Graph Drawing by Stochastic Gradient Descent.
A popular method of force-directed graph drawing is multidimensional scaling using graph-theoretic distances as input. We present an algorithm to minimize its energy function, known as stress, by using stochastic gradient descent (SGD) to move a single pair of vertices at a time. Our results show that SGD can reach lower stress levels faster and more consistently than majorization, without needing help from a good initialization. We then show how the unique properties of SGD make it easier to produce constrained layouts than previous approaches. We also show how SGD can be directly applied within the sparse stress approximation of Ortmann et al. [1], making the algorithm scalable up to large graphs.


Evaluation of Candida albicans allergens reactive with specific IgE in asthma and atopic eczema patients.
Candida albicans (C. albicans) produces important allergenic components which can induce allergic reactions in sensitised patients. The purpose of this study was to extract the C. albicans antigens for evaluating the specific anti-Candida IgE in sera of atopic eczema (AE) and asthmatic patients (AS). 95 AE, 85 AS, and 70 non-atopic cases were selected with sequential trials. Candida albicans antigens were prepared and then skin prick test (SPT), ELISA and IgE-immunoblotting tests were performed for all patients. Positive SPT reactions were obtained on 52.6% of AE and 54.1% of AS patients and 4.3% of healthy controls (P < 0.05). Using ELISA, specific anti-C. albicans IgE antibody was detected in 32.6% and 41.2% of patients with AE and AS, respectively. No specific IgE antibody was detected in healthy controls (P < 0.05). In SDS-PAGE, protein bands with molecular weights between 13 and 135 kDa were detected, and some of them reacted with specific IgE in immunoblotting. In AE patients, the most important allergenic components were 25, 34, and 57 kDa protein bands, whereas in AS, 22, 25, and 34 kDa protein bands were observed as major allergens. Candida albicans produces different allergenic components that can induce allergic reactions and may be pathogenetically important in patients with AE and AS.


Modelling host cell availability and the crowding effect in Eimeria infections.
Within-host mathematical models of Eimeria maxima and Eimeria praecox infections of the chicken are presented and used to investigate the role of host cell availability as a possible determinant of the so-called 'crowding effect'; whereby the fecundity of the parasites decreases as infectious dose increases. Assumptions about the number of available host cells, the average lifespan of these cells and the age structure within the host-cell population were made and mathematical models were constructed and combined with experimental data to test whether these conditions could reproduce the crowding effect in the two species. Experimental data demonstrated that crowding during in vivo infections was apparent following very low infectious doses, but none of the models could adequately reproduce crowding at the same doses while maintaining realistic estimates of the dynamics of the enterocyte pool. However, both the size and lifespan of the enterocyte pool were demonstrated to have substantial effects on the fecundity of the infections, particularly at higher doses. These data indicate that host cell availability cannot be solely responsible for the crowding effect. Alternative factors such as the influence of the primary immune response to the parasite may also be explored using within-host models and other applications of these models are discussed.


Homeostatic scaling requires group I mGluR activation mediated by Homer1a.
Homeostatic scaling is a non-Hebbian form of neural plasticity that maintains neuronal excitability and informational content of synaptic arrays in the face of changes of network activity. Here, we demonstrate that homeostatic scaling is dependent on group I metabotropic glutamate receptor activation that is mediated by the immediate early gene Homer1a. Homer1a is transiently upregulated during increases in network activity and evokes agonist-independent signaling of group I mGluRs that scales down the expression of synaptic AMPA receptors. Homer1a effects are dynamic and play a role in the induction of scaling. Similar to mGluR-LTD, Homer1a-dependent scaling involves a reduction of tyrosine phosphorylation of GluA2 (GluR2), but is distinct in that it exploits a unique signaling property of group I mGluR to confer cell-wide, agonist-independent activation of the receptor. These studies reveal an elegant interplay of mechanisms that underlie Hebbian and non-Hebbian plasticity.


Analysis of eye lens-specific genes in congenital hereditary cataracts and microphthalmia of the miniature schnauzer dog.
The congenital hereditary cataracts and microphthalmia in the miniature schnauzer dog are inherited by an autosomal recessive mode. To understand the genetic basis of these diseases, the authors purified and analyzed leukocyte deoxyribonucleic acid (DNA) from affected and normal animals using a candidate gene approach. Because the genes that encode the lens-specific proteins, specifically, alpha, beta, and gamma crystallins and the membrane protein (MP26), are known to maintain the structure and function of the lens, the authors used complimentary DNA (cDNA) fragments that corresponded to the above genes to search for the mutations at their loci in the affected animals. They found no evidence of the gene deletion and rearrangement in any of the five loci. In addition, the hybridizable sequences of the dog DNA to the specific probes for the human chromosome 4 and 18 loci, which are reported to be involved in the abnormality of the human eye, seem to be unaffected. These data support the notion that the hereditary cataracts and microphthalmia in the dog may be associated with genes other than those reported for several animal systems.


NUFIP1 (nuclear FMRP interacting protein 1) is a nucleocytoplasmic shuttling protein associated with active synaptoneurosomes.
Fragile X syndrome, the most common cause of inherited mental retardation, is caused by the absence of FMRP (Fragile X Mental Retardation Protein). FMRP is an RNA binding protein reported to be involved in translational control, notably at postsynaptic sites of protein synthesis as a part of a multiprotein/mRNA complex. One of the FMRP interactors, NUFIP1, is an RNA binding protein with an expression profile matching that of FMRP. We now show that in the nucleus NUFIP1 is localized in the nuclear matrix in RNA-containing structures lying in the proximity of, but not overlapping with, sites of nascent RNA. NUFIP1 is also present in the cytoplasm, where it is associated with ribosomes, similarly to FMRP. In neurons NUFIP1 can be detected in functional synaptoneurosomes, colocalizing with ribosomes. Consistent with its subcellular localization in both nucleus and cytoplasm, we show that NUFIP1 contains a functional CRM1-dependent nuclear export signal and is able to shuttle between these two cellular compartments. These findings suggest the involvement of NUFIP1 in the export and localization of mRNA and, in association with FMRP, in the regulation of local protein synthesis near synapses.


A morphological study of bacterial colonisation of intravenous cannulae.
In an investigation of bacterial colonisation of intravascular cannulae, 16 peripheral venous cannulae of the Venflon variety and 24 chronic haemodialysis cannulae were studied after removal from patients. Studies of colonisation included semi-quantitative microbiological culture and scanning electronmicrographic (SEM) observations. The microbiological findings indicated colonisation of the intravascular portion of the catheter in 4 of 16 Venflon cannulae and 18 of 26 haemodialysis cannulae, largely with skin commensal organisms. The results of the SEM studies were in broad agreement with the microbiological observations. Surface defects on the cannulae were shown to be associated with microbial colonisation which occurred either as isolated colonies or in association with a cellular fibrinous matrix. These observations are illustrated and discussed.


Food Environment and Weight Change: Does Residential Mobility Matter?: The Diabetes Study of Northern California (DISTANCE).
Associations between neighborhood food environment and adult body mass index (BMI; weight (kg)/height (m)2) derived using cross-sectional or longitudinal random-effects models may be biased due to unmeasured confounding and measurement and methodological limitations. In this study, we assessed the within-individual association between change in food environment from 2006 to 2011 and change in BMI among adults with type 2 diabetes using clinical data from the Kaiser Permanente Diabetes Registry collected from 2007 to 2011. Healthy food environment was measured using the kernel density of healthful food venues. Fixed-effects models with a 1-year-lagged BMI were estimated. Separate models were fitted for persons who moved and those who did not. Sensitivity analysis using different lag times and kernel density bandwidths were tested to establish the consistency of findings. On average, patients lost 1 pound (0.45 kg) for each standard-deviation improvement in their food environment. This relationship held for persons who remained in the same location throughout the 5-year study period but not among persons who moved. Proximity to food venues that promote nutritious foods alone may not translate into clinically meaningful diet-related health changes. Community-level policies for improving the food environment need multifaceted strategies to invoke clinically meaningful change in BMI among adult patients with diabetes.


Long-term visual outcome in patients with optic nerve pit and serous retinal detachment of the macula.
Serous detachment of the macula is a well-known complication in patients with an optic nerve pit. Despite the many descriptions of this condition and possible treatment options, the long-term natural history is not well known. The authors identified 15 eyes of 15 consecutive patients seen over 21 years who were diagnosed with a serous detachment of the macula arising from an optic nerve pit. Average length of follow-up was 9 years. Twelve eyes lost three or more lines of vision, two eyes remained unchanged, and only one eye improved. All of the 12 eyes losing three or more lines of vision experienced this decrease within the first 6 months of follow-up. Although only two patients had a visual acuity of 20/200 or less initially, 12 of 15 patients had a visual acuity of 20/200 or less at the time of their last examination. The appearance of the macula at last examination included cystic changes of the neurosensory retina, full-thickness hole formation, retinal pigment epithelial mottling, and lamellar hole formation in the outer retinal layer. The long-term visual prognosis in patients with optic nerve pit and untreated serous retinal detachment of the macula is poor, and visual loss occurs within 6 months of the serous detachment.


How "blind" are double-blind studies?
Psychopharmacological studies usually attempt to eliminate "nonspecific" influences on outcome by double-blind designs. In a randomized, double-blind comparison of alprazolam, imipramine, and placebo, the great majority of panic disorder patients (N = 59) and their physicians were able to rate accurately whether active drug or placebo had been given. Moreover, physicians could distinguish between the two types of active drugs. Inasmuch as correct rating was possible halfway through treatment, concerns about the internal validity of the double-blind strategy arise.


Delay in tuberculosis case detection in Pwani region, Tanzania. A cross sectional study.
Delay in Tuberculosis (TB) case detection may worsen the disease and increase TB transmission. It is also a challenge to the National TB and Leprosy control Program (NTLP). We conducted a cross sectional study in four out of six districts in Pwani region to estimate the extent and factors responsible for delay in TB case detection in Pwani region. Delays were divided into patient, health facility and total delay. We enrolled a total of 226 smear positive TB patients. Out of 226 patient's results were available for 206. The majority (66.5%) of the patients were males. Mean age for males and females were 37.3 and 33.7 years respectively. Mean (SD) total delay was 125.5 (98.5) days (median 90). Out of 206 patients, 79 (38.35%) delayed to seek TB health care. Health facility delay was observed among 121 (58.7%) patients.Risk factors for delay was poor knowledge that chest pain may be a TB symptom (OR = 2.9; 95%CI 1.20- 7.03) and the belief that TB is always associated with HIV/AIDS (OR = 2.7; 95%CI 1.39-5.23). Risk for delay was low among patients who first presented to a government health facility (OR = 0.3; 95%CI 0.12- 0.71) and those presenting with chest pain (OR = 0.2; 95%CI 0.10-0.61). There is a considerable delay in TB case detection in Pwani mainly contributed by patients. Risk factors for delay include misconception about TB/HIV and poor knowledge of TB symptoms.


Biased belief updating and suboptimal choice in foraging decisions.
Deciding which options to engage, and which to forego, requires developing accurate beliefs about the overall distribution of prospects. Here we adapt a classic prey selection task from foraging theory to examine how individuals keep track of an environment's reward rate and adjust choices in response to its fluctuations. Preference shifts were most pronounced when the environment improved compared to when it deteriorated. This is best explained by a trial-by-trial learning model in which participants estimate the reward rate with upward vs. downward changes controlled by separate learning rates. A failure to adjust expectations sufficiently when an environment becomes worse leads to suboptimal choices: options that are valuable given the environmental conditions are rejected in the false expectation that better options will materialize. These findings offer a previously unappreciated parallel in the serial choice setting of observations of asymmetric updating and resulting biased (often overoptimistic) estimates in other domains.


A standardized protocol to reduce cerebrospinal fluid shunt infection: the Hydrocephalus Clinical Research Network Quality Improvement Initiative.
Quality improvement techniques are being implemented in many areas of medicine. In an effort to reduce the ventriculoperitoneal shunt infection rate, a standardized protocol was developed and implemented at 4 centers of the Hydrocephalus Clinical Research Network (HCRN). The protocol was developed sequentially by HCRN members using the current literature and prior institutional experience until consensus was obtained. The protocol was prospectively applied at each HCRN center to all children undergoing a shunt insertion or revision procedure. Infections were defined on the basis of CSF, wound, or pseudocyst cultures; wound breakdown; abdominal pseudocyst; or positive blood cultures in the presence of a ventriculoatrial shunt. Procedures and infections were measured before and after protocol implementation. Twenty-one surgeons at 4 centers performed 1571 procedures between June 1, 2007, and February 28, 2009. The minimum follow-up was 6 months. The Network infection rate decreased from 8.8% prior to the protocol to 5.7% while using the protocol (p = 0.0028, absolute risk reduction 3.15%, relative risk reduction 36%). Three of 4 centers lowered their infection rate. Shunt surgery after external ventricular drainage (with or without prior infection) had the highest infection rate. Overall protocol compliance was 74.5% and improved over the course of the observation period. Based on logistic regression analysis, the use of BioGlide catheters (odds ratio [OR] 1.91, 95% CI 1.19-3.05; p = 0.007) and the use of antiseptic cream by any members of the surgical team (instead of a formal surgical scrub by all members of the surgical team; OR 4.53, 95% CI 1.43-14.41; p = 0.01) were associated with an increased risk of infection. The standardized protocol for shunt surgery significantly reduced shunt infection across the HCRN. Overall protocol compliance was good. The protocol has established a common baseline within the Network, which will facilitate assessment of new treatments. Identification of factors associated with infection will allow further protocol refinement in the future.


Follicle size class contributes to distinct secretion patterns of inhibin isoforms during the rat estrous cycle.
The differential production of inhibins must be exquisitely controlled at the cellular level to ensure the secretion of the appropriate ligand at specific times during the reproductive cycle. The mechanisms underlying inhibin dimer assembly, processing and secretion are not well understood. Here we verify that the secretion of inhibin A and inhibin B from the granulosa cell is discordant during the estrous cycle: discordant production or secretion of the inhibins was not observed during the pregnant mare serum gonadotropin-induced cycle. We correlated the discordant production and secretion of inhibin A and inhibin B into the serum with distinct patterns of inhibin alpha- and beta-subunit colocalization during the cycle in granulosa cells. We determined that the discordant pattern of inhibin A and inhibin B during the rat estrous cycle is due to independent populations of antral follicles making inhibin B (small antral follicles) or inhibin A (large antral follicles).


Extended 2D myotube culture recapitulates postnatal fibre type plasticity.
The traditional problems of performing skeletal muscle cell cultures derived from mammalian or avian species are limited myotube differentiation, and transient myotube persistence which greatly restricts the ability of myotubes to undergo phenotypic maturation. We report here on a major technical breakthrough in the establishment of a simple and effective method of extended porcine myotube cultures (beyond 50 days) in two-dimension (2D) that recapitulates key features of postnatal fibre types. Primary porcine muscle satellite cells (myoblasts) were isolated from the longissimus dorsi of 4 to 6 weeks old pigs for 2D cultures to optimise myotube formation, improve surface adherence and characterise myotube maturation. Over 95 % of isolated cells were myoblasts as evidenced by the expression of Pax3 and Pax7. Our relatively simple approach, based on modifications of existing surface coating reagents (Maxgel), and of proliferation and differentiation (Ultroser G) media, typically achieved by 5 days of differentiation fusion index of around 80 % manifested in an abundance of discrete myosin heavy chain (MyHC) slow and fast myotubes. There was little deterioration in myotube viability over 50 days, and the efficiency of myotube formation was maintained over seven myoblast passages. Regular spontaneous contractions of myotubes were frequently observed throughout culture. Myotubes in extended cultures were able to undergo phenotypic adaptation in response to different culture media, including the adoption of a dominant postnatal phenotype of fast-glycolytic MyHC 2x and 2b expression by about day 20 of differentiation. Furthermore, fast-glycolytic myotubes coincided with enhanced expression of the putative porcine long intergenic non-coding RNA (linc-MYH), which has recently been shown to be a key coordinator of MyHC 2b expression in vivo. Our revised culture protocol allows the efficient differentiation and fusion of porcine myoblasts into myotubes and their prolonged adherence to the culture surface. Furthermore, we are able to recapitulate in 2D the maturation process of myotubes to resemble postnatal fibre types which represent a major technical advance in opening access to the in vitro study of coordinated postnatal muscle gene expression.


Myocardial Extracellular Volume Fraction Adds Prognostic Information Beyond Myocardial Replacement Fibrosis.
Cardiac magnetic resonance techniques permit quantification of the myocardial extracellular volume fraction (ECV), representing a surrogate marker of reactive interstitial fibrosis, and late gadolinium enhancement (LGE), representing replacement fibrosis or scar. ECV and LGE have been independently linked with heart failure (HF) events. In deriving ECV, coronary artery disease type LGE, but not non-coronary artery disease type LGE, has been consistently excluded. We examined the associations between LGE, global ECV derived from myocardial tissue segments free of any detectable scar, and subsequent HF events. Mid short-axis T1 maps were divided into 6 cardiac segments, each classified as LGE absent or present. Global ECV was derived from only segments without LGE. ECV was considered elevated if >30%, the upper 95% bounds of a reference group without known cardiac disease (n=28). Patients were divided into 4 groups by presence of elevated ECV and of any LGE. Subsequent HF hospitalization and any death were ascertained. Their relationship with ECV was examined separately and as a composite with Cox proportional hazard models. Of 1604 serial patients with T1 maps, 1255 were eligible after exclusions and followed over a median 26.3 (interquartile range, 15.9-37.5) months. Patients with elevated ECV had increased risk for death (hazard ratio [HR] 2.45 [95% CI, 1.76-3.41]), HF hospitalization (HR, 2.45 [95% CI, 1.77-3.40]), and a combined end point of both outcomes (HR, 2.46 [95% CI, 1.94-3.14]). After adjustments for covariates including LGE, the relationship persisted for death (HR, 1.82 [95% CI, 1.28-2.59]), hospitalization (HR, 1.60 [95% CI, 1.12-2.27]), and combined end points (HR, 1.73 [95% CI, 1.34-2.24]). ECV measures of diffuse myocardial fibrosis were associated with HF outcomes, despite exclusion of replacement fibrosis segments from their derivation and even among patients without any scar. ECV may have a synergistic role with LGE in HF risk assessment.


Individual and partner risk factors associated with abnormal cervical cytology among women in HIV-discordant relationships.
Individual and sexual partner characteristics may increase the risk of abnormal cervical cytology among women in human immunodeficiency virus (HIV)-discordant relationships. Papanicolaou smears were obtained in a prospective cohort of Kenyan HIV-discordant couples. Of 441 women, 283 (64%) were HIV-infected and 158 (36%) were HIV-uninfected with HIV-infected partners. Overall, 79 (18%) had low-grade and 25 (6%) high-grade cervical abnormalities. Male herpes simplex virus type 2 (HSV-2) seropositivity and lower couple socioeconomic status were associated with cervical abnormalities (p < 0.05). HIV-uninfected women with HIV-infected male sex partners (CD4 > 350 cells/µL) had the lowest prevalence of high-grade cervical lesions. HIV-infected women (CD4 > 350 cells/µL) and HIV-uninfected women with HIV-infected partners (CD4 ≤ 350 cells/µL) were at similar intermediate risk (p > 0.05), and HIV-infected women (CD4 ≤ 350 cells/µL) had significantly higher risk of high-grade cervical abnormalities (p = 0.05). Women in HIV-discordant relationships have high rates of cervical lesions and this may be influenced by couple-level factors, including HIV status and CD4 count of the infected partner.


Regulation of CD45 alternative splicing by heterogeneous ribonucleoprotein, hnRNPLL.
The transition from naïve to activated T cells is marked by alternative splicing of pre-mRNA encoding the transmembrane phosphatase CD45. Using a short hairpin RNA interference screen, we identified heterogeneous ribonucleoprotein L-like (hnRNPLL) as a critical inducible regulator of CD45 alternative splicing. HnRNPLL was up-regulated in stimulated T cells, bound CD45 transcripts, and was both necessary and sufficient for CD45 alternative splicing. Depletion or overexpression of hnRNPLL in B and T cell lines and primary T cells resulted in reciprocal alteration of CD45RA and RO expression. Exon array analysis suggested that hnRNPLL acts as a global regulator of alternative splicing in activated T cells. Induction of hnRNPLL during hematopoietic cell activation and differentiation may allow cells to rapidly shift their transcriptomes to favor proliferation and inhibit cell death.


Spontaneous epiglottic hematoma secondary to supratherapeutic anticoagulation.
Hemorrhage into the soft tissues of the airway represents a potentially life-threatening complication of long-term anticoagulation. We report the case of a chronically anticoagulated 37-year-old male who developed a spontaneous hematoma of the epiglottis secondary to a supra-therapeutic INR. Epiglottic hematoma should be considered in the differential of any anticoagulated patient presenting with upper airway compromise. The airway should be secured in a controlled fashion, and the coagulopathy should be rapidly corrected.


Discrete Polycomb-binding sites in each parasegmental domain of the bithorax complex.
The Polycomb protein of Drosophila melanogaster maintains the segmental expression limits of the homeotic genes in the bithorax complex. Polycomb-binding sites within the bithorax complex were mapped by immunostaining of salivary gland polytene chromosomes. Polycomb bound to four DNA fragments, one in each of four successive parasegmental regulatory regions. These fragments correspond exactly to the ones that can maintain segmentally limited expression of a lacZ reporter gene. Thus, Polycomb acts directly on discrete multiple sites in bithorax regulatory DNA. Constructs combining fragments from different regulatory regions demonstrate that Polycomb-dependent maintenance elements can act on multiple pattern initiation elements, and that maintenance elements can work together. The cooperative action of maintenance elements may motivate the linear order of the bithorax complex.


Analysis of TiO2 Atomic Layer Deposition Surface Chemistry and Evidence of Propene Oligomerization Using Surface-Enhanced Raman Spectroscopy.
Atomic layer deposition (ALD) of TiO2 was performed in tandem with in situ surface-enhanced Raman spectroscopy (SERS) to monitor changes in the transient surface species across multiple ALD cycles. A self-assembled monolayer of 3-mercaptopropionic acid was used as a capture agent to ensure that nucleation of the titanium precursor (titanium tetraisopropoxide [TTIP]) occurs. Comparisons between the Raman spectra of the neat precursor and the SER spectra of the first ALD cycle of TiO2 reveal typical ligand exchange chemistry taking place, with self-limiting behavior and intact isopropoxide ligands. However, subsequent cycles show drastically different chemistry, with no isopropoxide ligands remaining at any point during the second and third cycles. Continuous exposure of either TTIP or isopropyl alcohol after the first cycle shows unlimited chemical vapor deposition (CVD)-type growth. Comparisons with alternative precursors (aluminum isopropoxide, titanium tert-butoxide, and titanium propoxide) and DFT calculations reveal that, for the TTIP precursor, isolated TiO2 sites play a role in the dehydration of off-gassing isopropyl alcohol. The resulting propene then undergoes oligomerization into six-carbon olefins before polymerizing into indistinguishable carbon products that accumulate on the surface. The emergence of the dehydration chemistry is expected to be exclusively the result of these isolated TiO2 sites and, as such, is expected to occur on other surfaces where TiO2 ALD is feasible. This work showcases how seemingly innocuous ALD can evolve into a CVD process when the products can participate in various side reactions with newly made surface sites.


A global perspective of the roles of the pharmacist in the NICU.
To describe pharmacist practice and roles performed in the neonatal intensive care unit (NICU) worldwide and to map these findings along the medicines management pathway (MMP). Quasi-systematic review. Google Scholar, Medline/PubMed and Embase were searched utilising the selected MeSH terms. Thirty sources of information were reviewed. Overall, pharmacist practice in the NICU involves a wide-range of roles, with the most commonly reported involving patient medication chart review, therapeutic drug monitoring and the provision of medication information. Studies highlight that pharmacist contribution to total parenteral nutrition (TPN) regimens and patient medication chart review is beneficial to patient outcomes. Roles beyond the regular scope of practice included involvement in immunisation programmes and research. Most of the data were collected from the USA (13 of 30), followed by the UK (6 of 30) and reports from other countries. The American, British, South African and Australian articles have reported very similar roles, with a pharmacist firmly integrated into the overall structure of the NICU team. The literature identifies that there is insufficient evidence to describe what roles are currently performed in NICUs worldwide. This is due to the lack of recently published articles leading to a large gap in knowledge in understanding what contemporary pharmaceutical services in the NICU comprise. Further research is required to address these gaps in knowledge, and identify the impact of the pharmacist's role on neonatal patient outcomes as well as to determine how to better resource NICUs to access pharmacy services.


Human organoids: a new dimension in cell biology.
Organoids derived from stem cells or tissues in culture can develop into structures that resemble the in vivo anatomy and physiology of intact organs. Human organoid cultures provide the potential to study human development and model disease processes with the same scrutiny and depth of analysis customary for research with nonhuman model organisms. Resembling the complexity of the actual tissue or organ, patient-derived human organoid studies may accelerate medical research, creating new opportunities for tissue engineering and regenerative medicine, generating knowledge and tools for preclinical studies, including drug development and testing. Biologists are drawn to this system as a new "model organism" to study complex disease phenotypes and genetic variability among individuals using patient-derived tissues. The American Society for Cell Biology convened a task force to report on the potential, challenges, and limitations for human organoid research. The task force suggests ways to ease the entry for new researchers into the field and how to facilitate broader use of this new model organism within the research community. This includes guidelines for reproducibility, culturing, sharing of patient materials, patient consent, training, and communication with the public.


Cross-cultural Validation of the 5-Factor Structure of Negative Symptoms in Schizophrenia.
Negative symptoms are currently viewed as having a 2-dimensional structure, with factors reflecting diminished expression (EXP) and motivation and pleasure (MAP). However, several factor-analytic studies suggest that the consensus around a 2-dimensional model is premature. The current study investigated and cross-culturally validated the factorial structure of BNSS-rated negative symptoms across a range of cultures and languages. Participants included individuals diagnosed with a psychotic disorder who had been rated on the Brief Negative Symptom Scale (BNSS) from 5 cross-cultural samples, with a total N = 1691. First, exploratory factor analysis was used to extract up to 6 factors from the data. Next, confirmatory factor analysis evaluated the fit of 5 models: (1) a 1-factor model, 2) a 2-factor model with factors of MAP and EXP, 3) a 3-factor model with inner world, external, and alogia factors; 4) a 5-factor model with separate factors for blunted affect, alogia, anhedonia, avolition, and asociality, and 5) a hierarchical model with 2 second-order factors reflecting EXP and MAP, as well as 5 first-order factors reflecting the 5 aforementioned domains. Models with 4 factors or less were mediocre fits to the data. The 5-factor, 6-factor, and the hierarchical second-order 5-factor models provided excellent fit with an edge to the 5-factor model. The 5-factor structure demonstrated invariance across study samples. Findings support the validity of the 5-factor structure of BNSS-rated negative symptoms across diverse cultures and languages. These findings have important implications for the diagnosis, assessment, and treatment of negative symptoms.


Population diversity of Theileria annulata in Portugal.
The tick-borne protozoan parasite Theileria annulata causes tropical theileriosis, a severe disease of cattle that occurs across the Mediterranean littoral, the Middle East and Southern Asia. In the Mediterranean region, the disease has long been perceived as being a constraint to livestock production in North Africa and Turkey but was believed to have minimal impact in Southern European countries. It has recently been demonstrated that in Southern Portugal the prevalence of T. annulata is approximately 30%. While the population genetics of the parasite and the multiplicity of infection in the bovine host have been studied in a number of countries, no information is currently available on the composition of the parasite population in Southern Europe or its relationship to populations in bordering regions. A parasite genotyping system, based on micro- and mini-satellite amplification, was used to perform genetic analysis of T. annulata populations from T. annulata infected cattle in twelve farms in Southern Portugal. A diversity of genotypes and a high multiplicity of infection were found, suggesting that the parasite possesses a panmictic population in this region. In comparison with genotypes found in Tunisia and Turkey, parasites from Portugal form a genetically distinct group and show lower genetic diversity.


Nest building is a novel method for indexing severity of alcohol withdrawal in mice.
Withdrawal after chronic ethanol (EtOH) affects body temperature, goal-directed behavior and motor function in mice and increases general central nervous system excitability. Nest-building tests have been used to assay these states but to this point have not been employed as measures of EtOH withdrawal severity. We first refined nest-scoring methods using a genetically heterogeneous stock of mice (HS/Npt). Mice were then made physically dependent following three days of chronic EtOH vapor inhalation to produce average blood EtOH concentrations (BECs) of 1.89 mg/mL. EtOH withdrawal affected the progression of nest building over time when mice were tested 2-4 days after removal from three days of chronic exposure to EtOH. In a separate group of mice, chronic EtOH vapor inhalation (BECs 1.84 mg/mL) suppressed nest building over days 1-2 but not days 2-3 of withdrawal. In a following experiment, EtOH withdrawal dose-dependently slowed recovery of nest building for up to 32 h. Finally, we determined that long-lasting nest-building deficits extend to mice undergoing withdrawal from a high dose (4 g/kg) of acute EtOH. Sex differences for nest building were absent following EtOH exposure. In mice naïve to EtOH treatments, male mice had lower pre-test body temperatures and increased nest scores across a two-day testing period compared to females. These results suggest that nest building can be used to assess chronic and acute EtOH withdrawal severity in mice.


Using neutral, selected, and hitchhiker loci to assess connectivity of marine populations in the genomic era.
Estimating the rate of exchange of individuals among populations is a central concern to evolutionary ecology and its applications to conservation and management. For instance, the efficiency of protected areas in sustaining locally endangered populations and ecosystems depends on reserve network connectivity. The population genetics theory offers a powerful framework for estimating dispersal distances and migration rates from molecular data. In the marine realm, however, decades of molecular studies have met limited success in inferring genetic connectivity, due to the frequent lack of spatial genetic structure in species exhibiting high fecundity and dispersal capabilities. This is especially true within biogeographic regions bounded by well-known hotspots of genetic differentiation. Here, we provide an overview of the current methods for estimating genetic connectivity using molecular markers and propose several directions for improving existing approaches using large population genomic datasets. We highlight several issues that limit the effectiveness of methods based on neutral markers when there is virtually no genetic differentiation among samples. We then focus on alternative methods based on markers influenced by selection. Although some of these methodologies are still underexplored, our aim was to stimulate new research to test how broadly they are applicable to nonmodel marine species. We argue that the increased ability to apply the concepts of cline analyses will improve dispersal inferences across physical and ecological barriers that reduce connectivity locally. We finally present how neutral markers hitchhiking with selected loci can also provide information about connectivity patterns within apparently well-mixed biogeographic regions. We contend that one of the most promising applications of population genomics is the use of outlier loci to delineate relevant conservation units and related eco-geographic features across which connectivity can be measured.


Analysis of DNA Double-Strand Breaks and Cytotoxicity after 7 Tesla Magnetic Resonance Imaging of Isolated Human Lymphocytes.
The global use of magnetic resonance imaging (MRI) is constantly growing and the field strengths increasing. Yet, only little data about harmful biological effects caused by MRI exposure are available and published research analyzing the impact of MRI on DNA integrity reported controversial results. This in vitro study aimed to investigate the genotoxic and cytotoxic potential of 7 T ultra-high-field MRI on isolated human peripheral blood mononuclear cells. Hence, unstimulated mononuclear blood cells were exposed to 7 T static magnetic field alone or in combination with maximum permissible imaging gradients and radiofrequency pulses as well as to ionizing radiation during computed tomography and γ-ray exposure. DNA double-strand breaks were quantified by flow cytometry and automated microscopy analysis of immunofluorescence stained γH2AX. Cytotoxicity was studied by CellTiter-Blue viability assay and [3H]-thymidine proliferation assay. Exposure of unstimulated mononuclear blood cells to 7 T static magnetic field alone or combined with varying gradient magnetic fields and pulsed radiofrequency fields did not induce DNA double-strand breaks, whereas irradiation with X- and γ-rays led to a dose-dependent induction of γH2AX foci. The viability assay revealed a time- and dose-dependent decrease in metabolic activity only among samples exposed to γ-radiation. Further, there was no evidence for altered proliferation response after cells were exposed to 7 T MRI or low doses of ionizing radiation (≤ 0.2 Gy). These findings confirm the acceptance of MRI as a safe non-invasive diagnostic imaging tool, but whether MRI can induce other types of DNA lesions or DNA double-strand breaks during altered conditions still needs to be investigated.


STX140 and STX641 cause apoptosis via the intrinsic mitochondrial pathway and down-regulate survivin and XIAP expression in ovarian and prostate cancer cells.
Many anticancer drugs target microtubules and induce apoptosis. However, improved microtubule-targeting drugs, such as STX140 and STX641, are being developed. These compounds induce cell cycle arrest and apoptosis in a variety of tumour cells. The mechanisms that induce apoptosis and the key mediators involved are elucidated in this study. Results demonstrate that STX140 and STX641 depolarise mitochondrial bioenergetics and activate caspase 3/7 in A2780, LNCaP and MCF-7 cancer cells. Furthermore, both compounds cause a significant reduction in the expression of survivin and XIAP. This work details the temporal organisation of apoptosis induced by two microtubule disruptors and highlights the role that the down-regulation of survivin and XIAP may play in this process.


Hypodense alveolar macrophages in patients with diabetes mellitus and active pulmonary tuberculosis.
Alveolar macrophages (AM), a heterogeneous cell population, play a critical role in eliminating mycobacterial infections in collaboration with lymphocytes. Patients with diabetes mellitus (DM) show increased susceptibility to pulmonary tuberculosis (TB) infection. It is still uncertain whether there is a defect in T cell or AM activation in patients with DM against TB infection. To study the difference in activation status of AM and T cells between patients with TB + DM and TB alone. The heterogeneity of AM from 14 patients with TB + DM, 9 with TB alone, 10 normal subjects and 8 DM alone patients, was studied using Percoll density fractionation. The intracellular H2O2 production of AM before and after stimulation with phorbol myristate acetate (PMA) or F-Met-Leu-Phen (FMLP) was assayed by loading cells with 2',7'-dichlorofluorescin (DCFH) and analyzed by flow cytometry. Lymphocytes subsets (CD3, CD4, CD8) and their activation status (CD25) in bronchoalveolar lavage were also measured. The proportion of the least dense AM (< 1,030 g/ml) and the magnitude of DCFH oxidation of AM was higher in TB patients than in normal subjects, regardless of DM. Patients with TB + DM had a significantly lower proportion of the least density AM fraction than TB alone patients, regardless of disease extent. Among TB patients, the proportion of the least dense AM was inversely correlated with the bacterial load on sputum and the disease extent on chest radiograph. Stimulation of AM with PMA or FMLP induced an increase in the hypodense AM subpopulations and enhanced intracellular H2O2 generation in patients with TB + DM and to a similar extent in normal subjects, but not in patients with TB alone. There was no significant difference in CD3 numbers, CD4/CD8 ratio, and CD25+ cells between patients with TB alone and TB + DM. The activation status of AM or T lymphocytes from DM alone patients was not significantly different from those from normal subjects. Hypodense subpopulations of AM increase in active TB patients and are related to the disease severity as well as activation status of AM. AM in TB patients complicated with DM was less activated, and may be contributory to the susceptibility to mycobacterial infection.


A genome-wide association scan on the levels of markers of inflammation in Sardinians reveals associations that underpin its complex regulation.
Identifying the genes that influence levels of pro-inflammatory molecules can help to elucidate the mechanisms underlying this process. We first conducted a two-stage genome-wide association scan (GWAS) for the key inflammatory biomarkers Interleukin-6 (IL-6), the general measure of inflammation erythrocyte sedimentation rate (ESR), monocyte chemotactic protein-1 (MCP-1), and high-sensitivity C-reactive protein (hsCRP) in a large cohort of individuals from the founder population of Sardinia. By analysing 731,213 autosomal or X chromosome SNPs and an additional ∼1.9 million imputed variants in 4,694 individuals, we identified several SNPs associated with the selected quantitative trait loci (QTLs) and replicated all the top signals in an independent sample of 1,392 individuals from the same population. Next, to increase power to detect and resolve associations, we further genotyped the whole cohort (6,145 individuals) for 293,875 variants included on the ImmunoChip and MetaboChip custom arrays. Overall, our combined approach led to the identification of 9 genome-wide significant novel independent signals-5 of which were identified only with the custom arrays-and provided confirmatory evidence for an additional 7. Novel signals include: for IL-6, in the ABO gene (rs657152, p = 2.13×10(-29)); for ESR, at the HBB (rs4910472, p = 2.31×10(-11)) and UCN119B/SPPL3 (rs11829037, p = 8.91×10(-10)) loci; for MCP-1, near its receptor CCR2 (rs17141006, p = 7.53×10(-13)) and in CADM3 (rs3026968, p = 7.63×10(-13)); for hsCRP, within the CRP gene (rs3093077, p = 5.73×10(-21)), near DARC (rs3845624, p = 1.43×10(-10)), UNC119B/SPPL3 (rs11829037, p = 1.50×10(-14)), and ICOSLG/AIRE (rs113459440, p = 1.54×10(-08)) loci. Confirmatory evidence was found for IL-6 in the IL-6R gene (rs4129267); for ESR at CR1 (rs12567990) and TMEM57 (rs10903129); for MCP-1 at DARC (rs12075); and for hsCRP at CRP (rs1205), HNF1A (rs225918), and APOC-I (rs4420638). Our results improve the current knowledge of genetic variants underlying inflammation and provide novel clues for the understanding of the molecular mechanisms regulating this complex process.


Controlling the spin of co atoms on pt(111) by hydrogen adsorption.
We investigate the effect of H adsorption on the magnetic properties of individual Co atoms on Pt(111) with scanning tunneling microscopy. For pristine Co atoms, we detect no inelastic features in the tunnel spectra. Conversely, CoH and CoH2 show a number of low-energy vibrational features in their differential conductance identified by isotope substitution. Only the fcc-adsorbed species present conductance steps of magnetic origin, with a field splitting identifying their effective spin as Seff=2 for CoH and 3/2 for CoH2. The exposure to H2 and desorption through tunnel electrons allow the reversible control of the spin in half-integer steps. Because of the presence of the surface, the hydrogen-induced spin increase is opposite to the spin sequence of CoHn molecules in the gas phase.


Disposition of 1-naphthol in the channel catfish (Ictalurus punctatus).
The pharmacokinetics, tissue distribution, and metabolism of 1-naphthol were examined in the channel catfish (Ictalurus punctatus). Catfish were administered [1-14C]1-naphthol intravascularly at 1, 5, or 25 mg/kg or orally at 1 mg/kg. Plasma data for 1-naphthol were fitted by a three-compartment pharmacokinetic model. There were dose-related changes in the area under the plasma concentration vs. time curve, apparent volume of distribution, and total body clearance after intravascular dosing. After oral dosing, peak plasma concentrations of 1-naphthol occurred at 1 hr; parent compound made up less than 15% of the total radioactivity, and the bioavailability was 32%. Plasma protein binding was 92% and was independent of concentration. 1-Naphthol and metabolites were rapidly eliminated from the tissues after oral dosing; less than 1% of the administered dose remained at 24 hr. Renal excretion was the primary route of elimination of total 14C. Approximately 60% of the oral dose was excreted in the urine within 48 hr. Parent 1-naphthol made up 1% of the urinary 14C. Major metabolites in the urine were sulfate and glucuronide conjugates, which composed 65 and 28% of the total 14C, respectively. Biliary excretion accounted for 7% of the oral dose. The glucuronide conjugate and an unidentified polar metabolite made up the majority of the biliary 14C. The high capacity of channel catfish for conjugative metabolism of 1-naphthol was demonstrated. The dose dependency of pharmacokinetic values could not be explained by saturable metabolism or plasma protein binding.


Dendritic Kv3.3 potassium channels in cerebellar purkinje cells regulate generation and spatial dynamics of dendritic Ca2+ spikes.
Purkinje cell dendrites are excitable structures with intrinsic and synaptic conductances contributing to the generation and propagation of electrical activity. Voltage-gated potassium channel subunit Kv3.3 is expressed in the distal dendrites of Purkinje cells. However, the functional relevance of this dendritic distribution is not understood. Moreover, mutations in Kv3.3 cause movement disorders in mice and cerebellar atrophy and ataxia in humans, emphasizing the importance of understanding the role of these channels. In this study, we explore functional implications of this dendritic channel expression and compare Purkinje cell dendritic excitability in wild-type and Kv3.3 knockout mice. We demonstrate enhanced excitability of Purkinje cell dendrites in Kv3.3 knockout mice, despite normal resting membrane properties. Combined data from local application pharmacology, voltage clamp analysis of ionic currents, and assessment of dendritic Ca(2+) spike threshold in Purkinje cells suggest a role for Kv3.3 channels in opposing Ca(2+) spike initiation. To study the physiological relevance of altered dendritic excitability, we measured [Ca(2+)](i) changes throughout the dendritic tree in response to climbing fiber activation. Ca(2+) signals were specifically enhanced in distal dendrites of Kv3.3 knockout Purkinje cells, suggesting a role for dendritic Kv3.3 channels in regulating propagation of electrical activity and Ca(2+) influx in distal dendrites. These findings characterize unique roles of Kv3.3 channels in dendrites, with implications for synaptic integration, plasticity, and human disease.


Genotyping of common SIRPB1 copy number variant using Paralogue Ratio Test coupled to MALDI-MS quantification.
Copy number variant (CNV) regions have been proven to have a significant impact on gene expression. Some of them have been also found to be associated to different human diseases. CNV genotyping is often prone to error and cross-validation with independent methods is frequently required. The platform of choice depends on whether it is a genome-wide discovery screening or a candidate CNV study, the cohort size and the number of CNVs included in the assay and, finally, the budget available. Here we illustrate a affordable approach to determine the CNV genotype using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and based on the quantitative determination of single nucleotide duplicated mismatches (SNDM) mapping the CNV region and a paralogue genomic region that is used as a two-copy reference. We have genotyped nsv436327, a common CNV mapping SIRPB1 intron 1 that has been associated to human personality behavior. SIRP cluster region was subjected to several ancestral duplication events what makes SIRPB1 CNV genotyping technically challenging. We designed three sets of primer pairs that amplified paralogue regions inside and outside the CNV, containing three SNDMs. Post-PCR extension analyses of sequencing oligonucleotides mapping immediately upstream each SNDM allowed us to quantify using MALDI-MS the proportion of PCR products derived from the CNV region versus the external reference. In contrast to other approaches, setting up this genotyping method requires an affordable investment.


[Professor Hernán Alessandri, M.D.: his legacy for the new generations of physicians].
Professor Alessandri died in 1980. We started our residency in Internal Medicine about 30 years later. Considering the profound changes our society has witnessed, including medical practice, I would like to approach the meaning of his work for our generation. It is not the Father's figure nor his Aura what inspires us today. Neither is his personality nor his shape. His universality comes from his transcendent image as a teacher. Today's teachers live rough times, their social status has changed, their professional requirements have grown exponentially, they have to adapt to social phenomena like the Internet and multiculturalism. Being a teacher nowadays demands to be a multifaceted expert. Things have changed since Professor Alessandri made rounds with his patients. But a deeper look allows us to understand that everything returns to where it started: professional deontology of the teacher, never fading but transcendent. We know that Doctor Alessandri had the natural gift to keep faithful to that code with consistency and perseverance. He excelled with integrity in every aspect including professional betterment, constructive work for his institution, collegiality, a warm relationship with students and a model of social values. Beyond virtues and personal defects he will keep on being the mould in which present teachers should be formed, engraved in their souls and in the subconscious of students that seek to learn.


Direct comparison of the psychometric properties of multiple interview and patient-rated assessments of suicidal ideation and behavior in an adult psychiatric inpatient sample.
Compare the accuracy, agreement, internal consistency, and interrater reliability of 3 interviews to assess suicidal ideation and behavior in accordance with US Food and Drug Administration guidance about reporting categories. Adults admitted to a psychiatric inpatient unit (N = 199) completed 3 assessments of past month and lifetime suicidal ideation and behavior-the Columbia Suicide Severity Rating Scale (C-SSRS), the Suicide Tracking Scale (STS), and the Sheehan Suicidality Tracking Scale (S-STS)-in randomized, counterbalanced order. "Missing gold standard" latent class analyses defined categories for ideation and behavior. Analyses also evaluated the S-STS mapping to C-SSRS categories. Three trained judges re-rated 89 randomly selected interview videotapes. Cohen κ, the primary outcome measure, quantified agreement above chance. Data were collected between November 2011 and June 2013. All 3 assessments showed excellent accuracy for suicidal ideation (κ = 0.72 to 1.00) and attempts (κ = 0.82 to 0.95) calibrated against latent classes. Interrater agreement ranged from κ = 0.52 to 1.00. Interrater agreement about more granular C-SSRS categories varied more widely (κ = 0.48 to 1.00), and the C-SSRS and S-STS assigned significantly different numbers of cases to many categories. Cronbach α was < 0.55 for the C-SSRS ideation and between 0.78 and 0.92 for the other scales. All 3 assessments showed good accuracy for broad categories of suicidal ideation and behavior. More granular, specific categories usually were rated reliably, but the C-SSRS and S-STS differed significantly in regard to which patients were assigned to these subcategories. Using any of these interviews would improve reliability over unstructured assessment in evaluating suicidal ideation and behavior. Clinical predictive validity of these interviews, and particularly the more granular categories, remains to be shown.


Central osmoregulatory influences on thermoregulation.
1. Many mammals maintain a constant core body temperature in the face of a heat load by using evaporative cooling responses, such as sweating, panting and spreading of saliva. These cooling mechanisms incur a body fluid deficit if the fluid lost as sweat, saliva or respiratory moisture is not replaced by the ingestion of water; body fluid hypertonicity and hypovolaemia result. 2. Evidence in several mammals shows that, as they become dehydrated, evaporative cooling mechanisms such as sweating and panting are inhibited so that further fluid loss from the body is reduced. As a result, core temperature in the dehydrated animal is maintained at a higher than normal level. 3. Increasing the osmotic pressure of plasma has an inhibitory effect on panting and sweating in mammals. It has been proposed that osmoreceptors mediate these inhibitory influences of plasma hypertonicity on sweating and panting. 4. The suppression of panting in dehydrated sheep is mediated by cerebral osmoreceptors that are probably located in the lamina terminalis. We speculate that osmoreceptors in the lamina terminalis may also influence thermoregulatory sweating. 5. When dehydrated animals drink water, sweating and panting resume rapidly before water has been absorbed from the gut. It is likely that the act of drinking initiates a reflex that can override the osmoreceptor inhibition of panting, resulting in core temperature falling back quickly to a normal level.


Genetic modifiers of Leprfa associated with variability in insulin production and susceptibility to NIDDM.
In an attempt to identify the genetic basis for susceptibility to non-insulin-dependent diabetes mellitus within the context of obesity, we generated 401 genetically obese Leprfa/Leprfa F2 WKY13M intercross rats that demonstrated wide variation in multiple phenotypic measures related to diabetes, including plasma glucose concentration, percentage of glycosylated hemoglobin, plasma insulin concentration, and pancreatic islet morphology. Using selective genotyping genome scanning approaches, we have identified three quantitative trait loci (QTLs) on Chr. 1 (LOD 7.1 for pancreatic morpholology), Chr. 12 (LOD 5.1 for body mass index and LOD 3.4 for plasma glucose concentration), and Chr. 16 (P < 0.001 for genotype effect on plasma glucose concentration). The obese F2 progeny demonstrated sexual dimorphism for these traits, with increased diabetes susceptibility in the males appearing at approximately 6 weeks of age, as sexual maturation occurred. For each of the QTLs, the linked phenotypes demonstrated sexual dimorphism (more severe affection in males). The QTL on Chr. 1 maps to a region vicinal to that previously linked to adiposity in studies of diabetes susceptibility in the nonobese Goto-Kakizaki rat, which is genetically closely related to the Wistar counterstrain we employed. Several candidate genes, including tubby (tub), multigenic obesity 1 (Mob1), adult obesity and diabetes (Ad), and insulin-like growth factor-2 (Igf2), map to murine regions homologous to the QTL region identified on rat Chr. 1.


Characteristics and risk factors in juvenile sexual offenders.
It is estimated that juvenile sexual offences constitute about 7% of the total annual rate of sexual offences in Spain. Nevertheless, research on Spanish juvenile sex offenders (JSO) is virtually non-existent. This paper analyzes the risk factors related to sexual violence committed by adolescents. The participants were 73 adolescents (M = 15.68 years, SD = 1.12) aged between 14 and 18, who were serving a sentence for committing a sexual offence in various Spanish Autonomous Regions. In this descriptive study multiple methods were used to collect the data: court records, self-reports, along with an interview with the JSO and with the professionals involved. Risk factors related to family history, certain personality characteristics, and the development of "inadequate sexualisation" (96% of cases) were analyzed. This latter variable was mainly related to an early onset of pornography consumption (70%), to a sexualized family environment (26%), and to sexual victimization during childhood (22%). These results are consistent with international research on juvenile sex offending, so we can conclude that the process of development of sexualization from infancy onwards should be deeply examined with regard to sexual violence.


Selegiline in the treatment of attention deficit hyperactivity disorder in children: a double blind and randomized trial.
Attention deficit hyperactivity disorder (ADHD) is a common disorder of childhood that affects 3% to 6% of school-age children. Conventional stimulant medications are recognized by both specialists and parents as useful symptomatic treatment. Nevertheless, approximately 30% of ADHD children treated with them do not respond adequately or cannot tolerate the associated adverse effects. Such difficulties highlight the need for alternative safe and effective medications in the treatment of this disorder. Selegiline is a type B monoamine oxidase inhibitor (MAOI) that is metabolized to amphetamine and methamphetamine stimulant compounds that may be useful in the treatment of ADHD. The authors undertook this study to further evaluate, under double-blind and controlled conditions, the efficacy of selegiline for ADHD in children. A total of 28 children with ADHD as defined by DSM IV were randomized to selegiline or methylphenidate dosed on an age and weight-adjusted basis at selegiline 5 mg/day (under 5 years) and 10 mg/day (over 5 years) (Group 1) and methylphenidate 1 mg/kg/day (Group 2) for a 4-week double-blind clinical trial. The principal measure of the outcome was the Teacher and Parent ADHD Rating Scale. Patients were assessed by a child psychiatrist at baseline, 14 and 28 days after the medication started. No significant differences were observed between the two protocols on the Parent and Teacher Rating Scale scores. Although the number of dropouts in the methylphenidate group was higher than in the selegiline group, there was no significant difference between the two protocols in terms of the dropouts. Decreased appetite, difficulty falling asleep and headaches were observed more in the methylphenidate group. The results of this study must be considered preliminary, but they do suggest that selegiline may be beneficial in the treatment of ADHD. In addition, a tolerable side effect profile may be considered as one of the advantages of selegiline in the treatment of ADHD.


Chiropractic Management of Infantile Torticollis With Associated Abnormal Fixation of One Eye: A Case Report.
The purpose of this case report is to describe the chiropractic management of a child with abnormal fixation of one eye and torticollis. A mother presented with a concern regarding her 23-month-old son who had a history of torticollis and an abnormal fixation of the right eye. She noticed the head tilt when he was about 7 months old and abnormal alignment of the right eye when the boy was 18 months old. At 15 months when he took his first steps, his head tilt became worse. At 21 months old, a neurological and orthopedic examination at the regional university children`s hospital ruled out presence of a tumor of the cervical spine or posterior fossa. The orthopedist sent the baby for chiropractic evaluation and treatment. Chiropractic exam found decreased active and passive range of motion in the cervical spine and no evidence of mass or contracture of the sternocleidomastoid muscle. Segmental palpation showed a decreased joint play and pain reaction at level C1/C2 on the right. The chiropractic treatment consisted of spinal manipulative therapy of the cervical spine in addition to massage and stretching of the neck muscles. Within a period of 4 weeks (3 chiropractic treatments) the torticollis was nearly resolved and the abnormal fixation of the right eye was no longer apparent. No relapse of the symptomatology was observed at a follow-up consultation at 26 months. The patient responded favorably to chiropractic care, showing a possible mechanical spinal cause for his torticollis and for the secondarily developed abnormal fixation of the right eye.


High-dose human lymphoblastoid interferon in metastatic colorectal cancer: clinical results and modification of biological responses.
A total of 22 patients with advanced measurable colorectal carcinoma were treated with human lymphoblastoid interferon, 15 X 10(6) U/m2 im 3 times a week, in a trial designed to evaluate therapeutic activity, toxic effects, and biological response modification. One partial response (4.5% response rate) was observed which lasted 4 months. Sixty-eight percent of the patients required dose reduction for excessive toxicity, primarily constitutional symptoms. One patient developed phenobarbital toxicity, a previously undescribed side effect thought to be related to interferon-induced depression of hepatic microsomal enzymes required for drug metabolism. Treatment was associated with an increase in peripheral blood natural killer (NK) cell activity and the activity of an interferon-induced enzyme, 2'-5' oligoadenylate synthetase. The increase in NK cell activity was observed only in patients whose pretreatment NK cell activity was below normal. No induction of serum factors inducing differentiation of myeloid leukemic cell lines was documented. We conclude that human lymphoblastoid interferon, at the dose and schedule tested, has minimal antitumor activity as a single agent in advanced colorectal cancer and induces unacceptable toxicity in the majority of such patients. Recent literature suggesting a possible role for interferon alpha in combination with other drugs in the treatment of colorectal cancer is discussed.


Diagnosis and Management of Infantile Hemangioma.
Infantile hemangiomas (IHs) are the most common tumors of childhood. Unlike other tumors, they have the unique ability to involute after proliferation, often leading primary care providers to assume they will resolve without intervention or consequence. Unfortunately, a subset of IHs rapidly develop complications, resulting in pain, functional impairment, or permanent disfigurement. As a result, the primary clinician has the task of determining which lesions require early consultation with a specialist. Although several recent reviews have been published, this clinical report is the first based on input from individuals representing the many specialties involved in the treatment of IH. Its purpose is to update the pediatric community regarding recent discoveries in IH pathogenesis, treatment, and clinical associations and to provide a basis for clinical decision-making in the management of IH.


Influence of workplace environment, working conditions and health status of taxi drivers on vehicle collisions or near-miss events.
Sudden illness while driving has been identified as a major cause of vehicle collisions, particularly among taxi drivers. However, no previous studies have examined the factors contributing to the occurrence of health-related vehicle collisions. The current study aimed to identify the risk factors causing health-related vehicle collisions among taxi drivers, and to inform preventative interventions. A cross-sectional survey involving a written anonymous questionnaire was conducted for company-employed taxi drivers in Chiba Prefecture, Japan. A total of 1,739 drivers returned questionnaires. Drivers were classified into those who had experienced collisions or near-miss incidents caused by health problems (Event group), and those who had not (Non-event group). According to a multivariable logistic regression analysis, being unable to take vacation time (odds ratio [OR] 4.59, 95% confidence interval [CI] 2.2-9.58), having chronic diseases (OR 1.52, 95% CI 1.02-2.27), taking insufficient vacation time (OR 1.81, 95% CI 1.19-2.79), and difficulty reporting poor health conditions (OR 2.29, 95% CI 1.36-3.87) were significant factors for the likelihood of an event. Support for drivers to maintain well-balanced control of illnesses and improvement of working environments could prevent health-related events while driving.


The pharmacists' ability to use pressurized metered-dose inhalers with a spacer device and factors affecting it.
Objective: The aim of this study is to evaluate the pharmacists' ability to use pMDIs with a spacer device and the factors that affect this ability.Method: Face to face interviews were conducted with the pharmacists. A nine item questionnaire was completed and the checklist for how to use pMDIs with a spacer device was filled out.Results: A total of 307 pharmacists voluntarily participated in this study. Fifty-six (18.2%) of the pharmacists stated that they did not know how to use pMDIs with a spacer device. These pharmacists were excluded and remaining 251 pharmacists included in the study. Only 100 (39.8%) pharmacists demonstrated all of the inhaler spacer device usage steps correctly. The step in which pharmacists made the most mistakes was "take 5-6 deep and slow breaths, hold for 10 s and slow breaths." Those pharmacists who were more likely to correctly use pMDIs with a spacer device were younger (p = 0.023), had dispensed more asthma medications per day (p < 0.001), had dispensed more asthma medications per day for patients younger than six years of age (p = 0.016), and sold inhaler spacer devices at their pharmacy (p = 0.042).Conclusion: Approximately one third of the pharmacists in the current study were able to correctly demonstrate all of the steps for proper usage of pMDIs with a spacer device, which indicates that pharmacists should be included in the training program and be provided continuous training on the use of pMDIs with a spacer device.


Use of ChAd3-EBO-Z Ebola virus vaccine in Malian and US adults, and boosting of Malian adults with MVA-BN-Filo: a phase 1, single-blind, randomised trial, a phase 1b, open-label and double-blind, dose-escalation trial, and a nested, randomised, double-blind, placebo-controlled trial.
The 2014 west African Zaire Ebola virus epidemic prompted worldwide partners to accelerate clinical development of replication-defective chimpanzee adenovirus 3 vector vaccine expressing Zaire Ebola virus glycoprotein (ChAd3-EBO-Z). We aimed to investigate the safety, tolerability, and immunogenicity of ChAd3-EBO-Z in Malian and US adults, and assess the effect of boosting of Malians with modified vaccinia Ankara expressing Zaire Ebola virus glycoprotein and other filovirus antigens (MVA-BN-Filo). In the phase 1, single-blind, randomised trial of ChAd3-EBO-Z in the USA, we recruited adults aged 18-65 years from the University of Maryland medical community and the Baltimore community. In the phase 1b, open-label and double-blind, dose-escalation trial of ChAd3-EBO-Z in Mali, we recruited adults 18-50 years of age from six hospitals and health centres in Bamako (Mali), some of whom were also eligible for a nested, randomised, double-blind, placebo-controlled trial of MVA-BN-Filo. For randomised segments of the Malian trial and for the US trial, we randomly allocated participants (1:1; block size of six [Malian] or four [US]; ARB produced computer-generated randomisation lists; clinical staff did randomisation) to different single doses of intramuscular immunisation with ChAd3-EBO-Z: Malians received 1 × 10(10) viral particle units (pu), 2·5 × 10(10) pu, 5 × 10(10) pu, or 1 × 10(11) pu; US participants received 1 × 10(10) pu or 1 × 10(11) pu. We randomly allocated Malians in the nested trial (1:1) to receive a single dose of 2 × 10(8) plaque-forming units of MVA-BN-Filo or saline placebo. In the double-blind segments of the Malian trial, investigators, clinical staff, participants, and immunology laboratory staff were masked, but the study pharmacist (MK), vaccine administrator, and study statistician (ARB) were unmasked. In the US trial, investigators were not masked, but participants were. Analyses were per protocol. The primary outcome was safety, measured with occurrence of adverse events for 7 days after vaccination. Both trials are registered with ClinicalTrials.gov, numbers NCT02231866 (US) and NCT02267109 (Malian). Between Oct 8, 2014, and Feb 16, 2015, we randomly allocated 91 participants in Mali (ten [11%] to 1 × 10(10) pu, 35 [38%] to 2·5 × 10(10) pu, 35 [38%] to 5 × 10(10) pu, and 11 [12%] to 1 × 10(11) pu) and 20 in the USA (ten [50%] to 1 × 10(10) pu and ten [50%] to 1 × 10(11) pu), and boosted 52 Malians with MVA-BN-Filo (27 [52%]) or saline (25 [48%]). We identified no safety concerns with either vaccine: seven (8%) of 91 participants in Mali (five [5%] received 5 × 10(10) and two [2%] received 1 × 10(11) pu) and four (20%) of 20 in the USA (all received 1 × 10(11) pu) given ChAd3-EBO-Z had fever lasting for less than 24 h, and 15 (56%) of 27 Malians boosted with MVA-BN-Filo had injection-site pain or tenderness. 1 × 10(11) pu single-dose ChAd3-EBO-Z could suffice for phase 3 efficacy trials of ring-vaccination containment needing short-term, high-level protection to interrupt transmission. MVA-BN-Filo boosting, although a complex regimen, could confer long-lived protection if needed (eg, for health-care workers). Wellcome Trust, Medical Research Council UK, Department for International Development UK, National Cancer Institute, Frederick National Laboratory for Cancer Research, Federal Funds from National Institute of Allergy and Infectious Diseases.


Heparin-bonded central venous catheters do not reduce thrombosis in infants with congenital heart disease: a blinded randomized, controlled trial.
Infants with congenital heart disease who require central venous lines are at increased risk of thrombosis. Heparin-bonded catheters provide protection from thrombotic events in some children. However, heparin-bonded catheters may not be as effective in infants </=1 year old because of other potential risk factors (smaller vessel size, longer duration of catheter use). No studies have assessed the benefit of heparin-bonded catheters in such specific high-risk populations. The objective of this study was to assess the efficacy of heparin-bonded catheters for preventing thrombosis in infants aged </=1 year with congenital heart disease. This study was designed as a randomized, controlled, blinded single-center trial. Infants </=1 year old with congenital heart disease requiring a central venous line for clinical care were randomly assigned to receive either a heparin-bonded catheter or a standard non-heparin-bonded catheter. Catheters were visually indistinguishable. The primary outcome was incidences of both silent and clinically noticeable thrombosis confirmed by ultrasound. Ultrasounds were reviewed by a blinded central adjudication committee. Interim analysis was performed after enrollment of 97 patients. Eighty-seven patients were evaluable (41 of the patients were female). Thrombotic events occurred in 17 (42.5%) of 40 patients in the non-heparin-bonded catheter group and in 21 (44.7%) of 47 patients in the heparin-bonded catheters group. The study was stopped when the interim analysis showed convincing evidence for no difference between groups over the alternative hypothesis of 50% risk reduction. Infants with congenital heart disease are at significant risk of both silent and clinically identified thrombosis. There seems to be no advantage in using heparin-bonded catheters in infants </=1 year of age.


Sex fantasies revisited: an expansion and further clarification of variables affecting sex fantasy production.
Findings of prior research which indicated both external factors (experimental response cues) and internal factors (sex guilt) influencing subjects' writing of their sexual fantasies were expanded. University males (N = 88) and females (N = 122) were studied. They read sex fantasy examples varying in conventionality of the sexual activity and the relationship context before reporting their own fantasies. Sex guilt level of all subjects was measured as well as their subjective responses to writing the fantasies. The data were analyzed to determine whether varying response cues along certain dimensions could influence subjects to write parallel fantasies. Sex guilt levels and sex of the subject were the best predictors of the length, explicitness, and variety of content of subjects' reported fantasies. Males wrote longer, more explicit and varied fantasies but were affected similarly by sex guilt levels as females. High sex guilt subjects in general produced more restricted content and shorter fantasies. They also experienced more embarrassment and less arousal. Females were more likely to have high levels of guilt about sex than males. The varying dimensions of the fantasy examples had little effect on the types of fantasy material revealed and therefore the erotic/explicit aspect of the examples found in the prior research seems to be the factor that produces effects on fantasy production. Comparing the two studies, females may respond with longer, more explicit fantasies when given erotic but more traditional sexual examples.


Disproportionate intraventricular viscous energy loss in Fontan patients: analysis by 4D flow MRI.
To non-invasively assess intraventricular viscous energy loss (EL) and proportionality to kinetic energy (KE) in Fontan patients using 4D flow magnetic resonance imaging (MRI) and compare to healthy controls. Thirty Fontan patients and 15 controls underwent 4D flow MRI. Ventricular EL was computed and normalized by end-diastolic volume (EDV). EL over the cardiac cycle (ELcycle) relative to the average kinetic energy (KEcycle) was quantified using a dimensionless index, ELindex (ELcycle/KEcycle). EL was significantly elevated in Fontan patients compared to controls (ELcycle/EDV: 3.7 vs. 2.6 J/m3, P = 0.001). Fontan patients with inflow of blood entering a ventricle, which leaves that ventricle through a ventricular septal defect to the aorta, defined as discordant inflow-to-outflow showed significantly higher EL compared to patients with inflow of blood into the same ventricle from which the blood is ejected into the aorta, defined as concordant inflow-to-outflow (ELcycle/EDV: 6.8 vs. 3.2 J/m3, P = 0.001). EL was not significantly different in Fontan patients with a systemic left vs. a systemic right ventricle (ELcycle/EDV: 3.5 vs. 3.6 J/m3, P = 0.91). ELindex was significantly elevated in Fontan patients compared to controls (ELindex: 0.4 vs. 0.2, P < 0.001). Highest ELindex was found in Fontan patients with discordant inflow-to-outflow compared to concordant inflow-to-outflow (0.5 vs. 0.3, P = 0.01). Fontan patients show disproportionate intraventricular EL relative to KE. Intraventricular EL is elevated in these patients compared to healthy controls, and highest EL is found in patients with discordant inflow-to-outflow.


What do we perceive from motion pictures? A computational account.
Cinema viewed from a location other than a canonical viewing point (CVP) presents distortions to the viewer in both its static and its dynamic aspects. Past works have investigated mainly the static aspect of this problem and attempted to explain why viewers still seem to perceive the scene very well. The dynamic aspect of depth perception, which is known as structure from motion, and its possible distortion, have not been well investigated. We derive the dynamic depth cues perceived by the viewer and use the so-called isodistortion framework to understand its distortion. The result is that viewers seated at a reasonably central position experience a shift in the intrinsic parameters of their visual systems. Despite this shift, the key properties of the perceived depths remain largely the same, being determined in the main by the accuracy to which extrinsic motion parameters can be recovered. For a viewer seated at a noncentral position and watching the movie screen at a slant angle, the view is related to the view at the CVP by a homography, resulting in various aberrations such as noncentral projection.


Surgical treatment of carcinoma of the gastric cardia.
The ideal surgical treatment for adenocarcinoma of the gastric cardia is still controversial. In 189 consecutive patients who underwent resection, 127 esophagogastric resections and 62 total gastrectomies plus esophageal resection were performed. Short- and long-term results of the two surgical procedures were compared in order to define the specific indications for each. Abdominal nodes were metastatic in 73.5% of the cases, and mediastinal nodes in 29.1% of the patients who were also approached through the thorax. Neoplastic permeation of the esophageal resection margin occurred in 3.2% of the patients. No positive resection margins were found in the cases in whom 10 or more cm. of uninvolved esophagus were resected. The superiority of the laparotomy and right thoracotomy approach was thus evident in terms of oncologic radicality. Anastomotic leakage occurred in 8.7% of esophagogastric resection, and in 6.5% of total gastrectomy plus esophageal resection, patients. No correlation between the stage of the tumor or the neoplastic permeation of the section margin and the incidence of anastomotic leakage was found. Operative mortality was 3.9% after esophagogastric resection, and 6.5% after total gastrectomy plus esophageal resection; this may suggest that esophagogastric resection is the procedure of choice in poor risk and elderly patients. After curative resection, locoregional or systemic neoplastic recurrence was observed in 15.0% of the cases.(ABSTRACT TRUNCATED AT 250 WORDS)


Pharmacogenetics of intestinal absorption.
The small intestine is the primary site of absorption for many drugs administered orally and so is the target tissue for pharmacotherapeutic strategies to control the oral absorption of drugs. Drug transporters, including the ATP-binding cassette (ABC) superfamily and the solute carrier (SLC) superfamily, have been considered to play a physiological role in regulating the absorption of xenobiotics, and variations in their expression level and function in the small intestine cause intra- and inter-individual variation in the oral absorption of drugs. Recent advances in molecular biology have suggested that genetic polymorphisms are associated with the expression level and function, and thereby inter-individual variation. In this review, the pharmacogenetics of these transporters is summarized, and their future significance in the clinical setting is discussed.


Evaluation of early postoperative results after bicaval versus standard cardiac transplantation and review of the literature.
Previous studies have been inconsistent in defining a clinical benefit to the bicaval cardiac transplantation technique relative to the standard technique, and many major centers have not adopted this newer approach. The purpose of this study was to determine whether clinically significant benefits support utilization of the bicaval technique. Sixty-eight consecutive adult patients undergoing a standard cardiac transplant were compared with 75 consecutive patients who underwent the bicaval technique during the period from 1991 to 1999. Etiology, recipient sex, recipient age, donor age, and pulmonary vascular resistance were similar between the two groups. Cardiac index at 24 hours after operation was increased for the bicaval group relative to the standard group (3.15 +/- 0.7 vs 2.7 +/- 0.5 L/min/m(2), P <. 05). Inotropic requirements were significantly less, and there was significantly less tricuspid regurgitation in the bicaval group relative to the standard group. In addition, the bicaval group more frequently had a nonpaced normal sinus rhythm at 24 hours after operation (73.9% vs 50.7% [standard group], P =.025) and had fewer postoperative arrhythmias (29.3% vs 47.7% [standard group], P <.01). Finally, although mortality was similar for the two groups, length of postoperative hospitalization was longer for the standard group relative to the bicaval group (12.1 +/- 11 vs 20.4 +/- 12 days, P <. 001). Review of the literature identified reduced tricuspid regurgitation and improved rhythm as consistent benefits of the bicaval technique. This review demonstrates a clinical benefit during the early postoperative period with bicaval cardiac transplantation (relative to standard) and encourages further utilization of this technique.


Observational study on preoperative surgical field disinfection: povidone-iodine and chlorhexidine-alcohol.
Surgical site infection (SSI) rate is reported to range around 16%. Preoperative skin disinfection is keystone for SSI reduction. Chlorhexidine-alcohol has been reported to be more effective than Povidone-iodine (PVI). However, in many countries established habits and the inferior costs of PVI restrain the employment of chlorhexidine disinfection kits (ChloraPrep®) for the preparation of the surgical field. The costs of surgical field preparation in clean-contaminated surgery utilizing PVI (Betadine) and chlorhexidine alcohol and the evaluation of surgeon compliance and satisfaction, were studied by a observational study on 50 surgical operations in which surgical field was prepared with PVI checking established guidelines, and on 50 surgical operations in which chlorhexidine-alcohol (ChloraPrep) was employed. The use of auxiliary material was tabulated as well as the timing of the phases of disinfection and the surgeon's opinions. The use of auxiliary material (gloves, gauzes, paper towels, surgical instruments, small swabs for umbilical cleaning) is associated with the type of disinfectant, with major use of auxiliary materials recorded in PVI disinfection. PVI disinfection does not follow stringent guidelines, in particular waiting for the disinfectant to dry. PVI guidelines are more demanding than those relative to ChloraPrep. The time necessary for the preparation of the field is significantly longer for PVI. Auxiliary material and guideline compliance must be taken into account when calculating costs; the former are direct costs (even though marginal) and the latter can determine major infective risk. Chlorhexidine in kits is easier and faster to use than PVI, requires less auxiliary material and has been shown previously to reduce SSI in clean contaminated surgery.


Response of serum macrophage migration inhibitory factor levels to stimulation or suppression of the hypothalamo-pituitary-adrenal axis in normal subjects and patients with Cushing's disease.
Macrophage migration inhibitory factor (MIF) is a proinflammatory pituitary and immune cell cytokine and a critical mediator of septic shock. It has been reported that MIF is secreted in parallel with ACTH from the pituitary in response to stress or inflammatory stimuli. MIF release from immune cells is also induced rather than inhibited by glucocorticoids. It has therefore been suggested that MIF may be a novel counterregulatory hormone of glucocorticoid action that acts both as a paracrine and endocrine modulator of host responses. We have measured circulating MIF levels, using a human MIF ELISA, in normal subjects and patients under numerous pathophysiological conditions. Serum MIF was measured in normal subjects who underwent stimulation of the hypothalamo-pituitary-adrenal axis with an insulin tolerance test (n = 8), a CRH-stimulation test (n = 5), a short synacthen test (n = 5), and following a low-dose dexamethasone suppression test (n = 6). We also sampled from a peripheral vein and both inferior petrosal sinuses before and after CRH stimulation in four patients with a histologically proven diagnosis of Cushing's disease. Immunostaining of the pituitary tumors for MIF was also performed. In normal subjects serum MIF levels did not rise in parallel with cortisol during the insulin tolerance or CRH test or after administration of synthetic ACTH. In all subjects cortisol levels became undetectable after the low-dose dexamethasone suppression test, and no consistent change was observed in serum MIF levels during the test. In patients with Cushing's disease, there was no basal central-to-peripheral gradient in MIF, and no consistent changes occurred in serum MIF levels in either the left or right inferior petrosal sinus after CRH stimulation; however, immunostaining of the surgically removed pituitary tumors from the same patients showed strong staining for both ACTH and MIF. These results show that in humans acute modulation of the hypothalamo-pituitary-adrenal axis does not significantly alter circulating MIF levels. In addition, ACTH-secreting pituitary tumors that express MIF do not release MIF either spontaneously or in response to CRH stimulation, and there is no gradient for MIF in the venous drainage of the pituitary. Our study suggests that the pituitary gland is not the major contributor to circulating MIF; an autocrine or paracrine role for pituitary-derived MIF is more likely.


The effects of excipients and particle engineering on the biophysical stability and aerosol performance of parathyroid hormone (1-34) prepared as a dry powder for inhalation.
Pulmonary delivery of therapeutic peptides and proteins has many advantages including high relative bioavailability, rapid systemic absorption and onset of action and a non-invasive mode of administration which improves patient compliance. In this study, we investigated the effect of spray-drying (SD) and spray freeze-drying processes on the stability and aerosol performance of parathyroid hormone (PTH) (1-34) microparticles. In this study, the stabilisation effect of trehalose (a non-reducing sugar) and Brij 97 (a non-ionic surfactant) on spray-dried PTH particles was assessed using analytical techniques including circular dichroism (CD), fluorescence spectroscopy, modulated differential scanning calorimetry and an in vitro bioactivity assay. Physical characterisation also included electron microscopy, tap density measurement and laser light diffraction. The aerosol aerodynamic performance of the formulations was assessed using the Andersen cascade impactor. Based on these studies, a formulation for spray freeze-drying was selected and the effects of the two particle engineering techniques on the biophysical stability and aerosol performance of the resulting powders was determined. CD, fluorescence spectroscopy and bioactivity data suggest that trehalose when used alone as a stabilising excipient produces a superior stabilising effect than when used in combination with a non-ionic surfactant. This highlights the utility of CD and fluorescence spectroscopy studies for the prediction of protein bioactivity post-processing. Therefore, a method and formulation suitable for the preparation of PTH as a dry powder was developed based on spray-drying PTH with trehalose as a stabiliser with the bioactivity of SD PTH containing trehalose being equivalent to that of unprocessed PTH.


Regulation of early mRNA synthesis after bacteriophage T4 infection of Escherichia coli.
Regulation of T4-specific mRNA synthesis was studied during leucine starvation of a leucine-requiring stringent Escherichia coli B strain. This was done by imposing starvation prior to T4 infection and then letting RNA synthesis proceed for different time periods. Rifampin or streptolydigin was added to stop further RNA synthesis, and protein synthesis was restored by addition of leucine. Samples were withdrawn at different times, and the enzyme-forming capacities found that, during conditions which elicit the stringent response in uninfected bacteria, immediate early mRNA is not stringently regulated. This conclusion contradicts the earlier conclusion of others, obtained by measuring incorporation of radioactive uracil; this is explained by the observation of Edlin and Neuhard (1967), confirmed and extended by us to the T4-infected cell, that the incorporation of uracil into RNA of a stringent strain is virtually blocked by amino acid starvation, whereas that of adenine continues at 30 to 50% of the rate seen in the presence of the required amino acid.


Minimal formulation of joint motion for biomechanisms.
Biomechanical systems share many properties with mechanically engineered systems, and researchers have successfully employed mechanical engineering simulation software to investigate the mechanical behavior of diverse biological mechanisms, ranging from biomolecules to human joints. Unlike their man-made counterparts, however, biomechanisms rarely exhibit the simple, uncoupled, pure-axial motion that is engineered into mechanical joints such as sliders, pins, and ball-and-socket joints. Current mechanical modeling software based on internal-coordinate multibody dynamics can formulate engineered joints directly in minimal coordinates, but requires additional coordinates restricted by constraints to model more complex motions. This approach can be inefficient, inaccurate, and difficult for biomechanists to customize. Since complex motion is the rule rather than the exception in biomechanisms, the benefits of minimal coordinate modeling are not fully realized in biomedical research. Here we introduce a practical implementation for empirically-defined internal-coordinate joints, which we call "mobilizers." A mobilizer encapsulates the observations, measurement frame, and modeling requirements into a hinge specification of the permissible-motion manifold for a minimal set of internal coordinates. Mobilizers support nonlinear mappings that are mathematically equivalent to constraint manifolds but have the advantages of fewer coordinates, no constraints, and exact representation of the biomechanical motion-space-the benefits long enjoyed for internal-coordinate models of mechanical joints. Hinge matrices within the mobilizer are easily specified by user-supplied functions, and provide a direct means of mapping permissible motion derived from empirical data. We present computational results showing substantial performance and accuracy gains for mobilizers versus equivalent joints implemented with constraints. Examples of mobilizers for joints from human biomechanics and molecular dynamics are given. All methods and examples were implemented in Simbody™-an open source multibody-dynamics solver available at https://Simtk.org.


Zopiclone: a non-benzodiazepine hypnotic. Controlled comparison to temazepam in insomnia.
Zopiclone, a non-benzodiazepine hypnotic, was compared with temazepam in the treatment of 36 patients suffering from insomnia; the trial period was two weeks, with cross-over of medications at one week and a randomised order of administration. There were highly significant improvements with both drugs, as compared to the control period in respect of: sleep latency, number of times waking during the night, and quality and duration of sleep. The incidence of side-effects was very low. No significant between-drug differences were found in respect to any of the assessment measures or of the incidence of side-effects.


Visible absorption spectrum of the CH3CO radical.
The visible absorption spectrum of the acetyl radical, CH(3)CO, was measured between 490 and 660 nm at 298 K using cavity ring-down spectroscopy. Gas-phase CH(3)CO radicals were produced using several methods including: (1) 248 nm pulsed laser photolysis of acetone (CH(3)C(O)CH(3)), methyl ethyl ketone (MEK, CH(3)C(O)CH(2)CH(3)), and biacetyl (CH(3)C(O)C(O)CH(3)), (2) Cl + CH(3)C(O)H --> CH(3)C(O) + HCl with Cl atoms produced via pulsed laser photolysis or in a discharge flow tube, and (3) OH + CH(3)C(O)H --> CH(3)CO + H(2)O with two different pulsed laser photolysis sources of OH radicals. The CH(3)CO absorption spectrum was assigned on the basis of the consistency of the spectra obtained from the different CH(3)CO sources and agreement of the measured rate coefficients for the reaction of the absorbing species with O(2) and O(3) with literature values for the CH(3)CO + O(2) + M and CH(3)CO + O(3) reactions. The CH(3)CO absorption spectrum between 490 and 660 nm has a broad peak centered near 535 nm and shows no discernible structure. The absorption cross section of CH(3)CO at 532 nm was measured to be (1.1 +/- 0.2) x 10(-19) cm(2) molecule(-1) (base e).


SOX2 and PI3K Cooperate to Induce and Stabilize a Squamous-Committed Stem Cell Injury State during Lung Squamous Cell Carcinoma Pathogenesis.
Although cancers are considered stem cell diseases, mechanisms involving stem cell alterations are poorly understood. Squamous cell carcinoma (SQCC) is the second most common lung cancer, and its pathogenesis appears to hinge on changes in the stem cell behavior of basal cells in the bronchial airways. Basal cells are normally quiescent and differentiate into mucociliary epithelia. Smoking triggers a hyperproliferative response resulting in progressive premalignant epithelial changes ranging from squamous metaplasia to dysplasia. These changes can regress naturally, even with chronic smoking. However, for unknown reasons, dysplasias have higher progression rates than earlier stages. We used primary human tracheobronchial basal cells to investigate how copy number gains in SOX2 and PIK3CA at 3q26-28, which co-occur in dysplasia and are observed in 94% of SQCCs, may promote progression. We find that SOX2 cooperates with PI3K signaling, which is activated by smoking, to initiate the squamous injury response in basal cells. This response involves SOX9 repression, and, accordingly, SOX2 and PI3K signaling levels are high during dysplasia, while SOX9 is not expressed. By contrast, during regeneration of mucociliary epithelia, PI3K signaling is low and basal cells transiently enter a SOX2LoSOX9Hi state, with SOX9 promoting proliferation and preventing squamous differentiation. Transient reduction in SOX2 is necessary for ciliogenesis, although SOX2 expression later rises and drives mucinous differentiation, as SOX9 levels decline. Frequent coamplification of SOX2 and PIK3CA in dysplasia may, thus, promote progression by locking basal cells in a SOX2HiSOX9Lo state with active PI3K signaling, which sustains the squamous injury response while precluding normal mucociliary differentiation. Surprisingly, we find that, although later in invasive carcinoma SOX9 is generally expressed at low levels, its expression is higher in a subset of SQCCs with less squamous identity and worse clinical outcome. We propose that early pathogenesis of most SQCCs involves stabilization of the squamous injury state in stem cells through copy number gains at 3q, with the pro-proliferative activity of SOX9 possibly being exploited in a subset of SQCCs in later stages.


One-carbon metabolism gene polymorphisms and risk of non-Hodgkin lymphoma in Australia.
Dysregulation of the one-carbon metabolic pathway, which controls nucleotide synthesis and DNA methylation, may promote lymphomagenesis. We evaluated the association between polymorphisms in one-carbon metabolism genes and risk of non-Hodgkin lymphoma (NHL) in a population-based case-control study in Australia. Cases (n = 561) and controls (n = 506) were genotyped for 14 selected single-nucleotide polymorphisms in 10 genes (CBS, FPGS, FTHFD, MTHFR, MTHFS, MTR, SHMT1, SLC19A1, TCN1, and TYMS). We also conducted a meta-analysis of all studies of Caucasian populations investigating the association between MTHFR Ex5+79C > T (a.k.a., 677C>T) and NHL risk. A global test of 13 genotypes was statistically significant for diffuse large B-cell lymphoma (DLBCL; P = 0.008), but not for follicular lymphoma (FL; P = 0.27) or all NHL (P = 0.17). The T allele at MTHFR Ex5+79 was marginally significantly associated with all NHL (OR = 1.25, 95% CI = 0.98-1.59) and DLBCL (1.36, 0.96-1.93). The T allele at TYMS Ex8+157 was associated with a reduced risk of FL (0.64, 0.46-0.91). An elevated risk of NHL was also observed among carriers of the G allele at FTHFD Ex21+31 (all NHL, 1.31, 1.02-1.69; DLBCL, 1.50, 1.05-2.14). A meta-analysis of 11 studies conducted in Caucasian populations of European origin (4,121 cases and 5,358 controls) supported an association between the MTHFR Ex5+79 T allele and increased NHL risk (additive model, P = 0.01). In conclusion, the results of this study suggest that genetic polymorphisms of one-carbon metabolism genes such as MTHFR and TYMS may influence susceptibility to NHL.