Evidence for a hopping mechanism in metal|single molecule|metal junctions involving conjugated metal-terpyridyl complexes; potential-dependent conductances of complexes [M(pyterpy)2]2+ (M = Co and Fe; pyterpy = 4'-(pyridin-4-yl)-2,2':6',2''-terpyridine) in ionic liquid.
Extensive studies of various families of conjugated molecules in metal|molecule|metal junctions suggest that the mechanism of conductance is usually tunnelling for molecular lengths < ca. 4 nm, and that for longer molecules, coherence is lost as a hopping element becomes more significant. In this work we present evidence that, for a family of conjugated, redox-active metal complexes, hopping may be a significant factor for even the shortest molecule studied (ca. 1 nm between contact atoms). The length dependence of conductance for two series of such complexes which differ essentially in the number of conjugated 1,4-C6H4- rings in the structures has been studied, and it is found that the junction conductances vary linearly with molecular length, consistent with a hopping mechanism, whereas there is significant deviation from linearity in plots of log(conductance) vs. length that would be characteristic of tunnelling, and the slopes of the log(conductance)-length plots are much smaller than expected for an oligophenyl system. Moreover, the conductances of molecular junctions involving the redox-active molecules, [M(pyterpy)2]2+/3+ (M = Co, Fe) have been studied as a function of electrochemical potential in ionic liquid electrolyte, and the conductance-overpotential relationship is found to fit well with the Kuznetsov-Ulstrup relationship, which is essentially a hopping description.


The endocannabinoid system and migraine.
The recently discovered endocannabinoid system (ECS), which includes endocannabinoids and the proteins that metabolize and bind them, has been implicated in multiple regulatory functions both in health and disease. Several studies have suggested that ECS is centrally and peripherally involved in the processing of pain signals. This finding is corroborated by the evidence that endocannabinoids inhibit, through a cannabinoid type-1 receptor (CB1R)-dependent retrograde mechanism, the release of neurotransmitters controlling nociceptive inputs and that the levels of these lipids are high in those regions (such as sensory terminals, skin, dorsal root ganglia) known to be involved in transmission and modulation of pain signals. In this review we shall describe experimental and clinical data that, intriguingly, demonstrate the link between endocannabinoids and migraine, a neurovascular disorder characterized by recurrent episodic headaches and caused by abnormal processing of sensory information due to peripheral and/or central sensitization. Although the exact ECS-dependent mechanisms underlying migraine are not fully understood, the available results strongly suggest that activation of ECS could represent a promising therapeutical tool for reducing both the physiological and inflammatory components of pain that are likely involved in migraine attacks.


Economic evaluation of voriconazole compared with conventional amphotericin B for the primary treatment of aspergillosis in immunocompromised patients.
The objective of this study was to conduct an economic evaluation of voriconazole compared with conventional amphotericin B deoxycholate (CAB) using data from a recently reported randomized comparative trial in patients with various underlying immunosuppressive conditions. This trial demonstrated the superiority of voriconazole in terms of clinical response, survival and safety when used as primary therapy for invasive aspergillosis. A decision analytic model was designed using an expert panel and populated primarily with efficacy and resource utilization data collected prospectively during the clinical trial. The analysis was carried out from the perspective of the health care system and all costs are reported in 2002 US dollars. Average total treatment costs per patient were 10% lower in the voriconazole arm ($30 664) than in the CAB arm ($34 144), resulting from reduced consumption of hospital resources and fewer changes in antifungal therapy. In the base case analysis, voriconazole provided an average saving of $3481 per treated patient, resulted in a lower cost per survivor ($43 310 versus $58 971) and a lower cost per successfully treated patient ($58 100 versus $108 124) compared with CAB. Sensitivity analyses demonstrated that the cost savings observed were maintained over a wide range of alternative values for both unit costs and resource utilization, including length of hospital stay, time spent in intensive care units, bed day costs and the cost of lipid formulations of amphotericin B. Incremental cost-effectiveness analysis indicated the dominance of voriconazole because of both lower costs and greater efficacy.


A substudy protocol of the hypertension in the Very Elderly Trial assessing cognitive decline and dementia incidence (HYVET-COG) : An ongoing randomised, double-blind, placebo-controlled trial.
Randomised, controlled trials and population studies have suggested a link between hypertension and the development of dementia (vascular dementia and Alzheimer's disease) although the results are not conclusive. The very elderly are at highest risk for both hypertension and dementia but have been underrepresented in studies to date. The Hypertension in the Very Elderly Trial (HYVET), an international, randomised, double-blind, placebo-controlled trial of antihypertensive medication for those > or =80 years of age, is currently underway. The protocol for the assessment of cognitive decline and detection of incident dementia cases is described in this article. The objective of this protocol is to determine whether treatment of hypertension in this very elderly group affects the incidence of dementia and decline in cognitive function during the period of the HYVET trial. Cognitive function in all HYVET trial participants is assessed at baseline and annually thereafter using the Mini-Mental State Examination (MMSE). An MMSE score that decreases more than three points per year or decreases to <24 prompts further investigation for possible incident dementia. This includes use of the Diagnostic and Statistical Manual-IV criteria, the modified Hachinski scale and CT scanning. In cases where a CT scan cannot be obtained the full Hachinski scale is used. Additional information is provided when available from the clock drawing test and Geriatric Depression Scale. Data are also gathered from all trial participants with regard to education, blood pressure, alcohol use, co-morbidities and concomitant treatments. At the end of the trial, changes in cognitive function and incident dementia cases will be compared in those receiving active treatment (indapamide slow-release +/- perindopril) and those receiving matching placebo. The protocol has been applied successfully and the data collection phase is underway. Previous population studies have been unable to study antihypertensive use in a controlled manner and randomised, controlled trials suggest a positive or mixed picture regarding an association between use of antihypertensive therapy and reduction in incident cognitive decline and/or dementia. No such trial has yet been performed exclusively in subjects > or =80 years of age with hypertension. The HYVET trial is placebo controlled and while investigating the cardiovascular effect of treatment also assesses cognitive function and quality of life in both the placebo and actively treated arms of the trial. The HYVET cognitive substudy trial (HYVET-COG) design has been successfully applied and the instruments chosen appear to be appropriate, valid and pragmatic. HYVET-COG provides the first opportunity to examine the effect of antihypertensive treatment on incident dementia in a high-risk very elderly population.


Application of finite element analysis to the design of tissue leaflets for a percutaneous aortic valve.
Percutaneous Aortic Valve (PAV) replacement is an attractive alternative to open heart surgery, especially for patients considered to be poor surgical candidates. Despite this, PAV replacement still has its limitations and associated risks. Bioprosthetic heart valves still have poor long-term durability due to calcification and mechanical failure. In addition, the implantation procedure often presents novel challenges, including damage to the expandable stents and bioprosthetic leaflets. In this study, a simplified version of Fung's elastic constitutive model for skin, developed by Sun and Sacks, was implemented using finite element analysis (FEA) and applied to the modelling of bovine and kangaroo pericardium. The FEA implementation was validated by simulating biaxial tests and by comparing the results with experimental data. Concepts for different PAV geometries were developed by incorporating valve design and performance parameters, along with stent constraints. The influence of effects such as different leaflet material, material orientation and abnormal valve dilation on the valve function was investigated. The stress distribution across the valve leaflet was also examined to determine the appropriate fibre direction for the leaflet. The simulated attachment forces were compared with suture tearing tests performed on the pericardium to evaluate suture density. It is concluded that kangaroo pericardium is suitable for PAV applications, and superior to bovine pericardium, due to its lower thickness and greater extensibility.


Evolutionarily conserved human targets of adenosine to inosine RNA editing.
A-to-I RNA editing by ADARs is a post-transcriptional mechanism for expanding the proteomic repertoire. Genetic recoding by editing was so far observed for only a few mammalian RNAs that are predominantly expressed in nervous tissues. However, as these editing targets fail to explain the broad and severe phenotypes of ADAR1 knockout mice, additional targets for editing by ADARs were always expected. Using comparative genomics and expressed sequence analysis, we identified and experimentally verified four additional candidate human substrates for ADAR-mediated editing: FLNA, BLCAP, CYFIP2 and IGFBP7. Additionally, editing of three of these substrates was verified in the mouse while two of them were validated in chicken. Interestingly, none of these substrates encodes a receptor protein but two of them are strongly expressed in the CNS and seem important for proper nervous system function. The editing pattern observed suggests that some of the affected proteins might have altered physiological properties leaving the possibility that they can be related to the phenotypes of ADAR1 knockout mice.


Induction of apoptosis by HIV-1-infected monocytic cells.
We have previously described a soluble 6000-Da peptide produced by an HIV-1-infected human macrophage cell line, clone 43(HIV), which induces apoptosis in T and B cells. We have identified this factor as the novel cDNA clone FL14676485 that encodes for the human hypothetical protein, FLJ21908. The FL14676485 cDNA clone was isolated from a 43(HIV) lambda ZAP Escherichia coli expression library and screened with a panel of rabbit and mouse anti-apoptotic Abs. We transfected the FL14676485 clone into Bosc cells and non-HIV-1-infected 43 cells. Western blot analysis of lysates from the FL14676485-transfected 43 cells and Bosc cells using anti-proapoptotic factor Abs revealed a protein with a molecular mass of 66 kDa corresponding to the size of the full-length gene product of the FL14676485 clone, while Western blot of the supernatant demonstrated a doublet of 46-kDa and 6000-Da peptide that corresponds to our previously described proapoptotic factor. Primary HIV-1(BaL)-infected monocytes also produce the FLJ21908 protein. Supernatants from these transfected cells induced apoptosis in PBMC, CD4(+), and CD8(+) T and B cells similar to the activity of our previously described proapoptotic factor. PCR analysis of 43 cells and 43(HIV) cells revealed a base pair fragment of 420 bp corresponding to the FL14676485 gene product in 43(HIV) cells, but not in 43 cells. The FLJ21908 protein induces apoptosis through activation of caspase-9 and caspase-3. We have further demonstrated that the FLJ21908 protein has apoptotic activity in the SH-SY5Y neuronal cell line and can be detected in brain and lymph tissue from HIV-1-infected patients who have AIDS dementia. The FLJ21908 protein may contribute to the apoptosis and dementia observed in AIDS patients.


Genetic variants involved in innate immunity modulate the risk of inflammatory bowel diseases in an understudied Malaysian population.
Inflammatory bowel diseases (IBD) are chronic gastrointestinal inflammatory conditions comprising two major subtypes: Crohn's disease (CD) and ulcerative colitis (UC). The incidence of IBD is increasing in Asian countries including Malaysia. The aim of this study was to determine whether 32 single nucleotide polymorphisms (SNPs) strongly associated with IBD from genome-wide association studies, performed mainly in Caucasian populations, are associated with IBD in a Malaysian population, correlating these findings with local and systemic inflammation. Selected SNPs were investigated in a Malaysian cohort comprising 36 IBD patients and 75 controls using customized matrix-assisted laser desorption ionization time-of-flight genotyping. Local mRNA and/or systemic protein levels of IL-10, IL-12, IL-22, IL-23, and TNF-α were measured in these same subjects. ATG16L2 rs11235667 and LINC00824 rs6651252 was significantly associated with increased CD risk while IL12B rs56167332 was a significant protective factor. Three SNPs (SBNO2 rs2024092, CARD9 rs10781499, and rs17085007 between GPR12-USP12) were significantly associated with increased UC risk while NKX2-3 rs4409764 was a significant protective factor. After adjusting for age, gender, and ethnicity, SBNO2 rs2024092, ATG16L2 rs11235667, CARD9 rs10781499, and LINC00824 rs6651252 remained associated with IBD. Interestingly, the risk alleles of IL10 rs3024505, CARD9 rs1078149, and IL12 rs6556412 were associated with higher levels of IL-10, IL-22, and IL-23 in these same subjects, respectively. This study identified eight SNPs associated with IBD and/or its subtypes in the Malaysia population, significantly advancing our understanding of the genetic contribution to IBD in this understudied population. Three of these SNPs modulated relevant cytokine levels and thus, may directly contribute to IBD pathogenesis.


Complete Genome Sequence of Serratia quinivorans Strain 124R, a Facultative Anaerobe Isolated on Organosolv Lignin as a Sole Carbon Source.
The complete genome sequence of the gammaproteobacterial isolate Serratia quinivorans 124R consists of 5 Mb over 2 scaffolds and a G+C content of 52.85%. Genes relating to aromatic metabolism reflect its isolation on organosolv lignin as a sole carbon source under anoxic conditions as well as the potential for lignin biorefinery applications.


ForceGen: End-to-end de novo protein generation based on nonlinear mechanical unfolding responses using a protein language diffusion model.
Through evolution, nature has presented a set of remarkable protein materials, including elastins, silks, keratins and collagens with superior mechanical performances that play crucial roles in mechanobiology. However, going beyond natural designs to discover proteins that meet specified mechanical properties remains challenging. Here we report a generative model that predicts protein designs to meet complex nonlinear mechanical property-design objectives. Our model leverages deep knowledge on protein sequences from a pretrained protein language model and maps mechanical unfolding responses to create novel proteins. Via full-atom molecular simulations for direct validation, we demonstrate that the designed proteins are novel, and fulfill the targeted mechanical properties, including unfolding energy and mechanical strength, as well as the detailed unfolding force-separation curves. Our model offers rapid pathways to explore the enormous mechanobiological protein sequence space unconstrained by biological synthesis, using mechanical features as target to enable the discovery of protein materials with superior mechanical properties.


Is a thymine dimer replicated via a transient abasic site intermediate? A comparative study using non-natural nucleotides.
UV light causes the formation of thymine dimers that can be misreplicated to induce mutagenesis and carcinogenesis. This report describes the use of a series of non-natural indolyl nucleotides in probing the ability of the high-fidelity bacteriophage T4 DNA polymerase to replicate this class of DNA lesion. Kinetic data reveal that indolyl analogues containing large pi-electron surface areas are incorporated opposite the thymine dimer almost as effectively as an abasic site, a noninstructional lesion. However, there are notable differences in the kinetic parameters for each DNA lesion that indicate distinct mechanisms for their replication. For example, the rate constants for incorporation opposite a thymine dimer are considerably slower than those measured opposite an abasic site. In addition, the magnitude of these rate constants depends equally upon contributions from pi-electron density and the overall size of the analogue. In contrast, binding of a nucleotide opposite a thymine dimer is directly correlated with the overall pi-electron surface area of the incoming dXTP. In addition to defining the kinetics of polymerization, we also provide the first reported characterization of the enzymatic removal of natural and non-natural nucleotides paired opposite a thymine dimer through exonuclease degradation or pyrophosphorolysis activity. Surprisingly, the exonuclease activity of the bacteriophage enzyme is activated by a thymine dimer but not by an abasic site. This dichotomy suggests that the polymerase can "sense" bulky lesions to partition the damaged DNA into the exonuclease domain. The data for both nucleotide incorporation and excision are used to propose models accounting for polymerase "switching" during translesion DNA synthesis.


The BCL-2 pathway preserves mammalian genome integrity by eliminating recombination-defective oocytes.
DNA double-strand breaks (DSBs) are toxic to mammalian cells. However, during meiosis, more than 200 DSBs are generated deliberately, to ensure reciprocal recombination and orderly segregation of homologous chromosomes. If left unrepaired, meiotic DSBs can cause aneuploidy in gametes and compromise viability in offspring. Oocytes in which DSBs persist are therefore eliminated by the DNA-damage checkpoint. Here we show that the DNA-damage checkpoint eliminates oocytes via the pro-apoptotic BCL-2 pathway members Puma, Noxa and Bax. Deletion of these factors prevents oocyte elimination in recombination-repair mutants, even when the abundance of unresolved DSBs is high. Remarkably, surviving oocytes can extrude a polar body and be fertilised, despite chaotic chromosome segregation at the first meiotic division. Our findings raise the possibility that allelic variants of the BCL-2 pathway could influence the risk of embryonic aneuploidy.


A qualitative assessment of 1st-year internal medicine residents' perceptions of evidence-based clinical decision making.
Residents' perceptions about evidence-based clinical decision making remain largely unexplored. To understand how residents perceive and use evidence-based medicine in clinical decision making. Qualitative study using a semistructured questionnaire and focus group in a postgraduate training program in internal medicine at an academic U.S. medical center. Seventeen 1st-year internal medicine residents in their 1st postgraduate year were interviewed. Six additional 1st-year residents formed a validation group. The interplay of time and expertise modified how physicians-in-training incorporate evidence into clinical decision making. When time was available, the residents preferred to answer their questions by searching and critically appraising the literature. This "self-acquired" expertise empowered them to help patients by using participatory decision-making styles. When time was limited, the residents turned to experts. Residents assumed that experts practiced evidence-based medicine. This "borrowed" expertise was thought to be the most efficient way of integrating evidence and clinical expertise, but it led to the use of a parental style when answers were taken back to the bedside. The practice of evidence-based medicine empowers 1st-year residents and appears to affect their choice of decision-making style. Further research is needed to better understand the link between decision-making style and evidence-based medicine.


The renin-angiotensin system and diabetic nephropathy.
The renin-angiotensin system (RAS) has key regulatory functions for blood pressure and fluid homeostasis. In addition, dysregulation of the system can have maladaptive effects to promote tissue injury in chronic diseases such as hypertension, heart failure, and kidney disease. These actions for the RAS to promote disease pathogenesis are especially apparent in diabetic nephropathy, the most common cause of end-stage renal disease in the United States. Evidence of a role for the RAS in diabetic nephropathy comes from studies in animal models and randomized clinical trials showing efficacy of angiotensin-converting enzyme inhibitors and angiotensin-receptor blockers to slow the progression of renal disease. Widespread applications of these therapies to a range of renal diseases may have contributed to the recent reduction in the incidence rates for end-stage renal disease. We provide a general review of the RAS and its role in diabetic nephropathy.


Is beauty in the face of the beholder?
Opposing forces influence assortative mating so that one seeks a similar mate while at the same time avoiding inbreeding with close relatives. Thus, mate choice may be a balancing of phenotypic similarity and dissimilarity between partners. In the present study, we assessed the role of resemblance to Self's facial traits in judgments of physical attractiveness. Participants chose the most attractive face image of their romantic partner among several variants, where the faces were morphed so as to include only 22% of another face. Participants distinctly preferred a "Self-based morph" (i.e., their partner's face with a small amount of Self's face blended into it) to other morphed images. The Self-based morph was also preferred to the morph of their partner's face blended with the partner's same-sex "prototype", although the latter face was ("objectively") judged more attractive by other individuals. When ranking morphs differing in level of amalgamation (i.e., 11% vs. 22% vs. 33%) of another face, the 22% was chosen consistently as the preferred morph and, in particular, when Self was blended in the partner's face. A forced-choice signal-detection paradigm showed that the effect of self-resemblance operated at an unconscious level, since the same participants were unable to detect the presence of their own faces in the above morphs. We concluded that individuals, if given the opportunity, seek to promote "positive assortment" for Self's phenotype, especially when the level of similarity approaches an optimal point that is similar to Self without causing a conscious acknowledgment of the similarity.


Concomitant arrhythmia surgery in patients with congenital heart disease.
Atrial tachyarrhythmia, including atrial fibrillation (AF), atrial flutter (AFL) and intra-atrial reentrant tachycardia (IART), occur frequently in patients with congenital heart disease (CHD), who may undergo multiple surgical procedures throughout life. However, data on the effectiveness of concomitant arrhythmia surgery in CHD patients are scarce. Outcome of concomitant arrhythmia surgery for AF or AFL/IART was examined in 66 successive patients [31 men (47%); age at surgery: 56 ± 14 (24-78) years] with various CHD. Concomitant arrhythmia surgery was performed in patients with a history of only AF (n = 46, 70%), only AFL/IART (n = 6, 9%) or a combination of AF and AFL/IART (n = 14, 21%). Median follow-up after arrhythmia surgery was 2 (1-4) years. AF reoccurred in 40 patients (67%), of whom 13 (22%) only had early recurrences; none of the patients with only AFL or IART prior to arrhythmia surgery developed AF after arrhythmia surgery. Recurrence-free survival of late AF was 4.6 years and differed according to the type of AF prior to surgery. Late recurrence-free survival at 3-year follow-up was 71% for paroxysmal AF, 45% for persistent AF and 20% for long-standing persistent AF (P = 0.047). Age at arrhythmia surgery was an independent predictor for late AF recurrence (odds ratio 1.05, P = 0.006). AFL/IART occurred in 17 patients (26%) after arrhythmia surgery, which was de novo in 11 patients (17%). Arrhythmia surgery in CHD patients results in freedom from late AF recurrence for a small majority of patients after median follow-up of 2 years. (Long-standing) persistent AF and older age at arrhythmia surgery are related to higher recurrence rates.


[Prevalence of overweight and obesity in children and adolescents from 1977 to 2009 - examination of the school medical data of more than 94,000 school-age children in the city of Basel (Switzerland)].
A cross-sectional study over a period of 30 years was used to investigate the development of overweight and obesity in school children. Of particular interest is the question of whether the overweight epidemic is continuing to increase, as often postulated, or whether a stagnation point or the start of a reversal of the trend has already been reached. School medical examinations take place at three school levels in the city of Basel. 94,004 student files from these examinations from the school years 1977/78 and 1978/79, as well as from 1989/90 to 2008/09, have been subjected to a secondary analysis. The overweight and obesity prevalences were calculated for eleven 2-year cohorts according to the standards of the International Obesity Taskforce IOTF. A more in-depth analysis followed, differentiated by school grade, gender and nationality. The percentage of overweight and obese children and young people has increased strongly in the last 3 decades. Whereas every 11th child was overweight 30 years ago (9.3%), it now amounts to every 5th child (20.9%). The proportion of obese children has increased from 1.2% to 5.4%. The large increase in overweight and obesity prevalences mainly took place in the 1990s. Since the millennium change, the number of overweight and obese school children among those examined has only increased slightly, or has at least no longer increased. This general development was observed in all school grades and was independent of gender and nationality, although foreign children are always twice as likely to be affected by overweight and obesity. The stabilisation can be seen at its earliest and the change in trend is clearest in the kindergarten, which could indicate a "year's cohort effect". The development over the last 10 years is welcome from a public health point of view. The continued high prevalence still represents a significant health problem, however, and the prevention efforts being made against the overweight epidemic must be further strengthened.


Both CD4(+)CD25(+) and CD4(+)CD25(-) regulatory cells mediate dominant transplantation tolerance.
CD4(+)CD25(+) T cells have been proposed as the principal regulators of both self-tolerance and transplantation tolerance. Although CD4(+)CD25(+) T cells do have a suppressive role in transplantation tolerance, so do CD4(+)CD25(-) T cells, although 10-fold less potent. Abs to CTLA-4, CD25, IL-10, and IL-4 were unable to abrogate suppression mediated by tolerant spleen cells so excluding any of these molecules as critical agents of suppression. CD4(+)CD25(+) T cells from naive mice can also prevent rejection despite the lack of any previous experience of donor alloantigens. However, this requires many more naive than tolerized cells to provide the same degree of suppression. This suggests that a capacity to regulate transplant rejection pre-exists in naive mice, and may be amplified in "tolerized" mice. Serial analysis of gene expression confirmed that cells sorted into CD4(+)CD25(+) and CD4(+)CD25(-) populations were distinct in that they responded to TCR ligation with very different programs of gene expression. Further characterization of the differentially expressed genes may lead to the development of diagnostic tests to monitor the tolerant state.


Mammaglobin peptide as a novel biomarker for breast cancer detection.
Among the different types of tests used for cancer diagnosis, molecular tests have been increrasingly incorporated because of their ability to detect either expression or functional changes in the molecules associated with the disease. Mammaglobin is a protein found in mammary tissue and can be detected in serum. This protein has been proposed as a biomarker to diagnose breast cancer, given that patients exhibit an increased amount of the protein in serum and tumor tissue, in comparison to healthy individuals. The ELISA test was used in the present study to detect mammaglobin in blood samples from 51 breast cancer patients and 51 control individuals. Antibodies against mamaglobin were generated in rabbits by using the following synthetic peptides: A (amino acids 13 to 21), B (amino acids 31 to 39), C (amino acids 56 to 64) and a D peptide, corresponding to the protein isoform without three amino acids (59, 60 and 61 amino acids) from peptide C. All peptides were immunogenic and allowed generation of antibodies that were able to discriminate patients from controls. The best results were obtained for antiserum B, achieving the best sensitivity (86.3%) and specificity (96%).


Sarcomere length-resting tension relation in single frog atrial cardiac cells.
It generally has been thought that the relatively high resting tension characteristic of cardiac tissue resides in structures (collagen, elastin) external to the individual cardiac cells, but the evidence to support this conclusion has been indirect, since the resting tension of intact single cardiac cells has not been determined previously. The purpose of the present investigation was to determine the resting tension (stress)-sarcomere length relationships of single intact frog atrial cells. For tension determinations, a single cell was attached between two poly-L-lysine coated glass beams; one beam served as a compliant calibrated cantilevered force beam, and length changes were imposed on the cell by movement of the other beam. Coventional bright-field light microscope techniques were used to view the cell, the sarcomere pattern within the cell, and the position of the force beam. The resting tension of the intact cell increased from a value of about 10 nN at a sarcomere length of 2.35 microns to a value of about 130 nN at a sarcomere length of 3.45 microns. Lagrangian and Eulerian resting stress-sarcomere length relationships were computed from the resting tension-sarcomere length relationships. The Lagrangian stress increased from a value of about 0.6 mN/mm2 at a sarcomere length of 2.35 microns to a value of about 7 mN/mm2 at a sarcomere length of 3.45 microns. These values of stress are about 8- to 30-fold less than those previously reported for intact frog atrial tissue and indicate that the resting tension of intact frog atrial preparations resides primarily in structures external to the individual cardiac cell.


A physiologically based pharmacokinetic (PBPK) approach to evaluate pharmacokinetics in patients with cancer.
Potential differences in pharmacokinetics (PK) between healthy subjects and patients with cancer were investigated using a physiologically based pharmacokinetic approach integrating demographic and physiological data from patients with cancer. Demographic data such as age, sex and body weight, and clinical laboratory measurements such as albumin, alpha-1 acid glycoprotein (AAG) and hematocrit were collected in ~2500 patients with cancer. A custom oncology population profile was built using the observed relationships among demographic variables and laboratory measurements in Simcyp® software, a population based ADME simulator. Patients with cancer were older compared with the age distribution in a built-in healthy volunteer profile in Simcyp. Hematocrit and albumin levels were lower and AAG levels were higher in patients with cancer. The custom population profile was used to investigate the disease effect on the pharmacokinetics of two probe substrates, saquinavir and midazolam. Higher saquinavir exposure was predicted in patients relative to healthy subjects, which was explained by the altered drug binding due to elevated AAG levels in patients with cancer. Consistent with historical clinical data, similar midazolam exposure was predicted in patients and healthy subjects, supporting the hypothesis that the CYP3A activity is not altered in patients with cancer. These results suggest that the custom oncology population profile is a promising tool for the prediction of PK in patients with cancer. Further evaluation and extension of this population profile with more compounds and more data will be needed.


Two-dimensional Co-Seismic Surface Displacements Field of the Chi-Chi Earthquake Inferred from SAR Image Matching.
The Mw=7.6 Chi-Chi earthquake in Taiwan occurred in 1999 over the Chelungpu fault and caused a great surface rupture and severe damage. Differential Synthetic Aperture Radar Interferometry (DInSAR) has been applied previously to study the co-seismic ground displacements. There have however been significant limitations in the studies. First, only one-dimensional displacements along the Line-of-Sight (LOS) direction have been measured. The large horizontal displacements along the Chelungpu fault are largely missing from the measurements as the fault is nearly perpendicular to the LOS direction. Second, due to severe signal decorrelation on the hangling wall of the fault, the displacements in that area are un-measurable by differential InSAR method. We estimate the co-seismic displacements in both the azimuth and range directions with the method of SAR amplitude image matching. GPS observations at the 10 GPS stations are used to correct for the orbital ramp in the amplitude matching and to create the two-dimensional (2D) co-seismic surface displacements field using the descending ERS-2 SAR image pair. The results show that the co-seismic displacements range from about -2.0 m to 0.7 m in the azimuth direction (with the positive direction pointing to the flight direction), with the footwall side of the fault moving mainly southwards and the hanging wall side northwards. The displacements in the LOS direction range from about -0.5 m to 1.0 m, with the largest displacement occuring in the northeastern part of the hanging wall (the positive direction points to the satellite from ground). Comparing the results from amplitude matching with those from DInSAR, we can see that while only a very small fraction of the LOS displacement has been recovered by the DInSAR mehtod, the azimuth displacements cannot be well detected with the DInSAR measurements as they are almost perpendicular to the LOS. Therefore, the amplitude matching method is obviously more advantageous than the DInSAR in studying the Chi-Chi earthquake. Another advantage of the method is that the displacement in the hanging wall of the fault that is un-measurable with DInSAR due to severe signal decorrelation can almost completely retrieved in this research. This makes the whole co-seismic displacements field clearly visible and the location of the rupture identifiable. Using displacements measured at 15 independent GPS stations for validation, we found that the RMS values of the differences between the two types of results were 6.9 cm and 5.7 cm respectively in the azimuth and the range directions.


Investigation into use of drugs preceding death from asthma.
Copies of death certificates were provided by the Registrar General for all deaths attributed to asthma in persons aged 5 to 34 years which were registered in England and Wales in the last quarter of 1966 and the first quarter of 1967. Information was obtained from the relevant general practitioners about 177 of the 184 subjects, and necropsy data were obtained for 113 of the 124 cases in which a post-mortem examination was known to have been made. Ninety-eight per cent. of the subjects for whom evidence was obtained were known to have been suffering from asthma, and signs of severe asthma (overdistended lungs and small bronchi plugged with mucus) were found in 91% of necropsies (57% of all deaths). Evidence that death might have been due to any other pathological condition was rare. Death was sudden and unexpected in 81% of the subjects (137 out of 171), and 59% of all deaths were referred to coroners. In 39% of cases (67 out of 171) the practitioner had not regarded the patient as suffering from severe asthma in his terminal episode. Corticosteroids and sympathomimetic preparations were the only drugs to have been used by a large proportion of patients. Two-thirds of the patients had received corticosteroids before the terminal episode, but detailed information about their use provided no suggestion that excess use could have been responsible for any large proportion of the deaths. Eighty-four per cent. of the patients were known to have used pressurized aerosol bronchodilators, and several instances of their use in excess were described. Routine inquiries about their use in the hours immediately preceding death were not made, and further evidence is required before their effect can be assessed adequately.


Force-feedback assisted and virtual fixtures based K-wire drilling simulation.
One common method to fix fractures of the human hand after an accident is an osteosynthesis with Kirschner wires (K-wires) to stabilize the bone fragments. The insertion of K-wires is a delicate minimally invasive surgery, because surgeons operate almost without a sight. Since realistic training methods are time consuming, costly and insufficient, a virtual-reality (VR) based training system for the placement of K-wires was developed. As part of this, the current work deals with the real-time bone drilling simulation using a haptic force-feedback device. To simulate the drilling, we introduce a virtual fixture based force-feedback drilling approach. By decomposition of the drilling task into individual phases, each phase can be handled individually to perfectly control the drilling procedure. We report about the related finite state machine (FSM), describe the haptic feedback of each state and explain, how to avoid jerking of the haptic force-feedback during state transition. The usage of the virtual fixture approach results in a good haptic performance and a stable drilling behavior. This was confirmed by 26 expert surgeons, who evaluated the virtual drilling on the simulator and rated it as very realistic. To make the system even more convincing, we determined real drilling feed rates through experimental pig bone drilling and transferred them to our system. Due to a constant simulation thread we can guarantee a precise drilling motion. Virtual fixtures based force-feedback calculation is able to simulate force-feedback assisted bone drilling with high quality and, thus, will have a great potential in developing medical applications.


Distribution and characterization of mosquitocidal toxin genes in some strains of Bacillus sphaericus.
The binary toxin of Bacillus sphaericus strains forms a crystal in sporulating cells, while the mosquitocidal toxin is located in the cytoplasm of vegetative cells. The distribution of binary toxin (btx) and mosquitocidal toxin (mtx) genes in 53 strains of B. sphaericus was determined by hybridization of specific gene probes to chromosomal DNA in Southern blots. btx genes were found in all strains of serotype 5a5b examined and in some strains of serotypes 1a, 3, 6, 25, and 48, while mtx genes were detected in strains of serotypes 1a, 2a2b, 5a5b, 6, 9a9c, 25, and 48. Serotype 26a26b strains lacked both toxin genes, as did some strains of serotypes 2a2b, 3, 6, and 48. Partial DNA sequences of btx genes from five strains, together with published sequences, revealed four types of toxin among mosquitocidal B. sphaericus strains. most of the 42-kDa toxin gene of btx was identical in strains from serotypes 1a, 3, 6, and 48, and the gene is here classified as a type 1 btx gene. A serotype 3 strain isolated in Singapore possessed a unique 42-kDa toxin gene, here designated type 4; while the btx genes from strains of serotypes 5a5b and 25 are referred to as types 2 and 3, respectively.


Psychosocial factors associated with depression severity in pregnant adolescents.
Adolescent depression during pregnancy is associated with increased morbidity for the teen and her infant. This cross-sectional study explored the relationships among the independent histories of alcohol use, drug use, depression, and abuse (physical or sexual) on depression severity in a diverse group of 116 pregnant adolescents (mean age = 16) who attended an urban prenatal clinic. Ever having had an alcoholic drink was a significant predictor of higher depressive scores on Children's Depression Rating Scale-Revised, β = 3.3 (0.8, 5.7); p < 0.05. History of abuse was associated with a significant 4.3-point higher mean depressive score, β = 4.3 (1.8, 6.7), p < 0.001, and remained a statistically significant predictor of more severe depressive symptoms after adjustment for history of alcohol use, history of drug use, and history of depression. This study identified that a history of physical or sexual abuse is a significant factor related to the severity of depressive symptoms in pregnant adolescents, independent of a history of alcohol, drug use, or depression. These findings suggest that an assessment of history of alcohol use, as well as abuse history, may increase the likelihood of identifying adolescents at risk for antenatal depression.


Comet Assay Profiling of FLASH-Induced Damage: Mechanistic Insights into the Effects of FLASH Irradiation.
Numerous studies have demonstrated the normal tissue-sparing effects of ultra-high dose rate 'FLASH' irradiation in vivo, with an associated reduction in damage burden being reported in vitro. Towards this, two key radiochemical mechanisms have been proposed: radical-radical recombination (RRR) and transient oxygen depletion (TOD), with both being proposed to lead to reduced levels of induced damage. Previously, we reported that FLASH induces lower levels of DNA strand break damage in whole-blood peripheral blood lymphocytes (WB-PBL) ex vivo, but our study failed to distinguish the mechanism(s) involved. A potential outcome of RRR is the formation of crosslink damage (particularly, if any organic radicals recombine), whilst a possible outcome of TOD is a more anoxic profile of induced damage resulting from FLASH. Therefore, the aim of the current study was to profile FLASH-induced damage via the Comet assay, assessing any DNA crosslink formation as a putative marker of RRR and/or anoxic DNA damage formation as an indicative marker of TOD, to determine the extent to which either mechanism contributes to the "FLASH effect". Following FLASH irradiation, we see no evidence of any crosslink formation; however, FLASH irradiation induces a more anoxic profile of induced damage, supporting the TOD mechanism. Furthermore, treatment of WB-PBLs pre-irradiation with BSO abrogates the reduced strand break damage burden mediated by FLASH exposures. In summary, we do not see any experimental evidence to support the RRR mechanism contributing to the reduced damage burden induced by FLASH. However, the observation of a greater anoxic profile of damage following FLASH irradiation, together with the BSO abrogation of the reduced strand break damage burden mediated by FLASH, lends further support to TOD being a driver of the reduced damage burden plus a change in the damage profile mediated by FLASH.


Multiobjective Evolutionary Algorithms Are Still Good: Maximizing Monotone Approximately Submodular Minus Modular Functions.
As evolutionary algorithms (EAs) are general-purpose optimization algorithms, recent theoretical studies have tried to analyze their performance for solving general problem classes, with the goal of providing a general theoretical explanation of the behavior of EAs. Particularly, a simple multiobjective EA, that is, GSEMO, has been shown to be able to achieve good polynomial-time approximation guarantees for submodular optimization, where the objective function is only required to satisfy some properties and its explicit formulation is not needed. Submodular optimization has wide applications in diverse areas, and previous studies have considered the cases where the objective functions are monotone submodular, monotone non-submodular, or non-monotone submodular. To complement this line of research, this article studies the problem class of maximizing monotone approximately submodular minus modular functions (i.e., g-c) with a size constraint, where g is a so-called non-negative monotone approximately submodular function and c is a so-called non-negative modular function, resulting in the objective function (g-c) being non-monotone non-submodular in general. Different from previous analyses, we prove that by optimizing the original objective function (g-c) and the size simultaneously, the GSEMO fails to achieve a good polynomial-time approximation guarantee. However, we also prove that by optimizing a distorted objective function and the size simultaneously, the GSEMO can still achieve the best-known polynomial-time approximation guarantee. Empirical studies on the applications of Bayesian experimental design and directed vertex cover show the excellent performance of the GSEMO.


Risk Factors for Intraoperative Periprosthetic Femoral Fractures During Primary Total Hip Arthroplasty. An Analysis From the National Joint Registry for England and Wales and the Isle of Man.
The aim of this study is to estimate risk factors for intraoperative periprosthetic femoral fractures (IOPFF) and each anatomic subtype (calcar crack, trochanteric fracture, femoral shaft fracture) during primary total hip arthroplasty. This retrospective cohort study included 793,823 primary total hip arthroplasties between 2004 and 2016. Multivariable regression modeling was used to estimate relative risk of patient, surgical, and implant factors for any IOPFF and for all anatomic subtypes of IOPFF. Clinically important interactions were assessed using multivariable regression. Patient factors significantly increasing the risk of fracture were female gender, American Society of Anesthesiologists grade 3 to 5, and preoperative diagnosis including avascular necrosis of the hip, previous trauma, inflammatory disease, pediatric disease, and previous infection. Overall risk of IOPFF associated with age was greatest in patients below 50 years and above 80 years. Risk of any fracture reduced with computer-guided surgery and in non-National Health Service hospitals. Nonposterior approaches increased the risk of shaft and trochanteric fracture only. Cementless implants significantly increased the risk of only calcar cracks and shaft fractures and not trochanteric fractures. Fracture risk increases in patients younger than 50 and older than 80 years, females, American Society of Anesthesiologists grade 3 to 5, and indications other than primary osteoarthritis. Large cumulative reduction in IOPFF risk may occur with use of cemented implants, posterior approach, and computer-guided surgery. Level 3b (cohort study).


Fat fibrosis: friend or foe?
At the simplest level, obesity is the manifestation of an imbalance between caloric intake and expenditure; however, the pathophysiological mechanisms that govern the development of obesity and associated complications are enormously complex. Fibrosis within the adipose tissue compartment is one such factor that may influence the development of obesity and/or obesity-related comorbidities. Furthermore, the functional consequences of adipose tissue fibrosis are a matter of considerable debate, with evidence that fibrosis serves both adaptive and maladaptive roles. Tissue fibrosis itself is incompletely understood, and multiple cellular and molecular pathways are involved in the development, maintenance, and resolution of the fibrotic state. Within the context of obesity, fibrosis influences molecular and cellular events that relate to adipocytes, inflammatory cells, inflammatory mediators, and supporting adipose stromal tissue. In this Review, we explore what is known about the interplay between the development of adipose tissue fibrosis and obesity, with a view toward future investigative and therapeutic avenues.


Effect of thermal processing on the flavonols rutin and quercetin.
The current research involves the study of the thermal treatment of quercetin and rutin in an aqueous model system (cooking). These substances were heated and their degradation was followed by high-performance liquid chromatography/diode-array detection (HPLC/DAD). The influence of pH and the involvement of oxygen in the degradation were studied. HPLC/electrospray ionization multi-stage mass spectrometry (ESI-MS(n)) was used for the structural characterization of the compounds produced. The influence of the degradation of the phenolic compounds on their antioxidant properties was elucidated by a electron spin resonance (ESR) spectrometry study of the reaction samples mixed with the stabilized radical, Fremy's salt. Strong degradation of the model substances took place under weak basic and oxidative conditions. Quercetin showed the most intense degradation. Protocatechuic acid could be identified as a cleavage reaction product by analyzing its retention time and molar mass during the degradation of quercetin. The structure of a second cleavage product could be identified on the basis of ESI-MS(n) fragmentation data. Also, several structures for reaction products of oxidized quercetin are suggested. The ESR analysis showed a decrease in the antioxidant activity of the reaction samples after heat treatment in aqueous solution.


Cytotoxic diterpenoids from Euphorbia helioscopia.
Four new jatrophane-type diterpenoids (1-4), together with 16 known related compounds, were isolated from the Chinese medicinal plant Euphorbia helioscopia. The structures of 1-4 were determined on the basis of spectroscopic and chemical methods. Cytotoxicity of the isolated compounds against HeLa and MDA-MB-231 cells was evaluated, with only helioscopinolide A (5) and euphornin (3a) being active.


A clinical "near miss" highlights risk management issues surrounding ultrasound-guided and wire-localised breast resections.
The introduction of the National Health Service (NHS) Breast Screening Programme has led to a considerable increase in the detection of impalpable breast cancer. Patients with impalpable breast cancer typically undergo oncological resection facilitated either by the insertion of guide wires placed stereo-tactically or through ultra-sound guided skin markings to delineate the extent of a lesion. The need for radiological interventions on the day of surgery adds complexity and introduces the risk that a patient may accidentally transferred to the operating room directly without the image guidance procedure. A case is described of a patient who required a pre-operative ultrasound scan in order to localise an impalpable breast cancer but who was accidentally taken directly to the operating theatre (OR) and anaesthetised without pre-operative intervention. The radiologist was called to the OR and an on-table ultrasound was performed without further consequence. It is evident that breast cancer patients undergoing image-guided resection are exposed to an additional layer of clinical risks. These risks are not offset by the World Health Organisation surgical safety checklist in its present guise. Here, we review a number of simple and inexpensive changes to the system that may improve the safety of the breast cancer patient undergoing surgery.


Association of energy intake and energy balance with postmenopausal breast cancer in the prostate, lung, colorectal, and ovarian cancer screening trial.
Energy restriction remains one of the most effective ways known to prevent breast cancer in animal models. However, energy intake has not been consistently associated with risk of breast cancer in humans. In a prospective study, we assessed whether energy intake, body size, and physical activity each independently influence breast cancer risk in postmenopausal women and estimated the joint effect of combinations of these individual factors. As part of the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial, 38,660 women, ages 55 to 74 years and recruited from 10 centers in the United States during 1993 to 2001, were randomized to the screening arm of the trial. At baseline, the women completed a self-administered questionnaire, including a food frequency questionnaire. During follow-up from 1993 to 2003, 764 incident breast cancer cases were ascertained. Women in the highest quartile of energy intake (> or = 2,084 kcal/d) compared with those in the lowest quartile (<1,316 kcal/d) had a significantly increased risk for breast cancer [multivariate relative risk (RR), 1.25; 95% confidence interval (95% CI), 1.02-1.53; P(trend continuous) = 0.03]. Current body mass index (BMI) was also positively and significantly associated with risk (multivariate RR comparing >30 kg/m2 with <22.5 kg/m2, 1.35; 95% CI, 1.06-1.70; P(trend) = 0.01). Women with > or = 4 hours/wk of vigorous recreational physical activity had a significantly reduced risk of breast cancer compared with those who reported no recreational physical activity (multivariate RR, 0.78; 95% CI, 0.60-0.99; P(trend) = 0.15). None of these associations with individual energy balance measures was substantially confounded by the other two measures. When we estimated the joint effect of all three variables, women with the most unfavorable energy balance (the highest energy intake, highest BMI, and least physical activity) had twice the risk (RR, 2.10; 95% CI, 1.27-3.45) of women with the most favorable energy balance (the lowest energy intake, lowest BMI, and most physical activity). Although our estimates of absolute energy intake, based on a food frequency questionnaire, are imperfect, these results suggest that energy intake, in addition to BMI and physical activity may be independently associated with breast cancer risk. In addition, these three aspects of energy balance may act jointly in determining breast cancer risk.


Cancer mortality and morbidity in employees of the United Kingdom Atomic Energy Authority, 1946-86.
In further analyses of a cohort of 39,718 United Kingdom Atomic Energy Authority employees after 7 more years follow-up, cancer mortality, based on 1,506 deaths in 1946-86, was 20% below the national average. Prostatic cancer mortality showed a statistically significant association with external radiation exposure, largely confined to men who were also monitored for internal contamination by radionuclides other than plutonium. Prostatic cancer mortality was highest in radiation workers at Winfrith. In women monitored for radiation exposure, mortality from cancer of the uterus (including the cervix uteri) was increased relative to other employees, and, showed a statistically significant association with external radiation exposure. While there were some other statistically significant results, as would be expected by chance alone when multiple comparisons are made, there were no other cancer sites with consistently exceptional findings. Point estimates for risk associated with increasing exposure to radiation suggest a decrease of four deaths per 10(4) person-years per Sv for leukaemia and an increase of 20 deaths for all cancers except leukaemia, but confidence intervals indicate that a wide range of values are compatible with the data. Cancer morbidity based on 1,699 registrations in 1971-84 was 12% below the national average. Findings from site-specific analyses largely replicated those of the mortality analyses.


Natural killer cells activated in a human mixed lymphocyte response culture identified by expression of Leu-11 and class II histocompatibility antigens.
Lymphoid cells with natural killer (NK)-like function, morphology, and antigenic phenotype have been identified in a mixed lymphocyte culture generated by co-culture of human peripheral blood mononuclear cells with an allogeneic human B lymphoblastoid cell line CCRF-SB. The majority of these mixed lymphocyte (MLR)-response activated NK cells express the Leu-11 surface antigen, but do not express certain T cell-associated antigens (Leu-1, Leu-3, and Leu-4) or the mature monocyte specific antigen, Leu-M3. Unlike most freshly isolated Leu-11+ human NK cells, the MLR-activated Leu-11+ cells expressed class II major histocompatibility antigens, DR and DC. Concomitant with expression of class II gene products, the Leu-11+,DR+ NK cells demonstrate enhanced cytotoxicity against the NK-sensitive tumor cell line K562. The presence of mitotic cells in the Leu-11+,DR+ population and the acquisition of increased levels of transferrin receptor on the cell surface were further indicators of activation of these cells. The direct precursors of the MLR-activated Leu-11+,DR+ cell were Leu-11+ cells that lacked expression of another NK-associated antigen Leu-7, i.e., Leu-7-,11+. These studies provided a definitive identification of the "NK-like" cell in MLR cultures and thus allow quantitation and isolation of these cells for further study.


Validation of an early warning score in pre-intensive care unit.
Prognosis of patients in the intensive care unit (ICU) has a relation with their severity just before admission. The Modified Early Warning Score (MEWS) was used to evaluate the severe condition of patients 12, 24 and 72 hours before admission in the ICU, assess the most prevalent parameters and correlate the MEWS before ICU with the outcome (survival versus death). Retrospective analyses of 65 patients consecutively admitted to the ICU from July to October, 2006 evaluating the physiological parameters 72 hours prior to admission. APACHE II mean was 22.2 ± 7.9 points, mortality was 54.6% and standardized mortality ratio means was 1.24. MEWS means were 3.7 ± 0.2; 4.0 ± 0.2 and 5.1 ± 0.2 points, calculated 72, 48 and 24 hours previous to ICU admission, respectively. An increasing percentage of patients with MEWS > 3 points within 72, 48 and 24 hours before admission - 43.8%, 59.4% and 73.4%, respectively was recorded. Among the included physiological parameters respiratory rate contributed the most to the MEWS. Highest mortality was found in patients with MEWS > 3 points already found 72 hours before admission. Patients who died presented with a significant increase in the MEWS 24 hours prior to admission to the ICU (in relation to the MEWS recorded 72 hours before) but the situation was not identified in survivors. MEWS closely identified the severity of patients admitted to the ICU, suggesting that it can be a reliable score, useful in the situations preceding the ICU.


MULTIDETECTOR-ROW COMPUTED TOMOGRAPHY PATTERNS OF BRONCHOESPHAGEAL ARTERY HYPERTROPHY AND SYSTEMIC-TO-PULMONARY FISTULA IN DOGS.
Anomalies involving arterial branches in the lungs are one of the causes of hemoptysis in humans and dogs. Congenital and acquired patterns of bronchoesophageal artery hypertrophy have been reported in humans based on CT characteristics. The purpose of this retrospective study was to describe clinical, echocardiographic, and multidetector computed tomography features of bronchoesophageal artery hypertrophy and systemic-to-pulmonary arterial communications in a sample of 14 dogs. Two main vascular patterns were identified in dogs that resembled congenital and acquired conditions reported in humans. Pattern 1 appeared as an aberrant origin of the right bronchoesophageal artery, normal origin of the left one, and enlargement of both the bronchial and esophageal branches that formed a dense network terminating in a pulmonary artery through an orifice. Pattern 2 appeared as a normal origin of both right and left bronchoesophageal arteries, with an enlarged and tortuous course along the bronchi to the periphery of the lung, where they communicated with subsegmental pulmonary arteries. Dogs having Pattern 1 also had paraesophageal and esophageal varices, with the latter being confirmed by videoendoscopy examination. Authors conclude that dogs with Pattern 1 should be differentiated from dogs with other congenital vascular systemic-to-pulmonary connections. Dogs having Pattern 2 should be evaluated for underlying pleural or pulmonary diseases. Bronchoesophageal artery hypertrophy can be accompanied by esophageal venous engorgement and should be included in the differential diagnosis for esophageal and paraesophageal varices in dogs.


Herbal supplement extends life span under some environmental conditions and boosts stress resistance.
Genetic studies indicate that aging is modulated by a great number of genetic pathways. We have used Drosophila longevity and stress assays to test a multipath intervention strategy. To carry out this strategy, we supplemented the flies with herbal extracts (SC100) that are predicted to modulate the expression of many genes involved in aging and stress resistance, such as mTOR, NOS, NF-KappaB, and VEGF. When flies were housed in large cages with SC100 added, daily mortality rates of both male and female flies were greatly diminished in mid to late life. Surprisingly, SC100 also stabilized midlife mortality rate increases so as to extend the maximum life span substantially beyond the limits previously reported for D. melanogaster. Under these conditions, SC100 also promoted robust resistance to partial starvation stress and to heat stress. Fertility was the same initially in both treated and control flies, but it became significantly higher in treated flies at older ages as the fertility of control flies declined. Mean and maximum life spans of flies in vials at the same test site were also extended by SC100, but the life spans were short in absolute terms. In contrast, at an independent test site where stress was minimized, the flies exhibited much longer mean life spans, but the survival curves became highly rectangular and the effects of SC100 on both mean and maximum life spans declined greatly or were abolished. The data indicate that SC100 is a novel herbal mix with striking effects on enhancing Drosophila stress resistance and life span in some environments, while minimizing mid to late life mortality rates. They also show that the environment and other factors can have transformative effects on both the length and distribution of survivorship, and on the ability of SC100 to extend the life span.


The Botrytis cinerea Gene Expression Browser.
For comprehensive gene expression analyses of the phytopathogenic fungus Botrytis cinerea, which infects a number of plant taxa and is a cause of substantial agricultural losses worldwide, we developed BEB, a web-based B. cinerea gene Expression Browser. This computationally inexpensive web-based application and its associated database contain manually curated RNA-Seq data for B. cinerea. BEB enables expression analyses of genes of interest under different culture conditions by providing publication-ready heatmaps depicting transcript levels, without requiring advanced computational skills. BEB also provides details of each experiment and user-defined gene expression clustering and visualization options. If needed, tables of gene expression values can be downloaded for further exploration, including, for instance, the determination of differentially expressed genes. The BEB implementation is based on open-source computational technologies that can be deployed for other organisms. In this case, the new implementation will be limited only by the number of transcriptomic experiments that are incorporated into the platform. To demonstrate the usability and value of BEB, we analyzed gene expression patterns across different conditions, with a focus on secondary metabolite gene clusters, chromosome-wide gene expression, previously described virulence factors, and reference genes, providing the first comprehensive expression overview of these groups of genes in this relevant fungal phytopathogen. We expect this tool to be broadly useful in B. cinerea research, providing a basis for comparative transcriptomics and candidate gene identification for functional assays.


Cannabidiol Induces Cell Cycle Arrest and Cell Apoptosis in Human Gastric Cancer SGC-7901 Cells.
The main chemical component of cannabis, cannabidiol (CBD), has been shown to have antitumor properties. The present study examined the in vitro effects of CBD on human gastric cancer SGC-7901 cells. We found that CBD significantly inhibited the proliferation and colony formation of SGC-7901 cells. Further investigation showed that CBD significantly upregulated ataxia telangiectasia-mutated gene (ATM) and p53 protein expression and downregulated p21 protein expression in SGC-7901 cells, which subsequently inhibited the levels of CDK2 and cyclin E, thereby resulting in cell cycle arrest at the G0-G1 phase. In addition, CBD significantly increased Bax expression levels, decreased Bcl-2 expression levels and mitochondrial membrane potential, and then upregulated the levels of cleaved caspase-3 and cleaved caspase-9, thereby inducing apoptosis in SGC-7901 cells. Finally, we found that intracellular reactive oxygen species (ROS) increased after CBD treatment. These results indicated that CBD could induce G0-G1 phase cell cycle arrest and apoptosis by increasing ROS production, leading to the inhibition of SGC-7901 cell proliferation, thereby suggesting that CBD may have therapeutic effects on gastric cancer.


Understanding and interpreting serum protein electrophoresis.
Serum protein electrophoresis is used to identify patients with multiple myeloma and other serum protein disorders. Electrophoresis separates proteins based on their physical properties, and the subsets of these proteins are used in interpreting the results. Plasma protein levels display reasonably predictable changes in response to acute inflammation, malignancy, trauma, necrosis, infarction, burns, and chemical injury. A homogeneous spike-like peak in a focal region of the gamma-globulin zone indicates a monoclonal gammopathy. Monoclonal gammopathies are associated with a clonal process that is malignant or potentially malignant, including multiple myeloma, Waldenstrom's macroglobulinemia, solitary plasmacytoma, smoldering multiple myeloma, monoclonal gammopathy of undetermined significance, plasma cell leukemia, heavy chain disease, and amyloidosis. The quantity of M protein, the results of bone marrow biopsy, and other characteristics can help differentiate multiple myeloma from the other causes of monoclonal gammopathy. In contrast, polyclonal gammopathies may be caused by any reactive or inflammatory process.


Lack of evidence for close linkage of the glutamate GluR6 receptor gene with schizophrenia.
Previous research has consistently implicated genetic factors in the pathogenesis of schizophrenia. It has been hypothesized that an abnormality in glutamatergic function is of etiologic importance in schizophrenia, and therefore the glutamate receptor family of genes are potential susceptibility loci for schizophrenia. To test this hypothesis the authors sought to detect linkage between the GluR6 glutamate receptor gene and schizophrenia. Twenty-three English and Icelandic families containing multiple cases of schizophrenia were genotyped with a microsatellite trinucleotide repeat polymorphism localized at the GluR6 glutamate receptor locus. Lod scores, model-free linkage analysis, and extended relative pair analysis were used to test for linkage. No statistically significant evidence of linkage between GluR6 and schizophrenia was found. The results do not support the hypothesis that GluR6 allelic variants provide a major gene contribution to the etiology of schizophrenia in a large proportion of these pedigrees.


First Report in Mali of Xanthomonas citri pv. mangiferaeindicae Causing Mango Bacterial Canker on Mangifera indica.
Bacterial canker (or black spot) of mango caused by Xanthomonas citri pv. mangiferaeindicae is an important disease in tropical and subtropical areas (1). X. citri pv. mangiferaeindicae can cause severe infection in a wide range of mango cultivars and induces raised, angular, black leaf lesions, sometimes with a chlorotic halo. Severe leaf infection may result in abscission. Fruit symptoms appear as small, water-soaked spots on the lenticels that later become star shaped, erumpent, and exude an infectious gum. Often, a "tear stain" infection pattern is observed on the fruit. Severe fruit infections cause premature drop. Twig cankers are potential sources of inoculum and weaken branch resistance to winds. Yield loss up to 85% has been reported at grove scale for susceptible cultivars (1). Suspected leaf lesions of bacterial canker were collected in July 2010 from mango trees in four, six, and three localities of the Koulikoro, Sikasso, and Bougouni provinces of Mali, respectively (i.e., the major mango-growing areas in this country). Nonpigmented Xanthomonas-like colonies were isolated on KC semiselective medium (3). Twenty-two strains from Mali were identified as X. citri pv. mangiferaeindicae based on IS1595-ligation-mediated PCR (4) and they produced fingerprints fully identical to that of strains isolated from Ghana and Burkina Faso. Five Malian strains (LH409, LH410, LH414, LH415-3, and LH418) were compared by multilocus sequence analysis (MLSA) to the type strain of X. citri and the pathotype strain of several X. citri pathovars, including pvs. anacardii and mangiferaeindicae. This assay targeted the atpD, dnaK, efp, and gyrB genes, as described previously (2). Nucleotide sequences were 100% identical to those of the pathotype strain of X. citri pv. mangiferaeindicae whatever the gene assayed, but differed from any other assayed X. citri pathovar. Leaves of mango cv. Maison Rouge from the youngest vegetative flush were infiltrated (10 inoculation sites per leaf for three replicate leaves on different plants per bacterial strain) with the same five strains from Mali. Bacterial suspensions (~1 × 105 CFU/ml) were prepared in 10 mM Tris buffer (pH 7.2) from 16-h-old cultures on YPGA (7 g of yeast, 7 g of peptone, 7 g of glucose, and 18 g of agar/liter, pH 7.2). The negative control treatment consisted of three leaves infiltrated with sterile Tris buffer (10 sites per leaf). Plants were incubated in a growth chamber at 30 ± 1°C by day and 26 ± 1°C by night (12-h/12-h day/night cycle) at 80 ± 5% relative humidity. All leaves inoculated with the Malian strains showed typical symptoms of bacterial canker a week after inoculation. No lesions were recorded from the negative controls. One month after inoculation, mean X. citri pv. mangiferaeindicae population sizes ranging from 5 × 106 to 1 × 107 CFU/lesion were recovered from leaf lesions, typical of a compatible interaction (1). To our knowledge, this is the first report of the disease in Mali. Investigations from local growers suggest that the disease may have been present for some years in Mali but likely less than a decade. A high disease incidence and severity were observed, suggesting the suitability of environmental conditions in this region for the development of mango bacterial canker. References: (1) N. Ah-You et al. Phytopathology 97:1568, 2007. (2) L. Bui Thi Ngoc et al. Int. J. Syst. Evol. Microbiol. 60:515, 2010. (3) O. Pruvost et al. J. Appl. Microbiol. 99:803, 2005. (4) O. Pruvost et al. Phytopathology 101:887, 2011.


Sex differences in heel pad stiffness during in vivo loading and unloading.
Due to conflicting data from previous studies a new methodological approach to evaluate heel pad stiffness and soft tissue deformation has been developed. The purpose of this study was to compare heel pad (HP) stiffness in both limbs between males and females during a dynamic unloading and loading activity. Ten males and 10 females volunteered to perform three dynamic trials to unload and load the HP. The dynamic protocol consisted of three continuous phases: foot flat (baseline phase), bilateral heel raise (unloading phase) and foot flat (loading phase) with each phase lasting two seconds. Six retroreflective markers (3 mm) were attached to the skin of the left and right heels using a customised marker set. Three-dimensional motion analysis cameras synchronised with force plates collected the kinematic and kinetic data throughout the trials. Three-way repeated measures ANOVA together with a Bonferroni post hoc test were applied to the stiffness and marker displacement datasets. On average, HP stiffness was higher in males than females during the loading and unloading phases. ANOVA results revealed no significant differences for the stiffness and displacement outputs with respect to sex, sidedness or phase interactions (p > .05) in the X, Y and Z directions. Irrespective of direction, there were significant differences in stiffness between the baseline and unloading conditions (p < .001) but no significant differences between the baseline and loaded conditions (p = 1.000). Post hoc analyses for the marker displacement showed significant differences between phases for the X and Z directions (p < .032) but no significant differences in the Y direction (p > .116). Finally, females portrayed lower levels of mean HP stiffness whereas males had stiffer heels particularly in the vertical direction (Z) when the HP was both unloaded and loaded. High HP stiffness values and very small marker displacements could be valuable indicators for the risk of pathological foot conditions.


Use of the immunoglobulin G avidity assay to differentiate between recent Zika and past dengue virus infections.
Zika (ZIKV) and dengue (DENV) virus infections elicit a robust but cross-reactive antibody response against the viral envelope protein, while antibody responses against non-structural proteins (NS) are more virus specific. Building on this premise, we have previously developed a flavivirus multiplex microsphere immunoassay (MIA) for the serologic diagnosis of ZIKV and DENV infections. This assay significantly improved diagnostic accuracy; however, MIA could not differentiate more recent from past infections, which still represents a major diagnostic challenge. Therefore, an immunoglobulin G (IgG) based avidity assay was developed and its diagnostic performance evaluated. Specimens from New York State residents were submitted to the Wadsworth Center New York State Department of Health (NYSDOH) for routine clinical testing by Zika IgM ELISA and plaque reduction neutralization test (PRNT). Using our previously developed flavivirus MIA as a platform, we developed an IgG avidity assay to discriminate recent ZIKV from past DENV infections. Zika IgM positive specimens had an average Zika IgG avidity index of 14.8% (95% CI: 11.0-18.4%), while Zika IgM negative but flavivirus MIA and PRNT positive samples had an average Zika IgG avidity index of 34.9% (95% CI: 31.1-38.7%). Specimens positive for dengue antibodies by flavivirus MIA and PRNT had an average dengue IgG avidity index of 68.7% (95% CI: 62.7-75.0%). The IgG avidity assay accurately distinguished recent ZIKV from past DENV infections in patients who traveled to dengue endemic regions. This assay could be very useful in patients with high risk of Zika complications such as pregnant women and monitoring immune responses in vaccine trials.


Stent-assisted detachable coil embolization of wide-necked renal artery aneurysms.
Renal artery aneurysms (RAAs) are rare with an estimated incidence of 0.1% in the general population, and they represent approximately 25% of all visceral aneurysms. The gold standard of treatment is open surgery, but it is associated with a high risk of nephrectomy, mortality, and morbidity. Less invasive endovascular therapies are becoming increasingly common for the treatment of RAAs. Here, we aimed to report three cases of wide-necked complex renal artery aneurysms treated endovascularly using stent-assisted coil embolization with self-expandable stent nitinol Solitaire AB and Concerto Axium coils. In addition, we describe the use of the waffle-cone technique in a case of wide-necked saccular RAA involving the renal artery bifurcation. Technical success was achieved in all three cases with no early or late complications and no recurrences.


Extensive SUMO Modification of Repressive Chromatin Factors Distinguishes Pluripotent from Somatic Cells.
Post-translational modification by SUMO is a key regulator of cell identity. In mouse embryonic fibroblasts (MEFs), SUMO impedes reprogramming to pluripotency, while in embryonic stem cells (ESCs), it represses the emergence of totipotent-like cells, suggesting that SUMO targets distinct substrates to preserve somatic and pluripotent states. Using MS-based proteomics, we show that the composition of endogenous SUMOylomes differs dramatically between MEFs and ESCs. In MEFs, SUMO2/3 targets proteins associated with canonical SUMO functions, such as splicing, and transcriptional regulators driving somatic enhancer selection. In contrast, in ESCs, SUMO2/3 primarily modifies highly interconnected repressive chromatin complexes, thereby preventing chromatin opening and transitioning to totipotent-like states. We also characterize several SUMO-modified pluripotency factors and show that SUMOylation of Dppa2 and Dppa4 impedes the conversion to 2-cell-embryo-like states. Altogether, we propose that rewiring the repertoire of SUMO target networks is a major driver of cell fate decision during embryonic development.


Investigation of the mechanism of nicotine-induced relaxation on the sheep sphincter of Oddi.
Possible mechanisms for nicotine-induced relaxation were investigated in the isolated sheep's sphincter of Oddi. Sheep's sphincter of Oddi rings were mounted in tissue bath with modified Krebs-Henseleit solution and aerated with 95% oxygen and 5% carbon dioxide. Tension was measured with isometric force transducers, and muscle relaxation was expressed as percent decrease of precontraction induced by carbachol. Nicotine (1 x 10(-5) to 3 x 10(-3) mol/L) produced concentration-dependent relaxation on sphincter of Oddi precontracted by carbachol (10(-6) mol/L). Nicotine-induced relaxation was 72.8 +/- 4.2% of precontraction with carbachol (10(-6) mol/L) (mean pD2 value, 3.76 +/- 0.05 mol/L). Nicotine-induced relaxation was not affected by N(w)-nitro L-arginine methyl ester (L-NAME) (3 x 10(-5) mol/L), methylene blue (10(-5) mol/L), indomethacin (10(-5) mol/L), hexamethonium (10(-5) mol/L), glibenclamide (10(-5) mol/L), 4-aminopyridine (10(-3) mol/L), tetraethylammonium (3 x 10(-4) mol/L), clotrimazole (10(-6) mol/L), 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) (10(-6) mol/L), and anthracene-9-carboxylate (9-AC) (10(-6) mol/L), but potentiated by bupivacain (10(-5) mol/L). A calcium-antagonizing effect of nicotine was not observed. The results suggest that nicotine-induced relaxation of the sheep's sphincter of Oddi is not mediated by the release of prostaglandins, nitric oxide (NO), or a related substance; by the activation of potassium channels or chloride channels; or by the stimulation of nicotinic cholinoceptors. Potentiation of the nicotine-induced relaxation by bupivacain indicates that blockade of sodium channels may play a role in this relaxation.


Semicircular canal morphogenesis in the zebrafish inner ear requires the function of gpr126 (lauscher), an adhesion class G protein-coupled receptor gene.
Morphogenesis of the semicircular canal ducts in the vertebrate inner ear is a dramatic example of epithelial remodelling in the embryo, and failure of normal canal development results in vestibular dysfunction. In zebrafish and Xenopus, semicircular canal ducts develop when projections of epithelium, driven by extracellular matrix production, push into the otic vesicle and fuse to form pillars. We show that in the zebrafish, extracellular matrix gene expression is high during projection outgrowth and then rapidly downregulated after fusion. Enzymatic disruption of hyaluronan in the projections leads to their collapse and a failure to form pillars: as a result, the ears swell. We have cloned a zebrafish mutant, lauscher (lau), identified by its swollen ear phenotype. The primary defect in the ear is abnormal projection outgrowth and a failure of fusion to form the semicircular canal pillars. Otic expression of extracellular matrix components is highly disrupted: several genes fail to become downregulated and remain expressed at abnormally high levels into late larval stages. The lau mutations disrupt gpr126, an adhesion class G protein-coupled receptor gene. Expression of gpr126 is similar to that of sox10, an ear and neural crest marker, and is partially dependent on sox10 activity. Fusion of canal projections and downregulation of otic versican expression in a hypomorphic lau allele can be restored by cAMP agonists. We propose that Gpr126 acts through a cAMP-mediated pathway to control the outgrowth and adhesion of canal projections in the zebrafish ear via the regulation of extracellular matrix gene expression.


Understanding Contextual Differences in Residential LTC Provision for Cross-National Research: Identifying Internationally Relevant CDEs.
Long-term care (LTC) reflects a growing emphasis on person-centered care (PCC), with services oriented around individuals' needs and preferences. Addressing contextual and cultural differences across countries offers important insight into factors that facilitate or hinder application of PCC practices within and across countries. This article takes an international lens to consider country-specific contexts of LTC, describing preliminary steps to develop common data elements that capture contextual differences across LTC settings globally. Through an iterative series of online, telephone, and in-person sessions, we engaged in in-depth discussions with 11 colleague experts in residential LTC and coauthors from six countries (China and Hong Kong, England, Sweden, Thailand, Trinidad and Tobago, and the United States). Our discussions yielded rich narrative describing a vast range in types of LTC settings, leading to our development of a working definition of residential LTC. Scope of services, funding, ownership, and regulations varied greatly across countries and across different residential LTC settings within countries. Moving forward, we recommend expanding our activities to countries that reflect different stages of residential LTC development. Our goal is to contribute to a larger initiative underway by the WE-THRIVE consortium to establish a global research measurement infrastructure that advances PCC internationally.


Determinants of tension-type headache in children.
The objective of this study was to study the prevalence, characteristics and predisposing factors of tension-type headache in children. An unselected population-based questionnaire study was carried out in 1409 Finnish schoolchildren aged 12 years. Of them, 1135 (81%) returned an acceptably completed questionnaire. The prevalence of episodic tension-type headache in children was 12% (138 of 1135). Children with episodic tension-type headache also often reported characteristics of pain typical for migraine. Children with frequent and persistent episodic tension-type headache reported stabbing and severe occipital pain, phonophobia and abdominal pain significantly more often than children with infrequent episodic tension-type headache. Neck-shoulder symptoms, symptoms of depression and oromandibular dysfunction were each independently associated with episodic tension-type headache. The father's occupation of a lower-level white-collar worker put the child at a four-fold risk for episodic tension-type headache. We conclude that episodic tension-type headache is as common as migraine in children. It can be associated with depression, oromandibular dysfunction and muscular stress. Especially children with frequent and persistent episodic tension-type headache report characteristics of pain typical for migraine.


School recess and group classroom behavior.
This study examines the amount of recess that children 8 to 9 years of age receive in the United States and compares the group classroom behavior of children receiving daily recess with that of children not receiving daily recess. This is a secondary analysis of a public-use data set, the Early Childhood Longitudinal Study, Kindergarten Class of 1998-1999, third-grade data set. Children were categorized into 2 levels of recess exposure, that is, none/minimal break (<1 break of 15 minutes/day) or some recess. Some recess was further categorized into 5 levels on the basis of frequency and duration of recess. Child, parent, school, and classroom characteristics of those with and without recess were compared. The group classroom behavior was assessed by using the teacher's rating of class behavior. Complete data were available for 10301 to 11624 children 8 to 9 years of age. There were equal numbers of boys and girls (boys: 50.3%). Children exposed to none/minimal break (30%) were much more likely to be black, to be from families with lower incomes and lower levels of education, to live in large cities, to be from the Northeast or South, and to attend public school, compared with those with recess. Teacher's rating of classroom behavior scores were better for children with some recess than for those with none/minimal break. This finding was maintained in multivariate regression analysis. However, among children receiving daily recess, the teacher's rating of class behavior scores did not differ significantly according to the level of exposure. These results indicated that, among 8- to 9-year-old children, having > or =1 daily recess period of >15 minutes in length was associated with better teacher's rating of class behavior scores. This study suggests that schoolchildren in this age group should be provided with daily recess.


Osteogenesis imperfecta: from diagnosis and multidisciplinary treatment to future perspectives.
Osteogenesis imperfecta is an inherited connective tissue disorder with wide phenotypic and molecular heterogeneity. A common issue associated with the molecular abnormality is a disturbance in bone matrix synthesis and homeostasis inducing bone fragility. In very early life, this can lead to multiple fractures and progressive bone deformities, including long bone bowing and scoliosis. Multidisciplinary management improves quality of life for patients with osteogenesis imperfecta. It consists of physical therapy, medical treatment and orthopaedic surgery as necessary. Medical treatment consists of bone-remodelling drug therapy. Bisphosphonates are widely used in the treatment of moderate to severe osteogenesis imperfecta, from infancy to adulthood. Other more recent drug therapies include teriparatide and denosumab. All these therapies target the symptoms and have effects on the mechanical properties of bone due to modification of bone remodelling, therefore influencing skeletal outcome and orthopaedic surgery. Innovative therapies, such as progenitor and mesenchymal stem cell transplantation, targeting the specific altered pathway rather than the symptoms, are in the process of development.


Possible Inhibitory Effect of Erythropoiesis-Stimulating Agents at the Predialysis Stage on Early-Phase Coronary Events after Hemodialysis Initiation.
We examined whether the use of erythropoiesis-stimulating agents (ESAs) to correct anemia at the predialysis stage could inhibit early-phase coronary events after hemodialysis initiation. We enrolled 242 patients with chronic kidney disease who had received continued medical treatments and initiated maintenance hemodialysis from 1 September 2000 to 31 December 2014 at Toujinkai Hospital. Patients with a previous history of blood transfusion or any cardiovascular events or interventions were excluded. The coronary events were followed for 1 year after initiation of hemodialysis. Coronary events occurred in 51 of 242 patients: 10 patients had acute coronary syndrome [9 with percutaneous coronary intervention (PCI), 1 without intervention], and 41 had elective coronary revascularization (38 PCI and 3 coronary artery bypass graft). ESA was administered in 118 of 242 patients (48.8%). In stepwise logistic analysis, coronary events were positively associated with nonuse of ESA at the predialysis stage (odds ratio 2.66, p = 0.005) and diabetes mellitus (odds ratio 5.33, p < 0.001). When dividing the patients into 4 subgroups by blood hemoglobin (Hb) level (8.5 g/dl) and the use/nonuse of ESA, coronary event-free survival rates were higher (p = 0.005) in those with Hb ≥8.5 g/dl, ESA+ (86.6%, n = 82) and tended to be higher (p = 0.055) in those with Hb <8.5 g/dl, ESA+ (86.1%, n = 36) than in patients with Hb <8.5 g/dl, ESA- (68.6%, n = 86) in a Kaplan-Meier analysis. The use of ESA to correct anemia at the predialysis stage may inhibit early-phase coronary events after hemodialysis initiation.


Structures of human MST3 kinase in complex with adenine, ADP and Mn2+.
The MST family is a subclass of mammalian serine/threonine kinases that are related to the yeast sterile-20 protein and are implicated in regulating cell growth and transformation. The MST3 protein contains a 300-residue catalytic domain and a 130-residue regulatory domain, which can be cleaved by caspase and activated by autophosphorylation, promoting apoptosis. Here, five crystal structures of the catalytic domain of MST3 are presented, including a complex with ADP and manganese, a unique cofactor preferred by the enzyme, and a complex with adenine. Similar to other protein kinases, the catalytic domain of MST3 folds into two lobes: the smaller N lobe forms the nucleotide-binding site and the larger C lobe recognizes the polypeptide substrate. The bound ADP and Mn(2+) ions are covered by a glycine-rich loop and held in place by Asn149 and Asp162. A different orientation was observed for the ligand in the MST3-adenine complex. In the activation loop, the side chain of Thr178 is phosphorylated and is sandwiched by Arg143 and Arg176. Comparison of this structure with other similar kinase structures shows a 180 degrees rotation of the loop, leading to activation of the enzyme. The well defined protein-ligand interactions also provide useful information for the design of potent inhibitors.


Vaccine prevention of meningococcal disease, coming soon?
The past century has seen the use of a number of vaccines for prevention and control of meningococcal disease with varied success. The use of polysaccharide vaccines for the control of outbreaks of serogroup C infections in teenagers and young adults and epidemic serogroup A disease has been established for 30 years and an effective protein-polysaccharide conjugate vaccine against serogroup C was introduced into the UK infant immunisation schedule in 2000. The next generation of these glycoconjugate vaccines will be on the shelf soon, eventually offering the prospect of eradication of serogroups A, C, Y and W135 through routine infant immunisation. Despite these exciting prospects, serogroup B meningococci still account for a majority of infections in industrialised nations but development of safe, immunogenic and effective serogroup B meningococcal vaccines has been an elusive goal. Outer membrane vesicle vaccines for B disease are already used in some countries, and will likely be used more widely in the next few years, but efficacy for endemic disease in children has so far been disappointing. However, the innovations arising from the availability of the meningococcal genome sequence, public and scientific interest in the disease and recent pharmaceutical company investment in development of serogroup B vaccines may have started the countdown to the end of meningococcal infection in children.


Training programs within global networks: lessons learned in the Fogarty International Clinical Research Scholars and Fellows Program.
The Fogarty International Clinical Research Scholars and Fellows Support Center at Vanderbilt describes administrative lessons learned from the management of 436 scholars (American students or host country junior trainees) and 122 post-doctoral fellows (Americans or host country nationals). Trainees spent 10-11 months working on mentored research projects at 61 well-vetted sites in 27 low- or middle-income host countries (LMICs) with strong US partners. Economies of scale, strong centralized information exchange, and effective standardized operations linking US institutions with LMIC field sites were achieved in a program that minimized administrative overhead. Advantages and drawbacks of this approach are presented and discussed. Training of a new generation of global research leaders is greatly facilitated by an overseas mentored research experience that is administratively streamlined to optimize the use of resources for training, research, and capacity building.


Use of metformin in the treatment of polycystic ovary syndrome.
Metformin is quite an old drug, but it is optimal for the control of glycemia in Type 2 diabetes. It was reported, 15 years ago, that insulin resistance was abnormally high in most polycystic ovary syndrome (PCOS) patients. Starting from that moment, increasing numbers of studies were performed to demonstrate the efficacy of metformin in controlling and/or modulating several aspects of PCOS, which is the most common cause of menstrual irregularity, inesthetisms and infertility. Metformin induces higher glucose uptake, thus inducing a lower synthesis/secretion of insulin. Such an effect permits the possible restoration of the normal biological functions that are severely affected by the compensatory hyperinsulinemia reactive to the increased peripheral insulin resistance. These are the basis of the many positive effects of this drug, such as the restoration of menstrual cyclicity, ovulatory cycles and fertility, because abnormal insulin levels affect the hypothalamus-pituitary-ovarian function, as well as the use of glucose in peripheral tissues. Metformin improves the impairments typically observed in hyperinsulinemic PCOS patients, reducing the possible evolution towards metabolic syndrome and Type 2 diabetes; and when pregnancy occurs, it consistently reduces the risk of gestational diabetes, eclampsia and hypertension. PCOS seems to be the perfect physiopathological condition that might have higher benefits from metformin administration, obviously after Type 2 diabetes. This review focuses on the many aspects of PCOS and on the possible issues of this disease for which metformin might be a putative optimal treatment.


Modeling and quantifying frequency-dependent fitness in microbial populations with cross-feeding interactions.
Coexistence of two or more populations by frequency-dependent selection is common in nature, and it often arises even in well-mixed experiments with microbes. If ecology is to be incorporated into models of population genetics, then it is important to represent accurately the functional form of frequency-dependent interactions. However, measuring this functional form is problematic for traditional fitness assays, which assume a constant fitness difference between competitors over the course of an assay. Here, we present a theoretical framework for measuring the functional form of frequency-dependent fitness by accounting for changes in abundance and relative fitness during a competition assay. Using two examples of ecological coexistence that arose in a long-term evolution experiment with Escherichia coli, we illustrate accurate quantification of the functional form of frequency-dependent relative fitness. Using a Monod-type model of growth dynamics, we show that two ecotypes in a typical cross-feeding interaction-such as when one bacterial population uses a byproduct generated by another-yields relative fitness that is linear with relative frequency.


Spectral-domain spectrally-encoded endoscopy.
Spectrally-encoded miniature endoscopy uses a single optical fiber and wavelength division multiplexing to obtain macroscopic images through miniature, flexible probes. In turn, it has the potential to enable two- and three-dimensional imaging within the body at locations that are currently difficult to access with conventional endoscopes. Here we present a novel detection scheme for spectrally-encoded endoscopy using spectral-domain interferometry. Compared to previous time-domain configurations, this new detection method results in greater than 1000-fold increase in sensitivity (77 dB), a 6-fold increase in imaging speed (30 volumes per second), and a 2-fold increase in depth range (2.8 mm). We demonstrate spectrally-encoded, spectral-domain detection by conducting video-rate, three-dimensional imaging in a variety of specimens, including the paws of a mouse embryo and excised human ear bones. Our results show that this new technology enables video rate spectrally-encoded endoscopy and will therefore be useful for a variety of minimally invasive medical applications.