Isolation and characterization of copolymers of beta-lactoglobulin, alpha-lactalbumin, kappa-casein, and alphas1-casein generated by pressurization and thermal treatment of raw milk.
Raw skim milk was submitted to high pressure (300 to 600 MPa) and temperature (4 to 70 degrees C) treatments for 2 or 5 min. The combined effects of pressure and temperature on milk proteins induced structural changes and polymer and copolymer formation characterized by anion-exchange and size-exclusion fast protein liquid chromatography and electrophoretic techniques. Approximately half of the beta-lactoglobulin formed polymers, and the other half formed large copolymers, mainly with kappa-casein, alpha-lactalbumin via intermolecular disulfide bond exchange, and alpha(s1)-casein via physicochemical interactions, in proportions of 1.0:0.7:0.3:0.1, respectively. Minor whey proteins (serum albumin, immunoglobulins, and lactoferrin) also participated in the formation of the copolymers but to a lesser extent. Two populations of the copolymers were found with apparent molecular masses ranging from 440 to 2000 kDa for the first and more than 2000 kDa for the second. On the contrary, for heated milks the aggregation kinetics obtained by combination of high pressure and thermal treatment were very fast, as no intermediates such as dimers and small size oligomers were observed after pressurization, whatever the temperature studied. Lactosylation of proteins as well as proteolysis were very limited. A beta-casein amino-terminal peptide of 22 kDa was specifically recovered in milk samples treated under the more drastic conditions (500 MPa/55 degrees C per 5 min and 600 MPa/70 degrees C per 5 min) and might have been generated by neutral proteases such as elastase released from somatic cells present in milk. No casein was released from the micelle whatever the combination of high pressure and temperature studied.


Smart textiles in wound care: functionalization of cotton/PET blends with antimicrobial nanocapsules.
Management of infected wounds is one of the most costly procedures in the health care sector. Burn wounds are of significant importance due to the high infection risk that can possibly lead to severe consequences such as sepsis. Because antibiotic wound treatments have caused increasing antibiotic resistance in bacteria, there is currently a strong need for alternative strategies. Therefore, we developed new antimicrobial wound dressings consisting of pH-responsive human serum albumin/silk fibroin nanocapsules immobilized onto cotton/polyethylene terephthalate (PET) blends loaded with eugenol, which is an antimicrobial phenylpropanoid. Ultrasound-assisted production of eugenol-loaded nanocapsules resulted in particle sizes (hydrodynamic radii) between 319.73 ± 17.50 and 574.00 ± 92.76 nm and zeta potentials ranging from -10.39 ± 1.99 mV to -12.11 ± 0.59 mV. Because recent discoveries have indicated that the sweat glands contribute to wound reepithelialisation, release studies of eugenol were conducted in different artificial sweat formulas that varied in pH. Formulations containing 10% silk fibroin with lower degradation degree exhibited the highest release of 41% at pH 6.0. After immobilization, the functionalized cotton/PET blends were able to inhibit 81% of Staphylococcus aureus and 33% of Escherichia coli growth. Particle uniformity, silk fibroin concentration, and high surface-area-to-volume ratio of the produced nanocapsules were identified as the contributing factors leading to high antimicrobial activities against both strains. Therefore, the production of antimicrobial textiles using nanocapsules loaded with an active natural compound that will not contribute to antibiotic resistance is seen as a potential future alternative to commercially available antiseptic wound dressings.


Quantitative trait loci analysis for plasma HDL-cholesterol concentrations and atherosclerosis susceptibility between inbred mouse strains C57BL/6J and 129S1/SvImJ.
The C57BL/6 (B6) and 129 mouse inbred strains differ markedly in plasma HDL-cholesterol concentrations and atherosclerosis susceptibility after a high-fat diet consumption. To identify loci controlling these traits, we performed quantitative trait loci (QTL) analysis. We fed a high-fat diet to 294 (B6x129S1/SvImJ)F2 females for 14 weeks, measured plasma HDL concentrations and size of aortic fatty-streak lesions, genotyped F2 females, and performed QTL analysis. HDL concentrations were affected by six loci: Hdlq14 and Hdlq15 on chromosome 1 (peaks cM 80 and cM 104, logarithm of odds [LOD] 5.3 and 9.7, respectively); Hdlq16 on chromosome 8 (cM 44, LOD 2.6); Hdlq17 on chromosome 9 (cM 24, LOD 2.9); Hdlq18 on chromosome 12 (cM 20, LOD 5.9); and Hdlq19 on chromosome 2 (cM 90), which interacted with Hdlq15. Atherosclerosis susceptibility was affected by five loci: Ath17 on chromosome 10 (cM 34, LOD 6.6); Ath18 on chromosome 12 (cM 16, LOD 3.7); Ath19 (chromosome 11, cM 60), which interacted with Ath18; and Ath20 (chromosome 10, cM 10), which interacted with Ath21 (chromosome 12, cM 50). We identified six loci for HDL and five loci for atherosclerosis susceptibility in a (B6x129S1/SvImJ)F2 intercross.


Characterization and Evaluation of Skin Permeation of Tocopheryl Phosphoric Acid-loaded Phospholipid Nanoparticles.
Tocopheryl phosphoric acid (TPA, a hydrophilic vitamin E derivative) loaded liposome and glycerin containing phospholipid nanoparticles (GPLNP) were prepared using the film rehydration and extrusion method. Nanoparticle formulations were evaluated for size, zeta potential, and in vitro permeation across hairless mouse skin, and 31P NMR spectral analysis was performed. The prepared formulations were stable for 2 weeks, and their mean nanoparticle size varied between 90 and 140 nm. Although glycerin did not affect the particle size of the empty (no TPA) system, TPA-loading resulted in the reduction of particle size and conferred a negative charge. The 31P NMR spectral analysis showed that the presence of glycerin in the formulation changed the nanoparticle structure from a bilayer to a nonbilayer. Moreover, it was suggested that TPA molecules interacted with phospholipid by entrapping nanoparticles in the formulations. TPA did not permeate across the hairless mouse skin after 48 h. However, the TPA concentration in the hairless mouse skin after permeation study increased in the nanoparticle systems and the 30% GPLNP formulation was the best formulation for the accelerated TPA permeation in the hairless mouse skin. These results demonstrate that 30% GPLNP improved TPA permeation in the hairless mouse skin model. And it was strongly suggested that glycerin has an important role for changing the structure of nanoparticles and enhancing the skin permeation of TPA.


Arterial baroreceptor reflexes in young and old racing greyhounds.
The characteristics of the carotid sinus baroreceptor (CSB) reflex control of arterial pressure-flow relations were studied in young (18 mo) and old (10.5 yr) racing greyhounds anesthetized with chloralose. Pressure and flow were simultaneously measured in the ascending aorta, the celiac, superior mesenteric, renal, and iliac arteries under steady-state conditions of pulsatile perfusion of the isolated carotid sinuses before and after bilateral cervical vagotomy. Operating-point (OP) values of mean pressure were not significantly different between the young or old greyhounds, but OP values of cardiac output were lower and of peripheral resistance were higher in the older animals. OP values of regional resistances were higher in the older animals, but only those of the celiac and mesenteric beds were significantly different. Values of OP sensitivity of central hemodynamic variables were not decreased in the older animals. OP values of renal and iliac resistance sensitivity tended to be lower in the older animals. The overall range of control of the various hemodynamic variables by the CSB was lower in the older group only in the case of iliac resistance. The characteristics of the CSB were not uniformly depressed in the older animals but were relatively well maintained compared with those of the younger group.


The effect of olfactory stimuli on the balance ability of stroke patients.
[Purpose] The present study attempted to identify the effect of olfactory stimulation on the balance ability of stroke patients. [Subjects] Thirty-three (33 males) stroke patients participated in the study. The stroke patients were divided into three groups: a black pepper oil (BPO) group (n=11), lavender oil (LVO) group (n=11), and distilled water (DW) group (n=11). [Methods] Two sessions (control trial/stimulus trial) of Romberg's test (eyes open 1 min/eyes closed 1 min) were conducted on a force platform to measure the data for the COP (center of pressure). Olfactory stimulation was provided at as a stimulus. [Results] With the eyes open, a statistically significant difference was found in average anterior posterior displacement (Ymean) and average medial lateral displacement (Xmean) among the three groups when comparing the groups before and after stimulation. The comparison between the eyes open and eyes closed conditions in each group showed a significant difference in the area of the 95% confidence ellipse (area) and Xmean of the BPO group and in the area of the LVO group (area, Xmean). [Conclusion] The findings indicate that the interaction of brain areas activated by the olfactory stimulation exerts an influence on the balance ability of stroke patients.


Laterality of expression in portraiture: putting your best cheek forward.
Portraits, both photographic and painted, are often produced with more of one side of the face showing than the other. Typically, the left side of the face is overrepresented, with the head turned slightly to the sitter's right. This leftward bias is weaker for painted male portraits and non-existent for portraits of scientists from the Royal Society. What mechanism might account for this bias? Examination of portraits painted by left- and right-handers and of self-portraits suggests that the bias is not determined by a mechanical preference of the artist or by the viewer's aesthetics. The leftward bias seems to be determined by the sitters and their desire to display the left side of their face, which is controlled by the emotive, right cerebral hemisphere. When we asked people to portray as much emotion as possible when posing for a family portrait, they tended to present the left side of their face. When asked to pose as scientists and avoid portraying emotion, participants tended to present their right side. The motivation to portray emotion, or conceal it, might explain why portraits of males show a reduced leftward bias, and also why portraits of scientists from the Royal Society show no leftward bias.


Pediatric emergency department utilization within a statewide medicaid managed care system.
To describe pediatric emergency department (ED) utilization within a Medicaid managed care system and to investigate the association between pediatric ED utilization and the type of medical home. This was a retrospective cohort study, set at Rhode Island's statewide Medicaid managed care program (RIte Care). The study population was a random sample of 2,000 children aged 19 to 35 months at the conclusion of a period of continuous RIte Care enrollment from July 1, 1996, until June 30, 1997. For description of ED utilization, rates of utilization and percentages were calculated. Bivariate and multivariate analyses were used to determine the effect of type of medical home and individual/family characteristics on ED utilization. Logistic regression models were used to determine odds ratios for types of medical home while controlling for sociodemographic factors and practice level variation. Of the 1,988 children who were eligible for analysis, 791 children utilized the ED during the study year at a rate of 68 ED visits/100 person-years. Thirty-one percent of those visits were for injuries/poisonings. Logistic regression analysis demonstrated decreased ED utilization for patients enrolled in a staff-model health maintenance organization (HMO) (OR 0.34; 95% CI = 0.27 to 0.42) or a health center (OR 0.71; 95% CI = 0.53 to 0.95) compared with hospital-based clinics. There was no significant association between the receipt of preventive services and ED utilization. Despite the presence of a defined medical home, ED use was common among preschool children enrolled in the statewide Medicaid managed care program. The type of medical home was an important determinant of ED use in this population.


Renal osteodystrophy in the first decade of the new millennium: analysis of 630 bone biopsies in black and white patients.
Renal osteodystrophy occurs early during loss of kidney function. There are 26 million American patients with chronic kidney disease (CKD), and almost all patients with CKD stage 5 have abnormal bone histology. Six hundred and thirty bone biopsies from adult CKD-5 patients on dialysis were evaluated by histomorphometry and analyzed using the turnover (T), mineralization (M), and volume (V) classification. There were racial differences; whites exhibited predominantly low turnover (62%), whereas blacks showed mostly normal or high turnover (68%). A mineralization defect was observed in only 3% of patients. In whites, cancellous bone volume was low, normal, or high in approximately the same number of patients, whereas in blacks, cancellous bone volume was high in two-thirds of the patients. More than 80% of blacks and whites with low cancellous bone volume had thin trabeculae owing to low bone formation. Cortical thickness was low in half the whites, whereas it was normal in three-quarters of blacks. Cortical porosity was high in 50% of whites, whereas three-quarters of blacks had high porosity. In summary, the TMV system gives relevant information. It should be expanded to include the architecture of cancellous and cortical bone. There are racial differences. Low bone volume and low bone turnover are more frequent than heretofore appreciated, whereas mineralization defects nowadays are observed rarely in adults. These findings call for an adjustment of the current therapeutic paradigm that takes into consideration race and risk of low bone volume and turnover. The latter have been shown to be associated with increased vascular calcifications.


Pathomechanisms and management of osteoporotic pain with no traumatic evidence.
Osteoporosis is a pathological state with an unbalanced bone metabolism mainly caused by accelerated osteoporotic osteoclast activity due to a postmenopausal estrogen deficiency, and it causes some kinds of pain, which can be divided into two types: traumatic pain due to a fragility fracture from impaired rigidity, and pain derived from an osteoporotic pathology without evidence of fracture. We aimed to review the concepts of osteoporosis-related pain and its management. We reviewed clinical and basic articles on osteoporosis-related pain, especially with a focus on the mechanism of pain derived from an osteoporotic pathology (i.e., osteoporotic pain) and its pharmacological treatment. Osteoporosis-related pain tends to be robust and acute if it is due to fracture or collapse, whereas pathology-related osteoporotic pain is vague and dull. Non-traumatic osteoporotic pain can originate from an undetectable microfracture or structural change such as muscle fatigue in kyphotic patients. Furthermore, basic studies have shown that the osteoporotic state itself is related to pain or hyperalgesia with increased pain-related neuropeptide expression or acid-sensing channels in the local tissue and nervous system. Traditional treatment for osteoporotic pain potentially prevents possible fracture-induced pain by increasing bone mineral density and affecting related mediators such as osteoclasts and osteoblasts. The most common agent for osteoporotic pain management is a bisphosphonate. Other non-osteoporotic analgesic agents such as celecoxib have also been reported to have a suppressive effect on osteoporotic pain. Osteoporotic pain has traumatic and non-traumatic factors. Anti-osteoporotic treatments are effective for osteoporotic pain, as they improve bone structure and the condition of the pain-related sensory nervous system. Physicians should always consider these matters when choosing a treatment strategy that would best benefit patients with osteoporotic pain.


Predicting outcomes following cognitive behaviour therapy in child anxiety disorders: the influence of genetic, demographic and clinical information.
Within a therapeutic gene by environment (G × E) framework, we recently demonstrated that variation in the Serotonin Transporter Promoter Polymorphism; 5HTTLPR and marker rs6330 in Nerve Growth Factor gene; NGF is associated with poorer outcomes following cognitive behaviour therapy (CBT) for child anxiety disorders. The aim of this study was to explore one potential means of extending the translational reach of G × E data in a way that may be clinically informative. We describe a 'risk-index' approach combining genetic, demographic and clinical data and test its ability to predict diagnostic outcome following CBT in anxious children. DNA and clinical data were collected from 384 children with a primary anxiety disorder undergoing CBT. We tested our risk model in five cross-validation training sets. In predicting treatment outcome, six variables had a minimum mean beta value of 0.5:5HTTLPR, NGF rs6330, gender, primary anxiety severity, comorbid mood disorder and comorbid externalising disorder. A risk index (range 0-8) constructed from these variables had moderate a predictive ability (AUC = .62-.69) in this study. Children scoring high on this index (5-8) were approximately three times as likely to retain their primary anxiety disorder at follow-up as compared with those children scoring 2 or less. Significant genetic, demographic and clinical predictors of outcome following CBT for anxiety-disordered children were identified. Combining these predictors within a risk index could be used to identify which children are less likely to be diagnosis-free following CBT alone and require longer or enhanced treatment. The 'risk-index' approach represents one means of harnessing the translational potential of G × E data.


Leading the Way in Exercise and Diet (Project LEAD): intervening to improve function among older breast and prostate cancer survivors.
The U.S. population is aging, bringing with it an increased prevalence of chronic disease and concomitant declines in physical function. The risk of developing cancer increases significantly with age, and functional decline is much more likely once a cancer diagnosis is rendered. Thus, functional status in later life is a key concern, one that is heightened among elders who have been diagnosed with cancer. To date, however, there have been few trials that have exclusively addressed issues related to cancer survivorship among older cancer patients, and to our knowledge, none has focused on preserving or enhancing physical functioning. This paper describes the study design and methodological considerations of a randomized controlled trial to determine if a personally tailored workbook and telephone counseling program can positively affect physical activity and dietary behaviors and ultimately the physical functioning of up to 420 older men and women newly diagnosed with breast or prostate cancer. This trial is unique because the cancer diagnosis is used not only as a marker of risk for functional decline, but also as a "teachable moment" - an opportune time when elders may be more receptive to making beneficial lifestyle changes. Undoubtedly, as cure rates for cancer increase and intersect with ever-growing numbers of elderly, there will be numerous opportunities to provide and test interventions within this vulnerable population and to target functional status as a primary outcome. In reporting our methods, we hope to give others "a leg up," so that they can hurdle with greater ease the barriers we experienced, and thus advance the field more rapidly.


Age specific prediction of maximal oxygen uptake in boys.
Maximal oxygen uptake was determined in 62 boys each year from age 11 to 15 years. Multiple regression analyses were used to predict treadmill VO2 max at yearly age intervals from size measures and submaximal cycle heart rates. Derived equations were validated in an independent sample of 98 boys of the same age range. In boys aged 12 to 15 years approximately 80% or more of the variance in VO2 max was accounted for by age specific equations employing mass, height and skinfolds as explanatory variables. Prediction of VO2max was not as strong for 10 and 11 year olds, with less than 70% of the variance explained. Measures of VO2 or work rate and heart rate were not useful in predicting VO2max either by themselves or in conjunction with the size measures. The findings clearly demonstrated that submaximal exercise testing to predict VO2max, or cardiorespiratory fitness, in boys was of questionable value. The prediction of VO2max in boys aged 12 to 15 by in the independent sample was shown to have a mean absolute error of 6 to 8% while the value for age 11 y was 10%. The error is similar in magnitude to that of predicting VO2max with an exercise test in adults. The present age specific equations resulted in smaller errors of estimation than found in previous studies of VO2max estimation in children.


Clinical and biochemical characterization of women with polycystic ovary syndrome in North Rhine-Westphalia.
Polycystic ovary syndrome (PCOS), defined as the combination of oligoanovulation and hyperandrogenism, affects more than 5% of women of reproductive age. Insulin resistance and hyperinsulinemia appear to play an important role in its pathogenesis. Here, we will present a characterization of a PCOS cohort from North Rhine-Westphalia in Germany. Clinical features, family history as well as endocrine and metabolic parameters were prospectively recorded from 200 successive patients. All patients were evaluated for insulin resistance and beta-cell-function by oral glucose tolerance test. Patient data were compared with those of 98 age-matched control women. PCOS patients showed significantly higher BMI, body fat mass and androgen levels as well as impaired glucose and insulin metabolism. A positive family history of PCOS and diabetes was more frequent in PCOS patients. Insulin resistance (71%) was the most common metabolic abnormality in PCOS patients followed by obesity (52%) and dyslipidemia (46.3%), with an incidence of 31.5% for the metabolic syndrome. C-reactive protein and other cardiovascular risk factors were frequently elevated even in young PCOS patients. While the clinical characteristics and endocrine parameters of this German PCOS cohort were heterogeneous, they were comparable to those from other Caucasian populations.


Iron deficiency and child development.
Iron deficiency is widespread in infants and young children, especially in developing countries. Animal models provide convincing evidence that, despite iron repletion, iron deficiency during the brain growth spurt alters metabolism and neurotransmission, myelination, and gene and protein profiles. In the human, there is compelling evidence that 6- to 24-month-old infants with iron-deficiency anemia are at risk for poorer cognitive, motor, social-emotional, and neurophysiologic development in the short- and long-term outcome. In contrast to inconsistent developmental effects of iron therapy for iron-deficient infants, recent large, randomized trials of iron supplementation in developing countries uniformly show benefits of iron, especially on motor development and social-emotional behavior. These results indicate that adverse effects can be prevented and/or reversed with iron earlier in development or before iron deficiency becomes severe or chronic. New findings also point to the need for more attention to the developmental effects of prenatal iron deficiency.


Predicting bull fertility using genomic data and biological information.
The genomic prediction of unobserved genetic values or future phenotypes for complex traits has revolutionized agriculture and human medicine. Fertility traits are undoubtedly complex traits of great economic importance to the dairy industry. Although genomic prediction for improved cow fertility has received much attention, bull fertility largely has been ignored. The first aim of this study was to investigate the feasibility of genomic prediction of sire conception rate (SCR) in US Holstein dairy cattle. Standard genomic prediction often ignores any available information about functional features of the genome, although it is believed that such information can yield more accurate and more persistent predictions. Hence, the second objective was to incorporate prior biological information into predictive models and evaluate their performance. The analyses included the use of kernel-based models fitting either all single nucleotide polymorphisms (SNP; 55K) or only markers with presumed functional roles, such as SNP linked to Gene Ontology or Medical Subject Heading terms related to male fertility, or SNP significantly associated with SCR. Both single- and multikernel models were evaluated using linear and Gaussian kernels. Predictive ability was evaluated in 5-fold cross-validation. The entire set of SNP exhibited predictive correlations around 0.35. Neither Gene Ontology nor Medical Subject Heading gene sets achieved predictive abilities higher than their counterparts using random sets of SNP. Notably, kernel models fitting significant SNP achieved the best performance with increases in accuracy up to 5% compared with the standard whole-genome approach. Models fitting Gaussian kernels outperformed their counterparts fitting linear kernels irrespective of the set of SNP. Overall, our findings suggest that genomic prediction of bull fertility is feasible in dairy cattle. This provides potential for accurate genome-guided decisions, such as early culling of bull calves with low SCR predictions. In addition, exploiting nonlinear effects through the use of Gaussian kernels together with the incorporation of relevant markers seems to be a promising alternative to the standard approach. The inclusion of gene set results into prediction models deserves further research.


Diclofenac vs oxybuprocaine eyedrops for analgesia in paediatric strabismus surgery.
Forty children undergoing strabismus surgery as day patients were randomly allocated to receive oxybuprocaine 0.4% eyedrops or 0.1% diclofenac eyedrops for perioperative analgesia. A non-invasive anaesthetic technique using the reinforced laryngeal mask airway was used. The study demonstrated that both topical analgesics provided good to excellent analgesia and the anaesthetic technique was associated with a relatively low incidence of nausea and vomiting. Complications were limited to two children who were admitted with persistent postoperative nausea and vomiting.


Effect of tunicamycin on production by mouse fibroblast L929 cells of the factor-stimulating differentiation of mouse myeloid leukemic cells and the colony-stimulating factor.
Mouse myeloid leukemic M1 cells can be induced to differentiate into macrophages and granulocytes in vitro by a factor(s) stimulating differentiation of the cells (D-factor), which is suggested to be a glycoprotein. On the other hand, growth and differentiation of normal precursor cells of macrophages and granulocytes can be stimulated by a glycoprotein termed colony-stimulating factor (CSF). Mouse fibroblast L929 cells were found to produce both the D-factor and CSF. The properties of the D-factor and CSF and the roles of carbohydrates in the molecules of these factors were examined using tunicamycin, a specific inhibitor of asparaginase-linked glycosylation. Although both the D-factor and CSF were produced by L-cells in usual medium containing fetal calf serum, production of D-factor, but not CSF, was reduced by omission of serum from the medium. The activity of the D-factor was slightly decreased by treating the L-cells with tunicamycin (0.5 microgram/ml) in the presence of 2% fetal calf serum, without any decrease in CSF activity. Conditioned medium of L-cells incubated with or without tunicamycin was fractionated by gel filtration on a Sephadex G-200 column. Normal D-factor appeared as a single peak with an apparent molecular weight of 67,000. D-factor produced in the presence of tunicamycin had an apparent molecular weight of 25,000. On the other hand, most of the CSF was eluted in the void volume, even when it was produced in the presence of tunicamycin. The D-factor produced in the presence of tunicamycin was more sensitive than normal D-factor was to trypsin or heat treatment at 70 degrees. The CSF produced in the presence of tunicamycin was resistant to these treatments. These results indicate that the D-factor is distinct from CSF. Furthermore, the results suggest that the D-factor produced by L-cells is also a glycoprotein and that, although carbohydrate is not essential for production or activity of the D-factor, it contributes to stabilizing the protein portion of D-factor.


Rapid detection and identification of Staphylococcus aureus from blood culture specimens using real-time fluorescence PCR.
Molecular surveillance of pathogens has shown the need for rapid and dependable methods for the detection and identification of organisms of clinical and epidemiologic importance. Staphylococcus aureus, one of the most frequent causes of human infections, was used as a model organism to develop and refine a real-time fluorescence PCR assay and enhanced DNA purification method. One hundred clinical isolates of S. aureus, verified by biochemical reactions and latex agglutination and 90 negative control clinical isolates were screened in the assay. Moreover, fifty blood broth samples from blood culture bottles showing Gram-positive cocci in clusters on direct Gram's stain and 25 showing Gram-negative bacilli were screened. The probes, constructed from the nuc gene, correctly detected all S. aureus genomes present without cross-reaction to negative controls. The speed and ease of this approach will make it adaptable to identification of many bacterial pathogens and provide potential for adaptation to direct detection from other types of clinical specimens.


Climate change and temperature-dependent sex determination: can individual plasticity in nesting phenology prevent extreme sex ratios?
Under temperature-dependent sex determination (TSD), temperatures experienced by embryos during development determine the sex of the offspring. Consequently, populations of organisms with TSD have the potential to be strongly impacted by climatic warming that could bias offspring sex ratio, a fundamental demographic parameter involved in population dynamics. Moreover, many taxa with TSD are imperiled, so research on this phenomenon, particularly long-term field study, has assumed great urgency. Recently, turtles with TSD have joined the diverse list of taxa that have demonstrated population-level changes in breeding phenology in response to recent climate change. This raises the possibility that any adverse impacts of climate change on populations may be alleviated by individual plasticity in nesting phenology. Here, we examine data from a long-term study on a population of painted turtles (Chrysemys picta) to determine whether changes in phenology are due to individual plasticity and whether individual plasticity in the timing of nesting has the capacity to offset the sex ratio effects of a rise in climatic temperature. We find that individual females show plasticity in the date of first nesting each year, and that this plasticity depends on the climate from the previous winter. First nesting date is not repeatable within individuals, suggesting that it would not respond to selection. Sex ratios of hatchlings within a nest declined nonsignificantly over the nesting season. However, small increases in summer temperature had a much stronger effect on nest sex ratios than did laying nests earlier in the season. For this and other reasons, it seems unlikely that individual plasticity in the timing of nesting will offset the effects of climate change on sex ratios in this population, and we hypothesize that this conclusion applies to other populations with TSD.


Prevalence of glucose metabolism abnormalities and cardiovascular co-morbidity in the US elderly adult population.
To estimate the prevalence of glucose metabolism abnormalities, including diabetes, and its associated cardiovascular risk factors and co-morbidity in the US elderly population. Cross-sectional analysis of data from the Third National Health and Nutrition Examination Survey (NHANES III, 1988-1994) in adults aged 65 years and more. The 1997 American Diabetes Association (ADA) and the 1998 World Health Organization (WHO) criteria were used to classify the subject's glucose metabolism status. The 2-hour oral glucose tolerance test (OGTT) was performed only among those participants aged 40 to 74 years. The age-adjusted prevalence of diagnosed diabetes was 12.5% (95% CI: 11.4%-13.6%) among US adults aged 65 years or more. According to the ADA definition 40% of men and 28% of women were affected by some degree of glucose metabolism impairment. According to the WHO definition, 55% of men and 50% of women aged 65 to 74 years were affected by glucose metabolism abnormalities. Mexican-Americans were the most affected under both definitions (51% and 77%, respectively). Overall, 72% of elderly diagnosed diabetics had hypertension, 28% had coronary heart disease (CHD), 47% suffered from cardiovascular disease and 80% of them presented known CHD or multiple coronary risk factors, other than age, level of LDL-cholesterol and diabetes. Under both definitions, a trend towards a worsening coronary risk profile with increased glucose metabolism impairment was observed. A notable proportion of elderly people is affected by some degree of glucose metabolism impairment which in turn is associated with cardiovascular co-morbidity.


The effect of gentamicin on cytoskeletons in the vestibular sensory cells: a high-resolution scanning electron microscopic investigation.
The effect of gentamicin on the cytoskeletal organization of guniea pig vestibular sensory cells was investigated employing the saponin perfusion method and scanning electron microscopy. The intermediate filaments and microtubules were noted to have degenerated following gentamicin intoxication. Such degeneration was not linked with primary mitochondrial damage, but closely related to subsequent degeneration of Golgi apparatus and of endoplasmic reticulum. These findings suggest that microtubules and intermediate filaments work closely together to maintain the structural integrity of both the Golgi apparatus and the membrane-bound organelles, which can be altered by ototoxic drugs before the degeneration of membrane-bound organelles.


Co-metabolism of fluorobenzoates by natural microbial populations.
Co-metabolic degradation of monofluorobenzoates was carried out by a mixed soil population in a basal salts medium. The monofluorobenzoates did not support growth of microorganisms but were shown to be subject to ring cleavage as a result of microbial activity. Rate of ring cleavage was increased by use of the co-substrate enrichment technique using glucose as the co-substrate. Results indicate that the monofluorobenzoates were subject to an initial co-metabolic attack with glucose, providing the energy necessary for co-metabolism to proceed to a point where complete metabolism became possible.


Osteogenic effect of low-intensity pulsed ultrasound and whole-body vibration on peri-implant bone. An experimental in vivo study.
The aims of this study were (i) to compare the osteogenic impact of low-intensity pulsed ultrasound (LIPUS) and low-magnitude high-frequency (LMHF) loading achieved with whole-body vibration (WBV) on peri-implant bone healing and implant osseointegration in rat tibiae, and (ii) to examine their combined effect on these processes. Titanium implants were inserted in the bilateral tibiae of 28 Wistar rats. Rats were randomly divided into four groups: LIPUS + WBV, LIPUS, WBV, and control. LIPUS was applied to the implant placement site for 20 min/day on 5 days/week (1.5 MHz and 30 mW/cm2 ). WBV was applied for 15 min/day on 5 days/week (50 Hz and 0.5 g). In the LIPUS + WBV group, both stimuli were applied under the same stimulation conditions as in the LIPUS and WBV groups. After 4 weeks of treatment, peri-implant bone healing and implant osseointegration were assessed using removal torque (RT) tests, micro-CT analyses of relative gray (RG) value, and histomorphometrical analyses of bone-to-implant contact (BIC) and peri-implant bone formation (BV/TV). The LIPUS + WBV group had significantly greater BIC than the WBV and control groups. Although there were no significant intergroup differences in RT, RG value, and BV/TV, these variables tended to be greater in the LIPUS + WBV group than the other groups. The combination of LIPUS and LMHF loading may promote osteogenic activity around the implant. However, further study of the stimulation conditions of LIPUS and LMHF loading is necessary to better understand the osteogenic effects and the relationship between the two stimuli.


Basic adsorption heat exchanger theory for performance prediction of adsorption heat pumps.
Adsorption modules are the core components of thermally driven adsorption heat pumps and chillers. Due to the transient nature of the adsorption and desorption processes, usually complicated numerical models are used for prediction of efficiency and heat flow rates. In this research article, we suggest a radically simplified calculation based on splitting up the ad- and desorption half cycle into a transient, strongly non-isothermal switching phase and a quasi-isothermal phase. In the quasi-isothermal phase, the heat flow rates can be calculated with relationship between temperature effectiveness (ϵ) and number of transfer units (NTU). Effective thermal resistances account for the heat and mass transfer processes. The prediction quality of our simple calculation in terms of heat flow rates is within ±20% compared with experimental data of two different sorption modules. The suggested method and its experimental validation lay the foundation of a basic adsorption heat exchanger theory.


Comparative anti-microbial efficacy of Azadirachta indica irrigant with standard endodontic irrigants: A preliminary study.
The anti-microbial efficacy of 2.5% sodium hypochlorite (SHC) and 0.2% chlorhexidine gluconate were compared with an experimental irrigant formulated from the Neem tree, Azadirachta indica A. Juss. A sample of 36 single rooted anterior teeth with periapical radiolucency and absence of response to vitality tests that required root canal treatment were selected for this study. The test irrigants and their combinations were assigned to five different groups and saline served as the control. Access cavities were prepared using an aseptic technique and samples collected for both anaerobic culture and Gram stained smears, followed by irrigation and sample collection again. The number of organisms were expressed in colony forming units/ml after 72 h of incubation; the smears were analyzed for their microbial loads and tissue clearance and assessed as per defined criteria. Our results found the maximum reduction in microbial loads, when analyzed by culture method, with a combination of SHC and the experimental neem irrigant. Maximum tissue clearance on the Gram Stained smears was also found with the same combination. Neem irrigant has anti-microbial efficacy and can be considered for endodontic use.


Dysmenorrhoea and menstrual abnormalities among postmenarcheal secondary school girls in Maiduguri Nigeria.
Four Hundred and fifteen post menarcheal secondary school girls selected randomly from six secondary schools in Maiduguri metropolis were interviewed with the aid of questionnaires to find out their ages at menarche and associated menstrual problems, regularity of menstrual cycle, menorrhagia and dysmenorrhoea within the first two years following menarche. Their ages ranged between 12 and 21 years. Twenty six students were three months post menarche, 187 were 4-12 months post menarche.'The mean menarcheal age was 13.6 years. Menorrhagia was a very rare form of menstrual abnormality. Early menarche especially between the ages of 12 to 14 years was found to be associated with higher frequency of irregular menstrual cycles while this improves with late onset menarche. About 45% had one form of menstrual abnormality, amenorrhoea (4.6%), oligomenorrhoea (18%) and polymenorrhoea (21%). These were almost uniformly associated with all groups. Over 80% had attained menarche by age of 14 years. Dysmenorrhoea was just slightly commoner by the ages of 15 and 16 years when it occurs more frequently.


Individuals with the dominant hand affected following stroke demonstrate less impairment than those with the nondominant hand affected.
The purpose was to determine if upper extremity impairment and function in individuals with chronic stroke is dependent upon whether the dominant or non-dominant hand is affected. Ninety-three community-dwelling individuals with stroke. The Modified Ashworth Scale (tone), handheld dynamometry (isometric strength), monofilaments (sensation), Brief Pain Inventory (pain), Chedoke Arm and Hand Activity Inventory Motor Activity Log (paretic arm use), and Reintegration to Normal Living Index (participation) were used to form impairment and function models. Multivariate analysis models (Dominance x Severity) were created for impairment and function variables. There was a significant interaction and main effect of Dominance for the impairment model (P = 0.01) but not the function model (P = 0.75). The dependent variables of tone, grip strength, and pain were all significantly affected by Dominance, indicating less impairment if the dominant hand was affected. All dependent variables except pain were affected by Severity. This study looked at the effect of the dominant hand being affected versus the nondominant in individuals with chronic stroke. Individuals with the dominant hand affected demonstrated less impairment than those with the nondominant hand affected. However, there was no effect of dominance on paretic arm use or performance in activities of daily living. Prospective studies to further explore the issue of hand dominance and poststroke function are suggested.


Effects of oral administration of extracts of Hypericum perforatum (St John's wort) on brain serotonin transporter, serotonin uptake and behaviour in mice.
The pharmacological effects of extracts of Hypericum perforatum (St John's wort) were characterized in-vitro and ex-vivo, in relation to its behavioural effects. In in-vitro experiments, St John's wort inhibited brain synaptosomal [(3)H]serotonin uptake in mice with little effect on specific [(3)H]paroxetine binding. For selective serotonin-reuptake inhibitors (SSRIs), the IC50 value for [(3)H]serotonin uptake (molar concentration of unlabelled drug necessary to displace 50% of specific uptake) correlated well with the inhibition constant K(i) value for [(3)H]paroxetine binding in mouse brain. Oral administration of St John's wort (900 mg kg(-1)), paroxetine (1 mg kg(-1)) and sertraline (10 mg kg(-1)) brought about significant increases in the K(m) value for [(3)H]serotonin uptake into brain synaptosomes 4 h later, and only SSRIs suppressed specific [(3)H]paroxetine binding in mouse brain. St John's wort and SSRIs significantly inhibited marble-burying behaviour in mice and the time-course of attenuation of this behaviour by St John's wort was similar to that of [(3)H]serotonin uptake inhibition. In the forced swimming test, St John's wort, but not SSRIs, suppressed the immobility time of mice after oral administration. These results provide the first in-vivo evidence to suggest that the mode of antidepressant action of St John's wort differs from that of SSRIs. Thus, this study may have a significant impact on phytotherapy with St John's wort.


Nasendoscopy for unusual nasal symptoms.
A 20-year-old woman who was fit and well presented with a history of left nasal blockage for 2 years. She was noted on anterior rhinoscopy to have nasal septal deviation towards the left. She was listed for septoplasty with the aim of relieving nasal obstruction. At operation she was found to have a mildly deviated septum to the left. There was also a rhinolith in the left nostril posterior to the deviated septum (figure 1). Following removal of the rhinolith, her nasal airway appeared adequate; hence, septoplasty was not performed. Postoperatively, the patient was pleased with the outcome. When the patient was shown the foreign body she recalled inserting a pen cover into her nose about 10 years previously (figure 2). When she presented to the Accident and Emergency department at that time she was told that there was no foreign body in her nose.


The effects of bilateral movement training on upper limb function in chronic stroke patients.
[Purpose] This study compared the functional and kinematic changes associated with two rehabilitation protocols: bilateral and unilateral movement training. [Subjects and Methods] Twenty-five patients with chronic stroke were randomly assigned to two training protocols for four weeks of training. Each training session consisted of three tasks. The tasks were performed with either the impaired and unimpaired arms moving synchronously (bilateral training) or with the impaired arm alone (unilateral training). To compare the changes associated with each rehabilitation protocol, functional and kinematic assessments were performed before and after the interventions. The functional state of each patient was measured by the Box and Block Test, and the kinematic variables were assessed by three-dimensional motion analysis. The Box and Block Test was used to assess the functional abilities of the affected upper limb. Kinematic measurements of upper limb movement were measured with a 3-dimensional motion analysis system. [Results] Results showed that the bilateral movement group had significantly improved motion of the shoulder compared to the unilateral movement group. [Conclusion] Bilateral movement training should be used to improve upper limb function in patients with chronic stroke.


Alteration of Immunoregulatory Patterns and Survival Advantage of Key Cell Types in Food Allergic Children.
All allergic responses to food indicate the failure of immunological tolerance, but it is unclear why cow's milk and egg (CME) allergies resolve more readily than reactivity to peanuts (PN). We sought to identify differences between PN and CME allergies through constitutive immune status and responses to cognate and non-cognate food antigens. Children with confirmed allergy to CME (n = 6) and PN (n = 18) and non-allergic (NA) (n = 8) controls were studied. Constitutive secretion of cytokines was tested in plasma and unstimulated mononuclear cell (PBMNC) cultures. Blood dendritic cell (DC) subsets were analyzed alongside changes in phenotypes and soluble molecules in allergen-stimulated MNC cultures with or without cytokine neutralization. We observed that in allergic children, constitutively high plasma levels IL-1β, IL-2, IL-4, IL-5 and IL-10 but less IL-12p70 than in non-allergic children was accompanied by the spontaneous secretion of sCD23, IL-1β, IL-2, IL-4, IL-5, IL-10, IL-12p70, IFN-γ and TNF-α in MNC cultures. Furthermore, blood DC subset counts differed in food allergy. Antigen-presenting cell phenotypic abnormalities were accompanied by higher B and T cell percentages with more Bcl-2 within CD69+ subsets. Cells were generally refractory to antigenic stimulation in vitro, but IL-4 neutralization led to CD152 downregulation by CD4+ T cells from PN allergic children responding to PN allergens. Canonical discriminant analyses segregated non-allergic and allergic children by their cytokine secretion patterns, revealing differences and areas of overlap between PN and CME allergies. Despite an absence of recent allergen exposure, indication of in vivo activation, in vitro responses independent of challenging antigen and the presence of unusual costimulatory molecules suggest dysregulated immunity in food allergy. Most importantly, higher Bcl-2 content within key effector cells implies survival advantage with the potential to mount abnormal responses that may give rise to the manifestations of allergy. Here, we put forward the hypothesis that the lack of apoptosis of key immune cell types might be central to the development of food allergic reactions.


Dynamic extension assist splinting of acute central slip lacerations.
The management of zone III extensor tendon injuries remains a challenging clinical problem for both the surgeon and therapist. Current treatment protocols include prolonged immobilization, controlled mobilization, or limited early dynamic splinting, but overall results are often less than optimal. This retrospective study describes an alternative treatment approach for acute central slip lacerations consisting of initial immobilization of the PIP joint in absolute 0 degrees extension for an average of 3.5 weeks followed by active flexion of the PIP joint in a finger-based dynamic extension assist splint. Therapy averaged approximately one visit weekly, and follow-up averaged 3.8 months. Mean PIP joint extension was + 1 degrees, and mean PIP joint flexion was 109 degrees. Mean DIP joint extension was -1 degree and mean DIP flexion was 67 degrees. Mean total active motion measured in composite fashion for the PIP and DIP joints was 175 degrees. This treatment approach produced excellent results, required infrequent therapy, and utilized only finger-based splinting.


Utility of specimens positive for Neisseria gonorrhoeae by the Aptima Combo 2 assay for assessment of strain diversity and antibiotic resistance.
In our jurisdiction, the Aptima Combo 2 assay (Gen-Probe, Inc.) is used to detect Neisseria gonorrhoeae from specimens collected at clinics for sexually transmitted infections (STI) and from select community patients. In addition, swabs are also collected for N. gonorrhoeae culture, susceptibility testing, and sequence typing (ST). Since only a small proportion of samples from provincial cases undergo culture, the available trends in antimicrobial susceptibility and predominant strain types may not be representative of all N. gonorrhoeae infections. Due to the limitations facing the use of N. gonorrhoeae culture to understand these trends in the general community, we performed a molecular analysis for markers of cephalosporin resistance and ST determination by using nucleic acid extracts of specimens sent for Aptima testing. Thirty-four samples submitted for both Aptima testing and N. gonorrhoeae culture from the same anatomic location (within 24 h) were included in the study. Sequence type was determined based on the sequence of the por and tbpB genes, and amino acid changes in the PBP 2 protein, encoded by the penA gene, were considered representative for the assessment of antimicrobial susceptibility. Sequence identity of 100% was observed between the sequences obtained from Aptima-analyzed samples and culture samples. Sequencing results showed an association between decreased susceptibility to extended-spectrum cephalosporins (ESC(ds)), tbp allele 110, ST 1407, and amino acid changes (G545S, I312M, and V316T) in the PBP 2 protein. Our data, generated based on a few representative genes, suggest that gonococcal samples positive by Aptima testing can be used to determine single nucleotide polymorphisms associated with ESC(ds) and the sequence type based on molecular strain typing. Confirmation of these findings may obviate the need for gonorrhea culture in the future.


Hepatitis B: modern concepts in pathogenesis--APOBEC3 cytidine deaminases as effectors in innate immunity against the hepatitis B virus.
APOBEC3 editing enzymes inhibit retroviruses by cytidine deamination in minus-strand cDNA, leading to G to A hypermutated proviruses, and by less well characterized inhibition of retroviral replication independently of catalysis. This review focuses on the effects of APOBEC3 enzymes on the pararetrovirus hepatitis B virus. The cytidine deaminases APOBEC3B, APOBEC3C, APOBEC3F and APOBEC3G deaminate cytidine residues in hepatitis-B-virus minus-strand cDNA, resulting in G to A hypermutated genomes in the serum of hepatitis-B-virus-infected patients. APOBEC3B, APOBEC3F and APOBEC3G directly inhibit hepatitis-B-virus reverse transcription independently of deaminase activity. In human liver, APOBEC3B, APOBEC3F and APOBEC3G are expressed to low levels, but in human primary hepatocytes stimulated with interferon-alpha, APOBEC3G is induced to levels sufficient for hepatitis-B-virus inhibition. APOBEC3B inhibits hepatitis-B-virus gene transcription, and APOBEC3B and APOBEC3G preferentially mutate the hepatitis-B-virus x gene leading to the truncated hepatitis-B-virus x variants in hepatitis-B-virus-associated liver cancer. The interferon-inducible APOBEC3G and the other APOBEC3s restrict hepatitis B virus by cytidine deamination in hepatitis-B-virus minus-strand cDNA and by direct inhibition of hepatitis-B-virus reverse transcriptase. The nuclear localized APOBEC3B is implicated in liver cancer development. To what extent these enzymes contribute to noncytolytic clearance of hepatitis B virus in vivo remains to be defined, yet the APOBEC3 cytidine deaminases are likely to play a role in these processes.


How trust in institutions and organizations builds general consumer confidence in the safety of food: a decomposition of effects.
This paper investigates the relationship between general consumer confidence in the safety of food and consumer trust in institutions and organizations. More specifically, using a decompositional regression analysis approach, the extent to which the strength of the relationship between trust and general confidence is dependent upon a particular food chain actor (for example, food manufacturers) is assessed. In addition, the impact of specific subdimensions of trust, such as openness, on consumer confidence are analyzed, as well as interaction effects of actors and subdimensions of trust. The results confirm previous findings, which indicate that a higher level of trust is associated with a higher level of confidence. However, the results from the current study extend on previous findings by disentangling the effects that determine the strength of this relationship into specific components associated with the different actors, the different trust dimensions, and specific combinations of actors and trust dimensions. The results show that trust in food manufacturers influences general confidence more than trust in other food chain actors, and that care is the most important trust dimension. However, the contribution of a particular trust dimension in enhancing general confidence is actor-specific, suggesting that different actors should focus on different trust dimensions when the purpose is to enhance consumer confidence in food safety. Implications for the development of communication strategies that are designed to regain or maintain consumer confidence in the safety of food are discussed.


Doppler-derived intracoronary physiology indices predict the occurrence of microvascular injury and microvascular perfusion deficits after angiographically successful primary percutaneous coronary intervention.
A total of 40% to 50% of patients with ST-segment-elevation myocardial infarction develop microvascular injury (MVI) despite angiographically successful primary percutaneous coronary intervention (PCI). We investigated whether hyperemic microvascular resistance (HMR) immediately after angiographically successful PCI predicts MVI at cardiovascular magnetic resonance and reduced myocardial blood flow at positron emission tomography (PET). Sixty patients with ST-segment-elevation myocardial infarction were included in this prospective study. Immediately after successful PCI, intracoronary pressure-flow measurements were performed and analyzed off-line to calculate HMR and indices derived from the pressure-velocity loops, including pressure at zero flow. Cardiovascular magnetic resonance and H2 (15)O PET imaging were performed 4 to 6 days after PCI. Using cardiovascular magnetic resonance, MVI was defined as a subendocardial recess of myocardium with low signal intensity within a gadolinium-enhanced area. Myocardial perfusion was quantified using H2 (15)O PET. Reference HMR values were obtained in 16 stable patients undergoing coronary angiography. Complete data sets were available in 48 patients of which 24 developed MVI. Adequate pressure-velocity loops were obtained in 29 patients. HMR in the culprit artery in patients with MVI was significantly higher than in patients without MVI (MVI, 3.33±1.50 mm Hg/cm per second versus no MVI, 2.41±1.26 mm Hg/cm per second; P=0.03). MVI was associated with higher pressure at zero flow (45.68±13.16 versus 32.01±14.98 mm Hg; P=0.015). Multivariable analysis showed HMR to independently predict MVI (P=0.04). The optimal cutoff value for HMR was 2.5 mm Hg/cm per second. High HMR was associated with decreased myocardial blood flow on PET (myocardial perfusion reserve <2.0, 3.18±1.42 mm Hg/cm per second versus myocardial perfusion reserve ≥2.0, 2.24±1.19 mm Hg/cm per second; P=0.04). Doppler-flow-derived physiological indices of coronary resistance (HMR) and extravascular compression (pressure at zero flow) obtained immediately after successful primary PCI predict MVI and decreased PET myocardial blood flow. http://www.trialregister.nl. Unique identifier: NTR3164.


Association Between S100b Levels and COVID-19 Pneumonia: A Case Control Study.
Extracellular S100b effects are mediated by the receptor for advanced glycation end products (RAGE), which is the S100b membrane receptor. RAGE belongs to the immunoglobulin superfamily of cell surface molecules and serves as a multiligand receptor and is expressed in high abundance by alveolar type I (AT-I) cells in adult pulmonary tissue. This study aimed to provide an insight into the association between the severity of COVID-19 disease and serum S100b levels during admission to the emergency department (ED). A total of 64 patients (34 mild cases; 30 severe cases) were diagnosed with COVID-19 pneumonia and 30 healthy volunteers were admitted to study. Serum S100b levels were measured by using enzymle linked immunoassay method from blood serum samples. Serum S100b levels showed a significantly higher mean value in mild and severe disease cohorts than in healthy controls (p=0.036 and p=0.028 respectively). Receiver operating characteristic (ROC) analysis indicated greater area under the curve (AUC) for serum S100b levels of the COVID-19 patients (AUC=0.663, 95% CI=0.541-0.785; p=0.014). In addition, serum S100b concentration was measured as 151.7 ng/ml at 79.3% sensitivity and 51.7% specificity (p=0.014). Serum S100b protein levels can serve as a valuable clinical marker in establishing diagnosis of patients. Though not useful in identifying different stages of COVID-19 infection, serum S100b concentration along with other known markers can be utilized to reliably predict clinical severity along with other clinical parameters.


Opiate receptor function may be modulated through an oxidation-reduction mechanism.
Cupric ion, a thiol oxidant, caused naloxone-reversible analgesia when injected intracerebroventricularly in mice; its potency was close to that of morphine. Dithiothreitol, a thiol reductant, reversed the analgesia induced by cupric ion and antagonized analgesia induced by morphine. Oxidized dithiothreitol had no effect. These findings, together with evidence for redox modification of opiate receptor binding in vitro, suggest that a mechanism of oxidation-reduction of thiols may modulate opiate receptor function.


Cluster of genes controlling proline degradation in Salmonella typhimurium.
A cluster of genes essential for degradation of proline to glutamate (put) is located between the pyrC and pyrD loci at min 22 of the Salmonella chromosome. A series of 25 deletion mutants of this region have been isolated and used to construct a fine-structure map of the put genes. The map includes mutations affecting the proline degradative activities, proline oxidase and pyrroline-5-carboxylic dehydrogenase. Also included are mutations affecting the major proline permease and a regulatory mutation that affects both enzyme and permease production. The two enzymatic activities appear to be encoded by a single gene (putA). The regulatory mutation maps between the putA gene and the proline permease gene (putP).


5-Fluorouracil incorporation into RNA and DNA in relation to thymidylate synthase inhibition of human colorectal cancers.
The mechanism of action of 5-fluorouracil (5-FU) has been associated with inhibition of thymidylate synthase (TS) and incorporation of 5-FU into RNA and DNA, but limited data are available in human tumor tissue for the latter. We therefore measured incorporation in human tumor biopsy specimens after administration of a test dose of 5-FU alone or with leucovorin. Patients received 5-FU (500 mg/m(2)) with or without high-dose leucovorin, low-dose leucovorin or l-leucovorin, and biopsy specimens were taken after approximately 2, 24 or 48 h. Tissues were pulverized and extracted for nucleic acids. 5-FU incorporation was measured using gas chromatography/mass spectrometry after complete degradation to bases of isolated RNA and DNA. Maximal incorporation into RNA (1.0 pmol/micrograms RNA) and DNA (127 fmol/micrograms DNA) of 59 and 46 biopsy specimens, respectively, was found at 24 h after 5-FU administration. Incorporation into RNA but not DNA was significantly correlated with intratumoral 5-FU levels. However, DNA incorporation was significantly correlated with the RNA incorporation. Primary tumor tissue, liver metastasis and normal mucosa did not show significant differences, while leucovorin had no effect. Neither for RNA (30 patients) nor DNA (24 patients) incorporation was a significant correlation with response to 5-FU therapy found. However, in the same group of patients, response was significantly correlated to TS inhibition (mean TS in responding and non-responding groups 45 and 231 pmol/h/mg protein, respectively; P=0.001). 5-FU is incorporated at detectable levels into RNA and DNA of human tumor tissue, but no relation between the efficacy of 5-FU treatment and incorporation was found, in contrast to TS.


Effects of vitamin B6 on averaged evoked potentials in infantile autism.
In autistic children, averaged evoked potentials have been reported to have lower amplitudes and shorter latencies than those of normal children. Also, moderate clinical improvement has been observed in some autistic children after treatment with vitamin B6 and magnesium. We have studied biochemical and electrophysiological effects of vitamin B6 and magnesium in 12 autistic children and in 11 normal children. During treatment of the autistic children with B6, an increase of amplitude of middle-latency evoked potentials and a decrease of urinary homovanillic acid were found. The reverse was noted in the normal subjects.


Apoplastic polyamine oxidation plays different roles in local responses of tobacco to infection by the necrotrophic fungus Sclerotinia sclerotiorum and the biotrophic bacterium Pseudomonas viridiflava.
The role of polyamine (PA) metabolism in tobacco (Nicotiana tabacum) defense against pathogens with contrasting pathogenic strategies was evaluated. Infection by the necrotrophic fungus Sclerotinia sclerotiorum resulted in increased arginine decarboxylase expression and activity in host tissues, as well as putrescine and spermine accumulation in leaf apoplast. Enhancement of leaf PA levels, either by using transgenic plants or infiltration with exogenous PAs, led to increased necrosis due to infection by S. sclerotiorum. Specific inhibition of diamine and PA oxidases attenuated the PA-induced enhancement of leaf necrosis during fungal infection. When tobacco responses to infection by the biotrophic bacterium Pseudomonas viridiflava were investigated, an increase of apoplastic spermine levels was detected. Enhancement of host PA levels by the above-described experimental approaches strongly decreased in planta bacterial growth, an effect that was blocked by a PA oxidase inhibitor. It can be concluded that accumulation and further oxidation of free PAs in the leaf apoplast of tobacco plants occurs in a similar, although not identical way during tobacco defense against infection by microorganisms with contrasting pathogenesis strategies. This response affects the pathogen's ability to colonize host tissues and results are detrimental for plant defense against necrotrophic pathogens that feed on necrotic tissue; on the contrary, this response plays a beneficial role in defense against biotrophic pathogens that depend on living tissue for successful host colonization. Thus, apoplastic PAs play important roles in plant-pathogen interactions, and modulation of host PA levels, particularly in the leaf apoplast, may lead to significant changes in host susceptibility to different kinds of pathogens.


Co-Delivery of Bioengineered Exosomes and Oxygen for Treating Critical Limb Ischemia in Diabetic Mice.
Diabetic patients with critical limb ischemia face a high rate of limb amputation. Regeneration of the vasculature and skeletal muscles can salvage diseased limbs. Therapy using stem cell-derived exosomes that contain multiple proangiogenic and promyogenic factors represents a promising strategy. Yet the therapeutic efficacy is not optimal because exosomes alone cannot efficiently rescue and recruit endothelial and skeletal muscle cells and restore their functions under hyperglycemic and ischemic conditions. To address these limitations, we fabricated ischemic-limb-targeting stem cell-derived exosomes and oxygen-releasing nanoparticles and codelivered them in order to recruit endothelial and skeletal muscle cells, improve cell survival under ischemia before vasculature is established, and restore cell morphogenic function under high glucose and ischemic conditions. The exosomes and oxygen-releasing nanoparticles, delivered by intravenous injection, specifically accumulated in the ischemic limbs. Following 4 weeks of delivery, the exosomes and released oxygen synergistically stimulated angiogenesis and muscle regeneration without inducing substantial inflammation and reactive oxygen species overproduction. Our work demonstrates that codelivery of exosomes and oxygen is a promising treatment solution for saving diabetic ischemic limbs.


Huge Rashba-type spin-orbit coupling in binary hexagonal PX nanosheets (X = As, Sb, and Bi).
We theoretically propose that Rashba-type band splitting can be achieved in binary alloyed hexagonal PX nanosheets (X = As, Sb, and Bi). The lack of inversion symmetry results in an effective electric field perpendicular to the basal plane of PX, hence, leading to Rashba-type spin-orbit coupling (SOC) in the two dimensional PX nanosheets. Since the SOC strength roughly scales quadratically with atomic number, the largest band splitting is found in PBi with a Rashba coefficient of ∼1.56 eV Å, which is a huge value among two-dimensional materials. Furthermore, tensile biaxial strain can be employed to significantly enhance the strength of SOC, for instance, a Rashba coefficient of 4.41 eV Å can be realized at a strain of 10%. The huge and strain-tunable Rashba-type SOC of PBi suggests that it holds great promise for spintronic applications.


Halothane changes the relationships between lung resistances and lung volume.
The authors hypothesized that relaxation of airway smooth muscle by halothane lessens the dependence of airway resistance on lung volume, and that halothane alters the relationship between pulmonary resistance and lung volume by changing both the airway and tissue components of pulmonary resistance. The relationship among airway resistance, tissue resistance, and lung volume was examined in mongrel dogs before and during the administration of halothane, both in airways with reduced smooth muscle tone (after vagotomy) and during moderate increases in smooth muscle tone caused by vagus nerve stimulation (VNS). Resistance were measured at several levels of positive end-expiratory pressure (PEEP, 4-15 cmH2O) using an alveolar capsule technique. Before halothane administration, airway resistance increased at low PEEP; VNS accentuated this increase. Tissue resistance increased at low PEEP only during VNS. Halothane had no significant effect on any resistance before VNS. During VNS, halothane markedly blunted increases in airway resistance and tissue resistance as PEEP decreased. The authors conclude that during stimulation of airway smooth muscle in dogs, halothane attenuates increases in airway resistance and tissue resistance with reductions in lung volume in dogs. Thus, moderate changes in lung volume have little effect on these resistances during halothane anesthesia under these conditions.


Communication Tools for End-of-Life Decision-Making in Ambulatory Care Settings: A Systematic Review and Meta-Analysis.
Patients with serious illness, and their families, state that better communication and decision-making with healthcare providers is a high priority to improve the quality of end-of-life care. Numerous communication tools to assist patients, family members, and clinicians in end-of-life decision-making have been published, but their effectiveness remains unclear. To determine, amongst adults in ambulatory care settings, the effect of structured communication tools for end-of-life decision-making on completion of advance care planning. We searched for relevant randomized controlled trials (RCTs) or non-randomized intervention studies in MEDLINE, EMBASE, CINAHL, ERIC, and the Cochrane Database of Randomized Controlled Trials from database inception until July 2014. Two reviewers independently screened articles for eligibility, extracted data, and assessed risk of bias. Grading of Recommendations Assessment, Development, and Evaluation (GRADE) was used to evaluate the quality of evidence for each of the primary and secondary outcomes. Sixty-seven studies, including 46 RCTs, were found. The majority evaluated communication tools in older patients (age >50) with no specific medical condition, but many specifically evaluated populations with cancer, lung, heart, neurologic, or renal disease. Most studies compared the use of communication tools against usual care, but several compared the tools to less-intensive advance care planning tools. The use of structured communication tools increased: the frequency of advance care planning discussions/discussions about advance directives (RR 2.31, 95% CI 1.25-4.26, p = 0.007, low quality evidence) and the completion of advance directives (ADs) (RR 1.92, 95% CI 1.43-2.59, p<0.001, low quality evidence); concordance between AD preferences and subsequent medical orders for use or non-use of life supporting treatment (RR 1.19, 95% CI 1.01-1.39, p = 0.028, very low quality evidence, 1 observational study); and concordance between the care desired and care received by patients (RR 1.17, 95% CI 1.05-1.30, p = 0.004, low quality evidence, 2 RCTs). The use of structured communication tools may increase the frequency of discussions about and completion of advance directives, and concordance between the care desired and the care received by patients. The use of structured communication tools rather than an ad-hoc approach to end-of-life decision-making should be considered, and the selection and implementation of such tools should be tailored to address local needs and context. PROSPERO CRD42014012913.


Adaptation to display rotation and display gain distortions during drawing.
The relationship between movement extent and movement direction coding mechanisms was investigated using a visuomotor adaptation paradigm. To determine if these mechanisms are either modular or interdependent, young healthy college students were tested while they performed a visually guided drawing task that incorporated varying combinations of movement distance and direction distortions. Analysis of participants' standardized movement duration, initial directional error, and movement length over the course of the adaptation process revealed a certain degree of interdependence between direction and extent coding mechanisms. Specifically, changes in final adaptation levels and after-effects depended on the order of introduction of the visual distortions. This interaction can be characterized as unidirectional, where alterations in rotational feedback interfere with subsequent adaptation to gain changes, whereas alterations in "display gain" do not significantly impede the adaptation to "display rotation". Moreover, simultaneous exposure to gain and rotational distortions resulted in better learning. The results argue against an independent coding of movement direction and extent during adaptation by the central nervous system.


Ileal "J" pouch-anal anastomosis. Clinical outcome.
One hundred eighty-eight patients undergoing abdominal colectomy with distal mucosal proctectomy and endorectal ileal pouch-anal anastomosis were reviewed to assess long-term functional results and to identify factors that might influence them. There was no postoperative mortality, but 10 patients (5.3%) required permanent ileostomy because of postoperative complications or the development of unsuspected Crohn's disease. Immediate postoperative complications, including pelvic sepsis, small bowel obstruction requiring surgery, anastomotic stricture, and ileostomy dysfunction, were observed in 11%, 9%, 14% and 9% of patients, respectively. No males were impotent but nine (9%) developed retrograde ejaculation. Pouchitis occurred in 8% of patients. Among 157 patients assessed at least 60 days after ileostomy closure (mean +/- SD, 375 +/- 216 days), all evacuated their neorectum spontaneously, and stool frequency was 6.0 +/- 2.6 daily and 1.2 +/- 1.3 nightly. While continence was generally good, 2.5% of patients during waking hours and 4.5% during sleep had occasional frank soilage. Moreover, seepage was noted in 25 and 47% of patients during daytime and nighttime, respectively. Both stool frequency and degree of continence improved with time. Patients less than 50 years of age and those with polyposis coli had fewer stools and better continence than those older than 50 or those with ulcerative colitis. It is concluded that ileal "J" pouch-anal anastomosis can be performed safely and will provide acceptable anorectal function without late deterioration.


Discovery of highly specific protein markers for the identification of biological stains.
DNA profiling has transformed the field of forensic biology by making it possible to individualize biological stains. The identification of the stain itself, however, continues to present forensic serologists with significant challenges. Current antibody- and enzyme activity-based assays yield only presumptive results as detection in nontarget body fluids or cross-reactivity with nonhuman sources have both been well documented. For other critical body fluids such as vaginal and menstrual fluids, there are no commercial tests at all. Using a three-pronged, comparative proteomic strategy based on proteome fractionation by HPLC followed by MS, a panel of 29 candidate protein biomarkers have been proposed as highly specific indicators of human saliva, urine, seminal fluid, vaginal fluid, peripheral blood, and menstrual fluid. The combination of consistent identification by multiple strategies in the current study; confirmation in independently compiled proteomic databases; and information on tissue expression and/or functionality from the proteomic literature all support the proposition that these proteins will have utility as reliable biomarkers of their target body fluids. The identification of candidate high-specificity protein biomarkers for human body fluids encountered in forensic investigations lays the foundation for the development of faster and more reliable approaches to the serological analysis of evidentiary stains.


An experiment-based model for the petrogenesis of high-alumina basalts.
To understand magmatism at convergent margins, one must know the origin of their characteristic, plagioclase-rich, high-alumina basalts (HABs). Wet melting experiments on basalts at 3 kilobars yielded high-alumina liquids and a coexisting mineral assemblage with little or no plagioclase. An isothermal pressure drop to 1 kilobar caused 20 to 30 percent plagioclase crystallization in these melts, while mafic minerals underwent limited crystallization or even resorption. These results suggest that hydrous (>/=4 percent H(2)O) HAB liquids, presumably formed by fractionation of a hydrous basalt at depth, will precipitate voluminous plagioclase as pressure drops during ascent and eruption. Plagioclase accumulation is not necessarily required to explain the petrogenesis of plagioclase-rich HAB.


Limelight on alpha-synuclein: pathological and mechanistic implications in neurodegeneration.
The pathogenesis of many neurodegenerative disorders arises in association with the misfolding and accumulation of a wide variety of proteins. Much emphasis has been placed on understanding the nature of these protein accumulations, including their composition, the process by which they are formed and the physiological impact they impose at cellular and, ultimately, organismal levels. Alpha-synuclein (ASYN) is the major component of protein inclusions known as Lewy bodies and Lewy neurites, which are the typical pathological hallmarks in disorders referred to as synucleinopathies. In addition, mutations or multiplications in the gene encoding for ASYN have also been shown to cause familial cases of PD, the most common synucleinopathy. Although the precise function of ASYN remains unclear, it appears to be involved in a vast array of cellular processes. Here, we review, in depth, a spectrum of cellular and molecular mechanisms that have been implicated in synucleinopathies. Importantly, detailed understanding of the biology/pathobiology of ASYN may enable the development of novel avenues for diagnosis and/or therapeutic intervention in synucleinopathies.


DOSE: an R/Bioconductor package for disease ontology semantic and enrichment analysis.
Disease ontology (DO) annotates human genes in the context of disease. DO is important annotation in translating molecular findings from high-throughput data to clinical relevance. DOSE is an R package providing semantic similarity computations among DO terms and genes which allows biologists to explore the similarities of diseases and of gene functions in disease perspective. Enrichment analyses including hypergeometric model and gene set enrichment analysis are also implemented to support discovering disease associations of high-throughput biological data. This allows biologists to verify disease relevance in a biological experiment and identify unexpected disease associations. Comparison among gene clusters is also supported. DOSE is released under Artistic-2.0 License. The source code and documents are freely available through Bioconductor (http://www.bioconductor.org/packages/release/bioc/html/DOSE.html). Supplementary data are available at Bioinformatics online. email@example.com or email@example.com.


[Anesthetic management in mastocytosis].
Mastocytosis is a heterogeneous grouping of entities characterized by proliferation of mast cells in one or more organs or tissues. The skin is the most frequently affected organ, followed by bone marrow. The relevance of this disease to anesthesia lies in the fact that many drugs used can trigger massive release of chemical mediators of mast cells. We report the case of a patient diagnosed with cutaneous mastocytosis who underwent testicular biopsy with intradural anesthesia. We review the fundamental aspects of the disease and the principles of anesthetic management.


Prevalence of and Risk Factors for Drug-Related Readmissions in Older Adults: A Systematic Review and Meta-Analysis.
Older adults are at an increased risk of drug-related problems, especially following discharge from hospital. Drug-related readmissions place a large burden on the patient and the healthcare system. However, previous studies report inconsistent results on the prevalence and associated risk factors for drug-related hospital readmissions in older adults. We aimed to assess the prevalence of drug-related readmissions in older adults aged 65 years and older and investigate the drug classes, preventability and risk factors most associated with these readmissions. A systematic review and meta-analysis were undertaken to answer our objectives. A search of four databases (MEDLINE, Embase, CINAHL and Scopus) was conducted. Three authors independently performed title and abstract screening, full-text screening and data extraction of all included studies. A meta-analysis was conducted to calculate the pooled prevalence of drug-related readmissions across all studies, and a subgroup analysis was performed to explore heterogeneity among studies reporting on adverse drug reaction-related readmissions. A total of 1978 studies were identified in the initial search, of which four studies were included in the final synthesis. Three studies focused on readmissions due to adverse drug reactions and one study focused on readmissions due to drug-related problems. A pooled prevalence of 9% (95% confidence interval 2-18) was found for drug-related readmissions across all studies, and a pooled prevalence of 6% (95% confidence interval 4-10) was found for adverse drug reaction-related readmissions. Three studies explored the preventability of readmissions and 15.4-22.2% of cases were deemed preventable. The drug classes most associated with adverse drug reaction readmissions included anticoagulants, antibiotics, psychotropics and chemotherapy agents. Polypharmacy (the use of five or more medications) and several comorbidities such as cancer, liver disease, ischaemic heart disease and peptic ulcer disease were identified as risk factors for drug-related readmissions. Almost one in ten older adults discharged from hospital experienced a drug-related hospital readmission, with one fifth of these deemed preventable. Several comorbidities and the use of polypharmacy and high-risk drugs were identified as prominent risk factors for readmission. Further research is needed to explore possible causes of drug-related readmissions in older adults for a more guided approach to the development of effective medication management interventions.


Evaluation of E-cigarette liquid vapor and mainstream cigarette smoke after direct exposure of primary human bronchial epithelial cells.
E-cigarettes are emerging products, often described as "reduced-risk" nicotine products or alternatives to combustible cigarettes. Many smokers switch to e-cigarettes to quit or significantly reduce smoking. However, no regulations for e-cigarettes are currently into force, so that the quality and safety of e-liquids is not necessarily guaranteed. We exposed primary human bronchial epithelial cells of two different donors to vapor of e-cigarette liquid with or without nicotine, vapor of the carrier substances propylene glycol and glycerol as well as to mainstream smoke of K3R4F research cigarettes. The exposure was done in a CULTEX® RFS compact module, allowing the exposure of the cells at the air-liquid interface. 24 h post-exposure, cell viability and oxidative stress levels in the cells were analyzed. We found toxicological effects of e-cigarette vapor and the pure carrier substances, whereas the nicotine concentration did not have an effect on the cell viability. The viability of mainstream smoke cigarette exposed cells was 4.5-8 times lower and the oxidative stress levels 4.5-5 times higher than those of e-cigarette vapor exposed cells, depending on the donor. Our experimental setup delivered reproducible data and thus provides the opportunity for routine testing of e-cigarette liquids to ensure safety and quality for the user.


Neural crest determination by co-activation of Pax3 and Zic1 genes in Xenopus ectoderm.
A number of regulatory genes have been implicated in neural crest development. However, the molecular mechanism of how neural crest determination is initiated in the exact ectodermal location still remains elusive. Here, we show that the cooperative function of Pax3 and Zic1 determines the neural crest fate in the amphibian ectoderm. Pax3 and Zic1 are expressed in an overlapping manner in the presumptive neural crest area of the Xenopus gastrula, even prior to the onset of the expression of the early bona fide neural crest marker genes Foxd3 and Slug. Misexpression of both Pax3 and Zic1 together efficiently induces ectopic neural crest differentiation in the ventral ectoderm, whereas overexpression of either one of them only expands the expression of neural crest markers within the dorsolateral ectoderm. The induction of neural crest differentiation by Pax3 and Zic1 requires Wnt signaling. Loss-of-function studies in vivo and in the animal cap show that co-presence of Pax3 and Zic1 is essential for the initiation of neural crest differentiation. Thus, co-activation of Pax3 and Zic1, in concert with Wnt, plays a decisive role for early neural crest determination in the correct place of the Xenopus ectoderm.


[Evaluation of the Mannheim Peritonitis Index].
A retrospective analysis of 100 case histories of patients with diffuse peritonitis was made in order to evaluate the prognostic significance of the Mannheim Peritonitis Index (MPI). The patients were divided into 3 groups according to the amount of scores: in the first group (12-20 scores) there were no lethal issues, in the second group (21-29 scores) 42% of the patients died, 100% lethality was noted in the third group when MPI was 30 scores or more. MPI allows not only to make a retrospective evaluation of results of treatment of peritonitis and compare outcomes of different surgical accesses, it may be one of criteria for programmed relaparotomy or for using laparostomy in patients of the 2nd and 3rd groups.


Crystal structure of the human lamin A coil 2B dimer: implications for the head-to-tail association of nuclear lamins.
Nuclear intermediate filaments (IFs) are made from fibrous proteins termed lamins that assemble, in association with several transmembrane proteins of the inner nuclear membrane and an unknown number of chromatin proteins, into a filamentous scaffold called the nuclear lamina. In man, three types of lamins with significant sequence identity, i.e. lamin A/C, lamin B1 and B2, are expressed. The molecular characteristics of the filaments they form and the details of the assembly mechanism are still largely unknown. Here we report the crystal structure of the coiled-coil dimer from the second half of coil 2 from human lamin A at 2.2A resolution. Comparison to the recently solved structure of the homologous segment of human vimentin reveals a similar overall structure but a different distribution of charged residues and a different pattern of intra- and interhelical salt bridges. These features may explain, at least in part, the differences observed between the lamin and vimentin assembly pathways. Employing a modeled lamin A coil 1A dimer, we propose that the head-to-tail association of two lamin dimers involves strong electrostatic attractions of distinct clusters of negative charge located on the opposite ends of the rod domain with arginine clusters in the head domain and the first segment of the tail domain. Moreover, lamin A mutations, including several in coil 2B, have been associated with human laminopathies. Based on our data most of these mutations are unlikely to alter the structure of the dimer but may affect essential molecular interactions occurring in later stages of filament assembly and lamina formation.


Measuring cohabitation and family structure in the United States: assessing the impact of new data from the Current Population Survey.
In 2007, the Current Population Survey (CPS) introduced a measure that identifies all cohabiting partners in a household, regardless of whether they describe themselves as "unmarried partners" in the relationship to householder question. The CPS now also links children to their biological, step-, and adoptive parents. Using these new variables, we analyze the prevalence of cohabitation as well as the demographic and socioeconomic characteristics of different-sex cohabiting couples during the years 2007-2009. Estimates of cohabitation produced using only unmarried partnerships miss 18 % of all cohabiting unions and 12 % of children residing with cohabiting parents. Although differences between unmarried partners and most newly identified cohabitors are small, newly identified cohabitors are older, on average, and are less likely to be raising shared biological or adopted children. These new measures also allow us to identify a small number of young, disadvantaged couples who primarily reside in households of other family members, most commonly with parents. We conclude with an examination of the complex living arrangements and poverty status of American children, demonstrating the broader value of these new measures for research on American family and household structure.


A RCA-based assay for analyzing individual strand break in DNA heteroduplex cleavage by restriction endonucleases.
We have developed a rapid and high-throughput assay based on rolling circle amplification, to distinguish individual strand cleavage of DNA duplexes by restriction endonucleases. As an illustration, we analyzed nicking activity of Nb.BbvCI and uneven cleavage of LNA modified DNA by EcoRI. This assay has potential for analyzing protein-DNA interactions.


Simultaneous ex vivo expansion of cytomegalovirus and Epstein-Barr virus-specific cytotoxic T lymphocytes using B-lymphoblastoid cell lines expressing cytomegalovirus pp65.
Cytomegalovirus (CMV) infection and Epstein-Barr virus (EBV)-induced lymphoproliferative disease are serious complications associated with allogeneic stem cell transplantation. Immunotherapy using ex vivo expanded, virus-specific cytotoxic T lymphocytes (CTL) has been explored and proven to be effective in therapeutic or prophylactic regimens for CMV and EBV infections. To generate CTL specific for both CMV and EBV, we engineered EBV-transformed B-lymphoblastoid cell lines (BLCL) to express CMV pp65 for use as antigen-presenting cells (APC). BLCL were transduced with a recombinant retrovirus encoding pp65, the immunodominant CMV polypeptide. Western blot analysis and immunocytochemistry confirmed the expression of pp65 in the transduced cells. Peripheral blood mononuclear cells (PBMC) from healthy CMV seropositive donors were stimulated with autologous pp65-expressing BLCL weekly for 3 weeks. Chromium release assays showed that the resulting CTL cultures possessed specific cytotoxicity against EBV and CMV. Recombinant vaccinia viruses encoding individual CMV peptides were used to demonstrate that this CMV-specific cytotoxicity was specific for pp65. Assays on CD4- and CD8-depleted CTL fractions indicated that CD8(+) CTL mediated the pp65-specific cytotoxicity. These CMV/EBV-specific CTL recognized CMV- and EBV-infected targets sharing HLA class I antigens, but not HLA mismatched targets. Our results demonstrate that BLCL can be used as APC to stimulate expansion of EBV- and CMV-specific CTL simultaneously. These findings have potential implications for posttransplant CMV and EBV immunotherapy in recipients of allogeneic stem cell transplants.


DNA polymorphisms of the lipoprotein lipase gene and their association with coronary artery disease in the Saudi population.
Coronary heart disease (CHD) is a major health problem and a major cause of death in most countries. Evidence has been presented that gene polymorphisms (HindIII, PvuII and Ser447Ter) of lipoprotein lipase (LPL) are risk factors of coronary artery disease (CAD). Our objective of the present investigation was to determine whether 3 LPL polymorphisms (LPL-HindIII, LPL-PvuII and LPL-Ser447Ter) can be considered as independent risk factors for CAD in the Saudi population. We recruited 120 CAD subjects, confirmed angiographically with identical ethnic backgrounds and 65 control subjects. Polymerase chain reaction-restriction fragment length polymorphisms (RFLP) technique was used to detect the polymorphisms of the LPL gene. For the HindIII genotype, within the CAD group, the frequencies of the H(+)H(+) were found in 50.8%, whereas 44.2% carried the H(-)H(+) genotype, and 5% carried the H(-)H(-) genotype. Within the control group, the H(+)H(+) genotype was found in 44.6%, whereas 35.4% carried the H(-)H(+) genotype, 20% carried the H(-)H(-) genotype. The odds ratio (OR) of HindIII genotype H(+)H(+)vs. H(-)H(-) genotype at 95% Confidence Interval (CI) were 4.6 (1.57-13.2) and p < 0.005, hence showing no significant association with CAD. For the PvuII genotype, within the CAD group the frequencies of the P(+)P(+) found in 41.7% whereas 43.3.2% carried the P(-)P(+) genotype, and 15% carried the P(-)P(-) genotype. Within the control group the P(+)P(+) was found in 38.5%, 43.0% carried the P(-)P(+) genotype, and 18.5% carried the P(-)P(-) genotype. The OR of PvuII genotype P(+)P(+)vs. P(-)P(-) genotypes (95% CI) is 1.33 and p = 0.52; hence, it was also insignificant to show association with the disease. For the Ser447Ter genotype, within the CAD group, the frequencies of the C/C found in 83.3%, whereas 16.7% carried the C/G genotype. Within the control group, the C/C was found in 87.7% and 12.3% carried the C/G genotype. We did not get any GG genotypes in control as well as patients for this gene. It can be concluded that C allele of gene masks the presence of G allele in the Saudi population. The OR of CG + GG vs. CC (95% CI) is 1.43 from 0.59 to 3.44 which is insignificant. Hence this gene also has no significant association with CAD in the Saudi population.


Antisense transcription: a critical look in both directions.
The mammalian genome contains a large layer of hidden biological information. High-throughput methods have provided new insights into the regulatory networks that orchestrate the "when, where and how" of gene expression, revealing a complex interplay between proteins, regulatory RNAs, and chemical and structural alterations of the genome itself. Naturally occurring antisense transcription has been considered as an important feature in creating transcriptional and hence cellular and organismal complexity. Here, we review the current understanding of the extent, functions and significance of antisense transcription. We critically discuss results from genome-wide studies and documented examples of individual antisense transcripts. So far, the regulatory potential of gene overlaps has been demonstrated only in a few selected cases of experimentally characterized antisense transcripts. Facing the large-scale antisense transcription observed in eukaryotic genomes, it still remains an open challenge to distinguish transcriptional noise from biological function of gene overlapping patterns.


[Effect of surface-active substances on hydroxylase activity of Tieghemella orchidis in a free and immobilized state].
The effect of nonionogenic surface-active substances (SAS) on hydroxylase activity of Tieghemella orchidis in free and polyacrylamide gel (PAAG) immobilized states was studied. It was shown that addition of SAS (1%) to the cultivation medium simultaneously with the inoculation stimulated the filamentous growth of the mycelium. It was also demonstrated that certain SAS (span-80/sorbitanemonooleate/and polyethylene glycol-600-monolaurate) affected beneficially the culture transformation activity. The hydroxylase activity also increased after mycelium preincubation with tweens (1%) and transformation in the presence of span-20 (sorbitane monolaurate) and span-80 (0.3%). The use of the latter substances during gel preincubation with immobilized cells led to the stabilization of transformation activity, increase in the hydroxylation rate, and a change in the yield of hydroxylated products to produce hydrocortisone.


Benzopyrene promotes lung cancer A549 cell migration and invasion through up-regulating cytokine IL8 and chemokines CCL2 and CCL3 expression.
Tobacco-sourced carcinogen including benzopyrene (B[a]P) in lung cancer metastasis has not been fully reported. In this study, lung carcinoma A549 cell line was used to investigate the potential roles of tobacco-sourced B[a]P on cell metastasis and invasion and to assess its underlying mechanism. Effects of tobacco-sourced carcinogen on A549 cell proliferation, metastasis, and invasion were analyzed using MTT assay, Transwell assay, and scratch method, respectively. The effects of tobacco-sourced carcinogen on cytokines and chemokines secretion were detected using enzyme-linked immunosorbent assay. Moreover, correlation between inflammatory factor expression and cancer cell migration and invasion was assessed using siRNA-mediated gene silencing. Data showed that both B[a]P and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone either at high or low dose performed no significant difference on A549 cell proliferation with time increasing. 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone performed no significant difference on A549 cell migration and invasion while B[a]P significantly increased A549 cell migration and invasion compared to the control group (P < 0.05). Consequently, except for IL-6, IL-8, CCL-2, and CCL-3, secretions were significantly increased by B[a]P treatment compared to the control (P < 0.05). Furthermore, when CCL-2 and CCL-3 were silenced, the migrated and invasive A549 cells were significantly decreased compared to the control, respectively (P < 0.05), while silenced IL-8 drastically decreased the migrated and invasive cells compared to the control (P < 0.01). Taken together, this study illustrated that there may be significant correlation between smoking and lung cancer metastasis. B[a]P maybe an excellent contributor for lung cancer metastasis through up-regulating IL-8, CCL-2, and CCL-3 expression.