Expression and tissue distribution of astacin-like squid metalloprotease (ALSM).
Astacin metalloprotease family members function in a wide variety of biologic events, including cell differentiation and morphogenesis during embryonic development and adult tissue differentiation. We previously isolated and characterized an astacin-like squid metalloprotease (ALSM). To elucidate the embryonic expression of ALSM, we performed immunohistochemical analysis with specific antibodies and examined the expression profiles of ALSM isoforms by in situ hybridization analysis. Tissue distribution and expression were also examined in adult spear squid. mRNA expression of ALSM isoforms I and III was first detected in newly hatched squid and was restricted to the liver. No mRNA signals were detected in other tissues even in adult squids. At the protein level, both isoforms were prominent in the liver of embryos and later in digestive organs of adult squid. Both isoforms were also detected in muscle tissues, including mantle and tentacle muscle. Staining for ALSM III was also identified in the iris and in tissues near the eye in squid embryos. However, no reactive bands were detected by immunoblotting of adult squid eyes. Thus, ALSM is initially expressed at the late stage of embryogenesis in spear squid, and expression is restricted to the liver. Thereafter, ALSM isoforms function in various tissues in an isoform-dependent manner.


Flat panel detector computed tomography for the interaction between contrast-enhanced thrombi and stent retrievers in stroke therapy: a pilot study.
The purpose of this study was to create virtual 3D views of the interaction between contrast-enhanced thrombi and three different types of stent retrievers. Artificial thrombi with and without contrast agent were created and introduced into a silicone tube with saline solution. The stent retrievers (Aperio (A), Solitaire FR (B), Revive (C)) were released around the thrombi. For each retriever, two sets (0 min, 5 min) of flat panel computed tomography (CT) data were acquired on a Siemens Axiom Artis Zee biplane angiography system and reconstructed using syngo InSpace 3D software. The filaments of all three stent retrievers were displaced by the thrombus immediately after deployment. Another study series after 5 min showed further expansion of the stents into the thrombus. A tends to appose best around obstacles. A and B allow cushion-like bulging of clot material into the stent lumen. C showed only moderate curvilinear bulging and a minor degree of interaction in between stent filaments and thrombus. Flexible design allows better apposition of the device to the surface of the thrombus. A relatively strong longitudinal structure combined with large gaps in between the stent filaments seems to favor migration into the clot. The main capture mechanism seems to be engagement of the clot between the crossings of stent struts.


Taste preference of Chlorella sp. algae from dairy wastewater by weaned dairy calves.
The objective of this study was to evaluate the taste preference of calves fed Chlorella sp. microalgae produced from dairy lagoon wastewater. Six Holstein dairy heifer calves that were 12 to 14 wk of age (107 ± 3.8 kg of body weight) were fed 0 (control), 30, or 60 g of Chlorella sp. daily, and all calves were fed all treatments in a sequential elimination study. For the 7-d experiment, d 1 to 2 were for diet adaptation and d 3 to 4 were for data collection. During the final 3 d, the primary consumed treatment was removed to determine the second preferred treatment. Calves were ranked for total intake from the consumption of all treatments. The microalgae product used in this study was isolated from dairy wastewater lagoon, and microalgae biomass was produced using outdoor hanging bag bioreactors with Chlorella sp. to recycle the wastewater. The biomass was sterilized and kept frozen at -4°C until fed to calves. Calves were housed individually in hutches with outdoor access under solar panels, with free-choice water. Calves consumed more dry matter from control calf starter (3.4 kg/d) than from the starter with 30 g (2.42 kg/d) or 60 g (1.56 kg/d) of microalgae during the first 2-d period. During the second 2-d period (d 3 and 4), dry matter intake was reduced for the 60 g/d microalgae starter compared with the control and 30 g/d microalgae starters. Five of 6 calves in this study always ranked the control treatment first when given a choice and ranked the 30 g of microalgae starter second choice. Results indicated that microalgae may be added to calf starter; however, calves preferred calf starter without microalgae.


The impact of preprinted prescription forms on medication prescribing errors in an ophthalmology clinic in northeast Thailand: a non-randomised interventional study.
To understand the incidence and types of medication prescribing errors in a low resource setting ophthalmology clinic and to determine the impact of a preprinted prescription based on the hospital formulary (FormularyScript) on medication prescribing errors. Non-randomised interventional study. Ophthalmology clinic in a teaching hospital in northeast Thailand. 4349 handwritten prescriptions collected from October 2009 to December 2009, and 4146 FormularyScripts collected from February 2010 to May 2010. All prescriptions from the handwritten and FormularyScript groups were analysed for medication error rates by types (legibility, ambiguous, incomplete, abbreviation and accuracy) and subtypes (drug name, strength, which eye, route and dispensed amount). Comparison of error rates in the two groups showed a 10-fold reduction in the overall error rate using FormularyScript (32.9%-3.5%, p<0.001). FormularyScripts were associated with statistically significant (p<0.001) decreases in the following error types: legibility (16.1%-0.1%), incomplete (16.1%-0.1%) and abbreviation (3.1%-0.3%). There was no statistically significant change in accuracy errors (0.8%-0.6%, p=0.21). Ambiguous errors increased with FormularyScripts (0.6%-2.5%, p<0.001), likely due to the introduction of new ways to make errors. Decreases were seen in all legibility, abbreviation and accuracy error subtypes, and four out of six incomplete error subtypes. There were statistically significant increases in both ambiguous error subtypes: which eye (0.3%-2.5%, p<0.001) and drug name (0.3%-0.6%, p=0.03). In our study population, outpatient medication prescribing errors were common and primarily due to legibility and incomplete error types. A preprinted prescription form has the potential to decrease medication prescribing errors related to legibility, incomplete prescribing information and use of unacceptable abbreviations without changing the overall rate of accuracy errors. However, new error types can occur.


Model for calcium dependent oscillatory growth in pollen tubes.
Experiments have shown that pollen tubes grow in an oscillatory mode, the mechanism of which is poorly understood. We propose a theoretical growth model of pollen tubes exhibiting such oscillatory behaviour. The pollen tube and the surrounding medium are represented by two immiscible fluids separated by an interface. The physical variables are pressure, surface tension, density and viscosity, which depend on relevant biological quantities, namely calcium concentration and thickness of the cell wall. The essential features generally believed to control oscillating growth are included in the model, namely a turgor pressure, a viscous cell wall which yields under pressure, stretch-activated calcium channels which transport calcium ions into the cytoplasm and an exocytosis rate dependent on the cytosolic calcium concentration in the apex of the cell. We find that a calcium dependent vesicle recycling mechanism is necessary to obtain an oscillating growth rate in our model. We study the variation in the frequency of the growth rate by changing the extracellular calcium concentration and the density of ion channels in the membrane. We compare the predictions of our model with experimental data on the frequency of oscillation versus growth speed, calcium concentration and density of calcium channels.


The pulmonary side of reflux disease: from heartburn to lung fibrosis.
Gastroesophageal reflux disease (GERD) is the most prevalent gastrointestinal disorder in the USA. Heartburn is the symptom most commonly associated with this disease, and the highly commercialized medical treatment directed toward relief of this symptom represents a 10-billion-dollar-per-year industry. Unfortunately, there is often little awareness that GERD can be potentially a lethal disease as it can cause esophageal cancer. Furthermore, there is even less awareness about the relationship between GERD and respiratory disorders with the potential for severe morbidity and even mortality.


Annotated checklist of the Blennidus subgenus Agraphoderus species from Peru with description of B. bombonensis n. sp. and synonymic notes (Coleoptera: Carabidae: Pterostichinae).
Forty species belonging to the subgenus Agraphoderus of Blennidus have been recorded so far from Peru. An annotated checklist is provided with information about their type locality, distribution and habitat. The nomenclature of each species is also provided, together with some notes on their systematic status. Blennidus bombonensis n. sp. from Cerro de Pasco is described; Blennidus pseudangularis nomen novum for Ogmopleura angularis Straneo, 1993 (nec Straneo, 1985) is proposed and the following synonymies are stated: Blennidus pseudangularis Allegro & Giachino, nomen novum = Blennidus rectangulus (Straneo, 1993) syn. nov.; Ogmopleura minor Straneo, 1993 = Blennidus rectangulus (Straneo, 1993) syn. nov. Type specimens of most species are illustrated, as well as male genitalia. Finally, a revised key to all the Agraphoderus species from Peru is provided.


Sensitive immunoassay of human chorionic gonadotrophin based on multi-walled carbon nanotube-chitosan matrix.
A novel amperometric immunosensor for human chorionic gonadotropin (HCG) assay has been fabricated through incorporating toluidine blue (TB) and hemoglobin (Hb) on the multiwall carbon nanotube (MWNT)-chitosan (CS) modified glassy carbon electrode, followed by electrostatic adsorption of a conducting gold nanoparticles (nanogold) film as sensing interface. The MWNT-CS matrix provided a congenial microenvironment for the immobilization of biomolecules and promoted the electron transfer to enhance the sensitivity of the immunosensor. Due to the strong electrocatalytic properties of Hb and MWNT toward H(2)O(2), the Hb and MWNT significantly amplified the current signal of the antigen-antibody reaction. The immobilized toluidine blue as an electron transfer mediator exhibited excellent electrochemical redox property. After the immunosensor was incubated with HCG solution, the access of activity center of the Hb to toluidine blue was partly inhibited, which leaded to a linear decrease in the catalytic efficiency of the Hb to the oxidation of immobilized toluidine blue by H(2)O(2) over HCG concentration ranges from 0.8 to 500 mIU/mL. Under optimal condition, the detection limit for the HCG immunoassay was 0.3 mIU/mL estimated at a signal-to-noise ratio of 3. Moreover, the proposed immunosensor displayed a satisfactory stability and reproducibility.


Improvement of cholera toxin-catalyzed ADP-ribosylation by endogenous ADP-ribosylation factor from bovine brain provides evidence for an unchanged amount of Gs alpha in failing human myocardium.
The aim of the present study was to investigate whether or not alterations of Gs alpha can be detected with cholera toxin-induced ADP-ribosylation in myocardial membranes from patients with heart failure. Therefore, Gs alpha was radiolabeled by cholera toxin-catalzyed (32P)ADP-ribosylation with (32P)NAD as substrate. In membranes from left ventricular myocardium of six patients with dilated cardiomyopathy classified as NYHA IV and three samples from two non-failing donor hearts, labeling was too weak to allow detection of possible changes in the amount of Gs alpha. Therefore, the cytosolic small molecular weight G protein ARF (ADP-ribosylation factor), a cofactor for cholera toxin-induced ADP-ribosylation of Gs alpha, was partially purified from bovine cerebral cortex. ARF activity was quantified by its ability to enhance auto-ADP-ribosylation of cholera toxin A1-subunit. Gs alpha was identified by comparing the ADP-ribosylation patterns of myocardial membranes, membranes prepared from human leukemia (HL 60) and S 49 mouse lymphoma wild type cells (45 kDa-band present) with membranes of the Gs alpha-deficient S 49 variant cyc- (45 kDa-band missing). In the presence of ARF, specific radiolabeling of the Mr 45,000 subtype of Gs alpha was markedly enhanced. The amounts of Gs alpha as measured by cholera toxin-dependent (32P)-ADP-ribosylation in the presence of ARR were similar in failing and nonfailing human hearts. It is concluded that factors other than Gs alpha are responsible for the altered regulation of the adenylate cyclase complex in heart failure. Moreover, by enhancing cholera toxin-catalyzed ADP-ribosylation, endogenous ADP-ribosylation factor from bovine brain appears to be a useful tool to study Gs alpha even in tissues in which the labeling of Gs alpha is rather weak.


PAX8 expression in anaplastic thyroid carcinoma is less than those reported in early studies: a multi-institutional study of 182 cases using the monoclonal antibody MRQ-50.
Anaplastic thyroid carcinoma (ATC) is an aggressive malignant tumor composed of undifferentiated thyroid follicular cells. Pathological diagnosis of ATC can be challenging as the tumor may show morphological overlap with other neoplasms with anaplastic morphology. Immunohistochemical demonstration of thyroid origin facilitates the diagnosis of ATC. Previous studies using the polyclonal anti-PAX8 antibody 10336-1-AP suggested that PAX8 was the most sensitive marker, expressed in up to 80% of ATC. According to a 2018 NordiQC report, the monoclonal anti-PAX8 antibody MRQ-50 has become the most commonly used anti-PAX8 antibody worldwide. However, validation of this antibody in ATC is lacking. In this study, we recruited 182 ATC cases from seven institutions. Pathology slides were subjected to histology review. PAX8 immunohistochemistry using the MRQ-50 antibody was performed in whole tissue slides (n = 147) or tissue microarray sections (n = 35). We found PAX8 expression in 54.4% of the cases, which was significantly lower than those reported in prior studies with the polyclonal antibody. PAX8 expression was positively correlated with the presence of an epithelial pattern (63.6% vs 37.5%, p = 0.0008) and a coexisting differentiated thyroid carcinoma component (71.6% vs 44.3%, p = 0.0004), but was not associated with age, gender, specimen type, or presence of giant cell and sarcomatoid patterns. In conclusion, we demonstrated PAX8 expression using the monoclonal antibody MRQ-50 in only half of the cases in a large ATC series. Pathologists should be aware that PAX8 expression in ATC is less than those reported in early studies to avoid misdiagnosis.


Interaction of SOCS3 with NonO attenuates IL-1beta-dependent MUC8 gene expression.
The intracellular negatively regulatory mechanism which affects IL-1beta-induced MUC8 gene expression remains unclear. We found that SOCS3 overexpression suppressed IL-1beta-induced MUC8 gene expression in NCI-H292 cells, whereas silencing of SOCS3 restored IL-1beta-induced MUC8 gene expression. Sequentially activated ERK1/2, RSK1, and CREB by IL-1beta were not affected by SOCS3, indicating that SOCS3 has an independent mechanism of action. Using immunoprecipitaion and nano LC mass analysis, we found that SOCS3 bound NonO (non-POU-domain containing, octamer-binding domain protein) in the absence of IL-1beta, whereas IL-1beta treatment dissociated the direct binding of SOCS3 and NonO. A dominant-negative SOCS3 mutant (Y204F/Y221F) did not bind to NonO. Interestingly, SOCS3 overexpression dramatically suppressed MUC8 gene expression in cells transfected with wild-type or siRNA of NonO. Moreover, silencing of SOCS3 dramatically increased NonO-mediated MUC8 gene expression caused by IL-1beta compared to NonO overexpression alone, suggesting that SOCS3 acts as a suppressor by regulating the action of NonO.


Wild or Farmed Gilthead Seabream (Sparus aurata)? How To Distinguish between Them by Two-Dimensional Gel Electrophoresis.
Because the world's wild fish stocks are limited and the market demand is increasing, fish farming has become an alternative food source and a way to reduce costs for consumers. The sale of farmed as wild fish is a fraudulent practice; it is, therefore, important to find new and alternative tools that can help in the fight against fraud to protect consumers and to ensure food traceability. The proteomic profiles of farmed and wild fish differ. With this study we wanted to identify liver protein markers via two-dimensional electrophoresis that would allow us to distinguish wild from farmed gilthead seabream. The liver samples from 32 gilthead seabream, wild and farmed, were stored at -80°C before protein extraction. The samples were subjected to two-dimensional electrophoresis to detect qualitative and quantitative differences. Proteomic analysis showed a protein spot (molecular weight of ∼34 kDa and isoelectric point of ∼6.9) only in the samples from the wild gilthead seabream; liquid chromatography-tandem mass spectrometry identified the spot as ubiquitin. Ubiquitin could be a valid marker to differentiate wild from farmed gilthead seabream; it could be used to ensure continuous monitoring throughout the entire commercial chain and to fight commercial fraud.


A microwave-assisted and heteropolyacids-catalysed cyclocondensation reaction for the synthesis of 4(3H)-quinazolinones.
We have investigated a microwave-assisted synthesis of 4(3H)-quinazolinones by condensation of anthranilic acid, orthoesters (or formic acid) and substituted anilines,using Keggin-type heteropolyacids (H(3)PW(12)O(40).13H(2)O, H(4)SiW(12)O(40).13H(2)O,H(4)SiMo(12)O(40).13H(2)O or H(3)PMo(12)O(40).13H(2)O) as catalysts. We found that the the use of H(3)PW(12)O(40).13H(2)O acid coupled to microwave irradiation allows a solvent-free, rapid (approximately 13min) and high-yielding reaction.


[Epidermal aging and anti-aging strategies].
Epithelial senescence is a complex process depending on intrinsic as well as extrinsic factors (e.g., UV or IR light, tobacco smoke) and must be seen in the context of the aging process especially of the corium and the subcutis. Morphological alterations become apparent in the form of epithelial atrophy, structural changes within the basal membrane, and a decrease in cell count of melanocytes and Langerhans cells. Signs of cellular senescence are reduced proliferation of keratinocytes, cumulation of dysplastic keratinocytes, various mutations (e.g., c-Fos/c-Jun, STAT3, FoxO1), as well as multiple lipid or amino acid metabolic aberrations (e.g., production of advanced glycation endproducts). This causes functional changes within the physical (lipid deficiency, water distribution dysfunction, lack of hygroscopic substances), chemical (pH conditions, oxygen radicals), and immunological barrier. Prophylactically, barrier-protective care products, antioxidant substances (e.g., vitamin C, B3, E, polyphenols, flavonoids), sunscreen products/measurements, and retinoids are used. For correcting alterations in aged epidermis, chemical peelings (fruit acids, β-hydroxy acid, trichloroacetic acid, phenolic compounds), non-ablative (IPL, PDL, Nd:YAG) as well as ablative (CO2, Erbium-YAG) light-assisted methods are used.


AID-driven deletion causes immunoglobulin heavy chain locus suicide recombination in B cells.
Remodeling of immunoglobulin genes by activation-induced deaminase (AID) is required for affinity maturation and class-switch recombination in mature B lymphocytes. In the immunoglobulin heavy chain locus, these processes are predominantly controlled by the 3' cis-regulatory region. We now show that this region is transcribed and undergoes AID-mediated mutation and recombination around phylogenetically conserved switchlike DNA repeats. Such recombination, which we term locus suicide recombination, deletes the whole constant region gene cluster and thus stops expression of the immunoglobulin of the B cell surface, which is critical for B cell survival. The frequency of this event is approaching that of class switching and makes it a potential regulator of B cell homeostasis.


Hospital readmissions for newly discharged pediatric home mechanical ventilation patients.
Ventilator-dependent children have complex chronic conditions that put them at risk for acute illness and repeated hospitalizations. To determine the 12-month incidence of and risk factors for non-elective readmission in children with chronic respiratory failure (CRF) after initiation on home mechanical ventilation (HMV) via tracheostomy. A retrospective cohort study of 109 HMV patients initiated and followed at an university-affiliated children's hospital between 2003 and 2009. Patient characteristics are presented using descriptive statistics; generalized estimated equations are used to estimate adjusted odds ratios of select predictor variables for readmission. The 12-month incidence of non-elective readmission was 40%. Close to half of these readmissions occurred within the first 3 months post-index discharge. Pneumonia and tracheitis were the most common reasons for readmission; 64% were pulmonary- or tracheostomy-related. Most demographic and clinical patient characteristics were not statistically associated with non-elective readmissions. Although, a change in the child's management within 7 days before discharge was associated readmissions shortly after index discharge. Non-elective readmissions of newly initiated pediatric HMV patients were common and likely multifactorial. Many of these readmissions were airway-related, and some may have been potentially preventable.


Crohn's disease: the cold chain hypothesis.
A recent published hypothesis proposed that Crohn's disease was provoked by infantile exposure to micro-organisms that can survive refrigerator temperature. A case-control study was accordingly devised. The mean age at first fridge was 5.6 years amongst 88 patients with Crohn's disease, 5.5 years in 88 patients with ulcerative colitis (UC) and 7.6 years in 88 controls, but a majority of individuals had always been exposed to refrigerated food. Differences were more striking in subjects aged above the median (10.3, 10.9 and 15.0 years for Crohn's disease, UC and controls, respectively). This support for the hypothesis reached statistical significance for those with Crohn's disease compared to the controls (p=0.045).


Evidence for genetic linkage between a polymorphism in the GNAS gene and malaria in South Indian population.
The complex imprinted GNAS locus which encodes G-alpha subunit (Gαs) is involved in a number of G-protein coupled signaling pathways in eukaryotic cells. Erythrocyte invasion by Plasmodium falciparum parasites is significantly regulated by protein of GNAS gene. This study was designed to evaluate the association between single nucleotide polymorphisms (SNPs) present in GNAS locus and susceptibility to malaria. In this case control study, individuals affected by P. falciparum malaria (n=230), Plasmodium vivax malaria (n=230) and normal controls (n=230) were tested for the association of eighteen (18) known SNPs to evaluate their role in the onset of the disease. There was no significant difference in genotype frequencies of all the SNPs tested between P. falciparum and P. vivax affected individuals. However, when Bonferroni correction for multiple comparisons were performed as a control, our results demonstrated alleles and genotypes of rs7121: C>T (NC_000020.10:g.57478807C>T), a silent polymorphism situated in the exon 5, were significantly (p<0.05) associated with susceptibility to malaria in the South Indians participants. Our results demonstrate that population specific polymorphisms that exist in GNAS gene may alter the risk of occurrence of malaria.


The contextual goal dependent attentional flexibility (CoGoDAF) framework: A new approach to attention bias in depression.
Successful adaptation to the environment requires attentional prioritization of emotional information relevant to the current situational demands. Accordingly, the presence of an attention bias (AB) for both positive and negative information may allow preferential processing of stimuli in line with the current situational goals. However, AB for negative information sometimes becomes maladaptive, being antithetical to the current adaptive needs and goals of an individual, such as in the case of affective disorders such as depression. Although difficulties in flexible shifting between emotional stimuli in depression have increasingly become a topic of discussion in the field, an integrative approach towards biased versus flexible emotional attentional processes remains absent. In the present paper, we advance a novel and integrative view of conceptualizing potentially aberrant affective attention patterns in depression as a function of the current contextual features. We propose that flexible emotional attention takes place as a result of attention prioritization towards goal-relevant emotional stimuli depending upon the current context of the individual. Specifically, the roles of context, distal and proximal goals, and approach and avoidance motivation processes are considered in a unified manner. The empirical, clinical, and interventional implications of this integrative framework provide a roadmap for future psychological and neurobiological experimental and translational research.


A Case Report of a Novel Harm Reduction Intervention Used to Detect Opioid Overdose in the Emergency Department.
As over 130 people die daily from opioid overdose in the United States, harm reduction strategies have become increasingly important. Because public restrooms are a common site for opioid overdose, emergency department waiting room restrooms (EDWRR) should be considered especially high-risk areas. We present the case of a patient found after a presumed opioid overdose in our EDWRR. Staff were alerted to his condition by a reverse motion detector (RMD), and rapidly treated him with naloxone. The RMD is a novel intervention that can save lives and should be considered in EDs with a high incidence of opioid overdose.


Atorvastatin along with imipenem attenuates acute lung injury in sepsis through decrease in inflammatory mediators and bacterial load.
Lung is one of the vital organs which is affected during the sequential development of multi-organ dysfunction in sepsis. The purpose of the present study was to examine whether combined treatment with atorvastatin and imipenem could attenuate sepsis-induced lung injury in mice. Sepsis was induced by caecal ligation and puncture. Lung injury was assessed by the presence of lung edema, increased vascular permeability, increased inflammatory cell infiltration and cytokine levels in broncho-alveolar lavage fluid (BALF). Treatment with atorvastatin along with imipenem reduced the lung bacterial load and pro-inflammatory cytokines (IL-1β and TNFα) level in BALF. The markers of pulmonary edema such as microvascular leakage and wet-dry weight ratio were also attenuated. This was further confirmed by the reduced activity of MPO and ICAM-1 mRNA expression, indicating the lesser infiltration and adhesion of inflammatory cells to the lungs. Again, expression of mRNA and protein level of iNOS in lungs was also reduced in the combined treatment group. Based on the above findings it can be concluded that, combined treatment with atorvastatin and imipenem dampened the inflammatory response and reduced the bacterial load, thus seems to have promising therapeutic potential in sepsis-induced lung injury in mice.


N-Acetyl-beta-D-glucosaminidase levels and diabetic microangiopathy.
N-Acetyl-beta-D-glucosaminidase (NAG) activity has been measured in the serum and urine of primary and secondary diabetics and in primary diabetics with microangiopathy. NAG activity has also been measured in the tears of diabetics with ocular complications and diabetics with no ocular changes. Results have shown significantly higher levels of urinary NAG in diabetics with proteinuria (p less than 0.001) and proteinuria and retinopathy (p less than 0.001). There was no correlation between urinary NAG activity and serum creatinine (r = 0.28) or urinary NAG and the degree of proteinuria (r = 0.24). Increased urinary NAG levels were also observed in secondary diabetes associated with haemochromatosis and acromegaly. Significantly higher serum NAG levels were found in newly diagnosed diabetics (p less than 0.01) and significantly lower levels in chemical diabetics (p less than 0.01). Compared to non-diabetic controls tear NAG levels were significantly higher in the diabetic controls (p less than 0.01), in diabetics with retinopathy (p less than 0.01), and in diabetics with cataract formation (p less than 0.05). An assessment of this enzyme is made in relation to the development of diabetic microangiopathy.


The proactive historian: Methodological opportunities presented by the new archives documenting genomics.
In this paper, I propose a strategy for navigating newly available archives in the study of late-twentieth century genomics. I demonstrate that the alleged 'explosion of data' characteristic of genomics-and of contemporary science in general-is not a new problem and that historians of earlier periods have dealt with information overload by relying on the 'perspective of time': the filtering effect the passage of time naturally exerts on both sources and memories. I argue that this reliance on the selective capacity of time results in inheriting archives curated by others and, consequently, poses the risk of reifying ahistorical scientific discourses. Through a preliminary examination of archives documenting early attempts at mapping and sequencing the human genome, I propose an alternative approach, in which historians proactively problematize and improve available sources. This approach provides historians with a voice in the socio-political management of scientific heritage and advances methodological innovations in the use of oral histories. It also provides a narrative framework in which to address big science initiatives by following second order administrators, rather than individual scientists. The new genomic archives thus represent an opportunity for historians to take an active role in current debates concerning 'big data' and critically embed the humanities in pressing global problems.


Non-polarisable dry electrode based on NASICON ceramic.
A NASICON-type ceramic (high sodium ion conductor) is proposed to record bioelectric signals. The electrode does not need gel before its application. The principle of the measurements is based on a sodium ion exchange between the skin and the material. Electrical measurements performed in saline solutions show that the electrode is slightly polarisable. The skin-electrode impedance was investigated. The impedance decreases as a function of the time of application. The resistive component is the major source of the impedance change. This can be explained by the perspiration process which occurs immediately with time after the application of the NASICON-based electrode on the skin. The skin condition is also an important parameter. NaCl saline solution or abrasion causes the resistance to decrease markedly.


5' Untranslated region of the Hsp12 gene contributes to efficient translation in Aspergillus oryzae.
We describe a 5' untranslated region (5'UTR) that dramatically increases the expression level of an exogenous gene in Aspergillus oryzae. Using a series of 5'UTR::GUS (uidA) fusion constructs, we analyzed the translation efficiency of chimeric mRNAs with different 5'UTRs at different temperatures. We found that the 5'UTR of a heat-shock protein gene, Hsp12, greatly enhanced the translation efficiency of the chimeric GUS mRNA at normal temperature (30 degrees C). Moreover, at high temperature (37 degrees C), the translation efficiency of the mRNA containing the Hsp12 5'UTR was far superior to that of mRNAs containing nonheat-shock 5'UTRs, resulting in much more efficient expression of GUS protein (about 20-fold higher GUS activity compared to the control construct). This 5'UTR can be used in combination with various strong promoters to enhance the expression of foreign proteins in A. oryzae.


CREB-binding protein transcription activation domain for enhanced transgene expression by a positive feedback system.
The positive feedback system using a fusion protein of the sequence-specific DNA binding domain of yeast GAL4 and the transcription activation domain of herpes simplex virus VP16 (GAL4-VP16), in which GAL4-VP16 promotes its own expression as well as that of a reporter gene product, is useful for efficient transgene expression from plasmid DNA. In this study, the transcription activation domains of endogenous proteins, instead of VP16, were fused to the GAL4 DNA binding domain, and the positive feedback systems employing the novel fusion proteins were examined. Plasmid DNAs encoding the transcription factors were introduced into mouse Hepa 1-6 cells by electroporation and lipofection. Among CREB-binding protein (226-460), sterol regulatory element-binding protein-1 (1-140), p53 (1-70), and Med15 (9-73), the CREB-binding protein functioned efficiently as an activator. These results indicated that the GAL4-CREB-binding protein is useful for enhanced transgene expression by the positive feedback system.


Polymerization contraction stress in light-cured compomer restorative materials.
The magnitude and kinetics of polymerization contraction stress build-up may be potential predictors of bond failure of adhesive restorations. The present study determined these properties of seven commercial compomers (Dyract, Dyract AP, F2000 Rasant, Hytac, Compoglass F, Luxat, Glasiosite). Polymerization shrinkage was generated by 40 s light curing the test materials (800 mW/cm2). The contraction force induced was recorded for 300 s at room temperature (23-24 degrees C) by means of a Stress-Strain-Analyzer (C factor=0.33). Maximum contraction stress (MPa), coefficient of near linear fit of contraction force/time (gradient) and relative force rate (%/s) of each material were compared with that of two hybrid composites (Tetric Ceram, Prodigy). The statistical analysis was conducted by ANOVA (alpha=0.05) and post hoc Tukey's test. No statistically significant differences in the maximum stress between Glasiosite (2.27+/-0.06 MPa), Hytac (2.31+/-0.07 MPa) and Tetric Ceram (2.21+/-0.11 MPa), and between Compoglass F (2.60+/-0.18 MPa) and Prodigy (2.70+/-0.06 MPa) were found. The contraction stress of F2000 Rasant (3.41+/-0.09 MPa) and Luxat (3.33+/-0.08 MPa) were significantly highest, whilst Dyract exhibited the significantly lowest shrinkage stress (1.27+/-0.08 MPa) among the tested materials. High contraction stress, early start of stress build-up and rapid contraction force development may lead to failure of bond to tooth structure. This study suggested that the contraction stress and kinetic behavior of compomers are generally similar to those of hybrid composites in a dry condition. Dyract might be superior in maintaining the bond with cavity walls compared to conventional hybrid composites in view of its low shrinkage stress.


Quality of life among total laryngectomized patients undergoing speech rehabilitation: correlation between several instruments.
The aim of this study was to correlate several instruments currently used for the assessment of the quality of life of patients who underwent total laryngectomy and speech rehabilitation. A cross-sectional, observational study was conducted with 38 patients after total laryngectomy and speech therapy aiming to develop oesophageal speech. The patients were divided into the following two groups (19 participants each): speakers and non-speakers. The quality of life instruments used were as follows: visual analogue scale (VAS); Voice Handicap Index (VHI); Voice-Related Quality of Life (V-RQOL); Functional Assessment of Cancer Therapy - Head & Neck (FACT-H&N); European Organisation for Research and Treatment of Cancer Core Quality of Life Questionnaire (EORTC QLQ-C30); European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-Head and Neck (EORTC QLQ-H&N35); and University of Washington Quality of Life (UW-QOL). The V-RQOL global health domain exhibited a strong correlation with the VHI. The EORTC QLQ-C30 exhibited a moderate to strong correlation with the EORTC QLQ-H&N35 functional domain in both groups. The EORTC QLQ-C30 functional domain exhibited a strong to moderate correlation with all other instruments in both groups. The UW-QOL exhibited a moderate to strong correlation with the VHI and EORTC QLQ-C30 in both groups. The EORTC QLQ-C30, EORTC QLQ-H&N35 and UW-QOL were the instruments that most correlated with the remaining instruments, indicating that any of the three can be used to assess the quality of life of the target population regardless of oesophageal voice development.


A new microphallid (Digenea) species from Lontra provocax (Mammalia: Mustelidae) from freshwater environments of northwestern Patagonia (Argentina).
A new microphallid species of Maritrema is described from the native southern river otter, Lontra provocax (Thomas). A naturally infected otter was found dead in the Nahuel Huapi National Park, Argentina. Ovigerous adult worms were recovered from the anterior portion of the intestine. Specimens of Maritrema huillini n. sp. have an unarmed genital pore and glabrous cirrus. They can be distinguished from all other species in the genus by having a long intestinal ceca extending up to three-quarters of the testes length to the level of the posterior border of the testes and a metraterm composed of a proximal sphincter, a non-muscular sac, and a distal muscular portion. This microphallid is the first species recovered from a South American eutherian host and the first digenean recorded for L. provocax.


Systematic Bayesian posterior analysis guided by Kullback-Leibler divergence facilitates hypothesis formation.
Bayesian inference produces a posterior distribution for the parameters of a mathematical model that can be used to guide the formation of hypotheses; specifically, the posterior may be searched for evidence of alternative model hypotheses, which serves as a starting point for hypothesis formation and model refinement. Previous approaches to search for this evidence are largely qualitative and unsystematic; further, demonstrations of these approaches typically stop at hypothesis formation, leaving the questions they raise unanswered. Here, we introduce a Kullback-Leibler (KL) divergence-based ranking to expedite Bayesian hypothesis formation and investigate the hypotheses it generates, ultimately generating novel, biologically significant insights. Our approach uses KL divergence to rank parameters by how much information they gain from experimental data. Subsequently, rather than searching all model parameters at random, we use this ranking to prioritize examining the posteriors of the parameters that gained the most information from the data for evidence of alternative model hypotheses. We test our approach with two examples, which showcase the ability of our approach to systematically uncover different types of alternative hypothesis evidence. First, we test our KL divergence ranking on an established example of Bayesian hypothesis formation. Our top-ranked parameter matches the one previously identified to produce alternative hypotheses. In the second example, we apply our ranking in a novel study of a computational model of prolactin-induced JAK2-STAT5 signaling, a pathway that mediates beta cell proliferation. Within the top 3 ranked parameters (out of 33), we find a bimodal posterior revealing two possible ranges for the prolactin receptor degradation rate. We go on to refine the model, incorporating new data and determining which degradation rate is most plausible. Overall, while the effectiveness of our approach depends on having a properly formulated prior and on the form of the posterior distribution, we demonstrate that our approach offers a novel and generalizable quantitative framework for Bayesian hypothesis formation and use it to produce a novel, biologically-significant insight into beta cell signaling.


REtrieval And cure of Chronic Hepatitis C (REACH): Results of micro-elimination in the Utrecht province.
The Netherlands is one of the six European countries considered on track to eliminate hepatitis C virus by 2030. To achieve this goal, continuous efforts have to be put into designing efficient case-finding strategies, including the retrieval of previously diagnosed hepatitis C virus-infected who are lost to follow-up. To trace and treat all lost to follow-up hepatitis C virus patients in the Utrecht region and create an efficient retrieval strategy that can be used in future (national) retrieval initiatives. Positive hepatitis C virus diagnostic tests (anti-hepatitis C virus IgG or hepatitis C virus-RNA) from the laboratory of all four hospitals and one central laboratory for primary care diagnostics in the province of Utrecht from 2001 to 2015 were linked to clinical records. Untreated patients with available contact information were deemed eligible for retrieval and invited for reevaluation with (virology) blood tests, fibroscan measurement and possible direct-acting antiviral therapy. After screening all hepatitis C virus diagnostics, 1913 chronic hepatitis C virus-infected were identified of which 14.1% (n = 269) were invited back into care. Overall, 17.4% was traced with the highest yield (28.3%) in those who lived in the Utrecht province. Through renewed patient assessments, 42 chronic hepatitis C virus infections were re-identified (76% with a history of intravenous drug use, 24% with Metavir F3-F4). Until now, 59% has either scheduled or initiated direct-acting antiviral therapy. The retrieval of previously diagnosed hepatitis C virus patients through screening of laboratory diagnostics from the past is feasible and should be pursued for further control and reduction of hepatitis C virus infection. Retrieval is most successful when performed regionally. To completely eliminate chronic hepatitis C virus (HCV) infection and prevent complications, undiagnosed and also previously diagnosed but lost to follow-up (LFU) HCV patients have to be brought (back) into care for therapy. Retrieval of LFU HCV patients through screening of laboratory diagnostics from the past is feasible and most successful when performed regionally.


Effect of thyroid state on cytosolic free calcium in resting and electrically stimulated cardiac myocytes.
The effects of the thyroid state on the cytosolic free Ca2+ concentration, [Ca2+]i, of resting and K+-depolarized cardiomyocytes were studied using the fluorescent Ca2+ indicator fura2. The mean resting [Ca2+]i in euthyroid myocytes (89 +/- 8 nM) was not significantly different from that in hyperthyroid myocytes (100 +/- 14 nM). The resting O2-consumption rate was identical for both groups when expressed per mg protein, but a 35% higher value was observed in the hyperthyroid group when expressed per cell on account of the cellular hypertrophy induced by thyroid hormone. Potassium induced depolarization (50 mM [K+]0) raised the level of [Ca2+]i by 50% in both groups. When ATP-coupled respiration was blocked with oligomycin, the 50 mM K+-induced rise in [Ca2+]i was accompanied in both groups by a 40% rise in glycolytic activity as inferred from measurement of lactate production. Ca2+-fluorescence transients were recorded from electrically stimulated myocytes of euthyroid, hyperthyroid and hypothyroid rats. The time taken to reach peak fluorescence (TPL) and that to 50% decay of peak fluorescence (RL0.5) decreased in the direction hypothyroid----hyperthyroid, indicating an increase in Ca2+ fluxes in the same direction. Isoproterenol (1 microM) enhanced the peak Ca2+ fluorescence in electrically stimulated hypothyroid and euthyroid myocytes but not in hyperthyroid myocytes. Both the TPL and RL0.5 were decreased by isoproterenol in euthyroid, but more so in hypothyroid myocytes. None of these parameters were influenced by isoproterenol in the hyperthyroid group. We conclude that (1) thyroid hormone increases neither the O2-consumption rate nor the level of [Ca2+]i of resting cardiomyocytes and (2) the effects of the beta-receptor-agonist isoproterenol on Ca2+ transients of electrically stimulated myocytes, are inversely related to the documented changes in beta-receptor density in heart tissue occurring with alterations in the thyroid state.


Vaccination against Staphylococcus aureus mastitis in two Swedish dairy herds.
Staphylococcus aureus is a common udder pathogen in dairy cows, and may cause severe mastitis problems in some herds. In herds where normal control measures are not successful, vaccination might be an additional tool to use if sufficiently efficient. The aim of the present study was to evaluate the efficacy of a commercially available vaccine (Startvac(®), Hipra, Spain) in two commercial Swedish dairy herds where the control programs for S. aureus mastitis had been unsuccessful. Within each herd cows were randomly assigned to vaccine or control groups, and effects on udder health and milk production during 120 days after calving, and survival during the following lactation were evaluated. A field study was performed in two high producing Swedish herds having approximately 600 (herd A) and 200 (herd B) cows. During 12 months, cows with odd numbers were vaccinated three times around calving according to label protocol, while cows with even numbers constituted the not vaccinated control group. Quarter milk samples for bacteriological culturing were collected from all cases of clinical and subclinical mastitis. The outcome was evaluated during 120 days after calving using data on SCC and daily milk yield at monthly milk recordings, and incidence of mastitis due to S. aureus, coagulase-negative staphylococci, streptococci and coliforms. Cow survival throughout lactation was also studied. In herd A, 239 and 240 cows were included in the vaccinated and control groups, respectively. Corresponding numbers for herd B was 126 and 151 cows. Significant differences between vaccinated and control groups were not found in any of the parameters investigated. Vaccination with a commercial polyvalent vaccine did not have any beneficial effects on udder health, milk production or survival in two commercial dairy herds with mastitis problems due to S. aureus.


Salt has contrasting effects on the digestive processing of dilute nectar by two Neotropical nectarivorous bats.
Nectarivorous vertebrates might include sugar-dilute nectar in their diet and they are expected to undergo compensatory feeding. However, physiological constraints might limit the intake of sugar-dilute nectar, affecting energy budgets. Among other physiological processes, the limiting role of osmoregulation is supported by enhanced intake rate of dilute sugar solutions by avian nectarivores when salt is added. We tested if the Greater Antillean Long-tongued bat (Monophyllus redmani) and the Brown flower bat (Erophylla sezekorni) compensated energy intake when fed dilute-sugar solutions (2.5 and 5% sucrose), and if salt content (11, 20 and 40 mM NaCl l-1) modulated the intake rate of these solutions. Both species were unable to compensate intake of solutions with varying sugar densities, and energy intake on the 2.5 and 5% diets was lower than on the most concentrated diets (10, 20 and 30% sucrose). Both species responded differently to the addition of salt. Salt addition did not affect the intake of 2.5% sugar solutions by the Greater Antillean Long-tongued bat, and it decreased the intake of 5% sugar solutions. In contrast, the Brown flower bat increased the intake of 2.5 and 5% sugar solutions when salt was added. Intake responses to varying sugar densities of our two focal species and that of other bat species previously studied indicate that they are not uniform and that they might be modulated by digestive and osmoregulatory physiological traits.


Effects of copper chloride on formation of polychlorinated dibenzofurans in model waste incineration in a laboratory-scale fluidized-bed reactor.
Combustion experiments in a laboratory-scale fluidized-bed reactor have been performed to clarify the effects of copper chloride as a catalyst on polychlorinated dibenzofurans (PCDFs) formation in municipal waste incineration. We used model wastes with and without copper chloride (CuCl2 x 2H2O) as a catalyst, both of which contained polyvinyl chloride (PVC) as a chlorine source. Combustion temperature was set to 900 degrees C, and the amount of air supplied was twice as much as the theoretical amount. The experimental setup had been carefully planned to ensure avoidance of the influences of previous experiments. Results of these present experiments revealed that copper chloride in the waste increased the amount of PCDFs formed and made the homologue profile shifted towards the highly chlorinated species. Copper chloride contributes to PCDFs formation by promoting chlorination via catalytic reactions, whereby the reaction could be important in that organic matters are chlorinated directly by chlorinated compounds related to Deacon reaction such as copper chloride. It was elucidated that characteristic isomer distribution patterns appeared in case the waste contained copper chloride. It is probable in our experiment with copper chloride that PCDFs are mainly formed via catalytic reactions of copper compounds and carbon.


Pancreatic epithelial cells can be converted into insulin-producing cells by GLP-1 in conjunction with virus-mediated gene transfer of pdx-1.
Glucagonlike peptide-1 (GLP-1) stimulates insulin secretion and proliferation by islet cells in vitro and in vivo, associated with an activation of pancreatic duodenal homeobox gene-1 (pdx-1) function. The effect of GLP-1 on the conditionally immortalized pancreatic epithelial cells (IMPE cells) is not clear when they are treated in conjunction with the adenovirus-mediated gene transfer of pdx-1. IMPE cells were established from the pancreas of H-2K(b)-tsA58 transgenic mice. IMPE cells were maintained at 33 degrees C with 10 U/mL interferon (IFN)-gamma and the experiments were performed at 39 degrees C without IFN-gamma. IMPE cells were infected with 20 multiplicities of Ad-pdx-1 or control Ad-lacZ at 39 degrees C without IFN-gamma and were incubated with various concentrations of GLP-1. After 48 hours, immunofluorescence and reverse transcriptase-polymerase chain reaction for insulin and pdx-1 expression were examined. Immunoreactive insulin in the cell lysate and supernatant was also analyzed. The glucose concentration in the culture medium was changed to test the insulin secretory responsiveness of the IMPE cells. The treatment with GLP-1 in conjunction with Ad-pdx-1 induced insulin production by IMPE cells, but the treatment with either GLP-1 or Ad-pdx-1 alone failed to induce insulin production. Insulin production and secretion were increased by GLP-1 and by glucose in a dose-dependent manner. In addition, the insulin-producing IMPE cells acquired a rapid insulin secretory responsiveness to the changes of extracellular glucose concentration. GLP-1 and pdx-1 work together to induce insulin-producing cells from IMPE cells, which bear unique characteristics of pancreatic ductal cells. The results suggest that GLP-1 may be another important determiner of pancreatic endocrine differentiation as is pdx-1.


Construction of Escherichia coli strains producing L-serine from glucose.
L-Serine is usually produced from glycine. We have genetically engineered Escherichia coli to produce L-serine from glucose intracellularly. D-3-Phosphoglycerate dehydrogenase (PGDH, EC 220.127.116.11) in E. coli catalyzes the first committed step in L-serine formation but is inhibited by L-serine. To overcome this feedback inhibition, both the His(344) and Asn(346) residues of PGDH were converted to alanine and the mutated PGDH (PGDH(dr)) became insensitive to L-serine. However, overexpression of PGDH(dr) gave no significant increase of L-serine accumulation but, when L-serine deaminase genes (sdaA, sdaB and tdcG) were deleted, serine accumulated: (1) deletion of sdaA gave up to 0.03 mmol L-serine/g; (2) deletion of both sdaA and sdaB accumulated L-serine up to 0.09 mmol/g; and (3) deletion of sdaA, sdaB and tdcG gave up to 0.13 mmol L-serine/g cell dry wt.


Sonochemical in situ immobilization of Pd nanoparticles on green tea extract coated Fe3O4 nanoparticles: An efficient and magnetically recyclable nanocatalyst for synthesis of biphenyl compounds under ultrasound irradiations.
This work describes (i) an eco-friendly approach for in situ immobilization of Pd nanoparticles on the surface of Fe3O4 nanoparticles, with help of green tea extract and ultrasound irradiations, without using any toxic reducing agents and (ii) development of ultrasound assisted simple protocol for synthesis of biphenyl compounds. The structural, morphological and physicochemical characteristics of the catalyst were determined by different analytical methods including inductively coupled plasma (ICP) analysis, Fourier transform infrared spectroscopy (FT-IR), field emission scanning electron microscopy (FESEM), energy dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) and vibrating sample magnetometry (VSM). Catalytic performance of Pd/Fe3O4 NPs as magnetic and heterogeneous catalyst was evaluated in synthesis of various biphenyl compounds throughout Suzuki coupling reactions by using the ultrasound-assisted method that was developed in this study. The Pd/Fe3O4 NPs demonstrated a noticeable catalytic activity by giving high product yields. Furthermore, the heterogeneous nanocatalyst was successfully recovered up to six times without significant activity loss. Additionally, with respect to conventional coupling reactions, the ultrasound-assisted synthesis reactions presented the advantages of green conditions, short reaction times, high yields and easier work-up.


Microglial phagocytosis is enhanced by monomeric alpha-synuclein, not aggregated alpha-synuclein: implications for Parkinson's disease.
Gathering evidence has associated activation of microglia with the pathogenesis of numerous neurodegenerative diseases of the central nervous system (CNS) such as Alzheimer's disease and Parkinson's disease. Microglia are the resident macrophages of the CNS whose functions include chemotaxis, phagocytosis, and secretion of a variety of cytokines and proteases. In this study, we examined the possibility that alpha-synuclein (alpha-syn), which is associated with the pathogenesis of Parkinson's disease, may affect the phagocytic function of microglia. We found that extracellular monomeric alpha-syn enhanced microglial phagocytosis in both a dose- and time-dependent manner, but beta- and gamma- syn did not. We also found that the N-terminal and NAC region of alpha-syn, especially the NAC region, might be responsible for the effect of alpha-syn on microglial phagocytosis. In contrast to monomeric alpha-syn, aggregated alpha-syn actually inhibited microglial phagocytosis. The different effects of monomeric and aggregated alpha-syn on phagocytosis might be related to their localization in cells. This study indicates that alpha-syn can modulate the function of microglia and influence inflammatory changes such as those seen in neurodegenerative disorders.


The impact of C-reactive protein-albumin-lymphocyte (CALLY) index on the prognosis of patients with distal cholangiocarcinoma following pancreaticoduodenectomy.
The C-reactive protein (CRP)-albumin-lymphocyte (CALLY) index is a novel inflammation-based biomarker, which has been associated with long-term outcomes in patients with hepatocellular carcinoma. We aimed to investigate whether the CALLY index can predict the prognosis for distal cholangiocarcinoma after pancreaticoduodenectomy. The study comprised 143 patients who had undergone primary pancreaticoduodenectomy for distal cholangiocarcinoma between 2002 to 2019. The CALLY index was defined as (albumin × lymphocyte)/ (CRP × 104). We investigated the association of CALLY index with disease-free survival and overall survival by univariate and multivariate analyses. Eighty-seven (61%) patients had a preoperative CALLY index <3.5. In multivariate analysis, obstructive jaundice drainage (P < .01), poorly differentiated tumor (P < .01), and CALLY index<3.5 (P = .02) were independent predictors of disease-free survival, while obstructive jaundice drainage (P < .01), poorly differentiated tumor (P < .01), and CALLY index <3.5 (P = .02) were independent predictors of overall survival. The CALLY index may be an independent and significant indicator of poor long-term outcomes in patients with distal cholangiocarcinoma after pancreaticoduodenectomy, suggesting the importance of comprehensive assessment for inflammatory status.


Novel domain-specific POU3F4 mutations are associated with X-linked deafness: examples from different populations.
Mutations in the POU3F4 gene cause X-linked deafness type 3 (DFN3), which is characterized by inner ear anomalies. Three Turkish, one Ecuadorian, and one Nigerian families were included based on either inner ear anomalies detected in probands or X-linked family histories. Exome sequencing and/or Sanger sequencing were performed in order to identify the causative DNA variants in these families. Four novel, c.707A>C (p.(Glu236Ala)), c.772delG (p.(Glu258ArgfsX30)), c.902C>T (p.(Pro301Leu)), c.987T>C (p.(Ile308Thr)), and one previously reported mutation c.346delG (p.(Ala116ProfsX26)) in POU3F4, were identified. All mutations identified are predicted to affect the POU-specific or POU homeo domains of the protein and co-segregated with deafness in all families. Expanding the spectrum of POU3F4 mutations in different populations along with their associated phenotypes provides better understanding of their clinical importance and will be helpful in clinical evaluation and counseling of the affected individuals.


Secular Trends in Preeclampsia Incidence and Outcomes in a Large Canada Database: A Longitudinal Study Over 24 Years.
The incidence of preeclampsia is increasing, but effects on women and infants are unclear. We measured the incidence of preeclampsia over time in a large Canadian population, and assessed trends in maternal and infant morbidity and mortality. We carried out a population-based study of 1,901,376 linked hospital discharge abstracts for all deliveries in the province of Quebec, Canada from 1989 through 2012. We computed the annual incidence of preeclampsia, and used log binomial models to determine associations with severe morbidity and mortality for preeclamptic vs nonpreeclamptic pregnancies. Main outcomes included maternal, fetal, and infant mortality, admission to intensive care, intubation, preterm delivery, growth restriction, cesarean delivery, transfusion, and severe medical complications. The incidence of preeclampsia increased from 26.4 per 1000 deliveries in 1989 to 50.6 in 2012. Maternal, fetal, and infant mortality decreased with time for preeclamptic but not for nonpreeclamptic pregnancies. By 2007-2012, risk for women with preeclampsia had declined for most maternal morbidities, except acute renal failure, which increased relative to no preeclampsia (risk ratio, 21.5; 95% confidence interval, 16.9-27.3). Risk of infant morbidity also decreased, but this coincided with an increase in the excess number of intubations and admissions for intensive care for preeclampsia relative to no preeclampsia. The incidence of preeclampsia increased during the study, but with little effect on the risk of maternal and infant morbidity and mortality. For most outcomes, the risk decreased relative to no preeclampsia, with more aggressive medical management over time.


Frequency of early and late-onset dementias in a Japanese memory disorders clinic.
The aim of this study was to compare the diagnostic profiles of patients with early (age<65 years) and late (age>or=65 years) onset of dementia in a memory disorders clinic in Japan. A total of 512 consecutive memory clinic patients were evaluated using clinical information and results of examinations. Diagnosis of dementia was made according to DSM-III-R, and that of subtypes according to standard diagnostic criteria. A total of 464 patients met the criteria for dementia. Amongst late-onset patients (n=430), Alzheimer's disease (AD) (48.1%) was the most frequent cause of dementia, followed by AD with cerebrovascular disease (CVD) (31.4%), vascular dementia (VaD) (9.1%), dementia with Lewy bodies (DLB) (3.7%), frontotemporal lobar degeneration (FTLD) (1.6%), and others (5.8%). On the contrary, amongst early onset patients (n=34), the most common dementia diagnosis was AD (38.2%), followed by VaD (23.5%), FTLD (14.7%), AD with CVD (5.9%), DLB (2.9%), and others (17.6%). FTLD and VaD were significantly more common in the early onset group. All patients, but one, with DLB and Parkinson's disease dementia were late-onset. The relative frequencies of AD, VaD, and DLB in our series are consistent with epidemiologic findings in several Western countries; however, the frequency of FTLD is not consistent with the previous findings presenting high frequency in late-onset patients in some Western countries.


Vascular morphology and expression of angiogenic factors in sheep ovaries and their relation with the lambing rate.
The study aims to present the vascular anatomy of ovarian arterial system in sheep and to explore its relation with the angiogenic factors vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2), environmental factors and the lambing rate. Small-tailed Han sheep, Hu sheep, Aohan finewool sheep and Tibetan sheep who are from different habitats in China and with different lambing rates were investigated. The vascular corrosion casting technique and scanning electron microscopy were used to understand arterial architecture, and real-time quantitative PCR (qRT-PCR) and Western blotting were used for assessing VEGF and Ang-2 expression. Results showed that high-prolificity breeds had significantly larger vessel diameters in ovaries than low-prolificity ones. VEGF gene transcript and protein expression were related with the lambing rate, while Ang-2 gene transcript was negatively related with the lambing rate. Results indicated that high-prolificity breeds have more abundant arterial blood supply, which is associated with VEGF expression.


Phases of the dentition for the assessment of skeletal maturity: a diagnostic performance study.
The aim of this study was to analyze the relationship between the circumpubertal phases of the dentition (early mixed, intermediate mixed, late mixed, early permanent) and skeletal maturity as assessed by means of the cervical vertebral maturation (CVM) method. The sample of 1000 subjects included 250 (125 boys, 125 girls) in each of the 4 dentition phases. Individual skeletal maturity was determined by using the CVM method. The relationship between the skeletal maturity (stages in CVM) and the phases of the dentition was evaluated statistically by means of indicators of diagnostic test performance. Prepubertal stage 1 (CS1) was the variable diagnosed in the early mixed and intermediate mixed dentitions; pubertal stage 3 (CS3) was the variable diagnosed in the late mixed and early permanent dentitions. The early mixed dentition phase showed a strong diagnostic value for the identification of prepubertal skeletal maturity (CS1), whereas the intermediate mixed dentition phase had a low diagnostic value for the same prepubertal stage. Neither the late mixed dentition nor the early permanent dentition appeared to be a valid indicator for the onset of the pubertal growth spurt.


Identification of candidate biomarkers and pathways associated with type 1 diabetes mellitus using bioinformatics analysis.
Type 1 diabetes mellitus (T1DM) is a metabolic disorder for which the underlying molecular mechanisms remain largely unclear. This investigation aimed to elucidate essential candidate genes and pathways in T1DM by integrated bioinformatics analysis. In this study, differentially expressed genes (DEGs) were analyzed using DESeq2 of R package from GSE162689 of the Gene Expression Omnibus (GEO). Gene ontology (GO) enrichment analysis, REACTOME pathway enrichment analysis, and construction and analysis of protein-protein interaction (PPI) network, modules, miRNA-hub gene regulatory network and TF-hub gene regulatory network, and validation of hub genes were performed. A total of 952 DEGs (477 up regulated and 475 down regulated genes) were identified in T1DM. GO and REACTOME enrichment result results showed that DEGs mainly enriched in multicellular organism development, detection of stimulus, diseases of signal transduction by growth factor receptors and second messengers, and olfactory signaling pathway. The top hub genes such as MYC, EGFR, LNX1, YBX1, HSP90AA1, ESR1, FN1, TK1, ANLN and SMAD9 were screened out as the critical genes among the DEGs from the PPI network, modules, miRNA-hub gene regulatory network and TF-hub gene regulatory network. Receiver operating characteristic curve (ROC) analysis confirmed that these genes were significantly associated with T1DM. In conclusion, the identified DEGs, particularly the hub genes, strengthen the understanding of the advancement and progression of T1DM, and certain genes might be used as candidate target molecules to diagnose, monitor and treat T1DM.


Blunted neural response to emotional faces in the fusiform and superior temporal gyrus may be marker of emotion recognition deficits in pediatric epilepsy.
Individuals with epilepsy are at risk for social cognition deficits, including impairments in the ability to recognize nonverbal cues of emotion (i.e., emotion recognition [ER] skills). Such deficits are particularly pronounced in adult patients with childhood-onset seizures and are already evident in children and adolescents with epilepsy. Though these impairments have been linked to blunted neural response to emotional information in faces in adult patients, little is known about the neural correlates of ER deficits in youth with epilepsy. The current study compared ER accuracy and neural response to emotional faces during functional magnetic resonance imaging (fMRI) in youth with intractable focal epilepsy and typically developing youth. Relative to typically developing participants, individuals with epilepsy showed a) reduced accuracy in the ER task and b) blunted response to emotional faces (vs. neutral faces) in the bilateral fusiform gyri and right superior temporal gyrus (STG). Activation in these regions was correlated with performance, suggesting that aberrant response within these face-responsive regions may play a functional role in ER impairments. Reduced engagement of neural circuits relevant to processing socioemotional cues may be markers of risk for social cognitive deficits in youth with focal epilepsy.


Diverse recruitment strategies result in different participation percentages in a web-based study, but in similar compliance.
We compared participation and compliance with a web-based data collection on infections among population-based samples recruited in different ways. Individuals were recruited from participants in the German National Cohort study (Group A, n = 279) or persons who were invited to this study but did not participate (Group B, n = 53). A third group was invited to the web-based study only (Group C, n = 145). Response varied among groups between 3 % (B), 11 % (C) and 61 % (A), but compliance was similar (81-85 %). Response did not differ by age and sex. Compliance was lower among the youngest and oldest participants. In addition, participants currently not employed were more likely to have better compliance. Semi-parametric group-based modelling identified three distinct compliance trajectories; "poor compliance" (8 %), "improving compliance" (14 %) and "very good compliance" (78 %). Participation differed among modes of recruitment, but compliance was similar among groups and notably high. Different recruitment approaches can be used and collected data can be combined to achieve greater sample sizes for longitudinal web-based studies.


Drug usage analysis in pregnant women.
Drug usage in pregnant women is associated with a problem of possible negative influence on prenatal development. It is always necessary to judge the need for drug administration during pregnancy. The aim of presented study was to analyse data about pregnant women hospitalized in the postpartum period. The study was designed as a retrospective observational study including 300 women hospitalized at the 2nd Department of Gynaecology and Obstetrics, University Hospital, Bratislava. Data were obtained through questionnaires in form of an interview. The average age of women was 30.79 ± 4.40 years. Risk pregnancy occured in 20.59 % of women. Chronic disorders before pregnancy required regular pharmacotherapy in 29.24 %. Drug usage analysis: I. trimester, 31 % used at least one drug, 52 % nutritional supplements, 63.3 % drug and/or nutritional supplement; II. trimester, 23 % used at least one drug, 45 % nutritional supplements, 58.3 % drug and/or nutritional supplement; III. trimester, 32 % used at least one drug, 67 % nutritional supplements, 75.3 % drug and/or nutritional supplement. Drug usage during pregnancy requires great precaution at choosing pharmacotherapy. The benefit of pharmacotherapy should always outweight the potential risk of administered drug (Tab. 3, Fig. 3, Ref. 37).


The blood composition of different breeds of bulls undergoing beef performance tests.
Blood samples taken on three occasions from each of 66 bulls undergoing beef performance tests were analysed for packed cell volume, blood glucose, haemoglobin, serum albumin, urea nitrogen, total protein, inorganic phosphate, Ca, Ca, Mg, K, Na, Cu, Fe and total iron binding capacity. The bulls were individually fed and situated at two centres, one accommodating Lincoln-Red, Devon and Sussex breeds and the other, Hereford. Significant differences between the Lincoln-Red, Devon and Sussex breeds were observed in concentrations of glucose, urea nitrogen, total iron binding capacity (P less than 0.001), Ca (P less than 0.01), Na and Cu (P less than 0.05). There were no significant correlations between the growth rates of individual bulls and their blood composition.


Possible nosocomial transmission of group Y Neisseria meningitidis among oncology patients.
Within 4 days, two oncology patients in the Clinical Research Unit had bacteremia causes by group Y Neisseria meningitidis. Three additional patients, identified by prevalence survey, were found to have nasopharyngeal colonization with that serogroup, compared with only one employee (not associated with patients) and no family contacts. Infected and colonized (case) patients were located in the same or adjacent rooms but did not have close contact. A comparison of host and risk factors showed no significant differences between case patients and the other (control) patients located in the same rooms. In retrospect, the index patient, who had marked sputum production but was not isolated, had unrecognized meningococcal pneumonia and probably was the hospital source of a heavy, airborne dispersal of organisms to other patients. Respiratory isolation is warranted for patients with suspected N. meningitidis infection.


Newborns and preterm infants at term equivalent age: A semi-quantitative assessment of cerebral maturity.
Currently available MRI scoring systems of cerebral maturation in term and preterm infant at term equivalent age do not include the changes of transient fetal compartments that persist to term age. We studied the visibility and the pattern of these structures in healthy term newborns compared to preterm infants at term equivalent age in order to investigate if they can be included in a new MRI score system. We hypothesized that transient fetal compartments are different in both groups, and that these differences can be characterized using the clinical T2-weighted MRIs. Using 3T MRI T2-weighted brain sequences of 21 full-term and 41 preterm infants (< 32 weeks), scanned at term equivalent age, 3 raters independently scored the maturation level of 3 transient fetal compartments: the periventricular crossroads, von Monakow segments of the white matter, and the subplate compartment. These 3 new items were included in a scoring system along with validated parameters of brain maturation (germinal matrix, bands of migration, subarachnoid space and quality of gyrification). A cumulative maturity score was calculated separately for both groups of newborns by adding together each item. More mature were the brain structures, higher was the cumulative maturity score. Cumulative maturity score distinguished full-term from preterm infants (mean score 41/60 ± 1.4 versus 37/60 ± 2.5 points, p < 0.001), with an increase of 0.5 points for each supplemental gestational week at birth (r = 0.5, 95% CI 0.5 - 0.85). While a majority of transient fetal compartments were less mature in preterm group at term equivalent age, von Monakow segments of the white matter and subplate compartment presented a more advanced maturational stage in the preterm group compared to the term group. No subject had all scored items in the most mature state. Except a slight intra-rater agreement for von Monakow segment II, inter- and intra-rater agreements were moderate to excellent indicating the potential of the developed scoring system in routine clinical practice. Brain transient fetal structures can be assessed on regular T2-weighted MRI in newborns. Their appearance differs between term and preterm babies. However our results suggest a more complex situation, with both delayed and accelerated maturation pattern in preterm infants. It remains to be determined if these differences could be biomarkers of the future neurodevelopment of preterm infants.


Regulatory T cells: potential target in anticancer immunotherapy.
The concept of regulatory T cells was first described in the early 1970s, and regulatory T cells were called suppressive T cells at that time. Studies that followed have demonstrated that these suppressive T cells negatively regulated tumor immunity and contributed to tumor growth in mice. Despite the importance of these studies, there was extensive skepticism about the existence of these cells, and the concept of suppressive T cells left the center stage of immunologic research for decades. Interleukin-2 receptor alpha-chain, CD25, was first demonstrated in 1995 to serve as a phenotypic marker for CD4+ regulatory cells. Henceforth, research of regulatory T cells boomed. Regulatory T cells are involved in the pathogenesis of cancer, autoimmune disease, transplantation immunology, and immune tolerance in pregnancy. Recent evidence has demonstrated that regulatory T cell-mediated immunosuppression is one of the crucial tumor immune evasion mechanisms and the main obstacle of successful cancer immunotherapy. The mechanism and the potential clinical application of regulatory T cells in cancer immunotherapy are discussed.


Effect of serum lipoproteins of bile obstructed rats on 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in perfused rat liver.
Total lipoproteins as well as fractionated VLDL + LDL and HDL from fasted control rats and bile-ligated rats were tested in liver perfusion for their effect on 3-hydroxy-3-methylglutaryl CoA reductase activity in normal rat livers. The total lipoproteins of bile-obstructed rats had 3 times greater capacity to increase 3-hydroxy-3-methylglutaryl CoA reductase activity than that of the control total lipoproteins. When the fractionated lipoproteins were tested from fasted control rats, it was found that the major stimulating activity was in the HDL fraction with minor activity in the VLDL + LDL fraction. When these plasma components isolated from fasted bile-ligated rats were tested, it was found that the major activity had shifted to the VLDL + LDL fraction with the HDL having only a minor stimulatory role. The possible mechanism of action of the abnormal lipoproteins associated with bile obstruction in regulating 3-hydroxy-3-methylglutaryl CoA reductase activity is discussed.


[Nucleic acid and protein content in rat prostate during combined treatment with the gonadorelin agonist and an androgen receptor inhibitor].
The effects of androgen receptor blocker Flutamide (Niftolide) and/or LH-releasing hormone agonist Surfagon on the nucleic acids and protein contents in the ventral prostate of male rats were studied. It was shown that combined drug treatment led to potentiating of "antiprostatic" effects of both preparations resulted in significant decrease of DNA, RNA and protein contents.


Interoception: Probing internal state is inherent to perception and cognition.
Interoception, i.e. the perception of the organism physiological state, jointly operates with perception and cognition, including when homeostatic balance is not particularly challenged. Studying interoception offers new leads to understanding the link between internal state, neural activity, and behavior.


The relation of urinary estrogen metabolites with mammographic densities in premenopausal women.
Mammographic density is a strong predictor of breast cancer risk. The total amount and the metabolism of endogenous estrogens, e.g., the ratio of 2-hydroxyestrone (2-OHE(1)) and 16α-OHE(1) may influence breast cancer risk. This study examined the association of urinary estrogen metabolites with breast density in premenopausal women. Urine samples were collected at baseline and after 2 years, analyzed for 11 estrogen metabolites plus progesterone and testosterone by liquid chromatography mass spectrometry, and adjusted for creatinine levels. Mixed-effects regression was applied to examine the association of estrogens with breast density. Total estrogen metabolites (181 ± 113 vs. 247 ± 165 pmol/mg creatinine, p=0.01) and the 2/16α-OH ratio (8.4 ± 10.4 vs. 13.0 ± 17.1, p=0.02) were lower in the 74 Asian than in the 114 non-Asian women. In adjusted models, positive associations of total estrogen metabolites (p=0.002) and the 2/16α-OHE(1) ratio (p=0.08) with percent density were detected in Asians only. In all women, mammographic density was positively associated with the 2-OH pathway (p=0.01), inversely related to the 16α-OH pathway (p=0.01), and not associated with the 4-OH pathway, testosterone, and progesterone. Results for the size of the dense area weakly reflected the findings for percent density, while associations with the non-dense area were in the opposite direction. The findings that the 2-OH pathway is associated with higher and the 16α-OH pathway with lower breast density contradicts the hypothesized risk profile of these metabolites, but, if a relation between estrogen metabolites and breast cancer risk exists, it may be mediated through pathways other than mammographic density.


[Use of charcoal enterosorbents in the complex treatment of patients with obstructive jaundice. Report II. Experimental study].
In the experiments on rats with modelled bile duct obstruction and hepatic failure, the pronounced detoxicative value of the SKN P-1 and SKN P-2 carbonic sorbents is proved.


Long-term exposure to extremely low-frequency magnetic fields impairs spatial recognition memory in mice.
1. In the present study, we investigated the short- and long-term effects of extremely low-frequency (ELF) magnetic fields on spatial recognition memory in mice by using a two-trial recognition Y-maze that is based on the innate tendency of rodents to explore novel environments. 2. Mice were exposed to 25 or 50 Hz electromagnetic fields for either 7 (short term) or 25 days (long term) and then tested in the Y-maze. 3. The results indicated that neither short- nor long-term exposure to magnetic fields affected the locomotor activity of mice in the Y-maze. However, long-term exposure to 50 Hz fields reduced recognition of the novel arm. 4. Our findings suggest that ELF magnetic fields impair spatial recognition memory in the Y-maze depending on the field strength and/or duration of exposure.


Paraneoplastic antibodies against HuD detected by a sensitive radiobinding assay.
Patients with paraneoplastic encephalomyelitis(subacute sensory neuronopathy) (PEM/SSN), most commonly associated with small-cell lung cancer (SCLC), frequently harbor Hu antibodies, which are usually detected by indirect immunohistochemistry or immunoblot. We developed a new radioimmunobinding assay to detect Hu antibodies based on in vitro transcribed and translated (ITT) HuD. High levels of Hu antibodies were detected in all seven PEM/SSN patients tested, but not in any of 15 patients with other paraneoplastic syndromes, 20 patients with Sjögren's syndrome, 20 patients with miscellaneous tumors and 99 of 100 blood donors. One of the blood donors had low levels of Hu antibodies. The radiobinding assay detected Hu antibodies in 45 of 191 (24%) patients with SCLC, in comparison with immunofluorescence and immuno dot blot, where only 34 of 191 (18%) were detected. All patients with Hu antibodies detected by immunofluorescence and immuno dot-blot were also positive by the radioimmunobinding assay. The results demonstrate that this assay is very specific and sensitive for the detection of Hu antibodies.


Termolecular ion-molecule reactions in Titan's atmosphere. IV. A search made at up to 1 micron in pure hydrocarbons.
The results of a study of ion-molecule reactions occurring in pure methane, acetylene, ethylene, ethane, propyne, propene, propane, and diacetylene at pressures up to 40 microns of pressure are reported. A variety of experimental methods are used: The standard double resonance in an ICR, for determination of the precursor ions and the modulated double resonance ejection in an ICR, for the determination of the daughter ions. The FA-SIFT technique was used for validation and examination of termolecular reactions with rate coefficients that are less than 10(-26) cm(6) s(-1). An extensive database of reaction kinetics already exists for many of these reactions. The main point of this study was the determination of the accuracy of this database and to search for any missing reactions and reaction channels that may have been omitted from earlier investigations. A specific objective of this work was to extend the study to the highest pressures possible to find out if there were any important termolecular reaction channels occurring. A new approach was used here. In the pure hydrocarbon gases the mass spectra were followed as a function of the pressure changes of the gas. An initial guess was first made using the current literature as a source of the reaction kinetics that were expected. A model of the ion abundances was produced from the solution of the partial differential equations in terms of reaction rate coefficients and initial abundances. The experimental data was fitted to the model for all of the pressures by a least squares minimization to the reaction rate coefficients and initial abundances. The reaction rate coefficients obtained from the model were then compared to the literature values. Several new channels and reactions were discovered when the modeled fits were compared to the actual data. This is all explained in the text and the implications of these results are discussed for the Titan atmosphere.


Genome Sequence of the Probiotic Strain Lactobacillus rhamnosus (Formerly Lactobacillus casei) LOCK900.
Lactobacillus rhamnosus LOCK900 fulfills the criteria required for probiotic strains. In this study, we report a whole-genome sequence of this isolate and compare it with other L. rhamnosus complete genome sequences already published.


Launching of the yeast 20 s RNA narnavirus by expressing the genomic or antigenomic viral RNA in vivo.
20 S RNA virus is a persistent positive strand RNA virus found in Saccharomyces cerevisiae. The viral genome encodes only its RNA polymerase, p91, and resides in the cytoplasm in the form of a ribonucleoprotein complex with p91. We succeeded in generating 20 S RNA virus in vivo by expressing, from a vector, genomic strands fused at the 3'-ends to the hepatitis delta virus antigenomic ribozyme. Using this launching system, we analyzed 3'-cis-signals present in the genomic strand for replication. The viral genome has five-nucleotide inverted repeats at both termini (5'-GGGGC... GCCCC-OH). The fifth G from the 3'-end was dispensable for replication, whereas the third and fourth Cs were essential. The 3'-terminal and penultimate Cs could be eliminated or modified to other nucleotides; however, the generated viruses recovered these terminal Cs. Furthermore, extra nucleotides added at the viral 3'-end were eliminated in the launched viruses. Therefore, 20 S RNA virus has a mechanism(s) to maintain the correct size and sequence of the viral 3'-end. This may contribute to its persistent infection in yeast. We also succeeded in generating 20 S RNA virus similarly from antigenomic strands provided active p91 was supplied from a second vector in trans. Again, a cluster of four Cs at the 3'-end in the antigenomic strand was essential for replication. In this work, we also present the first conclusive evidence that 20 S and 23 S RNA viruses are independent replicons.


Neoactinobaculum massilliense gen. nov., a new genesis and Pseudopropionibacterium massiliense sp. nov., a new bacterium isolated from the human oral microbiota.
Neoactinobaculum massilliense gen. nov., strain Marseille-P6182T (= CSUR P6182) and Pseudopropionibacterium massiliense sp. nov., strain Marseille-P6184T (= CSUR P6184) are a new bacterial genus and new bacterial species belonging to the Actinobacteria phylum that have been isolated from the human oral microbiota.


Phylogeny and physiology of candidate phylum BRC1 inferred from the first complete metagenome-assembled genome obtained from deep subsurface aquifer.
Candidate bacterial phylum BRC1 has been identified in a broad range of mostly organic-rich oxic and anoxic environments through molecular analysis of microbial communities. None of the members of BRC1 have been cultivated and only a few draft genome sequences have been obtained from metagenomes or as a result of single-cell sequencing. We have reconstructed complete genome of BRC1 bacterium, BY40, from metagenome of the microbial community of a deep subsurface thermal aquifer in the Tomsk Region of the Western Siberia, Russia, and used it for metabolic reconstruction and comparison with existing genomic data. Analysis of 3.3Mb genome of BY40 bacterium revealed numerous glycoside hydrolases that could enable utilization of carbohydrates, including enzymes of chitin-degradation pathway. The bacterium lacks flagellar machinery but the twitching motility is encoded. The reconstructed central metabolism revealed pathways enabling the fermentation of organic substrates, as well as their complete oxidation through aerobic and anaerobic respiration. Phylogenetic analysis using BY40 genome supported the phylum level classification of BRC1 lineage. Based on phylogenetic and genomic analyses, the novel bacterium is proposed to be classified as Candidatus Sumerlaea chitinivorans, within a candidate phylum Sumerlaeota.


Acute effects of daily step count on postprandial metabolism and resting fat oxidation: a randomized controlled trial.
To examine the effects of daily step count on same-day fat oxidation and postprandial metabolic responses to an evening high-fat mixed meal (HFMM). Ten healthy participants (5 females, 30 ± 7 yr) completed four different daily step counts-2,000 (2 K), 5,000 (5 K), 10,000 (10 K), and 15,000 (15 K) steps-on separate days in randomized order. On experimental days, participants ate the same meals and walked all steps on an indoor track at a pace of 100 steps/min in three roughly equal bouts throughout the day. After the final walking bout, participants' resting energy expenditure (REE), respiratory exchange ratio (RER), and fat oxidation rate (FATOX) were measured. Blood samples were obtained before (BL) and 30-, 60-, 90-, 120-, and 240-min following consumption of an HFMM (960 kcal; 48% fat) to measure triglycerides (i.e., postprandial lipemia; PPL), nonesterified fatty acids (NEFAs), insulin, and glucose. Two-way ANOVAs indicated condition effects where PPL was significantly higher after 2 K versus 10 K (+23 ± 8 mg/dL, P = 0.027), and NEFAs were significantly higher after 15 K versus 2 K (+86 ± 23 µmol/L; P = 0.006). No differences were found for insulin, glucose, or REE among conditions (all P > 0.124). Similarly, RER (P = 0.054; ηp2 = 0.24) and FATOX (P = 0.071; ηp2 = 0.23) were not significantly different among conditions. In young adults, 10 K steps elicited the greatest decrease in PPL, an established cardiovascular disease risk factor. NEFA levels were highest after the 15 K condition, likely due to alterations in adipose tissue lipolysis or lipoprotein lipase activity with increased activity.NEW & NOTEWORTHY This randomized controlled trial demonstrated that walking 10,000, compared with 2,000, steps/day significantly reduced postprandial lipemia (PPL), an independent predictor of cardiovascular disease (CVD) following same-day evening meal consumption. These experimental data support walking 10,000 steps/day to lower CVD risk.


Case Study of N-i Pr versus N-Mes Substituted NHC Ligands in Nickel Chemistry: The Coordination and Cyclotrimerization of Alkynes at [Ni(NHC)2 ].
A case study on the effect of the employment of two different NHC ligands in complexes [Ni(NHC)2 ] (NHC=i Pr2 ImMe 1Me , Mes2 Im 2) and their behavior towards alkynes is reported. The reaction of a mixture of [Ni2 (i Pr2 ImMe )4 (μ-(η2 : η2 )-COD)] B/ [Ni(i Pr2 ImMe )2 (η4 -COD)] B' or [Ni(Mes2 Im)2 ] 2, respectively, with alkynes afforded complexes [Ni(NHC)2 (η2 -alkyne)] (NHC=i Pr2 ImMe : alkyne=MeC≡CMe 3, H7 C3 C≡CC3 H7 4, PhC≡CPh 5, MeOOCC≡CCOOMe 6, Me3 SiC≡CSiMe3 7, PhC≡CMe 8, HC≡CC3 H7 9, HC≡CPh 10, HC≡C(p-Tol) 11, HC≡C(4-t Bu-C6 H4 ) 12, HC≡CCOOMe 13; NHC=Mes2 Im: alkyne=MeC≡CMe 14, MeOOCC≡CCOOMe 15, PhC≡CMe 16, HC≡C(4-t Bu-C6 H4 ) 17, HC≡CCOOMe 18). Unusual rearrangement products 11 a and 12 a were identified for the complexes of the terminal alkynes HC≡C(p-Tol) and HC≡C(4-t Bu-C6 H4 ), 11 and 12, which were formed by addition of a C-H bond of one of the NHC N-i Pr methyl groups to the C≡C triple bond of the coordinated alkyne. Complex 2 catalyzes the cyclotrimerization of 2-butyne, 4-octyne, diphenylacetylene, dimethyl acetylendicarboxylate, 1-pentyne, phenylacetylene and methyl propiolate at ambient conditions, whereas 1Me is not a good catalyst. The reaction of 2 with 2-butyne was monitored in some detail, which led to a mechanistic proposal for the cyclotrimerization at [Ni(NHC)2 ]. DFT calculations reveal that the differences between 1M e and 2 for alkyne cyclotrimerization lie in the energy profile of the initiation steps, which is very shallow for 2, and each step is associated with only a moderate energy change. The higher stability of 3 compared to 14 is attributed to a better electron transfer from the NHC to the metal to the alkyne ligand for the N-alkyl substituted NHC, to enhanced Ni-alkyne backbonding due to a smaller CNHC -Ni-CNHC bite angle, and to less steric repulsion of the smaller NHC i Pr2 ImMe .


eIF2-dependent and eIF2-independent modes of initiation on the CSFV IRES: a common role of domain II.
Specific interactions of the classical swine fever virus internal ribosomal entry site (IRES) with 40S ribosomal subunits and eukaryotic translation initiation factor (eIF)3 enable 43S preinitiation complexes containing eIF3 and eIF2-GTP-Met-tRNA(iMet) to bind directly to the initiation codon, yielding 48S initiation complexes. We report that eIF5B or eIF5B/eIF3 also promote Met-tRNA(iMet) binding to IRES-40S complexes, forming 48S complexes that can assemble elongation-competent ribosomes. Although 48S complexes assembled both by eIF2/eIF3- and eIF5B/eIF3-mediated Met-tRNA(iMet) recruitment were destabilized by eIF1, dissociation of 48S complexes formed with eIF2 could be out-competed by efficient subunit joining. Deletion of IRES domain II, which is responsible for conformational changes induced in 40S subunits by IRES binding, eliminated the sensitivity of 48S complexes assembled by eIF2/eIF3- and eIF5B/eIF3-mediated mechanisms to eIF1-induced destabilization. However, 48S complexes formed by the eIF5B/eIF3-mediated mechanism on the truncated IRES could not undergo efficient subunit joining, as reported previously for analogous complexes assembled with eIF2, indicating that domain II is essential for general conformational changes in 48S complexes, irrespective of how they were assembled, that are required for eIF5-induced hydrolysis of eIF2-bound GTP and/or subunit joining.


[The accumulation and degradation dynamics of cyanophycin in cyanobacteria grown in symbiotic associations with plant tissues and cells].
Five different artificial associations of cyanobacterial cells with the cells or tissues of nightshade and rauwolfia were studied. The associations grown on nitrogen-containing media produced heterocysts. Cyanobacterial cells in the associations retained their ability to take up bound nitrogen from the medium, to store it in the form of cyanophycin granules, and to use them in the process of symbiotic growth. The synthesis and degradation of cyanophycin granules in cyanobacterial cells were more active in the associations than in monocultures. In the symbiotic associations of Chlorogloeopsis fritschii ATCC 27193 with Solanum laciniatum cells and of Nostoc muscorum CALU 304 with the Rauwolfia serpentina callus, heterocysts were produced at 3- to 30-fold higher cyanophycin contents than in cyanobacterial monocultures. In contrast, in the association of N. muscorum CALU 304 with the Solanum dulcamara callus, heterocysts were produced at lower cyanophycin contents than in the N. muscorum CALU 304 monoculture. The degradation of cyanophycin granules in N. muscorum CALU 304 cells grown in associations with plant tissues or cells was subjected to mathematical analysis. The activation of cyanophycin degradation and heterocyst production in the associations N. muscorum CALU 304-R. serpentina and C. fritschii-S. laciniatum was accompanied by an enhanced synthesis of the nitrogen-containing alkaloids in plant cells. The data obtained suggest that an integrated system of nitrogen homeostasis can be formed in symbiotic associations. Depending on the growth stage of an association, its plant member can either stimulate the accumulation of bound nitrogen in vegetative cyanobacterial cells in the form of cyanophycin granules, or activate their degradation, or initiate the formation of heterocysts independently of the cyanobacterial sensory-signalling system.


Traumatic experiences in childhood and adolescence: a meta-analysis of prospective studies assessing risk for psychosis.
Evidence of the association between traumatic experiences and psychosis are uncertain with respect to temporal order, clinical outcomes and the role of the age and genetic liability. The aim of the present meta-analysis was to explore the temporal relationship between the development of psychosis and traumatic exposure using prospective studies and to examine the role of moderation factors on overall effect sizes. Studies were identified by searching Embase-Ovid, PsycINFO (EBSCO), Pubmed, Scopus, Web of Science databases, and yielded an initial total of 9016 papers, leaving finally 23 after the screening process. Three sets of meta-analyses estimated the risk of developing psychotic experiences or full clinical psychosis by having experienced maltreatment by an adult or bullying by peers or parental death, using the random-effects model. Bullying by peers (OR = 2.28 [1.64, 4.34]), maltreatment by an adult (OR = 2.20 [1.72, 2.81]) and parental death (OR = 1.24 [1.06, 1.44]) all increased the risk of psychosis. Moderator analysis showed that negative effects of bullying were detected especially in those with genetic liability for psychosis and exposure to multiple trauma types; studies with higher prevalence of males showed a stronger risk for those exposed to parental death. No significant meta-regression was found between the risk of developing a full clinical psychosis or a psychotic experience. Lack of studies hampered the results about the age of trauma occurrence. The cumulative effect of being bullied from peers and experiencing other adversities during childhood and/or adolescence, together with genetic liability for psychosis, appears to confer the highest risk for developing psychotic symptoms later in life.


OnabotulinumtoxinA in elderly patients with chronic migraine: insights from a real-life European multicenter study.
Although migraine prevalence decreases with aging, some older patients still suffer from chronic migraine (CM). This study aimed to investigate the outcome of OnabotulinumtoxinA (OBT-A) as preventative therapy in elderly CM patients. This is a post hoc analysis of real-life prospectively collected data at 16 European headache centers on CM patients treated with OBT-A over the first three treatment cycles (i.e., Cy1-3). We defined: OLD patients aged ≥ 65 years and nonOLD those < 65-year-old. The primary endpoint was the changes in monthly headache days (MHDs) from baseline to Cy 1-3 in OLD compared with nonOLD participants. The secondary endpoints were the responder rate (RR) ≥ 50%, conversion to episodic migraine (EM) and the changes in days with acute medication use (DAMs). In a cohort of 2831 CM patients, 235 were OLD (8.3%, 73.2% females, 69.6 years SD 4.7). MHDs decreased from baseline (24.8 SD 6.2) to Cy-1 (17.5 SD 9.1, p < 0.000001), from Cy-1 to Cy-2 (14.8 SD 9.2, p < 0.0001), and from Cy-2 to Cy-3 (11.9 SD 7.9, p = 0.001). DAMs progressively reduced from baseline (19.2 SD 9.8) to Cy-1 (11.9 SD 8.8, p < 0.00001), to Cy-2 (10.9 SD 8.6, p = 0.012), to Cy-3 (9.6 SD 7.4, p = 0.049). The 50%RR increased from 30.7% (Cy-1) to 34.5% (Cy-2), to 38.7% (Cy-3). The above outcome measures did not differ in OLD compared with nonOLD patients. In a population of elderly CM patients with a long history of migraine OBT-A provided a significant benefit, over the first three treatment cycles, as good as in non-old patients.