The efficacy of zinc for treatment of chronic prostatitis.
to investigate the efficacy of zinc supplementation in chronic prostatitis treatment. present randomized clinical trial was conducted on 120 patients with diagnosis of chronic prostatitis (IIIA NIH) after preliminary evaluation and ruling out other conditions. The study group received oral zinc sulfate 220 mg daily as capsule without any other supplements. The control group received placebo. Subjects were examined for NIH-CPSI scores every 4 weeks for 12 weeks. 101 subjects completed the study. There were no statistically significant differences in scores and sub-scores of NIH-CPSI between groups before intervention. Decline in the score and sub-scores were more prominent in case group after beginning of the study; though the differences were not statistically significant. Furthermore, the differences in total score and pain score at 12 weeks follow was statistically significant (p=0.003 and p=0.02, respectively). zinc supplements may benefit in management of patients with chronic prostatitis NIH-IIIA. It can be attributable to anti-bacterial and immuno-modulatory functions of organic zinc in the body.


Epidemiological Study on Candida Species in Patients with Cancer in the Intensive Care Unit.
Although cancer survival rates have increased, serious infection complications can arise in cancer patients. Candida can occur in various tissues and has significant effects on the prognosis of patients with cancer. Thus, we conducted an epidemiological study on Candida infections in patients with cancer admitted to the intensive care unit. A retrospective study was conducted in adult patients with cancer admitted to the intensive care unit between January 1, 2013, and December 31, 2015. Candida infection status and predictive factors for mortality were examined in 634 patients. The predictive factors for mortality included the use of steroids, use of a central venous catheter or mechanical ventilator, and identification of Candida in the blood. Patients who stayed in the surgical and intensive care unit for more than 7 days had a lower risk of death than that in those with shorter days. The present study shows that invasive procedures, bloodstream infections, and the use of steroids increase the risk of mortality in Candida-infected patients with cancer. To improve the quality of life and reduce mortality, further studies are needed on the factors affecting the risk of mortality associated with Candida infection.


Unfolding IGDQ Peptides for Engineering Motogenic Interfaces.
Extracellular matrix (ECM)-mimicking surfaces are pivotal tools in understanding adherent cell physiopathology. In this sense, we have recently reported on a discrete set of ECM-mimicking SAMs, among which only those exposing IGDQ peptide-alkanethiols sustain the adhesion of MDA-MB-231 cells by triggering FAK phosphorylation and peculiarly induce the migration of individual cancer cells on the subcentimeter scale. Starting from the experimentally observed relationship among the SAM composition, organization, and biological response, a systematic computational characterization aided in pinpointing the atomistic details through which specific composition and organization achieve the desired biological responsiveness. Specifically, the solvent, number and type of peptides, and presence or absence of surface fillers were accurately considered, creating representative model SAMs simulated by means of classical molecular dynamics (MD) with a view toward unravelling the experimental evidence, revealing how the conformational and structural features of these substrates dictate the specific motogenic responses. Through complementary experimental and computational investigations, it clearly emerges that there exists a distinct and precise mutual interaction among IGDQ-peptides, the surface fillers, and Au, which controls the structural properties of the ECM-mimicking SAMs and thus their motogenic potential.


Communication: direct angle-resolved measurements of collision dynamics with electronically excited molecules: NO(A2Σ+) + Ar.
We report direct doubly differential (quantum state and angle-resolved) scattering measurements involving short-lived electronically excited molecules using crossed molecular beams. In our experiment, supersonic beams of nitric oxide and argon atoms collide at 90°. In the crossing region, NO molecules are excited to the A(2)Σ(+)state by a pulsed nanosecond laser, undergo rotationally inelastic collisions with Ar atoms, and are then detected 400 ns later (approximately twice the radiative lifetime of the A(2)Σ(+)state) by 1 + 1(') multiphoton ionization via the E(2)Σ(+) state. The velocity distributions of the scattered molecules are recorded using velocity-mapped ion imaging. The resulting images provide a direct measurement of the state-to-state differential scattering cross sections. These results demonstrate that sufficient scattering events occur during the short lifetimes typical of molecular excited states (∼200 ns, in this case) to allow spectroscopically detected quantum-state-resolved measurements of products of excited-state collisions.


Cloning, characterization, and expression of a 200-kilodalton diagnostic antigen of Babesia bigemina.
Current serological tests for Babesia bigemina use semipurified merozoite antigens derived from infected erythrocytes. One of the major drawbacks of these tests is that antigen quality can vary from batch to batch. Since the quality of the antigen contributes to the sensitivity and specificity of serological tests, the use of standardized recombinant antigens should ensure consistency in assay quality. Previously, a 200-kDa merozoite antigen (p200) was identified as a candidate diagnostic antigen for use in a serological assay for the detection of B. bigemina antibodies in infected cattle. In this study, we have cloned, characterized, and expressed p200. A 3.5-kbp cDNA clone encoding p200 was isolated and shown to be almost full length, lacking approximately 300 bp at the 5' end. The predicted amino acid sequence shows that p200 consists of a long, highly charged central repeat region of an uninterrupted alpha helix, indicative of a fibrous protein. Immunoelectron microscopy localized p200 to the merozoite cytoplasm, suggesting that the antigen may be a structural protein involved in forming filament structures within the cytoskeleton. The 3.5-kbp cDNA was expressed in bacteria as a fusion protein with glutathione S-transferase (GST), but the yield was poor. To improve the yield, cDNA fragments encoding antigenic domains of p200 were expressed as fusions with GST. One of these fusion proteins, C1A-GST, is composed of a 7-kDa fragment of the p200 repeat region and contains epitopes that react strongly with sera from cattle experimentally infected with B. bigemina. Recombinant C1A-GST should permit the development of an improved enzyme-linked immunosorbent assay for the detection of antibodies against B. bigemina.


A comparison of two methods for assessing awareness of antitobacco television advertisements.
This study uses an online survey panel to compare two approaches for assessing ad awareness. The first uses a screenshot of a television ad and the second shows participants a full-length video of the ad. We randomly assigned 1034 Minnesota respondents to view a screenshot or a streaming video from two antitobacco ads. The study used one ad from ClearWay Minnesota's ITALIC! We All Pay the Price campaign, and one from the Centers for Disease Control ITALIC! Tips campaign. The key measure used to assess ad awareness was aided ad recall. Multivariate analyses of recall with cessation behaviour and attitudinal beliefs assessed the validity of these approaches. The respondents who saw the video reported significantly higher recall than those who saw the screenshot. Associations of recall with cessation behaviour and attitudinal beliefs were stronger and in the anticipated direction using the screenshot method. Over 20% of the respondents assigned to the video group could not see the ad. People who were under 45 years old, had incomes greater than $35,000 and women were reportedly less able to access the video. The methodology used to assess recall matters. Campaigns may exaggerate the successes or failures of their media campaigns, depending on the approach they employ and how they compare it to other media campaign evaluations. When incorporating streaming video, researchers should consider accessibility and report possible response bias. Researchers should fully define the measures they use, specify any viewing accessibility issues, and make ad comparisons only when using comparable methods.


Defensive coping styles in chronic fatigue syndrome.
The cognitive-behavioral model of chronic fatigue syndrome (CFS) proposes that rigidly held beliefs act to defend individuals against low self-esteem. This study is the first to investigate the prevalence of a potential mechanism, the Defensive High Anxious coping style, among individuals with CFS. The study comprised 68 participants (24 CFS; 24 healthy volunteers; 20 chronic illness volunteers). Participants completed the Bendig short form of the Taylor Manifest Anxiety Scale (B-MAS) and the Marlowe-Crowne Social Desirability Scale (MC) in order to ascertain the distribution of participants in each group within the four coping styles defined by Weinberger et al. [J. Abnorm. Psychol. 88 (1979) 369]. A greater number of participants in the CFS group (46%) were classified as Defensive High Anxious compared to the two comparison groups [chi(2)(2)=8.84, P=.012]. This study provides support for the existence of defensive coping mechanisms as described by the cognitive-behavioral model of CFS. Furthermore, it has been suggested that this particular coping style may impinge directly on physical well being through similar mechanisms as identified in CFS, and further research linking these areas of research is warranted.


Stem cell transplantation for Huntington's diseases.
Therapeutic approaches based on stem cells have received considerable attention as potential treatments for Huntington's disease (HD), which is a fatal, inherited neurodegenerative disorder, caused by progressive loss of GABAergic medium spiny neurons (MSNs) in the striatum of the forebrain. Transplantation of stem cells or their derivatives in animal models of HD, efficiently improved functions by replacing the damaged or lost neurons. In particular, neural stem cells (NSCs) for HD treatments have been developed from various sources, such as the brain itself, the pluripotent stem cells (PSCs), and the somatic cells of the HD patients. However, the brain-derived NSCs are difficult to obtain, and the PSCs have to be differentiated into a population of the desired neuronal cells that may cause a risk of tumor formation after transplantation. In contrast, induced NSCs, derived from somatic cells as a new stem cell source for transplantation, are less likely to form tumors. Given that the stem cell transplantation strategy for treatment of HD, as a genetic disease, is to replace the dysfunctional or lost neurons, the correction of mutant genes containing the expanded CAG repeats is essential. In this review, we will describe the methods for obtaining the optimal NSCs for transplantation-based HD treatment and the differentiation conditions for the functional GABAergic MSNs as therapeutic cells. Also, we will discuss the valuable gene correction of the disease stem cells by the CRISPR/Cas9 system for HD treatment.


Pelvic osteotomies in hip dysplasia: why, when and how?
Acetabular dysplasia is a significant problem in the spectrum of developmental dysplasia of hip. In a younger child, positioning the femoral head into the acetabulum helps in reciprocal remodeling of the acetabulum and correction of dysplasia. In an older child, the remodeling potential is limited and often the acetabular dysplasia needs surgical intervention in the form of a pelvic osteotomy. Thus, pelvic osteotomy forms an integral part of surgical management of hip dysplasia. The ultimate goal of these osteotomies is to preclude or postpone the development of osteoarthritis and add more years of life to the native hip. Pelvic osteotomies play a pivotal role in normalizing hip morphology. The choice of pelvic osteotomy depends on the age of a child, the type of dysplasia and the status of the tri-radiate cartilage. Several types of re-directional and reshaping pelvic osteotomies have been described in the literature to improve the stability and restore the anatomy and biomechanics of the dysplastic hip. This article attempts to review the current indications for various pelvic osteotomies with a brief description of their techniques along with the outcomes and complications published thus far. Besides, the guidelines to choose the right pelvic osteotomy are also provided.


THE EFFECT OF 6-MERCAPTOPURINE AND AMINOPTERIN ON EXPERIMENTAL IMMUNE THYROIDITIS IN GUINEA PIGS.
The influence of 6-mercaptopurine and aminopterin was studied on immune response and immune thyroiditis in guinea pigs immunized with thyroid extract. 1. Both compounds depressed delayed hypersensitivity to thyroglobulin and immune thyroiditis. 2. Antibody formation to thyroglobulin was strongly depressed by aminopterin but not significantly influenced by 6-mercaptopurine. 3. Immune response and thyroiditis were suppressed as long as the compounds were administered; after discontinuation of treatment, immune response and thyroiditis appeared in the same time intervals as observed in control animals after initiation of immunization. 4. Treatment with 6-mercaptopurine and aminopterin 10 days after immunization lead to suppression of delayed hypersensitivity and thyroiditis. 5. Treatment with 6-mercaptopurine of animals after onset of thyroiditis lead to suppression of delayed hypersensitivity and disappearance or diminution of round cell infiltration in the thyroid. 6. The results are discussed in terms of the pathogenesis of experimental immune thyroiditis, the mode of action of these antimetabolites on this experimental immune disease, and in view of the potential value of these compounds in human diseases.


Quantitative determination of neutrophil VCS parameters by the Coulter automated hematology analyzer: new and reliable indicators for acute bacterial infection.
We studied the clinical usefulness of the VCS parameters (mean channels of cell volume, conductivity, and light scatter) in reactive neutrophils for predicting acute bacterial infection, which are obtained by the Coulter LH 750 hematology analyzer (Beckman Coulter, Fullerton, CA) during automated differential counts. Peripheral blood samples from 69 patients with positive blood cultures for bacteria and 35 control subjects were studied. We observed a significant increase in the mean channel of neutrophil volume (MNV) from septic patients compared with control subjects (156+/-13.5 vs 143+/-4.8; P<.001). The mean channel of neutrophil light scatter was decreased significantly in patients (140+/-10.1 vs 146+/-7.3; P=.002). An elevation of the MNV was associated with a higher WBC count and percentage of neutrophils and was present even in patients who did not have leukocytosis or neutrophilia. With a cutoff of 150 for the MNV, a specificity of 91% and sensitivity of 70% were achieved. As a quantitative, objective, and more sensitive parameter, we believe that the MNV has a potential to be an additional indicator for acute bacterial infection.


Identification and characterization of putative methylation targets in the MAOA locus using bioinformatic approaches.
Monoamine oxidase A (MAO A) is an enzyme that catalyzes the oxidation of neurotransmitter amines. A functional polymorphism in the human MAOA gene (high- and low-MAOA) has been associated with distinct behavioral phenotypes. To investigate directly the biological mechanism whereby this polymorphism influences brain function, we recently measured the activity of the MAO A enzyme in healthy volunteers. When found no relationship between the individual's brain MAO A level and the MAOA genotype, we postulated that there are additional regulatory mechanisms that control the MAOA expression. Given that DNA methylation is linked to the regulation of gene expression, we hypothesized that epigenetic mechanisms factor into the MAOA expression. Our underplaying assumption was that the differences in an individual's genotype play a key role in the epigenetic potential of the MAOA locus and, consequently, determine the individual's level of MAO A activity in the brain. As a first step towards experimental validation of the hypothesis, we performed a comprehensive bioinformatic analysis aiming to interrogate genomic features and attributes of the MAOA locus that might modulate its epigenetic sensitivity. Major findings of our analysis are the following: (1) the extended MAOA regulatory region contains two CpG islands (CGIs), one of which overlaps with the canonical MAOA promoter and the other is located further upstream; both CGIs exhibit sensitivity to differential methylation. (2) The uVNTR's effect on the MAOA's transcriptional activity might have epigenetic nature: this polymorphic region resides within the MAOA's CGI and itself contains CpGs, thus, the number of repeating increments effectively changes the number of methylatable cytosines in the MAOA promoter. An array of in silico analyses (the nucleosome positioning, the physical properties of the local DNA, the clustering of transcription-factor binding sites) together with experimental data on histone modifications and Pol 2 sites and data from the RefSeq mRNA library suggest that the MAOA gene might have an alternative promoter. Based on our findings, we propose a regulatory mechanism for the human MAOA according to which the MAOA expression in vivo is executed by the generation of tissue-specific transcripts initiated from the alternative promoters (both CGI-associated) where transcriptional activation of a particular promoter is under epigenetic control.


Diffusion-based model of pulse oximetry: in vitro and in vivo comparisons.
A model of pulse oximetry is developed based on the three-dimensional photon diffusion theory. To test the applicability of the model, an in vitro assay was developed. Three different scattering levels and six different relative dye concentrations were analyzed. Average percent errors of 13.9% were obtained over the full range of the study. An in vivo clinical study of two pulse oximeter probes with different spectral characteristics was compared with results estimated by the model. The model correctly predicted the changes in pulse oximeter response resulting from the wavelength changes. A x(2) test gave a probability of 20% that the model fit the data. These results demonstrated the utility of the photon diffusion theory for the modeling of tissue optics.


Identification of resected root-end dentinal cracks: a comparative study of visual magnification.
The purpose of this in vitro study was to evaluate and compare the effectiveness of visual enhancements as aids in identifying artificially created dentinal cracks in resected root ends. Fifty human maxillary central incisors were decoronated, and the root canals were instrumented to ISO size 50 at the working length. The apical 3 mm of the roots were resected, and cracks were artificially created in the apical dentin with an average load of 5.6 kg using a cylindrical wedge in a miniature drill press. A video microscope at x65 magnification was used to observe the cracks as they developed. Four independent examiners evaluated the root specimens using unaided/corrected vision (group 1), loupes at x3.3 magnification (group 2), a surgical operating microscope at x10 magnification (group 3), and the Orascope at x35 magnification (group 4). The examiners' proficiency at correctly identifying root ends with and without cracks was evaluated. The data were compared to the predetermined standard (27 cracked, 23 not cracked) with a one-tailed Fisher's exact test (alpha = 0.05). Statistically, the Orascope (p = 0.02) was significantly superior, whereas using unaided/corrected vision (p = 0.99), loupes (p = 0.88), or the microscope (p = 0.14) was not significantly better than guessing. The accuracy of correct identification for unaided/ corrected vision, loupes, the microscope, and the Orascope was 39%, 45%, 53%, and 58%, respectively. A two-way analysis of variance of the accuracy of crack identification showed a significant difference among the four visualization techniques (p = 0.0007) and also among the four evaluators (p = 0.006).


Assessment and management of behavioral and psychological symptoms of dementia.
Behavioral and psychological symptoms of dementia include agitation, depression, apathy, repetitive questioning, psychosis, aggression, sleep problems, wandering, and a variety of inappropriate behaviors. One or more of these symptoms will affect nearly all people with dementia over the course of their illness. These symptoms are among the most complex, stressful, and costly aspects of care, and they lead to a myriad of poor patient health outcomes, healthcare problems, and income loss for family care givers. The causes include neurobiologically related disease factors; unmet needs; care giver factors; environmental triggers; and interactions of individual, care giver, and environmental factors. The complexity of these symptoms means that there is no "one size fits all solution," and approaches tailored to the patient and the care giver are needed. Non-pharmacologic approaches should be used first line, although several exceptions are discussed. Non-pharmacologic approaches with the strongest evidence base involve family care giver interventions. Regarding pharmacologic treatments, antipsychotics have the strongest evidence base, although the risk to benefit ratio is a concern. An approach to integrating non-pharmacologic and pharmacologic treatments is described. Finally, the paradigm shift needed to fully institute tailored treatments for people and families dealing with these symptoms in the community is discussed.


Patient with Severe Skeletal Class II Malocclusion: Double Jaw Surgery with Multipiece Le Fort I.
A 22-year-old woman with severe skeletal Class II malocclusion was referred to our clinic. A clinical examination revealed a convex soft tissue profile and increased teeth and gingiva exposure both while smiling and in the natural rest position. She had Class II molar and canine relationship with increased overjet, moderate crowding in both upper and lower jaws, and proclined upper and lower incisors. Skeletally, she showed transverse maxillary deficiency, maxillary vertical excess, and mandibular retrognathia. We planned orthodontic-orthognathic surgery with multipiece Le Fort I osteotomy and bilateral sagittal split osteotomy (BSSO) to achieve ideal occlusion, stability, and facial esthetics. During orthodontic decompensation to relieve the crowding and to gain an ideal incisor inclination, four bicuspid extractions were performed. Because we used continuous mechanics, at the end of the decompensation period, we cut the maxillary arch wire distal to the lateral incisors into three pieces and waited for 3 months for vertical and transversal dental relapse. During the double jaw surgical procedure, the maxilla expanded and impacted with multisegmented Le Fort I osteotomy and the mandible advanced with BSSO. After the orthodontic and orthognathic surgical treatment, the skeletal and dental imbalance was corrected, and functional occlusion and dental and skeletal Class I relationship were achieved. The treatment results were stable at the 1-year follow-up.


Minus-end capture of preformed kinetochore fibers contributes to spindle morphogenesis.
Near-simultaneous three-dimensional fluorescence/differential interference contrast microscopy was used to follow the behavior of microtubules and chromosomes in living alpha-tubulin/GFP-expressing cells after inhibition of the mitotic kinesin Eg5 with monastrol. Kinetochore fibers (K-fibers) were frequently observed forming in association with chromosomes both during monastrol treatment and after monastrol removal. Surprisingly, these K-fibers were oriented away from, and not directly connected to, centrosomes and incorporated into the spindle by the sliding of their distal ends toward centrosomes via a NuMA-dependent mechanism. Similar preformed K-fibers were also observed during spindle formation in untreated cells. In addition, upon monastrol removal, centrosomes established a transient chromosome-free bipolar array whose orientation specified the axis along which chromosomes segregated. We propose that the capture and incorporation of preformed K-fibers complements the microtubule plus-end capture mechanism and contributes to spindle formation in vertebrates.


Emergence of an early SARS-CoV-2 epidemic in the United States.
The emergence of the COVID-19 epidemic in the United States (U.S.) went largely undetected due to inadequate testing. New Orleans experienced one of the earliest and fastest accelerating outbreaks, coinciding with Mardi Gras. To gain insight into the emergence of SARS-CoV-2 in the U.S. and how large-scale events accelerate transmission, we sequenced SARS-CoV-2 genomes during the first wave of the COVID-19 epidemic in Louisiana. We show that SARS-CoV-2 in Louisiana had limited diversity compared to other U.S. states and that one introduction of SARS-CoV-2 led to almost all of the early transmission in Louisiana. By analyzing mobility and genomic data, we show that SARS-CoV-2 was already present in New Orleans before Mardi Gras, and the festival dramatically accelerated transmission. Our study provides an understanding of how superspreading during large-scale events played a key role during the early outbreak in the U.S. and can greatly accelerate epidemics.


The thrombogenic effect of anticancer drug therapy in women with stage II breast cancer.
Thromboembolic disease has long been recognized as a complication of cancer. Recent reports have suggested that drugs used in the treatment of cancer, including chemotherapeutic agents and hormones, may contribute to this risk, but it has not been possible to separate the effect of these drugs from that of the cancer. We performed a randomized trial comparing 12 weeks of chemohormonal therapy (using cyclophosphamide, methotrexate, fluorouracil, vincristine, prednisone, doxorubicin, and tamoxifen) with 36 weeks of chemotherapy (using cyclophosphamide, methotrexate, fluorouracil, vincristine, and prednisone) in patients with Stage II breast cancer. Among 205 patients randomly assigned to treatment, there were 14 episodes of thrombosis (6.8 percent). These 14 episodes occurred during 979 patient-months of chemotherapy; by comparison, there were no events during 2413 patient-months without therapy. During the first 12 weeks of the study, five patients in the 12-week group and four patients in the 36-week group had thrombosis. During the subsequent 24 weeks, when only patients in the 36-week group were still receiving chemotherapy, there was no thrombosis in the 12-week group, but there were five additional events in the 36-week group (P = 0.03). These findings suggest that chemotherapy contributes to thrombosis in patients with breast cancer.


The impact of anti-vascular endothelial growth factor agents on visual impairment/blindness prevention in patients with diabetic macular edema and on associated patient and caregiver burden in Japan.
To estimate the impact of anti-vascular endothelial growth factor (VEGF) agents on visual impairment and blindness avoided in patients with diabetic macular edema (DME) and on associated patient and caregiver productivity loss in Japan. This study compared the impact of current care (estimated at 53.8% utilization of anti-VEGF agents using current data) with that of hypothetical care (characterized by a higher utilization of anti-VEGF agents [80.0%], as estimated by an expert panel) of DME patients. A population-based Markov model (two-eye approach) simulated visual acuity (Early Treatment Diabetic Retinopathy Study [ETDRS] letters) transitions over 5 years with DME treatments (intravitreal aflibercept, ranibizumab, and triamcinolone acetonide, and grid/focal laser) in patients with DME. Patient and caregiver productivity loss was determined using the human capital method. In total, 570,000 DME patients were included in the model over 5 years. Increased utilization of anti-VEGF agents resulted in 6,659 fewer cases of severe visual impairment (SVI; 26-35 ETDRS letters) or blindness (0-25 ETDRS letters) compared with the current care approach (26,023 vs 32,682 cases; 20.38% reduction) over this period. Increased utilization of anti-VEGF agents also contributed to productivity loss savings of ¥12.58 billion (US $115.64 million) (i.e., 17.01%) at the end of year 5. The total overall saving over 5 years was ¥45.83 billion (US $421.27 million) (13.45%). Few Japanese data were available, and assumptions were made for some inputs. Vision changes dependent on the function of both eyes were not studied. Only intravitreal (not sub-Tenon's) injections of triamcinolone were considered in this model. Direct costs were not considered. Increased utilization of anti-VEGF agents can reduce SVI and legal blindness in patients with DME in Japan. This would also be associated with substantial savings in patient and caregiver productivity loss.


Low body mass index is associated with an increased risk of neuropsychiatric adverse events and concentration impairment in women on mefloquine.
We performed a prospective cohort study to gain more insight into risk factors for neuropsychiatric effects of mefloquine among tourists travelling to tropical areas. We enrolled all patients who consulted the Travel Clinic of the Havenziekenhuis & Institute for Tropical Diseases Rotterdam for mefloquine prophylaxis during the period between 1 May 1999 and 7 March 2000. Each patient was followed from baseline (prior to starting mefloquine) up to 3 weeks after starting weekly intake of 250 mg mefloquine. We compared the intraindividual change in scores between baseline and follow-up visit on the Dutch shortened Profile of Mood States, and on the Continuous Performance Test (CPT) which measures sustained attention. The final cohort consisted of 151 subjects with a mean age of 38 years. In this population, a significant impairment of mood state was observed in those with a body mass index (BMI) < or = 20 kg m(-2). Stratification for gender showed that the total mood disturbance in females in the lowest BMI category significantly increased by 8.42 points [95% confidence interval (CI) 3.33, 13.50], whereas BMI did not affect the risk in males. Stratification for history of use of mefloquine showed that the risks were highest in first-time users. Analyses of the CPT showed that reaction time in women with a BMI < or = 20 kg m(-2) increased significantly by 22.5 ms (95% CI 7.80, 37.20), whereas reaction time in men showed a slight and nonsignificant decrease. Risk factors for mefloquine-associated neuropsychiatric adverse events and concentration impairment are female gender, low BMI, and first-time use. The frequency of neuropsychiatric effects is highest in women with a BMI < or = 20 kg m(-2).


A simple and efficient protocol for the mass propagation of Cymbidium mastersii: an ornamental orchid of Northeast India.
Cymbidium mastersii is an epiphytic orchid distributed mainly in Northeast India. Owing to its high commercial value in the floricultural industry, natural populations are under threat from over-exploitation. Mass propagation provides an alternative means of satisfying the demand. Unfortunately, conventional propagation is slow and difficult, suggesting in vitro methods for mass multiplication may be more appropriate. The objective of this study was to develop an efficient protocol. FOUR NUTRIENT MEDIA WERE EVALUATED FOR SEED GERMINATION AND EARLY PROTOCORM DEVELOPMENT: Murashige and Skoog (MS), half-strength MS, Knudson 'C' (KC), and Vacin and Went (VW). In addition, the effects of plant growth regulators 6-benzylaminopurine (BAP), kinetin (KN), α-naphthalene acetic acid (NAA) and indole-3-butyric acid (IBA) were studied alone and in combination. The maximum percentage seed germination (93.58 ± 0.56) was obtained in MS basal medium after 8-9 weeks of culture. Secondary protocorms (protocorm-like bodies) were developed from primary protocorms on MS medium fortified with different concentrations and combinations of cytokinins (BAP and KN) and auxins (NAA and IBA). The highest numbers of secondary protocorms (20.55 ± 0.62)/primary protocorms were obtained in MS medium supplemented with 5.0 µM BAP and 2.5 µM NAA. The most effective auxin source promoting root production (7.46 ± 0.09 per shoot) was 10.0 µM IBA. The plants were acclimatized effectively (survival percentage 88 %) in a greenhouse using a rooting medium of crushed sterile brick and charcoal (1: 1v/v) and vermicompost (leaf litter + cow dung, 1: 1v/v). An efficient protocol was established for in vitro propagation of C. mastersii using seed as the starting material. The percentage seed germination varied with the composition of the nutrient media and was highest in full-strength MS basal medium. The number of secondary protocorms that developed from primary protocorms was increased by the addition of 5.0 µM BAP and 2.5 µM NAA. In vitro raised plantlets acclimatized in a greenhouse were closely similar to the mother plants in morphology.


Acute myeloid leukemia immunopeptidome reveals HLA presentation of mutated nucleophosmin.
Somatic mutations in cancer are a potential source of cancer specific neoantigens. Acute myeloid leukemia (AML) has common recurrent mutations shared between patients in addition to private mutations specific to individuals. We hypothesized that neoantigens derived from recurrent shared mutations would be attractive targets for future immunotherapeutic approaches. Here we sought to study the HLA Class I and II immunopeptidome of thirteen primary AML tumor samples and two AML cell lines (OCI-AML3 and MV4-11) using mass spectrometry to evaluate for endogenous mutation-bearing HLA ligands from common shared AML mutations. We identified two endogenous, mutation-bearing HLA Class I ligands from nucleophosmin (NPM1). The ligands, AVEEVSLRK from two patient samples and C(cys)LAVEEVSL from OCI-AML3, are predicted to bind the common HLA haplotypes, HLA-A*03:01 and HLA-A*02:01 respectively. Since NPM1 is mutated in approximately one-third of patients with AML, the finding of endogenous HLA ligands from mutated NPM1 supports future studies evaluating immunotherapeutic approaches against this shared target, for this subset of patients with AML.


Neuroimaging for drug addiction and related behaviors.
In this review, we highlight the role of neuroimaging techniques in studying the emotional and cognitive-behavioral components of the addiction syndrome by focusing on the neural substrates subserving them. The phenomenology of drug addiction can be characterized by a recurrent pattern of subjective experiences that includes drug intoxication, craving, bingeing, and withdrawal with the cycle culminating in a persistent preoccupation with obtaining, consuming, and recovering from the drug. In the past two decades, imaging studies of drug addiction have demonstrated deficits in brain circuits related to reward and impulsivity. The current review focuses on studies employing positron emission tomography (PET), functional magnetic resonance imaging (fMRI), and electroencephalography (EEG) to investigate these behaviors in drug-addicted human populations. We begin with a brief account of drug addiction followed by a technical account of each of these imaging modalities. We then discuss how these techniques have uniquely contributed to a deeper understanding of addictive behaviors.


Relationship between colorectal cancer screening adherence and knowledge among vulnerable rural residents of Appalachian Kentucky.
Colorectal cancer (CRC) is 1 of the leading causes of cancer-related deaths among residents of rural Appalachia. Rates of guideline-consistent CRC screening in Appalachian Kentucky are suboptimal. This study sought to determine the relationship between CRC screening knowledge, specifically regarding recommended screening intervals, and receipt of screening among residents of rural Appalachian Kentucky. Residents of Appalachian Kentucky (n = 1096) between the ages of 50 and 76 years completed a telephone survey including questions on demographics, health history, and knowledge about CRC screening between November 20, 2009, and April 22, 2010. Although 67% of respondents indicated receiving screenings according to guidelines, respondents also demonstrated significant knowledge deficiencies about screening recommendations. Nearly half of respondents were unable to identify the recommended screening frequency for any of the CRC screening modalities. Accuracy about the recommended frequency of screening was positively associated with screening adherence. Enhanced educational approaches have the potential to increase CRC screening adherence in this population and reduce cancer mortality in this underserved region. Nurses play a critical role in patient education, which ultimately may increase screening rates. To fulfill this role, nurses should incorporate current recommendation about CRC screening into educational sessions. Advanced practices nurses in rural settings should also be aware of the increased vulnerability of their patient population and develop strategies to enhance awareness about CRC and the accompanying screening tests.


Proof of Concept of an Assistive Robotic Arm Control Using Artificial Stereovision and Eye-Tracking.
Assistive robotic arms have become popular to help users with upper limb disabilities achieve autonomy in their daily tasks, such as drinking and grasping objects in general. Usually, these robotic arms are controlled with an adapted joystick. Joysticks are user-friendly when it comes to a general approach to an object. However, they are not as intuitive when having to accurately approach an object, especially when obstacles are present. Alternatively, the combined use of artificial stereovision and eye-tracking seems to be a promising solution, as the user's vision is usually dissociated from their upper limb disability. Hence, the objective of this study was to develop a proof of concept for the control of an assistive robotic arm using a low-cost combination of stereovision and eye-tracking. Using the developed control system, a typically developed person was able to control the robotic arm successfully reaching and grasping an object for 92% of the trials without obstacles with an average time of 13.8 seconds. Then, another set of trials with one obstacle had a success rate of 91% with an average time of 17.3 seconds. Finally, the last set of trials with two obstacles had a success rate of 98% with an average time of 18.4 seconds. Furthermore, the cost of an eye-tracker and stereovision remains below 400$.


Rounding up the usual suspects: a standard target-gene approach for resolving the interfamilial phylogenetic relationships of ecribellate orb-weaving spiders with a new family-rank classification (Araneae, Araneoidea).
We test the limits of the spider superfamily Araneoidea and reconstruct its interfamilial relationships using standard molecular markers. The taxon sample (363 terminals) comprises for the first time representatives of all araneoid families, including the first molecular data of the family Synaphridae. We use the resulting phylogenetic framework to study web evolution in araneoids. Araneoidea is monophyletic and sister to Nicodamoidea rank. n. Orbiculariae are not monophyletic and also include the RTA clade, Oecobiidae and Hersiliidae. Deinopoidea is paraphyletic with respect to a lineage that includes the RTA clade, Hersiliidae and Oecobiidae. The cribellate orb-weaving family Uloboridae is monophyletic and is sister group to a lineage that includes the RTA Clade, Hersiliidae and Oecobiidae. The monophyly of most Araneoidea families is well supported, with a few exceptions. Anapidae includes holarchaeids but the family remains diphyletic even if Holarchaea is considered an anapid. The orb-web is ancient, having evolved by the early Jurassic; a single origin of the orb with multiple "losses" is implied by our analyses. By the late Jurassic, the orb-web had already been transformed into different architectures, but the ancestors of the RTA clade probably built orb-webs. We also find further support for a single origin of the cribellum and multiple independent losses. The following taxonomic and nomenclatural changes are proposed: the cribellate and ecribellate nicodamids are grouped in the superfamily Nicodamoidea rank n. (Megadictynidae rank res. and Nicodamidae stat. n.). Araneoidea includes 17 families with the following changes: Araneidae is re-circumscribed to include nephilines, Nephilinae rank res., Arkyidae rank n., Physoglenidae rank n., Synotaxidae is limited to the genus Synotaxus, Pararchaeidae is a junior synonym of Malkaridae (syn. n.), Holarchaeidae of Anapidae (syn. n.) and Sinopimoidae of Linyphiidae (syn. n.).


Relation of patient characteristics to cardiac ischemia during daily life activity (an Asymptomatic Cardiac Ischemia Pilot Data Bank Study).
Cardiac ischemia during daily life activities, detected by ambulatory electrocardiographic (AECG) monitoring has been associated with increased risk for adverse outcomes. Because daily life ischemia is usually asymptomatic, prevalence and descriptive data of patients with asymptomatic cardiac ischemia (ACI) are not well defined. Accordingly, patients screened for the Asymptomatic Cardiac Ischemia Pilot (ACIP) trial by 48-hour AECG monitoring were investigated to identify factors associated with ACI. A total of 1,820 patients with ischemia on a screening stress test and/or known or suspected coronary artery disease underwent AECG monitoring. Their mean age was 61 years, range 33 to 89; 83% were men and 81% were white. On AECG monitoring, ACI occurred in 897 patients (49%). There was a modest trend (p = 0.04) between increasing age and ACI prevalence. Increased risk for ACI was observed in patients reporting angina 6 weeks before screening (odds ratio 1.38, 99% confidence interval 1.08 to 1.77, p = 0.0008). There was a positive association between increases in heart rate during daily life and ACI prevalence (p < 0.0001). No daily, monthly, or seasonal variation in ACI prevalence was found, although ACI was more prevalent in northern than southern sites. In this group of clinically stable patients, selected on the basis of high risk for coronary artery disease, the prevalence of ACI was higher than expected from previous reports. Several readily available clinical characteristics (i.e., advanced age, recent angina, increased heart rate change with daily activity) were associated with significantly increased probability of ACI.


Elevated concentrations of defensins in bronchoalveolar lavage fluid in diffuse panbronchiolitis.
Human neutrophils contain three isoforms of antimicrobial and cytotoxic peptides in the azurophil granules, which belong to a family of mammalian neutrophil peptides named defensins. Here we investigate the role of these peptides in diffuse panbronchiolitis (DPB). Defensins (human neutrophil peptide-1, -2 and -3) were measured by radioimmunoassay in bronchoalveolar lavage fluid (BALF) of 30 patients with DPB, 16 patients with idiopathic pulmonary fibrosis (IPF) and 15 healthy adults. The concentration of defensins was higher in BALF of patients with DPB than in patients with IPF and healthy subjects. DPB and IPF patients also had significantly higher plasma concentrations of defensins than controls. In patients with DPB, BALF concentration of defensins correlated significantly with neutrophil count or BALF concentration of interleukin (IL)-8. Immunohistochemistry of open-lung biopsy specimens from four DPB patients showed localization of defensins in neutrophils and mucinous exudate in the airways, and on the surface of bronchiolar epithelial cells. In vitro studies showed an enhanced extracellular release of defensins following stimulation of neutrophils with phorbol myristate acetate, N-formyl-methionyl-leucyl-phenyalamine, and human recombinant IL-8. Treatment of DPB with macrolides for 6 months significantly reduced neutrophil count and concentrations of defensins and IL-8 in BALF. Our results indicate accumulation of neutrophil-derived defensins in the airway in diffuse panbronchiolitis, and suggest that defensins may be a marker of neutrophil activity in this disease.


Severe hyperglycorrhachia and status epilepticus after endoscopic aqueductoplasty: illustrative case.
While hypoglycorrhachia is observed and managed frequently, there are few reports in the literature of clinically significant hyperglycorrhachia after neurosurgery. Understanding the effects and management of severe hyperglycorrhachia is important to the neurosurgeon and neurocritical care teams who care for patients in these rare scenarios. The authors present the case of a 3-month-old male with congenital hydrocephalus who faced profound hyperglycorrhachia and status epilepticus after an endoscopic aqueductoplasty using an irrigant composed of lactated Ringer's solution with dextrose 5% in water. A multidisciplinary approach was developed to monitor and treat the patient's seizures and cerebrospinal fluid (CSF) osmolytes. This case provides several learning opportunities for understanding CSF physiology, pathogenesis of common brain injuries related to osmotic shifts and inflammatory states, as well as clinical management of hyperglycorrhachia. It also reiterates the significance of meticulous intraoperative assessment to avoid preventable medical errors.


Repurposing Drugs in Oncology (ReDO)-itraconazole as an anti-cancer agent.
Itraconazole, a common triazole anti-fungal drug in widespread clinical use, has evidence of clinical activity that is of interest in oncology. There is evidence that at the clinically relevant doses, itraconazole has potent anti-angiogenic activity, and that it can inhibit the Hedgehog signalling pathway and may also induce autophagic growth arrest. The evidence for these anticancer effects, in vitro, in vivo, and clinical are summarised, and the putative mechanisms of their action outlined. Clinical trials have shown that patients with prostate, lung, and basal cell carcinoma have benefited from treatment with itraconazole, and there are additional reports of activity in leukaemia, ovarian, breast, and pancreatic cancers. Given the evidence presented, a case is made that itraconazole warrants further clinical investigation as an anti- cancer agent. Additionally, based on the properties summarised previously, it is proposed that itraconazole may synergise with a range of other drugs to enhance the anti-cancer effect, and some of these possible combinations are presented in the supplementary materials accompanying this paper.


A prospective, longitudinal study of the impact of GJB2/GJB6 genetic testing on the beliefs and attitudes of parents of deaf and hard-of-hearing infants.
There are limited data on the impact of incorporating genetic counseling and testing into the newborn hearing screening process. We report on results from a prospective, longitudinal study to determine the impact of genetic counseling and GJB2/GJB6 genetic testing on parental knowledge, attitudes, and beliefs about genetic testing. One hundred thirty culturally hearing parents of 93 deaf or hard-of-hearing children ages 0-3 years primarily identified through newborn hearing screening received pre- and post-test genetic counseling for GJB2 and GJB6. Parents completed questionnaires following pre-test counseling, and 1- and 6-month post-test result disclosure. Results indicate that following pre-test counseling all parents perceived benefits to genetic testing. While parents who received positive results continued to perceive benefits from testing, perceived benefit declined among parents who received inconclusive or negative results. Parents did not perceive genetic testing as harmful following pre-test counseling or receipt of test results. Parents who received positive test results performed better in understanding recurrence and causation of their child's deafness and indicated greater interest in prenatal genetic testing than those who received inconclusive or negative test results. Parents felt that pediatricians and audiologists should inform parents of genetic testing availability; however, there was no consensus on timing of this discussion. Thus culturally hearing parents do not perceive genetic testing of their deaf or hard-of-hearing infants/toddlers as harmful; they feel that primary care providers should discuss genetic testing with them; and positive genetic test results with genetic counseling give rise to better understanding and perceived benefit than negative or inconclusive results.


Observation of plasmarons in quasi-freestanding doped graphene.
A hallmark of graphene is its unusual conical band structure that leads to a zero-energy band gap at a single Dirac crossing point. By measuring the spectral function of charge carriers in quasi-freestanding graphene with angle-resolved photoemission spectroscopy, we showed that at finite doping, this well-known linear Dirac spectrum does not provide a full description of the charge-carrying excitations. We observed composite "plasmaron" particles, which are bound states of charge carriers with plasmons, the density oscillations of the graphene electron gas. The Dirac crossing point is resolved into three crossings: the first between pure charge bands, the second between pure plasmaron bands, and the third a ring-shaped crossing between charge and plasmaron bands.


Age-related delay in information accrual for faces: evidence from a parametric, single-trial EEG approach.
In this study, we quantified age-related changes in the time-course of face processing by means of an innovative single-trial ERP approach. Unlike analyses used in previous studies, our approach does not rely on peak measurements and can provide a more sensitive measure of processing delays. Young and old adults (mean ages 22 and 70 years) performed a non-speeded discrimination task between two faces. The phase spectrum of these faces was manipulated parametrically to create pictures that ranged between pure noise (0% phase information) and the undistorted signal (100% phase information), with five intermediate steps. Behavioural 75% correct thresholds were on average lower, and maximum accuracy was higher, in younger than older observers. ERPs from each subject were entered into a single-trial general linear regression model to identify variations in neural activity statistically associated with changes in image structure. The earliest age-related ERP differences occurred in the time window of the N170. Older observers had a significantly stronger N170 in response to noise, but this age difference decreased with increasing phase information. Overall, manipulating image phase information had a greater effect on ERPs from younger observers, which was quantified using a hierarchical modelling approach. Importantly, visual activity was modulated by the same stimulus parameters in younger and older subjects. The fit of the model, indexed by R2, was computed at multiple post-stimulus time points. The time-course of the R2 function showed a significantly slower processing in older observers starting around 120 ms after stimulus onset. This age-related delay increased over time to reach a maximum around 190 ms, at which latency younger observers had around 50 ms time lead over older observers. Using a component-free ERP analysis that provides a precise timing of the visual system sensitivity to image structure, the current study demonstrates that older observers accumulate face information more slowly than younger subjects. Additionally, the N170 appears to be less face-sensitive in older observers.


[Genotyping of nosocomial methicillin-resistant Staphylococcus aureus strains isolated from clinical specimens by rep-PCR].
Methicillin-resistant Staphylococcus aureus (MRSA) infections are associated with increased cost, mortality and length of hospital stay compared with the other infections. Therefore, controlling the spread of this pathogen by screening patients, personnel and the environment remains as a high priority in infection control programs. The aim of this study was to detect the clonal relationship between nosocomial MRSA strains by using repetitive-sequence-based polymerase chain reaction (rep-PCR) method which has several advantages owing to its speed and ease of use. A total of 100 MRSA stock strains that had been isolated from various clinical samples of hospitalized patients in Erciyes University Medical Faculty Hospitals between September 2008-October 2009, were included in the study. Methicillin resistance of the strains were determined by cefoxitin disc diffusion test according to CLSI guidelines. Rep-PCR (Diversilab, bioMerieux, France) method was performed in the following four steps in order to determine genetic proximity of MRSA strains: (1) Manual DNA extraction (UltraClean Microbial DNA Isolation Kit; MoBio Laboratories, USA), (2) Rep-PCR by using fingerprinting kits in the thermocycler (Diversilab DNA Fingerprinting Kit), (3) Automated microfluidic electrophoresis by bioanalyzer (Diversilab DNA LabChip kit), (4) Analysis and rapid evaluation with the use of web-based DiversiLab software (version 2.1.66). Rep-PCR analysis have shown the presence of a total 11 clones, including 3 major clones [A (4 subtypes), B (2 subtypes) and C (2 subtypes)] and 8 unique clones (DK). Clone A was found to be the dominant type. Seventy-eight percent of the 100 MRSA isolates belonged to clone A (63 were A1; 9 were A2; 4 were A3, 2 were A4), 11% belonged to clone B (10 were B1, 1 was B2), 3% belonged to clone C (2 were C1, 1 was C2), and one of each belonged to the other clones (D, E, F, G, H, I, J, K). Clone A was isolated from 93.3% (14/15) of the samples sent from internal diseases intensive care unit (ICU), from 66.6% (10/15) of the samples sent from infectious diseases ward and 91% (10/11) of hematology-oncology ward samples. All MRSA strains isolated from anesthesiology and newborn ICU were of clone A. The isolation dates of these strains were in proximity. In conclusion, MRSA strains showed clonal dissemination in our hospital, clone A being the predominant one during the study period. Rep-PCR which is a rapid and reliable method, can easily be applied for molecular epidemiological purposes and aid to infection control measures.


The association of copy number variation and percent mammographic density.
Percent mammographic density (PD) estimates the proportion of stromal, fat, and epithelial breast tissues on the mammogram image. Adjusted for age and body mass index (BMI), PD is one of the strongest risk factors for breast cancer. Inherited factors are hypothesized to explain between 30 and 60% of the variance in this trait. However, previously identified common genetic variants account for less than 6% of the variance in PD, leaving much of the genetic contribution to this trait unexplained. We performed the first study to examine whether germline copy number variation (CNV) are associated with PD. Two genome-wide association studies (GWAS) of percent density conducted on the Illumina 660W-Quad were used to identify and replicate the association between candidate CNVs and PD: the Minnesota Breast Cancer Family Study (MBCFS) and controls from the Mayo Venous Thromboembolism (Mayo VTE) Case-Control Study, with 585 and 328 women, respectively. Linear models were utilized to examine the association of each probe with PD, adjusted for age, menopausal status and BMI. Segmentation was subsequently performed on the probe-level test statistics to identify candidate CNV regions that were associated with PD. Sixty-one probes from five chromosomal regions [3q26.1 (2 regions), 8q24.22, 11p15.3, and 17q22] were significantly associated with PD in MBCFS (p-values <0.0001). A CNV at 3q26.1 showed the greatest evidence for association with PD; a region without any known SNPs. Conversely, the CNV at 17q22 was largely due to the association between SNPs and PD in the region. SNPs in the 8q24.22 region have been shown to be associated with risk of many cancers; however, SNPs in this region were not responsible for the observed CNV association. While we were unable to replicate the associations with PD, two of the five CNVs (3q26.1 and 11p15.3) were also observed in the Mayo VTE controls. CNVs may help to explain some of the variability in PD that is currently unexplained by SNPs. While we were able to replicate the existence of two CNVs across the two GWAS studies, we were unable to replicate the associations with PD. Even so, the proximity of the identified CNV regions to loci known to be associated with breast cancer risk suggests further investigation and potentially shared genetic mechanisms underlying the PD and breast cancer association.


To tame a volcano: patients with borderline personality disorder and their perceptions of suffering.
The aim of the study was to investigate life situations, suffering, and perceptions of encounter with psychiatric care among 10 patients with borderline personality disorder. The results are based on a hermeneutic interpretation of narrative interviews in addition to biographical material (diary excerpts and poems). The interpretation revealed three comprehensive theme areas: life on the edge , the struggle for health and dignity-a balance act on a slack wire over a volcano , and the good and the bad act of psychiatric care in the drama of suffering . These theme areas form a movement back and forth-from despair and unendurable suffering to struggle for health and dignity and a life worth living. Common beliefs regarding these patients among personnel and implications for psychiatric care are discussed in relation to the results.


Unconscious Familiarity-based Color-Form Binding: Evidence from Visual Extinction.
There is good evidence that early visual processing involves the coding of different features in independent brain regions. A major question, then, is how we see the world in an integrated manner, in which the different features are "bound" together. A standard account of this has been that feature binding depends on attention to the stimulus, which enables only the relevant features to be linked together [Treisman, A., & Gelade, G. A feature-integration theory of attention. Cognitive Psychology, 12, 97-136, 1980]. Here we test this influential idea by examining whether, in patients showing visual extinction, the processing of otherwise unconscious (extinguished) stimuli is modulated by presenting objects in their correct (familiar) color. Correctly colored objects showed reduced extinction when they had a learned color, and this color matched across the ipsi- and contralesional items (red strawberry + red tomato). In contrast, there was no reduction in extinction under the same conditions when the stimuli were colored incorrectly (blue strawberry + blue tomato; Experiment 1). The result was not due to the speeded identification of a correctly colored ipsilesional item, as there was no benefit from having correctly colored objects in different colors (red strawberry + yellow lemon; Experiment 2). There was also no benefit to extinction from presenting the correct colors in the background of each item (Experiment 3). The data suggest that learned color-form binding can reduce extinction even when color is irrelevant for the task. The result is consistent with preattentive binding of color and shape for familiar stimuli.


CMT3 alters mitochondrial function in murine osteoclast lineage cells.
Chemically modified tetracyclines (CMTs 1-10) were developed as non-antibiotic inhibitors of matrix metalloproteinases (MMPs). We previously demonstrated that MMP inhibition alone is insufficient to explain the pro-apoptotic action of CMTs in osteoclast lineage cells and we have explored additional mechanisms of action. We compared the characteristics of apoptosis in RAW264.7 murine monocyte and osteoclast cultures treated with pharmacologically relevant concentrations of CMT3 or the bisphosphonate alendronate, which induces osteoclast apoptosis through inhibition of farnesyl diphosphate synthase. CMT3 induced apoptosis rapidly (2-3h), whereas alendronate-induced apoptosis was delayed (>12h). CMT3-treated cells did not accumulate unprenylated Rap1A in contrast to cells treated with alendronate. Importantly, CMT3 induced a rapid loss of mitochondrial stability in RAW264.7 cells measured by loss of Mitotracker((R)) Red fluorescence, while bongkrekic acid protected polykaryons from CMT3-induced apoptosis. Modulation of mitochondrial function is therefore a significant early action of CMT3 that promotes apoptosis in osteoclast lineage cells.


[Perineal Groove: A Rare Congenital Anomaly].
Perineal groove is a rare congenital anorectal malformation, with incidence yet undetermined. It is almost exclusive to the female newborn and its embryogenic origin remains uncertain. We present a case-report of a newborn girl that was discharged from the nursery without complications. At her first appointment at primary care we noted a wet sulcus connecting the posterior vaginal commissure and the anus. This case report emphasizes the rarity of the perineal groove and the importance of a good quality history and physical examination at primary care.


A qualitative study of the dissemination and diffusion of innovations: bottom up experiences of senior managers in three health districts in South Africa.
In 2012 the South African National Department of Health (SA NDoH) set out, using a top down process, to implement several innovations in eleven health districts in order to test reforms to strengthen the district health system. The process of disseminating innovations began in 2012 and senior health managers in districts were expected to drive implementation. The research explored, from a bottom up perspective, how efforts by the National government to disseminate and diffuse innovations were experienced by district level senior managers and why some dissemination efforts were more enabling than others. A multiple case study design comprising three cases was conducted. Data collection in 2012 - early 2014 included 38 interviews with provincial and district level managers as well as non- participant observation of meetings. The Greenhalgh et al. (Milbank Q 82(4):581-629, 2004) diffusion of innovations model was used to interpret dissemination and diffusion in the districts. Managers valued the national Minister of Health's role as a champion in disseminating innovations via a road show and his personal participation in an induction programme for new hospital managers. The identification of a site coordinator in each pilot site was valued as this coordinator served as a central point of connection between networks up the hierarchy and horizontally in the district. Managers leveraged their own existing social networks in the districts and created synergies between new ideas and existing working practices to enable adoption by their staff. Managers also wanted to be part of processes that decide what should be strengthened in their districts and want clarity on: (1) the benefits of new innovations (2) total funding they will receive (3) their specific role in implementation and (4) the range of stakeholders involved. Those driving reform processes from 'the top' must remember to develop well planned dissemination strategies that give lower-level managers relevant information and, as part of those strategies, provide ongoing opportunities for bottom up input into key decisions and processes. Managers in districts must be recognised as leaders of change, not only as implementers who are at the receiving end of dissemination strategies from those at the top. They are integral intermediaries between those at the at the coal face and national policies, managing long chains of dissemination and natural (often unpredictable) diffusion.


Fluctuations in the emotional intelligence of therapy students during clinical placements: Implication for educators, supervisors, and students.
This study investigated the changes in emotional intelligence (EI) of occupational therapy, physiotherapy, and speech pathology students (therapy students). Clinical placements have multiple benefits including the development of interprofessional skills, enhancing practice skills and interpersonal skills. Higher EI competencies have been shown to have a positive impact on patient outcomes, teamwork skills, dealing with stress, and patient satisfaction. Data for this study were collected at two time points: before third-year therapy students commenced extended clinical placements (T1 with 261 students) and approximately 7 months later after students had completed one or more clinical placements (T2 with 109 students). EI was measured using the Emotional Quotient Inventory 2.0 (EQ-i2.0). Only one EI score, assertiveness, demonstrated a significant decline. No EI score showed a significant increase. A third or more of the students showed increases of five points or more in self-actualisation, emotional expression, independence, reality testing and optimism. However, of concern were the five EI scores where therapy students' EI scores decreased by more than five points: assertiveness (where 38% of students declined), problem solving (37%), impulse control (35%), self-actualisation (35%), and stress tolerance (33%). With EI scores declining for some students during clinical placements, there are implications for clinical supervisors and interprofessional facilitators as clinical performance may decline concurrently. There is a range of potential reasons that clinical placements could negatively influence the EI competencies of a therapy student, including poor clinical supervision, conflict between a student, and supervisor and failing a clinical placement. The research suggests that interprofessional facilitators and university educators might consider students undertaking EI tests before clinical placements.


Association between tobacco and alcohol use among young adult bar patrons: a cross-sectional study in three cities.
Bars and nightclubs are key public venues where young adults congregate and use both tobacco and alcohol, and young adult bar patrons are at high risk for substance use. This study examined the association between cigarette smoking and alcohol use among a random sample of young adult bar patrons from three different cities in the USA. Cross-sectional data was collected from a random sample of young adult bar patrons aged 18-29 in San Diego, CA (N = 1,150), Portland, ME (N = 1,019), and Tulsa, OK (N = 1,106) from 2007-2010 (response rate 88%) using randomized time location sampling. Respondents reported the number of days they smoked cigarettes, drank alcohol, and binge drank in the past 30 days. Multinomial logistic regression was used to analyze the association between smoking (nonsmoker, occasional smoker, and regular smoker) and drinking and binge drinking for each city controlling for age, gender, race/ethnicity, and education. Predicted probabilities of each smoking category were calculated by drinking and binge drinking status. The association between smoking and drinking and binge drinking among combined samples was also analyzed, controlling for demographic variables and city. Respondents reported high current smoking rates, ranging from 51% in Portland to 58% in Tulsa. Respondents in Tulsa were more likely to report regular smoking than those in San Diego and Portland, with demographic variables being controlled. Young adult bar patrons also exhibited a strong association between smoking and drinking. In general, as the frequency of drinking and binge drinking increased, the predicted probability of being a smoker, especially a regular smoker, increased in each city. Young adult bar patrons consistently reported a high smoking rate and a strong relationship between smoking and drinking, regardless of the different bar cultures and tobacco control contexts in each of the three cities. While smoke-free bar policies were negatively associated with regular smoking, these policies alone may not be enough to influence the association between smoking and drinking, particularly if tobacco marketing continues in these venues, or in the absence of programs specifically addressing the co-use of tobacco and alcohol.


Management of epistaxis.
Family physicians frequently encounter patients with epistaxis (nasal bleeding). In rare cases, this condition may lead to massive bleeding and even death. Although epistaxis can have an anterior or posterior source, it most often originates in the anterior nasal cavity. A directed history and physical examination generally determine the cause of the bleeding. Both local and systemic processes can play a role in epistaxis. Nasal bleeding usually responds to first-aid measures such as compression. When epistaxis does not respond to simple measures, the source of the bleeding should be located and treated appropriately. Treatments to be considered include topical vasoconstriction, chemical cautery, electrocautery, nasal packing (nasal tampon or gauze impregnated with petroleum jelly), posterior gauze packing, use of a balloon system (including a modified Foley catheter), and arterial ligation or embolization. Topical or systemic antibiotics should be used in selected patients. Hospital admission should be considered for patients with significant comorbid conditions or complications of blood loss. Referral to an otolaryngologist is appropriate when bleeding is refractory, complications are present, or specialized treatment (balloon placement, arterial ligation, angiographic arterial embolization) is required.


Surprising gene expression patterns within and between PDF-containing circadian neurons in Drosophila.
To compare circadian gene expression within highly discrete neuronal populations, we separately purified and characterized two adjacent but distinct groups of Drosophila adult circadian neurons: the 8 small and 10 large PDF-expressing ventral lateral neurons (s-LNvs and l-LNvs, respectively). The s-LNvs are the principal circadian pacemaker cells, whereas recent evidence indicates that the l-LNvs are involved in sleep and light-mediated arousal. Although half of the l-LNv-enriched mRNA population, including core clock mRNAs, is shared between the l-LNvs and s-LNvs, the other half is l-LNv- and s-LNv-specific. The distribution of four specific mRNAs is consistent with prior characterization of the four encoded proteins, and therefore indicates successful purification of the two neuronal types. Moreover, an octopamine receptor mRNA is selectively enriched in l-LNvs, and only these neurons respond to in vitro application of octopamine. Dissection and purification of l-LNvs from flies collected at different times indicate that these neurons contain cycling clock mRNAs with higher circadian amplitudes as well as at least a 10-fold higher fraction of oscillating mRNAs than all previous analyses of head RNA. Many of these cycling l-LNv mRNAs are well expressed but do not cycle or cycle much less well elsewhere in heads. The results suggest that RNA cycling is much more prominent in circadian neurons than elsewhere in heads and may be particularly important for the functioning of these neurons.


Protocol and quality assurance for carotid imaging in 100,000 participants of UK Biobank: development and assessment.
Background Ultrasound imaging is able to quantify carotid arterial wall structure for the assessment of cerebral and cardiovascular disease risks. We describe a protocol and quality assurance process to enable carotid imaging at large scale that has been developed for the UK Biobank Imaging Enhancement Study of 100,000 individuals. Design An imaging protocol was developed to allow measurement of carotid intima-media thickness from the far wall of both common carotid arteries. Six quality assurance criteria were defined and a web-based interface (Intelligent Ultrasound) was developed to facilitate rapid assessment of images against each criterion. Results and conclusions Excellent inter and intra-observer agreements were obtained for image quality evaluations on a test dataset from 100 individuals. The image quality criteria then were applied in the UK Biobank Imaging Enhancement Study. Data from 2560 participants were evaluated. Feedback of results to the imaging team led to improvement in quality assurance, with quality assurance failures falling from 16.2% in the first two-month period examined to 6.4% in the last. Eighty per cent had all carotid intima-media thickness images graded as of acceptable quality, with at least one image acceptable for 98% of participants. Carotid intima-media thickness measures showed expected associations with increasing age and gender. Carotid imaging can be performed consistently, with semi-automated quality assurance of all scans, in a limited timeframe within a large scale multimodality imaging assessment. Routine feedback of quality control metrics to operators can improve the quality of the data collection.


Measurement properties of the Minimal Insomnia Symptom Scale (MISS) in an elderly population in Sweden.
Insomnia is common among elderly people and associated with poor health. The Minimal Insomnia Symptom Scale (MISS) is a three item screening instrument that has been found to be psychometrically sound and capable of identifying insomnia in the general population (20-64 years). However, its measurement properties have not been studied in an elderly population. Our aim was to test the measurement properties of the MISS among people aged 65 + in Sweden, by replicating the original study in an elderly sample. Data from a cross-sectional survey of 548 elderly individuals were analysed in terms of assumptions of summation of items, floor/ceiling effects, reliability and optimal cut-off score by means of ROC-curve analysis and compared with self-reported insomnia criteria. Corrected item-total correlations ranged between 0.64-0.70, floor/ceiling effects were 6.6/0.6% and reliability was 0.81. ROC analysis identified the optimal cut-off score as ≥ 7 (sensitivity, 0.93; specificity, 0.84; positive/negative predictive values, 0.256/0.995). Using this cut-off score, the prevalence of insomnia in the study sample was 21.7% and most frequent among women and the oldest old. Data support the measurement properties of the MISS as a possible insomnia screening instrument for elderly persons. This study make evident that the MISS is useful for identifying elderly people with insomnia-like sleep problems. Further studies are needed to assess its usefulness in identifying clinically defined insomnia.


Graft-vs.-host-associated immune suppression is activated by recognition of allogeneic murine I-A antigens.
Several combinations of F1 hybrid mice were injected intravenously with parental spleen cells to determine the minimal H-2 differences between F1 and parent that are necessary to induce graft-vs.-host-associated immune suppression (GVH-associated suppression). 7-14 d after injection, the spleens of the F1 mice were tested for cytotoxic T lymphocyte potential by in vitro sensitization against trinitrophenyl-self and H-2 alloantigens. The results indicate that parental T lymphocytes must recognize I-A allogeneic determinants of the F1 recipient in order to induce suppression. Recognition of K or D alone or D with I region products other than I-A did not induce suppression. The recognition of I region without K and/or D and even the I-A difference between C57BL/6 and the B6.Cbm12 mutation resulted in immune suppression that was as potent as that resulting from the recognition of K, D, and I together. The possible significance of this function for I-A antigens is discussed with respect to three clinical examples of immune suppression for which this phenomenon may be relevant.


Incorporation of a laminin-derived peptide (SIKVAV) on polymer-modified adenovirus permits tumor-specific targeting via alpha6-integrins.
Effective gene therapy for disseminated metastatic cancer is currently impossible because of poor delivery of vector to target sites. Modification of viral vectors to target advanced cancer has long been a challenge. In this study, we aimed to redirect adenovirus tropism to infect prostate cancer cells via alpha6beta1 integrins, whose expression is upregulated during prostate cancer progression. To ablate normal mechanisms of infection and provide a framework for attachment of targeting ligands, viruses were non-genetically modified with pHPMA-ONp polymer. Addition of polymer-coated virus to prostate cells showed significantly reduced transgene expression compared with unmodified virus. To restore infectivity, an alpha6-integrin binding peptide (-SIKVAV-) derived from laminin was incorporated onto the surface of the polymer-coated viruses. Photon correlation spectroscopic analysis revealed a small increase in the mean diameter of the particles following retargeting. Addition of -SIKVAV- peptide restored virus infectivity of PC-3 cells in a ligand concentration-dependent manner that was significantly improved following removal of unincorporated polymer and peptide. Competition assays using cells preincubated with Ad5 fiber protein or free -SIKVAV- peptide confirmed that entry of retargeted viruses was mediated via the incorporated ligand. Application of retargeted viruses to a panel of human cell lines revealed varying levels of transduction efficiency. Flow cytometric analysis of cells using anti-alpha6 integrin and anti-beta1 integrin antibodies demonstrated that for prostate cells, greater transduction efficiency correlated with higher levels of expression of both integrin subunits. Furthermore with the exception of LNCaP cells, increased alpha6beta1 integrin expression correlated with advanced disease. Intravenous administration of retargeted viruses to tumor-bearing mice resulted in slower plasma clearance and greatly reduced liver tropism, and hence toxicity compared with unmodified virus, while maintaining reporter gene expression in the tumor. The data suggest that YESIKVAVS-retargeted viruses have potential for systemic delivery for the treatment of metastatic disease.


Percutaneous Mitral Valve Interventions (Repair): Current Indications and Future Perspectives.
Mitral valve regurgitation (MR) is the commonest valvular abnormality encountered among adult patients with cardiac valvular disease and conveys significant morbidity and mortality. The mitral valve is a complex anatomical structure and etiology for regurgitation is classified as either primary or secondary MR. Identification of the etiology in severe MR is critical in determining the appropriate treatment strategy. Transcatheter mitral valve repair (TMVR) is a minimally invasive technique for treatment of selected patients with symptomatic chronic moderate-severe or severe (3 to 4+) MR. While surgery remains the mainstay for treatment in primary MR, several technological advances within the last decade have made transcatheter mitral valve intervention increasingly feasible and safe in clinical practice. Use of TMVR in patients with severe MR has successfully reduced patient symptoms, disease morbidity, improved quality of life, and facilitated reverse remodeling with potential for a survival advantage among certain patients with secondary MR. Recent randomized controlled trials on MitraClip use in secondary MR have reinvigorated interest in this disease and refocused our attention on optimizing patient selection and timing of intervention to maximize benefit from using such percutaneous devices. In our review, we discuss etiologies and pathophysiology in both acute MR and development of chronic severe MR. We discuss management strategies for MR among patients based on etiology, particularly percutaneous mitral valve interventional therapies. We perform an extensive review comparing and contrasting existing data on safety, efficacy, durability, and appropriate patient selection related to MitraClip implantation in both primary and secondary MR. Lastly, we explore percutaneous MV therapies beyond the MitraClip as we await larger scale trials on these devices prior to them making way into day-to-day practice.


Spinal cord injury and traumatic brain injury: a cost-of-illness study.
Among traumatic injuries, spinal cord injuries (SCI) and traumatic brain injuries (TBI) are of major importance because of their epidemiological and economic impact on society. The overall objective of this study was to estimate the economic cost associated with people with SCI and TBI in Spain in 2007. A cost-of-illness analysis was performed, considering the perspective of society, using a 1-year time horizon. Medical costs, adaptation costs, material costs, administrative costs, and costs of police, firefighters and roadside assistance, productivity losses due to institutionalization and sick leave, as well as an estimate of productivity losses of carers, and productivity losses due to death were included. The economic cost associated with people with SCI is between EUR 92,087,080.97 and 212,496,196.41 (USD 131 million and 302 million) according to the injury mechanism, and between EUR 1,079,223,688.66 and 3,833,752,692.78 (USD 1,536 million and 5,458 million) for people with TBI. There is an urgent need to develop effective interventions known to prevent SCI and TBI, and to evaluate their effectiveness and efficiency.


A unique role for 6-O sulfation modification in zebrafish vascular development.
Heparan sulfate proteoglycans are important modulators of growth factor signaling in a variety of patterning processes. Secreted growth factors that play critical roles in angiogenesis bind to heparan sulfate, and this association is affected by 6-O-sulfation of the heparan sulfate chains. Addition of 6-O-sulfate is catalyzed by a family of sulfotransferases (HS6STs), and genetic manipulation of their function permits an assessment of their contribution to vascular assembly. We report on the biochemical activity and expression patterns of two zebrafish HS6ST genes. In situ hybridization reveals dynamic and distinct expression patterns of these two genes during development. Structural analysis of heparan sulfate from wild-type and morpholino antisense 'knockdown' embryos suggests that HS6ST-1 and HS6ST-2 have similar biochemical activity. HS6ST-2, but not HS6ST-1, morphants exhibit abnormalities in the branching morphogenesis of the caudal vein during embryonic development of the zebrafish. Our finding that HS6ST-2 is required for the branching morphogenesis of the caudal vein is the first in vivo evidence for an essential role of a gene encoding a heparan sulfate modifying enzyme in vertebrate angiogenesis. Our analysis of two zebrafish HS6ST genes suggests that a wide range of biological processes may be regulated by an array of sulfation-modifying enzymes in the vertebrate genome.


Effect of repeated invitations on uptake of colorectal cancer screening using faecal occult blood testing: analysis of prevalence and incidence screening.
To analyse the effects of prevalence and incidence screening on uptake and detection of cancer in an ongoing, dynamic programme for colorectal screening using faecal occult blood testing. Analysis of prevalence and incidence screening. Three rounds of biennial colorectal screening using the guaiac faecal occult blood test in east and north east Scotland, March 2000 to May 2007. Adults aged 50-69. Uptake of screening, test positivity (percentage of those invited who returned a test that was positive and triggered an invitation for colonoscopy), positive predictive value, and stage of cancer. Of 510 990 screening episodes in all three rounds, 248 998 (48.7%) were for prevalence, 163 483 (32.0%) were for first incidence, and 98 509 (19.3%) were for second incidence. Uptake of a first invitation for prevalence screening was 53% and for a second and third invitation was 15% and 12%. In the cohort invited for the first round, uptake of prevalence screening rose from 55% in the first round to 63% in the third. The uptake of first incidence screening on a first invitation was 54% and on a second invitation was 86% and on a first invitation for second incidence screening was 46%. The positivity rate in prevalence screening was 1.9% and the uptake of colonoscopy was 87%. The corresponding values for a first incidence screen were 1.7% and 90% and for a second incidence screen were 1.1% and 94.5%. The positive predictive value of a positive faecal occult blood test result for cancer was 11.0% for prevalence screening, 6.5% for the first incidence screen, and 7.5% for the second incidence screen. The corresponding values for the positive predictive value for adenoma were 35.5%, 29.4%, and 26.7%. The proportion of cancers at stage I dropped from 46.5% for prevalence screening to 41% for first incidence screening and 35% for second incidence screening. Repeat invitations to those who do not take up the offer of screening increases the number of those who accept, for both prevalence screening and incidence screening. Although the positive predictive value for both cancer and adenomas fell between the prevalence screen and the first incidence screen, they did not fall between the first and second incidence screens. The deterioration in cancer stage from prevalence to incidence screening suggests that some cancers picked up at incidence screening may have been missed on prevalence screening, but the stage distribution is still favourable. These data vindicate the policies of continuing to offer screening to those who fail to participate and continuing to offer biennial screening to those who have accepted previous offers.


Sympatric speciation suggested by monophyly of crater lake cichlids.
The existence of sympatric speciation--that populations diverge into species in the absence of physical or ecological barriers--is controversial. The East African Great Lakes harbour hundreds of cichlid species representing only a few monophyletic lineages, although palaeolimnological evidence and local restrictions on species distribution suggest that speciation in these lakes could have been allopatric. The case for sympatry in restricted areas of Lakes Malawi and Tanganyika is stronger but not unassailable. A better case might be made for cichlid species flocks in small, ecologically monotonous crater lakes. Here we present a mitochondrial DNA analysis of cichlid species flocks endemic to two such lakes in Cameroon. The results suggest that the flocks in each lake are monophyletic: the implication being that each lake was colonized once only, the size and shape of each lake being such that subsequent diversification would have been sympatric.


Photochemistry in a soft-glass single-ring hollow-core photonic crystal fibre.
A hollow-core photonic crystal fibre (HC-PCF), guided by photonic bandgap effects or anti-resonant reflection, offers strong light confinement and long photochemical interaction lengths in a microscale channel filled with a solvent of refractive index lower than that of glass (usually fused silica). These unique advantages have motivated its recent use as a highly efficient and versatile microreactor for liquid-phase photochemistry and catalysis. In this work, we use a single-ring HC-PCF made from a high-index soft glass, thus enabling photochemical experiments in higher index solvents. The optimized light-matter interaction in the fibre is used to strongly enhance the reaction rate in a proof-of-principle photolysis reaction in toluene.


Vancomycin-tolerant Streptococcus pneumoniae in Korea.
A nationwide surveillance study was undertaken to monitor antimicrobial resistance among clinical isolates of Streptococcus pneumoniae in Korea, with a special focus on vancomycin tolerance. For the 6-month period from March to August 2002, clinical isolates of S. pneumoniae were collected from 11 university hospitals and 1 reference laboratory. One-hundred eighty-eight isolates were measured for lysis rates after exposure to vancomycin for 4 h. Two vancomycin-tolerant S. pneumoniae (VTSP) strains, S3 and H8, were isolated from sputum cultures of two patients, who had stayed in intensive-care units of different hospitals with long-term antibiotic therapy and were not treated for pneumococcal pneumonia. The penicillin, cefotaxime, and vancomycin MICs for S3 were 8 microg/ml, >16 microg/ml, and 0.5 microg/ml, and those for H8 were 2 microg/ml, 2 microg/ml, and 0.5 microg/ml, respectively. While S3 belonged to serotype 23F and was autolysin defective, H8 belonged to serotype 13F and had intact autolysin. These strains were not clonally related as determined by pulsed-field gel electrophoresis of chromosomal DNA. In agreement with previous reports, both isolates showed pairing of TIGR4 vex2 with R6 pep27 and had two identical amino acid substitutions, Q441K in vncS and N25D in vex2. These findings indicate that two VTSP strains have emerged independently in Korea, suggesting a prevalence rate of 1.1%. The emergence of VTSP would be a serious threat in Korea, where there are significant rates of penicillin resistance in S. pneumoniae. Monitoring of the prevalence of VTSP and further investigation of the clinical relevance of VTSP are warranted.


A randomized comparative trial of two decision tools for pregnant women with prior cesareans.
Evaluate tools to help pregnant women with prior cesareans make informed decisions about having trials of labor. Randomized comparative trial. A research assistant with a laptop met the women in quiet locations at clinics and at health fairs. Pregnant women (N = 131) who had one prior cesarean and were eligible for vaginal birth after cesarean (VBAC) participated one time between 2005 and 2007. Women were randomized to receive either an evidence-based, interactive decision aid or two evidence-based educational brochures about cesarean delivery and VBAC. Effect on the decision-making process was assessed before and after the interventions. Compared to baseline, women in both groups felt more informed (F = 23.8, p < .001), were more clear about their birth priorities (F = 9.7, p = .002), felt more supported (F = 9.8, p = .002, and overall reported less conflict (F = 18.1, p < 0.001) after receiving either intervention. Women in their third trimesters reported greater clarity around birth priorities after using the interactive decision aid than women given brochures (F = 9.8, p = .003). Although both decision tools significantly reduced conflict around the birth decision compared to baseline, more work is needed to understand which format, the interactive decision aid or paper brochures, are more effective early and late in pregnancy.


Scanning-SAXS of microfluidic flows: nanostructural mapping of soft matter.
The determination of in situ structural information of soft matter under flow is challenging, as it depends on many factors, such as temperature, concentration, confinement, channel geometry, and type of imposed flow. Here, we combine microfluidics and scanning small-angle X-ray scattering (scanning-SAXS) to create a two-dimensional spatially resolved map, which represents quantitatively the variation of molecular properties under flow. As application examples, mappings of confined amyloid fibrils and wormlike micelles under flow into various channel geometries are compared. A simple process to fabricate X-rays resistant chips, based on polyimide and UV-curing resin, is discussed. During experiments, these chips remained in high-energy synchrotron radiation for more than 24 hours, causing constant low background scattering. Thus, sufficient statistics were obtained from sample scattering at exposure times as low as 0.1 s, even with the small scattering volumes in microfluidic channels. Scanning-SAXS of microfluidic flows has many potential applications from biology to fundamental soft matter physics. In general, any fluid which has enough contrast for X-ray scattering can be measured to obtain the dependence of molecular shape, conformation, alignment and size on the flow field. Besides, dynamic processes of soft matter caused by flow, temperature, concentration gradient, and confinement, for example self-assembling, aggregation, mixing, diffusion, and disintegration of macromolecules, can be quantified and visualized on a single image by this mapping technique.


Evolutionary Conservation of ABA Signaling for Stomatal Closure.
Abscisic acid (ABA)-driven stomatal regulation reportedly evolved after the divergence of ferns, during the early evolution of seed plants approximately 360 million years ago. This hypothesis is based on the observation that the stomata of certain fern species are unresponsive to ABA, but exhibit passive hydraulic control. However, ABA-induced stomatal closure was detected in some mosses and lycophytes. Here, we observed that a number of ABA signaling and membrane transporter protein families diversified over the evolutionary history of land plants. The aquatic ferns Azolla filiculoides and Salvinia cucullata have representatives of 23 families of proteins orthologous to those of Arabidopsis (Arabidopsis thaliana) and all other land plant species studied. Phylogenetic analysis of the key ABA signaling proteins indicates an evolutionarily conserved stomatal response to ABA. Moreover, comparative transcriptomic analysis has identified a suite of ABA-responsive genes that differentially expressed in a terrestrial fern species, Polystichum proliferum These genes encode proteins associated with ABA biosynthesis, transport, reception, transcription, signaling, and ion and sugar transport, which fit the general ABA signaling pathway constructed from Arabidopsis and Hordeum vulgare The retention of these key ABA-responsive genes could have had a profound effect on the adaptation of ferns to dry conditions. Furthermore, stomatal assays have shown the primary evidence for ABA-induced closure of stomata in two terrestrial fern species Pproliferum and Nephrolepis exaltata In summary, we report, to our knowledge, new molecular and physiological evidence for the presence of active stomatal control in ferns.


Structure of the SthK carboxy-terminal region reveals a gating mechanism for cyclic nucleotide-modulated ion channels.
Cyclic nucleotide-sensitive ion channels are molecular pores that open in response to cAMP or cGMP, which are universal second messengers. Binding of a cyclic nucleotide to the carboxyterminal cyclic nucleotide binding domain (CNBD) of these channels is thought to cause a conformational change that promotes channel opening. The C-linker domain, which connects the channel pore to this CNBD, plays an important role in coupling ligand binding to channel opening. Current structural insight into this mechanism mainly derives from X-ray crystal structures of the C-linker/CNBD from hyperpolarization-activated cyclic nucleotide-modulated (HCN) channels. However, these structures reveal little to no conformational changes upon comparison of the ligand-bound and unbound form. In this study, we take advantage of a recently identified prokaryote ion channel, SthK, which has functional properties that strongly resemble cyclic nucleotide-gated (CNG) channels and is activated by cAMP, but not by cGMP. We determined X-ray crystal structures of the C-linker/CNBD of SthK in the presence of cAMP or cGMP. We observe that the structure in complex with cGMP, which is an antagonist, is similar to previously determined HCN channel structures. In contrast, the structure in complex with cAMP, which is an agonist, is in a more open conformation. We observe that the CNBD makes an outward swinging movement, which is accompanied by an opening of the C-linker. This conformation mirrors the open gate structures of the Kv1.2 channel or MthK channel, which suggests that the cAMP-bound C-linker/CNBD from SthK represents an activated conformation. These results provide a structural framework for better understanding cyclic nucleotide modulation of ion channels, including HCN and CNG channels.


Thermally induced strain and birefringence calculations for a Nd:YAG rod encapsulated in a solid pump light collector.
Calculations and experimental measurements of the thermally induced strain and birefringence are presented for a diode-pumped Nd:YAG rod that is encapsulated in a prismatic pump light collector. A numerical model is developed to determine the spatiotemporal stress-induced strain distribution across the prism, index-matching fixant, and laser rod, and the birefringence that arises from the stress-induced strain within the laser rod. Calculations of the birefringence are compared with polarscopic measurements and display good agreement. Support for the rod on all sides is provided by the prism and fixant, and the distribution and degree of the stress-induced strain (and birefringence) within the laser rod are therefore influenced by the geometry and composition of the prism and fixant. These strains are thermomechanical in origin and are primarily a function of the elastic modulus of the fixant and the temperature of the system. Such stress-induced strains are additional to those strains that are produced from temperature gradients across the laser rod and result from the laser rod being constrained from expanding. Collectors utilizing index-matching fluid as the encapsulant display the smallest measure of birefringence relating to the temperature gradients in the rod. However, for collectors utilizing solid fixants (with significant elastic modulus), an increase in the birefringence results. In this case collector designs that have the laser rod located in a symmetrically shaped prism are effective in reducing the nonuniform pressures on the sides of the rod and therefore the birefringence.