Paradigm change and uncertainty about funding of public health research: social and scientific implications.
Since the 1970s two fundamental shifts have occurred in health research funding: a reduction in the buying power of research dollars, and an increase in the competition for resources. Most fields have also seen a decrease in the dollars available for research. Pressures for justifying the relevance of research activities have become increasingly pragmatic. The thesis of this paper is that scientific creativity and innovation are compromised by the highly uncertain and competitive funding environment of contemporary health research. This is largely because criteria of scientific excellence predicated on an investigation's presumed future impact support the status quo of methods and subject matter in funded research. Extraordinary rationality among scientists seeking and allocating resources promotes the survival of the existing system over time, yet inhibits progressive development through the transformation of conceptual models. Therefore, despite a growing unrest about the way research on population health is conducted, new conceptions of the relationship between theory and methods have been slow to emerge. Amelioration of a disjunction between the institutionalized rules governing science and the culturally sanctioned goals of science requires commitment to a dialectic between orthodoxy and dissent.


Transcranial Doppler in spontaneous attacks of migraine.
Our aim in this study was to compare headache-free and spontaneous migraine measurements of blood flow velocity and the pulsatility index in the anterior cerebral artery, middle cerebral artery, and posterior cerebral artery. Thirty-one patients (nine having experienced migraine with aura and 22 migraine without aura) were studied in headache-free periods and during spontaneous migraine attacks with transcranial Doppler ultrasonography. During attacks of migraine with aura, blood flow velocities (particularly the diastolic velocity [p = 0.05]) were reduced while the pulsatility index increased (p = 0.05), whereas a generalized increase in diastolic velocity (p less than 0.02) and a decrease in the pulsatility index (p = 0.05) were observed during attacks of migraine without aura. Significant variations of blood pressure and heart rate were never found. These findings are consistent with constriction of resistance vessels in migraine with aura and dilatation of the vessels in migraine without aura. This disparity could be due to a difference between the two migraine types or could be related to the fact that in this study the time interval between headache onset and transcranial Doppler was shorter in the migraine-with-aura group. The latter explanation would apply if, in fact, both types of migraine evolve from hypoperfusion to hyperperfusion during their time course, although perhaps with a difference in intensity.


Dispelling the disability stereotype: embracing a universalistic perspective of disablement.
The notion of universalism was introduced to me during my first year of PhD studies in Rehabilitation Science. During a class discussion, we debated the merits of two theoretical perspectives that offered contradicting views as to the most effective means to facilitating a shift in societal perceptions of disability. As exemplified by the World Health Organization's current model of health, the International Classification of Functioning, Disability and Health (ICF), there has been a shift from a minority group analysis towards a universalistic perspective of disablement. This paper introduces readers to the underlying concepts of both minority group analysis and universalism and, in doing so, proposes that universalism is closely aligned with the underlying constructs of occupational therapy. Universalism provides a comprehensive framework that can be utilized by occupational therapists to encourage the development of health and social-related policies that promote inclusiveness, yet still the respect the differences that exist among individuals. PRACTICE IMPLICATIONS By improving their familiarity with such theories, occupational therapists may be better positioned to contribute to policy development within their respective treatment and/or community settings.


Perceptions of romantic partners' responsiveness during a period of stressful uncertainty.
Whether awaiting biopsy results, a grade on a midterm, or a decision from a journal editor, people feel distressed as they wait for uncertain news. In the present study, we investigated how people's perceptions of their romantic partner, specifically their partner's responsiveness to their support needs, corresponds with key aspects of the waiting experience. In a longitudinal study of 184 law students awaiting their result on the California bar exam, we examined changes in perceived responsiveness over time and associations between perceived responsiveness and expectation management strategies, health, and well-being. Results revealed temporal patterns in perceived responsiveness, with the greatest responsiveness perceived at the start and end of the wait. Perceived responsiveness was also intertwined with efforts to manage one's expectations while awaiting uncertain news and was associated with more positive emotions, better subjective coping, and greater self-reported sleep quality during the wait. (PsycINFO Database Record


Moderators and mediators of pedometer use and step count increase in the "10,000 Steps Ghent" intervention.
The European pedometer-based "10,000 Steps Ghent" whole community intervention for 228,000 residents was found to be effective in increasing step counts by an average of 896 steps/day in a sub-sample of adults. The present study aimed to examine the characteristics of intervention participants (n = 438) who (1) used a pedometer and (2) increased their step counts. Additionally, the third aim was to examine the mediational effect of pedometer use on step count change. The study sample consisted of 438 adults (207 male, mean age 49.8 (13.1) years). Binary logistic regressions were used to examine whether individual characteristics (gender, age, educational level, employment status, self-reported health condition, baseline step counts, baseline sitting time, baseline transport-related PA) and intervention exposure variables (having heard/seen a PA promotion message, being aware of the PA guidelines, and knowing about "10,000 Steps Ghent") were associated with (1) pedometer use and (2) a step count increase of 896 steps/day or more. Using pooled data (n = 864) from the intervention and comparison participants, a mediation analysis was conducted to see if the change in step counts was mediated by pedometer use. Age (49 years or more: OR = 3.19, p < 0.005), awareness of a PA promotion message (OR = 2.62, p < 0.01) and awareness of "10,000 Steps Ghent" (OR = 2.11, p < 0.05) were significantly associated with pedometer use. Participants with a college or university degree (OR = 1.55, p < 0.05) and those who used a pedometer (OR = 2.06, p < 0.05) were more likely to increase their steps by 896 steps/day or more. This increase was less likely among those with baseline step counts above 10,000 steps/day (OR = 0.38, p < 0.001). The mediation analysis revealed that pedometer use partly mediated step count change. Pedometer use was more likely in older participants and in those who were aware of the "10,000 Steps" campaign. Increasing step counts was more likely among those with higher education, baseline step counts below 10,000 steps/day and those who used a pedometer. Pedometer use only partly mediated the intervention effect on step counts.


Collective credit allocation in science.
Collaboration among researchers is an essential component of the modern scientific enterprise, playing a particularly important role in multidisciplinary research. However, we continue to wrestle with allocating credit to the coauthors of publications with multiple authors, because the relative contribution of each author is difficult to determine. At the same time, the scientific community runs an informal field-dependent credit allocation process that assigns credit in a collective fashion to each work. Here we develop a credit allocation algorithm that captures the coauthors' contribution to a publication as perceived by the scientific community, reproducing the informal collective credit allocation of science. We validate the method by identifying the authors of Nobel-winning papers that are credited for the discovery, independent of their positions in the author list. The method can also compare the relative impact of researchers working in the same field, even if they did not publish together. The ability to accurately measure the relative credit of researchers could affect many aspects of credit allocation in science, potentially impacting hiring, funding, and promotion decisions.


Vascular Endothelial Growth Factor A serum levels and common gene polymorphisms in generalized periodontitis affected patients.
To evaluate and compare the associations of VEGFA serum levels and SNPs (rs1570360, rs699947, rs3025033, and rs2146323) with periodontitis in study participants grouped by gender. The study enrolled 261 patients with periodontitis and 441 healthy controls as a reference group. Patients underwent periodontal examination and radiographic analysis to confirm the periodontitis diagnosis. Blood samples were collected, and the DNA salting-out method was used for DNA extraction from peripheral venous blood. Genotyping of VEGFA (rs1570360, rs699947, rs3025033, and rs2146323) was performed using real-time polymerase chain reaction (RT-PCR) and serum level analysis was done for 80 individuals - 40 periodontitis-affected patients and 40 reference group subjects. The analysis of VEGFA (rs1570360, rs699947, rs3025033, and rs2146323) showed that the rs3025033 GG genotype was less frequent in the periodontitis group than in the reference group (1.6% vs. 5.7%,p = 0.008). VEGFA serum levels were not statistically significantly different between periodontitis patients and reference group subjects (554.29 (522.38) ng/ml vs. 581.32 (348.16) ng/ml, p = 0.786). Individuals carrying rs1570360, rs699947, rs3025033, and rs2146323 haplotype A-A-G-A had decreased risks of periodontitis, while rare haplotype of VEGFA (rs1570360, rs699947, rs3025033, and rs2146323) was associated with increased odds of periodontitis (OR= 0.42; 95% CI: 0.20-0.85; p < 0.017; OR= 4.08; 95% CI: 1.86-8.94; p < 0.0001, respectively). The rs3025033 GG genotype and the rs1570360, rs699947, rs3025033, and rs2146323 A-A-G-A haplotypes may play a protective role in the development of periodontitis, but a less common haplotype of the same VEGFA polymorphism may be associated with the risk of developing periodontitis.


Effect of the G-308A polymorphism of the tumor necrosis factor (TNF)-alpha gene promoter site on plasma levels of TNF-alpha and C-reactive protein in smokers: a cross-sectional study.
Plasma levels of tumor necrosis factor (TNF)-alpha and of C-reactive protein (CRP) are elevated in smokers. Previous studies failed to show an association between the G-308A polymorphism in the promoter region of the TNF-alpha gene and coronary artery disease (CAD). We investigated whether smoking would interact with the TNF-alpha G-308A polymorphism in determining plasma levels of TNF-alpha and CRP. Study participants with a complete data set in terms of smoking and the TNF-alpha G-308A polymorphism were 300 middle-aged male and female industrial employees. After excluding 24 irregular smokers, analyses were performed on 198 "non-smokers" (life-long non-smokers or subjects who quit smoking >6 months ago) as compared to 78 "regular smokers" (subjects currently smoking >3 cigarettes/day). All subjects had a fasting morning blood draw to measure plasma levels of TNF-alpha and CRP by high-sensitive enzyme-linked immunosorbent assays. The cardiovascular risk factor adjusted analysis regressing log-transformed CRP levels against smoking status, genotype, and smoking-status-genotype interaction revealed a significant main effect for smoking status (F1,250 = 5.67, p = .018) but not for genotype (F1,250 = 0.33, p = .57). The interaction-term between genotype and smoking status was not significant (F1,250 = 0.09, p = .76). The fully adjusted model with plasma TNF-alpha failed to show significant main effects for smoking and genotype, as well as for the smoking-status-genotype interaction. The findings suggest that the TNF-alpha G-308A polymorphism does not mediate the effect of smoking on plasma CRP levels. It remains to be seen whether other genetic polymorphisms along the inflammatory pathway may modulate vascular risk in smokers.


Size scaling in collective cell growth.
Size is a fundamental feature of living entities and is intimately tied to their function. Scaling laws, which can be traced to D'Arcy Thompson and Julian Huxley, have emerged as a powerful tool for studying regulation of the growth dynamics of organisms and their constituent parts. Yet, throughout the 20th century, as scaling laws were established for single cells, quantitative studies of the coordinated growth of multicellular structures have lagged, largely owing to technical challenges associated with imaging and image processing. Here, we present a supervised learning approach for quantifying the growth dynamics of germline cysts during oogenesis. Our analysis uncovers growth patterns induced by the groupwise developmental dynamics among connected cells, and differential growth rates of their organelles. We also identify inter-organelle volumetric scaling laws, finding that nurse cell growth is linear over several orders of magnitude. Our approach leverages the ever-increasing quantity and quality of imaging data, and is readily amenable for studies of collective cell growth in other developmental contexts, including early mammalian embryogenesis and germline development.


Adenovirus-mediated expression of p53 or p21 in a papillary serous endometrial carcinoma cell line (SPEC-2) results in both growth inhibition and apoptotic cell death: potential application of gene therapy to endometrial cancer.
Papillary serous endometrial carcinoma is an aggressive tumor characterized by late-stage presentation, i.p. spread, and poor prognosis. It is histologically similar to serous papillary carcinoma of the ovary. Preclinical studies have shown that adenovirus-mediated expression of p53 in ovarian cancer cell lines causes growth inhibition and apoptosis in vitro and in vivo. Such studies provide the rationale for Phase I Adp53 gene therapy clinical trials in ovarian cancer. In the present study, we compared the efficacy of adenoviral vectors containing p53 (Adp53) or p21 (Adp21) in a papillary serous endometrial tumor cell line (SPEC-2) that contains mutated p53. Growth assays revealed that both Adp53 and Adp21 were efficacious in decreasing cell proliferation as assessed by anchorage-dependent and anchorage-independent growth assays. However, as compared with Adp53, the effects of Adp21 tended to be more transient and less marked. Strikingly, Adp21, but not Adp53, induced a G1 arrest in SPEC-2 endometrial adenocarcinoma cells. In contrast, as assessed by induction of hypodiploid peaks, free DNA ends detected by a terminal deoxynucleotidyl transferase-based assay, and annexin V positivity, p53 was more effective than p21 in inducing cell death by apoptosis. Compatible with the more efficient induction of apoptosis, Adp53, but not Adp21, induced a marked increase in expression of the preapoptotic molecule BAX without a concomitant change in expression of the antiapoptotic mediator Bcl-2. The differential effects of Adp53 and Adp21 on cell cycle progression and apoptosis may be related to the reversibility of p21-induced cell cycle arrest and the irreversibility of p53-induced apoptosis. Thus, at least in the papillary serous endometrial carcinoma cell line SPEC-2, Adp53 may be more effective than Adp21 as a gene therapeutic. Nevertheless, these preclinical studies suggest that papillary serous endometrial carcinoma is a potential target for p53- or p21-mediated gene therapy.


Disparities in self-monitoring of blood glucose among low-income ethnic minority populations with diabetes, United States.
In adults with insulin-treated diabetes, self-monitoring of blood glucose (SMBG) rates may be lower in minority or low-income populations, but the effect of income on racial/ethnic differences in SMBG is unknown. We assessed whether racial/ethnic differences in SMBG vary by income among adults with insulin-treated diabetes by using Behavioral Risk Factor Surveillance System data from 2000 through 2004. We measured the prevalence of SMBG at least once per day among 16,630 adults aged > or = 19 years with insulin-treated diabetes. At incomes > or = $20,000, Hispanics and non-Hispanic Blacks reported similar but lower SMBG rates than did non-Hispanic Whites (78%, 77%, 85%; P < or = .01). However, among those with income < $20,000, Hispanics performed SMBG substantially less than did Blacks or Whites (65%, 79%, 85%; P < or = .01). Racial/ ethnic differences in SMBC persisted after adjustment for age, sex, education, health insurance, health status, survey period, and diabetes measures. Receipt of diabetes education varied significantly by race/ethnicity in the income < $20,000 group only (Hispanics 49%, Blacks 64%, Whites 62%; P < .001). Low-income Hispanics with limited English proficiency had lower SMBG and diabetes education rates than did those with English proficiency (61% vs 79% and 44% vs 58%, respectively). Among US adults with insulin-treated diabetes, Hispanics and Blacks performed daily SMBG less frequently than did Whites. Stratification by income revealed a disparity gradient in the income < $20,000 group: SMBG rates decreased from Whites to Blacks to Hispanics. Low-income Hispanics with limited English proficiency are at greater risk for reduced SMBG than are those proficient in English.


Review and publication of protocol submissions to Trials - what have we learned in 10 years?
Trials has 10 years of experience in providing open access publication of protocols for randomised controlled trials. In this editorial, the senior editors and editors-in-chief of Trials discuss editorial issues regarding managing trial protocol submissions, including the content and format of the protocol, timing of submission, approaches to tracking protocol amendments, and the purpose of peer reviewing a protocol submission. With the clarification and guidance provided, we hope we can make the process of publishing trial protocols more efficient and useful to trial investigators and readers.


Temporal relationship between 18F-sodium fluoride uptake in the abdominal aorta and evolution of CT-verified vascular calcification.
Fluoride-18 sodium fluoride (18F-NaF) localizes in microcalcifications in atheroma. The microcalcifications may aggregate, passing the resolution threshold to visualize on computed tomography (CT). We evaluated serial NaF positron emission tomography (PET)-CT scans to determine the temporal relationship between vascular NaF uptake and CT evident calcification in the abdominal aorta. Prostate cancer patients who had at least 3 NaF PET-CT scans over at least 1.5 years were retrospectively enrolled. Regions of interest were traced in the abdominal aorta on both PET and CT images, excluding skeletal NaF activity. The maximum standardized uptake value (SUVmax) of NaF and the density and volume of calcium (exceeding 130 HU) were summed and divided by the number of slices to produce the SUVmax/slice and the mm3·slice-1 of calcium. Of 437 patients, 45 patients met criteria. NaF uptake waxed and waned between scans, while the calcium volume plateaued or increased over time. NaF uptake correlated with calcium volume on the baseline scan (P = .60, < .0001†) and calcium volume increment, especially from 1.0 to 1.5 years (r = .79, P < .0001†). Patients with persistently high NaF uptake showed a higher calcium volume increment (0-1.5 years) than patients with low or transiently high NaF uptake. Abdominal aortic NaF uptake varied over time. NaF uptake on the baseline scans and high NaF uptake on the serial scans preceded an increase in calcium volume, especially by 1.0-1.5 years. Persistently high NaF uptake was associated with a greater increment in calcium volume than patients with transiently elevated or persistently low fluoride uptake.


Major dietary patterns and carotid intima-media thickness in Bangladesh.
Carotid intima-media thickness (IMT) is a validated surrogate marker of preclinical atherosclerosis and is predictive of cardiovascular morbidity and mortality. Research on the association between IMT and diet, however, is lacking, especially in low-income countries or low-BMI populations. Cross-sectional analysis. Dietary intakes were measured using a validated, thirty-nine-item FFQ at baseline cohort recruitment. IMT measurements were obtained from 2010-2011. Rural Bangladesh. Participants (n 1149) randomly selected from the Health Effects of Arsenic Longitudinal Study, an ongoing, population-based, prospective cohort study established in 2000. Average age at IMT measurement was 45·5 years. Principal component analysis of reported food items yielded a 'balanced' diet, an 'animal protein' diet and a 'gourd and root vegetable' diet. We observed a positive association between the gourd/root vegetable diet and IMT, as each 1 sd increase in pattern adherence was related to a difference of 7·74 (95 % CI 2·86, 12·62) μm in IMT (P<0·01), controlling for age, sex, total energy intake, smoking status, BMI, systolic blood pressure and diabetes mellitus diagnoses. The balanced pattern was associated with lower IMT (-4·95 (95 % CI -9·78, -0·11) μm for each 1sd increase of adherence; P=0·045). A gourd/root vegetable diet in this Bangladeshi population positively correlated with carotid IMT, while a balanced diet was associated with decreased IMT.


Diabetes type II: a risk factor for depression-Parkinson-Alzheimer?
There is ample evidence that impairments in the hypothalamic-pituitary-adrenal (HPA) axis are of etiopathobiochemical importance in a subgroup of patients with "depression", causing hypercortisolaemia as major metabolic effect. Chronic hypercortisolaemia causes insulin resistance. Therefore, it is not surprising that epidemiological studies demonstrate an association of "depression" with diabetes type II and vice versa. Chronic stress and hypercortisolaemia are conditions, which have been suggested to be causal for Alzheimer's disease (AD) as brain insulin resistance is associated with β-Amyloid-accumulation and hyperphosphorylation of tau-protein. Depression is one of the significant symptomatology preceding AD. It is however, not known whether "depression" associated with hypercortisolaemia is the subgroup at risk for AD. In contrast to a subgroup of "depression" and to AD, in Parkinson's disease (PD) there is only weak evidence for an association with diabetes type II and insulin resistance. As "depression" is preceding PD in up to half of such patients, it remains to be elucidated whether this is a subgroup of depressed patients, which is not associated with disturbances of the HPA axis and hypercortisolaemia. Improved clinical and biochemical/molecular knowledge about "depression" associated with AD and PD in comparison to "pure" depression might lead to improved therapeutic strategies and even drug development focusing subtypes of "depression".


Quantitative planar rest-redistribution 201Tl imaging in detection of myocardial viability and prediction of improvement in left ventricular function after coronary bypass surgery in patients with severely depressed left ventricular function.
Although many patients with multivessel coronary artery disease (CAD) and severely depressed left ventricular (LV) function will benefit from coronary artery bypass graft surgery (CABG), surgeons may be reluctant to perform CABG on these patients without evidence of myocardial viability in regions of severe asynergy. We hypothesized that quantitative planar rest-redistribution 201Tl imaging would identify viable myocardium and predict improved regional and global function after revascularization in patients with depressed LV function and CAD. Twenty-one patients (mean LV ejection fraction, 0.27 +/- 0.05) were studied. Regional and global LV functions were evaluated before and 8 weeks after CABG with radionuclide ventriculography. Segments were prospectively classified as showing normal, mildly reduced, or severely reduced viability on the basis of quantitative analysis of defect severity and redistribution on planar resting 201Tl imaging. By 201Tl criteria, 90% of hypokinetic segments were classified with normal or mildly reduced viability. Among akinetic or dyskinetic segments, 20% had normal 201Tl uptake, 53% had mildly reduced viability, and only 27% had severely reduced viability. 201Tl viability criteria identified segments that improved function after CABG. Sixty-two percent of severely asynergic segments with normal viability and 54% with mildly reduced viability improved function after surgery, but only 23% with severely reduced viability improved function (p = 0.002). When only adequately revascularized segments were considered, the predictive value of a positive preoperative viability scan for functional improvement was 73%. The greatest improvement in global LV function after CABG occurred in patients with the greatest number of asynergic segments classified as viable before surgery (p < 0.01). In 10 patients with more than seven viable, asynergic segments, mean LV ejection fraction increased significantly after CABG (0.29 +/- 0.07 to 0.41 +/- 0.11, p = 0.002). In 11 patients with seven or fewer viable, asynergic segments, mean LV ejection fraction remained unchanged after revascularization (0.27 +/- 0.05 to 0.30 +/- 0.08, p = NS). In patients with CAD and severely depressed LV function, preoperative quantitative planar rest-redistribution. 201Tl imaging identifies viability in many asynergic myocardial segments, and these segments frequently improve function after CABG. The presence of numerous asynergic but viable myocardial segments before surgery correlated significantly with improvement in global LV function after bypass surgery.


Outcomes of BRAF V600E Pediatric Gliomas Treated With Targeted BRAF Inhibition.
Children with pediatric gliomas harboring a BRAF V600E mutation have poor outcomes with current chemoradiotherapy strategies. Our aim was to study the role of targeted BRAF inhibition in these tumors. We collected clinical, imaging, molecular, and outcome information from patients with BRAF V600E-mutated glioma treated with BRAF inhibition across 29 centers from multiple countries. Sixty-seven patients were treated with BRAF inhibition (pediatric low-grade gliomas [PLGGs], n = 56; pediatric high-grade gliomas [PHGGs], n = 11) for up to 5.6 years. Objective responses were observed in 80% of PLGGs, compared with 28% observed with conventional chemotherapy (P < .001). These responses were rapid (median, 4 months) and sustained in 86% of tumors up to 5 years while receiving therapy. After discontinuation of BRAF inhibition, 76.5% (13 of 17) of patients with PLGG experienced rapid progression (median, 2.3 months). However, upon rechallenge with BRAF inhibition, 90% achieved an objective response. Poor prognostic factors in conventional therapies, such as concomitant homozygous deletion of CDKN2A, were not associated with lack of response to BRAF inhibition. In contrast, only 36% of those with PHGG responded to BRAF inhibition, with all but one tumor progressing within 18 months. In PLGG, responses translated to 3-year progression-free survival of 49.6% (95% CI, 35.3% to 69.5%) versus 29.8% (95% CI, 20% to 44.4%) for BRAF inhibition versus chemotherapy, respectively (P = .02). Use of BRAF inhibition results in robust and durable responses in BRAF V600E-mutated PLGG. Prospective studies are required to determine long-term survival and functional outcomes with BRAF inhibitor therapy in childhood gliomas.


Kinetics and strain specificity of rhizosphere and endophytic colonization by enteric bacteria on seedlings of Medicago sativa and Medicago truncatula.
The presence of human-pathogenic, enteric bacteria on the surface and in the interior of raw produce is a significant health concern. Several aspects of the biology of the interaction between these bacteria and alfalfa (Medicago sativa) seedlings are addressed here. A collection of enteric bacteria associated with alfalfa sprout contaminations, along with Escherichia coli K-12, Salmonella enterica serotype Typhimurium strain ATCC 14028, and an endophyte of maize, Klebsiella pneumoniae 342, were labeled with green fluorescent protein, and their abilities to colonize the rhizosphere and the interior of the plant were compared. These strains differed widely in their endophytic colonization abilities, with K. pneumoniae 342 and E. coli K-12 being the best and worst colonizers, respectively. The abilities of the pathogens were between those of K. pneumoniae 342 and E. coli K-12. All Salmonella bacteria colonized the interiors of the seedlings in high numbers with an inoculum of 10(2) CFU, although infection characteristics were different for each strain. For most strains, a strong correlation between endophytic colonization and rhizosphere colonization was observed. These results show significant strain specificity for plant entry by these strains. Significant colonization of lateral root cracks was observed, suggesting that this may be the site of entry into the plant for these bacteria. At low inoculum levels, a symbiosis mutant of Medicago truncatula, dmi1, was colonized in higher numbers on the rhizosphere and in the interior by a Salmonella endophyte than was the wild-type host. Endophytic entry of M. truncatula appears to occur by a mechanism independent of the symbiotic infections by Sinorhizobium meliloti or mycorrhizal fungi.


Simple and scalable electrochemical synthesis of 2,1-benzisoxazoles and quinoline N-oxides.
Cathodic reduction of the nitro moiety and subsequent intramolecular cyclization affords different substituted 2,1-benzisoxazoles and quinoline N-oxides. This methodology allows the synthesis of two different types of heterocycles from common simple starting materials, using electrons as a sole reagent for this transformation. The electrolysis can be conducted in a very simple undivided electrolysis cell under constant current conditions. This permits working on a larger scale compared to other electrochemical methodologies and represents a significant advantage.


Evaluation of excited state bond weakening for ammonia synthesis from a manganese nitride: stepwise proton coupled electron transfer is preferred over hydrogen atom transfer.
Concepts for the thermodynamically challenging synthesis of weak N-H bonds by photoinduced proton coupled electron transfer are explored. Upon irradiation with blue light, ammonia synthesis was achieved from the manganese nitride (tBuSalen)MnN (tBuSalen = (S,S)-(+)-N,N'-bis(3,5-di-tert-butylsalicylidene)-1,2-cyclohexanediamine) in the presence of 9,10-dihydroacridine and a ruthenium photocatalyst in iPrOH solution. Although in one case the ruthenium complex bears a remote N-H bond that weakens to 41 kcal mol-1 upon irradiation, control experiments with the N-methylated analog demonstrate the ruthenium complex serves as a photoreductant rather than hydrogen-atom transfer catalyst in aprotic solvents. Luminescence quenching experiments support a ruthenium(ii)/(iii) cycle rather than a ruthenium(i)/(ii) alternative. Identification of the manganese complex following ammonia synthesis was also accomplished.


Expression of genes encoding multi-transmembrane proteins in specific primate taste cell populations.
Using fungiform (FG) and circumvallate (CV) taste buds isolated by laser capture microdissection and analyzed using gene arrays, we previously constructed a comprehensive database of gene expression in primates, which revealed over 2,300 taste bud-associated genes. Bioinformatics analyses identified hundreds of genes predicted to encode multi-transmembrane domain proteins with no previous association with taste function. A first step in elucidating the roles these gene products play in gustation is to identify the specific taste cell types in which they are expressed. Using double label in situ hybridization analyses, we identified seven new genes expressed in specific taste cell types, including sweet, bitter, and umami cells (TRPM5-positive), sour cells (PKD2L1-positive), as well as other taste cell populations. Transmembrane protein 44 (TMEM44), a protein with seven predicted transmembrane domains with no homology to GPCRs, is expressed in a TRPM5-negative and PKD2L1-negative population that is enriched in the bottom portion of taste buds and may represent developmentally immature taste cells. Calcium homeostasis modulator 1 (CALHM1), a component of a novel calcium channel, along with family members CALHM2 and CALHM3; multiple C2 domains; transmembrane 1 (MCTP1), a calcium-binding transmembrane protein; and anoctamin 7 (ANO7), a member of the recently identified calcium-gated chloride channel family, are all expressed in TRPM5 cells. These proteins may modulate and effect calcium signalling stemming from sweet, bitter, and umami receptor activation. Synaptic vesicle glycoprotein 2B (SV2B), a regulator of synaptic vesicle exocytosis, is expressed in PKD2L1 cells, suggesting that this taste cell population transmits tastant information to gustatory afferent nerve fibers via exocytic neurotransmitter release. Identification of genes encoding multi-transmembrane domain proteins expressed in primate taste buds provides new insights into the processes of taste cell development, signal transduction, and information coding. Discrete taste cell populations exhibit highly specific gene expression patterns, supporting a model whereby each mature taste receptor cell is responsible for sensing, transmitting, and coding a specific taste quality.


Superior Memory Reduces 8-year Risk of Mild Cognitive Impairment and Dementia But Not Amyloid β-Associated Cognitive Decline in Older Adults.
To prospectively examine 8-year risk of clinical disease progression to mild cognitive impairment (MCI)/dementia in older adults ≥60 with superior episodic memory (SuperAgers) compared to those cognitively normal for their age (CNFA). Additionally, to determine the extent to which SuperAgers were resilient to the negative effects of elevated amyloid-beta (Aβ+) on cognition. Participants were classified as SuperAgers based on episodic memory performance consistent with younger adults aged 30-44 and no impairment on non-memory tests (n = 179), and were matched with CNFA on age, sex, education, and follow-up time (n = 179). Subdistribution hazard models examined risk of clinical progression to MCI/dementia. Linear mixed models assessed the effect of Aβ on cognition over time. Prevalence of Aβ+ and APOE ε4 was equivalent between SuperAgers and CNFA. SuperAgers had 69%-73% reduced risk of clinical progression to MCI/dementia compared to CNFA (HR: 0.27-0.31, 95% CI: 0.11-0.73, p < .001). Aβ+ was associated with cognitive decline in verbal memory and executive function, regardless of SuperAger/CNFA classification. In the absence of Aβ+, equivalent age-related changes in cognition were observed between SuperAgers and CNFA. SuperAgers displayed resilience against clinical progression to MCI/dementia compared to CNFA despite equivalent risk for Alzheimer's disease (AD); however, SuperAgers had no greater protection from Aβ+ than CNFA. The deleterious effects of Aβ on cognition persist regardless of baseline cognitive ability. Thus, superior cognitive performance does not reflect resistance against the neuropathological processes associated with AD, and the observed resilience for SuperAgers may instead reflect neuropsychological criteria for cognitive impairment.


Statins Reverse Postpartum Cardiovascular Dysfunction in a Rat Model of Preeclampsia.
Preeclampsia is associated with increased cardiovascular long-term risk; however, the underlying functional and structural mechanisms are unknown. We investigated maternal cardiac alterations after preeclampsia. Female rats harboring the human angiotensinogen gene [TGR(hAogen)L1623] develop a preeclamptic phenotype with hypertension and albuminuria during pregnancy when mated with male rats bearing the human renin gene [TGR(hRen)L10J] but behave physiologically normal before and after pregnancy. Furthermore, rats were treated with pravastatin. We tested the hypothesis that statins are a potential therapeutic intervention to reduce cardiovascular alterations due to simulated preeclamptic pregnancy. Although hypertension persists for only 8 days in pregnancy, former preeclampsia rats exhibit significant cardiac hypertrophy 28 days after pregnancy observed in both speckle tracking echocardiography and histological staining. In addition, fibrosis and capillary rarefaction was evident. Pravastatin treatment ameliorated the remodeling and improved cardiac output postpartum. Preeclamptic pregnancy induces irreversible structural changes of cardiac hypertrophy and fibrosis, which can be moderated by pravastatin treatment. This pathological cardiac remodeling might be involved in increased cardiovascular risk in later life.


Collagen cross-linking in thin corneas.
Collagen cross-linking (CXL) has become the standard of care for progressive keratoconus, after numerous clinical studies have established its efficacy and safety in suitably selected eyes. The standard protocol is applicable in eyes which have a minimum corneal thickness of 400 μm after epithelial debridement. This prerequisite was stipulated to protect the corneal endothelium and intraocular tissues from the deleterious effect of ultraviolet-A (UVA) radiation. However, patients with keratoconus often present with corneal thickness of less than 400 μm and could have otherwise benefited from this procedure. A few modifications of the standard procedure have been suggested to benefit these patients without a compromise in safety. Transepithelial cross-linking, pachymetry-guided epithelial debridement before cross-linking, and the use of hypoosmolar riboflavin are some of the techniques that have been attempted. Although clinical data is limited at the present time, these techniques are worth considering in patients with thin corneas. Further studies are needed to scientifically establish their efficacy and safety.


Pelvic reconstruction after subtotal sacrectomy for sacral chondrosarcoma using cadaveric and vascularized fibula autograft: Technical note.
A novel method of spinopelvic ring reconstruction after partial sacrectomy for a chondrosarcoma is described. Chondrosarcoma is one of the most common primary malignant bone tumors, and en bloc resection is the mainstay of treatment. Involvement of the pelvis as well as the sacrum and lumbar spine can result in a technically difficult challenge for en bloc resection and for achievement of appropriate load-bearing reconstruction. After en bloc resection in their patient, the authors achieved reconstruction with a rod and screw construct including vascularized fibula graft as the main strut from the lumbar spine to the pelvis. Additionally, a cadaveric allograft strut was used as an adjunct for the pelvic ring. This is similar to a modified Galveston technique with vascularized fibula in place of the Galveston rods. The vascularized fibula provided appropriate biomechanical support, allowing the patient to return to independent ambulation. There was no tumor recurrence; neurological status remained stable; and the allograft construct integrated well and even increased in size on CT scans and radiographs in the course of a follow-up longer than 7 years.


Psychometric properties and cultural adaptation of sleep disturbance measures in Arabic-speaking populations: A systematic review.
The aim of this review was to evaluate the psychometric properties and cross-cultural adaptation of sleep disturbance scales that have been translated into Arabic or originally developed in Arabic, and to identify appropriate scales that can be used in research and clinical practice intended for Arabic-speaking participants. The following databases were searched: CINAHL (2003-2019), MEDLINE (1946-2019), EMBASE (1980-2019), PsycINFO (1806-2019) and Cochrane Library (1806-2019). This review was conducted following PRISMA guidelines. Terwee et al. (J. Clin. Epidemiol., 60, 2007, 34) quality assessment was used to evaluate the psychometric properties of the studies, and cross-cultural adaptation was assessed using criteria from Guillemin, Bombardier, and Beaton (J. Clin. Epidemiol., 46, 1993, 1417). Seven studies met the inclusion criteria, which included four scales: the Epworth Sleepiness Scale, Insomnia Severity Index, Pittsburgh Sleep Quality Index, and Arabic Scale of Insomnia. Cross-cultural adaptations scored between good and poor; psychometric properties information was missing for most scales. The review suggested that Pittsburgh Sleep Quality Index may be a useful scale to measure sleep disturbance, as the scale showed good cultural adaptation and acceptable psychometric properties in an Arabic population. Furthermore, the scales measure seven different aspects of sleep quality. This review provides options to help researchers and clinicians select the most appropriate instrument for their practice. Further psychometric testing and cultural adaptation is required for sleep scales used in Arabic clinical populations to ensure validity and reliability in outcome measurement for research studies.


Huge epithelial nonparasitic splenic cyst: A case report and a review of treatment methods.
Splenic cysts are rare in all age groups and there are a few reports in the world literature. Primary cysts occur most frequently in children and young adults, comprising around 25% of all nonparasitic splenic cysts. Various techniques are suggested for the treatment of splenic cysts. In this case report, a huge epithelial splenic cyst in a 17-year-old female is presented and different treatment methods of splenic cysts are evaluated. A 17-year-old female presented with progressive abdominal mass in left upper quadrant associated with abdominal pain and food intolerance of duration of several months. There was no history of trauma. On physical examination, there was a huge mass located in the upper left side of abdomen. Computerized tomography scan revealed that a large cystic lesion had occupied the spleen with dimensions of 32x21xI5.6 cm. After patient preparation laparotomy was performed and complete cyst excision was done with splenectomy, patient was discharged after 2 days. This is a report of a case of epithelial splenic cyst of the spleen in a 17-year old female. The management of splenic cysts continues to evolve and the optimum treatment of patients with nonparasitic splenic cysts is controversial, as a principle preservation technique of the spleen with minimally invasive methods such as laparoscopy is preferred to splenectomy with the exception of very large cysts and when splenic hilum is involved in cyst wall. However, significant cyst recurrences were encountered with these techniques. Recently open partial splenectomy has been proposed as a safe and effective method in the management of NPSCs it ensures complete cyst removal, lack of cyst recurrence, and preservation of the spleen functions.


Mapping human somatosensory cortex with positron emission tomography.
Positron emission tomography measurements of regional cerebral blood flow were used to detect focal neuronal activation in the first somatosensory cortex (SI) of humans induced by cutaneous vibratory stimulation. Intravenously administered water labeled with oxygen-15 (H2(15)O) was used as a blood flow tracer to obtain five stimulated-state and two resting-state blood flow images in each of eight normal volunteers. Three cutaneous surfaces were tested: lips, fingers, and toes. Intense, highly focal SI responses were seen during all 39 stimulated-state trials. The SI responses from the three stimulation sites were anatomically distinct and formed a medial-to-lateral homonculus in every subject. Response magnitudes (increase in local blood flow) and response locales (expressed as proportionately measured bicommissural stereotaxic coordinates) were highly consistent among subjects and on repeated trials for each subject. These findings suggest that eliciting cerebral blood flow responses by cutaneous vibration provides a safe, rapid, and reproducible tool for locating and assessing the functional status of somatosensory cortex, and offers potential clinical and research utility. This study has established normative values for future applications of this experimental paradigm.


Autoantibodies in patients with gut motility disorders and enteric neuropathy.
Enteric neuropathy with mild inflammation (ganglionitis) has been described in several motility disorders including irritable bowel syndrome (IBS), enteric dysmotility (ED), slow-transit constipation (STC) and chronic intestinal pseudo-obstruction (CIPO). The purpose of this study was to test the hypothesis that autoantibodies directed against specific neural antigens including ion channels may be associated with this finding. Comprehensive routine and immunohistochemical analyses of full-thickness jejunal laparoscopic biopsies were performed on patients fulfilling the international criteria for IBS, ED, STC and CIPO. Patients with ganglionitis had sera screened for specific antibodies to voltage-gated calcium channels (VGCCs) of P/Q- and N-type, voltage-gated potassium channels (VGKCs), glutamic acid decarboxylase (GAD) and neuronal alpha3-AChR by validated immunoprecipitation assays. Thirty-three patients were included in the study. Two of them, both with IBS, were found to have positive antibody titres. One had anti-VGKC antibodies and one had anti-alpha3-AChR antibodies. No antibodies were detected in GAD or VGCCs (case reports presented). A small proportion of patients with inflammatory enteric neuropathy have antibodies directed towards neuronal ion channels. The pathogenic role of such antibodies requires determination but may represent a possible aetiology of severe functional symptoms in these groups of patients.


Prevention and control of tuberculosis in correctional and detention facilities: recommendations from CDC. Endorsed by the Advisory Council for the Elimination of Tuberculosis, the National Commission on Correctional Health Care, and the American Correctional Association.
Tuberculosis (TB) control can be particularly problematic in correctional and detention facilities, in which persons from diverse backgrounds and communities are housed in close proximity for varying periods. This report provides a framework and general guidelines for effective prevention and control of TB in jails, prisons, and other correctional and detention facilities. Recommendations were developed on the basis of published guidelines and a review of the scientific literature. Effective TB-prevention and -control measures in correctional facilities include early identification of persons with TB disease through entry and periodic follow-up screening; successful treatment of TB disease and latent TB infection; appropriate use of airborne precautions (e.g., airborne infection isolation, environmental controls, and respiratory protection); comprehensive discharge planning; and thorough and efficient contact investigation. These measures should be instituted in close collaboration with local or state health department TB-control programs and other key partners. Continuing education of inmates, detainees, and correctional facility staff is necessary to maximize cooperation and participation. To ensure TB-prevention and -control measures are effective, periodic program evaluation should be conducted.


Embelin reduces colitis-associated tumorigenesis through limiting IL-6/STAT3 signaling.
The interleukin-6 (IL-6)/STAT3 signaling regulates survival and proliferation of intestinal epithelial cells and plays an important role in the pathogenesis of inflammatory bowel disease and colorectal cancer. Embelin is a small molecule inhibitor of X-linked inhibitor of apoptosis protein (XIAP), with antioxidant, anti-inflammatory, and antitumor activities. We previously showed that embelin inhibits the growth of colon cancer cells in vitro, and effectively suppresses 1,2-dimethylhydrazine dihydrochloride-induced colon carcinogenesis in mice. Here, we explored the antitumor effects and mechanisms of embelin on colitis-associated cancer (CAC) using the azoxymethane/dextran sulfate sodium (AOM/DSS) model, with a particular focus on whether embelin exerts its effect through the IL-6/STAT3 pathway. We found that embelin significantly reduced incidence and tumor size in CAC-bearing mice. In addition to inhibiting proliferation of tumor epithelial cells, embelin suppressed colonic IL-6 expression and secretion, and subsequently STAT3 activation in vivo. Importantly, in vitro studies have revealed that in colon cancer cells, embelin diminished both the constitutive and IL-6-induced STAT3 activation by stimulating Src homology domain 2-containing protein tyrosine phosphatase (SHP2) activity. Moreover, embelin protected mice from AOM/DSS-induced colitis before tumor development. Embelin decreased IL-1β, IL-17a, and IL-23a expression as well as the number of CD4(+) T cells and macrophages infiltrating the colonic tissues. Thus, our findings demonstrated that embelin suppresses CAC tumorigenesis, and its antitumor effect is partly mediated by limiting IL-6/STAT3 activation and Th17 immune response. Embelin may be a potential agent in the prevention and treatment of CAC.


Parenting practices and child disruptive behavior problems in early elementary school. Conduct Problems Prevention Research Group.
Examined the hypothesis that distinct parenting practices may be associated with type and profile of a child's disruptive behavior problems (e.g., oppositional, aggressive, hyperactive). Parents of 631 behaviorally disruptive children described the extent to which they experienced warm and involved interactions with their children and the extent to which their discipline strategies were inconsistent and punitive and involved spanking and physical aggression. As expected from a developmental perspective, parenting practices that included punitive interactions were associated with elevated rates of all child disruptive behavior problems. Low levels of warm involvement were particularly characteristic of parents of children who showed elevated levels of oppositional behaviors. Physically aggressive parenting was linked more specifically with child aggression. In general, parenting practices contributed more to the prediction of oppositional and aggressive behavior problems than to hyperactive behavior problems, and parenting influences were fairly consistent across ethnic groups and sex.


Actin Dynamics and HIV-1 Entry.
Cortical F-actin plays important roles during HIV-1 infection. A recent paper in Cell highlights the involvement of F-actin in enhancing dendritic cell (DC)-mediated HIV-1 infection of T cells. Such a critical mechanism of HIV-1 trans-enhancement between human DCs and T cells had not been analyzed in molecular detail before this work.


Sensitive and specific single-molecule sequencing of 5-hydroxymethylcytosine.
We describe strand-specific, base-resolution detection of 5-hydroxymethylcytosine (5-hmC) in genomic DNA with single-molecule sensitivity, combining a bioorthogonal, selective chemical labeling method of 5-hmC with single-molecule, real-time (SMRT) DNA sequencing. The chemical labeling not only allows affinity enrichment of 5-hmC-containing DNA fragments but also enhances the kinetic signal of 5-hmC during SMRT sequencing. We applied the approach to sequence 5-hmC in a genomic DNA sample with high confidence.


Studies on application of annatto (Bixa orellena L.) dye formulations in dairy products.
Annatto is often used to add color to dairy products such as butter, cheese, or puddings. In India usage of annatto is restricted to butter and cheese, however there are no tailor made formulations available to obtain standardized colour shades for the products. Hence a study was initiated to develop the appropriate annatto dye formulations and level of application in few dairy products (butter, cheese, paneer, biscuit cream, icing cream). Dye extracted from annatto seeds was used for the preparation of water soluble potassium carbonate formulation (nor-bixin, 11.24 %), oil soluble formulation (1.35 % bixin) and an oil/water soluble propylene glycol formulation (PG formulation, 1.31 % bixin). Lovibond tintometer colour units of the commercial butter, cheese and biscuit cream samples were measured for standardizing the colour concentration in the experimental products. The present study evaluates the application and stability of these formulations in butter, cheese, paneer, biscuit cream, icing cream. The products were evaluated for stability of colour and bixin during storage period. The oil/water soluble propylene glycol formulation was found to be tailor-made for all the dairy products studied though the standardized levels varied between 3.75 and 5.0 mg/kg for butter, 3.75 mg/kg for cheese, 5.0-400 mg/kg for biscuit cream, 12.25 and 25 mg/kg for paneer, 5.0-500 mg/kg for icing cream. Increasing concentration in the range of 30-500 mg/kg yielded products with light cream to orange shades, useful for decorating cakes. The average recovery of bixin from the products immediately after processing was 90-98 %. Significant changes (P ≤ 0.05) were noticed in the colour units and recovery of bixin in all the products during storage of 6 months.


Predicting gene promoter methylation in non-small-cell lung cancer by evaluating sputum and serum.
The use of 5-methylcytosine demethylating agents in conjunction with inhibitors of histone deacetylation may offer a new therapeutic strategy for lung cancer. Monitoring the efficacy of gene demethylating treatment directly within the tumour may be difficult due to tumour location. This study determined the positive and negative predictive values of sputum and serum for detecting gene methylation in primary lung cancer. A panel of eight genes was evaluated by comparing methylation detected in the primary tumour biopsy to serum and sputum obtained from 72 patients with Stage III lung cancer. The prevalence for methylation of the eight genes in sputum (21-43%) approximated to that seen in tumours, but was 0.7-4.3-fold greater than detected in serum. Sputum was superior to serum in classifying the methylation status of genes in the tumour biopsy. The positive predictive value of the top four genes (p16, DAPK, PAX5 beta, and GATA5) was 44-72% with a negative predictive value for these genes > or =70%. The highest specificity was seen for the p16 gene, and this was associated with a odds ratio of six for methylation in the tumour when this gene was methylated in sputum. In contrast, for serum, the individual sensitivity for all genes was 6-27%. Evaluating the combined effect of methylation of at least one of the four most significant genes in sputum increased the positive predictive value to 86%. These studies demonstrate that sputum can be used effectively as a surrogate for tumour tissue to predict the methylation status of advanced lung cancer where biopsy is not feasible.


Probiotics for Antibiotic-Associated Diarrhoea (PAAD): a prospective observational study of antibiotic-associated diarrhoea (including Clostridium difficile-associated diarrhoea) in care homes.
Antibiotic prescribing rates in care homes are higher than in the general population. Antibiotics disrupt the normal gut flora, sometimes causing antibiotic-associated diarrhoea (AAD). Clostridium difficile (Hall and O'Toole 1935) Prévot 1938 is the most commonly identified cause of AAD. Little is known either about the frequency or type of antibiotics prescribed in care homes or about the incidence and aetiology of AAD in this setting. The Probiotics for Antibiotic-Associated Diarrhoea (PAAD) study was designed as a two-stage study. PAAD stage 1 aimed to (1) prospectively describe antibiotic prescribing in care homes; (2) determine the incidence of C. difficile carriage and AAD (including C. difficile-associated diarrhoea); and (3) to consider implementation challenges and establish the basis for a sample size estimation for a randomised controlled trial (RCT) of probiotic administration with antibiotics to prevent AAD in care homes. If justified by PAAD stage 1, the RCT would be implemented in PAAD stage 2. However, as a result of new evidence regarding the clinical effectiveness of probiotics on the incidence of AAD, a decision was taken not to proceed with PAAD stage 2. PAAD stage 1 was a prospective observational cohort study in care homes in South Wales with up to 12 months' follow-up for each resident. Recruited care homes had management and owner's agreement to participate and three or more staff willing to take responsibility for implementing the study. Eleven care homes were recruited, but one withdrew before any residents were recruited. A total of 279 care home residents were recruited to the observational study and 19 withdrew, 16 (84%) because of moving to a non-participating care home. The primary outcomes were the rate of antibiotic prescribing, incidence of AAD, defined as three or more loose stools (type 5-7 on the Bristol Stool Chart) in a 24-hour period, and C. difficile carriage confirmed on stool culture. Stool samples were obtained at study entry from 81% of participating residents. Over half of the samples contained antibiotic-resistant isolates, with Enterobacteriaceae resistant to ciprofloxacin in 47%. Residents were prescribed an average of 2.16 antibiotic prescriptions per year [95% confidence interval (CI) 1.90 to 2.46]. Antibiotics were less likely to be prescribed to residents from dual-registered homes. The incidence of AAD was 0.57 (95% CI 0.41 to 0.81) episodes per year among those residents who were prescribed antibiotics. AAD was more likely in residents who were prescribed co-amoxiclav than other antibiotics and in those residents who routinely used incontinence pads. AAD was less common in residents from residential homes. Care home residents, particularly in nursing homes, are frequently prescribed antibiotics and often experience AAD. Antibiotic resistance, including ciprofloxacin resistance, is common in Enterobacteriaceae isolated from the stool of care home residents. Co-amoxiclav is associated with greater risk of AAD than other commonly prescribed antibiotics. Current Controlled Trials ISRCTN 7954844. This project was funded by the National Institute for Health Research (NIHR) Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 18, No. 63. See the NIHR Journals Library website for further project information.


Toward precision medicine and health: Opportunities and challenges in allergic diseases.
Precision medicine (also called personalized, stratified, or P4 medicine) can be defined as the tailoring of preventive measures and medical treatments to the characteristics of each patient to obtain the best clinical outcome for each person while ideally also enhancing the cost-effectiveness of such interventions for patients and society. Clearly, the best clinical outcome for allergic diseases is not to get them in the first place. To emphasize the importance of disease prevention, a critical component of precision medicine can be referred to as precision health, which is defined herein as the use of all available information pertaining to specific subjects (including family history, individual genetic and other biometric information, and exposures to risk factors for developing or exacerbating disease), as well as features of their environments, to sustain and enhance health and prevent the development of disease. In this article I will provide a personal perspective on how the precision health-precision medicine approach can be applied to the related goals of preventing the development of allergic disorders and providing the most effective diagnosis, disease monitoring, and care for those with these prevalent diseases. I will also mention some of the existing and potential challenges to achieving these ambitious goals.


The dimanganese(II) site of Bacillus subtilis class Ib ribonucleotide reductase.
Class Ib ribonucleotide reductases (RNRs) use a dimanganese-tyrosyl radical cofactor, Mn(III)(2)-Y(•), in their homodimeric NrdF (β2) subunit to initiate reduction of ribonucleotides to deoxyribonucleotides. The structure of the Mn(II)(2) form of NrdF is an important component in understanding O(2)-mediated formation of the active metallocofactor, a subject of much interest because a unique flavodoxin, NrdI, is required for cofactor assembly. Biochemical studies and sequence alignments suggest that NrdF and NrdI proteins diverge into three phylogenetically distinct groups. The only crystal structure to date of a NrdF with a fully ordered and occupied dimanganese site is that of Escherichia coli Mn(II)(2)-NrdF, prototypical of the enzymes from actinobacteria and proteobacteria. Here we report the 1.9 Å resolution crystal structure of Bacillus subtilis Mn(II)(2)-NrdF, representative of the enzymes from a second group, from Bacillus and Staphylococcus. The structures of the metal clusters in the β2 dimer are distinct from those observed in E. coli Mn(II)(2)-NrdF. These differences illustrate the key role that solvent molecules and protein residues in the second coordination sphere of the Mn(II)(2) cluster play in determining conformations of carboxylate residues at the metal sites and demonstrate that diverse coordination geometries are capable of serving as starting points for Mn(III)(2)-Y(•) cofactor assembly in class Ib RNRs.


The biogenic approach to cognition.
After half a century of cognitive revolution we remain far from agreement about what cognition is and what cognition does. It was once thought that these questions could wait until the data were in. Today there is a mountain of data, but no way of making sense of it. The time for tackling the fundamental issues has arrived. The biogenic approach to cognition is introduced not as a solution but as a means of approaching the issues. The traditional, and still predominant, methodological stance in cognitive inquiry is what I call the anthropogenic approach: assume human cognition as the paradigm and work 'down' to a more general explanatory concept. The biogenic approach, on the other hand, starts with the facts of biology as the basis for theorizing and works 'up' to the human case by asking psychological questions as if they were biological questions. Biogenic explanations of cognition are currently clustered around two main frameworks for understanding biology: self-organizing complex systems and autopoiesis. The paper describes the frameworks and infers from them ten empirical principles--the biogenic 'family traits'--that constitute constraints on biogenic theorizing. Because the anthropogenic approach to cognition is not constrained empirically to the same degree, I argue that the biogenic approach is superior for approaching a general theory of cognition as a natural phenomenon.


Alcohol use after forced abstinence in basic training among United States Navy and Air Force trainees.
The present study examined drinking rates of U.S. Navy and Air Force trainees during the month before attending basic training and after the ban on drinking alcohol was lifted during advanced training. Surveys were obtained at five training bases (one U.S. Navy, four U.S. Air Force) from 6,298 persons for a response rate of 65%. Analyses were based on 4,962 young adults ages 18-25 (82% male) who gave valid responses about pre-basic drinking. Findings showed a rate of 43.1% heavy episodic drinking during the month before basic training, 15.8% by infrequent heavy episodic drinkers (five or more drinks per occasion at least once, four or more for women), and 27.3% by frequent heavy episodic drinkers (five or more drinks per occasion, four or more for women, at least once a week). Pre-basic frequent heavy episodic drinkers averaged 7.9 drinks per occasion and nearly 15 heavy episodic drinking days during the month. In contrast, heavy episodic drinking following basic training was substantially lower: 12.0% for infrequent heavy episodic drinkers and 9.0% for frequent heavy episodic drinkers. Comparisons with the U.S. Department of Defense Health Related Behaviors Survey suggest that post-basic frequent heavy episodic drinking rates are likely to increase over time but not to pre-basic levels. Pre-basic infrequent and frequent heavy episodic drinkers were more likely than nondrinkers or non-heavy episodic drinkers to initiate or re-initiate frequent heavy episodic drinking. Frequent heavy episodic drinking after basic tended to occur 1-6 weeks after the ban on alcohol use was lifted. Selection and socialization help explain heavy episodic drinking of U.S Navy and U.S. Air Force trainees.


A mutation in the Na-K-2Cl cotransporter-1 leads to changes in cellular metabolism.
The Na-K-Cl cotransporter-1 (NKCC1), by mediating the electroneutral transport of Na+ , K+ , and Cl- plays an important role in cell volume regulation, epithelial transport, and the control of neuronal excitability. Recently, we reported the first known human mutation in SLC12A2, the gene encoding NKCC1. The 17-year old patient suffers from multiorgan failure. Laboratory tests conducted on muscle and liver biopsies of the patient showed abnormal increase in mitochondrial DNA copy number and increased glycogen levels, indicating the possibility that the transporter may play a role in energy metabolism. Here, we show that fibroblasts isolated from the patient demonstrate a significant increase in mitochondrial respiration, compared to fibroblasts isolated from healthy individuals. Similarly, Madin Darby canine kidney (MDCK) cells transfected with enhanced green fluorescent protein (EGFP)-tagged mutant NKCC1 DNA demonstrated increased mitochondrial respiration when compared to MDCK cells expressing EGFP-tagged wild-type (WT) cotransporter. Direct inhibition of the cotransporter through addition of bumetanide did not change the rate of basal respiration, but led to increased maximal mitochondrial respiration. Fibroblasts extracted from NKCC1WT/DFX and NKCC1DFX/DFX mice also demonstrated a significant elevation in mitochondrial respiration, compared to fibroblasts isolated from their WT littermates. Expression of the mutant protein was associated with an increase in hydrogen peroxide and peroxidase activity and a decrease in messenger RNA transcript levels for protein involved in the unfolded protein response. These data reveal that cells expressing the mutant cotransporter demonstrate increased mitochondrial respiration and behave like they are experiencing a state of starvation.


HtrA1-dependent proteolysis of TGF-beta controls both neuronal maturation and developmental survival.
Transforming growth factor-beta (TGF-beta) signalling controls a number of cerebral functions and dysfunctions including synaptogenesis, amyloid-beta accumulation, apoptosis and excitotoxicity. Using cultured cortical neurons prepared from either wild type or transgenic mice overexpressing a TGF-beta-responsive luciferase reporter gene (SBE-Luc), we demonstrated a progressive loss of TGF-beta signalling during neuronal maturation and survival. Moreover, we showed that neurons exhibit increasing amounts of the serine protease HtrA1 (high temperature responsive antigen 1) and corresponding cleavage products during both in vitro neuronal maturation and brain development. In parallel of its ability to promote degradation of TGF-beta1, we demonstrated that blockage of the proteolytic activity of HtrA1 leads to a restoration of TGF-beta signalling, subsequent overexpression of the serpin type -1 plasminogen activator inhibitor (PAI-1) and neuronal death. Altogether, we propose that the balance between HtrA1 and TGF-beta could be one of the critical events controlling both neuronal maturation and developmental survival.


Outcomes in octogenarians undergoing high-risk cancer operation: a national study.
Current information about outcomes in octogenarians undergoing cancer operations is limited largely to case series from selected centers. Population-based data can provide more realistic estimates of the risks and benefits of operations in this group. We performed a retrospective cohort study of patients undergoing major resections for lung, esophageal, and pancreas cancer. Using the Nationwide Inpatient Sample (1994 to 2003), we examined operative mortality and discharge disposition in octogenarians (aged 80+ years), relative to younger patients (aged 65 to 69 years) (n = 272,662). We then used the Surveillance and End Results-Medicare-linked database (1992 to 2001) to measure late survival in the elderly (n = 14,088). Operative mortality among octogenarians was substantially higher than that of younger patients (aged 65 to 69 years) for all three cancers (esophagectomy, 19.9% versus 8.8%, p < 0.0001; pancreatectomy, 15.5% versus 6.7%, p < 0.0001; lung resection, 6.9% versus 3.7%, p < 0.0001). A large proportion of octogenarians were transferred to extended care facilities after operation, ranging from 24% after lung resection to 44% after esophagectomy. Five-year survival in octogenarians was low for all three cancers: 11% after pancreatectomy, 18% after esophagectomy and 31% after lung cancer resection. Survival among octogenarians with two or more comorbidities was worse than those with fewer comorbid diagnoses--10% versus 14% for pancreatectomy, 15% versus 23% for esophagectomy, and 27% versus 37% for lung resection. Population-based outcomes after high-risk cancer operation in octogenarians are considerably worse than typically reported in case series and published survival statistics. Such information might better inform clinical decision making in this high-risk group.


Endogenous glucagon-like peptide 1 controls endocrine pancreatic secretion and antro-pyloro-duodenal motility in humans.
Exogenous use of the intestinal hormone glucagon-like peptide 1 (GLP-1) lowers glycaemia by stimulation of insulin, inhibition of glucagon, and delay of gastric emptying. To assess the effects of endogenous GLP-1 on endocrine pancreatic secretion and antro-pyloro-duodenal motility by utilising the GLP-1 receptor antagonist exendin(9-39)amide (ex(9-39)NH2). Nine healthy volunteers underwent four experiments each. In two experiments with and without intravenous infusion of ex(9-39)NH2 300 pmol/kg/min, a fasting period was followed by intraduodenal glucose perfusion at 1 and 2.5 kcal/min, with the higher dose stimulating GLP-1 release. Antro-pyloro-duodenal motility was measured by perfusion manometry. To calculate the incretin effect (that is, the proportion of plasma insulin stimulated by intestinal hormones) the glycaemia observed during the luminal glucose experiments was mimicked using intravenous glucose in two further experiments. Ex(9-39)NH2 significantly increased glycaemia during fasting and duodenal glucose. It diminished plasma insulin during duodenal glucose and significantly reduced the incretin effect by approximately 50%. Ex(9-39)NH2 raised plasma glucagon during fasting and abolished the decrease in glucagon at the high duodenal glucose load. Ex(9-39)NH2 markedly stimulated antroduodenal contractility. At low duodenal glucose it reduced the stimulation of tonic and phasic pyloric motility. At the high duodenal glucose load it abolished pyloric stimulation. Endogenous GLP-1 stimulates postprandial insulin release. The pancreatic alpha cell is under the tonic inhibitory control of GLP-1 thereby suppressing postprandial glucagon. GLP-1 tonically inhibits antroduodenal motility and mediates the postprandial inhibition of antral and stimulation of pyloric motility. We therefore suggest GLP-1 as a true incretin hormone and enterogastrone in humans.


Modeling present and future climate risk of dengue outbreak, a case study in New Caledonia.
Dengue dynamics result from the complex interactions between the virus, the host and the vector, all being under the influence of the environment. Several studies explored the link between weather and dengue dynamics and some investigated the impact of climate change on these dynamics. Most attempted to predict incidence rate at a country scale or assess the environmental suitability at a global or regional scale. Here, we propose a new approach which consists in modeling the risk of dengue outbreak at a local scale according to climate conditions and study the evolution of this risk taking climate change into account. We apply this approach in New Caledonia, where high quality data are available. We used a statistical estimation of the effective reproduction number (Rt) based on case counts to create a categorical target variable : epidemic week/non-epidemic week. A machine learning classifier has been trained using relevant climate indicators in order to estimate the probability for a week to be epidemic under current climate data and this probability was then estimated under climate change scenarios. Weekly probability of dengue outbreak was best predicted with the number of days when maximal temperature exceeded 30.8°C and the mean of daily precipitation over 80 and 60 days prior to the predicted week respectively. According to scenario RCP8.5, climate will allow dengue outbreak every year in New Caledonia if the epidemiological and entomological contexts remain the same. We identified locally relevant climatic factor driving dengue outbreaks in New Caledonia and assessed the inter-annual and seasonal risk of dengue outbreak under different climate change scenarios up to the year 2100. We introduced a new modeling approach to estimate the risk of dengue outbreak depending on climate conditions. This approach is easily reproducible in other countries provided that reliable epidemiological and climate data are available.


Pancreatic cancer incidence in relation to female reproductive factors: Iowa Women's Health Study.
Motivated by inconsistent literature, we evaluated the association between incident pancreatic cancer and reproductive characteristics. The Iowa Women's Health Study is a large prospective population-based cohort followed from 1986 to 2003. Reproductive information was self-reported. The study population comprised 37,459 women aged 55-69 years at baseline. Over 18 years, 228 incident pancreatic cancers were identified. In a multivariate-adjusted model there were no associations between incident pancreatic cancer and age at first birth, number of births, age at menarche, or use of hormones. There was a statistically significant inverse association between age at menopause and pancreatic cancer incidence. Compared to menopause less than 45 years, the hazard ratio of pancreatic cancer was 0.61 (95% CI, 0.40-0.94) for menopause at 45-49 years, 0.75 (95% CI, 0.51-1.09) for 50-54 years, and 0.35 (95% CI, 0.18-0.68) for menopause at 55 years or more (P trend=0.005). This association held after restricting the cohort to never smokers. The associations between pancreatic cancer and ages at natural and surgical menopause followed similar patterns. In a parallel fashion, risk of pancreatic cancer was decreased for women with intact ovaries compared to those who had oophorectomy: hazard ratio was 0.70 (95% CI, 0.50-0.99). Our results indicate that older age at menopause is associated with reduced pancreatic cancer risk, but further research is warranted.


Photoinactivation of uncultured, indigenous enterococci.
Enterococci are used to monitor recreational water quality worldwide, so understanding their fate and transport in the environment is essential to the protection of human health. As such, researchers have documented enterococci inactivation under various exposure conditions and in diverse water matrices. However, the majority of studies have been performed using lab-cultured bacteria, which are distinct from indigenous, uncultured bacteria found in the environment. Here we investigate the photoinactivation of indigenous, uncultured enterococci from a range of sources, including wastewater treatment plants (WWTPs), marine beaches, urban streams, and a wastewater-influenced pond. We concentrated indigenous enterococci from their sources using filtration and centrifugation, placed them in a clear buffer solution, and then exposed them to simulated sunlight to measure their photoinactivation rates. First order decay rate constants (k) of indigenous, uncultured enterococci spanned an order of magnitude, from 0.3 to 2.3 m2 kJUVB-1. k values of indigenous enterococci from WWTPs tended to be larger than those from surface waters. The k value of lab-cultured Enterococcus faecalis was larger than those of indigenous, uncultured enterococci from most sources. Negative associations between the fraction of pigmented enterococci and sunlight susceptibility were observed. This work suggests that caution should be taken when extending results on bacterial photoinactivation obtained using lab-cultured bacteria to environmental bacteria, and that enterococci pigmentation may be a useful metric for estimating photoinactivation rate constants.


A map of the protein space--an automatic hierarchical classification of all protein sequences.
We investigate the space of all protein sequences. We combine the standard measures of similarity (SW, FASTA, BLAST), to associate with each sequence an exhaustive list of neighboring sequences. These lists induce a (weighted directed) graph whose vertices are the sequences. The weight of an edge connecting two sequences represents their degree of similarity. This graph encodes much of the fundamental properties of the sequence space. We look for clusters of related proteins in this graph. These clusters correspond to strongly connected sets of vertices. Two main ideas underlie our work: i) Interesting homologies among proteins can be deduced by transitivity. ii) Transitivity should be applied restrictively in order to prevent unrelated proteins from clustering together. Our analysis starts from a very conservative classification, based on very significant similarities, that has many classes. Subsequently, classes are merged to include less significant similarities. Merging is performed via a novel two phase algorithm. First, the algorithm identifies groups of possibly related clusters (based on transitivity and strong connectivity) using local considerations, and merges them. Then, a global test is applied to identify nuclei of strong relationships within these groups of clusters, and the classification is refined accordingly. This process takes place at varying thresholds of statistical significance, where at each step the algorithm is applied on the classes of the previous classification, to obtain the next one, at the more permissive threshold. Consequently, a hierarchical organization of all proteins is obtained. The resulting classification splits the space of all protein sequences into well defined groups of proteins. The results show that the automatically induced sets of proteins are closely correlated with natural biological families and super families. The hierarchical organization reveals finer sub-families that make up known families of proteins as well as many interesting relations between protein families. The hierarchical organization proposed may be considered as the first map of the space of all protein sequences. An interactive web site including the results of our analysis has been constructed, and is now accessible through http:/(/)www.protomap.cs.huji.ac.il


The predictability of evolution: glimpses into a post-Darwinian world.
The very success of the Darwinian explanation, in not only demonstrating evolution from multiple lines of evidence but also in providing some plausible explanations, paradoxically seems to have served to have stifled explorations into other areas of investigation. The fact of evolution is now almost universally yoked to the assumption that its outcomes are random, trends are little more than drunkard's walks, and most evolutionary products are masterpieces of improvisation and far from perfect. But is this correct? Let us consider some alternatives. Is there evidence that evolution could in anyway be predictable? Can we identify alternative forms of biological organizations and if so how viable are they? Why are some molecules so extraordinarily versatile, while others can be spoken of as "molecules of choice"? How fortuitous are the major transitions in the history of life? What implications might this have for the Tree of Life? To what extent is evolutionary diversification constrained or facilitated by prior states? Are evolutionary outcomes merely sufficient or alternatively are they highly efficient, even superb? Here I argue that in sharp contradistinction to an orthodox Darwinian view, not only is evolution much more predictable than generally assumed but also investigation of its organizational substrates, including those of sensory systems, which indicates that it is possible to identify a predictability to the process and outcomes of evolution. If correct, the implications may be of some significance, not least in separating the unexceptional Darwinian mechanisms from underlying organizational principles, which may indicate evolutionary inevitabilities.


Fetal brain tissue annotation and segmentation challenge results.
In-utero fetal MRI is emerging as an important tool in the diagnosis and analysis of the developing human brain. Automatic segmentation of the developing fetal brain is a vital step in the quantitative analysis of prenatal neurodevelopment both in the research and clinical context. However, manual segmentation of cerebral structures is time-consuming and prone to error and inter-observer variability. Therefore, we organized the Fetal Tissue Annotation (FeTA) Challenge in 2021 in order to encourage the development of automatic segmentation algorithms on an international level. The challenge utilized FeTA Dataset, an open dataset of fetal brain MRI reconstructions segmented into seven different tissues (external cerebrospinal fluid, gray matter, white matter, ventricles, cerebellum, brainstem, deep gray matter). 20 international teams participated in this challenge, submitting a total of 21 algorithms for evaluation. In this paper, we provide a detailed analysis of the results from both a technical and clinical perspective. All participants relied on deep learning methods, mainly U-Nets, with some variability present in the network architecture, optimization, and image pre- and post-processing. The majority of teams used existing medical imaging deep learning frameworks. The main differences between the submissions were the fine tuning done during training, and the specific pre- and post-processing steps performed. The challenge results showed that almost all submissions performed similarly. Four of the top five teams used ensemble learning methods. However, one team's algorithm performed significantly superior to the other submissions, and consisted of an asymmetrical U-Net network architecture. This paper provides a first of its kind benchmark for future automatic multi-tissue segmentation algorithms for the developing human brain in utero.


Recurrent Mutations of Chromatin-Remodeling Genes and Kinase Receptors in Pheochromocytomas and Paragangliomas.
Pheochromocytomas and paragangliomas (PPGL) are genetically heterogeneous tumors of neural crest origin, but the molecular basis of most PPGLs is unknown. We performed exome or transcriptome sequencing of 43 samples from 41 patients. A validation set of 136 PPGLs was used for amplicon-specific resequencing. In addition, a subset of these tumors was subjected to microarray-based transcription, protein expression, and histone methylation analysis by Western blotting or immunohistochemistry. In vitro analysis of mutants was performed in cell lines. We detected mutations in chromatin-remodeling genes, including histone-methyltransferases, histone-demethylases, and histones in 11 samples from 8 patients (20%). In particular, we characterized a new cancer syndrome involving PPGLs and giant cell tumors of bone (GCT) caused by a postzygotic G34W mutation of the histone 3.3 gene, H3F3A Furthermore, mutations in kinase genes were detected in samples from 15 patients (37%). Among those, a novel germline kinase domain mutation of MERTK detected in a patient with PPGL and medullary thyroid carcinoma was found to activate signaling downstream of this receptor. Recurrent germline and somatic mutations were also detected in MET, including a familial case and sporadic PPGLs. Importantly, in each of these three genes, mutations were also detected in the validation group. In addition, a somatic oncogenic hotspot FGFR1 mutation was found in a sporadic tumor. This study implicates chromatin-remodeling and kinase variants as frequent genetic events in PPGLs, many of which have no other known germline driver mutation. MERTK, MET, and H3F3A emerge as novel PPGL susceptibility genes. Clin Cancer Res; 22(9); 2301-10. ©2015 AACR.


Behavioural weight management programmes for adults assessed by trials conducted in everyday contexts: systematic review and meta-analysis.
This systematic review and meta-analysis of effectiveness trials comparing multicomponent behavioural weight management programmes with controls in overweight and obese adults set out to determine the effectiveness of these interventions implemented in routine practice. To be included, interventions must have been multicomponent, delivered by the therapists who would deliver the intervention in routine practice and in that same context, and must be widely available or feasible to implement with little additional infrastructure or staffing. Searches of electronic databases were conducted, and augmented by screening reference lists and contacting experts (November 2012). Data were extracted by two reviewers, with mean difference between intervention and control for 12-month change in weight, blood pressure, lipids and glucose calculated using baseline observation carried forward. Data were also extracted on adverse events, quality of life and mood measures. Although there were many published efficacy trials, only eight effectiveness trials met the inclusion criteria. Pooled results from five study arms providing access to commercial weight management programmes detected significant weight loss at 12 months (mean difference -2.22 kg, 95% confidence interval [CI] -2.90 to -1.54). Results from two arms of a study testing a commercial programme providing meal replacements also detected significant weight loss (mean difference -6.83 kg, 95% CI -8.39 to -5.26). In contrast, pooled results from five interventions delivered by primary care teams showed no evidence of an effect on weight (mean difference -0.45 kg, 95% CI -1.34 to 0.43). One study testing an interactive web-based intervention detected a significant effect in favour of the intervention at 12 months, but the study was judged to be at high risk of bias and the effect did not persist at 18 months. Few studies reported other outcomes, limiting comparisons between interventions. Few trials have examined the effectiveness of behavioural weight loss programmes delivered in everyday contexts. These trials suggest that commercial interventions delivered in the community are effective for achieving weight loss. There is no evidence that interventions delivered within primary care settings by generalist primary care teams trained in weight management achieve meaningful weight loss.


Is bovine dentine an appropriate substitute for human dentine in erosion/abrasion tests?
The study aimed to compare the dentine wear of primary and permanent human and bovine teeth because of erosion/abrasion and evaluate if bovine dentine is an appropriate substitute for human dentine in further erosion/abrasions tests. Dentine samples from deciduous molars and human third molars as well as from calves' and cattle's lower incisors were prepared and baseline surface profiles were recorded. Each day all samples were demineralized in 1% citric acid, tooth brushed with 100 brushing strokes with toothpaste slurry and stored in artificial saliva for the rest of the day. This cycle was run for 20 days. Afterwards, new surface profiles were recorded and dentine wear was calculated by a customized computer program. Dentine wear because of erosion/abrasion was not statistically, significantly different for human third molars and cattle's lower incisors (P = 0.7002). The dentine wear because of erosion/abrasion of deciduous molars and calves' lower incisors was significantly different (P < 0.0000). No statistically significant difference in the dentine wear of human third molars and cattle's lower incisors was observed, so that the use of cattle's lower incisors as substitute for adult human teeth for further investigations in erosion/abrasion studies could be accepted.


Near-isometric flattening of brain surfaces.
Flattened representations of brain surfaces are often used to visualize and analyze spatial patterns of structural organization and functional activity. Here, we present a set of rigorous criteria and accompanying test cases with which to evaluate flattening algorithms that attempt to preserve shortest-path distances on the original surface. We also introduce a novel flattening algorithm that is the first to satisfy all of these criteria and demonstrate its ability to produce accurate flat maps of human and macaque visual cortex. Using this algorithm, we have recently obtained results showing a remarkable, unexpected degree of consistency in the shape and topographic structure of visual cortical areas within humans and macaques, as well as between these two species.


Comparison of the proteins in salivary glands, saliva and haemolymph of Rhipicephalus appendiculatus female ticks during feeding.
To compare the proteins in salivary glands, saliva and haemolymph of Rhipicephalus appendiculatus female ticks, antisera were prepared from guinea-pigs immunized with soluble denatured salivary gland extracts (SGE). The extracts were derived from R. appendiculatus female ticks that were either unfed (day 0) or partly fed (day 6). The sera were used in immunoblotting, following SDS-polyacrylamide gel electrophoresis, to examine the antigen profiles during the course of tick feeding on guinea-pigs. Day 0 and day 6 SGE antisera appeared to detect common proteins in the different tick samples. For example, haemolymph apparently shared some of the small protein bands (31.5-34 kDa) detected in SGEs. These small proteins appeared in both samples at the same stage of feeding, suggesting that haemolymph and salivary glands not only have common antigens but may also share some functions. Furthermore, a number of protein bands were detected in haemolymph before they were apparent in the salivary glands or saliva. Thus some antigens detected in the salivary glands and saliva may be derived from the haemolymph. The results indicate that the host may be exposed to tick saliva antigens that are also present in the haemolymph. We discuss the significance of these observations with regard to the induction of host immunity to ticks and the development of tick vaccines.


Evidence for the presence of oxidatively modified low density lipoprotein in atherosclerotic lesions of rabbit and man.
Three lines of evidence are presented that low density lipoproteins gently extracted from human and rabbit atherosclerotic lesions (lesion LDL) greatly resembles LDL that has been oxidatively modified in vitro. First, lesion LDL showed many of the physical and chemical properties of oxidized LDL, properties that differ from those of plasma LDL: higher electrophoretic mobility, a higher density, higher free cholesterol content, and a higher proportion of sphingomyelin and lysophosphatidylcholine in the phospholipid fraction. A number of lower molecular weight fragments of apo B were found in lesion LDL, similar to in vitro oxidized LDL. Second, both the intact apo B and some of the apo B fragments of lesion LDL reacted in Western blots with antisera that recognize malondialdehyde-conjugated lysine and 4-hydroxynonenal lysine adducts, both of which are found in oxidized LDL; plasma LDL and LDL from normal human intima showed no such reactivity. Third, lesion LDL shared biological properties with oxidized LDL: compared with plasma LDL, lesion LDL produced much greater stimulation of cholesterol esterification and was degraded more rapidly by macrophages. Degradation of radiolabeled lesion LDL was competitively inhibited by unlabeled lesion LDL, by LDL oxidized with copper, by polyinosinic acid and by malondialdehyde-LDL, but not by native LDL, indicating uptake by the scavenger receptor(s). Finally, lesion LDL (but not normal intimal LDL or plasma LDL) was chemotactic for monocytes, as is oxidized LDL. These studies provide strong evidence that atherosclerotic lesions, both in man and in rabbit, contain oxidatively modified LDL.


[Epidemiological situation of visceral leishmaniasis on the Island of São Luis, State of Maranhão].
The results from a field study on autochthonous visceral leishmaniasis on the island of São Luís are presented. This study started in 2004 and finished in 2006 and had the aim of ascertaining the determinant epidemiological and clinical characteristics of this endemic disease. Two hundred ninety nine autochthonous cases were analyzed, of which 83.6% were children younger than 9 years old and 54.1% were male. The disease occurred in all months of the year, with a peak in June. The coefficient of incidence decreased from 46.1 to 35.2 cases per 100,000 inhabitants over the years studied. The diagnosis was confirmed by laboratory tests in 93.3% of the cases. The treatment of choice was based on N-methylglucamine, with a cure rate of 96.1%. The mean lethality rate was 3.7%. Because of the absence of systematic control actions, the authors propose the creation of a specific program to be developed by the municipalities under the coordination of the State Department of Health.


Using the epigenetic field defect to detect prostate cancer in biopsy negative patients.
We determined whether a novel combination of field defect DNA methylation markers could predict the presence of prostate cancer using histologically normal transrectal ultrasound guided biopsy cores. Methylation was assessed using quantitative Pyrosequencing® in a training set consisting of 65 nontumor and tumor associated prostate tissues from University of Wisconsin. A multiplex model was generated using multivariate logistic regression and externally validated in blinded fashion in a set of 47 nontumor and tumor associated biopsy specimens from University of Washington. We observed robust methylation differences in all genes at all CpGs assayed (p <0.0001). Regression models incorporating individual genes (EVX1, CAV1 and FGF1) and a gene combination (EVX1 and FGF1) discriminated nontumor from tumor associated tissues in the original training set (AUC 0.796-0.898, p <0.001). On external validation uniplex models incorporating EVX1, CAV1 or FGF1 discriminated tumor from nontumor associated biopsy negative specimens (AUC 0.702, 0.696 and 0.658, respectively, p <0.05). A multiplex model (EVX1 and FGF1) identified patients with prostate cancer (AUC 0.774, p = 0.001) and had a negative predictive value of 0.909. Comparison between 2 separate cores in patients in this validation set revealed similar methylation defects, indicating detection of a widespread field defect. A widespread epigenetic field defect can be used to detect prostate cancer in patients with histologically negative biopsies. To our knowledge this assay is unique, in that it detects alterations in nontumor cells. With further validation this marker combination (EVX1 and FGF1) has the potential to decrease the need for repeat prostate biopsies, a procedure associated with cost and complications.


Biomechanical Deficit Profiles Associated with ACL Injury Risk in Female Athletes.
This study aimed to quantify the prevalence of biomechanical deficit patterns associated with anterior cruciate ligament injury risk and their interconnections in a large cohort of female athletes during an unanticipated cutting task. High school female athletes (n = 721) performed an unanticipated cutting task in the biomechanics laboratory. Trunk and lower extremity three-dimensional kinetics and kinematics were measured and entered into a latent profile analysis model. Approximately 40% of female athletes demonstrated no biomechanical deficits and were categorized into the low risk group. The second most prevalent profile (24%) demonstrated a combination of high quadriceps and leg dominance deficits and was labeled as quadriceps-leg. The third most prevalent profile (22%) demonstrated a combination of trunk and leg dominance deficits and to lesser extent ligament dominance deficits and was labeled as trunk-leg-ligament. Finally, the fourth profile (14%) demonstrated very high ligament dominance deficits only and it was labeled as ligament dominance profile. This is the first study to identify the most common biomechanical profiles associated with anterior cruciate ligament injury during a cutting task in a large cohort of female athletes. Approximately 60% of female athletes belong to one of the high-risk profiles. With the exception of the ligament dominance profile, the current analysis indicates that risk profiles consist of a combination of biomechanical deficits. The findings provide important insight into the prevalence of biomechanical deficits and future directions for the development of injury prevention programs. The findings can be used to guide the development of quick and easy tests that accurately categorize athletes into one of the profiles and subsequently prescribe tailored injury prevention programs that will be more effective and efficient than the current generic ones.