stimulation. However, the same was not true for the combined HL&TIB stimulation. At the swing-to-stance transition, there was a suppressive reflex summation only for HL&TIB stimulation. In contrast, this summation was not observed for the f-M&TIB stimulation. Our results suggest that reflex reversals evoked by TIB stimulation arise from distinct reflex pathways to TA produced by separate afferent populations innervating specific regions of the foot sole.


Influence of gastric acidity on the bioavailability of digoxin.
To study how changes in gastric acidity induced by omeprazole and pentagastrin affect the absorption of unchanged digoxin and its hydrolytic breakdown products. Double-blind, double-dummy, randomized, crossover study. Academic department of clinical pharmacology. Six healthy male volunteers. Subjects received digoxin, 1 mg orally, on three separate occasions: first, after pretreatment with omeprazole; second, after pretreatment with pentagastrin; and third, after "pretreatment" with placebo. The in-vitro decomposition of digoxin wa studied using a standard dissolution test. The urinary excretion of digoxin over a 120-hour period was measured using selective high-pressure liquid chromatography (HPLC) and a polarization enzyme immunoassay (EIA). Plasma concentrations were measured at 2 hours with the EIA. Digoxin was rapidly released from the tablets in the in-vitro test. At acid pH, decomposition (as measured with HPLC) was rapid. Pentagastrin reduced the urinary excretion of unchanged digoxin, as measured by HPLC, from 34% to 21.4% of the dose (difference, -12.6%; 95% Cl, -23.5 to -1.8; P less than 0.05), whereas omeprazole increased urinary excretion to 47.4% (difference, 13.4%; 95 Cl, 2.5% to 24.4%; P less than 0.05). However, such differences were not found with the nonselective polarization EIA. Our data suggest that gastric acidity causes the breakdown of digoxin to products that cross-react in the assay (EIA) that is commonly used clinically. In patients with reduced gastric acidity, increased plasma concentrations of unchanged digoxin may not be detected because of limitations of the EIA, which may invalidate the quantitative use of the plasma digoxin concentration as a predictor of digoxin toxicity. Omeprazole, and presumably other gastric-acid inhibitors, may increase the bioavailability of unchanged digoxin.


Folate-mediated one-carbon metabolism and neural tube defects: balancing genome synthesis and gene expression.
Neural tube defects (NTDs) refer to a cluster of neurodevelopmental conditions associated with failure of neural tube closure during embryonic development. Worldwide prevalence of NTDs ranges from approximately 0.5 to 60 per 10,000 births, with regional and population-specific variation in prevalence. Numerous environmental and genetic influences contribute to NTD etiology; accumulating evidence from population-based studies has demonstrated that folate status is a significant determinant of NTD risk. Folate-mediated one-carbon metabolism (OCM) is essential for de novo nucleotide biosynthesis, methionine biosynthesis, and cellular methylation reactions. Periconceptional maternal supplementation with folic acid can prevent occurrence of NTDs in the general population by up to 70%; currently several countries fortify their food supply with folic acid for the prevention of NTDs. Despite the unambiguous impact of folate status on NTD risk, the mechanism by which folic acid protects against NTDs remains unknown. Identification of the mechanism by which folate status affects neural tube closure will assist in developing more efficacious and better targeted preventative measures. In this review, we summarize current research on the relationship between folate status and NTDs, with an emphasis on linking genetic variation, folate nutriture, and specific metabolic and/or genomic pathways that intersect to determine NTD outcomes.


Glutamate receptor-induced 45Ca2+ accumulation in cortical cell culture correlates with subsequent neuronal degeneration.
Murine neuronal and glial cell cultures exposed briefly to glutamate accumulated large amounts of 45Ca2+ from the extracellular medium during the exposure. Most of the accumulation likely reflected influx into neurons, as little accumulation was observed in similarly treated glial cultures. When the concentration of glutamate was varied between 10 and 1000 microM, or exposure duration was varied between 0 and 10 min, the amount of 45Ca2+ accumulation correlated closely with the amount of neuronal death 24 hr later. Both 45Ca2+ accumulation and cell death could be attenuated in a dose-dependent manner by the competitive NMDA antagonist D-aminophosphonovalerate or the noncompetitive antagonist dextrorphan, with IC50 values of approximately 100 microM and 15 microM, respectively. In contrast, neither 45Ca2+ accumulation nor cell death was blocked by the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)/kainate antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) in the presence of high glycine. With brief exposure, high concentrations of AMPA, kainate, or K+ produced much less death or 45Ca2+ accumulation than produced by glutamate, especially if 10 microM MK-801 was included in the exposure medium to block NMDA receptor activation. Kainate- or AMPA-induced 45Ca2+ accumulation or neuronal cell death was blocked with CNQX. However, high K(+)-triggered 45Ca2+ accumulation was only partially blocked with CNQX plus MK-801, consistent with mediation by voltage-gated Ca2+ channels. In addition to measuring the accumulation of 45Ca2+ occurring during agonist exposure, we also assessed accumulation during the 30 min immediately following completion of a 3-5 min exposure to 500 microM NMDA.(ABSTRACT TRUNCATED AT 250 WORDS)


Quantification of plant chlorophyll content using Google Glass.
Measuring plant chlorophyll concentration is a well-known and commonly used method in agriculture and environmental applications for monitoring plant health, which also correlates with many other plant parameters including, e.g., carotenoids, nitrogen, maximum green fluorescence, etc. Direct chlorophyll measurement using chemical extraction is destructive, complex and time-consuming, which has led to the development of mobile optical readers, providing non-destructive but at the same time relatively expensive tools for evaluation of plant chlorophyll levels. Here we demonstrate accurate measurement of chlorophyll concentration in plant leaves using Google Glass and a custom-developed software application together with a cost-effective leaf holder and multi-spectral illuminator device. Two images, taken using Google Glass, of a leaf placed in our portable illuminator device under red and white (i.e., broadband) light-emitting-diode (LED) illumination are uploaded to our servers for remote digital processing and chlorophyll quantification, with results returned to the user in less than 10 seconds. Intensity measurements extracted from the uploaded images are mapped against gold-standard colorimetric measurements made through a commercially available reader to generate calibration curves for plant leaf chlorophyll concentration. Using five plant species to calibrate our system, we demonstrate that our approach can accurately and rapidly estimate chlorophyll concentration of fifteen different plant species under both indoor and outdoor lighting conditions. This Google Glass based chlorophyll measurement platform can display the results in spatiotemporal and tabular forms and would be highly useful for monitoring of plant health in environmental and agriculture related applications, including e.g., urban plant monitoring, indirect measurements of the effects of climate change, and as an early indicator for water, soil, and air quality degradation.


Direct and indirect quantification of mitral regurgitation with cardiovascular magnetic resonance, and the effect of heart rate variability.
Quantifying mitral regurgitation with cardiovascular magnetic resonance (CMR) involves indirect calculation, which increases the potential for error. We examined a direct quantification method using velocity mapping across the mitral valve, which may be less susceptible to error, and also examined the effect of heart rate variability on both techniques. Fifty-five patients underwent mitral regurgitation quantification with CMR by the direct method and two indirect methods-the standard method subtracting aortic flow (assessed by velocity mapping) from left ventricular stroke volume (assessed by cine imaging) and the 'volumetric' method using the difference between left and right ventricular stroke volumes. The methods were compared using Bland-Altman analyses. Patients with low heart rate variability (beat-to-beat variability <30 bpm; n = 44) showed good agreement between direct and indirect methods (95% confidence limits for the difference between measurements +/-16.7 ml/11.8% regurgitant fraction for the standard method; +/-21.7 ml/15.4% for the volumetric method), with no significant offset (mean difference +2.8 ml/+1.9% for standard and +3.1 ml/+2.3% for volumetric methods). Patients with high heart rate variability (>30 bpm; n = 11) showed poor agreement between techniques (95% limits +/-80.3 ml/56.0%) and significant offset (mean difference +31.7 ml/+19.5%). Direct quantification of mitral regurgitation with CMR compares well with indirect methods for patients with low heart rate variability, involves fewer calculations and is quick. All CMR measurements that use velocity mapping may be inaccurate, however, in patients with highly irregular rhythms and should be avoided in these patients.


Dehydration of bacterial cellulose and the water content effects on its viscoelastic and electrochemical properties.
Bacterial cellulose (BC) has interesting properties including high crystallinity, tensile strength, degree of polymerisation, water holding capacity (98%) and an overall attractive 3D nanofibrillar structure. The mechanical and electrochemical properties can be tailored upon incomplete BC dehydration. Under different water contents (100, 80 and 50%), the rheology and electrochemistry of BC were evaluated, showing a progressive stiffening and increasing resistance with lower capacitance after partial dehydration. BC water loss was mathematically modelled for predicting its water content and for understanding the structural changes of post-dried BC. The dehydration of the samples was determined via water evaporation at 37 °C for different diameters and thicknesses. The gradual water evaporation observed was well-described by the model proposed (R2 up to 0.99). The mathematical model for BC water loss may allow the optimisation of these properties for an intended application and may be extendable for other conditions and purposes.


Detection of cytochrome C in a single cell using an optical nanobiosensor.
In this work, the intracellular measurement of cytochrome c using an optical nanobiosensor is demonstrated. The nanobiosensor is a unique fiberoptics-based tool which allows the minimally invasive analysis of intracellular components. Cytochrome c is a very important protein to the process which produces cellular energy. In addition, cytochrome c is well-known as the protein involved in apoptosis, or programmed cell death. delta-Aminolevulinic acid (5-ALA) was used to induce apoptosis in MCF-7 human breast carcinoma cells. 5-ALA, a photodynamic therapy (PDT) drug in cells was activated by a HeNe laser beam. After the PDT photoactivation, the release of cytochrome c from the mitochondria to the cytoplasm in a MCF-7 cell was monitored by the optical nanobiosensor inserted inside the single cell and followed by an enzyme-linked immunosorbent assay (ELISA) outside the cell. The combination of the nanobiosensor with the ELISA immunoassay improved the detection sensitivity of the nanobiosensor due to enzymatic amplification. Our results lead to the investigation of an apoptotic pathway at the single cell level.


Quantitative evaluation of automated skull-stripping methods applied to contemporary and legacy images: effects of diagnosis, bias correction, and slice location.
Performance of automated methods to isolate brain from nonbrain tissues in magnetic resonance (MR) structural images may be influenced by MR signal inhomogeneities, type of MR image set, regional anatomy, and age and diagnosis of subjects studied. The present study compared the performance of four methods: Brain Extraction Tool (BET; Smith [2002]: Hum Brain Mapp 17:143-155); 3dIntracranial (Ward [1999] Milwaukee: Biophysics Research Institute, Medical College of Wisconsin; in AFNI); a Hybrid Watershed algorithm (HWA, Segonne et al. [2004] Neuroimage 22:1060-1075; in FreeSurfer); and Brain Surface Extractor (BSE, Sandor and Leahy [1997] IEEE Trans Med Imag 16:41-54; Shattuck et al. [2001] Neuroimage 13:856-876) to manually stripped images. The methods were applied to uncorrected and bias-corrected datasets; Legacy and Contemporary T1-weighted image sets; and four diagnostic groups (depressed, Alzheimer's, young and elderly control). To provide a criterion for outcome assessment, two experts manually stripped six sagittal sections for each dataset in locations where brain and nonbrain tissue are difficult to distinguish. Methods were compared on Jaccard similarity coefficients, Hausdorff distances, and an Expectation-Maximization algorithm. Methods tended to perform better on contemporary datasets; bias correction did not significantly improve method performance. Mesial sections were most difficult for all methods. Although AD image sets were most difficult to strip, HWA and BSE were more robust across diagnostic groups compared with 3dIntracranial and BET. With respect to specificity, BSE tended to perform best across all groups, whereas HWA was more sensitive than other methods. The results of this study may direct users towards a method appropriate to their T1-weighted datasets and improve the efficiency of processing for large, multisite neuroimaging studies.


Production and diversity of volatile terpenes from plants on calcareous and siliceous soils: effect of soil nutrients.
Fertilizer effects on terpene production have been noted in numerous reports. In contrast, only a few studies have studied the response of leaf terpene content to naturally different soil fertility levels. Terpene content, as determined by gas chromatography/mass spectrometry/flame ionization detector, and growth of Pinus halepensis, Rosmarinus officinalis, and Cistus albidus were studied on calcareous and siliceous soils under field conditions. The effect of nitrogen (N) and extractable phosphorus (P(E)) from these soils on terpenes was also investigated since calcareous soils mainly differ from siliceous soils in their higher nutrient loadings. Rich terpene mixtures were detected. Twenty-one terpenes appeared in leaf extracts of R. officinalis and C. albidus and 20 in P. halepensis. Growth of all species was enhanced on calcareous soils, while terpene content showed a species-specific response to soil type. The total monoterpene content of P. halepensis and that of some major compounds (e.g., delta-terpinene) were higher on calcareous than on siliceous soils. A significant and positive relationship was found between concentration of N and P(E) and leaf terpene content of this species. These findings suggest that P. halepensis may respond to an environment characterized by increasing soil deposition, by allocating carbon resources to the synthesis of terpene defense metabolites without growth reduction. Results obtained for R. officinalis showed high concentrations of numerous major monoterpenes (e.g., myrcene, camphor) in plants growing on calcareous soils, while alpha-pinene, beta-caryophyllene, and the total sesquiterpene content were higher on siliceous soils. Finally, only alloaromadendrene and delta-cadinene of C. albidus showed higher concentrations on siliceous soils. Unlike P. halepensis, soil nutrients were not involved in terpene variation in calcareous and siliceous soils of these two shrub species. Possible ecological explanations on the effect of soil type for these latter two species as well as the ecological explanation of rich terpene mixtures are discussed.


C-peptide is the appropriate outcome measure for type 1 diabetes clinical trials to preserve beta-cell function: report of an ADA workshop, 21-22 October 2001.
The underlying cause of type 1 diabetes, loss of beta-cell function, has become the therapeutic target for a number of interventions in patients with type 1 diabetes. Even though insulin therapies continue to improve, it remains difficult to achieve normal glycemic control in type 1 diabetes, especially long term. The associated risks of hypoglycemia and end-organ diabetic complications remain. Retention of beta-cell function in patients with type 1 diabetes is known to result in improved glycemic control and reduced hypoglycemia, retinopathy, and nephropathy. To facilitate the development of therapies aimed at altering the type 1 diabetes disease process, an American Diabetes Association workshop was convened to identify appropriate efficacy outcome measures in type 1 diabetes clinical trials. The following consensus emerged: While measurements of immune responses to islet cells are important in elucidating pathogenesis, none of these measures have directly correlated with the decline in endogenous insulin secretion. HbA(1c) is a highly valuable clinical measure of glycemic control, but it is an insensitive measure of beta-cell function, particularly with the currently accepted standard of near-normal glycemic control. Rates of severe hypoglycemia and diabetic complications ultimately will be improved by therapies that are effective at preserving beta-cell function but as primary outcomes require inordinately large and protracted trials. Endogenous insulin secretion is assessed best by measurement of C-peptide, which is cosecreted with insulin in a one-to-one molar ratio but unlike insulin experiences little first pass clearance by the liver. Measurement of C-peptide under standardized conditions provides a sensitive, well accepted, and clinically validated assessment of beta-cell function. C-peptide measurement is the most suitable primary outcome for clinical trials of therapies aimed at preserving or improving endogenous insulin secretion in type 1 diabetes patients. Available data demonstrate that even relatively modest treatment effects on C-peptide will result in clinically meaningful benefits. The development of therapies for addressing this important unmet clinical need will be facilitated by trials that are carefully designed with beta-cell function as determined by C-peptide measurement as the primary efficacy outcome.


Transcriptional profiling of rhesus monkey embryonic stem cells.
Embryonic stem cells (ESCs) may be able to cure or alleviate the symptoms of various degenerative diseases. However, unresolved issues regarding survival, functionality, and tumor formation mean a prudent approach should be adopted towards advancing ESCs into human clinical trials. The rhesus monkey provides an ideal model organism for developing strategies to prevent immune rejection and test the feasibility, safety, and efficacy of ESC-based medical treatments. Transcriptional profiling of rhesus monkey ESCs provides a foundation for pre-clinical ESC research in this species. In the present study, we used microarray technology, immunocytochemistry, reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time PCR (qPCR) to characterize and transcriptionally profile rhesus monkey ESCs. We identified 367 stemness gene candidates that were highly (>85%) conserved across five different ESC lines. Rhesus monkey ESC lines maintained a pluripotent undifferentiated state over a wide range of POU5F1 (also known as OCT4) expression levels, and comparisons between rhesus monkey, mouse, and human stemness genes revealed five mammalian stemness genes: CCNB1, GDF3, LEFTB, POU5F1, and NANOG. These five mammalian genes are strongly expressed in rhesus monkey, mouse, and human ESCs, albeit only in the undifferentiated state, and represent the core key mammalian stemness factors.


Fish predation by semi-aquatic spiders: a global pattern.
More than 80 incidences of fish predation by semi-aquatic spiders--observed at the fringes of shallow freshwater streams, rivers, lakes, ponds, swamps, and fens--are reviewed. We provide evidence that fish predation by semi-aquatic spiders is geographically widespread, occurring on all continents except Antarctica. Fish predation by spiders appears to be more common in warmer areas between 40° S and 40° N. The fish captured by spiders, usually ranging from 2-6 cm in length, are among the most common fish taxa occurring in their respective geographic area (e.g., mosquitofish [Gambusia spp.] in the southeastern USA, fish of the order Characiformes in the Neotropics, killifish [Aphyosemion spp.] in Central and West Africa, as well as Australian native fish of the genera Galaxias, Melanotaenia, and Pseudomugil). Naturally occurring fish predation has been witnessed in more than a dozen spider species from the superfamily Lycosoidea (families Pisauridae, Trechaleidae, and Lycosidae), in two species of the superfamily Ctenoidea (family Ctenidae), and in one species of the superfamily Corinnoidea (family Liocranidae). The majority of reports on fish predation by spiders referred to pisaurid spiders of the genera Dolomedes and Nilus (>75% of observed incidences). There is laboratory evidence that spiders from several more families (e.g., the water spider Argyroneta aquatica [Cybaeidae], the intertidal spider Desis marina [Desidae], and the 'swimming' huntsman spider Heteropoda natans [Sparassidae]) predate fish as well. Our finding of such a large diversity of spider families being engaged in fish predation is novel. Semi-aquatic spiders captured fish whose body length exceeded the spiders' body length (the captured fish being, on average, 2.2 times as long as the spiders). Evidence suggests that fish prey might be an occasional prey item of substantial nutritional importance.


A substrate sensor chip to assay the enzymatic activity of Botulinum neurotoxin A.
Botulinum neurotoxin A (BoNT/A) induces muscle paralysis by enzymatically cleaving the presynaptic SNARE protein SNAP-25, which results in lasting inhibition of acetylcholine release at the neuromuscular junction. A rapid and sensitive in vitro assay for BoNT/A is required to replace the mouse lethality assay (LD50) in current use. We have developed a fully automated sensor to assay the endoprotease activity of BoNT/A. We produced monoclonal antibodies (mAbs) that recognize SNAP-25 neo-epitopes specifically generated by BoNT/A action. Recombinant SNAP-25 was coupled to the sensor surface of a surface plasmon resonance (SPR) system and samples containing BoNT/A were injected over the substrate sensor. Online substrate cleavage was monitored by measuring binding of mAb10F12 to a SNAP-25 neo-epitope. The SNAP-25-chip assay was toxin serotype-specific and detected 55 fM BoNT/A (1 LD50/ml) in 5 min and 0.4 fM (0.01 LD50/ml) in 5h. Time-course and dose-response curves were linear, yielding a limit of quantification of 0.03 LD50/ml. This label-free method is 100 times more sensitive than the mouse assay, potentially providing rapid read-out of small amounts of toxin for environmental surveillance and the quality control of pharmaceutical preparations.