Comparative genomic analysis of the odorant-binding protein family in 12 Drosophila genomes: purifying selection and birth-and-death evolution.
Chemoreception is a widespread mechanism that is involved in critical biologic processes, including individual and social behavior. The insect peripheral olfactory system comprises three major multigene families: the olfactory receptor (Or), the gustatory receptor (Gr), and the odorant-binding protein (OBP) families. Members of the latter family establish the first contact with the odorants, and thus constitute the first step in the chemosensory transduction pathway. Comparative analysis of the OBP family in 12 Drosophila genomes allowed the identification of 595 genes that encode putative functional and nonfunctional members in extant species, with 43 gene gains and 28 gene losses (15 deletions and 13 pseudogenization events). The evolution of this family shows tandem gene duplication events, progressive divergence in DNA and amino acid sequence, and prevalence of pseudogenization events in external branches of the phylogenetic tree. We observed that the OBP arrangement in clusters is maintained across the Drosophila species and that purifying selection governs the evolution of the family; nevertheless, OBP genes differ in their functional constraints levels. Finally, we detect that the OBP repertoire evolves more rapidly in the specialist lineages of the Drosophila melanogaster group (D. sechellia and D. erecta) than in their closest generalists. Overall, the evolution of the OBP multigene family is consistent with the birth-and-death model. We also found that members of this family exhibit different functional constraints, which is indicative of some functional divergence, and that they might be involved in some of the specialization processes that occurred through the diversification of the Drosophila genus.


The Feasibility and Effectiveness of a New Practical Multidisciplinary Treatment for Low-Back Pain: A Randomized Controlled Trial.
Low-back pain is a major health problem exacerbated by the fact that most treatments are not suitable for self-management in everyday life. Particularly, interdisciplinary programs consist of intensive therapy lasting several weeks. Additionally, therapy components are rarely coordinated regarding reinforcing effects, which would improve complaints in persons with higher pain. This study assesses the effectiveness of a self-management program, firstly for persons suffering from higher pain and secondly compared to regular routines. Study objectives were treated in a single-blind multicenter controlled trial. A total of n = 439 volunteers (age 18-65 years) were randomly assigned to a twelve-week multidisciplinary sensorimotor training (3-weeks-center- and 9-weeks-homebased) or control group. The primary outcome pain (Chronic-Pain-Grade) as well as mental health were assessed by questionnaires at baseline and follow-up (3/6/12/24 weeks, M2-M5). For statistical analysis, multiple linear regression models were used. N = 291 (age 39.7 ± 12.7 years, female = 61.1%, 77% CPG = 1) completed training (M1/M4/M5), showing a significantly stronger reduction of mental health complaints (anxiety, vital exhaustion) in people with higher than those with lower pain in multidisciplinary treatment. Compared to regular routines, the self-management-multidisciplinary treatment led to a clinically relevant reduction of pain-disability and significant mental health improvements. Low-cost exercise programs may provide enormous relief for therapeutic processes, rehabilitation aftercare, and thus, cost savings for the health system.


Solution conformation of the (-)-trans-anti-5-methylchrysene-dG adduct opposite dC in a DNA duplex: DNA bending associated with wedging of the methyl group of 5-methylchrysene to the 3'-side of the modification site.
This paper reports on NMR-molecular mechanics structural studies of the (-)-trans-anti-[MC]dG adduct positioned opposite dC in the sequence context of the d(C1-C2-A3-T4-C5-[MC]G6-C7-T8-A9-C10-C11).d(G12-G13-T14++ +-A15-G16-C17-G18- A19-T20-G21-G22) duplex [designated (-)-trans-anti-[MC]dG.dC 11-mer duplex]. This adduct is derived from the trans addition at C4 of (-)-anti-1(S),2(R)-dihydroxy-3(R),4(S)-epoxy-1,2,3,4-tetrahydro-5-met hylchrysen e [(-)-anti-5-MeCDE] to the N2 position of dG6 in this duplex sequence. The 5-methyl group is located adjacent to the MC(C4) binding site, with these groups juxtaposed in a sterically crowded bay region in the adduct duplex. The 5-methylchrysenyl and the nucleic acid exchangeable and nonexchangeable protons were assigned following analysis of two-dimensional NMR data sets in H2O and D2O buffer solution. The solution structure of the (-)-trans-anti-[MC]dG.dC 11-mer duplex has been determined by incorporating DNA-DNA and carcinogen-DNA proton-proton distances defined by lower and upper bounds deduced from NOESY data sets as restraints in molecular mechanics computations in torsion angle space. The results establish that the [MC]dG6.dC17 base pair and flanking dC5.dG18 and dC7.dG16 base pairs retain Watson-Crick alignments upon adduct formation. The aromatic chrysenyl ring is positioned in the minor groove of a right-handed B-DNA helix and stacks predominantly over the sugar of the dC17 residue across from it on the unmodified complementary strand. The chrysenyl ring points toward the 3'-end of the modified strand with its 5-methyl group inserting between the modified [MC]dG6.dC17 and dC7.dG16 base pairs. The adduct duplex bends by approximately 47 degrees as a result of the wedged insertion of the 5-methyl group from the minor groove face of the duplex. The solution structure of the (-)-trans-anti-[MC] dG.dC 11-mer duplex is compared with that of the corresponding (-)-trans-anti-[BP]dG.dC 11-mer [De los Santos et al. (1992) Biochemistry 31, 5245-5252] in which the [BP]dG adduct is derived from the binding of (-)-anti-BPDE [7(S),8(R)-dihydroxy-9(R),10(S)-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene] to the N2 position in the same DNA sequence context. Although the solution structures of the (-)-trans-anti-stereoisomers of 5-methylchrysenyl-dG and benzo[a]pyrenyl-dG adducts opposite dC exhibit many features in common with each other, the [MC]dG adduct which contains a bay region methyl group bends the DNA helix to a greater extent than in the corresponding [BP]dG adduct, which lacks a bay region methyl group.(ABSTRACT TRUNCATED AT 400 WORDS)


Solubilization and assay of a colony-stimulating factor receptor from murine macrophages.
The colony-stimulating factor, CSF-1, selectively stimulates the survival, proliferation, and differentiation of mononuclear phagocytes. The solubilization, assay, and characteristics of the CSF-1 receptor from the J774.2 murine macrophage cell line are described. The recovery of cell-surface receptor in the postnuclear supernatant membrane fraction of hypotonically disrupted cells was 76%. Recovery of the ligand binding activity of the receptor after solubilization of this fraction with 1% Triton X-100 was approximately 150%. The binding of 125I-CSF-1 to intact cells and membrane preparations was consistent with the existence of a single class of high-affinity receptor sites. In contrast, the equilibrium binding of 125I-CSF-1 to the solubilized postnuclear fraction indicated the existence of two distinct classes of binding site (apparent Kds 0.15 nM and 10 nM). A rapid assay was developed for the high-affinity sites, which were shown to be associated with the CSF-1 receptor. The function of the low-affinity sites, which have not been demonstrated on intact cells or cell membranes and which are 13 times more abundant than the high-affinity sites, is unknown. The solubilized high-affinity receptor-CSF-1 complex was stable on storage at 0 degrees C and -70 degrees C but dissociated at 37 degrees C. Dissociation also occurred at 0 degrees C in buffers of low pH (4.0) or high ionic strength (0.7 M NaCl).


Pancreas transplantation.
The treatment of patients with diabetes mellitus (DM) presents many challenges to care providers and represents a major proportion of healthcare expenditure worldwide. Successful pancreas transplantation provides durable glycemic control and improves survival for patients with diabetes. Progress in the field has mainly been based on large single center studies and the cumulative analyses of registry data from the United Network for Organ Sharing (UNOS) and the International Pancreas Transplant Registry. This review focuses on the outcomes of pancreas transplantation for patients with and without end stage renal disease. It describes the current state of pancreas transplantation, gaps in knowledge, and future studies needed to enable more patients to benefit from this treatment. A common theme that emerges is the need for multicenter randomized trials in pancreas transplantation to define clearly the efficacy, risks, and long term benefits.


Dynamical mechanisms underlying contrast gain control in single neurons.
Recent neurophysiological experiments have revealed that the linear and nonlinear kernels of the transfer function in sensory neurons are not static. Rather, they are adaptive to the contrast or the variance of time-varying input stimuli, exhibiting a contrast gain control phenomenon. We investigated the underlying biophysical causes of this phenomenon by simulating and analyzing the leaky integrate-and-fire and the Hodgkin-Huxley neuronal models. Our findings indicate that contrast gain control may result from the synergistic cooperation of the nonlinear dynamics of spike generation and the statistical properties of the stimuli. The resulting statistics-dependent stimulus threshold is shown to be a key factor underlying the adaptation of frequency tuning and amplitude gain of a neuron's transfer function in different stimulus environments.


Alteration of developmental and pathological retinal angiogenesis in angptl4-deficient mice.
Proper vessel maturation, remodeling of endothelial junctions, and recruitment of perivascular cells is crucial for establishing and maintaining vessel functions. In proliferative retinopathies, hypoxia-induced angiogenesis is associated with disruption of the vascular barrier, edema, and vision loss. Therefore, identifying factors that regulate vascular maturation is critical to target pathological angiogenesis. Given the conflicting role of angiopoietin-like-4 (ANGPTL4) reported in the current literature using gain of function systems both in vitro and in vivo, the goal of this study was to characterize angiogenesis, focusing on perinatal retinal vascularization and pathological circumstances in angpl4-deficient mice. We report altered organization of endothelial junctions and pericyte coverage, both leading to impaired angiogenesis and increased vascular leakage that were eventually caught up, suggesting a delay in vessel maturation. In a model of oxygen-induced retinopathy, pathological neovascularization, which results from tissue hypoxia, was also strongly inhibited in angptl4-deficient mice. This study therefore shows that ANGPTL4 tunes endothelial cell junction organization and pericyte coverage and controls vascular permeability and angiogenesis, both during development and in pathological conditions.


Two new leptospiral serovars in the Hebdomadis serogroup isolated from Zimbabwe cattle.
Four strains belonging to the genus Leptospira serogroup Hebdomadis were isolated from Zimbabwe cattle at slaughter. These isolates were subjected to cross-agglutinin absorption tests and to restriction fragment length polymorphism and pulsed-field gel electrophoresis analyses of their genomic DNAs. One of these strains represents a new serovar, for which the name mhou is proposed; strain SBF 40 is the reference strain of this serovar. The other three strains belong to a second new serovar, for which the name marondera is proposed; the reference strain of this serovar is strain SBF 5. The three strains of serovar marondera could be differentiated by their restriction fragment polymorphism and pulsed-field gel electrophoretic patterns.


Strength training's chronic effects on muscle architecture parameters of different arm sites.
Strength training generates alterations in muscle geometry, which can be monitored by imaging techniques as, for example, the ultrasound (US) technique. There is no consensus about the homogeneity of hypertrophy in different muscle sites. Therefore, the purpose of this study was to compare the muscle thickness (MT) and pennation angle (PA) in 3 different sites (50, 60, and 70% of arm length) of the biceps brachii and triceps brachii after 12 weeks of strength training. Forty-nine healthy untrained men were divided into 2 groups: Training Group ([TG, n = 40] 29.90 ± 1.72 years; 79.53 ± 11.84 kg; 173 ± 0.6 cm) and Control Group (n = 9 25.89 ± 3.59 years; 73.96 ± 9.86 kg; 171 ± 6 cm). The TG underwent a strength training program during 12 weeks, which included exercises such as a free-weight bench press, machine lat pull-down, triceps extension in lat pull-down, and standing free-weight biceps curl with a straight bar. A US apparatus was used to measure the PA and MT at the 3 sites. The maximal voluntary isometric contraction (MVC) test was conducted for each muscle group. After 12 weeks of training, a significant difference was observed between MT in biceps brachii, with an improvement of 12% in the proximal site, whereas the distal site increased by only 4.7% (p < 0.05). For the long head of the triceps brachii, the MT and PA at the 3 sites presented significant increases, but no significant variation was observed among them, probably because of the pennated-fiber arrangement. The MVC increased significantly for both muscle groups. The results indicated that the strength training program was efficient in promoting hypertrophy in both muscles, but with dissimilar responses of the pennated and fusiform muscle architecture at different arm sites.


Microtubule dynamics depart from the wormlike chain model.
Thermal shape fluctuations of grafted microtubules were studied using high resolution particle tracking of attached fluorescent beads. First mode relaxation times were extracted from the mean square displacement in the transverse coordinate. For microtubules shorter than approximately 10 microm, the relaxation times were found to follow an L2 dependence instead of L4 as expected from the standard wormlike chain model. This length dependence is shown to result from a complex length dependence of the bending stiffness which can be understood as a result of the molecular architecture of microtubules. For microtubules shorter than approximately 5 microm, high drag coefficients indicate contributions from internal friction to the fluctuation dynamics.


Modulation of the regulatory activity of bacterial two-component systems by SlyA.
Activation of the transcriptional regulator SlyA by the PhoP/PhoQ two-component system controls intracellular expression of numerous factors influencing Salmonella virulence. By dissecting the SlyA regulon using stable isotope labeling with amino acids in cell culture analysis, we found that SlyA enhances overall transcription of PhoP-activated loci. This amplification of cellular responses to Mg2+ occurs when SlyA binds to the phoPQ promoter thereby activating phoP autoregulation via a positive feedback mechanism. SlyA footprints a DNA region located one helical turn upstream of the PhoP box, which overlaps the H-NS-binding motif required for signal-dependent phoP repression in high Mg2+ conditions. Therefore, binding of SlyA likely antagonizes H-NS and facilitates the interaction of PhoP to its own promoter, subsequently activating the phoPQ operon. Establishment of this regulatory circuit allows SlyA to exert its effect on the PhoP/PhoQ system specifically in Salmonella, which may confer an additional transcriptional regulation. Thus, our results provide a molecular mechanism that determines SlyA-dependent activation of PhoP-regulated genes in modulating Salmonella virulence. Evidence from this study also suggests a function of SlyA as a mediator in signal transduction from the PhoP/PhoQ system to other bacterial two-component systems in Salmonella.


Breeding system and demography shape population genetic structure across ecological and climatic zones in the African freshwater snail, Bulinus forskalii (Gastropoda, Pulmonata), intermediate host for schistosomes.
The role of breeding system and population bottlenecks in shaping the distribution of neutral genetic variation among populations inhabiting patchily distributed, ephemeral water bodies was examined for the hermaphroditic freshwater snail Bulinus forskalii, intermediate host for the medically important trematode Schistosoma guineensis. Levels of genetic variation at 11 microsatellite loci were assessed for 600 individuals sampled from 19 populations that span three ecological and climatic zones (ecozones) in Cameroon, West Africa. Significant heterozygote deficiencies and linkage disequilibria indicated very high selfing rates in these populations. Despite this and the large genetic differentiation detected between populations, high levels of genetic variation were harboured within these populations. The high level of gene flow inferred from assignment tests may be responsible for this pattern. Indeed, metapopulation dynamics, including high levels of gene flow as well as extinction/contraction and recolonization events, are invoked to account for the observed population structuring, which was not a consequence of isolation-by-distance. Because B. forskalii populations inhabiting the northern, Sahelian area are subject to more pronounced annual cycles of drought and flood than the southern equatorial ones, they were expected to be subject to population bottlenecks of increased frequency and severity and, therefore, show reduced genetic variability and elevated population differentiation. Contrary to predictions, the populations inhabiting the most northerly ecozone exhibited higher genetic diversity and lower genetic differentiation than those in the most southerly one, suggesting that elevated gene flow in this region is counteracting genetic drift.


The use of multiple serial transverse enteroplasty (STEP) procedures for the management of intestinal atresia and short bowel syndrome.
The serial transverse enteroplasty procedure (STEP) was introduced as a bowel-lengthening procedure to reduce complications related to short bowel syndrome (SBS). Although some have described it as a useful adjunct to the Bianci procedure, others have acknowledged it as a primary procedure. We present a case of jejunal atresia in which two STEP procedures were performed 7 months apart to increase small bowel length. A 1-day-old, term girl presented with a known bowel obstruction diagnosed in utero. A laparotomy revealed a Type IIIb jejunal atresia with no remaining small bowel or cecum. A STEP procedure with an end jejunostomy and ascending colon mucous fistula lengthened the small bowel from 35 to 50 cm. A repeat procedure 7 months later lengthened it to 89 cm. The STEP procedure results in slower intestinal transit time and increases enterocytes contact with oral intake. We performed it during our initial exploration to increase small bowel size by 30 per cent. A repeat procedure 7 months later increased length to 89 cm. The use of multiple, staged STEP procedures avoided the need for bowel transplantation and long-term total parenteral nutrition dependence, demonstrating its effectiveness as a primary procedure for the surgical management of SBS.


Estimating under-reporting of energy intake in dietary surveys using an individualised method.
Under-reporting (UR) of energy intake (EI) by self-reported dietary methods is well-documented but the methods used to estimate UR in population-based studies commonly assume a sedentary lifestyle. We compared estimated UR using individualised estimates of energy requirements with a population cut-off based on minimum energy needs. UR was estimated for 1551 adults aged 19-64 years enrolled in the National Diet and Nutrition Survey. Physical activity diaries and 7 d weighed dietary records were completed concurrently. Mean daily EI (kJ/d) was calculated from the dietary records. Reported physical activity was used to assign each subject's activity level, and then to calculate estimated energy requirements (EER) from published equations. UR was calculated both as EER - EI with an adjustment for daily EER and EI variation, and also by a population method. By the individual method UR was approximately 27 % of energy needs in men and 29 % in women, with 75 % of men and 77 % of women classified as under-reporters; by the population method 80 and 88 % were classified as under-reporters respectively. When subjects who reported their eating being affected by dieting or illness during dietary recording were excluded, UR was 25 % of energy needs in both sexes. UR was higher in overweight and obese men and women compared with their lean counterparts (P < 0.001). UR of EI must be considered in dietary surveys. The EER method allows UR to be quantified and takes into account an individual's activity level. Measures of physical activity and questions to identify under-eating during dietary recording may help to evaluate secular trends in UR.


Windowed Fourier transform for fringe pattern analysis.
Fringe patterns in optical metrology systems need to be demodulated to yield the desired parameters. Time-frequency analysis is a useful concept for fringe demodulation, and a windowed Fourier transform is chosen for the determination of phase and phase derivative. Two approaches are developed: the first is based on the concept of filtering the fringe patterns, and the second is based on the best match between the fringe pattern and computer-generated windowed exponential elements. I focus on the extraction of phase and phase derivatives from either phase-shifted fringe patterns or a single carrier fringe pattern. Principles as well as examples are given to show the effectiveness of the proposed methods.


Evaluation of a Streamlined Oncologist-Led BRCA Mutation Testing and Counseling Model for Patients With Ovarian Cancer.
Purpose There is a growing demand for BRCA1/ 2 mutation ( BRCAm) testing in patients with ovarian cancer; however, the limited number of genetic counselors presents a potential barrier. To facilitate more widespread BRCAm testing in ovarian cancer, pretest counseling by the oncology team could shorten testing turnaround times and ease the pressure on genetic counselors. Patients and Methods The prospective, observational Evaluating a Streamlined Onco-genetic BRCA Testing and Counseling Model Among Patients With Ovarian Cancer (ENGAGE) study evaluated a streamlined, oncologist-led BRCAm testing pathway. The analysis population comprised 700 patients with ovarian cancer at 26 sites in the United States, Italy, and Spain. The primary objectives were to assess turnaround time and, using questionnaires, to evaluate stakeholder satisfaction (patients, oncologists, and geneticists or genetic counselors) with the oncologist-led BRCAm testing pathway. Results The median overall turnaround time was 9.1 weeks (range, 0.9 to 37.1 weeks), with median turnaround times in the United States, Italy, and Spain of 4.1 weeks (range, 0.9 to 37.1 weeks), 20.4 weeks (range, 2.9 to 35.4 weeks), and 12.0 weeks (range, 2.0 to 36.7 weeks), respectively. Patient satisfaction with the oncologist-led BRCAm testing pathway was high, with > 99% of patients expressing satisfaction with pre- and post- BRCAm test counseling. Oncologist satisfaction with the BRCAm testing pathway was also high, with > 80% agreeing that the process for performing BRCAm testing worked well and that counseling patients on BRCAm testing was an efficient use of their time. Oncologists expressed higher levels of satisfaction with the BRCAm testing pathway than did geneticists or genetic counselors. Conclusion The results of the ENGAGE study demonstrate that an oncologist-led BRCAm testing process is feasible in ovarian cancer. Development of local BRCAm testing guidelines similar to the one used in this study could allow faster treatment decisions and better use of resources in the management of patients with ovarian cancer.


Hepatitis G virus infection in patients transplanted for cryptogenic cirrhosis: red flag or red herring?
The significance of hepatitis G (HGV) infection in liver transplant recipients is not known. We set out to determine the pre-orthotopic liver transplantation (OLT) prevalence, the pre- and postoperative viral titers of HGV, and the allograft histology in patients infected with HGV who underwent OLT for cryptogenic cirrhosis. HGV RNA was measured using a research-based branched DNA assay. The assay used a target-specific probe set that was based on the 5'-untranslated region of the HGV genome. Allograft histology was assessed with protocol liver biopsies in all patients who survived longer than 6 months. The preoperative prevalence of HGV infection in recipients transplanted for cryptogenic cirrhosis was 26%. Thirty-seven percent (12 of 33) of recipients who had serum available in the first postoperative month had HGV infection. Mean HGV RNA levels were 9.8 (+/-4.2) (viral molecular equivalents/ml x 10[6]) before OLT and 37.5 (+/-10.7) at 1 year after OLT. In 4 of the 11 cryptogenic recipients in whom HGV RNA was detectable in the first postoperative month, HGV RNA fell to undetectable levels at the most recent follow-up (mean 70 months). Of the five cryptogenic recipients who continue to have measurable HGV RNA, three have unexplained hepatitis histologically. These findings suggest the following: 1) The prevalence of HGV infection in patients undergoing OLT for cryptogenic cirrhosis is about 25%. 2) In recipients persistently infected with HGV, mean HGV RNA titers increase after OLT. 3) HGV RNA becomes undetectable in about one third of recipients who had detectable HGV RNA in the first month after OLT. 4) Hepatitis of uncertain etiology occurs in 60% (3 of 5) of persistently HGV-infected cryptogenic recipients.


Attenuation of airway obliteration by ciprofloxacin in experimental posttransplant bronchiolitis obliterans.
Ciprofloxacin is widely used to treat respiratory tract infections. Like other fluoroquinolones, ciprofloxacin has immunomodulatory effects; however, it is unknown whether these effects are beneficial in the setting of lung transplantation. We investigated potential immunomodulatory effects of ciprofloxacin in a model of posttransplant bronchiolitis obliterans. The heterotopic tracheal transplantation model in rats was used. Three groups received ciprofloxacin and underwent different immunosuppressive regimens of cyclosporine A, that is, no immunosuppression, insufficient immunosuppression, or low-dose immunosuppression. Three groups underwent the same immunosuppressive regimen but had no ciprofloxacin treatment. Tracheas were harvested after 21 days and examined with respect to histology and expression of selected cytokines. The allografts of animals treated with ciprofloxacin showed less airway obliteration compared with allografts of untreated animals. When combined with low-dose immunosuppression ciprofloxacin showed beneficial effects in preventing airway obliteration and rejection of the respiratory epithelium. Cytokine gene expression of the allografts treated with ciprofloxacin was higher with respect to transforming growth factor-beta and equal with respect to tumor necrosis factor-alpha and interferon-gamma compared with controls. When applied in combination with cyclosporine A, ciprofloxacin lowered the expression of transforming growth factor-beta and tumor necrosis factor-alpha and increased interferon-gamma expression. Ciprofloxacin attenuates airway rejection after tracheal transplantation. Genetic expression of mediators that are known to play an important role in mediating rejection in this model supports an immunomodulatory and antifibrotic role of ciprofloxacin. These findings suggest that further clinical studies are needed to investigate whether ciprofloxacin in addition to its bactericidal effect might be beneficial in the treatment of human posttransplant bronchiolitis obliterans.


Sodium and potassium urinary excretion and dietary intake: a cross-sectional analysis in adolescents.
Hypertension is the leading cause for heart disease and stroke, for mortality and morbidity worldwide, and a high sodium-to-potassium intake ratio is considered a stronger risk factor for hypertension than sodium alone. This study aims to evaluate sodium and potassium urinary excretion, and assess the food sources of these nutrients in a sample of Portuguese adolescents. A cross-sectional study with a sample of 250 Portuguese adolescents. Sodium and potassium excretion were measured by one 24-h urinary collection, and the coefficient of creatinine was used to validate completeness of urine collections. Dietary sources of sodium and potassium were assessed using a 24-h dietary recall. Valid urine collections were provided by 200 adolescents (118 girls) with a median age of 14.0 in both sexes (p=0.295). Regarding sodium, the mean urinary excretion was 3,725 mg/day in boys and 3,062 mg/day in girls (p<0.01), and 9.8% of boys and 22% of girls met the World Health Organization (WHO) recommendations for sodium intake. Concerning potassium, the mean urinary excretion was 2,237 mg/day in boys and 1,904 mg/day in girls (p<0.01), and 6.1% of boys and 1.7% of girls met the WHO recommendations for potassium intake. Major dietary sources for sodium intake were cereal and cereal products (41%), meat products (16%), and milk and milk products (11%); and for potassium intake, main sources were milk and milk products (21%), meat products (17%), and vegetables (15%). Adolescents had a high-sodium and low-potassium diet, well above the WHO recommendations. Health promotion interventions are needed in order to decrease sodium and increase potassium intake.


Self-etch and etch-and-rinse adhesive systems in clinical dentistry.
Current adhesive systems follow either an "etch-and-rinse" or "self-etch" approach, which differ in how they interact with natural tooth structures. Etch-and-rinse systems comprise phosphoric acid to pretreat the dental hard tissues before rinsing and subsequent application of an adhesive. Self-etch adhesives contain acidic monomers, which etch and prime the tooth simultaneously. Etch-and-rinse adhesives are offered as two- or three-step systems, depending on whether primer and bonding are separate or combined in a single bottle. Similarly, self-etch adhesives are available as one- or two-step systems. Both etch-and-rinse and self-etch systems form a hybrid layer as a result of resins impregnating the porous enamel or dentin. Despite current trends toward fewer and simpler clinical application steps, one-step dentin bonding systems exhibit bonding agent lower bond strengths and seem less predictable than multi-step etch-and-rinse and self-etch systems. The varying evidence available today suggests that the choice between etch-and-rinse and self-etch systems is often a matter of personal preference. In general, however, phosphoric acid creates a more pronounced and retentive etching pattern in enamel. Therefore, etch-and-rinse bonding systems are often preferred for indirect restorations and when large areas of enamel are still present. Conversely, self-etch adhesives provide superior and more predictable bond strength to dentin and are, consequently, recommended for direct composite resin restorations, especially when predominantly supported by dentin.


SHORT-COMMUNICATION Evaluation of perfused bovine udder for gene expression studies in dairy cows.
Intramammary infections are one of the main causes of productivity loss in dairy cows. To better understand the immune system response and to avoid the use of live animals, we validated the use of isolated bovine udder as an ex situ model. Six mammary glands were collected from cows ready for culling. Three udders were perfused with Tyrode's solution and three were not-perfused. During six hours, we collected perfusate samples for biochemical analysis. We also collected alveolar and teat canal tissue to evaluate gene expression. The biochemical parameters indicated that the perfused udders remained viable for the entire period of the experiment. A real-time polymerase chain reaction showed an increase in 18S rRNA gene expression in the alveolar tissue at 3 and 4 h after perfusion. There was also an increase in the Ubiquitin gene in the teat canal from not-perfused udders at 1, 3, and 4 h after slaughter. In general, gene expression was stable during the experiment. Our results indicated that the isolated perfused bovine udder model is appropriate for genetic studies, opening a new perspective in animal experimentation methods.


Isostructural series of nine-coordinate chiral lanthanide complexes based on triazacyclononane.
Nonadentate ligands based on triazacyclononane incorporating pyridyl-2-phosphinate groups form an isostructural series of complexes with Ln ions in the solid state and in solution. The Ln ion is effectively shielded from the solvent environment. Crystal structures reveal a rigid C(3)-symmetric tricapped trigonal-prismatic coordination geometry that is maintained in solution for the methyl and phenylphosphinate series, as shown by multinuclear NMR analysis. Variable-temperature measurements of the field dependence of the water proton relaxivity in gadolinium complexes indicate that these systems exclude solvent from the primary coordination environment and minimize the second sphere of solvation. The electronic relaxation time for the gadolinium methylphosphinate complex has been estimated to be 550 (±150) ps by EPR and NMR methods, compared to values of around 0.30-0.05 ps for the terbium-ytterbium series, deduced by analyzing the field dependence (4.7-16.5 T) of the (31)P NMR longitudinal relaxation times. Values are compared with analogous azacarboxylate ligand complexes, supporting a key role for donor atom polarizability in determining the electronic relaxation. Spectral emission behavior in solution of samarium, europium, terbium, and dysprosium complexes is compared, and the resolved RRR-Λ and SSS-Δ complexes show strong circularly polarized luminescence. The molecular quadratic hyperpolarizability 〈β(HLS)〉 has been measured in solution using hyper-Raleigh light-scattering methods, for the whole series of lanthanide complexes of one ligand. The values of 〈β(HLS)〉 reach a maximum around the center of the series and are not simply dependent on the number of f electrons, suggesting a dominant contribution from the octupolar rather than the dipolar term.


A monoclonal antibody to Bacillus anthracis protective antigen defines a neutralizing epitope in domain 1.
Antibody (Ab) responses to Bacillus anthracis toxins are protective, but relatively few protective monoclonal antibodies (MAbs) have been reported. Protective antigen (PA) is essential for the action of B. anthracis lethal toxin (LeTx) and edema toxin. In this study, we generated two MAbs to PA, MAbs 7.5G and 10F4. These MAbs did not compete for binding to PA, consistent with specificities for different epitopes. The MAbs were tested for their ability to protect a monolayer of cultured macrophages against toxin-mediated cytotoxicity. MAb 7.5G, the most-neutralizing MAb, bound to domain 1 of PA and reduced LeTx toxicity in BALB/c mice. Remarkably, MAb 7.5G provided protection without blocking the binding of PA or lethal factor or the formation of the PA heptamer complex. However, MAb 7.5G slowed the proteolytic digestion of PA by furin in vitro, suggesting a potential mechanism for Ab-mediated protection. These observations indicate that some Abs to domain 1 can contribute to host protection.


Modified-Release and Conventional Glucocorticoids and Diurnal Androgen Excretion in Congenital Adrenal Hyperplasia.
The classic androgen synthesis pathway proceeds via dehydroepiandrosterone, androstenedione, and testosterone to 5α-dihydrotestosterone. However, 5α-dihydrotestosterone synthesis can also be achieved by an alternative pathway originating from 17α-hydroxyprogesterone (17OHP), which accumulates in congenital adrenal hyperplasia (CAH). Similarly, recent work has highlighted androstenedione-derived 11-oxygenated 19-carbon steroids as active androgens, and in CAH, androstenedione is generated directly from 17OHP. The exact contribution of alternative pathway activity to androgen excess in CAH and its response to glucocorticoid (GC) therapy is unknown. We sought to quantify classic and alternative pathway-mediated androgen synthesis in CAH, their diurnal variation, and their response to conventional GC therapy and modified-release hydrocortisone. We used urinary steroid metabolome profiling by gas chromatography-mass spectrometry for 24-hour steroid excretion analysis, studying the impact of conventional GCs (hydrocortisone, prednisolone, and dexamethasone) in 55 adults with CAH and 60 controls. We studied diurnal variation in steroid excretion by comparing 8-hourly collections (23:00-7:00, 7:00-15:00, and 15:00-23:00) in 16 patients with CAH taking conventional GCs and during 6 months of treatment with modified-release hydrocortisone, Chronocort. Patients with CAH taking conventional GCs showed low excretion of classic pathway androgen metabolites but excess excretion of the alternative pathway signature metabolites 3α,5α-17-hydroxypregnanolone and 11β-hydroxyandrosterone. Chronocort reduced 17OHP and alternative pathway metabolite excretion to near-normal levels more consistently than other GC preparations. Alternative pathway-mediated androgen synthesis significantly contributes to androgen excess in CAH. Chronocort therapy appears superior to conventional GC therapy in controlling androgen synthesis via alternative pathways through attenuation of their major substrate, 17OHP.


Learning temporal context shapes prestimulus alpha oscillations and improves visual discrimination performance.
Time is an inseparable component of every physical event that we perceive, yet it is not clear how the brain processes time or how the neuronal representation of time affects our perception of events. Here we asked subjects to perform a visual discrimination task while we changed the temporal context in which the stimuli were presented. We collected electroencephalography (EEG) signals in two temporal contexts. In predictable blocks stimuli were presented after a constant delay relative to a visual cue, and in unpredictable blocks stimuli were presented after variable delays relative to the visual cue. Four subsecond delays of 83, 150, 400, and 800 ms were used in the predictable and unpredictable blocks. We observed that predictability modulated the power of prestimulus alpha oscillations in the parieto-occipital sites: alpha power increased in the 300-ms window before stimulus onset in the predictable blocks compared with the unpredictable blocks. This modulation only occurred in the longest delay period, 800 ms, in which predictability also improved the behavioral performance of the subjects. Moreover, learning the temporal context shaped the prestimulus alpha power: modulation of prestimulus alpha power grew during the predictable block and correlated with performance enhancement. These results suggest that the brain is able to learn the subsecond temporal context of stimuli and use this to enhance sensory processing. Furthermore, the neural correlate of this temporal prediction is reflected in the alpha oscillations.NEW & NOTEWORTHY It is not well understood how the uncertainty in the timing of an external event affects its processing, particularly at subsecond scales. Here we demonstrate how a predictable timing scheme improves visual processing. We found that learning the predictable scheme gradually shaped the prestimulus alpha power. These findings indicate that the human brain is able to extract implicit subsecond patterns in the temporal context of events.


Crystallographic Structure Determination of Both [5,6]- and [6,6]-Isomers of Lithium-Ion-Containing Diphenylmethano[60]fullerene.
Organic functionalization of lithium-ion-containing [60]fullerene, Li+@C60, was performed by using diphenyl(diazo)methane as a stable, readily available diazo compound to obtain lithium-ion-containing [5,6]- and [6,6]-diphenylmethano[60]fullerenes, Li+@C61Ph2. The bis(trifluoromethanesulfonyl)imide (TFSI) salts of [5,6]- and [6,6]-Li+@C61Ph2 were successfully separated by using a cation exchange column with eluent containing LiTFSI. Improved separation protocol and high crystallinity of ionic components in less polar solvents enabled separate crystallization of each isomer. Both [5,6]-open and [6,6]-closed structures of Li+@C61Ph2 were determined by synchrotron radiation X-ray crystallography. Elucidating the [5,6]-open methano[60]fullerene (fulleroid) structure will contribute to materials research on fulleroids.


Achieving universal health coverage in South Africa through a district health system approach: conflicting ideologies of health care provision.
Universal Health Coverage (UHC) has emerged as a major goal for health care delivery in the post-2015 development agenda. It is viewed as a solution to health care needs in low and middle countries with growing enthusiasm at both national and global levels. Throughout the world, however, the paths of countries to UHC have differed. South Africa is currently reforming its health system with UHC through developing a national health insurance (NHI) program. This will be practically achieved through a decentralized approach, the district health system, the main vehicle for delivering services since democracy. We utilize a review of relevant documents, conducted between September 2014 and December 2015 of district health systems (DHS) and UHC and their ideological underpinnings, to explore the opportunities and challenges, of the district health system in achieving UHC in South Africa. Review of data from the NHI pilot districts suggests that as South Africa embarks on reforms toward UHC, there is a need for a minimal universal coverage and emphasis on district particularity and positive discrimination so as to bridge health inequities. The disparities across districts in relation to health profiles/demographics, health delivery performance, management of health institutions or district management capacity, income levels/socio-economic status and social determinants of health, compliance with quality standards and above all the burden of disease can only be minimised through positive discrimination by paying more attention to underserved and disadavantaged communities. We conclude that in South Africa the DHS is pivotal to health reform and UHC may be best achieved through minimal universal coverage with positive discrimination to ensure disparities across districts in relation to disease burden, human resources, financing and investment, administration and management capacity, service readiness and availability and the health access inequalities are consciously implicated. Yet ideological and practical issues make its achievement problematic.


Improving postpartum screening after diabetes in pregnancy: Results of a pilot study in remote Australia.
The postpartum period is a critical time to improve health outcomes for Aboriginal women, particularly for those who have chronic conditions. To assess enhanced support methods (for women following diabetes in pregnancy (DIP)) to improve completion rates of recommended postpartum health checks. Fifty-three Aboriginal women in the Northern Territory (NT) were contacted in the postpartum period to encourage medical check-ups. Messages were delivered through phone (call or text messages) or other methods (Facebook or email). The primary outcome was postpartum blood glucose testing (oral glucose tolerance testing (OGTT), random or fasting glucose and HbA1c). Establishing contact with women was difficult. Of 137 messages sent to 52 women, 22 responded (42%). Phone was the most common contact method with successful contact made from 16 of 119 (13%) attempts. Rates of postpartum OGTT completion were higher in the group successfully contacted (32% vs 7%). However, for any postpartum glucose testing (including OGTT and HbA1c) rates were 25 of 42 (60%) and neither success in making contact nor the contact method was associated with higher rates. The small sample size limits our conclusions; however, results highlight that engaging remote women postpartum is difficult. While rates of postpartum OGTT completion differed according to successful contacts, rates of any postpartum blood glucose testing did not. Further research is needed to explore feasible intervention methods to improve postpartum screening after a pregnancy complicated by diabetes.


High levels of acute phase proteins and soluble 70 kDa heat shock proteins are independent and additive risk factors for mortality in colorectal cancer.
Recently, we reported that high soluble Hsp70 (sHsp70) level was a significant predictor of mortality during an almost 3-year-long follow-up period in patients with colorectal cancer. This association was the strongest in the group of <70-year-old female patients as well as in those who were in a less advanced stage of the disease at baseline. According to these observations, measurement of the serum level of sHsp70 is a useful, stage-independent prognostic marker in colorectal cancer, especially in patients without distant metastasis. Since many literature data indicated that measurement of C-reactive protein (CRP) and other acute phase proteins (APPs) may also be suitable for predicting the mortality of patients with colorectal cancer, it seemed reasonable to study whether the effect of sHsp70 and other APPs are related or independent. In order to answer this question, we measured the concentrations of CRP as well as of other complement-related APPs (C1 inhibitor, C3, and C9) along with that of the MASP-2 complement component in the sera of 175 patients with colorectal cancer and known levels of sHsp70, which have been used in our previous study. High (above median) levels of CRP, C1 esterase inhibitor (C1-INH), and sHsp70 were found to be independently associated with poor patient survival, whereas no such association was observed with the other proteins tested. According to the adjusted Cox proportional hazards analysis, the additive effect of high sHsp70, CRP, and C1-INH levels on the survival of patients exceeded that of high sHsp70 alone, with a hazard ratio (HR) of 2.83 (1.13-70.9). In some subgroups of patients, such as in females [HR 4.80 (1.07-21.60)] or in ≤70-year-old patients [HR 11.53 (2.78-47.70)], even greater differences were obtained. These findings indicate that the clinical mortality-prediction value of combined measurements of sHsp70, CRP, and C1-INH with inexpensive methods can be very high, especially in specific subgroups of patients with colorectal cancer.


Predominance of fetal type DJH joining in young children with B precursor lymphoblastic leukemia as evidence for an in utero transforming event.
The presence of N sequences in the complementarity determining region 3 (CDR3) of the rearranged immunoglobulin H chain is developmentally regulated: N regions are generally present in the DJH joinings of adult B cells but are often absent in fetal B cells. Analysis of the CDR3 in 61 B precursor acute lymphoblastic leukemias indicated that 87.5% of the leukemias obtained from children < or = 3 yr old lacked N regions at the DJH junction. In contrast, in children > 3 yr old, only 11.1% of the leukemias lacked N regions at this junction, a frequency similar to what we have observed in B cells from children and adults. These findings suggest that the majority of leukemias presenting within the first 3 yr of age arise from an in utero transforming event.


Terminator still moving forward: expanding roles for Rho factor.
Rho factor is a molecular motor that translocates along nascent RNA and acts on the transcription elongation complex to promote termination. Besides contributing to transcriptional punctuation of the bacterial genome, Rho can act intragenically under conditions that perturb coupling of translation and transcription. Recent advances have shed new light onto several aspects of Rho function, including the translocation mechanism, the avoidance of potential conflicts between DNA replication and transcription, suppression of pervasive antisense transcription and recruitment in riboswitch and small RNA-dependent regulation. Altogether, these findings further highlight the relevance of Rho factor, both as a multi-task housekeeper and gene regulator.


Resolution upgrade toward 6-bit optical quantization using power-to-wavelength conversion for photonic analog-to-digital conversion.
We demonstrate a resolution upgrade toward 6 bit optical quantization using a power-to-wavelength conversion without an increment of system parallelism. Expansion of a full-scale input range is employed in conjunction with reduction of a quantization step size with keeping a sampling-rate transparency characteristic over several 100 sGS/s. The effective number of bits is estimated to 5.74 bit, and the integral nonlinearity error and differential nonlinearity error are estimated to less than 1 least significant bit.


Obesity Takes Its Toll on Visceral Pain: High-Fat Diet Induces Toll-Like Receptor 4-Dependent Visceral Hypersensitivity.
Exposure to high-fat diet induces both, peripheral and central alterations in TLR4 expression. Moreover, functional TLR4 is required for the development of high-fat diet-induced obesity. Recently, central alterations in TLR4 expression have been associated with the modulation of visceral pain. However, it remains unknown whether there is a functional interaction between the role of TLR4 in diet-induced obesity and in visceral pain. In the present study we investigated the impact of long-term exposure to high-fat diet on visceral pain perception and on the levels of TLR4 and Cd11b (a microglial cell marker) protein expression in the prefrontal cortex (PFC) and hippocampus. Peripheral alterations in TLR4 were assessed following the stimulation of spleenocytes with the TLR4-agonist LPS. Finally, we evaluated the effect of blocking TLR4 on visceral nociception, by administering TAK-242, a selective TLR4-antagonist. Our results demonstrated that exposure to high-fat diet induced visceral hypersensitivity. In parallel, enhanced TLR4 expression and microglia activation were found in brain areas related to visceral pain, the PFC and the hippocampus. Likewise, peripheral TLR4 activity was increased following long-term exposure to high-fat diet, resulting in an increased level of pro-inflammatory cytokines. Finally, TLR4 blockage counteracted the hyperalgesic phenotype present in mice fed on high-fat diet. Our data reveal a role for TLR4 in visceral pain modulation in a model of diet-induced obesity, and point to TLR4 as a potential therapeutic target for the development of drugs to treat visceral hypersensitivity present in pathologies associated to fat diet consumption.


Regional inhibition of cholinesterase in free-ranging western pond turtles (Emys marmorata) occupying California mountain streams.
The present study investigated the potential effects of cholinesterase (ChE)-inhibiting pesticides on western pond turtles (Emys marmorata) occupying streams in two regions of California, USA. The southern region was suspected of having increased exposure to atmospheric deposition of contaminants originating from Central Valley agriculture. The northern region represented reference ChE activities because this area was located outside of the prominent wind patterns that deposit pesticides into the southern region. Total ChE activity was measured in plasma from a total of 81 turtles from both regions. Cholinesterase activity of turtles was significantly depressed by 31% (p = 0.005) in the southern region after accounting for additional sources of variation in ChE activity. Male turtles had significantly increased ChE activity compared with females (p = 0.054). Cloaca temperature, length, mass, handling time, body condition, and lymph presence were not significant predictors of turtle ChE activity. In the southern region, 6.3% of the turtles were below the diagnostic threshold of two standard deviations less than the reference site mean ChE activity. Another diagnostic threshold determined that 75% of the turtles from the southern region had ChE activities depressed by 20% of the reference mean. The decrease in ChE activity in the southern region suggests sublethal effects of pesticide exposure, potentially altering neurotransmission, which can result in various deleterious behaviors.


The effect of high-altitude on human skeletal muscle energetics: P-MRS results from the Caudwell Xtreme Everest expedition.
Many disease states are associated with regional or systemic hypoxia. The study of healthy individuals exposed to high-altitude hypoxia offers a way to explore hypoxic adaptation without the confounding effects of disease and therapeutic interventions. Using (31)P magnetic resonance spectroscopy and imaging, we investigated skeletal muscle energetics and morphology after exposure to hypobaric hypoxia in seven altitude-naïve subjects (trekkers) and seven experienced climbers. The trekkers ascended to 5300 m while the climbers ascended above 7950 m. Before the study, climbers had better mitochondrial function (evidenced by shorter phosphocreatine recovery halftime) than trekkers: 16+/-1 vs. 22+/-2 s (mean +/- SE, p<0.01). Climbers had higher resting [Pi] than trekkers before the expedition and resting [Pi] was raised across both groups on their return (PRE: 2.6+/-0.2 vs. POST: 3.0+/-0.2 mM, p<0.05). There was significant muscle atrophy post-CXE (PRE: 4.7+/-0.2 vs. POST: 4.5+/-0.2 cm(2), p<0.05), yet exercising metabolites were unchanged. These results suggest that, in response to high altitude hypoxia, skeletal muscle function is maintained in humans, despite significant atrophy.


Alexithymia, Prior Trauma, Alcohol Use, and Sexual Aggression Perpetration: A Cross-Cultural Comparison of a Moderated Mediation Model.
In this study, we examined the relationships between prior trauma, alexithymia, and sexual aggression perpetration among 610 U.S. college students and 107 college students from the Philippines utilizing a cross-sectional retrospective design. We tested a moderated mediation model with alexithymia as the mediator between prior trauma and sexual aggression perpetration, and alcohol use as a moderator of the alexithymia-sexual aggression link. Moreover, given that cultural norms may influence these relationships, we also examined the structural invariance of the proposed moderated mediation model. Path analyses and multiple group analysis were used to examine the moderated mediation model, and examine model differences between samples. We did not find evidence for alexithymia as a mediator, but there was a significant interaction between alcohol use severity and alexithymia on sexual aggression perpetration across both samples. Alexithymia is a key variable in understanding the alcohol use-sexual aggression perpetration relationship. Clinical implications and recommendations for future research are also discussed.


Transcriptional mechanisms underlying sensitization of peripheral sensory neurons by granulocyte-/granulocyte-macrophage colony stimulating factors.
Cancer-associated pain is a major cause of poor quality of life in cancer patients and is frequently resistant to conventional therapy. Recent studies indicate that some hematopoietic growth factors, namely granulocyte macrophage colony stimulating factor (GMCSF) and granulocyte colony stimulating factor (GCSF), are abundantly released in the tumor microenvironment and play a key role in regulating tumor-nerve interactions and tumor-associated pain by activating receptors on dorsal root ganglion (DRG) neurons. Moreover, these hematopoietic factors have been highly implicated in postsurgical pain, inflammatory pain and osteoarthritic pain. However, the molecular mechanisms via which G-/GMCSF bring about nociceptive sensitization and elicit pain are not known. In order to elucidate G-/GMCSF mediated transcriptional changes in the sensory neurons, we performed a comprehensive, genome-wide analysis of changes in the transcriptome of DRG neurons brought about by exposure to GMCSF or GCSF. We present complete information on regulated genes and validated profiling analyses and report novel regulatory networks and interaction maps revealed by detailed bioinformatics analyses. Amongst these, we validate calpain 2, matrix metalloproteinase 9 (MMP9) and a RhoGTPase Rac1 as well as Tumor necrosis factor alpha (TNFα) as transcriptional targets of G-/GMCSF and demonstrate the importance of MMP9 and Rac1 in GMCSF-induced nociceptor sensitization. With integrative approach of bioinformatics, in vivo pharmacology and behavioral analyses, our results not only indicate that transcriptional control by G-/GMCSF signaling regulates a variety of established pain modulators, but also uncover a large number of novel targets, paving the way for translational analyses in the context of pain disorders.


Two members of an HNF1 homeoprotein family are expressed in human liver.
HNF1 is a transcriptional activator, required for the liver-specific expression of a variety of genes, that binds to DNA as a dimer via the most diverged homeodomain known so far. We were interested to examine whether HNF1 is a unique homeoprotein example or whether it is the prototype of a new subfamily of homeodomain containing proteins. In this work we describe the isolation of a cDNA clone from a human liver library encoding a protein, highly homologous to HNF1 in three regions, including the homeo- and dimerization domains. We show that this protein can heterodimerize with human HNF1 in vitro. Sequence comparison of our clone with a rat variant HNF1 (vHNF1) clone, isolated in parallel in our laboratory from the dedifferentiated H5 hepatoma cell line, identified our cDNA as human vHNF1. vHNF1 is a nuclear protein recognizing the same binding site as HNF1 and previously thought to occur only in dedifferentiated hepatoma cells that fail to express most liver specific genes. Nevertheless, we show by Northern blot analysis that vHNF1 transcripts are present in differentiated human HepG2 hepatoma cells as well as in rat liver and that this transcript level is 10-20 fold lower than that of HNF1. We assigned the vHNF-1 gene to human chromosome 17 and murine chromosome 11. These chromosomal localizations differ from that of the HNF-1 gene indicating that both genes are not clustered on the genome.


Polyunsaturated fatty acids, DNA repair single nucleotide polymorphisms and colorectal cancer in the Singapore Chinese Health Study.
Animal and in vitrostudies support a role for polyunsaturated fatty acids (PUFAs) in colon carcinogenesis; however, the epidemiological evidence is inconclusive. Recently, we investigated their role within the Singapore Chinese Health Study, a population-based cohort of Singapore Chinese men and women. We reported that a high intake of marine n-3 PUFAs was associated with an increased risk of colorectal cancer (CRC). Oxidation of PUFAs incorporated into cell membranes generates lipid hydroperoxides, which can be mutagenic. In this report, we investigated whether single nucleotide polymorphisms (SNPs) in DNA repair genes modified the effect of PUFAs on CRC risk using a nested case-control study within the Singapore Chinese Health Study. We genotyped 1,181 controls and 311 cases (180 colon and 131 rectal cancer) for SNPs in the XRCC1 (Arg194Trp, Arg399Gln), OGG1 (Ser326Cys), PARP (Val762Ala, Lys940Arg), and XPD (Asp312Asn, Lys751Gln) genes. We observed that the PARP Val762Ala SNP modified the association between marine n-3 PUFA and rectal cancer risk, with no evidence of interaction among colon cancer (heterogeneity test p=0.003). Our results suggest a positive association between high intake of marine n-3 PUFA and rectal cancer risk among carriers of at least one PARP codon 762 Ala allele (odds ratio=1.7, 95% confidence interval=1.1-2.7).


Generalized B-spline subdivision-surface wavelets for geometry compression.
We present a new construction of lifted biorthogonal wavelets on surfaces of arbitrary two-manifold topology for compression and multiresolution representation. Our method combines three approaches: subdivision surfaces of arbitrary topology, B-spline wavelets, and the lifting scheme for biorthogonal wavelet construction. The simple building blocks of our wavelet transform are local lifting operations performed on polygonal meshes with subdivision hierarchy. Starting with a coarse, irregular polyhedral base mesh, our transform creates a subdivision hierarchy of meshes converging to a smooth limit surface. At every subdivision level, geometric detail can be expanded from wavelet coefficients and added to the surface. We present wavelet constructions for bilinear, bicubic, and biquintic B-Spline subdivision. While the bilinear and bicubic constructions perform well in numerical experiments, the biquintic construction turns out to be unstable. For lossless compression, our transform can be computed in integer arithmetic, mapping integer coordinates of control points to integer wavelet coefficients. Our approach provides a highly efficient and progressive representation for complex geometries of arbitrary topology.


Influence of ions on aqueous acid-base reactions.
We study the effects of bromide salts on the rate and mechanism of the aqueous proton/deuteron-transfer reaction between the photoacid 8-hydroxy-1,3,6-pyrenetrisulfonic acid (HPTS) and the base acetate. The proton/deuteron release is triggered by exciting HPTS with 400 nm femtosecond laser pulses. Probing the electronic and vibrational resonances of the photoacid, the conjugate photobase, the hydrated proton/deuteron and the accepting base with femtosecond visible and mid-infrared pulses monitors the proton transfer. Two reaction channels are identified: 1) direct long-range proton transfer over hydrogen-bonded water bridges that connect the acid and base and 2) acid dissociation to produce fully solvated protons followed by proton scavenging from solution by acetate. We observe that the addition of salt affects the long-range reaction pathway, and reduces both the rate at which protons are released to solution by HPTS and the rate at which solvated protons are scavenged from solution by acetate. We study the dependence of these effects on the nature and concentration of the dissolved salt.


Biocompatibility of Four Common Orthopedic Biomaterials Following a High-Salt Diet: An In Vivo Study.
Nowadays, salt consumption appears to be drastically above the recommended level in industrialized countries. The health consequences of this overconsumption are heavy since high-salt intake induces cardiovascular disease, kidney dysfunction, and stroke. Moreover, harmful interaction may also occur with orthopaedic devices because overconsumption of salt reinforces the corrosive aspect of biological tissues and favors bone resorption process. In the present study, we aimed to assess the in vivo effect of three weeks of a high-salt diet, associated (or not) with two weeks of the neuro-myoelectrostimulation (NMES) rehabilitation program on the biocompatibility of four biomaterials used in the manufacture of arthroplasty implants. Thus, two non-metallic (PEEK and Al₂O₃) and two metallic (Ti6Al4V and CrCo) compounds were implanted in the rat tibial crest, and the implant-to-bone adhesion and cell viability of two surrounded muscles, the Flexor Digitorum (FD) and Tibialis Anterior (TA), were assessed at the end of the experiment. Results indicated lower adhesion strength for the PEEK implant compared to other biomaterials. An effect of NMES and a high-salt diet was only identified for Al₂O₃ and Ti6Al4V implants, respectively. Moreover, compared to a normal diet, a high-salt diet induced a higher number of dead cells on both muscles for all biomaterials, which was further increased for PEEK, Al₂O₃, and CrCo materials with NMES application. Finally, except for Ti6Al4V, NMES induced a higher number of dead cells in the directly stimulated muscle (FD) compared to the indirectly stimulated one (TA). This in vivo experiment highlights the potential harmful effect of a high-salt diet for people who have undergone arthroplasty, and a rehabilitation program based on NMES.


Committee Opinion No.703: Asymptomatic Microscopic Hematuria in Women.
Asymptomatic microscopic hematuria is an important clinical sign of urinary tract malignancy. Asymptomatic microscopic hematuria has been variably defined over the years. In addition, the evidence primarily is based on data from male patients. However, whether the patient is a man or a woman influences the differential diagnosis of asymptomatic microscopic hematuria, and the risk of urinary tract malignancy (bladder, ureter, and kidney) is significantly less in women than in men. Among women, being older than 60 years, having a history of smoking, and having gross hematuria are the strongest predictors of urologic cancer. In low-risk, never-smoking women younger than 50 years without gross hematuria and with fewer than 25 red blood cells per high-power field, the risk of urinary tract malignancy is less than or equal to 0.5%. Furthermore, the evaluation may result in more harm than benefit and is unlikely to be cost effective. Thus, data support changing current hematuria recommendations in this low-risk group. The American College of Obstetricians and Gynecologists and the American Urogynecologic Society encourage organizations producing future guidelines on the evaluation of microscopic hematuria to perform sex-specific analysis of the data and produce practical sex-specific recommendations. In the meantime, the American College of Obstetricians and Gynecologists and the American Urogynecologic Society recommend that asymptomatic, low-risk, never-smoking women aged 35-50 years undergo evaluation only if they have more than 25 red blood cells per high-power field.


Chitosan/gelatin hydrogel as immunoisolative matrix for injectable bioartificial pancreas.
The feasibility of using chitosan/gelatin hydrogel as immunoisolative matrix to provide an additional protection to the microencapsulated islet cells was demonstrated in this study. We hope that the use of hydrogel can extend the functional longevity of microencapsulated islet cells during xenotransplantation. Chitosan/gelatin solution with glycerol 2-phosphate disodium salt hydrate was prepared and utilized as a cell carrier. The biocompatibility of the chitosan/gelatin hydrogel was first established by using a mouse insulinoma cell line, NIT-1. Insulinoma cells were encapsulated in agarose as microspheres and then macroencapsulated in chitosan/gelatin hydrogel. In vitro cell activity, material-mediated cytotoxicity, cytokine-mediated cytotoxicity assay, and insulin secreting profiles of insulinoma/agarose microspheres macroencapsulated in chitosan/gelatin hydrogel were analyzed. For in vivo study, insulinoma/agarose microspheres with chitosan/gelatin solution was applied as an injectable bioartificial pancreas (BAPs). Insulinoma/agarose microspheres suspended in phosphate-buffered saline or in chitosan/gelatin solution was injected into the subcutaneous layer of diabetic rats. Non-fasting blood glucose (NFBG) concentration of rat was measured perioperatively. After pre-determined intervals, the chitosan/gelatin hydrogel containing insulinoma/agarose microspheres was retrieved for histologic examinations. Insulinoma/agarose microspheres macroencapsulated in hydrogel revealed functional activity and secreted insulin continually for 60 days in vitro. Chitosan/gelatin hydrogel was not cytotoxic to islet cells, and in contrast, the hydrogel showed cytoprotective effects against cytokine-mediated cytotoxicity. The NFBG of diabetic rats transplanted with free insulinoma/agarose microspheres was decreased to euglycemia but restored to hyperglycemia in 15 days. Contrarily, the NFBG of rats transplanted with insulinoma/agarose microspheres with hydrogel remained euglycemic for 42 days. Histologic sections revealed that the fibrous tissue envelopment and the infiltrated immune-related cells contributed to the dysfunction of BAPs. This study indicates that using chitosan/gelatin hydrogel as a cell carrier is feasible and can provide an additional protection for the microencapsulated islet cells during xenotransplantation.


Anomalous transport and nonlinear reactions in spiny dendrites.
We present a mesoscopic description of the anomalous transport and reactions of particles in spiny dendrites. As a starting point we use two-state Markovian model with the transition probabilities depending on residence time variable. The main assumption is that the longer a particle survives inside spine, the smaller becomes the transition probability from spine to dendrite. We extend a linear model presented in Fedotov [Phys. Rev. Lett. 101, 218102 (2008)] and derive the nonlinear Master equations for the average densities of particles inside spines and parent dendrite by eliminating residence time variable. We show that the flux of particles between spines and parent dendrite is not local in time and space. In particular the average flux of particles from a population of spines through spines necks into parent dendrite depends on chemical reactions in spines. This memory effect means that one cannot separate the exchange flux of particles and the chemical reactions inside spines. This phenomenon does not exist in the Markovian case. The flux of particles from dendrite to spines is found to depend on the transport process inside dendrite. We show that if the particles inside a dendrite have constant velocity, the mean particle's position <x(t)> increases as t(μ) with μ<1 (anomalous advection). We derive a fractional advection-diffusion equation for the total density of particles.


The roots of bioinformatics in theoretical biology.
From the late 1980s onward, the term "bioinformatics" mostly has been used to refer to computational methods for comparative analysis of genome data. However, the term was originally more widely defined as the study of informatic processes in biotic systems. In this essay, I will trace this early history (from a personal point of view) and I will argue that the original meaning of the term is re-emerging.


Complications of ankle arthroscopy utilizing a contemporary noninvasive distraction technique.
The reported complication rates of ankle arthroscopy, as well as the types of complications, refer to cases performed fifteen to twenty years ago and include a mixture of invasive and noninvasive distraction methods. As ankle arthroscopy has matured with a move away from invasive distraction and the introduction of improved instruments, it is unclear whether the previously reported complications are reflective of contemporary ankle arthroscopy. We sought to evaluate the types and rates of complications that are associated with contemporary ankle arthroscopy that utilizes noninvasive ankle distraction and a padded foot strap. We retrospectively reviewed 294 consecutive arthroscopies. All patients underwent the same intraoperative noninvasive distraction protocol. Demographic data, diagnoses, duration of follow-up data, and procedure-related complications were identified, recorded, and analyzed. All 294 cases were eligible for review. There were twenty complications, resulting in an overall complication rate of 6.8%. There were four non-neurologic complications, including one case of deep venous thrombosis, one case of prolonged portal drainage, and two cases of superficial infection. There were sixteen neurologic complications; six were related to the anterolateral portal, and eight patients had dysesthesias involving the dorsal part of the midfoot. When analyzed as a group, patients who were receiving Workers' Compensation had a higher complication rate than patients who were not receiving Workers' Compensation (21% versus 4.7%, respectively). Contemporary ankle arthroscopy performed with use of a noninvasive distraction technique is a safe procedure. Our observed complication rate of 6.8% is lower than that reported in a previous benchmark study, although our patients who were receiving Workers' Compensation had a significantly higher risk of having nerve symptoms. An important benefit of the use of contemporary ankle arthroscopy, in which small-joint arthroscopic instruments and noninvasive distraction techniques are used, is a marked reduction in the rate of non-neurologic complications.


Rapid through-container detection of fake spirits and methanol quantification with handheld Raman spectroscopy.
The spirits drinks industry is of significant global economic importance and a major employer worldwide, and the ability to ensure product authenticity and maintain consumer confidence in these high-value products is absolutely essential. Spirit drinks counterfeiting is a worldwide problem, with counterfeiting and adulteration of spirit drinks taking many forms, such as substitution, stretching with lower-grade products, or creation of counterfeits with industrial, surrogate, or locally produced alcohols. Methanol for example, which has been used as a substitute alcohol for ethanol, has a high toxicity in humans. The counterfeiting of spirit drinks is consequently one of the few leading reported types of food fraud which can be directly and unequivocally linked to food safety and health concerns. Here, for the first time, we use handheld Raman spectroscopy with excitation in the near IR (1064 nm) for the through-container differentiation of multiple spirit drinks, detection of multiple chemical markers of counterfeit alcohol, and for the quantification of methanol. We established the limits of detection (LOD) of methanol in the analysed samples from four different spirit types (between 0.23-0.39%), which were considerably lower than a quoted maximum tolerable concentration (MTC) of 2% (v/v) methanol for humans in a 40% alcohol by volume (ABV) spirit drink, and even lower than the general EU limit for naturally occurring methanol in fruit spirits of 0.5% v/v (10 g methanol per L ethanol). We believe that Raman spectroscopy has considerable practicable potential for the rapid in situ through-container detection of counterfeit spirits drinks, as well as for the analysis and protection of other beverages and liquid samples.


Genic interaction causing embryonic mortality in the rat: epistasis between the Tal and grc genes.
The autosomal dominant mutant gene, tail anomaly lethal (Tal), of the rat is lethal when homozygous but affects tail morphology (kinks and reduced length) and body weight when heterozygous. There is no apparent sex effect on the expression of Tal. It is incompletely penetrant; has variable expressivity, which is influenced partly by its genetic background; and is not linked to the major histocompatibility complex (MHC). The heterozygous Tal gene and the homozygous grc genes, which are linked to the MHC and affect body size and fertility, interact to cause intrauterine death at a time between implantation (five to seven days post-fertilization) and 15 days of gestation. This interaction shifts the time of death from the immediate postnatal period when the homozygous grc genes act to the time during gestation when the homozygous Tal gene would cause death. This description of lethal epistatic interaction in the rat appears to be the first report of this phenomenon in mammals.


NTP Toxicology and Carcinogenesis Studies of Chlorinated Trisodium Phosphate (CAS No. 56802-99-4)* in B6C3F1 Mice (Gavage Studies).
Two-year toxicology and carcinogenesis studies of chlorinated trisodium phosphate, an inclusion complex of trisodium phosphate and sodium hypochlorite used in various cleaning compounds, were conducted by administering 0, 500, or 1,000 mg/kg (dose volume: 10 ml/kg) of the chemical in water by gavage, 5 days per week for 103 weeks, to groups of 50 male and 50 female B6C3F1 mice. Groups of mice receiving 250 mg/kg were included in these studies but were removed after 6 months because of a lack of toxicity in the 500 and 1,000 mg/kg groups. Two-year studies were begun in male and female F344/N rats at doses of 0, 500, 1,000, or 2,000 mg/kg of chlorinated trisodium phosphate in water by gavage (10 ml/kg). The 2,000 mg/kg groups were killed at 15 weeks because of poor survival, and the other groups were killed at 35 weeks because of toxicity in the 1,000 mg/kg group. The doses selected for the 2-year studies were based on the general lack of adverse effects seen in the 14-day and 13-week studies in which rats received 0-1,000 mg/kg and mice received 0-2,000 mg/kg by gavage in water. No compound-related histopathologic effects were observed in the 14-day or the 13-week studies in mice. In the 2-year studies, survival and mean body weights of dosed and vehicle control male mice groups were comparable (survival-- vehicle control, 39/50; low dose, 35/50; high dose, 32/50). Survival of the dosed female mice was lower than that of the vehicle controls (30/50; 16/50; 21/50), although at week 80 survival of female mice was 42/50, 39/50, and 36/50. The mean body weights of the high dose female mice were lower than those of the vehicle control mice, primarily after week 32; final body weights were 11% lower in the high dose group compared with that in the vehicle controls. The lower survival and mean body weights of the dosed female mice may have been due to a greater incidence of uterine/ovarian infections in these mice rather than to a direct toxic effect of chlorinated trisodium phosphate. Nine of 20 vehicle control, 20/34 low dose, and 21/29 high dose female mice that died before the end of the studies had such infections. This reduced survival decreased the sensitivity of the study of female mice for detecting the presence or absence of carcinogenic effects. At no site was the incidence of neoplasms considered to be related to the administration of chlorinated trisodium phosphate. Minimal necrosis and fatty changes were observed in the livers of male mice. Kidneys in male mice were characterized by small, multifocal areas of mineralization, primarily in the cortex but not at the corticomedullary junction or in the tubes of the medulla. Neither effect was considered compound related. Five different types of ovarian neoplasms were found in six dosed female mice; because these lesions were from tissues of different embryonic origin, they were considered unrelated to administration of chlorinated trisodium phosphate. Chlorinated trisodium phosphate was weakly mutagenic in strain TA1535 of Salmonella typhimurium in the presence of Aroclor 1254-induced male Sprague-Dawley rat or male Syrian hamster liver S9. This compound was not mutagenic in strains TA97, TA98, or TA100. An audit of the experimental data was conducted for these 2-year studies of chlorinated trisodium phosphate. No data discrepancies were found that influenced the final interpretation of these experiments. Under the conditions of these 2-year gavage studies, there was no evidence of carcinogenicity for either male or female B6C3F1 mice given chlorinated trisodium phosphate by gavage in water for 103 weeks at doses of 500 or 1,000 mg. Survival of dosed female mice was 78% and 72% after 80 weeks and 32% and 42% at the termination of the study. The studies in male and female F344/N rats were considered to be inadequate studies of carcinogenicity because the experiments were terminated at 35 weeks due to poor survival. Synonym: sodium hypochlorite phosphate


Anesthetic techniques for fetal surgery: effects of maternal anesthesia on intraoperative fetal outcomes in a sheep model.
Use of high-dose inhalational anesthesia during open fetal surgery may induce maternal-fetal hemodynamic instability and fetal myocardial depression. The authors' preliminary human retrospective study demonstrated less fetal bradycardia and left ventricular systolic dysfunction with lower dose desflurane supplemented with propofol and remifentanil IV anesthesia (SIVA). In this animal study, the authors compare maternal-fetal effects of high-dose desflurane anesthesia (HD-DES) and SIVA. Of 26 instrumented midgestational ewes, data from 11 animals exposed to both SIVA and HD-DES in random sequences and six animals exposed to HD-DES while maternal normotension was maintained were analyzed. Maternal electroencephalography was used to guide comparable depths of anesthesia in both techniques. Hemodynamic parameters, blood gas, and fetal cardiac function from echocardiography were recorded. Compared with SIVA, HD-DES resulted in significant maternal hypotension (mean arterial pressure difference, 19.53 mmHg; 95% CI, 17.6-21.4; P < 0.0001), fetal acidosis (pH 7.11 vs. 7.24 at 150 min, P < 0.001), and decreased uterine blood flow. In the HD-DES group with maternal normotension, uterine blood flow still declined and fetal acidosis persisted, with no statistically significant difference from the group exposed to HD-DES that had maternal hypotension. There was no statistically significant difference in fetal cardiac function. In sheep, SIVA affects maternal hemodynamics less and provides better fetal acid/base status than high-dose desflurane. Fetal echocardiography did not reflect myocardial dysfunction in this model.


Solitary pancreatic lymph node metastasis from carcinoma of the breast: case report.
We report the first case of isolated pancreatic lymph node recurrence in a locally advanced breast cancer patient. A 41-year old woman underwent radical mastectomy according to Madden and removal of axillary lymph nodes for multicentric infiltrating ductal carcinoma pathologically staged as pT2N2M0. After six years from primary diagnosis, and four years from the diagnosis of lung recurrence, she developed an isolated metastatic lesion to pancreatic lymph node. After surgical excision of metastasis, hormone therapy with Exemestane was begun. At 16 months of follow-up, the patient appears free of disease. Because metastatization to visceral organ carries a very unfavorable prognosis, we think that the clinical significance of the elevation of CA 15.3 serum levels in the early detection of recurrence and in monitoring metastatic disease during follow-up, should be not underestimated.


Clinical importance of Stenotrophomonas maltophilia nosocomial pneumonia due to its high mortality in hemodialysis patients.
Patients undergoing hemodialysis are immunocompromised and can suffer from pneumonia with various pathogens in nosocomial conditions. We investigated the fundamental information on the characteristics of hemodialysis inpatients and nosocomial pneumonia. We surveyed 1803 hemodialysis patients admitted to our university hospital between 2001 and 2007. The mean patient age was 64.8 years and the average period of hospitalization was 28.1 days, which was considerably longer than the average stay in our hospital (14.2 days). Patients were admitted to many different departments and for various reasons. We isolated 391 microorganisms from the sputum of 120 pneumonia patients undergoing hemodialysis, including Candida albicans, methicillin-resistant Staphylococcus aureus, and Staphylococcus epidermidis, which were the leading three isolates. From these 120 patients, a total of 199 pathogens were identified as being responsible for the pneumonia. Multi-drug resistant Stenotrophomonas maltophilia was found to be susceptible to a new fluoroquinolone, but is resistant to older generation quinolones. Out of the 120 patients with pneumonia, 12 out of 18 patients infected with S. maltophilia died, indicating the highest fatality rate for this pathogen. In this survey, we found that hemodialysis patients were hospitalized for long periods, and for various reasons in many departments. They suffered from nosocomial pneumonia caused by multi-drug resistant pathogens, including S. maltophilia. For pneumonia due to S. maltophilia, new generation fluoroquinolones can be the treatment of choice, although S. maltophilia-related pneumonia should be treated very carefully because of its high fatality rate.


An efficient solution to systems of multivariate polynomial using expression trees.
In recent years, several quite successful attempts have been made to solve systems of polynomial constraints, using geometric design tools, exploiting the availability of subdivision-based solvers [7], [11], [12], [15]. This broad range of methods includes both binary domain subdivision as well as the projected polyhedron method of Sherbrooke and Patrikalakis [15]. A prime obstacle in using subdivision solvers is their scalability. When the given constraint is represented as a tensor product of all its independent variables, it grows exponentially in size as a function of the number of variables. In this work, we show that for many applications, especially geometric ones, the exponential complexity of the constraints can be reduced to a polynomial by representing the underlying structure of the problem in the form of expression trees that represent the constraints. We demonstrate the applicability and scalability of this representation and compare its performance to that of tensor product constraint representation through several examples.


Predictive value of multislice computed tomography variables of atherosclerosis for ischemia on stress-rest single-photon emission computed tomography.
previous studies have shown that the presence of stenosis alone on multislice computed tomography (MSCT) has a limited positive predictive value for the presence of ischemia on myocardial perfusion imaging (MPI). The purpose of this study was to assess which variables of atherosclerosis on MSCT angiography are related to ischemia on MPI. both MSCT and MPI were performed in 514 patients. On MSCT, the calcium score, degree of stenosis (≥ 50% and ≥ 70% stenosis), and plaque extent and location were determined. Plaque composition was classified as noncalcified, mixed, or calcified. Ischemia was defined as a summed difference score (≥ 2 on a per-patient basis. Ischemia was observed in 137 patients (27%). On a per-patient basis, multivariate analysis showed that the degree of stenosis (presence of (≥ 70% stenosis, odds ratio=3.5), plaque extent and composition (mixed plaques (≥3, odds ratio=1.7; calcified plaques ≥ 3, odds ratio=2.0), and location (atherosclerotic disease in the left main coronary artery and/or proximal left anterior descending coronary artery, odds ratio=1.6) were independent predictors for ischemia on MPI. In addition, MSCT variables of atherosclerosis, such as plaque extent, composition, and location, had significant incremental value for the prediction of ischemia over the presence of ≥70% stenosis. in addition to the degree of stenosis, MSCT variables of atherosclerosis describing plaque extent, composition, and location are predictive of the presence of ischemia on MPI.


Order of removal of conventional and nonconventional introns from nuclear transcripts of Euglena gracilis.
Nuclear genes of euglenids and marine diplonemids harbor atypical, nonconventional introns which are not observed in the genomes of other eukaryotes. Nonconventional introns do not have the conserved borders characteristic for spliceosomal introns or the sequence complementary to U1 snRNA at the 5' end. They form a stable secondary structure bringing together both exon/intron junctions, nevertheless, this conformation does not resemble the form of self-splicing or tRNA introns. In the genes studied so far, frequent nonconventional introns insertions at new positions have been observed, whereas conventional introns have been either found at the conserved positions, or simply lost. In this work, we examined the order of intron removal from Euglena gracilis transcripts of the tubA and gapC genes, which contain two types of introns: nonconventional and spliceosomal. The relative order of intron excision was compared for pairs of introns belonging to different types. Furthermore, intermediate products of splicing were analyzed using the PacBio Next Generation Sequencing system. The analysis led to the main conclusion that nonconventional introns are removed in a rapid way but later than spliceosomal introns. Moreover, the observed accumulation of transcripts with conventional introns removed and nonconventional present may suggest the existence of a time gap between the two types of splicing.


Activation of the ATR-mediated DNA damage response by the HIV-1 viral protein R.
DNA damage is a universal inducer of cell cycle arrest at the G2 phase. Infection by the human immunodeficiency virus type 1 (HIV-1) also blocks cellular proliferation at the G2 phase. The HIV-1 accessory gene vpr encodes a conserved 96-amino acid protein (Vpr) that is necessary and sufficient for the HIV-1-induced block of cellular proliferation. In the present study, we examined a recently identified DNA damage-signaling protein, the ATM- and Rad3-related protein, ATR, for its potential role in the induction of G2 arrest by Vpr. We show that inhibition of ATR by pharmacological inhibitors, by expression of the dominant-negative form of ATR, or by RNA interference inhibits Vpr-induced cell cycle arrest. As with DNA damage, activation of ATR by Vpr results in phosphorylation of Chk1. This study provides conclusive evidence of activation of the ATR-initiated DNA damage-signaling pathway by a viral gene product. These observations are important toward understanding how HIV infection promotes cell cycle disruption, cell death, and ultimately, CD4+ lymphocyte depletion.


Large-scale recent expansion of European patrilineages shown by population resequencing.
The proportion of Europeans descending from Neolithic farmers ∼ 10 thousand years ago (KYA) or Palaeolithic hunter-gatherers has been much debated. The male-specific region of the Y chromosome (MSY) has been widely applied to this question, but unbiased estimates of diversity and time depth have been lacking. Here we show that European patrilineages underwent a recent continent-wide expansion. Resequencing of 3.7 Mb of MSY DNA in 334 males, comprising 17 European and Middle Eastern populations, defines a phylogeny containing 5,996 single-nucleotide polymorphisms. Dating indicates that three major lineages (I1, R1a and R1b), accounting for 64% of our sample, have very recent coalescent times, ranging between 3.5 and 7.3 KYA. A continuous swathe of 13/17 populations share similar histories featuring a demographic expansion starting ∼ 2.1-4.2 KYA. Our results are compatible with ancient MSY DNA data, and contrast with data on mitochondrial DNA, indicating a widespread male-specific phenomenon that focuses interest on the social structure of Bronze Age Europe.


From gene identification to gene therapy.
Inner ear disease due to hair cell loss is common, and no restorative treatments for the balance and hearing impairment are currently available. To develop clinical means for enhancing protection and regeneration in the inner ear, it is necessary to understand the molecular basis for hereditary and acquired deafness and vestibular disorders. One approach is to identify and characterize genes that regulate protection or repair in other systems. For that purpose, we have used the differential display assay and compared gene expression between normal and acoustically traumatized inner ears of chicks. Several chick cDNAs that were identified are considered as candidates for roles in the reparative process that follows trauma in the basilar papilla. The mammalian vestibular epithelium has a limited regenerative capability. To identify genes that may participate in the regenerative response, we have used gene arrays profiling, comparing normal to drug-traumatized vestibular epithelia. We identified several genes that are differentially expressed in traumatized vestibular epithelium, including several insulin-like growth factor-I binding proteins. To use this molecular knowledge for enhancing protection and repair in the organ of Corti, it is necessary to overexpress the genes of choice in the inner ear. Using viral-mediated gene transfer, we overexpressed transgenic glial cell line-derived neurotrophic factor and demonstrated a robust protective effect against acoustic and ototoxic inner ear trauma. Future identification of the genes that are important for protection and regeneration, along with improved gene transfer technology, will allow the use of gene therapy for treating hereditary and environmental inner ear disease.


Comparison of bispectral index values and depth of sedation during deep sedation using sevoflurane anesthesia in healthy children versus children with cerebral palsy.
Patients with cerebral palsy (CP) are at significant risk by means of periodontal disease and tooth decay. Pharmacological techniques that require intensive care such as sedation and general anesthesia are generally used for dental treatment of this patient group. The purpose of this retrospective study was to compare bispectral index (BIS) values and depth of sedation recorded during deep sedation protocols performed for healthy children and children with CP in the Department of Pediatric Dentistry, Faculty of Dentistry, Gazi University. The measurements of BIS and hemodynamic variables recorded during treatment of 26 healthy children and 26 children with CP between 3 and 10 years of age who were treated under sevoflurane and 50% N2O-50% O2 mixture deep sedation method were investigated retrospectively. The mean BIS values in the CP group was statistically lower at all the time points when compared with the control group (P < 0.001). While there was no statistically significant difference between groups in terms of duration of treatment (P = 0.657), the median recovery time in the CP group was significantly longer than that recorded in the control group (P < 0.001). Significant correlation was found between modified Ramsay Sedation Scale (mRSS) scores and BIS levels at 5th, 10th, and 20th min in the control group (P < 0.001). Similar correlation was found in the CP group at 15th and 20th min. We concluded that it is necessary to consider the dosage and effect mechanisms of drugs used in children with CP to prevent overuse of anesthetics and emergence of anesthesia-related complications.


Alternans and spiral breakup in a human ventricular tissue model.
Ventricular fibrillation (VF) is one of the main causes of death in the Western world. According to one hypothesis, the chaotic excitation dynamics during VF are the result of dynamical instabilities in action potential duration (APD) the occurrence of which requires that the slope of the APD restitution curve exceeds 1. Other factors such as electrotonic coupling and cardiac memory also determine whether these instabilities can develop. In this paper we study the conditions for alternans and spiral breakup in human cardiac tissue. Therefore, we develop a new version of our human ventricular cell model, which is based on recent experimental measurements of human APD restitution and includes a more extensive description of intracellular calcium dynamics. We apply this model to study the conditions for electrical instability in single cells, for reentrant waves in a ring of cells, and for reentry in two-dimensional sheets of ventricular tissue. We show that an important determinant for the onset of instability is the recovery dynamics of the fast sodium current. Slower sodium current recovery leads to longer periods of spiral wave rotation and more gradual conduction velocity restitution, both of which suppress restitution-mediated instability. As a result, maximum restitution slopes considerably exceeding 1 (up to 1.5) may be necessary for electrical instability to occur. Although slopes necessary for the onset of instabilities found in our study exceed 1, they are within the range of experimentally measured slopes. Therefore, we conclude that steep APD restitution-mediated instability is a potential mechanism for VF in the human heart.


An increase in telomere sister chromatid exchange in murine embryonic stem cells possessing critically shortened telomeres.
Telomerase deficiency leads to a progressive loss of telomeric DNA that eventually triggers cell apoptosis in human primary cells during prolonged growth in culture. Rare survivors can maintain telomere length through either activation of telomerase or recombination-based telomere lengthening, and thus proliferate indefinitely. We have explored the possibility that telomeres may be maintained through telomere sister chromatid exchange (T-SCE) in murine telomere reverse transcriptase-deficient (mTert-/-) splenocytes and ES cells. Because telomerase deficiency leads to gradual loss of telomeric DNA in mTert-/- splenocytes and ES cells and eventually to chromosomes with telomere signal-free ends (SFEs), we examined these cell types for evidence of sister chromatid exchange at telomeres, and observed an increase in T-SCEs only in a subset of mTert-/- splenocytes or ES cells that possessed multiple SFEs. Furthermore, T-SCEs were more often detected in ES cells than in splenocytes that harbored a similar frequency of SFEs. In mTert heterozygous (mTert+/-) ES cells or splenocytes, which are known to exhibit a decrease in average telomere length but no SFEs, no increase in T-SCE was observed. In addition to T-SCE, other genomic rearrangements (i.e., SCE) were also significantly increased in mTert-/- ES cells possessing critically short telomeres, but not in splenocytes. Our results suggest that animals and cell culture differ in their ability to carry out genomic rearrangements as a means of maintaining telomere integrity when telomeres become critically shortened.