The relationship between palmitoyl-coenzyme A synthetase activity and esterification of sn-glycerol 3-phosphate by the microsomal fraction of guinea-pig intestinal mucosa.
1. With microsomal fractions of guinea-pig intestinal mucosa the mean specific activity of palmitoyl-CoA synthetase was approx. 1.3-fold the esterification of sn-glycerol 3-phosphate with palmitoyl-CoA generated by the endogenous synthetase. The latter activity was approx. 2.5- and 5-fold that when palmitoyl-CoA was generated from palmitoylcarnitine or when it was added directly to the assay system. 2. There were significant correlations (P<0.001) between the specific activities of palmitoyl-CoA synthetase and glycerolipid synthesis from either palmitate or palmitoylcarnitine. 3. The mean molar composition of glycerolipid synthesized from palmitate or palmitoylcarnitine was approx. 18% lysophosphatidate, 75% phosphatidate and 7% neutral lipid. 4. Glycerolipid synthesis from palmitate was inhibited by 80-90% after preincubation of microsomal fractions at 37 degrees C for 40min and was caused by inactivation of palmitoyl-CoA synthetase. 5. Addition of 100-400mm-KCl inhibited palmitoyl-CoA synthetase activity and glycerolipid synthesis from palmitate but stimulated glycerol phosphate acyltransferase activity. 6. Diversion of palmitoyl-CoA synthesized by the endogenous synthetase to palmitoylcarnitine resulted in an almost stoicheiometric decrease in glycerolipid synthesis. 7. Addition of rac-1-monopalmitin promoted utilization of palmitoyl-CoA by the monoglyceride pathway but did not inhibit phosphatidate biosynthesis. 8. With rate-limiting concentrations of CoA and Mg(2+) the relative decreases in velocity for palmitoyl-CoA synthetase and glycerolipid synthesis from palmitate were almost identical. However, low concentrations of palmitate and ATP produced greater decreases in synthetase activity than in glycerolipid synthesis. 9. There appears to be a fine balance between the activities of palmitoyl-CoA synthetase and glycerol phosphate acyltransferase, with neither activity being in excess with respect to phosphatidate synthesis.


The pattern of astigmatism in a Canadian preschool population.
To measure magnitude, type, and central tendency of astigmatism found in a county-wide population of Canadian preschool children (mean age, 48.1 months). Noncycloplegic autorefractive measures were taken in 1179 children attending a preschool health fair operated by their county board of health. Spherocylinder measures were transformed into three independent components. The equivalent sphere showed considerable variation between retinoscopy and autorefraction that was attributed to the variable overaccommodation induced by the autorefractor. Astigmatic components were not affected. Small discrepancies between the two techniques were similar to those in adults and were not of sufficient magnitude to affect validity. With-the-rule (WTR) astigmatism of at least 0.25 D was the most frequent form (45%) followed by against-the-rule (ATR; 40%) and oblique (15%). The 95th percentile for cylinder magnitude was found at 1.25 D. Astigmatisms beyond this value were predominately WTR. The mean (negative) cylinder magnitude was 0.08 Dx 015 degrees. When spherocylinder values are transformed into a mathematical continuum rather than WTR and ATR classifications, the true central tendency of the population is better defined and is close to zero. Astigmatisms of more than 1.25 D in the preschool child exceed the 95th percentile in this population and were more frequently WTR.


Near-chromosome level genome assembly of the fruit pest Drosophila suzukii using long-read sequencing.
Over the past decade, the spotted wing Drosophila, Drosophila suzukii, has invaded Europe and America and has become a major agricultural pest in these areas, thereby prompting intense research activities to better understand its biology. Two draft genome assemblies already exist for this species but contain pervasive assembly errors and are highly fragmented, which limits their values. Our purpose here was to improve the assembly of the D. suzukii genome and to annotate it in a way that facilitates comparisons with D. melanogaster. For this, we generated PacBio long-read sequencing data and assembled a novel, high-quality D. suzukii genome assembly. It is one of the largest Drosophila genomes, notably because of the expansion of its repeatome. We found that despite 16 rounds of full-sib crossings the D. suzukii strain that we sequenced has maintained high levels of polymorphism in some regions of its genome. As a consequence, the quality of the assembly of these regions was reduced. We explored possible origins of this high residual diversity, including the presence of structural variants and a possible heterogeneous admixture pattern of North American and Asian ancestry. Overall, our assembly and annotation constitute a high-quality genomic resource that can be used for both high-throughput sequencing approaches, as well as manipulative genetic technologies to study D. suzukii.


A family tradition of Scottish medicine.
In the late nineteenth and early twentieth centuries, Edinburgh was among the most popular of all destinations for South Africans seeking a medical education. Scotland was thus the fountainhead of much of the knowledge and skill that subsequently led to the development of a high standard of medical education in South Africa. This article describes how four members of a South African family received their medical education at the University and Royal Colleges of Physicians and Surgeons of Edinburgh and a fifth family member obtained the Membership of the Royal College of Physicians and was later elected to Fellowship. This is the story of a relationship spanning three generations between a family in which medicine had almost become a hereditary trait and the best of Scottish medical education.


Fatty acid, isoprenoid quinone and polar lipid composition in the classification of Curtobacterium and related taxa.
Strains representing the taxa Curtobacterium, Brevibacterium saperdae, B. testaceum, Corynebacterium betae, Cor. nebraskense and Cor. oortii were degraded by acid methanolysis and the non-hydroxylated fatty acid esters released were examined by thin-layer and gas-liquid chromatography. The major fatty acids in all strains were 12-methyltetradecanoic (anteiso C15) and 14-methylhexadecanoic (anteiso C17) acids which occurred together with other anteiso, iso and straight-chain acids. Polar lipids of the test strains were examined by two-dimensional thin-layer chromatography. All organisms possessed very characteristic polar lipid patterns consisting of diphosphatidylglycerol, phosphatidylglycerol and a number of uncharacterized glycolipids. The menaquinone components of the test strains facilitated their division into two groups containing, respectively, nine isoprene units (viz. Curtobacterium citreum, Curt. luteum, Corynebacterium betae, Cor. flaccumfaciens, Cor. oortii, Cor. poinsettiae and Cor. nebraskense) and eleven and twelve isoprene units (viz. Brevibacterium saperdae, B. testaceum). The results of the present study indicate that the genus Curtobacterium as presently recognized is a heterogeneous taxon containing two distinct centres of variation.


Medical and graduate students' attitudes toward personal genomics.
Medical schools are being approached by direct-to-consumer genotyping companies about genotyping faculty or trainees as a method to "teach" them about the potential implications of genotyping. In thinking about the future incorporation of genotyping into a graduate level genetics course, the purpose of this study was 2-fold: first, to assess knowledge, attitudes, and beliefs of students toward personal genomics as it related to themselves as both as customers and future physicians and as it related to consumers at large, and second, to determine the impact of the course (as taught without genotyping) on knowledge, attitudes, and beliefs. We surveyed first-year medical students and graduate students before and after a core genetics course. After the course, students were less likely to believe that genotyping information would be useful to physicians, patients, or consumers; genotyping would provide information to improve their own personal health; or personal genomic testing services are diagnostic of medical conditions. They were more likely to answer knowledge questions accurately after the course but still had difficulty with clinical interpretation. Despite these changes, a slight majority of students were, and remained, interested in undergoing genotyping themselves. Of note, the number who believed genotyping "would help them understand genetic concepts better than someone else's data" decreased. General curiosity was the most commonly chosen reason for interest in undergoing genotyping, and approximately 50% of respondents expressed concern about confidentiality of results. In conclusion, even without the genotyping process, an educational program about genotyping increased knowledge, particularly about the clinical limitations of genotyping, but student interest in genotyping did not significantly change. Institutions thinking about offering genotyping to their students as part of a learning experience should consider the pros and cons of doing so.


Desmopressin add-on therapy for refractory nocturia in men receiving α-blockers for lower urinary tract symptoms.
Alpha-blockers improve lower urinary tract symptoms associated with benign prostatic obstruction. Nocturia, a storage symptom, is a common complaint in men. However, it does not fully respond to α-blocker therapy, likely due to its multifactorial pathophysiology. We evaluated the efficacy and safety of desmopressin as add-on therapy for refractory nocturia in men previously treated with an α-blocker for lower urinary tract symptoms. Eligible patients were men 50 years old or older with lower urinary tract symptoms and persistent nocturia despite α-blocker treatment for a minimum of 4 weeks. The optimum dose of oral desmopressin was determined during a 4-week dose titration period and this dose was maintained for 24 weeks. Flow volume charts, International Prostate Symptom Score total and subscores, uroflowmetry and post-void residual urine volume were assessed. A total of 216 patients were enrolled in the study. Of these patients there were 158 (76%) with nocturnal polyuria, 15 (7.2%) with decreased nocturnal bladder capacity and 35 (16.8%) with nocturia due to both causes. The number of nocturnal voids significantly decreased from a baseline mean of 7.0 to 5.7 episodes for 3 days at the 24-week visit. The average International Prostate Symptom Score total and subscore significantly decreased by 4 weeks and were maintained at 24 weeks. In patients younger than 65 years, International Prostate Symptom Score voiding subscores were significantly improved at 24 weeks compared to those age 65 years or older. Desmopressin add-on therapy for refractory nocturia in men previously treated with an α-blocker for lower urinary tract symptoms improved voiding symptoms as well as nocturia, storage symptoms.


A multidimensional physical therapy program for individuals with cerebellar ataxia secondary to traumatic brain injury: a case series.
The purpose of this case series is to describe changes in impairments and activity limitations in three individuals with severe cerebellar ataxia from traumatic brain injury (TBI) who participated in a long-term, multidimensional physical therapy program. A secondary purpose is to document use of a climbing wall for these persons. Each of the individuals had a TBI, severe ataxia and was admitted to a transitional neuro-rehabilitation day treatment program. The first person, a 22-year-old, was 6 years post injury and had 127 individual physical therapy sessions over 12 months. The second person, a 16-year-old, was 5½ months post injury and had 187 individual therapy sessions over 19 months. The third person, a 20-year-old, was 6 months post injury and had 89 individual therapy sessions over 23 months. An integrative treatment approach was used, and the individuals participated in activities to minimize ataxia and improve mobility. Each of them made gains in coordination, balance, balance confidence, endurance and mobility. The three individuals with cerebellar ataxia participated in a long-term, individualized, multidimensional physical therapy treatment program, and made improvements in all areas of impairment and activity limitations. This study reinforces the need for long-term, multidimensional physical therapy for individuals with ataxia.


Improved results using OKT3 as induction immunosuppression in renal allograft recipients with delayed graft function.
Delayed graft function remains a major problem in cadaveric renal allograft transplantation. We have used 2 different immunosuppressive induction regimens in patients with delayed graft function. The first regimen, used in 40 patients from January 1985 to December 1986, consisted of CsA (8 mg/kg/day, orally within 48 hr of cadaveric renal transplantation regardless of graft function), azathioprine (1.5-2.5 mg/kg/day), and steroids (methylprednisolone 375 mg on day 0, then prednisone tapered to 30 mg/day by day 10 with slow tapering to 7.5-10 mg/day over the first 6 months after transplantation). A second regimen, used from January 1987 to March 1989, employed the same doses of azathioprine and steroids; however, OKT3 (5 mg i.v./day for 7-21 days) was administered in the 34 patients who had delayed graft function. CsA was withheld until ATN resolved. The use of OKT3 as induction immunosuppression in patients with ATN led to a significant increase in 1-year graft survival (80% vs. 55%) while markedly decreasing the incidence of rejection episodes (44% vs. 82%) and the duration of nonfunction (9.4 vs. 14.9 days). There were 5 CMV infections in patients treated with OKT3. Antibodies to OKT3 developed in only 1 of 34 patients treated with OKT3. Five of 7 patients who received a second course of OKT3 successfully reversed the rejection episode. Patient survival (89%) was the same in the 2 groups. The benefit of OKT3 on long-term graft survival appears to stem from elimination of early rejection episodes that may be difficult to diagnose in a poorly functioning allograft. We conclude that OKT3 induction provides superior results over CsA induction at doses given in renal allograft recipients with delayed graft function without a significant increase in morbidity or mortality and permits the reuse of OKT3 for treatment of rejection in most cases.


Genome-wide association study of blood pressure extremes identifies variant near UMOD associated with hypertension.
Hypertension is a heritable and major contributor to the global burden of disease. The sum of rare and common genetic variants robustly identified so far explain only 1%-2% of the population variation in BP and hypertension. This suggests the existence of more undiscovered common variants. We conducted a genome-wide association study in 1,621 hypertensive cases and 1,699 controls and follow-up validation analyses in 19,845 cases and 16,541 controls using an extreme case-control design. We identified a locus on chromosome 16 in the 5' region of Uromodulin (UMOD; rs13333226, combined P value of 3.6 × 10⁻¹¹). The minor G allele is associated with a lower risk of hypertension (OR [95%CI]: 0.87 [0.84-0.91]), reduced urinary uromodulin excretion, better renal function; and each copy of the G allele is associated with a 7.7% reduction in risk of CVD events after adjusting for age, sex, BMI, and smoking status (H.R. = 0.923, 95% CI 0.860-0.991; p = 0.027). In a subset of 13,446 individuals with estimated glomerular filtration rate (eGFR) measurements, we show that rs13333226 is independently associated with hypertension (unadjusted for eGFR: 0.89 [0.83-0.96], p = 0.004; after eGFR adjustment: 0.89 [0.83-0.96], p = 0.003). In clinical functional studies, we also consistently show the minor G allele is associated with lower urinary uromodulin excretion. The exclusive expression of uromodulin in the thick portion of the ascending limb of Henle suggests a putative role of this variant in hypertension through an effect on sodium homeostasis. The newly discovered UMOD locus for hypertension has the potential to give new insights into the role of uromodulin in BP regulation and to identify novel drugable targets for reducing cardiovascular risk.


Regularization Based Iterative Point Match Weighting for Accurate Rigid Transformation Estimation.
Feature extraction and matching (FEM) for 3D shapes finds numerous applications in computer graphics and vision for object modeling, retrieval, morphing, and recognition. However, unavoidable incorrect matches lead to inaccurate estimation of the transformation relating different datasets. Inspired by AdaBoost, this paper proposes a novel iterative re-weighting method to tackle the challenging problem of evaluating point matches established by typical FEM methods. Weights are used to indicate the degree of belief that each point match is correct. Our method has three key steps: (i) estimation of the underlying transformation using weighted least squares, (ii) penalty parameter estimation via minimization of the weighted variance of the matching errors, and (iii) weight re-estimation taking into account both matching errors and information learnt in previous iterations. A comparative study, based on real shapes captured by two laser scanners, shows that the proposed method outperforms four other state-of-the-art methods in terms of evaluating point matches between overlapping shapes established by two typical FEM methods, resulting in more accurate estimates of the underlying transformation. This improved transformation can be used to better initialize the iterative closest point algorithm and its variants, making 3D shape registration more likely to succeed.


Does prior transfusion worsen outcomes from infection in surgical patients?
Controversy continues to exist regarding the immunomodulatory effects of cellular blood transfusions in the fields of oncology, transplantation, and infectious diseases. Numerous studies have correlated transfusion with hospital-acquired infection, but the impact of transfusion on infection-related mortality has not been addressed. The objective of this study was to determine the effect of transfusion on outcomes among infected surgical patients. Data on all hospital-acquired infectious episodes among surgical intensive care unit and ward patients were collected prospectively over 39 months at a single university hospital. The relationships between prior transfusion (defined as the receipt of allogeneic red blood cells or platelets during the index hospitalization but prior to the development of infection) and over 100 variables were examined using univariate and multiple logistic regression analysis, with inclusion of a propensity score for prior transfusion to attempt to account for treatment selection bias. During the study period, 1,228 infectious episodes occurred; 641 were associated with the transfusion of packed red blood cells or platelets. Univariate analysis revealed that patients with those infectious episodes following transfusion had higher Acute Physiology and Chronic Health Evaluation II (APACHE II) scores (18.3 +/- 0.3 vs. 10.9 +/- 0.3, p < 0.0001) and higher mortality (26.0% vs. 8.9%, p < 0.0001). Those patients who died received a greater number of transfusions prior to infection than those who lived (7.7 +/- 0.8 vs. 4.5 +/- 0.3, p = 0.0002). Multiple logistic regression analysis revealed that the propensity score for prior transfusion independently predicted mortality (odds ratio 9.45, 95% confidence interval 3.71-24.09, p < 0.001), but that neither the presence or absence of transfusion (OR 0.89, 95% CI 0.54-1.46, p = 0.6) nor the absolute number of units of red blood cells or platelets transfused (OR for red blood cells 1.00, 95% CI 0.98-1.02, p = 0.7; OR for platelets 0.97, 95% CI 0.90-1.05, p = 0.5) independently predicted mortality. The transfusion of packed red blood cells or platelets prior to infection is associated with more severe disease among surgical patients, but once corrected for treatment selection bias does not appear to worsen outcomes from infection.


Sex differences in response to anti-tumor necrosis factor therapy in early and established rheumatoid arthritis -- results from the DANBIO registry.
To investigate sex differences in response to anti-tumor necrosis factor-α (TNF-α) therapy over time in early versus established rheumatoid arthritis (RA). Patients with RA who initiated anti-TNF therapy between January 2003 and June 2008 in Denmark were selected from the DANBIO Registry. Sex differences in baseline disease features were examined using chi-square, Mann-Whitney U tests, and t tests. Using a generalized estimating equations (GEE) model for repeated measures, we examined European League Against Rheumatism (EULAR) responses in men and women over 48 months of followup, adjusting for baseline values of age, 28-joint Disease Activity Score (DAS28), disease duration, and anti-TNF, methotrexate, and prednisolone use. At initiation of anti-TNF therapy (baseline), 328 women and 148 men had early RA (≤ 2 yrs), and 1245 women and 408 men had established RA (> 2 yrs). In both early and established RA, men and women had active disease with similar DAS28 scores (mean ± SD 5.2 ± 1.1), physician global scores, swollen joint counts, and radiographic changes. In early RA, men were significantly more likely to achieve a EULAR good/moderate response over 48 months compared to women (GEE; p = 0.003), and a significant interaction between sex and followup time (GEE; p < 0.0005) suggested that men achieved this response sooner than women. Better responses to anti-TNF therapy among men compared to women in early but not established RA suggest that disease duration at initiation of therapy may be an important factor to consider when investigating sex differences in treatment responses.


Neuroprotective Effects of Cornus officinalis on Stress-Induced Hippocampal Deficits in Rats and H2O2-Induced Neurotoxicity in SH-SY5Y Neuroblastoma Cells.
Oxidative stress plays a vital role in neurodegenerative diseases. Cornus officinalis (CC) has a wide range of pharmacological activities (e.g., antioxidant, neuroprotective, and anti-inflammatory). The present study was undertaken to elucidate the neuroprotective mechanism of CC and fermented CC (FCC) on stress and H2O2-induced oxidative stress damage in rats and SH-SY5Y cells. A dose of 100 mg/kg CC or FCC was orally administered to rats 1 h prior to immobilization 2 h per day for 14 days. CC, especially FCC administration decreased immobility time in forced swim test (FST), effectively alleviated the oxidative stress, and remarkably decreased corticosterone, β-endorphin and increased serotonin levels, respectively. In cells, CC and FCC significantly inhibited reactive oxygen species (ROS) generation, lactate dehydrogenase (LDH) release and significantly increased the genes expression of antioxidant and neuronal markers, such as superoxide dismutase (SOD), catalase (CAT), and brain-derived neurotrophic factor (BDNF). Moreover, the pro-apoptotic factor Bax and anti-apoptotic factor Bcl-2 (Bax/Bcl-2) ratio was regulated by CC and FCC pretreatment. Both in rats and cells, CC and FCC downregulated mitogen-activated protein kinase (MAPK) phosphorylation. Taken together, these results demonstrated that CC and particularly FCC ameliorated oxidative stress and may be used on the neuroprotection.


New natural agonists of the transient receptor potential Ankyrin 1 (TRPA1) channel.
The transient receptor potential (TRP) channels family are cationic channels involved in various physiological processes as pain, inflammation, metabolism, swallowing function, gut motility, thermoregulation or adipogenesis. In the oral cavity, TRP channels are involved in chemesthesis, the sensory chemical transduction of spicy ingredients. Among them, TRPA1 is activated by natural molecules producing pungent, tingling or irritating sensations during their consumption. TRPA1 can be activated by different chemicals found in plants or spices such as the electrophiles isothiocyanates, thiosulfinates or unsaturated aldehydes. TRPA1 has been as well associated to various physiological mechanisms like gut motility, inflammation or pain. Cinnamaldehyde, its well known potent agonist from cinnamon, is reported to impact metabolism and exert anti-obesity and anti-hyperglycemic effects. Recently, a structurally similar molecule to cinnamaldehyde, cuminaldehyde was shown to possess anti-obesity and anti-hyperglycemic effect as well. We hypothesized that both cinnamaldehyde and cuminaldehyde might exert this metabolic effects through TRPA1 activation and evaluated the impact of cuminaldehyde on TRPA1. The results presented here show that cuminaldehyde activates TRPA1 as well. Additionally, a new natural agonist of TRPA1, tiglic aldehyde, was identified and p-anisaldehyde confirmed.


Gender norms and health: insights from global survey data.
Despite global commitments to achieving gender equality and improving health and wellbeing for all, quantitative data and methods to precisely estimate the effect of gender norms on health inequities are underdeveloped. Nonetheless, existing global, national, and subnational data provide some key opportunities for testing associations between gender norms and health. Using innovative approaches to analysing proxies for gender norms, we generated evidence that gender norms impact the health of women and men across life stages, health sectors, and world regions. Six case studies showed that: (1) gender norms are complex and can intersect with other social factors to impact health over the life course; (2) early gender-normative influences by parents and peers can have multiple and differing health consequences for girls and boys; (3) non-conformity with, and transgression of, gender norms can be harmful to health, particularly when they trigger negative sanctions; and (4) the impact of gender norms on health can be context-specific, demanding care when designing effective gender-transformative health policies and programmes. Limitations of survey-based data are described that resulted in missed opportunities for investigating certain populations and domains. Recommendations for optimising and advancing research on the health impacts of gender norms are made.


Real-time, automated, interactive cardiac electrophysiology testing.
We describe a new automated interactive system that performs all of the functions required for complete evaluation of the intracardiac conduction system, for inducing, terminating and analyzing tachydysrhythmias, and for locating and characterizing accessory atrioventricular (AV) connections and ectopic foci. In the first year of operation, the system was used to conduct 210 electrophysiology studies. These ranged from simple postoperative evaluation of the conduction system to complete tachycardia studies during pharmacological manipulation of the patient. Patient age at time of catheterization ranged from 6 days to 70 years, with median age of 8 years and 7 months. Advantages accruing from automated electrophysiology testing with this system included great precision and accuracy of timing measurements, flexibility in designing and implementing pacing protocols, and rapid management of induced or spontaneous dysrhythmias. Significant savings of time and labor were achieved by computer generation of reports and graphs as opposed to manual data extraction and plotting. The single most significant advantage is the ability of the system to provide information on a real-time basis, permitting the electrophysiologist to analyze, modify, extend or curtail the procedure.


Comparative effectiveness of diagnostic testing strategies in emergency department patients with chest pain: an analysis of downstream testing, interventions, and outcomes.
Patients presenting to the emergency department (ED) with chest pain whose evaluation for ischemia demonstrates no abnormalities receive further functional or anatomical studies for coronary artery disease; however, comparative evidence for the various strategies is lacking and multiple testing options exist. To compare chest pain evaluation pathways based on their association with downstream testing, interventions, and outcomes for patients in EDs. Retrospective analysis of health insurance claims data for a national sample of privately insured patients from January 1 to December 31, 2011. Individuals with a primary or secondary diagnosis of chest pain in the ED were selected and classified into 1 of 5 testing strategies: no noninvasive testing, exercise electrocardiography, stress echocardiography, myocardial perfusion scintigraphy, or coronary computed tomography angiography. The proportion of patients in each group who received a cardiac catheterization, coronary revascularization procedure, or future noninvasive test as well as those who were hospitalized for an acute myocardial infarction (MI) during 7 and 190 days of follow-up. In 2011, there were 693 212 ED visits with a primary or secondary diagnosis of chest pain, accounting for 9.2% of all ED encounters. After application of the inclusion and exclusion criteria, 421 774 patients were included in the final analysis; 293 788 individuals did not receive an initial noninvasive test and 127 986 did, representing 1.7% of all ED encounters. Overall, the percentage of patients hospitalized with an MI was very low during both 7 and 190 days of follow-up (0.11% and 0.33%, respectively). Patients who did not undergo initial noninvasive testing were no more likely to experience an MI than were those who did receive testing. Compared with no testing, exercise electrocardiography, myocardial perfusion scintigraphy, and coronary computed tomography angiography were associated with significantly higher odds of cardiac catheterization and revascularization procedures without a concomitant improvement in the odds of experiencing an MI. Patients with chest pain evaluated in the ED who do not have an MI are at very low risk of experiencing an MI during short- and longer-term follow-up in a cohort of privately insured patients. This low risk does not appear to be affected by the initial testing strategy. Deferral of early noninvasive testing appears to be reasonable.


Relationship between hexokinase and the aquaporin PIP1 in the regulation of photosynthesis and plant growth.
Increased expression of the aquaporin NtAQP1, which is known to function as a plasmalemma channel for CO₂ and water, increases the rate of both photosynthesis and transpiration. In contrast, increased expression of Arabidopsis hexokinase1 (AtHXK1), a dual-function enzyme that mediates sugar sensing, decreases the expression of photosynthetic genes and the rate of transpiration and inhibits growth. Here, we show that AtHXK1 also decreases root and stem hydraulic conductivity and leaf mesophyll CO₂ conductance (g(m)). Due to their opposite effects on plant development and physiology, we examined the relationship between NtAQP1 and AtHXK1 at the whole-plant level using transgenic tomato plants expressing both genes simultaneously. NtAQP1 significantly improved growth and increased the transpiration rates of AtHXK1-expressing plants. Reciprocal grafting experiments indicated that this complementation occurs when both genes are expressed simultaneously in the shoot. Yet, NtAQP1 had only a marginal effect on the hydraulic conductivity of the double-transgenic plants, suggesting that the complementary effect of NtAQP1 is unrelated to shoot water transport. Rather, NtAQP1 significantly increased leaf mesophyll CO₂ conductance and enhanced the rate of photosynthesis, suggesting that NtAQP1 facilitated the growth of the double-transgenic plants by enhancing mesophyll conductance of CO₂.


Op HERRICK primary care casualties: the forgotten many.
The number of battle casualties generated during war is far outnumbered by non-battle casualties. Each year the current conflict in Afghanistan sees hundreds of service personnel medically evacuated direct from the front line to the care of their home units' primary care facility. To date these casualties remain undiscovered by medical research. This is the first study to look at the care pathway of primary care casualties from Operation HERRICK using information from the Defence Patient Tracking System (DPTS). Information relating to all casualties from Afghanistan discharged at the airhead between 1 January 2009 and 31 December 2010 was collected from the DPTS. Common conditions were identified and information relating to the follow up care extracted to provide an overview of the care pathway. 387 aeromedical evacuations were identified as primary care casualties. The three commonest conditions were musculoskeletal (183 cases), mental health (29) and noise-induced hearing loss (26). 205 (53%) were not seen outside of primary care for the tracked condition. 166 (81%) of those that remained under primary care had two or less consultations during the time period of the study. The mean time frame between the 1st and 2nd consultation was 5.4 weeks. A significant number of aeromedical evacuations from Afghanistan are for primary care casualties. The DPTS can be used to provide a basic overview of the care pathway of repatriated personnel. Little contact with the medical services would appear to occur for these types of casualties. There is a significant gap in military medical research looking at primary care casualties repatriated from operations.


Tensions in support for family caregivers of people with dementia in Singapore: A qualitative study.
Family members as informal caregivers are considered the first line of support for people with dementia across the world. In Singapore, caregiving expectations revolve around the cultural expectations of providing care in the home environment. However, studies in Singapore have identified a lack of family support for primary caregivers. Family support has been discussed in the literature as the provision of care for people with dementia, and rarely as a resource for family caregivers. To understand family support among primary caregivers in Singapore, 24 semi-structured interviews were conducted. Thematic analysis found four themes: excuses for lack of physical support for the caregiver, tensions between cultural expectations of caregiving and the provision of support, unmet emotional support, and lack of awareness of dementia and caregiving needs. Caregivers rationalized and forgave the absence of physical support but were frustrated when the lack of support impacted people with dementia. This was seen as a lack of fulfilling cultural obligations of caring for elderly parents. The caregivers also felt frustrated with the lack of emotional support provided to them, but these were unspoken between the caregiver and the family members. Insufficient and unhelpful support giving was exacerbated with the perception of family members' limited understanding of the demands of caregiving. The findings offer four practical suggestions to address unmet support needs. First, public education is needed to enhance general knowledge about the symptoms and progression of dementia. Second, help is needed to address miscommunication about support within the family. Third, the development of guidebooks is needed to help family caregivers communicate with family members about their various support needs. Fourth, the relationship between cultural expectation and caregiving must be understood within the context of modernity and urbanism.


HER2 oncogenic function escapes EGFR tyrosine kinase inhibitors via activation of alternative HER receptors in breast cancer cells.
The response rate to EGFR tyrosine kinase inhibitors (TKIs) may be poor and unpredictable in cancer patients with EGFR expression itself being an inadequate response indicator. There is limited understanding of the mechanisms underlying this resistance. Furthermore, although TKIs suppress the growth of HER2-overexpressing breast tumor cells, they do not fully inhibit HER2 oncogenic function at physiological doses. Here we have provided a molecular mechanism of how HER2 oncogenic function escapes TKIs' inhibition via alternative HER receptor activation as a result of autocrine ligand release. Using both Förster Resonance Energy Transfer (FRET) which monitors in situ HER receptor phosphorylation as well as classical biochemical analysis, we have shown that the specific tyrosine kinase inhibitors (TKIs) of EGFR, AG1478 and Iressa (Gefitinib) decreased EGFR and HER3 phosphorylation through the inhibition of EGFR/HER3 dimerization. Consequent to this, we demonstrate that cleavage of HER4 and dimerization of HER4/HER2 occur together with reactivation of HER3 via HER2/HER3, leading to persistent HER2 phosphorylation in the now resistant, surviving cells. These drug treatment-induced processes were found to be mediated by the release of ligands including heregulin and betacellulin that activate HER3 and HER4 via HER2. Whereas an anti-betacellulin antibody in combination with Iressa increased the anti-proliferative effect in resistant cells, ligands such as heregulin and betacellulin rendered sensitive SKBR3 cells resistant to Iressa. These results demonstrate the role of drug-induced autocrine events leading to the activation of alternative HER receptors in maintaining HER2 phosphorylation and in mediating resistance to EGFR tyrosine kinase inhibitors (TKIs) in breast cancer cells, and hence specify treatment opportunities to overcome resistance in patients.


The adipokinetic hormone system in Culicinae (Diptera: Culicidae): molecular identification and characterization of two adipokinetic hormone (AKH) precursors from Aedes aegypti and Culex pipiens and two putative AKH receptor variants from A. aegypti.
Insect neuropeptides of the adipokinetic hormone (AKH) family induce the mobilization of energy stores to fuel flight, but also affect the nutritional balance during diapause and oogenesis. They are therefore important regulators for flight, hibernation, and reproduction in mosquitoes including those that transmit human pathogens. In this study, we identified and analyzed the genes encoding two AKH preprohormones in the Yellow fever mosquito, Aedes aegypti: Aedae-AKH-I encodes the octapeptide pELFTPSWa and Aedae-AKH-II the decapeptide pEVTFSRDWNAa. Identical AKHs were identified in the West Nile virus vector, Culex pipiens, whose genes were characterized in this study as Culpi-AKH-I and Culpi-AKH-II. Using Northern blot, transcript expression was shown in A. aegypti, for Aedae-AKH-I in the head/thorax tissues of pupae and females, as well as in the abdomen of adult males; Aedae-AKH-II was only expressed in adults. In an immunocytological study using an AKH-antibody, the corpus cardiacum (CC), the intrinsic CC-cells (X-cells), the nervi corporis cardiaci, cells in the brain and thoracic ganglia were stained. In addition, two splice variants of the AKH-receptor gene were characterized in A. aegypti, (Aedae-AKHR-I and -II). RT-PCR revealed that both variants of these typical G-protein-coupled receptors were expressed in all life stages. Aedae-AKHR-I expression was also detected in the ovaries, indicating once more the influence of the AKH/AKHR system during the insect's oogenesis. Based on phylogenetic data, we postulate two closely related types of AKH-receptors that could bind selectively the two AKH peptides found in A. aegypti.


Why do clinicians sometimes find it difficult to use the results of systematic reviews in routine clinical practice?
Reviews of systematic results have proved enormously useful in routine clinical practice. However, the uncritical application of the overall results of systematic reviews to clinical decisions about individual patients can sometimes do more harm than good. Clinical syndromes are often complex, with considerable pathological heterogeneity, and the likely balance of the risks and benefits of interventions can be difficult to judge on an individual patient basis. Active involvement of experienced clinicians is essential in the planning, reporting, and interpretation of reviews. Detailed consideration should be given to the possibility of diagnostic or pathological heterogeneity within the clinical trials included in the review or between the trial patients and patients seen in routine clinical practice. For treatments that have significant risk of harm or treatments that are particularly costly, meta-analysis of individual patient data with stratification by baseline risk is sometimes helpful in targeting treatment appropriately in routine clinical practice.


Mechanical characterisation of the lateral collateral ligament complex of the ankle at realistic sprain-like strain rates.
Synthetic interventions continue to evolve with the progression made in materials science, surgical technologies and surgical methods. To facilitate the evolution of synthetic devices for lateral ankle repair a better understanding of the mechanical properties and failure mechanisms of the lateral collateral ligament (LCL) complex is required. This study aimed to improve understanding of the mechanical properties and failure modes of the LCL complex at strain rates representative of sprain. The LCLs were dissected from six human cadavers to produce individual bone-ligament-bone specimens. A mechanical testing device uni-axially loaded the ligaments in tension. Initially, preconditioning between 2 N and a load value corresponding to 3.5% strain was conducted for 15 cycles, before extension to failure at strain rate of 100%.s-1. The results were stratified by age, weight and body mass index (BMI) to explore potential correlations with ligament ultimate failure load or ligament stiffness. The mean ultimate failure loads and the 95% confidence intervals for the ATFL, calcaneofibular (CFL) and posterior talofibular (PTFL) ligaments were 263.6 ± 164.3 N, 367.8 ± 79.8 N and 351.4 ± 110.8 N, respectively. A strong positive Pearson correlation was found between BMI and ultimate failure load of the CFL (r = 0.919; P = .01). A non-significant relationship was found between the mechanical properties and both age and weight. The ATFL avulsed from the fibula four times, the CFL avulsed from the fibula twice, the PTFL avulsed from the talus twice and all remaining failures were mid-substance. The results identify the forces required to induce failure of the individual ligaments of the LCL complex and the related failure modes of individual ligaments. A correlation may exist between BMI and the ultimate failure load of the CFL and PTFL, although a greater sample size is required for confirmation.


Modelling Emergence of Wolbachia Toxin-Antidote Protein Functions with an Evolutionary Algorithm.
Evolutionary algorithms (EAs) simulate Darwinian evolution and adeptly mimic natural evolution. Most EA applications in biology encode high levels of abstraction in top-down ecological population models. In contrast, our research merges protein alignment algorithms from bioinformatics into codon based EAs that simulate molecular protein string evolution from the bottom up. We apply our EA to reconcile a problem in the field of Wolbachia induced cytoplasmic incompatibility (CI). Wolbachia is a microbial endosymbiont that lives inside insect cells. CI is conditional insect sterility that operates as a toxin antidote (TA) system. Although, CI exhibits complex phenotypes not fully explained under a single discrete model. We instantiate in-silico genes that control CI, CI factors ( cifs ), as strings within the EA chromosome. We monitor the evolution of their enzymatic activity, binding, and cellular localization by applying selective pressure on their primary amino acid strings. Our model helps rationalize why two distinct mechanisms of CI induction might coexist in nature. We find that nuclear localization signals (NLS) and Type IV secretion system signals (T4SS) are of low complexity and evolve fast, whereas binding interactions have intermediate complexity, and enzymatic activity is the most complex. Our model predicts that as ancestral TA systems evolve into eukaryotic CI systems, the placement of NLS or T4SS signals can stochastically vary, imparting effects that might impact CI induction mechanics. Our model highlights how preconditions, genetic diversity, and sequence length can bias evolution of cifs towards one mechanism or another.


Comparison of thiol/disulphide homeostasis parameters in patients with COPD, asthma and ACOS.
Chronic obstructive pulmonary disease (COPD), asthma and asthma-COPD overlap syndrome (ACOS) are obstructive pulmonary disorders with different manifestations. Status of oxidation in tissues is important in obstructive pulmonary disorders. Smoking, acute exacerbations of COPD and asthma were associated with a marked imbalance in oxidant or antioxidant status due to increased oxidative stress in tissues and blood. Oxidative conditions may cause a reversible formation of mixed disulphides among protein thiol groups. The aim of this study was to compare parameters related with thiol/disulphide homeostasis in patients with COPD, asthma and ACOS. Patients (n= 135, 69 females, 66 males) who were referred with a diagnosis of COPD, asthma or ACOS were included in the study. Thiol/ disulphide homeostasis parameters in blood were analysed by a newly established method that measures the exact thiol/ disulphide status in the body. The patients with COPD, asthma or ACOS were similar for demographic parameters other than age and number of cigarettes smoked. Measured thiol/disulphide homeostasis parameters were similar among these patient groups. When these biochemical measurements were adjusted for age and number of cigarettes by using regression analysis, similarity for thiol/disulphide homeostasis parameters among patient groups persisted. To best of our knowledge, this is the first study to compare thiol/disulphide homeostasis parameters in COPD, asthma and ACOS patients. Similarity of thiol/disulphide homeostasis parameters among these patient groups supports the current view of Dutch hypothesis that COPD, asthma and ACOS share similar pathophysiological features but display different clinical manifestations.


The concentrations of five heavy metals in components of an economically important urban coastal wetland in Ghana: public health and phytoremediation implications.
Sakumo II is an urban wetland and a receptacle for domestic and industrial wastes from two cities in Ghana. It however supports viable populations of fish and crabs, is cultivated for food crops and grazed by farm animals. Components of the wetland can therefore accumulate pollutants, but the public health and phytoremediation implications of this are yet to be evaluated. We analysed Cd, As, Hg, Cu and Pb in the lagoon water, sediment, green algae, eight species of aquatic macrophytes, seven species of arthropods and one species of fish. The concentrations of Pb were generally below detection limit whilst Cu was detected only in the lagoon water and Pheropsophus vertialis. Cadmium ranged from 21 ± 4 ppb in algae to 69 ± 12 ppb in Typha domingensis and was generally higher than As and Hg. The highest concentration of As was 11.7 ± 2.1 ppb in Pistia stratiotes whilst Hg was highest in lagoon water (4 ± 2 ppb). The Cd concentrations generally, and Hg concentrations in macrophytes, were higher than US EPA guidelines indicating the wetland's resources were unsafe for regular consumption. Among the emergent aquatic macrophytes, T. domingensis, Ludwigia sp. and Paspalum vaginatum, respectively, had the highest accumulation capacity for Cd, As and Hg, but the floating aquatic plant P. stratiotes appeared to be a better accumulator of Cd and As.


Inhibition of Ca(2+) signalling by p130, a phospholipase-C-related catalytically inactive protein: critical role of the p130 pleckstrin homology domain.
p130 was originally identified as an Ins(1,4,5)P(3)-binding protein similar to phospholipase C-delta but lacking any phospholipase activity. In the present study we have further analysed the interactions of p130 with inositol compounds in vitro. To determine which of the potential ligands interacts with p130 in cells, we performed an analysis of the cellular localization of this protein, the isolation of a protein-ligand complex from cell lysates and studied the effects of p130 on Ins(1,4,5)P(3)-mediated Ca(2+) signalling by using permeabilized and transiently or stably transfected COS-1 cells (COS-1(p130)). In vitro, p130 bound Ins(1,4,5)P(3) with a higher affinity than that for phosphoinositides. When the protein was isolated from COS-1(p130) cells by immunoprecipitation, it was found to be associated with Ins(1,4,5)P(3). Localization studies demonstrated the presence of the full-length p130 in the cytoplasm of living cells, not at the plasma membrane. In cell-based assays, p130 had an inhibitory effect on Ca(2+) signalling. When fura-2-loaded COS-1(p130) cells were stimulated with bradykinin, epidermal growth factor or ATP, it was found that the agonist-induced increase in free Ca(2+) concentration, observed in control cells, was inhibited in COS-1(p130). This inhibition was not accompanied by the decreased production of Ins(1,4,5)P(3); the intact p130 pleckstrin homology domain, known to be the ligand-binding site in vitro, was required for this effect in cells. These results suggest that Ins(1,4,5)P(3) could be the main p130 ligand in cells and that this binding has the potential to inhibit Ins(1,4,5)P(3)-mediated Ca(2+) signalling.


Effect of ketone infusions on amino acid and nitrogen metabolism in man.
To evaluate the role of hyperketonemia in the hypoalaninemia and decreased protein catabolism of prolonged starvation, Na dl-beta-hydroxybutyrate was administered as a primed continuous 3-6-h infusion in nonobese subjects and in obese subjects in the postabsorptive state and after 3 days and 3-5 1/2 wk of starvation. An additional obese group received 12-h ketone infusions on 2 consecutive days after 5-10 wk of fasting. The ketone infusion in nonobese and obese subjects studied in the postabsorptive state resulted in total blood ketone acid levels of 1.1-1.2 mM, a 5-15 mg/100 ml decrease in plasma glucose, and unchanged levels of insulin, glucagon, lactate, and pyruvate. Plasma alanine fell by 21% (P smaller than 0.001) in 3 h. In contrast, other amino acids were stable or varied by less than 10%. Infusions lasting 6 h reduced plasma alanine by 37%, reaching levels comparable to those observed in prolonged starvation. Equimolar infusions of NaC1 and/or administration of NaHCO3 failed to alter plasma alanine levels. During prolonged fasting, plasma alanine, which had fallen by 40% below prefast levels, fell an additional 30% in response to the ketone infusion. In association with repeated prolonged (12 h) infusions in subjects fasted 5-10 wk, urinary nitrogen excretion fell by 30%, returning to base line after cessation of theinfusions and paralleling the changes in plasma alanine. Ketone infusins resulted in two- to fourfold greater increments in blood ketone acids in fasted as compared to postabsorptive subjects. It is concluded that increased blood ketone acid levels induced by infusions of Na DL-beta-hydroxybutyrate result in hypoalaninemia and in nitrogen conservation in starvation. These data suggest that hyperketonemia may be a contributory factor in the decreased availability or circulating alanine and reduction in protein catabolism characteristic of prolonged fastings9


Low plasma haptoglobin is a risk factor for life-threatening childhood severe malarial anemia and not an exclusive consequence of hemolysis.
Severe Malarial Anemia (SMA), a life-threatening childhood Plasmodium falciparum malaria syndrome requiring urgent blood transfusion, exhibits inflammatory and hemolytic pathology. Differentiating between hypo-haptoglobinemia due to hemolysis or that of genetic origin is key to understand SMA pathogenesis. We hypothesized that while malaria-induced hypo-haptoglobinemia should reverse at recovery, that of genetic etiology should not. We carried-out a case-control study of children living under hyper-endemic holoendemic malaria burden in the sub-Saharan metropolis of Ibadan, Nigeria. We show that hypo-haptoglobinemia is a risk factor for childhood SMA and not solely due to intravascular hemolysis from underlying schizogony. In children presenting with SMA, hypo-haptoglobinemia remains through convalescence to recovery suggesting a genetic cause. We identified a haptoglobin gene variant, rs12162087 (g.-1203G > A, frequency = 0.67), to be associated with plasma haptoglobin levels (p = 8.5 × 10-6). The Homo-Var:(AA) is associated with high plasma haptoglobin while the reference Homo-Ref:(GG) is associated with hypo-haptoglobinemia (p = 2.3 × 10-6). The variant is associated with SMA, with the most support for a risk effect for Homo-Ref genotype. Our insights on regulatory haptoglobin genotypes and hypo-haptoglobinemia suggest that haptoglobin screening could be part of risk-assessment algorithms to prevent rapid disease progression towards SMA in regions with no-access to urgent blood transfusion where SMA accounts for high childhood mortality rates.


The structure, splicing, synteny and expression of lamprey COE genes and the evolution of the COE gene family in chordates.
COE genes encode transcription factors that have been found in all metazoans examined to date. They possess a distinctive domain structure that includes a DNA-binding domain (DBD), an IPT/TIG domain and a helix-loop-helix (HLH) domain. An intriguing feature of the COE HLH domain is that in jawed vertebrates it is composed of three helices, compared to two in invertebrates. We report the isolation and expression of two COE genes from the brook lamprey Lampetra planeri and compare these to COE genes from the lampreys Lethenteron japonicum and Petromyzon marinus. Molecular phylogenetic analyses do not resolve the relationship of lamprey COE genes to jawed vertebrate paralogues, though synteny mapping shows that they all derive from duplication of a common ancestral genomic region. All lamprey genes encode conserved DBD, IPT/TIG and HLH domains; however, the HLH domain of lamprey COE-A genes encodes only two helices while COE-B encodes three helices. We also identified COE-B splice variants encoding either two or three helices in the HLH domain, along with other COE-A and COE-B splice variants affecting the DBD and C-terminal transactivation regions. In situ hybridisation revealed expression in the lamprey nervous system including the brain, spinal cord and cranial sensory ganglia. We also detected expression of both genes in mesenchyme in the pharyngeal arches and underlying the notochord. This allows us to establish the primitive vertebrate expression pattern for COE genes and compare this to that of invertebrate chordates and other animals to develop a model for COE gene evolution in chordates.


Increased Equivalent Input Noise in Glaucomatous Central Vision: Is it Due to Undersampling of Retinal Ganglion Cells?
Recent evidence shows that macular damage is common even in early stages of glaucoma. Here we investigated whether contrast sensitivity loss in the central vision of glaucoma patients is due to an increase in equivalent input noise (Neq), a decrease in calculation efficiency, or both. We also examined how retinal undersampling resulting from loss of retinal ganglion cells (RGCs) may affect Neq and calculation efficiency. This study included 21 glaucoma patients and 23 age-matched normally sighted individuals. Threshold contrast for orientation discrimination was measured with a sinewave grating embedded in varying levels of external noise. Data were fitted to the linear amplifier model (LAM) to factor contrast sensitivity into Neq and calculation efficiency. We also correlated macular RGC counts estimated from structural (spectral-domain optical coherence tomography) and functional (standard automated perimetry Swedish interactive thresholding algorithm 10-2) data with either Neq or efficiency. Furthermore, using analytical and computer simulation approach, the relative effect of retinal undersampling on Neq and efficiency was evaluated by adding the RGC sampling module into the LAM. Compared with normal controls, glaucoma patients exhibited a significantly larger Neq without significant difference in efficiency. Neq was significantly correlated with Pelli-Robson contrast sensitivity and macular RGC counts. The results from analytical derivation and model simulation further demonstrated that Neq can be expressed as a function of internal noise and retinal sampling. Our results showed that equivalent input noise is significantly elevated in glaucomatous vision, thereby impairing foveal contrast sensitivity. Our findings further elucidated how undersampling at the retinal level may increase equivalent input noise.


Induction of neostriatal neurogenesis slows disease progression in a transgenic murine model of Huntington disease.
Ependymal overexpression of brain-derived neurotrophic factor (BDNF) stimulates neuronal addition to the adult striatum, from subependymal progenitor cells. Noggin, by suppressing subependymal gliogenesis and increasing progenitor availability, potentiates this process. We asked whether BDNF/Noggin overexpression might be used to recruit new striatal neurons in R6/2 huntingtin transgenic mice. R6/2 mice injected with adenoviral BDNF and adenoviral Noggin (AdBDNF/AdNoggin) recruited BrdU(+)betaIII-tubulin(+) neurons, which developed as DARPP-32(+) and GABAergic medium spiny neurons that expressed either enkephalin or substance P and extended fibers to the globus pallidus. Only AdBDNF/AdNoggin-treated R6/2 mice harbored migrating doublecortin-defined neuroblasts in their striata, and the new neurons expressed p27 as a marker of mitotic quiescence after parenchymal integration. AdBDNF/AdNoggin-treated R6/2 mice sustained their rotarod performance and open-field activity and survived longer than did AdNull-treated and untreated controls. Neither motor performance nor survival improved in R6/2 mice treated only with AdBDNF, and intraventricular infusion of the mitotic inhibitor Ara-C completely blocked the performance and survival effects of AdBDNF/AdNoggin, suggesting that the benefits of AdBDNF/AdNoggin derived from neuronal addition. Thus, BDNF and Noggin induced striatal neuronal regeneration, delayed motor impairment, and extended survival in R6/2 mice, suggesting a new therapeutic strategy in Huntington disease.


Modulation of Rac1 activity by ADMA/DDAH regulates pulmonary endothelial barrier function.
Endogenously produced nitric oxide synthase inhibitor, asymmetric methylarginine (ADMA) is associated with vascular dysfunction and endothelial leakage. We studied the role of ADMA, and the enzymes metabolizing it, dimethylarginine dimethylaminohydrolases (DDAH) in the regulation of endothelial barrier function in pulmonary macrovascular and microvascular cells in vitro and in lungs of genetically modified heterozygous DDAHI knockout mice in vivo. We show that ADMA increases pulmonary endothelial permeability in vitro and in in vivo and that this effect is mediated by nitric oxide (NO) acting via protein kinase G (PKG) and independent of reactive oxygen species formation. ADMA-induced remodeling of actin cytoskeleton and intercellular adherens junctions results from a decrease in PKG-mediated phosphorylation of vasodilator-stimulated phosphoprotein (VASP) and a subsequent down-regulation of Rac1 activity. The effects of ADMA on endothelial permeability, Rac1 activation and VASP phosphorylation are prevented by overexpression of active DDAHI and DDAHII, whereas inactive DDAH mutants have no effect. These findings demonstrate for the first time that ADMA metabolism critically determines pulmonary endothelial barrier function by modulating Rac1-mediated remodeling of the actin cytoskeleton and intercellular junctions.


An inulin-type fructan enhances calcium absorption primarily via an effect on colonic absorption in humans.
Calcium absorption efficiency and bone mineral mass are increased in adolescents who regularly consume inulin-type fructans (ITF). The mechanism of action in increasing absorption is unknown but may be related to increased colonic calcium absorption. We conducted a study in young adults designed to evaluate these mechanisms with a kinetic technique using (42)Ca orally and (46)Ca dosed i.v. Those who responded to 8 wk of supplementation with 8 g of a mixed short and long degree of polymerization ITF by increasing their calcium absorption had kinetic measurements analyzed to evaluate the time course of absorption. The area under the curve of the oral tracer in the blood during the 26 h after dosing was calculated and the time dependence of increased absorption determined. Eight young adults (of 13 studied), with mean calcium intake approximately 900 mg/d, responded to the ITF with an increased calcium absorption of at least 3%. In responders, absorption increased from 22.7 +/- 11.3% to 31.0 +/- 15.3%. Colonic absorption, defined as absorption that occurred >7 h after oral dosing, represented 69.6 +/- 18.6% of the increase, or 49 +/- 28 mg/d. These findings suggest that, in those who respond to ITF, its effects on calcium absorption occur principally in the colon. This benefit to ITF may be especially important when absorption in the small intestine is impaired for anatomic or physiological reasons.


Differential effects of resistance and endurance exercise in the fed state on signalling molecule phosphorylation and protein synthesis in human muscle.
Resistance (RE) and endurance (EE) exercise stimulate mixed skeletal muscle protein synthesis. The phenotypes induced by RE (myofibrillar protein accretion) and EE (mitochondrial expansion) training must result from differential stimulation of myofibrillar and mitochondrial protein synthesis. We measured the synthetic rates of myofibrillar and mitochondrial proteins and the activation of signalling proteins (Akt-mTOR-p70S6K) at rest and after an acute bout of RE or EE in the untrained state and after 10 weeks of RE or EE training in young healthy men. While untrained, RE stimulated both myofibrillar and mitochondrial protein synthesis, 67% and 69% (P < 0.02), respectively. After training, only myofibrillar protein synthesis increased with RE (36%, P = 0.05). EE stimulated mitochondrial protein synthesis in both the untrained, 154%, and trained, 105% (both P < 0.05), but not myofibrillar protein synthesis. Acute RE and EE increased the phosphorylation of proteins in the Akt-mTOR-p70S6K pathway with comparatively minor differences between two exercise stimuli. Phosphorylation of Akt-mTOR-p70S6K proteins was increased after 10 weeks of RE training but not by EE training. Chronic RE or EE training modifies the protein synthetic response of functional protein fractions, with a shift toward exercise phenotype-specific responses, without an obvious explanatory change in the phosphorylation of regulatory signalling pathway proteins.


Isomer-Pure Bis-PCBM-Assisted Crystal Engineering of Perovskite Solar Cells Showing Excellent Efficiency and Stability.
A fullerene derivative (α-bis-PCBM) is purified from an as-produced bis-phenyl-C61 -butyric acid methyl ester (bis-[60]PCBM) isomer mixture by preparative peak-recycling, high-performance liquid chromatography, and is employed as a templating agent for solution processing of metal halide perovskite films via an antisolvent method. The resulting α-bis-PCBM-containing perovskite solar cells achieve better stability, efficiency, and reproducibility when compared with analogous cells containing PCBM. α-bis-PCBM fills the vacancies and grain boundaries of the perovskite film, enhancing the crystallization of perovskites and addressing the issue of slow electron extraction. In addition, α-bis-PCBM resists the ingression of moisture and passivates voids or pinholes generated in the hole-transporting layer. As a result, a power conversion efficiency (PCE) of 20.8% is obtained, compared with 19.9% by PCBM, and is accompanied by excellent stability under heat and simulated sunlight. The PCE of unsealed devices dropped by less than 10% in ambient air (40% RH) after 44 d at 65 °C, and by 4% after 600 h under continuous full-sun illumination and maximum power point tracking, respectively.


Muscle oxygenation, EMG, and cardiovascular responses for cabin attendants vs. controls.
The goal was to investigate the effect of acute moderate hypobaric exposure on the physiological responses to sustained contractions (local) and light to moderate dynamic exercise (systemic) for cabin attendants (CAB) and a matched control group (CON). There were 14 CAB and 13 CON who participated. The experimental protocol was designed as a 2 x 2 randomized crossover study for which subjects performed a forearm isometric test and a cycling test in normobaric (NORM) and hypobaric [HYPO; 574 mmHg approximately 8000 ft (2440 m) above sea level (ASL)] conditions. Oxygen saturation (StO2%) was measured in the extensor carpi radialis muscle at rest and during an isometric wrist extension (20% MVC). Heart rate (HR), ventilation (V(E)), and oxygen uptake (VO2) were measured at rest, during cycling exercise at intensities of 50 W and 100 W, and during recovery. Blood samples were collected and analyzed for hemoglobin concentrations. Local response: There was a significant effect of HYPO for both CAB and CON with a decrease in StO2% of 6.5% both at rest and during contraction. Systemic response: there was no general effect of HYPO on HR, VE, or VO2. No differences were found between groups in either the local or systemic response or in the blood parameters. This study shows an effect of moderate hypobaric exposure on local muscle oxygenation during rest and sustained contraction. Whether this has an impact on fatigue development during work among cabin attendants is discussed.


Metabolic characteristics of human adipose tissue in vivo.
1. A method was developed for sampling the venous drainage from the subcutaneous adipose tissue of the anterior abdominal wall. This is a large depot in many subjects, and seems well suited to such studies as it is completely separated from the venous drainage of the underlying muscle by the aponeurosis of the external oblique muscle. 2. Eight normal subjects were studied after an overnight fast, and for 120 min after ingestion of 75 g of glucose. Concentrations of substrates in the abdominal wall drainage were compared with those in arterialized blood and in forearm muscle drainage. 3. Non-esterified fatty acid and glycerol concentrations in the abdominal wall drainage were high (three to four times the arterial level) after overnight fast. After glucose ingestion, arterial and abdominal venous levels fell and the arteriovenous differences narrowed. The forearm showed uptake of non-esterified fatty acids when fasting but not after glucose ingestion, with no significant arteriovenous difference for glycerol at any time. 4. The abdominal wall tissues showed a small arteriovenous difference for glucose uptake during fasting, which increased after glucose ingestion. Although lactate was produced throughout, its molar ratio to glucose uptake was less than that reported for other superficial sites, suggesting only a minor contribution of skin metabolism. Forearm muscle showed a larger and more prolonged increase in arteriovenous difference for glucose uptake after the glucose load, but no consistent release or uptake of lactate. 5. We conclude that the tissue studied by this technique is predominantly adipose. This technique may have wide application in studies of the metabolic basis for body weight regulation in man.


Are There Trigger Points in the Spastic Muscles? Electromyographical Evidence of Dry Needling Effects on Spastic Finger Flexors in Chronic Stroke.
The purpose was to examine the immediate effects of dry needling to spastic finger muscles in chronic stroke. Ten chronic stroke patients with spasticity in finger flexors participated in this experiment. Dry needling to the flexor digitorum superficialis (FDS) muscle was performed under ultrasound guidance for about 30 s (about 100 times). Clinical assessment and intramuscular needle EMG readings were made before and immediately after dry needling. Immediately after needling, the FDS muscle was felt less tight to palpation and the proximal phalangeal joint rested in a less flexed position (p = 0.036). The MAS score decreased for FDS (p = 0.017) and flexor digitorum profundus (FDP) (p = 0.029). Motor unit action potential (MUAP) spikes decreased from 41.6 ± 5.5 to 6.7 ± 2.2 spikes/s (p = 0.002), an 84% reduction after dry needling. However, the pre-needling spike frequency was not correlated to MAS or resting position of the FDS muscles. Dry needling to the spastic finger flexors leads to immediate spasticity reduction, increased active range of motion, and decreased frequency of motor unit spontaneous firing spikes. The results suggest that latent trigger points possibly exist in spastic muscles and they contribute partly to spastic hypertonia of finger flexors in chronic stroke.


Identification of common variants associated with human hippocampal and intracranial volumes.
Identifying genetic variants influencing human brain structures may reveal new biological mechanisms underlying cognition and neuropsychiatric illness. The volume of the hippocampus is a biomarker of incipient Alzheimer's disease and is reduced in schizophrenia, major depression and mesial temporal lobe epilepsy. Whereas many brain imaging phenotypes are highly heritable, identifying and replicating genetic influences has been difficult, as small effects and the high costs of magnetic resonance imaging (MRI) have led to underpowered studies. Here we report genome-wide association meta-analyses and replication for mean bilateral hippocampal, total brain and intracranial volumes from a large multinational consortium. The intergenic variant rs7294919 was associated with hippocampal volume (12q24.22; N = 21,151; P = 6.70 × 10(-16)) and the expression levels of the positional candidate gene TESC in brain tissue. Additionally, rs10784502, located within HMGA2, was associated with intracranial volume (12q14.3; N = 15,782; P = 1.12 × 10(-12)). We also identified a suggestive association with total brain volume at rs10494373 within DDR2 (1q23.3; N = 6,500; P = 5.81 × 10(-7)).


Slow axial drift in three-dimensional granular tumbler flow.
Models of monodisperse particle flow in partially filled three-dimensional tumblers often assume that flow along the axis of rotation is negligible. We test this assumption, for spherical and double cone tumblers, using experiments and discrete element method simulations. Cross sections through the particle bed of a spherical tumbler show that, after a few rotations, a colored band of particles initially perpendicular to the axis of rotation deforms: particles near the surface drift toward the pole, while particles deeper in the flowing layer drift toward the equator. Tracking of mm-sized surface particles in tumblers with diameters of 8-14 cm shows particle axial displacements of one to two particle diameters, corresponding to axial drift that is 1-3% of the tumbler diameter, per pass through the flowing layer. The surface axial drift in both double cone and spherical tumblers is zero at the equator, increases moving away from the equator, and then decreases near the poles. Comparing results for the two tumbler geometries shows that wall slope causes axial drift, while drift speed increases with equatorial diameter. The dependence of axial drift on axial position for each tumbler geometry is similar when both are normalized by their respective maximum values.


Exhaustion of food budgets at month's end and hospital admissions for hypoglycemia.
One in seven US households cannot reliably afford food. Food budgets are more frequently exhausted at the end of a month than at other points in time. We postulated that this monthly pattern influenced health outcomes, such as risk for hypoglycemia among people with diabetes. Using administrative data on inpatient admissions in California for 2000-08, we found that admissions for hypoglycemia were more common in the low-income than the high-income population (270 versus 200 admissions per 100,000). Risk for hypoglycemia admission increased 27 percent in the last week of the month compared to the first week in the low-income population, but we observed no similar temporal variation in the high-income population. These findings suggest that exhaustion of food budgets might be an important driver of health inequities. Policy solutions to improve stable access to nutrition in low-income populations and raise awareness of the health risks of food insecurity might be warranted.


Genetic Disruption of Circadian Rhythms in the Suprachiasmatic Nucleus Causes Helplessness, Behavioral Despair, and Anxiety-like Behavior in Mice.
Major depressive disorder is associated with disturbed circadian rhythms. To investigate the causal relationship between mood disorders and circadian clock disruption, previous studies in animal models have employed light/dark manipulations, global mutations of clock genes, or brain area lesions. However, light can impact mood by noncircadian mechanisms; clock genes have pleiotropic, clock-independent functions; and brain lesions not only disrupt cellular circadian rhythms but also destroy cells and eliminate important neuronal connections, including light reception pathways. Thus, a definitive causal role for functioning circadian clocks in mood regulation has not been established. We stereotactically injected viral vectors encoding short hairpin RNA to knock down expression of the essential clock gene Bmal1 into the brain's master circadian pacemaker, the suprachiasmatic nucleus (SCN). In these SCN-specific Bmal1-knockdown (SCN-Bmal1-KD) mice, circadian rhythms were greatly attenuated in the SCN, while the mice were maintained in a standard light/dark cycle, SCN neurons remained intact, and neuronal connections were undisturbed, including photic inputs. In the learned helplessness paradigm, the SCN-Bmal1-KD mice were slower to escape, even before exposure to inescapable stress. They also spent more time immobile in the tail suspension test and less time in the lighted section of a light/dark box. The SCN-Bmal1-KD mice also showed greater weight gain, an abnormal circadian pattern of corticosterone, and an attenuated increase of corticosterone in response to stress. Disrupting SCN circadian rhythms is sufficient to cause helplessness, behavioral despair, and anxiety-like behavior in mice, establishing SCN-Bmal1-KD mice as a new animal model of depression.


Group A Streptococcus induces CD1a-autoreactive T cells and promotes psoriatic inflammation.
Group A Streptococcus (GAS) infection is associated with multiple clinical sequelae, including different subtypes of psoriasis. Such post-streptococcal disorders have been long known but are largely unexplained. CD1a is expressed at constitutively high levels by Langerhans cells and presents lipid antigens to T cells, but the potential relevance to GAS infection has not been studied. Here, we investigated whether GAS-responsive CD1a-restricted T cells contribute to the pathogenesis of psoriasis. Healthy individuals had high frequencies of circulating and cutaneous GAS-responsive CD4+ and CD8+ T cells with rapid effector functions, including the production of interleukin-22 (IL-22). Human skin and blood single-cell CITE-seq analyses of IL-22-producing T cells showed a type 17 signature with proliferative potential, whereas IFN-γ-producing T cells displayed cytotoxic T lymphocyte characteristics. Furthermore, individuals with psoriasis had significantly higher frequencies of circulating GAS-reactive T cells, enriched for markers of activation, cytolytic potential, and tissue association. In addition to responding to GAS, subsets of expanded GAS-reactive T cell clones/lines were found to be autoreactive, which included the recognition of the self-lipid antigen lysophosphatidylcholine. CD8+ T cell clones/lines produced cytolytic mediators and lysed infected CD1a-expressing cells. Furthermore, we established cutaneous models of GAS infection in a humanized CD1a transgenic mouse model and identified enhanced and prolonged local and systemic inflammation, with resolution through a psoriasis-like phenotype. Together, these findings link GAS infection to the CD1a pathway and show that GAS infection promotes the proliferation and activation of CD1a-autoreactive T cells, with relevance to post-streptococcal disease, including the pathogenesis and treatment of psoriasis.


Alcohol-metabolizing genes and alcohol phenotypes in an Israeli household sample.
Alcohol dehydrogenase 1B and 1C (ADH1B and ADH1C) variants have been robustly associated with alcohol phenotypes in East Asian populations, but less so in non-Asian populations where prevalence of the most protective ADH1B allele is low (generally <5%). Further, the joint effects of ADH1B and ADH1C on alcohol phenotypes have been unclear. Therefore, we tested the independent and joint effects of ADH1B and ADH1C on alcohol phenotypes in an Israeli sample, with higher prevalence of the most protective ADH1B allele than other non-Asian populations. A structured interview assessed lifetime drinking and alcohol use disorders (AUDs) in adult Israeli household residents. Four single nucleotide polymorphisms (SNPs) were genotyped: ADH1B (rs1229984, rs1229982, and rs1159918) and ADH1C (rs698). Regression analysis examined the association between alcohol phenotypes and each SNP (absence vs. presence of the protective allele) as well as rs698/rs1229984 diplotypes (also indicating absence or presence of protective alleles) in lifetime drinkers (n = 1,129). Lack of the ADH1B rs1229984 protective allele was significantly associated with consumption- and AUD-related phenotypes (OR = 1.77 for AUD; OR = 1.83 for risk drinking), while lack of the ADH1C rs698 protective allele was significantly associated with AUD-related phenotypes (OR = 2.32 for AUD). Diplotype analysis indicated that jointly ADH1B and ADH1C significantly influenced AUD-related phenotypes. For example, among those without protective alleles for ADH1B or ADH1C, OR for AUD was 1.87 as compared to those without the protective allele for ADH1B only and was 3.16 as compared to those with protective alleles for both ADH1B and ADH1C. This study adds support for the relationship of ADH1B and ADH1C and alcohol phenotypes in non-Asians. Further, these findings help clarify the mixed results from previous studies by showing that ADH1B and ADH1C jointly effect AUDs, but not consumption. Studies of the association between alcohol phenotypes and either ADH1B or ADH1C alone may employ an oversimplified model, masking relevant information.


Effects of certain antioxidants on lipid peroxidation process in lung homogenates of L thyroxine-receiving rats.
A possible role of antioxidants in thyreotoxicosis was investigated. We examined the parameters of lipid peroxidation (LPO): conjugated dienes (CD), malondialdehyde (MDA), Schiff bases (SB) in lung homogenates of male Wistar rats. Two control groups were created: Group 1 - intact animals and Group 2 - animals injected with 0,9% NaCl. In Experiment I, the animals received L-thyroxin (LT4) i.p. (Groups 3-7). After one week the rats received additionally: Group 4 - melatonin (MEL); Group 5 - propylthiouracil (PTU); Group 6 - Ambroxol (AMB); Group 7 - N-acetylocysteine (NAC). In Experiment II, the animals received only antioxidants. In Experiment I, we noticed a significantly higher MDA and SB level in Group 2, compared to that in Group 1. Moreover, we observed a significantly higher MDA and SB level in Group 3, vs. that in Group 1, but SB level was lower in Group 3 than in Group 2. Melatonin, PTU and NAC reduced CD; PTU, AMB diminished MDA and MEL, AMB lowered SB levels as compared to Group 3. In Experiment II, we observed significantly higher MDA and SB level in Group 2, vs. that in Group 1. Melatonin, AMB and NAC decreased MDA and SB level, when compared to Group 2 but PTU elevated MDA and SB level vs. that in Group 1. 1) L-T4 suppresses LPO, 2) MEL, AMB and NAC protect against LPO, 3) PTU is an antioxidant in thyreotoxicosis, however, when administered alone, it enhances LPO, 4) stress accelerate LPO.


Complementation between temperature-sensitive and deletion mutants of reovirus.
A very low level of complementation has been found in conventional crosses between various classes of temperature-sensitive (ts) mutants of reovirus. A more definitive test for complementation was devised through a plaque assay on cell monolayers mixedly infected with defective reovirions lacking the L1 segment and prototype ts mutants from one or other of the known classes of reovirus mutants. An increase in the number of plaques on the mixedly infected plates over that on control plates infected with defective virions or ts mutants alone indicated that the ts mutant had been complemented by the defective virus. Class A, B, D, F, and G mutants were complemented at 39 C by the defective viruses, whereas class C and E mutants were not. In tests to determine whether complementation was reciprocal it was found that the defective virions were complemented by a class G mutant but not by the class C mutant. This and previous work (D.A. Spandidos and A. F. Graham, 1975) has therefore shown that of the seven known classes of ts mutants the class C mutant is the only one that neither complements nor is complemented by the defective virions. For this reason the class C ts mutation has been assigned to the L1 segment of the viral genome.


Chromosomal instability confers intrinsic multidrug resistance.
Aneuploidy is associated with poor prognosis in solid tumors. Spontaneous chromosome missegregation events in aneuploid cells promote chromosomal instability (CIN) that may contribute to the acquisition of multidrug resistance in vitro and heighten risk for tumor relapse in animal models. Identification of distinct therapeutic agents that target tumor karyotypic complexity has important clinical implications. To identify distinct therapeutic approaches to specifically limit the growth of CIN tumors, we focused on a panel of colorectal cancer (CRC) cell lines, previously classified as either chromosomally unstable (CIN(+)) or diploid/near-diploid (CIN(-)), and treated them individually with a library of kinase inhibitors targeting components of signal transduction, cell cycle, and transmembrane receptor signaling pathways. CIN(+) cell lines displayed significant intrinsic multidrug resistance compared with CIN(-) cancer cell lines, and this seemed to be independent of somatic mutation status and proliferation rate. Confirming the association of CIN rather than ploidy status with multidrug resistance, tetraploid isogenic cells that had arisen from diploid cell lines displayed lower drug sensitivity than their diploid parental cells only with increasing chromosomal heterogeneity and isogenic cell line models of CIN(+) displayed multidrug resistance relative to their CIN(-) parental cancer cell line derivatives. In a meta-analysis of CRC outcome following cytotoxic treatment, CIN(+) predicted worse progression-free or disease-free survival relative to patients with CIN(-) disease. Our results suggest that stratifying tumor responses according to CIN status should be considered within the context of clinical trials to minimize the confounding effects of tumor CIN status on drug sensitivity.


Alzheimer's disease and memory strength: Gradual decline of memory traces as a function of their strength.
Episodic memory impairment is at the core of amnestic mild cognitive impairment (aMCI) and Alzheimer's disease (AD). The origin of memory deficits may result either from an encoding deficit or from an accelerated decline of the memory trace. The present study explores these two hypotheses. We used the delayed-matching-to sample 48 items (DMS-48) memory test in a group of controls, aMCI patients, and AD patients (n= 16 in each group). The DMS-48 offers an incidental learning phase followed by three forced-choice recognition tests at three-minute, one-hour, and one-week delays. Moreover, the forced-choice test distinguishes three kinds of couple items: abstract (meaningless), paired (two similar exemplars), and unique (two different objects) items. As predicted by the accelerated forgetting hypothesis, patients showed a decrease in recognition performance over time. Controls also exhibited a similar decline in performance. As predicted by the encoding deficit hypothesis, abstract items were the most poorly recognized in AD, at both the three-minute and the one-week delays. In AD, recognition of the paired items also dropped after the one-hour delay, followed by unique items after a one-week delay. Patients with aMCI exhibited a performance that was similar to controls, except for abstract items, which dropped at the one-week delay. These results are discussed in light of a third hypothesis, the memory strength hypothesis, in order to better account for the progressive decline in memory performance as a function of the item type in AD.


Modeling huddling penguins.
We present a systematic and quantitative model of huddling penguins. In this mathematical model, each individual penguin in the huddle seeks only to reduce its own heat loss. Consequently, penguins on the boundary of the huddle that are most exposed to the wind move downwind to more sheltered locations along the boundary. In contrast, penguins in the interior of the huddle neither have the space to move nor experience a significant heat loss, and they therefore remain stationary. Through these individual movements, the entire huddle experiences a robust cumulative effect that we identify, describe, and quantify. This mathematical model requires a calculation of the wind flowing around the huddle and of the resulting temperature distribution. Both of these must be recomputed each time an individual penguin moves since the huddle shape changes. Using our simulation results, we find that the key parameters affecting the huddle dynamics are the number of penguins in the huddle, the wind strength, and the amount of uncertainty in the movement of the penguins. Moreover, we find that the lone assumption of individual penguins minimizing their own heat loss results in all penguins having approximately equal access to the warmth of the huddle.


Histology image analysis for carcinoma detection and grading.
This paper presents an overview of the image analysis techniques in the domain of histopathology, specifically, for the objective of automated carcinoma detection and classification. As in other biomedical imaging areas such as radiology, many computer assisted diagnosis (CAD) systems have been implemented to aid histopathologists and clinicians in cancer diagnosis and research, which have been attempted to significantly reduce the labor and subjectivity of traditional manual intervention with histology images. The task of automated histology image analysis is usually not simple due to the unique characteristics of histology imaging, including the variability in image preparation techniques, clinical interpretation protocols, and the complex structures and very large size of the images themselves. In this paper we discuss those characteristics, provide relevant background information about slide preparation and interpretation, and review the application of digital image processing techniques to the field of histology image analysis. In particular, emphasis is given to state-of-the-art image segmentation methods for feature extraction and disease classification. Four major carcinomas of cervix, prostate, breast, and lung are selected to illustrate the functions and capabilities of existing CAD systems.


Gene expression profiling of subcutaneous adipose tissue in morbid obesity using a focused microarray: distinct expression of cell-cycle- and differentiation-related genes.
Obesity results from an imbalance between food intake and energy expenditure, which leads to an excess of adipose tissue. The excess of adipose tissue and adipocyte dysfunction associated with obesity are linked to the abnormal regulation of adipogenesis. The objective of this study was to analyze the expression profile of cell-cycle- and lipid-metabolism-related genes of adipose tissue in morbid obesity. We used a custom-made focused cDNA microarray to determine the adipose tissue mRNA expression profile. Gene expression of subcutaneous abdominal fat samples from 15 morbidly obese women was compared with subcutaneous fat samples from 10 nonobese control patients. The findings were validated in an independent population of 31 obese women and 9 obese men and in an animal model of obesity (Lepob/ob mice) by real-time RT-PCR. Microarray analysis revealed that transcription factors that regulate the first stages of adipocyte differentiation, such as CCAAT/enhancer binding protein beta (C/EBPβ) and JUN, were upregulated in the adipose tissues of morbidly obese patients. The expression of peroxisome proliferator-activated receptor gamma (PPARγ), a transcription factor which controls lipid metabolism and the final steps of preadipocyte conversion into mature adipocytes, was downregulated. The expression of three cyclin-dependent kinase inhibitors that regulate clonal expansion and postmitotic growth arrest during adipocyte differentiation was also altered in obese subjects: p18 and p27 were downregulated, and p21 was upregulated. Angiopoietin-like 4 (ANGPTL4), which regulates angiogenesis, lipid and glucose metabolism and it is know to increase dramatically in the early stages of adipocyte differentiation, was upregulated. The expression of C/EBPβ, p18, p21, JUN, and ANGPTL4 presented similar alterations in subcutaneous adipose tissue of Lepob/ob mice. Our microarray gene profiling study revealed that the expression of genes involved in adipogenesis is profoundly altered in the subcutaneous adipose tissue of morbidly obese subjects. This expression pattern is consistent with an immature adipocyte phenotype that could reflect the expansion of the adipose tissue during obesity.


Social, educational, and psychological correlates of weight status in adolescents.
The purpose of this research was to examine the social, educational, and psychological correlates of weight status in an adolescent population. It was hypothesized that obese adolescents would differ on psychological, social, and educational variables compared with their non-overweight peers. In this cross-sectional study, a population-based sample of 4742 male and 5201 female public school students in the 7th, 9th, and 11th grades responded anonymously to a classroom administered questionnaire. Body mass index was calculated from self-reported height and weight and categorized into four classes of weight status: underweight (<15th percentile), average weight (15th to 85th percentile), overweight (>85th to 95th percentile), and obese (>95th percentile). The questionnaire also included questions about social experiences, psychological well-being, educational experiences, and future goals. Associations of weight status with social, psychological, and educational variables and future goals were explored. After adjustment for grade level, race, and parental socioeconomic status, obese girls, when compared with their average weight counterparts, were 1.63 (95% confidence interval [CI]: 1.16, 2.30) times less likely to hang out with friends in the last week, 1.49 (95% CI: 1.12, 1.98) times more likely to report serious emotional problems in the last year, 1.79 (95% CI: 1.20, 2.65) times more likely to report hopelessness, and 1.73 (95% CI: 1.21, 1.98) times more likely to report a suicide attempt in the last year. Obese girls were also 1.51 (95% CI: 1.09, 2.10) times more likely to report being held back a grade and 2.09 (95% CI: 1.35, 3.24) times more likely to consider themselves poor students compared with average weight girls. Compared with their average weight counterparts, obese boys were 1.91 (95% CI: 1.43, 2.54) times less likely to hang out with friends in the last week, 1.34 (95% CI: 1.06, 1.70) times more likely to feel that their friends do not care about them, 1.38 (95% CI: 1.08, 1.76) times more likely to report having serious problems in the last year, 1.46 (95% CI: 1.05, 0.03) times more likely to consider themselves poor students, and 2.18 (95% CI: 1.45, 3.30) times more likely to expect to quit school. Compared with average weight boys, underweight boys were 1.67 (95% CI: 1.30, 2.13) times more likely to report hanging out with friends in the last week, 1.22 (95% CI: 1.01, 1.49) times more likely to report disliking school, and 1.40 (95% CI: 1.06, 1.86) times more likely to consider themselves poor students. Associations of weight status with social relationships, school experiences, psychological well-being, and some future aspirations were observed. Among girls, the pattern of observations indicates that obese girls reported more adverse social, educational, and psychological correlates. Obese as well as underweight boys also reported some adverse social and educational correlates. These findings contribute to an understanding of how adolescent experiences vary by weight status and suggest social and psychological risks associated with not meeting weight and body shape ideals embedded in the larger culture.


β-Cells are not uniform after all-Novel insights into molecular heterogeneity of insulin-secreting cells.
While the β-cells of the endocrine pancreas are defined as cells with high levels of insulin production and tight stimulus-secretion coupling, the existence of functional heterogeneity among them has been known for decades. Recent advances in molecular technologies, in particular single-cell profiling on both the protein and messenger RNA level, have uncovered that β-cells exist in several antigenically and molecularly definable states. Using antibodies to cell surface markers or multidimensional clustering of β-cells using more than 20 protein markers by mass cytometry, 4 distinct groups of β-cells could be differentiated. However, whether these states represent permanent cell lineages or are readily interconvertible from one group to another remains to be determined. Nevertheless, future analysis of the pathogenesis of type 1 and type 2 diabetes will certainly benefit from a growing appreciation of β-cell heterogeneity. Here, we aim to summarize concisely the recent advances in the field and their possible impact on our understanding of β-cell physiology and pathophysiology.


Retrieval of aerosol profile variations from reflected radiation in the oxygen absorption a band.
Through a Fredholm integral equation of the first kind, aerosol kernel functions relate the variations in radiance measured by satellites to the variations in the aerosol extinction profile and thus permit profile retrieval from radiance measurements by inversion of the set of radiative transfer equations for various spectral intervals. Previously [Appl. Opt. 36, 1328 (1997)] the kernel functions were evaluated for the red and near-infrared spectral regions outside molecular absorption bands. Here they are computed within the oxygen A band with 20-cm(-1) spectral resolution. It is shown that, even with such a relatively low spectral resolution, the new set of kernels is able to provide better information on and improved accuracy of the retrieved profile.


Inflammatory markers and exposure to airborne particles among workers in a Swedish pulp and paper mill.
To study the relationship between exposure to airborne particles in a pulp and paper mill and markers of inflammation and coagulation in blood. Personal sampling of inhalable dust was performed for 72 subjects working in a Swedish pulp and paper mill. Stationary measurements were used to study concentrations of total dust, respirable dust, PM10 and PM2.5, the particle surface area and the particle number concentrations. Markers of inflammation, interleukins (IL-1b, IL-6, IL-8, and IL-10), C-reactive protein (CRP), serum amyloid A (SAA), and fibrinogen and markers of coagulation factor VIII, von Willebrand, plasminogen activator inhibitor, and D-dimer were measured in plasma or serum. Sampling was performed on the last day of the work free period of 5 days, before and after the shift the first day of work and after the shifts the second and third day. In a mixed model analysis, the relationship between particulate exposures and inflammatory markers was determined. Sex, age, smoking, and BMI were included as covariates. The average 8-h time-weighted average (TWA) air concentration levels of inhalable dust were 0.30 mg/m(3), range 0.005-3.3 mg/m(3). The proxies for average 8-h TWAs of respirable dust were 0.045 mg/m(3). Significant and consistent positive relations were found between several exposure metrics (PM 10, total and inhalable dust) and CRP, SAA and fibrinogen taken post-shift, suggesting a dose-effect relationship. This study supports a relationship between occupational particle exposure and established inflammatory markers, which may indicate an increased risk of cardiovascular disease.


Increased platelet aggregation and turnover in the acute phase of ST-elevation myocardial infarction.
Newly produced platelets are present in the acute phase of ST-elevation myocardial infarction (STEMI). This may influence the antiplatelet effect of aspirin and clopidogrel administered prior to primary percutaneous coronary intervention (PPCI). The aims of this study were to investigate the antiplatelet effect of aspirin and clopidogrel and evaluate platelet turnover in the acute phase of STEMI compared to a stable phase 3 months later. In this observational follow-up study on 48 STEMI patients transferred for PPCI, loading doses of aspirin (300 mg) and clopidogrel (600 mg) were given orally in the ambulance. Blood samples were obtained immediately prior to PPCI, at 4 and 12 hours after administration of bolus doses and at follow-up after 3 months. Residual platelet aggregation was evaluated by Multiplate® and VerifyNow® aggregometry. Platelet turnover was evaluated by automated flow cytometry. In the acute phase of STEMI at the time of PPCI, residual platelet aggregation was significantly higher prior to PPCI compared to 4 and 12 hours after loading doses of aspirin and clopidogrel and 3 months later (p-values < 0.01). Furthermore, platelet turnover indices (mean platelet volume, immature platelet fraction, and immature platelet count) were increased in the acute phase of STEMI compared to 3 months after PPCI (p-values < 0.0001). In conclusion, residual platelet aggregation was high in the acute phase of STEMI despite dual antiplatelet treatment with aspirin and clopidogrel. Increased platelet turnover may partly explain the reduced efficacy of antiplatelet drugs in the acute phase of STEMI.