Height distributions of U.S. children: associations with race, poverty status and parental size.
The stature of children, relative to that of a reference population, is often used as a measure of the nutritional status of a population. But while undernourished children are often small in stature, all small-statured children cannot be assumed to be undernourished as a wide range of hereditary, socioeconomic and health factors also influence growth processes. The distribution of heights of 3850 "healthy" U.S. children 1-11 years of age who participated in the first National Health and Nutrition Examination Survey, a probability sample of the U.S. population, have been examined in relation to age- and sex-specific reference medians, and in relation to race, family income, and height of their parents. The distribution of heights of black children was shifted to the right (i.e., taller) of that of white children (Index of Dissimilarity = 7.9%). The distribution of heights of children of above poverty level income families was shifted to the right of that of children of below poverty level income families (Index of Dissimilarity = 8.4%). In both races, and with family incomes both above and below the poverty income level, the distribution of heights of children of tall parents was shifted to the right of that of children of short parents (Index of Dissimilarity = 20.25%). We concluded that parental stature, economic conditions and race must be considered in interpreting the growth of children in all societies, and before concluding that nutritional factors are the major determinants of short stature.


Broadband coherent anti-Stokes Raman scattering in silicon.
In silicon devices, the spectral bandwidth of coherent anti-Stokes Raman scattering (CARS) is limited by the narrowband nature of the Raman process in this medium. In this Letter, we report the observation of broadband wavelength conversion in silicon through CARS enhanced by the self-phase-modulation-induced spectral broadening of the optical pump. The CARS conversion over a bandwidth substantially greater than the intrinsic Raman linewidth is demonstrated yielding conversion efficiencies as high as 1%. Numerical simulations are performed to explain the spectral features observed in the spectrum of the anti-Stokes signal.


Study on the Mechanism of Fentanyl in Pain Treatment Based on Network Pharmacology.
This is a study that is based on network pharmacology, focusing on the pharmacological mechanism of fentanyl in easing pain. Through PPI, GO, and KEGG network pharmacology, the potential pharmacological mechanism of fentanyl was studied. This study compared and analyzed the overlap between the target genes of the active ingredient of fentanyl and the pain treatment target genes. After constructing PPI based on fentanyl, the GO and KEDD pathways were analyzed for enrichment. On the basis of overlapping genes, we constructed the PPI, GO, and KEGG, and analysis showed that the mechanism was likely to be related to some biological process. This study preliminarily identified the important proteins and metabolic pathways related to fentanyl in pain treatment and expected to provide new evidence and research ideas for the use of fentanyl, enhancing effects and alleviating adverse drug reactions.


Respiratory Health Impacts of Outdoor Air Pollution and the Efficacy of Local Risk Communication in Quito, Ecuador.
Relatively few studies on the adverse health impacts of outdoor air pollution have been conducted in Latin American cities, whose pollutant mixtures and baseline health risks are distinct from North America, Europe, and Asia. This study evaluates respiratory morbidity risk associated with ambient air pollution in Quito, Ecuador, and specifically evaluates if the local air quality index accurately reflects population-level health risks. Poisson generalized linear models using air pollution, meteorological, and hospital admission data from 2014 to 2015 were run to quantify the associations of air pollutants and index values with respiratory outcomes in single- and multi-pollutant models. Significant associations were observed for increased respiratory hospital admissions and ambient concentrations of fine particulate matter (PM2.5), ozone (O3), nitrogen dioxide (NO2), and sulfur dioxide (SO2), although some of these associations were attenuated in two-pollutant models. Significant associations were also observed for index values, but these values were driven almost entirely by daily O3 concentrations. Modifications to index formulation to more fully incorporate the health risks of multiple pollutants, particularly for NO2, have the potential to greatly improve risk communication in Quito. This work also increases the equity of the existing global epidemiological literature by adding new air pollution health risk values from a highly understudied region of the world.


Xanthine oxidase-derived oxygen radicals play significant roles in the development of chronic pancreatitis in WBN/Kob rats.
Although oxygen-derived free radicals are known to play a role in cell injury and DNA alterations, the role of active oxidants in chronic pancreatitis has not been fully elucidated. Using WBN/Kob rats, which spontaneously develop chronic pancreatitis-like lesions, we investigated whether xanthine oxidase (XOD)-derived oxygen radicals are involved in pancreatic tissue injury. WBN/Kob rats were fed a control or a tungsten diet. The latter depletes XOD activity. Histologic al changes, glutathione (GSH) content and XOD and superoxide dismutase (SOD) activities were determined in pancreatic tissue. Pancreatic 8-hydroxy-deoxyguanosine (8-OH-dG) levels and lithostathine mRNA were also examined. In WBN/Kob rats, parenchymal destruction and fibrosis developed at approximately 12 weeks of age and progressed with each month. The activity of XOD was significantly higher in the early period (8-12 weeks), whereas the levels of GSH and SOD decreased after 16 weeks. Levels of 8-OH-dG in WBN/Kob rats were significantly elevated at 16 weeks. Lithostathine mRNA levels started to increase at 8 weeks, but were suppressed at 16 weeks. The tungsten diet significantly attenuated the histological changes in WBN/Kob rats. The increase in pancreatic XOD activity and 8-OH-dG content in WBN/Kob rats was significantly inhibited by the tungsten diet and lithostathine mRNA levels remained high at 16 weeks. These results suggest that oxygen radicals generated by XOD play an important role in oxidative DNA damage and the development of chronic pancreatic injury.


Synthetic Peptides to Target Stringent Response-Controlled Virulence in a Pseudomonas aeruginosa Murine Cutaneous Infection Model.
Microorganisms continuously monitor their surroundings and adaptively respond to environmental cues. One way to cope with various stress-related situations is through the activation of the stringent stress response pathway. In Pseudomonas aeruginosa this pathway is controlled and coordinated by the activity of the RelA and SpoT enzymes that metabolize the small nucleotide secondary messenger molecule (p)ppGpp. Intracellular ppGpp concentrations are crucial in mediating adaptive responses and virulence. Targeting this cellular stress response has recently been the focus of an alternative approach to fight antibiotic resistant bacteria. Here, we examined the role of the stringent response in the virulence of P. aeruginosa PAO1 and the Liverpool epidemic strain LESB58. A ΔrelA/ΔspoT double mutant showed decreased cytotoxicity toward human epithelial cells, exhibited reduced hemolytic activity, and caused down-regulation of the expression of the alkaline protease aprA gene in stringent response mutants grown on blood agar plates. Promoter fusions of relA or spoT to a bioluminescence reporter gene revealed that both genes were expressed during the formation of cutaneous abscesses in mice. Intriguingly, virulence was attenuated in vivo by the ΔrelA/ΔspoT double mutant, but not the relA mutant nor the ΔrelA/ΔspoT complemented with either gene. Treatment of a cutaneous P. aeruginosa PAO1 infection with anti-biofilm peptides increased animal welfare, decreased dermonecrotic lesion sizes, and reduced bacterial numbers recovered from abscesses, resembling the phenotype of the ΔrelA/ΔspoT infection. It was previously demonstrated by our lab that ppGpp could be targeted by synthetic peptides; here we demonstrated that spoT promoter activity was suppressed during cutaneous abscess formation by treatment with peptides DJK-5 and 1018, and that a peptide-treated relA complemented stringent response double mutant strain exhibited reduced peptide susceptibility. Overall these data strongly indicated that synthetic peptides target the P. aeruginosa stringent response in vivo and thus offer a promising novel therapeutic approach.


Semiconducting organic-inorganic nanocomposites by intimately tethering conjugated polymers to inorganic tetrapods.
Semiconducting organic-inorganic nanocomposites were judiciously crafted by placing conjugated polymers in intimate contact with inorganic tetrapods via click reaction. CdSe tetrapods were first synthesized by inducing elongated arms from CdSe zincblende seeds through seed-mediated growth. The subsequent effective inorganic ligand treatment, followed by reacting with short bifunctional ligands, yielded azide-functionalized CdSe tetrapods (i.e., CdSe-N3). Finally, the ethynyl-terminated conjugated polymer poly(3-hexylthiophene) (i.e., P3HT-[triple bond, length as m-dash]) was tethered to CdSe-N3 tetrapods via a catalyst-free alkyne-azide cycloaddition, forming intimate semiconducting P3HT-CdSe tetrapod nanocomposites. Intriguingly, the intimate contact between P3HT and CdSe tetrapod was found to not only render the effective dispersion of CdSe tetrapods in the P3HT matrix, but also facilitate the efficient electronic interaction between these two semiconducting constituents. The successful anchoring of P3HT chains onto CdSe tetrapods was substantiated through Fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopy measurements. Moreover, the absorption and photoluminescence studies further corroborated the intimate tethering between P3HT and CdSe tetrapods. The effect of the type of bifunctional ligands (i.e., aryl vs. aliphatic ligands) and the size of tetrapods on the device performance of hybrid organic-inorganic solar cells was also scrutinized. Interestingly, P3HT-CdSe tetrapod nanocomposites produced via the use of an aryl bifunctional ligand (i.e., 4-azidobenzoic acid) exhibited an improved photovoltaic performance compared to that synthesized with their aliphatic ligand counterpart (i.e., 5-bromovaleric acid). Clearly, the optimal size of CdSe tetrapods ensuring the effective charge transport in conjunction with the good dispersion of CdSe tetrapods rendered an improved device performance. We envision that the click-reaction strategy enabled by capitalizing on two consecutive effective ligand exchanges (i.e., inorganic ligand treatment and subsequent bifunctional ligand exchange) to yield intimately connected organic-inorganic nanocomposites provides a unique platform for developing functional optoelectronic devices.


Bimanual coordination and musical experience: the role of intrinsic dynamics and behavioral information.
Rhythmic interlimb coordination arises from the interaction of intrinsic dynamics and behavioral information, that is, intention, memory, or external information specifying the required coordination pattern. This study investigates the influence of the content of memorized behavioral information on coordination in musically experienced and inexperienced participants. These groups are hypothesized to have different intrinsic dynamics for this task. Stability was assessed in a switching task (variability and switching time). The in-phase, anti-phase, and 90 degrees -phase difference were specified in a neutral and an ecologically relevant manner. Musicians showed more stable coordination than nonmusicians did. No interaction effect was found with memorized behavioral information. Behavioral information showed an interaction effect with phase pattern on coordination variability, with the strongest effect for the 90 degrees -phase pattern. Switching time was affected largely in line with the findings for coordination variability. Participants showed an intraindividual preference for one type of gallop and one type of switch strategy, suggesting different hand roles for the two hands.


Shear-Induced Heteroaggregation of Oppositely Charged Colloidal Particles.
This paper investigates numerically the shear-induced aggregation of mixed populations of colloidal particles leading to the formation of clusters. Suspensions with different amounts of positively and negatively charged colloidal particles are simulated. To resolve the aggregation kinetics and structural properties of the formed clusters, we resort to a mixed deterministic-stochastic simulation method. The method is built on a combination of a Monte Carlo algorithm to sample a statistically expected sequence of encounter events between the suspended particles and a discrete element method built in the framework of Stokesian dynamics to simulate the encounters in a fully predictive manner. Results reveal a strong influence of the composition of the population on both the aggregation kinetics and the aggregate structure. In particular, we observe a size-stabilization phenomenon taking place in the suspension when the relative concentration of the majority particles lies in the range 80-85%; i.e., starting from primary particles, after a short growth period, we observed a cessation of aggregation. Inspection of the aggregate morphology shows that the formed clusters are composed of few minority particles placed in the inner region, while the aggregate surface is covered by majority particles, acting to provide a shielding effect against further growth.


Bell's palsy - part of a polyneuropathy?
Some authors have suggested that Bell's palsy should be considered a part of a polyneuritis cranialis and others, reporting Bell's palsy in patients with diabetes mellitus, proposed that it is an overt sign of a subclinical polyneuropathy. To study whether nerves other than the facial nerve were affected, the clinical findings of 112 patients with Bell's palsy were correlated with a quantitative EMG done on an asymptomatic leg muscle. The electromyographic results obtained from these leg muscles showed changes similar to those occurring in peripheral nerve lesions, and the changes correlated well with the degree of facial palsy. The results indicate that the patients with symptomatic facial dysfunction have an acute subclinical polyneuropathy.


Prediction-based variable selection for component-wise gradient boosting.
Model-based component-wise gradient boosting is a popular tool for data-driven variable selection. In order to improve its prediction and selection qualities even further, several modifications of the original algorithm have been developed, that mainly focus on different stopping criteria, leaving the actual variable selection mechanism untouched. We investigate different prediction-based mechanisms for the variable selection step in model-based component-wise gradient boosting. These approaches include Akaikes Information Criterion (AIC) as well as a selection rule relying on the component-wise test error computed via cross-validation. We implemented the AIC and cross-validation routines for Generalized Linear Models and evaluated them regarding their variable selection properties and predictive performance. An extensive simulation study revealed improved selection properties whereas the prediction error could be lowered in a real world application with age-standardized COVID-19 incidence rates.


Strategies for Building Computing Skills To Support Microbiome Analysis: a Five-Year Perspective from the EDAMAME Workshop.
Here, we report our educational approach and learner evaluations of the first 5 years of the Explorations in Data Analysis for Metagenomic Advances in Microbial Ecology (EDAMAME) workshop, held annually at Michigan State University's Kellogg Biological Station from 2014 to 2018. We hope this information will be useful for others who want to organize computing-intensive workshops and will encourage quantitative skill development among microbiologists.IMPORTANCE High-throughput sequencing and related statistical and bioinformatic analyses have become routine in microbiology in the past decade, but there are few formal training opportunities to develop these skills. A weeklong workshop can offer sufficient time for novices to become introduced to best computing practices and common workflows in sequence analysis. We report our experiences in executing such a workshop targeted to professional learners (graduate students, postdoctoral scientists, faculty, and research staff).


Bilateral Acute Proptosis as Initial Manifestation of Acute Myeloid Leukemia.
The myeloid sarcoma is an extramedullary finding of acute myeloid leukemia (AML), and orbital leukemic tumors occur most commonly during the first decade of life. To our knowledge, we report the youngest patient with bilateral proptosis of both eyes as an initial manifestation of AML. This case highlights the need for peripheral blood smear and neuro-image work-up for acute proptosis in infancy. AML should be considered in the differential diagnosis of an orbital mass, even in the absence of typical leukemic symptoms.


Structural gray matter features and behavioral preliterate skills predict future literacy - A machine learning approach.
When children learn to read, their neural system undergoes major changes to become responsive to print. There seem to be nuanced interindividual differences in the neurostructural anatomy of regions that later become integral parts of the reading network. These differences might affect literacy acquisition and, in some cases, might result in developmental disorders like dyslexia. Consequently, the main objective of this longitudinal study was to investigate those interindividual differences in gray matter morphology that might facilitate or hamper future reading acquisition. We used a machine learning approach to examine to what extent gray matter macrostructural features and cognitive-linguistic skills measured before formal literacy teaching could predict literacy 2 years later. Forty-two native German-speaking children underwent T1-weighted magnetic resonance imaging and psychometric testing at the end of kindergarten. They were tested again 2 years later to assess their literacy skills. A leave-one-out cross-validated machine-learning regression approach was applied to identify the best predictors of future literacy based on cognitive-linguistic preliterate behavioral skills and cortical measures in a priori selected areas of the future reading network. With surprisingly high accuracy, future literacy was predicted, predominantly based on gray matter volume in the left occipito-temporal cortex and local gyrification in the left insular, inferior frontal, and supramarginal gyri. Furthermore, phonological awareness significantly predicted future literacy. In sum, the results indicate that the brain morphology of the large-scale reading network at a preliterate age can predict how well children learn to read.


Circadian phase resetting via single and multiple control targets.
Circadian entrainment is necessary for rhythmic physiological functions to be appropriately timed over the 24-hour day. Disruption of circadian rhythms has been associated with sleep and neuro-behavioral impairments as well as cancer. To date, light is widely accepted to be the most powerful circadian synchronizer, motivating its use as a key control input for phase resetting. Through sensitivity analysis, we identify additional control targets whose individual and simultaneous manipulation (via a model predictive control algorithm) out-perform the open-loop light-based phase recovery dynamics by nearly 3-fold. We further demonstrate the robustness of phase resetting by synchronizing short- and long-period mutant phenotypes to the 24-hour environment; the control algorithm is robust in the presence of model mismatch. These studies prove the efficacy and immediate application of model predictive control in experimental studies and medicine. In particular, maintaining proper circadian regulation may significantly decrease the chance of acquiring chronic illness.


TTY2 genes deletions as genetic risk factor of male infertility.
Y chromosome has a number of genes that are expressed in testis and have a role in spermatogenesis. TTY2L12A and TTY2L2A are the members of testis transcript Y2 (TTY2) that are Y linked multi-copy gene families, located on Yp11 and Yq11 loci respectively. The aim of this study was to investigate frequency of TTY2L12A and TTY2L2A deletions in azoospermic patients compared with fertile males. This study was performed on 45 infertile males with idiopathic azoospermia without any AZF micro deletions (group A), 33 infertile males with azoospermia which do not screened for AZF micro deletions (group B) and 65 fertile males (group C), from October 2013 to April 2015 in west of Iran. Polymerase chain reaction (PCR) method was used for detection of TTY2L12A and TTY2L2A gene deletions in studied groups. No deletions were detected in normal fertile males of group C. 1 out of 45 azoospermic males of group A (2.22%) and 3 out of 33 azoospermic males of group B (9.09%) had TTY2L2A deletion (p= 0.409 and p= 0.036 respectively), also 1 out of 45 azoospermic males of group A (2.22%) and 4 out of 33 azoospermic males of group B (12.12%) had TTY2L12A deletion (p= 0.409 and p= 0.011 respectively). None of azoospermic males in Group A and B had deletions in both genes. Our data showed significant correlation between non-obstructive azoospermia and TTY2L12A and TTY2L2A deletions. Thus, it seems that TTY2L12A and TTY2L2A deletions can consider as one of the genetic risk factors for non-obstructive azoospermia.


Are work well-being variables distinct? A bifactor model of fulfilling work.
The strengths-based inclusive theory of work and psychology of working theory propose that fulfilling work is a key outcome of the vocational intervention. Scholars have further argued that fulfilling work is the holistic experience of well-being in the workplace and can be assessed with meaningful work, work engagement, workplace positive emotions, and job satisfaction. This theoretical perspective suggests a bifactor model would best explain the relations among these variables, but this claim remains untested. Therefore, the purpose of this study was to investigate whether a bifactor model best explained the relations among the four components of fulfilling work, in comparison to other plausible models. We also examined the concurrent and convergent validity of the fulfilling work construct, using other well-being variables, symptoms of distress, and contextual factor variables drawn from vocational theories. Supporting hypotheses, we found that a bifactor model best fit the data. We also found that fulfilling work positively related to eudaimonic work well-being, hedonic work well-being, and life satisfaction and negatively related to symptoms of distress. Finally, fulfilling work positively related to income and subjective social class. These findings offer conceptual and statistical implications of fulfilling work for research, counseling, organizations, and social advocacy. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


What criteria do consumer health librarians use to develop library collections? a phenomenological study.
The criteria for determining whether resources are included in consumer health library collections are summarized in institutional collection development policies (CDPs). Evidence suggests that CDPs do not adequately capture all of these criteria. The aim of this study was to describe the resource review experience of librarians and compare it to what is described in CDPs. A phenomenological approach was used to explore and describe the process. Four consumer health librarians independently evaluated cancer-related consumer health resources and described their review process during a semi-structured telephone interview. Afterward, these librarians completed online questionnaires about their approaches to collection development. CDPs from participating libraries, interview transcripts, and questionnaire data were analyzed. Researchers summarized the findings, and participating librarians reviewed results for validation. Librarians all utilized similar criteria, as documented in their CDPs; however, of thirteen criteria described in the study, only four were documented in CDPs. CDPs for consumer health libraries may be missing important criteria that are considered integral parts of the collection development process. A better understanding of the criteria and contextual factors involved in the collection development process can assist with establishing high-quality consumer health library collections.


Fabry disease: renal sonographic and magnetic resonance imaging findings in affected males and carrier females with the classic and cardiac variant phenotypes.
To describe the renal ultrasonography (US) and magnetic resonance imaging (MRI) findings in affected males and female carriers with the classic and cardiac variant phenotypes of Fabry disease (alpha-galactosidase A [alpha-Gal A] deficiency). The renal US and MRI features of 76 classically affected males (aged 7-53 years), 40 female carriers from classically affected families (aged 18-66 years), and 6 males with the cardiac variant phenotype (aged 17-59 years) were reviewed by 3 blinded board-certified radiologists. The images were evaluated for the presence of cortical cysts, parapelvic cysts, renal atrophy, decreased cortical thickness, increased echogenicity (US only), and decreased corticomedullary differentiation (MRI only). The consensus findings were analyzed with respect to the patients' sex, age, Fabry genotype and phenotype, and renal function. MRI was more sensitive than US in detecting radiographic abnormalities. In the 76 classically affected males, the most common US abnormalities were cysts (36.9%; cortical cysts = 22.4%, parapelvic cysts = 14.5%), increased echogenicity (17.1%), and decreased cortical thickness (11.9%), whereas the most common MRI abnormalities were cysts (47.3%; cortical cysts = 28.9%, parapelvic cysts = 18.4%), loss of corticomedullary differentiation (43.4%), and decreased cortical thickness (7.9%). Among the 40 female carriers, common US abnormalities included cysts (20%; cortical cysts = 10%, parapelvic cysts = 10%) and increased echogenicity (7.5%), whereas MRI findings included decreased corticomedullary differentiation (40%) and cysts (37.5%; cortical cysts = 20%; parapelvic cysts = 17.5%). Renal US and MRI were normal in 5 classically affected males aged 12 years or younger and 2 female carriers aged 20 years or younger. Among the 6 male cardiac variants, abnormal US findings included cysts (66.3%; cortical cysts = 50%, parapelvic cysts = 16.3%) and increased echogenicity (33.3%), whereas MRI detected decreased corticomedullary differentiation in all (100%) and cysts in 83% (cortical cysts = 66.7%; parapelvic cysts = 16.3%). Serum creatinine levels were elevated (>1.2 mg/dL) in 40.8% and 15% of the classically affected males and female carriers with US and/or MRI abnormalities compared with 14.8% and 0%, respectively, who had elevated serum creatinine levels but no detectable radiographic abnormalities. There was no association of alpha-Gal A genotype with type or frequency of abnormalities in classically affected patients. Among classically affected males and female carriers, renal US and/or MRI abnormalities were detected in 64.5% and 60%, respectively. The occurrence and number of abnormalities increased with age in affected males and female carriers. Cysts, particularly parapelvic cysts, were more common and appeared earlier than in the general population. No renal abnormalities were detected in classically affected males or female carriers <12 years or <20 years of age, respectively. Five of the 6 males with the later-onset milder cardiac variant phenotype had loss of corticomedullary differentiation on MRI. Renal imaging abnormalities were more frequent in older patients with elevated serum creatinine levels, regardless of alpha-Gal A genotype or Fabry phenotype.


Intraoperative Neurophysiological Monitoring (Motor and Somatosensory Evoked Potentials) in Dorsal Root Entry Zone Lesioning for Brachial Plexus Avulsion Pain.
To address the feasibility and importance of intraoperative neurophysiological monitoring (IONM) in dorsal root entry zone (DREZ) lesioning for brachial plexus avulsion pain. Muscle motor evoked potential (mMEP) and somatosensory evoked potential (SSEP) were applied during DREZ lesioning for brachial plexus avulsion pain. IONM of mMEPs and SSEPs was feasible for monitoring of the spinal cord during DREZ lesioning. With the exception of 3 unrecordable mMEPs in ipsilateral arms, mMEPs were preserved and referenced to look for changes according to lesioning in 6 upper extremities (66.6%) and 8 lower extremities. All 3 patients with >50% reduction in baseline mMEP amplitude after lesioning in either the ipsilateral upper or lower extremities showed postoperative ataxia and weakness of the lower extremities (100%). Only 2 out of 9 patients (22.2%) with brachial plexus avulsion pain had discernible baseline SSEPs in the ipsilateral upper extremities. One of 2 patients with discernible SSEPs in the upper extremities showed significant SSEP events during the DREZ lesioning and experienced postoperative ataxia and weakness in the legs despite the absence of a SSEP event in the lower extremities. Significant events on IONM were common during DREZ lesioning for brachial plexus avulsion pain and were closely related to the occurrence of postoperative neurological deficits.


Idiopathic intracranial hypertension associated with iron deficiency anaemia: a lesson for management.
To document the causal association of iron deficiency anaemia (IDA) and intracranial hypertension (IH). A consecutive case note review of patients with a clinical diagnosis of idiopathic intracranial hypertension (IIH) and anaemia presenting to a tertiary referral unit over a 2.5-year period. Demographics, aetiology and clinical details were recorded and analysed. Eight cases were identified from 77 new cases presenting with IIH. All 8 had documented microcytic anaemia with clinical evidence of raised intracranial pressure. There was no evidence of venous sinus thrombosis on MRI and MR venography in 7 subjects and on repeated CT venography in 1. On correction of anaemia alone, 7 cases resolved. One patient with severe progressive visual loss underwent ventriculoperitoneal shunt in addition to treatment of anaemia, with good outcome. The incidence of this association is 10.3%. These cases present an association between IDA and IH, in the absence of cerebral sinus thrombosis. As a clinically significant proportion of cases presenting with signs of IIH have IDA, we recommend all patients presenting with IIH have full blood counts and if they are found to be anaemic, they should be treated appropriately.


Application of Molecular Methods for Carbapenemase Detection.
This study has evaluated the correlation between different carbapenemases detection methods on carbapenem non-susceptible Klebsiella pneumoniae strains from Northern and Eastern Europe; 31 institutions in 9 countries participated in the research project, namely Finland, Estonia, Latvia, Lithuania, Russia, St. Petersburg, Poland, Belarus, Ukraine, and Georgia. During the research program, a total of 5,001 clinical K. pneumoniae isolates were screened for any carbapenem non-susceptibility by the disk diffusion method, Vitek 2 or Phoenix system following the EUCAST guideline on detection of resistance mechanisms, version 1.0. Strains isolated from outpatients and hospitalized patients from April 2015 to June 2015 were included. All types of samples (blood, pus, urine, etc.) excluding fecal screening or fecal colonization samples have been represented. In total, 171 carbapenemase screening-positive K. pneumoniae isolates (3.42%) were found and characterized. Several methods were used for detection of carbapenemases production, including Luminex assay (PCR and hybridization), whole genome sequencing, MALDI-TOF based Imipenem degradation assay, and immunochromatography testing. Minimal inhibitory concentration determination for Meropenem by agar-based gradient method was also used. Finally, 83 K. pneumoniae strains were carbapenemase negative by all confirmation methods (49.4% of all screening-positive ones), 74 - positive by three methods (44.0%), 8 - positive by two methods (4.8%) and 3 - positive by only one method (1.8%). The sensitivity of the tests was 96.3% for Whole genome sequencing and MALDI-TOF assay (both three undetected cases), and 95.1% for Luminex-Carba (4 undetected cases). The most commonly detected carbapenemases were NDM (n = 54) and OXA-48 (n = 26), followed by KPC-2, VIM-5, and OXA-72 (one case of each). Our results showed that different types of carbapenemases can be detected in the countries involved in the project. The sensitivity of our methods for carbapenemase detection (including screening as a first step and further confirmation tests) was >95%, but we would recommend using different methods to increase the sensitivity of detection and make it more precise.


Tanshinone IIA Activates Nuclear Factor-Erythroid 2-Related Factor 2 to Restrain Pulmonary Fibrosis via Regulation of Redox Homeostasis and Glutaminolysis.
Pulmonary fibrosis (PF) is characterized by myofibroblast activation through oxidative stress. However, the precise regulation of myofibroblast transdifferentiation remains largely uncharacterized. In this study, we found that tanshinone IIA (Tan-IIA), an active component in the root of Salvia miltiorrhiza Bunge, can suppress reactive oxygen species (ROS)-mediated activation of myofibroblast and reduce extracellular matrix deposition in bleomycin (BLM)-challenged mice through the regulation of nuclear factor-erythroid 2-related factor 2 (Nrf2). Additionally, Tan-IIA restored redox homeostasis by upregulating Nrf2 with NADPH oxidase 4 suppression and effectively prevented myofibroblast activation by blocking ROS-mediated protein kinase C delta (PKCδ)/Smad3 signaling. Nrf2 knockdown in the fibroblasts and the lungs of BLM-treated mice reduced the inhibitory effects of Tan-IIA, indicating the essential role of Nrf2 in the Tan-IIA activity. Tan-IIA impaired the binding of kelch-like ECH-associated protein 1 (Keap1) to Nrf2 by promoting the degradation of Keap1 and thereby increasing Nrf2 induction by protecting Nrf2 stability against ubiquitination and proteasomal degradation. Importantly, we also found that the glutamate anaplerotic pathway was involved in energy generation and biosynthesis in activated myofibroblasts and their proliferation. Tan-IIA shunted glutaminolysis into glutathione (GSH) production by activating Nrf2, resulting in the reduction of glutamate availability for tricarboxylic acid cycle. Ultimately, myofibroblast activation was prevented by impairing cell proliferation. Innovation and Conclusion: In addition to the regulation of redox homeostasis, our work showed that Tan-IIA activated Nrf2/GSH signaling pathway to limit glutaminolysis in myofibroblast proliferation, which provided further insight into the critical function of Nrf2 in PF.


Technical note: evaluation of data loggers for measuring lying behavior in dairy calves.
Lying behavior might indicate how the animal interacts with its environment and is an important indicator of cow and calf comfort. Measuring behavior can be time consuming; therefore, behavioral recording with the help of loggers has become common. Recently, the Hobo Pendant G data logger (Onset Computer Corp., Bourne, MA) was validated for measuring lying behavior in cows but no work to date has validated this logger for measuring lying behavior in calves. The objective of this study was to test the accuracy of the Hobo Pendant G data logger for measuring total lying time and frequency of lying bouts in dairy calves. In 2 experiments (experiment 1: thirty-seven 2-h observation periods; experiment 2: nineteen 24-h observation periods), we tested the effect of 2 different recording intervals, the effect of attachment to different legs, and the effect of removing short, potentially erroneous readings. We found an excellent relationship when comparing the 30-s and 60-s recording intervals. For total lying time and bout frequency, the highest correlation was found when the logger was attached to the hind legs and recording was conducted with a 60-s sampling interval. In experiment 2, average total lying time was 1,077 ± 54 min/24 h (18.0 ± 0.9h/24h), with an average frequency of 19.4 ± 4.5 bouts per day. Predictability, sensitivity, and specificity for experiment 2 were >97% using the 60-s recording interval and removing single readings of lying or standing from the data set compared with direct observation as reference. The data logger accurately measured total lying time and bout frequency when the sampling interval was ≤ 60 s and short readings of lying and standing up to 1 min were converted into the preceding behavior. The best results were achieved by attaching the logger to the right hind leg.


Versican, matrix Gla protein, and death-associated protein expression affect muscle satellite cell proliferation and differentiation.
Our previous transcriptional profiling study using a turkey skeletal muscle-specific oligonucleotide microarray revealed over 3,000 genes that were differentially expressed at 3 critical stages of muscle development: 18 d embryonic, 1 d posthatch, and 16 wk of age. The genes versican, matrix Gla protein (MGP), and death-associated protein (DAP) were selected to study for their potential effects on muscle satellite cell proliferation and differentiation, as their functions in other tissues are suggestive of possible key roles in the regulation of myogenesis and they are differentially expressed throughout muscle development in the turkey. Using small interfering RNA to knockdown the expression of these genes during proliferation and differentiation, each of the genes was found to differentially affect proliferation and differentiation. Versican and MGP predominantly affected proliferation with line effects, but later stages of differentiation were affected by the knockdown of versican and MGP. The underexpression of DAP inhibited myotube formation, which is a necessary stage in the development of muscle fibers. Without myotube development, muscle fiber formation will be inhibited or abolished. This is the first report that these genes with no previously documented functions with regard to muscle development play a critical role in muscle cell proliferation and differentiation.


Migratory constraints on yolk precursors limit yolk androgen deposition and underlie a brood reduction strategy in rockhopper penguins.
Hormonally mediated maternal effects link maternal phenotype and environmental conditions to offspring phenotype. The production of lipid-rich maternal yolk precursors may provide a mechanism by which lipophilic steroid hormones can be transported to developing yolks, thus predicting a positive correlation between yolk precursors in mothers and androgen levels in eggs. Using rockhopper penguins (Eudyptes chrysocome), which produce a two-egg clutch characterized by extreme egg-size dimorphism, reversed hatching asynchrony and brood-reduction, we examined correlations between circulating concentrations of the primary yolk-precursor vitellogenin (VTG) and levels of yolk androgens. Previous work in Eudyptes penguins has shown that egg-size dimorphism is the product of migratory constraints on yolk precursor production. We predicted that if yolk precursors are constrained, androgen transport to developing yolks would be similarly constrained. We reveal positive linear relationships between maternal VTG and androgens in small A-eggs but not larger B-eggs, which is consistent with a migratory constraint operating on the A-egg. Results suggest that intra-clutch variation in total yolk androgen levels depends on the production and uptake of yolk precursors. The brood reduction strategy common to Eudyptes might thus be best described as the result of a migratory constraint.


Microbial communities present in the lower respiratory tract of clinically healthy birds in Pakistan.
Commercial poultry is an important agricultural industry worldwide. Although dense living conditions and large flocks increase meat and egg production, they also increase the risk of disease outbreaks and zoonoses. Current pathogen identification methods mostly rely on culture-dependent techniques and, therefore, are limited to a very small number of bacteria present in the environment. Next Generation Sequencing allows for culture-independent characterization of lower respiratory microbiome of birds including the identification of novel commensals and potentially emerging pathogens. In this study, we collected tracheo-bronchoalveolar lavage of 14 birds raised at 3 different farms in the Punjab province of Pakistan. To characterize the lower respiratory microbiome of these birds, we sequenced hyper-variable regions of the 16S ribosomal subunit gene. Although dominated by bacteria belonging to a small number of taxonomic classifications, the lower respiratory microbiome from each farm was far more diverse and novel than previously known. The differences in microbiome among farms suggest that inter-farm differences affect the microbiome of birds more than breed, geographic location, or management system. The presence of potential and known pathogens in genetically similar specialty breeds of chickens kept at unnaturally high densities and under variable conditions presents an extraordinary opportunity for the selection of highly pathogenic bacteria. In some instances, opportunistic respiratory pathogens were observed in apparently healthy birds. Understanding and monitoring the respiratory microbiome of such populations may allow the early detection of future disease threats.


Melanocytic Neoplasms With MAP2K1 in Frame Deletions and Spitz Morphology.
With the advent of better molecular characterization of Spitz melanocytic neoplasms, there has been increasing effort to better understand and describe the relationships between specific driver fusion and/or mutations with the clinical and histomorphological characteristics of the lesions. Structural rearrangements in mitogen activated protein kinase genes have recently been noted to be important in Spitz neoplasms. Only very few reports, however, have described in detail melanocytic tumors with in frame deletions in MAP2K1. Cases in the literature with this aberration have been described as having a diagnosis of Spitz, deep penetrating nevi, or pigmented epithelioid melanocytoma. In this study, we describe a cohort of 6 cases with MAP2K1 activating in frame deletions. The morphologic spectrum of the cases was broad. Common features of these cases include Spitzoid cytomorphology (5/6) cases, prominent melanin pigmentation (4/6) cases, and deep penetrating nevi-like plexiform architecture (3/6) cases. The diagnoses at the time of clinical care of these cases included nevus of Reed (1/6), desmoplastic Spitz tumor (1/6), BAPoma (1/6), deep penetrating melanocytic nevus (2/6), and melanoma (1/6). Clinical follow-up was available in 3 of the 6 cases. None of the patients had a tumor recurrence. This builds on the growing literature to help expand the spectrum of changes associated with Spitzoid melanocytic neoplasms.


Subacute osteomyelitis of the femur due to Fusobacterium nucleatum in a 7-year-old boy.
Subacute hematogenous osteomyelitis is an insidious infection, which commonly has a delayed diagnosis. We describe the case of a 7-year-old boy with subacute osteomyelitis, which was initially considered to be a bone tumor. Infection should be considered in all cases of bone pain, especially in children, even in the absence of typical systemic features of inflammation.


Methanogenic diversity and activity in hypersaline sediments of the centre of the Napoli mud volcano, Eastern Mediterranean Sea.
Submarine mud volcanoes are a significant source of methane to the atmosphere. The Napoli mud volcano, situated in the brine-impacted Olimpi Area of the Eastern Mediterranean Sea, emits mainly biogenic methane particularly at the centre of the mud volcano. Temperature gradients support the suggestion that Napoli is a cold mud volcano with moderate fluid flow rates. Biogeochemical and molecular genetic analyses were carried out to assess the methanogenic activity rates, pathways and diversity in the hypersaline sediments of the centre of the Napoli mud volcano. Methylotrophic methanogenesis was the only significant methanogenic pathway in the shallow sediments (0-40 cm) but was also measured throughout the sediment core, confirming that methylotrophic methanogens could be well adapted to hypersaline environments. Hydrogenotrophic methanogenesis was the dominant pathway below 50 cm; however, low rates of acetoclastic methanogenesis were also present, even in sediment layers with the highest salinity, showing that these methanogens can thrive in this extreme environment. PCR-DGGE and methyl coenzyme M reductase gene libraries detected sequences affiliated with anaerobic methanotrophs (mainly ANME-1) as well as Methanococcoides methanogens. Results show that the hypersaline conditions in the centre of the Napoli mud volcano influence active biogenic methane fluxes and methanogenic/methylotrophic diversity.


Implementation of electro-optic spectral shearing interferometry for ultrashort pulse characterization.
Electro-optic spectral shearing interferometry is implemented for the highly sensitive characterization of ultrashort pulses from a free-running source. A simple phase-locked-loop clock recovery circuit is used to extract the high-frequency microwave clock signal to drive the phase modulator for spectral shearing. We demonstrate accurate full temporal characterization of 200-fs pulses from an optical parametric oscillator at 5-pW average power.


Belliella aquatica sp. nov., isolated from a saline lake.
A Gram-staining-negative bacterium, strain TS-T86(T), was isolated from Lake Tuosu, a saline lake (salinity 5.4%, w/w) in Qaidam basin, China. Its taxonomic position was determined by using a polyphasic approach. Strain TS-T86(T) was strictly heterotrophic, aerobic and catalase- and oxidase-positive. Cells were non-spore-forming, non-motile rods, 0.4-0.6 µm wide and 1.2-2.3 µm long. Growth was observed in the presence of 0-9.0% (w/v) NaCl (optimum, 2.0%), at 4-35 °C (optimum, 25 °C) and at pH 7.0-10.5 (optimum, pH 8.5-9.0). Strain TS-T86(T) contained MK-7 as the predominant respiratory quinone. The major fatty acids (>10%) were iso-C15 : 1 G, iso-C15 : 0, iso-C17 : 1ω9c, iso-C17 : 0 3-OH and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c). The polar lipids consisted of phosphatidylethanolamine, an unknown phospholipid, six unidentified aminolipids and two uncharacterized lipids. The DNA G+C content was 35 mol% (T m). Phylogenetic trees based on 16S rRNA gene sequences showed that strain TS-T86(T) was associated with the genus Belliella, and showed the highest sequence similarity to Belliella baltica BA134(T) (98.5 %) and then to Belliella kenyensis No.164(T) (95.7%) and Belliella pelovolcani CC-SAL-25(T) (95.3 %). DNA-DNA relatedness of strain TS-T86(T) to Belliella baltica DSM 15883(T) was 32 ± 3%. It is concluded that strain TS-T86(T) represents a novel species of the genus Belliella, for which the name Belliella aquatica sp. nov. is proposed. The type strain is TS-T86(T) ( = CGMCC 1.12479(T) = JCM 19468(T)).


CTG expansion & haplotype analysis in DM1 gene in healthy Iranian population.
Myotonic dystrophy type 1 (DM1) is due to an unstable expansion of CTG repeat in the DMPK gene (19q13.3). The CTG repeat is highly polymorphic (5 to 37) in healthy individuals. According to the hypothesis that expanded (CTG)n alleles originated from larger normal alleles, there may exist a correlation between the prevalence of DM1 and the frequency of large size normal alleles. Strong linkage disequilibrium between different length alleles and the three biallelic markers, Alu, Hinf1 and Taq1, has been reported. To determine the distribution of normal alleles, the frequency of larger normal alleles and analysis of the three biallelic markers, in healthy Iranian controls. Polymerase chain reaction (PCR) was conducted on two hundred unrelated healthy individuals from different ethnic groups living in Iran to determine the size of the alleles. Markers were analyzed by PCR/RFLP on 174 chromosomes from other control healthy individuals. Our data reveals that 23.7% of alleles had 5 CTG repeats and 7.2% of alleles had > 18 CTG repeats. The analysis of haplotypes revealed that 75% of CTG5 and 80% of CTG > 18 had the (+++) haplotype. The frequency of alleles with CTG > 18 in Iran is similar to that of Western Europe and Japan.


The effect of inotropic and/or vasopressor support on postoperative survival following equine colic surgery.
To determine the effects of dobutamine and phenylephrine on intra- and postoperative survival in horses undergoing emergency abdominal surgery. Retrospective case analysis. A total of 637 client-owned horses undergoing colic surgery. Clinical details of horses admitted for colic surgery were recorded on a computer database. Information collected included history, clinical variables observed before surgery, anaesthesia and surgical details, and postoperative survival and morbidity rates. Details of specific importance for this study were those recorded during anaesthesia, in particular the duration of dobutamine and phenylephrine administration, separately and combined, and total anaesthesia time. Two outcomes were considered: 1) intra-operative death, i.e. death between time of pre-anaesthetic medication and recovery from anaesthesia (defined as horse walking from recovery box); and 2) all deaths, i.e. death at any time after induction of anaesthesia. The definition of 'death' included euthanasia. Univariable and multivariable statistical analyses were performed to evaluate the associations between dobutamine and/or phenylephrine use and these two outcomes. Results from univariable analyses suggested that dobutamine administration was not significantly associated with increased intra- or postoperative mortality. Phenylephrine administration showed univariable association with intra- and postoperative death. However, in multivariable models adjusted for the effects of heart rate and packed cell volume at admission, the phenylephrine effect was not significantly associated with intra-operative, or other types of death. This study provides no evidence to suggest that dobutamine or phenylephrine administration is associated with altered survival rates during or after colic surgery. Our study supports previous work, suggesting that pre-existing cardiovascular status is an important prognostic determinant in equine colic cases. It provides no evidence that dobutamine or phenylephrine administration is associated with survival.


[Functionally important lysine residues in inorganic pyrophosphatase from E. coli. I. Interaction of inorganic pyrophosphatase with pyridoxal-5'-phosphate].
Interaction of inorganic pyrophosphatase from E. coli with pyridoxal-5'-phosphate includes binding of the reagent at the active site through the phosphate group and then a reversible modification of one lysine residue in each of the enzyme's subunit. In the equilibrium state the protein's molecules contain both inactive modified and native subunits. A stable secondary amine is formed upon the sodium borohydride reduction of the modified protein.


Identification of harmless and pathogenic algae of the genus Prototheca by MALDI-MS.
The only plants infectious for mammals, green algae from the genus Prototheca, are often overseen or mistaken for yeast in clinical diagnosis. To improve this diagnostical gap, a method was developed for fast and reliable identification of Prototheca. A collection of all currently recognized Prototheca species, most represented by several strains, were submitted to a simple extraction by 70% formic acid and ACN; the extracts were analyzed by means of MALDI-MS. Most of the peaks were found in the range from 4 to 20 kDa and showed a high reproducibility, not in absolute intensities, but in their peak pattern. The selection of measured peaks is mostly due to the technique of ionization in MALDI-MS, because proteins in the range up to 200 kDa were detected using gel electrophoresis. Some of the proteins were identified by peptide mass fingerprinting and MS(2) analysis and turned out to be ribosomal proteins or other highly abundant proteins such as ubiquitin. For the preparation of a heatmap, the intensities of the peaks were plotted and a cluster analysis was performed. From the peak-lists, a principal component analysis was conducted and a dendrogram was built. This dendrogram, based on MALDI spectra, was in fairly good agreement with a dendrogram based on sequence information from 18S DNA. As a result, pathogenic and nonpathogenic species from the genus Prototheca can be identified, with possible consequences for clinical diagnostics by MALDI-typing.


Organization and Formation of the Crossed-Foliated Biomineral Microstructure of Limpet Shells.
To construct their shells, molluscs are able to produce a large array of calcified materials including granular, prismatic, lamellar, fibrous, foliated, and plywood-like microstructures. The latter includes an aragonitic (the crossed-lamellar) and a calcitic (the crossed-foliated) variety, whose modes of formation are particularly enigmatic. We studied the crossed-foliated calcitic layers secreted solely by members of the limpet family Patellidae using scanning and transmission electron microscopy and electron backscatter diffraction. From the exterior to the interior, the material becomes progressively organized into commarginal first-order lamellae, with second and third order lamellae dipping in opposite directions in alternating lamellae. At the same time, the crystallographic texture becomes stronger because each set of the first order lamellae develops a particular orientation for the c-axis, while both sets maintain common orientations for one {104} face (parallel to the growth surface) and one a-axis (perpendicular to the planes of the first order lamellae). Each first order lamella shows a progressive migration of its crystallographic axes with growth in order to adapt to the orientation of the set of first order lamellae to which it belongs. To explain the progressive organization of the material, we hypothesize that a secretional zebra pattern, mirrored by the first order lamellae on the shell growth surface, is developed on the shell-secreting mantle surface. Cells belonging to alternating stripes behave differently to determine the growth orientation of the laths composing the first order lamellae. In this way, we provide an explanation as to how plywood-like materials can be fabricated, which is based mainly on the activity of mantle cells.


Arabidopsis SMG7 protein is required for exit from meiosis.
Meiosis consists of two nuclear divisions that are separated by a short interkinesis. Here we show that the SMG7 protein, which plays an evolutionarily conserved role in nonsense-mediated RNA decay (NMD) in animals and yeast, is essential for the progression from anaphase to telophase in the second meiotic division in Arabidopsis. Arabidopsis SMG7 is an essential gene, the disruption of which causes embryonic lethality. Plants carrying a hypomorphic smg7 mutation exhibit an elevated level of transcripts containing premature stop codons. This suggests that the role of SMG7 in NMD is conserved in plants. Furthermore, hypomorphic smg7 alleles render mutant plants sterile by causing an unusual cell-cycle arrest in anaphase II that is characterized by delayed chromosome decondensation and aberrant rearrangement of the meiotic spindle. The smg7 phenotype was mimicked by exposing meiocytes to the proteasome inhibitor MG115. Together, these data indicate that SMG7 counteracts cyclin-dependent kinase (CDK) activity at the end of meiosis, and reveal a novel link between SMG7 and regulation of the meiotic cell cycle.


Involvement of an NAD(P)H oxidase-like enzyme in superoxide anion and hydrogen peroxide generation by rat type II cells.
Although alveolar macrophages are considered to be the primary cellular mediators of host defence in the lung, there is increasing evidence that type II cells may also play an active role in host defence. A study was undertaken to investigate whether type II cells generate O2-. and H2O2 via an NADPH oxidase-like system and whether exposure of the type II cells to soluble or particulate stimuli known to activate NADPH oxidase in macrophages also leads to increased production of H2O2. Rat type II cells and alveolar macrophages were exposed to 10, 100, or 1000 nM phorbol-12-myristate-13-acetate (PMA) and the production of O2-. and H2O2 was determined by chemiluminescence. Thirty minutes before stimulation with 1 microM PMA type II cells were also exposed to the same concentrations of a protein kinase C (PKC) antagonist GF109203x, the non-selective protein kinase inhibitor staurosporine (1, 10, or 100 nM), or the NADPH oxidase inhibitor diphenyliodonium chloride (DPI) (1, 10, 100, or 1000 microM). The effects of arachidonic acid, zymosan and Staphylococcus aureus on H2O2 production were determined. Cell membrane fractions from type II cells and macrophages were assayed for NADPH oxidase activity. After exposure to 1 microM PMA, O2-. and H2O2 generation increased 6.3-fold and 9.0-fold, respectively, in type II cells and 2.4-fold and 5.2-fold, respectively, in macrophages. In contrast to the macrophages, the increase in O2-. and H2O2 generation by type II cells was completely prevented by 1 mM KCN. Preexposure to GF109203x, staurosporine, or DPI completely prevented the rise in O2-. and H2O2 generation. Mean (SD) NADPH oxidase activity of 138 (38) nmol O2-./min/mg protein was found in membrane fraction I of the type II cells, and 102 (31) nmol O2-./min/mg protein in fraction II. Macrophages showed higher NADPH oxidase activity in membrane fraction II. In type II cells exposure to arachidonic acid led to a significant 5.3-fold increase in H2O2 generation, exposure to zymosan increased H2O2 generation 46-fold, and exposure to S aureus 25-fold with a maximum 30-50 minutes after addition of the bacteria. Type II cells generate O2-. and H2O2 via a PKC-mediated activation of an NAD(P)H oxidase-like membrane bound enzyme. Arachidonic acid, zymosan, and bacteria also give rise to increased H2O2 production. Type II cells might thus play an active role in host defence.


Extracellular poly(alpha-L-guluronate)lyase from Corynebacterium sp.: purification, characteristics, and conformational properties.
Extracellular alginate lyase was purified from the culture supernatant of Corynebacterium sp. isolated from the sewage of a sea tangle processing factory in order to elucidate the structure-function relationship of alginate lyase. The electrophoretically homogeneous enzyme was shown to have a molecular mass of 27 kDa by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) and by gel filtration, with an isoelectric point of 7.3. The molecular mass from amino acid analysis was 28.644 kDa. The optimal pH and temperature for the enzyme reaction were around 7.0 and 55 degrees C, respectively. Metal compounds such as MnCl2 and NiCl2 increased the enzyme activity. The enzyme was identified as the endolytic poly(alpha-L-guluronate)lyase, which was active on poly(alpha-L-1,4-guluronate) and caused a rapid decrease in the viscosity of alginate solution. Measurement of the far-UV circular dichroic spectrum of the enzyme molecule gave a spectrum with a deep trough at 215 nm accompanied by a shallow one at around 237 nm, and with a high peak at 197 nm and a much lower one at 230 nm. This spectrum was most likely to be that of the beta-form of the enzyme molecule and resembled poly(beta-D-mannuronate)lyase from Turbo cornutus (wreath shell) and poly(alpha-L-guluronate)lyase from Vibrio sp. (marine bacterium). The near-UV circular dichroic spectrum was characteristic for aromatic amino acid residues. In the presence of 6 M urea, these spectra changed drastically in the near-UV and a little in the far-UV with the disappearance of the enzyme activity. Removal of the denaturant in the enzyme solution by dialysis restored both the activity and inherent circular dichroic spectra. The beta-sheets observed in alginate lyases as the major ordered structure seem to be a common conformation for the lyases.


Working to Eliminate Cancer Health Disparities from Tobacco: A Review of the National Cancer Institute's Community Networks Program.
In 2005, the National Cancer Institute funded the Community Networks Program (CNP), which aimed to reduce cancer health disparities in minority racial/ethnic and underserved groups through community-based participatory research, education, and training. The purpose of this study was to describe the CNP model and their tobacco-related work in community-based research, education, and training using a tobacco disparities research framework. We conducted a comprehensive review of the CNP tobacco-related activities including publications, published abstracts, research activities, trainee pilot studies, policy-related activities, educational outreach, and reports produced from 2005-2009. Two authors categorized the tobacco-related activities and publications within the framework. Although there was no mandate to address tobacco, the CNPs produced 103 tobacco-related peer-reviewed publications, which reflects the largest proportion (12%) of all CNP cancer-related publications. Selected publications and research activities were most numerous under the framework areas "Psychosocial Research," "Surveillance," "Epidemiology," and "Treatment of Nicotine Addiction." Thirteen CNPs participated in tobacco control policymaking in mainstream efforts that affected their local community and populations, and 24 CNPs conducted 1147 tobacco-related educational outreach activities. CNP activities that aimed to build research and infrastructure capacity included nine tobacco-related pilot projects representing 16% of all CNP cancer-related pilot projects, and 17 publications acknowledging leveraged partnerships with other organizations, a strategy encouraged by the CNP. The CNP is a promising academic-community model for working to eliminate tobacco-related health disparities. Future efforts may address scientific gaps, consider collaboration across groups, assess the extent of operationalizing community-based participatory research, and improve common tracking measures.


[Liberation of H3-GABA from isolated nerve endings of the cerebral cortex of rats under the effect of psychotropic drugs].
The influence of a number of neuroleptics and antidepressants on the release of 3H-GABA from the synaptosomas of the cerebral cortex in rats was studied. Chlorpromazine, phthorphenazine, trifluperidol, imipramine, phthoracizine caused an intensified liberation of 3H-GABA from the synaptosomas. In this phenothiazine neuroleptics proved more active than did trifluperidol and antidepressants. New neuroleptics--azabuperon and carbidine and also diphenylhydantoin tended to restrict the liberation of the mediator from synaptosomas. The tranquilizer diazepam, gamma-oxybutyric acid and carbamazepine failed to have any significnat effect on this process. The described effects of thepsychotropic substances are due, apparently, to their direct influence on the synaptomal membrane.


[The role of suprasegmental and segmental autonomic disorders in the development of muscular syndrome of rheumatoid arthritis].
AID: To investigate the role of suprasegmental (anxious disorders) and segmental vegetative disorders (dysfunction of neuromuscular transmission and cutaneous microcirculation) in pathogenesis of muscular syndrome of rheumatoid arthritis (RA). MMPI, Beck's questionnaire, Spilberger's test were used in experimental-psychological assessment of RA patients' personality profile. A total of 60 healthy subjects and 340 RA patients were examined. Segmental vegetative disorders were studied by excitability of hand muscles (Prognoz-6 device) and cutaneous microcirculation (doppler flowmetry). RA patients showed suprasegmental disorders manifesting as frequent anxious disturbances (95%). Dysfunction of the segmental part of the autonomic nervous system was represented by various changes in excitability of hand muscles in response to weak currents, by abnormal regulation of cutaneous microcirculation. Neuromuscular and microcirculation disturbances participate in development of RA muscular syndrome. This is confirmed by a positive correlation coefficient. There was also a negative correlation between suprasegmental and segmental disorders.


Lower placental 25-hydroxyvitamin D3 (25(OH)D3) and higher placental CYP27B1 and 25(OH)D3 ratio in preterm birth.
Neonatal mortality rates in Indonesia are still at an alarming rate, with preterm birth as one of the causes. Nutritional deficiencies such as low level of vitamin D is suspected to be the risk factors of preterm birth but still a little knowledge about it. Vitamin D metabolism includes 25-hydroxyvitamin D3 (25(OH)D3) and 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), as the inactive and active form, with the help of 1α-hydroxylase (CYP27B1) enzyme. Our study aims to determine the differences of 25(OH)D3, 1,25(OH)2D3 and CYP27B1 enzyme in term and preterm birth. A cross-sectional study was performed in Cipto Mangunkusumo National General Hospital, Jakarta, Indonesia, in January-June 2017. The blood sample was taken soon after delivery, to examine maternal 25(OH)D3 and 1,25(OH)2D3 in serum and tissue placenta, as well as placental CYP27B1 enzyme. Statistical analysis using SPPS version 20 was used to find significances. There were a total of sixty subjects in this study, with term-preterm birth group ratio 1:1. We found that placental 25(OH)D3 was significantly low (P = 0⋅001), and CYP27B1/25(OH)D3 ratio was high in preterm birth. Also, there were significant negative correlations found in CYP27B1 level and both placental 25(OH)D3 (r 0⋅481, P < 0⋅001) and 1,25(OH)2D3 (r -0⋅365, P = 0⋅004) levels. Our study concludes that preterm birth showed lower placental 25(OH)D3 status, and higher CYP27B1/25(OH)D3 ratio compared to term pregnancy.


A Novel Perspective on the ApoM-S1P Axis, Highlighting the Metabolism of ApoM and Its Role in Liver Fibrosis and Neuroinflammation.
Hepatocytes, renal proximal tubule cells as well as the highly specialized endothelium of the blood brain barrier (BBB) express and secrete apolipoprotein M (apoM). ApoM is a typical lipocalin containing a hydrophobic binding pocket predominantly carrying Sphingosine-1-Phosphate (S1P). The small signaling molecule S1P is associated with several physiological as well as pathological pathways whereas the role of apoM is less explored. Hepatic apoM acts as a chaperone to transport S1P through the circulation and kidney derived apoM seems to play a role in S1P recovery to prevent urinal loss. Finally, polarized endothelial cells constituting the lining of the BBB express apoM and secrete the protein to the brain as well as to the blood compartment. The review will provide novel insights on apoM and S1P, and its role in hepatic fibrosis, neuroinflammation and BBB integrity.


Nucleic Acid Aptamers as Potential Therapeutic and Diagnostic Agents for Lymphoma.
Lymphomas are cancers that arise from white blood cells and usually present as solid tumors. Treatment of lymphoma often involves chemotherapy, and can also include radiotherapy and/or bone marrow transplantation. There is an un-questioned need for more effective therapies and diagnostic tool for lymphoma. Aptamers are single stranded DNA or RNA oligonucleotides whose three-dimensional structures are dictated by their sequences. The immense diversity in function and structure of nucleic acids enable numerous aptamers to be generated through an iterative in vitro selection technique known as Systematic Evolution of Ligands by EXponential enrichment (SELEX). Aptamers have several biochemical properties that make them attractive tools for use as potential diagnostic and pharmacologic agents. Isolated aptamers may directly inhibit the function of target proteins, or they can also be formulated for use as delivery agents for other therapeutic or imaging cargoes. More complex aptamer identification methods, using whole cancer cells (Cell-SELEX), may identify novel targets and aptamers to affect them. This review focuses on recent advances in the use of nucleic acid aptamers as diagnostic and therapeutic agents and as targeted delivery carriers that are relevant to lymphoma. Some representative examples are also discussed.


Responses to bright light exposure in individuals with binge-spectrum eating disorders characterized by high dietary restraint and negative affect.
Circadian rhythm disruptions are associated with binge eating, can be causal of negative mood, and may be corrected with bright light exposure. A subtype of individuals with binge-spectrum eating disorders are characterized by combined high dietary restraint and negative affect. These individuals have higher eating disorder psychopathology and poorer treatment response. We aimed to test the targeted effects of morning bright light exposure on individuals with binge-spectrum eating disorders, hypothesizing significant reductions in binge eating for those characterized by high dietary restraint and negative affect. Participants (N = 34 females with binge-eating disorder and bulimia nervosa) used a morning bright light and normal light for 10 consecutive days each, in randomized order. They completed the Change in Eating Disorder Symptoms (CHEDS) scale at baseline, day 12 (when they switched lamps), and day 22. We conducted moderation analyses, clustering data by person, controlling for order, and examining the effect of light condition on binge eating according to baseline restraint and negative affect. At high levels of combined dietary restraint and negative affect, participants experienced a reduction in binge eating and food preoccupation following exposure to morning bright light. There were no changes in restrictive eating, body preoccupation, body dissatisfaction, or body checking following exposure to morning bright light for these individuals. These findings suggest that morning bright light may be a useful adjunct to empirically supported eating disorder treatments that target binge eating, especially for individuals characterized by the difficult to treat restraint/negative affect subtype. At high levels of combined dietary restraint and negative affect, participants with binge-spectrum eating disorders experienced a reduction in binge eating and food preoccupation following exposure to morning bright light. These findings suggest that morning bright light may be a useful adjunct to empirically supported eating disorder treatments that target binge eating, especially for individuals characterized by the difficult-to-treat restraint/negative affect subtype.


Use of physical and intellectual activities and socialization in the management of cognitive decline of aging and in dementia: a review.
Lifestyle nonpharmacological interventions can have a deep effect on cognitive aging. We have reviewed the available literature on the effectiveness of physical activity, intellectual stimulation, and socialization on the incidence of dementia and on the course of dementia itself. Even though physical activity appears to be beneficial in both delaying dementia onset and in the course of the disease, more research is needed before intellectual stimulation and socialization can be considered as treatments and prevention of the disease. Through our paper, we found that all three nonpharmacological treatments provide benefits to cognition and overall well-being in patients with age-related cognitive impairments. These interventions may be beneficial in the management of dementia.


Molecular mechanisms contributing to TARP regulation of channel conductance and polyamine block of calcium-permeable AMPA receptors.
Many properties of fast synaptic transmission in the brain are influenced by transmembrane AMPAR regulatory proteins (TARPs) that modulate the pharmacology and gating of AMPA-type glutamate receptors (AMPARs). Although much is known about TARP influence on AMPAR pharmacology and kinetics through their modulation of the extracellular ligand-binding domain (LBD), less is known about their regulation of the ion channel region. TARP-induced modifications in AMPAR channel behavior include increased single-channel conductance and weakened block of calcium-permeable AMPARs (CP-AMPARs) by endogenous intracellular polyamines. To investigate how TARPs modify ion flux and channel block, we examined the action of γ-2 (stargazin) on GluA1 and GluA4 CP-AMPARs. First, we compared the permeation of organic cations of different sizes. We found that γ-2 increased the permeability of several cations but not the estimated AMPAR pore size, suggesting that TARP-induced relief of polyamine block does not reflect altered pore diameter. Second, to determine whether residues in the TARP intracellular C-tail regulate polyamine block and channel conductance, we examined various γ-2 C-tail mutants. We identified the membrane proximal region of the C terminus as crucial for full TARP-attenuation of polyamine block, whereas complete deletion of the C-tail markedly enhanced the TARP-induced increase in channel conductance; thus, the TARP C-tail influences ion permeation. Third, we identified a site in the pore-lining region of the AMPAR, close to its Q/R site, that is crucial in determining the TARP-induced changes in single-channel conductance. This conserved residue represents a site of TARP action, independent of the AMPAR LBD.


The Potential Mechanism of HDAC1-Catalyzed Histone Crotonylation of Caspase-1 in Nonsmall Cell Lung Cancer.
Nonsmall cell lung cancer (NSCLC) is a predominant subtype of lung cancer and accounts for over 80% of all lung cancer cases. The resistance to pemetrexed (PEM) is frequently occurred and severely affects the NSCLC therapy. Proteomic analysis of histones indicated that the histone deacetylase 1 (HDAC1) complex could hydrolyze lysine crotonylation on histone3 (H3K18cr), affecting epigenetic regulation in cancers. However, the effect of HDAC1-mediated H3K18cr on the PEM resistance of NSCLC is still unclear. Here, we aimed to explore the function of HDAC1-mediated H3K18cr in NSCLC PEM resistance. The expression of HDAC1 was upregulated in clinical NSCLC tissues and cell lines and correlated with the poor prognosis of NSCLC samples. We constructed the PEM-resistant NSCLC cell lines, and the depletion of HDAC1 remarkably reduced the viability of the cells. The proliferation of PEM-resistant NSCLC cells was decreased by HDAC1 knockdown, and the IC50 of PEM was repressed by the silencing of HDAC1 in the cells. Mechanically, we identified the enrichment of HDAC1 on the promoter of caspase-1 in PEM-resistant NSCLC cells. The depletion of HDAC1 inhibited the enrichment of histone H3K18cr and RNA polymerase II (RNA pol II) on the caspase-1 promoter in the cells. The expression of caspase-1 was suppressed by HDAC1 knockdown. The knockdown of HDAC1 reduced proliferation of PEM-resistant NSCLC cells, in which caspase-1 or GSDMD depletion reversed the effect. Clinically, the HDAC1 expression was negatively associated with caspase-1 and GSDMD in clinical NSCLC tissues, while caspase-1 and GSDMD expression was positively correlated in the samples. Therefore, we concluded that HDAC1-catalyzed histone crotonylation of caspase-1 modulates PEM sensitivity of NSCLC by targeting GSDMD.


C-terminal sequence analysis of peptides and proteins using carboxypeptidases and mass spectrometry after derivatization of Lys and Cys residues.
C-Terminal sequence analysis of peptides and proteins using carboxypeptidase digestion in combination with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is convenient for protein and peptide characterization. After a short digestion, a sequence up to 20 residues can be identified, but the total number depends on the individual sequence. Due to the accuracy limits of the MALDI time-of-flight arrangement, the assignment of several residues with close mass values, including Lys/Glx, may remain ambiguous. We have used derivatization of lysine residues by guanidination to overcome the problem of Lys identification. The reaction is rapid and specific and results in full derivatization. In the case of Cys-containing peptides, problems arise from the fact that carboxypeptidases Y and P do not cleave peptides that contain nonderivatized cystine, cysteic acid, or (carboxymethyl)cysteine. Successful identification of Cys residues within the sequence is instead achieved by conversion of Cys to 4-thialaminine by (trimethylamino)-ethylation. The two derivatizations of Lys and Cys side chains provide opportunities for proton attachment and therefore facilitate the analysis by MALDI-MS. This C-terminal sequence analysis method is also useful for large proteins after fragmentation with specific enzymes.


Factors influencing dyslipidemia in statin-treated patients in Lebanon and Jordan: results of the Dyslipidemia International Study.
Cardiovascular disease is the leading cause of death and disability worldwide. Therefore, as part of the Dyslipidemia International Study (DYSIS), we have analyzed the prevalence of lipid abnormalities and risk factors for dyslipidemia in statin-treated patients in Lebanon and Jordan. This cross-sectional, multicenter study enrolled 617 patients at 13 hospitals in Lebanon and Jordan. Patients were at least 45 years old and had been treated with statins for at least 3 months. Multivariate logistic regression analysis was used to determine patient characteristics contributing to dyslipidemia during statin therapy. Our findings indicated that 55.9% of statin-treated patients (mean age 60.3 years, 47% female) in Lebanon and Jordan did not achieve goal levels for low-density lipoprotein cholesterol which were dependent on Systematic Coronary Risk Evaluation (SCORE) risk, and 70% of patients (76% men and 63.3% of women) were at very high cardiovascular risk. Low-density lipoprotein cholesterol goals were not achieved in 67.2% of those with very high cardiovascular risk. The most commonly prescribed statin was atorvastatin (44.6%), followed by simvastatin (27.7%), rosuvastatin (21.2%), fluvastatin (3.3%), pravastatin (3%), and lovastatin (0.2%). Approximately half of the population was treated with a statin dose potency of 4, equaling 40 mg of simvastatin. In Lebanon and Jordan, the strongest independent associations with low-density lipoprotein cholesterol not at goal were current smoking (odds ratio [OR] 1.96; 95% confidence [CI] 1.25-3.08), diabetes mellitus (OR 2.53; 95% CI 1.70-3.77), and ischemic heart disease (OR 2.26; 95% CI 1.45-3.53), while alcohol consumption was associated with reduced risk (OR 0.12; 95% CI 0.03-0.57). We observed that many patients in Lebanon and Jordan experienced persistent dyslipidemia during statin treatment, supporting the notion that novel lipid-lowering strategies need to be developed. Also, social programs aimed at combating the extremely high rates of tobacco use and obesity in Lebanon and Jordan are critical for combating cardiovascular disease in these countries.


Japanese cases of type 1 thanatophoric dysplasia exclusively carry a C to T transition at nucleotide 742 of the fibroblast growth factor receptor 3 gene.
Type I thanatophoric dysplasia (TD) is typically a lethal neonatal dwarfism, but a limited number of cases of type I TD cases survive more than one year, suggesting genetic heterogeneity. In this study, we analyzed the fibroblast growth factor receptor 3 (FGFR3) gene in 5 Japanese cases of type I TD with clinical symptoms ranging from lethal to long-survival. In every case, nucleotide sequence analysis of cDNA revealed a C to T transition at nucleotide 742 (C742T) in one allele of the FGFR3 gene, suggesting that type I TD is a rather homogeneous genetic condition, irrespective of clinical course. No association was found between C742T and C882T, although both nucleotides changes were from CpG dinucleotide in a near location.


Implementing malaria control in South Africa, Eswatini and southern Mozambique during the COVID-19 pandemic.
The COVID-19 pandemic has strained healthcare delivery systems in a number of southern African countries. Despite this, it is imperative that malaria control and elimination activities continue, especially to reduce as far as possible the number and rate of hospitalisations caused by malaria. The implementation of enhanced malaria control/elimination activities in the context of COVID-19 requires measures to protect healthcare workers and the communities they serve. The aim of this review is therefore to present innovative ideas for the timely implementation of malaria control without increasing the risk of COVID-19 to healthcare workers and communities. Specific recommendations for parasite and vector surveillance, diagnosis, case management, mosquito vector control and community outreach and sensitisation are given.


2006 Philip Kittredge Memorial Lecture. What to do when protocols fail.
Though advances in medical science have created improved therapies, often these are not widely provided throughout the health-care system. Also, there is growing recognition of the lack of safety in health-care delivery. The development of evidence-based, best practice, national guidelines has been encouraged to reduce unnecessary variation in care and for improving quality. Adoption of guidelines through local protocols has been disappointingly slow. This paper explores the parallel developments in safety and quality-of-care assessment, evidence-based medicine, guideline creation, and how development of national and international quality-improvement campaigns are promoting rapid change in care delivery processes. I discuss how this new opportunity can improve the quality of respiratory care and enhance the adoption of respiratory care protocols.


Transcriptional restriction of human immunodeficiency virus type 1 gene expression in undifferentiated primary monocytes.
Monocytes are critical precursors of dendritic cells and macrophages, which play an important role in the pathogenesis of human immunodeficiency virus type 1 (HIV-1). HIV-1 postentry infection is blocked in undifferentiated monocytes in vitro, while the underlying mechanisms are not fully understood. HIV-1 Tat-mediated transactivation of the viral long terminal repeat (LTR) promoter is essential for HIV-1 transcription. Two critical cellular cofactors of HIV-1 Tat, cyclin T1 (CycT1) and cyclin-dependent kinase 9 (CDK9), are required for LTR-directed HIV-1 transcription. In addition to the previously identified restrictions in early viral life cycle, we find that HIV-1 gene expression is impaired in undifferentiated primary monocytes. Transfection of monocytes by nucleofection with HIV-1 proviral DNA could not produce infectious HIV-1. The lack of Tat transactivation of the LTR promoter correlated with the impaired HIV-1 gene expression in monocytes. Interestingly, heterokaryons between primary monocytes and a human embryonic kidney cell line restored Tat transactivation of LTR, suggesting that monocytes lack cellular factors required for Tat transactivation. CycT1 protein was undetectable in freshly isolated monocytes and induced in monocyte-differentiated macrophages, while the expression of CDK9 remained constant. Transient expression of CycT1 in undifferentiated monocytes could not rescue Tat transactivation, suggesting that CycT1 is not the only limiting factor of HIV-1 infection in monocytes. Furthermore, monocyte differentiation into macrophages appeared to enhance the phosphorylation of CDK9, which correlated with significantly increased HIV-1 infection in macrophages. Our results provide new insights into HIV-1 infection and regulation in primary monocytes and viral pathogenesis.


Effect of chlorhexidine digluconate on antimicrobial activity, cell viability and physicochemical properties of three endodontic sealers.
Assess the biological and physicochemical properties of AH Plus, BioRoot RCS and Pulp Canal Sealer (PCS) leachates with and without chlorhexidine (CHX). The sealers were studied in no contact and 1-minute contact with CHX. For biological properties (antibacterial activity and cytotoxicity), leachates were formed in saline of freshly mixed, 1-, 7- and 28 days set sealers. The antibacterial properties of sealer leachates were investigated for planktonic and biofilm growth of E. faecalis, S. mutans, S.epidermidis and S.aureus. The 3-(4,5 dimethylthiazolyl-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to evaluate murine fibroblast cell viability after exposure to the leachates. The physical properties (water uptake, sorption, solubility, porosity, surface characteristics) of sealers and the pH of the immersion liquid (saline or distilled water) were also assessed over a 28-days period. CHX improved the antibacterial properties of the sealer leachates and reduced cell viability for all sealer leachates, except for freshly mixed PCS. BioRoot RCS leachates presented the highest antibacterial properties and cell viability with and without CHX contact. PCS was the material most affected by CHX in terms of physical properties, whereas for AH Plus, solubility was increased. CHX did not affect the physical properties of BioRoot RCS, except for solubility that was decreased. CHX contact did not change sealers' alkalinity in distilled water whereas it increased it for AH Plus and BioRoot RCS in saline. CHX improved the antibacterial efficacy of sealer leachates and either compromised or did not affect cell viability. CHX affected to various extent sealers' physicochemical properties.


Rab25 influences functional Cav1.2 channel surface expression in arterial smooth muscle cells.
Plasma membrane-localized CaV1.2 channels are the primary calcium (Ca(2+)) influx pathway in arterial smooth muscle cells (myocytes). CaV1.2 channels regulate several cellular functions, including contractility and gene expression, but the trafficking pathways that control the surface expression of these proteins are unclear. Similarly, expression and physiological functions of small Rab GTPases, proteins that control vesicular trafficking in arterial myocytes, are poorly understood. Here, we investigated Rab proteins that control functional surface abundance of CaV1.2 channels in cerebral artery myocytes. Western blotting indicated that Rab25, a GTPase previously associated with apical recycling endosomes, is expressed in cerebral artery myocytes. Immunofluorescence Förster resonance energy transfer (immunoFRET) microscopy demonstrated that Rab25 locates in close spatial proximity to CaV1.2 channels in myocytes. Rab25 knockdown using siRNA reduced CaV1.2 surface and intracellular abundance in arteries, as determined using arterial biotinylation. In contrast, CaV1.2 was not located nearby Rab11A or Rab4 and CaV1.2 protein was unaltered by Rab11A or Rab4A knockdown. Rab25 knockdown resulted in CaV1.2 degradation by a mechanism involving both lysosomal and proteasomal pathways and reduced whole cell CaV1.2 current density but did not alter voltage dependence of current activation or inactivation in isolated myocytes. Rab25 knockdown also inhibited depolarization (20-60 mM K(+)) and pressure-induced vasoconstriction (myogenic tone) in cerebral arteries. These data indicate that Rab25 is expressed in arterial myocytes where it promotes surface expression of CaV1.2 channels to control pressure- and depolarization-induced vasoconstriction.


An analysis of the effects of apomorphine and haloperidol on the release of [3H]-dopamine and [3H]-noradrenaline from rabbit brain slices.
The effects of apomorphine and haloperidol on the K+-induced release of [3H]-dopamine from striatal slices and [3H]-noradrenaline from cerebellar slices were examined. Apomorphine (0.01-1 microM) reduced the K+-induced release of [3H]-dopamine from striatal slices, leaving the K+-induced release of [3H]-noradrenaline from cerebellar slices unaffected. A higher concentration (10 microM) of apomorphine increased significantly the K+-induced release of [3H]-noradrenaline from cerebellar slices. Haloperidol (0.01-1 nM) increased the K+-induced release of [3H]-dopamine from striatal slices, leaving the K+-induced release of [3H]-noradrenaline unaffected. Haloperidol in higher concentrations (10-100nM) increased significantly the K+-induced release of [3H]-noradrenaline from cerebellar slices. Both compounds inhibited the uptake of [3H]-dopamine into striatal slices and [3H] noradrenaline into cerebellar slices. In low concentrations both compounds showed selectivity for dopaminergic receptor sites, apomorphine stimulating and haloperidol inhibiting pre-synaptic dopamine receptors. Higher concentrations of these compounds showed a non-selective action.


How do children with attention deficit hyperactivity disorder interact in a clinical dental examination? A video analysis.
Attention deficit hyperactivity disorder (ADHD) is currently the most common behavioural disorder in school-age children. The aim of this study was to perform a detailed analysis of behavioural interactions between the dentist and the child patient with ADHD. All children born in 1991 (n = 555) in one Swedish municipality were screened for attention and learning problems, and assessed for ADHD. Twenty-two children with ADHD, and a control group of 47 children without attention and learning problems, were included in the study. The dental recall visit was recorded on video. The interaction between the dentist and the child was analysed in detail and scored as verbal and non-verbal initiatives and responses. Compared to the children in the control group, the children with ADHD made significantly more initiatives, especially initiatives that did not focus on the examination or the dentist. The children with ADHD had fewer verbal responses and more missing responses. In conclusion, the problems in communication resulted in less two-way communication between the dentist and the children with ADHD than the interaction between the dentist and the children in the control group. The children with ADHD had particular difficulties staying focused on the examination.


Non-major histocompatibility complex-restricted cytotoxicity of bovine coronavirus-infected target cells mediated by bovine intestinal intraepithelial leukocytes.
Non-specific cellular mechanisms of defence against intestinal virus infections of cattle were investigated using bovine coronavirus (BCV) as a representative enteric virus. Since BCV infection is limited to the epithelial cells of the intestinal tract, defence mechanisms must be capable of acting at that site to be effective. Therefore, the intraepithelial leukocyte (IEL) population of the intestinal mucosa was chosen for initial study. Treatment of intestinal samples with DTT and EDTA in calcium- and magnesium-free buffers allowed recovery of viable IEL populations appropriate for further functional assessment. Studies of IELs isolated from neonatal calves revealed that non-major histocompatibility complex (MHC)-restricted cytotoxicity of BCV-infected target cells was more prevalent in calves with concurrent virus infection, suggesting in vivo activation of the cytotoxic response. Peripheral blood mononuclear cells from the same calves did not mediate cytotoxicity, emphasizing the difference in function of lymphocytes isolated from different anatomical sites. IELs from normal adult animals rarely showed spontaneous non-MHC-restricted cytotoxicity. However, interleukin-2 (IL-2) was a potent activator of IEL cytotoxicity in vitro, enhancing the killing of BCV-infected target cells after just 18 h of treatment. Incubation of IELs with interferon-gamma and tumour necrosis factor (TNF) did not induce cytotoxic activity, but TNF could augment the levels of IL-2-induced cytotoxicity. Although further analysis of the cytotoxic effector cells present in the intestinal epithelium is required, the present study indicates that the IEL population may play a role in enteric antiviral activity.


Commentary: till we have faces--on humiliation.
My commentary takes up two points raised by Drs. Torres and Bergner: first, the social context in which humiliation takes place and the way such behavior is normalized; and second, the question of face, and in this context, the loss of face and its impact on identity. For me, the key concern in relation to humiliation is not so much loss of entitlement status, but a loss of identity. Humiliation is a fundamental attack on narrative identity, but I will make the (perhaps uncomfortable) suggestion that not everyone is humiliate-able and that, to some extent, one may be able to choose the extent of one's injuries through loss of face.


Tactile hypersensitivity and GABA concentration in the sensorimotor cortex of adults with autism.
Sensory hypersensitivity is frequently encountered in autism spectrum disorder (ASD). Gamma-aminobutyric acid (GABA) has been hypothesized to play a role in tactile hypersensitivity. The aim of the present study was twofold. First, as a study showed that children with ASD have decreased GABA concentrations in the sensorimotor cortex, we aimed at determining whether the GABA reduction remained in adults with ASD. For this purpose, we used magnetic resonance spectroscopy to measure GABA concentration in the sensorimotor cortex of neurotypical adults (n = 19) and ASD adults (n = 18). Second, we aimed at characterizing correlations between GABA concentration and tactile hypersensitivity in ASD. GABA concentration in the sensorimotor cortex of adults with ASD was lower than in neurotypical adults (decrease by 17%). Interestingly, GABA concentrations were positively correlated with self-reported tactile hypersensitivity in adults with ASD (r = 0.50, P = 0.01), but not in neurotypical adults. In addition, GABA concentrations were negatively correlated with the intra-individual variation during threshold measurement, both in neurotypical adults (r = -0.47, P = 0.04) and in adults with ASD (r = -0.59, P = 0.01). In other words, in both groups, the higher the GABA level, the more precise the tactile sensation. These results highlight the key role of GABA in tactile sensitivity, and suggest that atypical GABA modulation contributes to tactile hypersensitivity in ASD. We discuss the hypothesis that hypersensitivity in ASD could be due to suboptimal predictions about sensations. Autism Research 2019, 12: 562-575. © 2019 International Society for Autism Research, Wiley Periodicals, Inc. LAY SUMMARY: People with autism spectrum disorder (ASD) often experience tactile hypersensitivity. Here, our goal was to highlight a link between tactile hypersensitivity and the concentration of gamma-aminobutyric acid (GABA) (an inhibitory neurotransmitter) in the brain of adults with ASD. Indeed, self-reported hypersensitivity correlated with reduced GABA levels in brain areas processing touch. Our study suggests that this neurotransmitter may play a key role in tactile hypersensitivity in autism.


Development of a novel proteomic approach for the detection of transitional cell carcinoma of the bladder in urine.
Development of noninvasive methods for the diagnosis of transitional cell carcinoma (TCC) of the bladder remains a challenge. A ProteinChip technology (surface enhanced laser desorption/ionization time of flight mass spectrometry) has recently been developed to facilitate protein profiling of biological mixtures. This report describes an exploratory study of this technology as a TCC diagnostic tool. Ninety-four urine samples from patients with TCC, patients with other urogenital diseases, and healthy donors were analyzed. Multiple protein changes were reproducibly detected in the TCC group, including five potential novel TCC biomarkers and seven protein clusters (mass range, 3.3 to 133 kd). One of the TCC biomarkers (3.4 kd) was also detected in bladder cancer cells procured from bladder barbotage and was identified as defensin. The TCC detection rates provided by the individual markers ranged from 43 to 70% and specificities from 70 to 86%. Combination of the protein biomarkers and clusters, increased significantly the sensitivity for detecting TCC to 87% with a specificity of 66%. Interestingly, this combinatorial approach provided sensitivity of 78% for detecting low-grade TCC compared to only 33% of voided urine or bladder-washing cytology. Collectively these results support the potential of this proteomic approach for the development of a highly sensitive urinary TCC diagnostic test.


Erdheim-Chester Disease Due to a Novel Internal Duplication of NRAS: Response to Targeted Therapy with Cobimetinib.
Histiocytoses encompass a group of exceptionally rare disorders characterized by the abnormal infiltration of tissues by histocytes. Among these, Erdheim-Chester disease (ECD) stands out as a multisystem histiocytosis that typically affects bones and various other tissues. Historically, the treatment of ECD has been challenging. However, recent breakthroughs in our understanding, particularly the discovery of somatic mutations in the RAS-MAPK pathway, have opened new opportunities for targeted therapy in a significant subset of patients with ECD and other histiocytoses. In this report, we present the case of a patient with ECD harboring a previously unidentified microduplication in the NRAS gene in a small fraction of skin cells. This discovery played a pivotal role in tailoring an effective therapeutic approach involving kinase inhibitors downstream of NRAS. This case underscores the crucial role of deep sequencing of tissue samples in ECD, enabling the delivery of personalized targeted therapy to patients.