Discovery of novel hepatocyte eQTLs in African Americans.
African Americans (AAs) are disproportionately affected by metabolic diseases and adverse drug events, with limited publicly available genomic and transcriptomic data to advance the knowledge of the molecular underpinnings or genetic associations to these diseases or drug response phenotypes. To fill this gap, we obtained 60 primary hepatocyte cultures from AA liver donors for genome-wide mapping of expression quantitative trait loci (eQTL) using LAMatrix. We identified 277 eGenes and 19,770 eQTLs, of which 67 eGenes and 7,415 eQTLs are not observed in the Genotype-Tissue Expression Project (GTEx) liver eQTL analysis. Of the eGenes found in GTEx only 25 share the same lead eQTL. These AA-specific eQTLs are less correlated to GTEx eQTLs. in effect sizes and have larger Fst values compared to eQTLs found in both cohorts (overlapping eQTLs). We assessed the overlap between GWAS variants and their tagging variants with AA hepatocyte eQTLs and demonstrated that AA hepatocyte eQTLs can decrease the number of potential causal variants at GWAS loci. Additionally, we identified 75,002 exon QTLs of which 48.8% are not eQTLs in AA hepatocytes. Our analysis provides the first comprehensive characterization of AA hepatocyte eQTLs and highlights the unique discoveries that are made possible due to the increased genetic diversity within the African ancestry genome.


Integrative assessment of biomarker responses in pale chub (Zacco platypus) exposed to copper and benzo[a]pyrene.
Multi-level biomarker responses (molecular/biochemical and histological/physiological levels) were studied to assess the sublethal toxicities of copper (Cu: 1.25, 5, and 20μg/L) and benzo[a]pyrene (BaP: 0.5, 5, and 50μg/L) induced in the freshwater pale chub Zacco platypus. Except for the kidney tissues when exposed to 20μg Cu/L, no significant differences were observed at the histological or physiological levels among the treatment groups. However, various molecular and biochemical responses were observed in Z. platypus, and these responses primarily depended on exposure time. Upon Cu exposure, both DNA single-strand breaks (COMET) and metallothionein (MT) concentration significantly increased after 4 days, whereas there were no significant changes after 14 days of exposure. Both 4 and 14 days of BaP exposure induced significant increases in COMET and 7-ethoxyresorufin-O-deethylase (EROD) activity, but there was no significant difference between them. Additionally, both Cu and BaP induced acetylcholinesterase (AChE) activity only after 14 days of exposure. The current findings demonstrate that the differences in the responses of MT and EROD are associated with each chemical's particular mode of action. Biomarker responses at the molecular and biochemical levels were quantized in terms of the integrated biomarker response (IBR) index to compare the toxicities of Cu and BaP. The IBR values were well correlated with the concentrations of Cu and BaP, and the correlations were enhanced at 4 days of exposure (r(2)=0.849 and 0.945, respectively) compared with 14 days (r(2)=0. 412 and 0.634, respectively). These results suggest that the IBR index may be a useful tool for the integrative quantification of the molecular and biochemical biomarker responses in a short-term exposure to Cu and BaP.


Higher order correlations in a levitated nanoparticle phonon laser.
We present theoretical and experimental investigations of higher order correlations of mechanical motion in the recently demonstrated optical tweezer phonon laser, consisting of a silica nanosphere trapped in vacuum by a tightly focused optical beam [R. M. Pettit et al., Nature Photonics 13, 402 (2019)]. The nanoparticle phonon number probability distribution is modeled with the master equation formalism in order to study its evolution across the lasing threshold. Up to fourth-order equal-time correlation functions are then derived from the probability distribution. Subsequently, the master equation is transformed into a nonlinear quantum Langevin equation for the trapped particle's position. This equation yields the non-equal-time correlations, also up to fourth order. Finally, we present experimental measurements of the phononic correlation functions, which are in good agreement with our theoretical predictions. We also compare the experimental data to existing analytical Ginzburg-Landau theory where we find only a partial match.


Primary pulmonary hypertension.
Primary pulmonary hypertension is a rare disease of unknown etiology, which mainly affects people in the third and fourth decades. Although the clinical cause is highly variable and spontaneous regression has been described, survival rates are usually poor, averaging only three years after diagnosis. Treatment with vasodilator agents, especially calcium channel blockers and prostaglandins, usually improves symptoms, but have not been shown to improve chances for survival. In a few selected patients who continue to deteriorate despite medical treatment, lung transplantation may be the only option for improving function and survival.


Bile Acid Diarrhea and NAFLD: Shared Pathways for Distinct Phenotypes.
Irritable bowel syndrome with diarrhea (IBS-D) and NAFLD are both common conditions that may be influenced by shared pathways of altered bile acid (BA) signaling and homeostatic regulation. Pathophysiological links between IBS-D and altered BA metabolism include altered signaling through the ileal enterokine and fibroblast growth factor 19 (FGF19) as well as increased circulating levels of 7α-hydroxy-4-cholesten-3-one, a metabolic intermediate that denotes increased hepatic BA production from cholesterol. Defective production or release of FGF19 is associated with increased BA production and BA diarrhea in some IBS-D patients. FGF19 functions as a negative regulator of hepatic cholesterol 7α-hydroxylase; therefore, reduced serum FGF19 effectively de-represses hepatic BA production in a subset of IBS-D patients, causing BA diarrhea. In addition, FGF19 modulates hepatic metabolic homeostatic response signaling by means of the fibroblast growth factor receptor 4/klotho beta receptor to activate cascades involved in hepatic lipogenesis, fatty acid oxidation, and insulin sensitivity. Emerging evidence of low circulating FGF19 levels in subsets of patients with pediatric and adult NAFLD demonstrates altered enterohepatic BA homeostasis in NAFLD. Conclusion: Here we outline how understanding of shared pathways of aberrant BA homeostatic signaling may guide targeted therapies in some patients with IBS-D and subsets of patients with NAFLD.


The influence of emotional target cues on prospective memory performance in depression.
This study investigated the impact of emotional valence on event-based prospective memory performance in depression. Thirty individuals with depression and 29 healthy adults performed a prospective memory task in which the emotional valence of the prospective targets was manipulated (positive, neutral, negative). Collapsed across all valence conditions, healthy adults outperformed individuals with depression in the prospective memory task. This effect was qualified by planned contrasts indicating that the two groups only differed when responding to positively valenced cues, reflecting a positivity effect in healthy adults. These data are in line with previous research, which shows that healthy participants better remember positively valenced cues, but are the first to show an absence of this effect in those with depression.


Dichroic rugate filters based on birefringent porous silicon.
Rugate filters made of anisotropically nanostructured birefringent silicon have been fabricated and studied by polarization-resolved transmission measurements. Electrochemical etching of a (110) oriented Si wafer results in porous silicon layers which exhibit a strong in-plane birefringence. We demonstrate that a sinusoidal refractive index variation of birefringent porous silicon combined with index-matching layers and apodization results in a dichroic rugate filter having a stop-band dependent on the polarization direction of the incident light without higher-order harmonics and sidelobes. We also demonstrate that the combination of different dichroic rugate filters allow us to realize filters with more complex properties in a single preparation step.


Crystal structure of the cytoplasmic phosphatase and tensin homolog (PTEN)-like region of Ciona intestinalis voltage-sensing phosphatase provides insight into substrate specificity and redox regulation of the phosphoinositide phosphatase activity.
Ciona intestinalis voltage-sensing phosphatase (Ci-VSP) has a transmembrane voltage sensor domain and a cytoplasmic region sharing similarity to the phosphatase and tensin homolog (PTEN). It dephosphorylates phosphatidylinositol 4,5-bisphosphate and phosphatidylinositol 3,4,5-trisphosphate upon membrane depolarization. The cytoplasmic region is composed of a phosphatase domain and a putative membrane interaction domain, C2. Here we determined the crystal structures of the Ci-VSP cytoplasmic region in three distinct constructs, wild-type (248-576), wild-type (236-576), and G365A mutant (248-576). The crystal structure of WT-236 and G365A-248 had the disulfide bond between the catalytic residue Cys-363 and the adjacent residue Cys-310. On the other hand, the disulfide bond was not present in the crystal structure of WT-248. These suggest the possibility that Ci-VSP is regulated by reactive oxygen species as found in PTEN. These structures also revealed that the conformation of the TI loop in the active site of the Ci-VSP cytoplasmic region was distinct from the corresponding region of PTEN; Ci-VSP has glutamic acid (Glu-411) in the TI loop, orienting toward the center of active site pocket. Mutation of Glu-411 led to acquirement of increased activity toward phosphatidylinositol 3,5-bisphosphate, suggesting that this site is required for determining substrate specificity. Our results provide the basic information of the enzymatic mechanism of Ci-VSP.


Neural constraints on learning.
Learning, whether motor, sensory or cognitive, requires networks of neurons to generate new activity patterns. As some behaviours are easier to learn than others, we asked if some neural activity patterns are easier to generate than others. Here we investigate whether an existing network constrains the patterns that a subset of its neurons is capable of exhibiting, and if so, what principles define this constraint. We employed a closed-loop intracortical brain-computer interface learning paradigm in which Rhesus macaques (Macaca mulatta) controlled a computer cursor by modulating neural activity patterns in the primary motor cortex. Using the brain-computer interface paradigm, we could specify and alter how neural activity mapped to cursor velocity. At the start of each session, we observed the characteristic activity patterns of the recorded neural population. The activity of a neural population can be represented in a high-dimensional space (termed the neural space), wherein each dimension corresponds to the activity of one neuron. These characteristic activity patterns comprise a low-dimensional subspace (termed the intrinsic manifold) within the neural space. The intrinsic manifold presumably reflects constraints imposed by the underlying neural circuitry. Here we show that the animals could readily learn to proficiently control the cursor using neural activity patterns that were within the intrinsic manifold. However, animals were less able to learn to proficiently control the cursor using activity patterns that were outside of the intrinsic manifold. These results suggest that the existing structure of a network can shape learning. On a timescale of hours, it seems to be difficult to learn to generate neural activity patterns that are not consistent with the existing network structure. These findings offer a network-level explanation for the observation that we are more readily able to learn new skills when they are related to the skills that we already possess.


Granule cells perform frequency-dependent pattern separation in a computational model of the dentate gyrus.
Mnemonic discrimination (MD) may be dependent on oscillatory perforant path input frequencies to the hippocampus in a "U"-shaped fashion, where some studies show that slow and fast input frequencies support MD, while other studies show that intermediate frequencies disrupt MD. We hypothesize that pattern separation (PS) underlies frequency-dependent MD performance. We aim to study, in a computational model of the hippocampal dentate gyrus (DG), the network and cellular mechanisms governing this putative "U"-shaped PS relationship. We implemented a biophysical model of the DG that produces the hypothesized "U"-shaped input frequency-PS relationship, and its associated oscillatory electrophysiological signatures. We subsequently evaluated the network's PS ability using an adapted spatiotemporal task. We undertook systematic lesion studies to identify the network-level mechanisms driving the "U"-shaped input frequency-PS relationship. A minimal circuit of a single granule cell (GC) stimulated with oscillatory inputs was also used to study potential cellular-level mechanisms. Lesioning synapses onto GCs did not impact the "U"-shaped input frequency-PS relationship. Furthermore, GC inhibition limits PS performance for fast frequency inputs, while enhancing PS for slow frequency inputs. GC interspike interval was found to be input frequency dependent in a "U"-shaped fashion, paralleling frequency-dependent PS observed at the network level. Additionally, GCs showed an attenuated firing response for fast frequency inputs. We conclude that independent of network-level inhibition, GCs may intrinsically be capable of producing a "U"-shaped input frequency-PS relationship. GCs may preferentially decorrelate slow and fast inputs via spike timing reorganization and high frequency filtering.


Orofacial indices: a study in 240 Nigerian children.
This paper seeks to establish normal orofacial indices in Nigerian children under the age of 12. The children were grouped according to age and sex. The indices studied were: mouth width/upper vermilion are with lip relaxed; coefficient of upper lip curvature; upper lip height/mouth width; height of cutaneous upper lip/upper lip height; height of upper vermilion/upper lip height; upper lip elasticity; and size of oral aperture. The indices varied with age. Except for upper lip elasticity in 10-11-year-old children, there were no significant differences in orofacial indices in males and females. The coefficient of upper lip curvature which is an index that measure the curve or protrusion of the upper lip, was maximum in two or three-year-olds in both sexes. The orofacial indices calculated from lip dimensions could provide important baseline information for surgical repair or reconstruction of the orofacial region in black children.


Strong electronic coupling dominates the absorption and fluorescence spectra of covalently bound BisBODIPYs.
The absorption spectrum of a representative BisBODIPY molecule is investigated using high-level quantum chemical methodology; the results are compared with experimental data. The S1 and S2 excited states are examined in detail to illuminate and to understand the electronic coupling between them. With the help of model systems in which the distance between the BODIPY monomers is increased or in which the dihedral angle between the subunits is changed, the electronic coupling is quantified, and its influence on energetics and oscillator strengths is highlighted. For the explanation of the experimental spectrum, orbital interaction effects are found to be important. Because of the large experimental Stokes shift of BisBODIPY, the nature of the emissive state is investigated and found to remain C2 symmetric as the ground state, and no localization of the excitation on one BODIPY subunit occurs. The excitonic coupling is in BisBODIPY still larger than the geometry relaxation energy, which explains the absence of a pseudo-Jahn-Teller effect.


A New Method for Detection of Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii Using a Novel Chromogenic Agar.
Arcobacter species are gram-negative rods that have been implicated in food- and waterborne illness. Although various cultural isolation methods have been proposed, the current procedures are unable to fully suppress the growth of background microbiota present in food samples, which inhibits Arcobacter isolation. The purpose of this study was to develop a selective enrichment broth and chromogenic plating medium to detect three Arcobacter species that have been recognized as emerging foodborne pathogens: Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii. The developed Nguyen-Restaino-Juárez (NRJ) Arcobacter detection system consists of a selective enrichment broth (NRJ-B) and a selective-differential plating medium (NRJ-M). The protocol of the detection method was determined by evaluating the growth of A. butzleri, A. cryaerophilus, and A. skirrowii under various temperatures (30, 35, and 42°C) and incubation (aerobic, microaerophilic, and anaerobic) conditions. Additionally, 47 Arcobacter strains and 39 non-Arcobacter strains were tested in inclusivity and exclusivity evaluations of NRJ-B and NRJ-M. Overall, the study determined that the optimal growth conditions of Arcobacter species using the NRJ Arcobacter detection system were aerobic incubation at 30°C. NRJ-B supported good growth of A. butzleri, A. cryaerophilus, and A. skirrowii while effectively suppressing the growth of non-Arcobacter strains after 48 h. Furthermore, NRJ-M yielded 97.8% inclusivity and 100.0% exclusivity using the tested strains and resulted in salmon-pigmented Arcobacter colonies (1.0 to 1.5 mm in diameter) after 72 h. The novel protocol is the first to develop a chromogenic plating medium for the isolation of Arcobacter species. This simple and accurate test method would greatly contribute to understanding the distribution of pathogenic Arcobacter species in food samples.


Structural elucidation of an asparagine-linked oligosaccharide from the hyperthermophilic archaeon, Archaeoglobus fulgidus.
The genome of the hyperthermophilic archaeon, Archaeoglobus fulgidus, contains three paralogous AglB genes that encode oligosaccharyltransferase (OST) proteins. The OST enzymes catalyze the transfer of an oligosaccharide chain from lipid-linked oligosaccharides (LLO) to asparagine residues in proteins. The detergent-solubilized membrane fractions prepared from cultured A. fulgidus cells contain both OST and LLO. The addition of a peptide containing the glycosylation sequon produced oligosaccharide chains attached to a structurally defined peptide. To facilitate the NMR analysis, the cells were grown in rich medium supplemented with (13)C-glucose, to label the LLOs metabolically. The MS analysis of the glycopeptide revealed that the glucose and galactose residues were nearly fully (13)C-labeled, but the mannose residues were fractionally labeled with about 20% efficiency. An immunodetection experiment revealed that the longest AglB paralog (AfAglB-L) was expressed in the membrane fractions under our cell culture conditions, while the other two shorter AglB paralogs (AfAglB-S1 and AfAglB-S2) were not. Thus, the oligosaccharide chain analyzed in this study was the product of AfAglB-L. The N-glycan consists of eight hexose residues, as follows: The α1,3-linked glucose is an optional residue branching from the distal mannose residue. The MS analysis of the minor HPLC peak of the in vitro oligosaccharyl transfer products also revealed an optional sulfate modification on the glucose residue directly linked to the Asn residue. The present data will be useful for structural and functional studies of the N-glycosylation system of A. fulgidus.


Gating of human ClC-2 chloride channels and regulation by carboxy-terminal domains.
Eukaryotic ClC channels are dimeric proteins with each subunit forming an individual protopore. Single protopores are gated by a fast gate, whereas the slow gate is assumed to control both protopores through a cooperative movement of the two carboxy-terminal domains. We here study the role of the carboxy-terminal domain in modulating fast and slow gating of human ClC-2 channels, a ubiquitously expressed ClC-type chloride channel involved in transepithelial solute transport and in neuronal chloride homeostasis. Partial truncation of the carboxy-terminus abolishes function of ClC-2 by locking the channel in a closed position. However, unlike other isoforms, its complete removal preserves function of ClC-2. ClC-2 channels without the carboxy-terminus exhibit fast and slow gates that activate and deactivate significantly faster than in WT channels. In contrast to the prevalent view, a single carboxy-terminus suffices for normal slow gating, whereas both domains regulate fast gating of individual protopores. Our findings demonstrate that the carboxy-terminus is not strictly required for slow gating and that the cooperative gating resides in other regions of the channel protein. ClC-2 is expressed in neurons and believed to open at negative potentials and increased internal chloride concentrations after intense synaptic activity. We propose that the function of the ClC-2 carboxy-terminus is to slow down the time course of channel activation in order to stabilize neuronal excitability.


Adopting the rapamycin trapping assay to track the trafficking of murine MHC class I alleles, H-2K(b).
In mammalian cells, the quality control (QC) of properly folded proteins is monitored in the early secretory pathway, particularly in the endoplasmic reticulum (ER). Several proteins, including our protein of interest, major histocompatibility complex class I (MHC class I), can bypass the first line of ER-QC and reside in post-ER compartments in an unfolded form. Such forms entail both monomeric and dimeric structures that are devoid of peptides and thus cannot fulfill the immunological function of antigen presentation at the cell surface. MHC class I structures become mature and properly folded once loaded with the appropriate peptides in the framework of the peptide loading complex (PLC). Despite the flood of information on the diverse trafficking behavior of different MHC class I alleles, there is still controversy on the actual trajectory followed by improperly folded murine MHC class I alleles, namely H-2Kb. In this study, we employ an in vitro rapamycin trapping assay, live cell imaging, and a biochemical COPII budding approach to further investigate the trafficking of H-2Kb beyond the level of the ER. We confirm the egress of H-2Kb in an unfolded form to a post-ER compartment from where they can cycle back to the ER. Deciphering the exact identity of the post-ER compartment by laser scanning microscopy did not only point to the existence of the ERGIC and cis-Golgi compartments as residency areas for unfolded proteins, but also to the involvement of an addional compartment, that lies in close proximity and possesses high resemblance to the aforementioned compartments. Interestingly, we were capable of showing using the same rapamycin trapping assay that H-2Kb can undergo a potential maturation event during their cycling; this is attained upon addition of peptides and trapping of accumulated post-ER molecules at the cell surface. Our findings deepen the understanding of H-2Kb trafficking outside the ER and pave the way to decipher the role and the trafficking of certain PLC chaperones, such as tapasin, throughout H-2K(b) post-ER QC. Finally, we demonstrate the plausible usage of the rapamycin assay to assess the trafficking of defected proteins especially in diseases and under therapeutic studies.


Prevalence and predictors of Sleep Apnea in Atrial Fibrillation patients.
Obstructive Sleep Apnea Syndrome (OSAS) is a common but often under diagnosed condition that constitutes a real public health problem. The prevalence of OSAS in atrial fibrillation (AF) patients ranges from 32 to 85% according to recent studies. To determine the prevalence and severity of OSAS in a population followed for non-valvular AF (NVAF), and to identify OSAS's predictive factors in this population. A cross-sectional study was conducted, including successive patients followed for NVAF. Patients with a known history of OSAS were not included. All patients had a clinical screening for symptoms suggestive of OSAS and underwent an ambulatory sleep study. We included 100 patients in this study. OSAS was detected in 90% of NVAF patients. The average apnea-hypopnea index (AHI) was 21.6 ± 13.6 e/h. OSAS was mild in 32%, moderate in 27% and severe in 31% of cases. Clinical characteristics were comparable between patients with and without OSAS. The multivariate analytical study concluded that independent predictive factors of OSAS in AF patients were an age greater than 61 years (p=0.029), AF duration more than 2 years (p=0.04) and snoring (p=0.01). OSAS was diagnosed in 90% of NVAF patients. Thus, a systematic screening for OSAS in AF patients may be a relevant approach given its high prevalence in this population and its therapeutic implications. Snoring, age greater than 61 years, and AF duration more than 2 years were independently associated with OSAS in our study.


Direct electron transfer catalysed by enzymes: application for biosensor development.
The ability to catalyse an electrode reaction via direct (mediatorless) electron transfer has been demonstrated for a number of redox enzymes. In the case of mediatorless electron transfer, the electron is transferred directly from the electrode to the substrate molecule via the active site of the enzyme, or vice versa. The electron itself is the second substrate for the reaction. An important point characterizing bioelectrocatalysis is the catalytic removal of the reaction over-voltage. Therefore the enzyme attached to the electrode is able to catalyse electrode reaction and forms a 'molecular transducer'. The substrate can be detected by potentiometric measurement of the removal of reaction over-voltage. The enzyme laccase is able to catalyse the reaction of oxygen electroreduction. Therefore a laccase molecular layer attached to the electrode surface forms an oxygen transducer. The formation of the layer results in a change of the electrocatalytic feature of the electrode. Laccase label coupled with either ligand or receptor allows the detection of ligand-receptor complex formation/dissociation on the electrode surface. The detection is virtually reagentless. The substrates for the reaction are molecular oxygen and the electron itself. Numerous reagentless immunosensors of different formats (competitive, displacement and sandwich) have been developed, as well as the reagentless detection system for immunofiltration/immunochromatography.


Honeybees (Apis mellifera) learn to discriminate the smell of organic compounds from their respective deuterated isotopomers.
The understanding of physiological and molecular processes underlying the sense of smell has made considerable progress during the past three decades, revealing the cascade of molecular steps that lead to the activation of olfactory receptor (OR) neurons. However, the mode of primary interaction of odorant molecules with the OR proteins within the sensory cells is still enigmatic. Two different concepts try to explain these interactions: the 'odotope hypothesis' suggests that OR proteins recognize structural aspects of the odorant molecule, whereas the 'vibration hypothesis' proposes that intra-molecular vibrations are the basis for the recognition of the odorant by the receptor protein. The vibration hypothesis predicts that OR proteins should be able to discriminate compounds containing deuterium from their common counterparts which contain hydrogen instead of deuterium. This study tests this prediction in honeybees (Apis mellifera) using the proboscis extension reflex learning in a differential conditioning paradigm. Rewarding one odour (e.g. a deuterated compound) with sucrose and not rewarding the respective analogue (e.g. hydrogen-based odorant) shows that honeybees readily learn to discriminate hydrogen-based odorants from their deuterated counterparts and supports the idea that intra-molecular vibrations may contribute to odour discrimination.


Development of skeletal systems for ICRP pediatric mesh-type reference computational phantoms.
In 2016, the International Commission on Radiological Protection (ICRP) launched Task Group 103 (TG 103) for the explicit purpose of developing a new generation of adult and pediatric reference computational phantoms, named 'mesh-type reference computational phantoms (MRCPs)', that can overcome the limitations of voxel-type reference computational phantoms (VRCPs) of ICRPPublications 110and143due to their finite voxel resolutions and the nature of voxel geometry. After completing the development of the adult MRCPs, TG 103 has started the development of pediatric MRCPs comprising 10 phantoms (male and female versions of the reference newborn, 1-year-old, 5-year-old, 10-year-old, and 15-year-old). As part of the TG 103 project, within the present study, the skeletal systems, one of the most important and complex organ systems of the body, were developed for each phantom age and sex. The developed skeletal systems, while closely preserving the original bone topology of the pediatric VRCPs, present substantial improvements in the anatomy of complex and/or small bones. In order to investigate the dosimetric impact of the developed skeletons, the average absorbed doses and the specific absorbed fractions for radiosensitive skeletal tissues (i.e. active marrow and bone endosteum) were computed for some selected external and internal exposure cases, which were then compared with those calculated with the skeletons of pediatric VRCPs. The comparison result showed that the dose values of the pediatric MRCPs were generally similar to those of the pediatric VRCPs for highly penetrating radiations (e.g. photons >200 keV); however, for weakly penetrating radiations (e.g. photons ⩽200 keV and electrons), significant differences up to a factor of 140 were observed.


Insight into the practical performance of RT-PCR testing for SARS-CoV-2 using serological data: a cohort study.
Virological detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) through RT-PCR has limitations for surveillance. Serological tests can be an important complementary approach. We aimed to assess the practical performance of RT-PCR-based surveillance protocols and determine the extent of undetected SARS-CoV-2 infection in Shenzhen, China. We did a cohort study in Shenzhen, China and attempted to recruit by telephone all RT-PCR-negative close contacts (defined as those who lived in the same residence as, or shared a meal, travelled, or socially interacted with, an index case within 2 days before symptom onset) of all RT-PCR-confirmed cases of SARS-CoV-2 detected since January, 2020, via contact tracing. We measured anti-SARS-CoV-2 antibodies in serum samples from RT-PCR-negative close contacts 2-15 weeks after initial virological testing by RT-PCR, using total antibody, IgG, and IgM ELISAs. In addition, we did a serosurvey of volunteers from neighbourhoods with no reported cases, and from neighbourhoods with reported cases. We assessed rates of infection undetected by RT-PCR, performance of RT-PCR over the course of infection, and characteristics of individuals who were seropositive on total antibody ELISA but RT-PCR negative. Between April 12 and May 4, 2020, we enrolled and collected serological samples from 2345 (53·0%) of 4422 RT-PCR-negative close contacts of cases of RT-PCR-confirmed SARS-CoV-2. 1175 (50·1%) of 2345 were close contacts of cases diagnosed in Shenzhen with contact tracing details, and of these, 880 (74·9%) had serum samples collected more than 2 weeks after exposure to an index case and were included in our analysis. 40 (4·5%) of 880 RT-PCR-negative close contacts were positive on total antibody ELISA. The seropositivity rate with total antibody ELISA among RT-PCR-negative close contacts, adjusted for assay performance, was 4·1% (95% CI 2·9-5·7), which was significantly higher than among individuals residing in neighbourhoods with no reported cases (0·0% [95% CI 0·0-1·1]). RT-PCR-positive individuals were 8·0 times (95% CI 5·3-12·7) more likely to report symptoms than those who were RT-PCR-negative but seropositive, but both groups had a similar distribution of sex, age, contact frequency, and mode of contact. RT-PCR did not detect 48 (36% [95% CI 28-44]) of 134 infected close contacts, and false-negative rates appeared to be associated with stage of infection. Even rigorous RT-PCR testing protocols might miss a substantial proportion of SARS-CoV-2 infections, perhaps in part due to difficulties in determining the timing of testing in asymptomatic individuals for optimal sensitivity. RT-PCR-based surveillance and control protocols that include rapid contact tracing, universal RT-PCR testing, and mandatory 2-week quarantine were, nevertheless, able to contain community spread in Shenzhen, China. The Bill & Melinda Gates Foundation, Special Foundation of Science and Technology Innovation Strategy of Guangdong Province, and Key Project of Shenzhen Science and Technology Innovation Commission.


[Problems of medical parasitology].
Situation on parasitic disease in Russia remains complex. Reduction of parasitology personnel had a negative impact on the quality of epidemiological control in the field of parasitic diseases and resulted in a decrease of awareness of physicians of therapeutic-prophylaxis institutions. The situation was aggravated by a lack of anti-malaria preparations and insufficient specter of anti-helminthic drugs. Uncontrolled increase of the number of domestic and stray dogs in cities was the reason for increase of morbidity by helminthoses and zoonoses--toxocarosis and dirofilariasis. Emergence in the south of Krasnodar Region of effective carriers Aedes aegypti, Aedes albopictus became a serious threat to biological safety of the country. These mosquitos are effective carriers of causative agents of mosquito viral fevers: yellow, Dengue, Chikungunya et al.


Synthesis and characterization of novel organo-hydrogel based agar, glycerol and peppermint oil as a natural drug carrier/release material.
The very recent Covid-19 pandemic has made the need to understand biocompatible polymers as support material in drug delivery systems and controlled release clearer, especially for organo-hydrogels. This study aims to synthesize various new polymeric materials called gels, hydrogels, and organo-hydrogels according to the monomer used and to investigate their use as drug release systems. The agar-glycerol (AG) pair was used to synthesize the polymers, N, N, methylene bisacrylamide (MBA, m) and glutaraldehyde (GA, g) were used as cross-linkers and peppermint oil (PmO) was included to obtain the organo-hydrogels. Therefore, one AG gel and two p (AG-m) and p (GA-g) hydrogels were synthesized within the scope of the study. Six different organo-hydrogels based on p(AG-m-PmO) or p (AG-g-PmO) were also synthesized by varying the amount of peppermint oil. Paracetamol and carboplatin were selected as the sample drugs. Synthesized gels, hydrogels and organo-hydrogels were characterized by FTIR and SEM analysis. Additionally, swelling behaviors of the synthesized gels were investigated in different media (ID water, tap water, ethanol, acetone, ethanol/ID water (1:1), acetone/ID water (1:1) and gasoline) and at different pHs. Moreover, it was determined that organo-hydrogels were blood compatible and had antioxidant properties based on hemolysis, blood clotting and antioxidant analysis. Therefore, the release of paracetamol (a known antipyretic-painkiller, recommended and used in the treatment of Covid-19) and carboplatin (widely used in cancer treatment) were studied. Evidently, as the amount of PMO oil increases, the -OH groups in organo-hydrogels will increase and the chemical and physical bonding rates will increase; therefore it was observed that increasing peppermint oil in the organo-hydrogels structure to 0.3 mL stimulated the release of the drugs. For instance, maximum paracetamol release amount from p(AG-g-PmO) and p(AG-m-PmO) organo-hydrogels was calculated to be 72.3% at pH 7.4 and 69.8% at pH 2.0, respectively. The maximum carboplatin release amount from p(AG-g-PmO) and p(AG-m-PmO) organo-hydrogels was calculated to be 99.7% at pH 7.4 and 100% at pH 7.4, respectively. It was concluded that the synthesized organo-hydrogels might easily be used as drug carrier and controlled drug release materials.


Identification of a 150-kilodalton polypeptide that copurifies with yeast TFIIIC and binds specifically to tRNA genes.
The transcription in vitro of eucaryotic tRNA genes by RNA polymerase III requires two transcription factors, designated TFIIIB and TFIIIC. One of the critical functions of TFIIIC in the transcription of tRNA genes is that it interacts directly and specifically with the two internal promoter elements of these genes. We have partially purified Saccharomyces cerevisiae TFIIIC by chromatography on Bio-Rex 70, DEAE-cellulose, and phosphocellulose resins. A 150-kilodalton (kDa) DNA-binding polypeptide copurified with TFIIIC activity. This 150-kDa protein coeluted with the DNA-binding activity of TFIIIC after rechromatography of TFIIIC on phosphocellulose and its elution with a linear salt gradient. The stable and high-affinity interaction of this protein with tRNA genes was demonstrated by the maintenance of a protein-DNA complex under conditions of high ionic strength. Finally, we showed by two criteria that the interaction of this protein with tRNA genes was specific. First, the protein-DNA complex was competed with only by DNA-containing tRNA genes; second, the protein preferentially bound to DNA fragments containing a tRNA gene. These results strongly suggest that the DNA-binding domain of the yeast TFIIIC is contained within this 150-kDa polypeptide.


Metabolite content profiling of bottlenose dolphin exhaled breath.
Changing ocean health and the potential impact on marine mammal health are gaining global attention. Direct health assessments of wild marine mammals, however, is inherently difficult. Breath analysis metabolomics is a very attractive assessment tool due to its noninvasive nature, but it is analytically challenging. It has never been attempted in cetaceans for comprehensive metabolite profiling. We have developed a method to reproducibly sample breath from small cetaceans, specifically Atlantic bottlenose dolphins (Tursiops truncatus). We describe the analysis workflow to profile exhaled breath metabolites and provide here a first library of volatile and nonvolatile compounds in cetacean exhaled breath. The described analytical methodology enabled us to document baseline compounds in exhaled breath of healthy animals and to study changes in metabolic content of dolphin breath with regard to a variety of factors. The method of breath analysis may provide a very valuable tool in future wildlife conservation efforts as well as deepen our understanding of marine mammals biology and physiology.


Construction of a hepatic stellate cells subtracted cDNA library of differentially expressed genes in normal mice and mice with Schistosomiasis japonica.
To construct a hepatic stellate cells (HSCs) subtracted cDNA library to find differentially expressed genes in normal mice and mice infected with Schistosoma japonicum (S. japonicum). Suppression subtractive hybridization (SSH) was used. The cDNA fragments of normal mouse were compared to those of schistosoma-infected mice to find differentially expressed genes. Then differentially expressed cDNA fragments were directly inserted into T/A cloning vector to set up the subtractive library. Amplification of the library was carried out with transformation of DH5alpha. The amplified library contained more than 400 positive bacterial clone, which were then hybridized with forward and backward subtracted probes for differential screening. One hundred positive bacterial clones were randomly selected for sequencing and BLAST analysis. Finally, virtual Northern Blot confirmed such differential expression. The subtracted cDNA library of differentially expressed genes of HSCs was constructed successfully, the library is efficient and lays foundation for screening and cloning new and specific genes of schistosomiasis.


Wave2Vec: Vectorizing Electroencephalography Bio-Signal for Prediction of Brain Disease.
Interest in research involving health-medical information analysis based on artificial intelligence, especially for deep learning techniques, has recently been increasing. Most of the research in this field has been focused on searching for new knowledge for predicting and diagnosing disease by revealing the relation between disease and various information features of data. These features are extracted by analyzing various clinical pathology data, such as EHR (electronic health records), and academic literature using the techniques of data analysis, natural language processing, etc. However, still needed are more research and interest in applying the latest advanced artificial intelligence-based data analysis technique to bio-signal data, which are continuous physiological records, such as EEG (electroencephalography) and ECG (electrocardiogram). Unlike the other types of data, applying deep learning to bio-signal data, which is in the form of time series of real numbers, has many issues that need to be resolved in preprocessing, learning, and analysis. Such issues include leaving feature selection, learning parts that are black boxes, difficulties in recognizing and identifying effective features, high computational complexities, etc. In this paper, to solve these issues, we provide an encoding-based Wave2vec time series classifier model, which combines signal-processing and deep learning-based natural language processing techniques. To demonstrate its advantages, we provide the results of three experiments conducted with EEG data of the University of California Irvine, which are a real-world benchmark bio-signal dataset. After converting the bio-signals (in the form of waves), which are a real number time series, into a sequence of symbols or a sequence of wavelet patterns that are converted into symbols, through encoding, the proposed model vectorizes the symbols by learning the sequence using deep learning-based natural language processing. The models of each class can be constructed through learning from the vectorized wavelet patterns and training data. The implemented models can be used for prediction and diagnosis of diseases by classifying the new data. The proposed method enhanced data readability and intuition of feature selection and learning processes by converting the time series of real number data into sequences of symbols. In addition, it facilitates intuitive and easy recognition, and identification of influential patterns. Furthermore, real-time large-capacity data analysis is facilitated, which is essential in the development of real-time analysis diagnosis systems, by drastically reducing the complexity of calculation without deterioration of analysis performance by data simplification through the encoding process.


Characterization of nutty flavor in cheddar cheese.
The objectives of this study were to determine the volatile components responsible for the sensory perception of nutty flavor in Cheddar cheese. Cheddar cheeses with and without nutty flavors were selected by descriptive sensory analysis. Volatile aroma components from Cheddar cheeses with and without nutty flavors were isolated and characterized using solvent extraction with high vacuum distillation, dynamic headspace analysis, gas chromatography-olfactometry, and gas chromatography-mass spectrometry. More than 50 aroma-active compounds were detected in Cheddar cheeses. Consistent differences were observed between nutty and not nutty Cheddar cheeses. Strecker aldehydes were detected in higher amounts in Cheddar cheeses with nutty flavors compared with Cheddar cheeses without nutty flavors. Strecker aldehydes, dimethyl sulfide, and propionic acid were evaluated in young and aged Cheddar cheese models for nutty flavor by descriptive sensory analysis. Dimethyl sulfide and propionic acid did not contribute to nutty flavor in Cheddar cheese. The addition of Strecker aldehydes to young (<4 mo old) Cheddar cheese models resulted in nutty/malty flavor perceived by sensory analysis. When Strecker aldehydes were incorporated into aged (>9 mo old) Cheddar cheese models, nutty flavor perception increased. Strecker aldehydes contribute to nutty flavor in aged Cheddar cheese.


A tale of two mechanisms: a meta-analytic approach toward understanding the autonomic basis of cardiovascular reactivity to acute psychological stress.
A series of meta-analyses was undertaken to determine the contributions of sympathetic and parasympathetic activation to cardiovascular stress reactivity. A literature search yielded 186 studies of sufficient quality that measured indices of sympathetic (n = 113) and/or parasympathetic activity (n = 73). A range of psychological stressors perturbed blood pressure and heart rate. There were comparable aggregate effects for sympathetic activation, as indexed by increased plasma epinephrine and norepinephrine, and shortened pre-ejection period and parasympathetic deactivation, as indexed by heart rate variability measures. Effect sizes varied with stress task, sex, and age. In contrast to alpha-adrenergic blockade, beta-blockade attenuated cardiovascular reactivity. Cardiovascular reactivity to acute psychological stress would appear to reflect both beta-adrenergic activation and vagal withdrawal to a largely equal extent.


Glucose increases synaptic transmission from vagal afferent central nerve terminals via modulation of 5-HT3 receptors.
Acute hyperglycemia has profound effects on vagally mediated gastrointestinal functions. We have reported recently that the release of glutamate from the central terminals of vagal afferent neurons is correlated directly with the extracellular glucose concentration. The present study was designed to test the hypothesis that 5-HT(3) receptors present on vagal afferent nerve terminals are involved in this glucose-dependent modulation of glutamatergic synaptic transmission. Whole-cell patch-clamp recordings were made from neurons of the nucleus tractus solitarius (NTS) in thin rat brainstem slices. Spontaneous and evoked glutamate release was decreased in a concentration-dependent manner by the 5-HT(3) receptor selective antagonist, ondansetron. Alterations in the extracellular glucose concentration induced parallel shifts in the ondansetron-mediated inhibition of glutamate release. The changes in excitatory synaptic transmission induced by extracellular glucose concentration were mimicked by the serotonin uptake inhibitor, fenfluramine. These data suggest that glucose alters excitatory synaptic transmission within the rat brainstem via actions on tonically active 5-HT(3) receptors, and the number of 5-HT(3) receptors on vagal afferent nerve terminals is positively correlated with the extracellular glucose concentration. These data indicate that the 5-HT(3) receptors present on synaptic connections between vagal afferent nerve terminals and NTS neurons are a strong candidate for consideration as one of the sites where glucose acts to modulate vagovagal reflexes.


Quantile-based permutation thresholds for quantitative trait loci hotspots.
Quantitative trait loci (QTL) hotspots (genomic locations affecting many traits) are a common feature in genetical genomics studies and are biologically interesting since they may harbor critical regulators. Therefore, statistical procedures to assess the significance of hotspots are of key importance. One approach, randomly allocating observed QTL across the genomic locations separately by trait, implicitly assumes all traits are uncorrelated. Recently, an empirical test for QTL hotspots was proposed on the basis of the number of traits that exceed a predetermined LOD value, such as the standard permutation LOD threshold. The permutation null distribution of the maximum number of traits across all genomic locations preserves the correlation structure among the phenotypes, avoiding the detection of spurious hotspots due to nongenetic correlation induced by uncontrolled environmental factors and unmeasured variables. However, by considering only the number of traits above a threshold, without accounting for the magnitude of the LOD scores, relevant information is lost. In particular, biologically interesting hotspots composed of a moderate to small number of traits with strong LOD scores may be neglected as nonsignificant. In this article we propose a quantile-based permutation approach that simultaneously accounts for the number and the LOD scores of traits within the hotspots. By considering a sliding scale of mapping thresholds, our method can assess the statistical significance of both small and large hotspots. Although the proposed approach can be applied to any type of heritable high-volume "omic" data set, we restrict our attention to expression (e)QTL analysis. We assess and compare the performances of these three methods in simulations and we illustrate how our approach can effectively assess the significance of moderate and small hotspots with strong LOD scores in a yeast expression data set.


Mutations in the flhD gene of Escherichia coli K-12 do not cause the reported effect on cell division.
The hetero-oligomeric FlhD/FlhC complex is a global regulator of transcription in Escherichia coli. FlhD alone, independent of FlhC, has also been reported to control when E. coli cells stop dividing and enter the stationary phase. This work is frequently cited as evidence that FlhD regulates cell division; however, our data indicate that this is not the case. The results presented here show that the previously observed phenotype is not due to the flhD locus, but is instead due to differences in the thyA alleles present in the flhD(+) and flhD(-) strains used in the original studies. We find that when the strains being compared have the same thyA allele (wild type or mutant), flhD mutations have no effect on growth.


Compressive Strength of New Glass Ionomer Cement Technology based Restorative Materials after Thermocycling and Cyclic Loading.
The objective of the study was to compare compressive strengths of two glass ionomer-based materials, with and without a light-cured, nano-filled coating, after cyclic loading and thermocycling. To determine compressive strength of new restorative materials over a longer period of time, materials were analysed under simulated conditions where cyclic loading replicated masticatory loading and thermocycling simulated thermal oscillations in the oral cavity. Four groups of samples (n=7)-(1) Equia Fil (GC, Tokyo, Japan) uncoated; (2) Equia Fil coated with Equia Coat (GC, Tokyo, Japan); (3) Equia Forte Fil (GC, Tokyo, Japan) uncoated; and (4) Equia Forte Fil coated with Equia Forte coat (GC, Tokyo, Japan)-were subjected to cyclic loading (240,000 cycles) using a chewing simulator (MOD, Esetron Smart Robotechnologies, Ankara, Turkey). Compressive strength measurements were performed according to ISO 9917-1:2007, using the universal mechanical testing machine (Instron, Lloyd, UK). Scanning electron microscope (SEM) analysis was performed after thermocycling. There were no statistically significant differences between Equia Fil and Equia Forte Fil irrespective of the coating (p<0.05), but a trend of increasing compressive strength in the coated samples was observed. Coating increases the compressive strength of Equia Fil and Equia Forte Fil, but not significantly.


Association of hemoglobin concentration and mortality in critically ill patients with severe traumatic brain injury.
The critical care management of traumatic brain injury focuses on preventing secondary ischemic injury. Cerebral oxygen delivery is dependent upon the cerebral perfusion pressure and the oxygen content of blood, which is principally determined by hemoglobin. Despite its importance to the cerebral oxygen delivery, the precise hemoglobin concentration to provide adequate oxygen delivery to injured neuronal tissue in TBI patients is controversial with limited evidence to provide transfusion thresholds. We conducted a retrospective cohort study of severe TBI patients, investigating the association between mean 7-day hemoglobin concentration and hospital mortality. Demographic, physiologic, intensive care interventions, clinical outcomes and daily hemoglobin concentrations were recorded for all patients. Patients were all cared for at a tertiary, level 1 trauma center in a mixed medical and surgical intensive unit. Patients were divided into quartiles based on their mean 7-day hemoglobin concentration: < 90 g/L, 90 - 99 g/L, 100 - 109 g/L and > 110 g/L. Multivariable log-binomial regression was used to model the association between mean daily hemoglobin concentration and hospital mortality. Two hundred seventy-three patients with traumatic brain injury were identified and 169 were included in the analysis based on inclusion/exclusion criteria. Of these, 77% of the patients were male, with a mean age of 38 (SD 17) years and a median best GCS of 6 (IQR 5 - 7). One hundred fifteen patients (68%) received a red blood cell (RBC) transfusion. In RBCs administered in the ICU, the median pre-transfusion hemoglobin was 79 g/L (IQR 73 - 85). Thirty-seven patients (22%) died in hospital. Multivariable analysis revealed that mean 7-day hemoglobin concentration < 90 g/L was independently associated with an increased risk of hospital mortality (RR 3.1, 95% CI 1.5 - 6.3, p = 0.03). Other variables associated with increased mortality on multivariable regression were insertion of external ventricular drain, age and decreased GCS. Red blood cell transfusion was not associated with mortality following multivariable adjustment. A mean 7-day hemoglobin concentration of < 90g/L is associated with increased hospital mortality in patients with severe traumatic brain injury.


Modulation of calcium signals by intracellular pH in isolated rat pancreatic acinar cells.
1. We have investigated the interactions between intracellular pH (pH1) and the intracellular free calcium concentration ([Ca2+]i) in isolated rat pancreatic acinar cells. The fluorescent dyes fura-2 and BCECF were used to measure [Ca2+]i and pHi, respectively. 2. Sodium acetate and ammonium chloride (NH4Cl) were used to acidify and alkalinize pHi, respectively. Cytosolic acidification had no effect on [Ca2+]i in resting pancreatic acinar cells, whereas cytosolic alkalinization released Ca2+ from intracellular stores. 3. Cytosolic acidification using either acetate or a CO2-HCO3(-)-buffered medium enhanced Ca2+ signals evoked by acetylcholine (ACh) and cholecystokinin (CCK). In contrast, both NH4Cl and trimethylamine (TMA) inhibited Ca2+ signals during stimulation with either ACh or CCK. This inhibitory effect was also observed in the absence of extracellular Ca2+, and was therefore not due to changes in Ca2+ entry. 4. Calcium oscillations evoked by physiological concentrations of CCK were enhanced by cytosolic acidification and inhibited by cytosolic alkalinization. 5. In order to determine the effects of pHi upon Ca2+ handling by intracellular Ca2+ stores, intraorganellar [Ca2+] was monitored using the low affinity Ca2+ indicator mag-fura-2 in permeabilized cells. Addition of NH4Cl, which is expected to alkalinize intraorganellar pH, did not alter intraorganellar [Ca2+] in permeabilized cells, suggesting that changing intraorganellar pH does not release Ca2+ from intracellular stores. Addition of NH4Cl or acetate also did not affect the rate of Ca2+ release induced by inositol 1,4,5-trisphosphate (InsP3). 6. Modification of extraorganellar ('cytosolic') pH did not affect the rate of ATP-dependent Ca2+ uptake into stores, but did modify the rate of Ca2+ release evoked by submaximal concentrations of InsP3. The rate of Ca2+ release was increased at more alkaline extraorganellar pHs. These results would suggest that manipulation of intraorganellar pH does not affect Ca2+ handling by the intracellular stores. In contrast, extraorganellar ('cytosolic') pH does affect InsP3-induced Ca2+ release from the stores. 7. In conclusion, changes in intracellular pH in pancreatic acinar cells can profoundly alter cytosolic [Ca2+]. This may shed light on earlier observations whereby cell-permeant weak acids and bases can modulate fluid secretion in epithelia.


Reform incentives to create a demand for health system reengineering.
America needs a far more efficient health care financing and delivery system than the one we have. Our present system is a serious threat to public finances and is pricing itself out of reach. At the root of the problem are incentives and organization. The present fragmented fee-for-service small practice model is filled with cost-increasing incentives. There are some relatively efficient organized delivery systems, mostly based on large multi-specialty group practices. Unfortunately, most consumers are not offered the opportunity to save money and get better care by choosing such a system. This situation presents great opportunities for improvement in performance by re-engineering the system. However, for this to happen, incentives must be fundamentally changed so that everyone is cost conscious and care is organized in accountable care systems seeking improvement.


Thrombin Generation in Patients with Atrial Fibrillation Undergoing Percutaneous Coronary Intervention.
The optimal antithrombotic treatment for patients with atrial fibrillation (AF) that undergo percutaneous coronary intervention (PCI) is controversial. Dual therapy (clopidogrel and a direct oral anticoagulant [DOAC]) is safer than triple therapy (warfarin, aspirin, and clopidogrel), while efficacy is unclear. We aimed to evaluate thrombin generation (TG) under dual and triple therapy. A noninterventional prospective trial in patients with AF undergoing PCI. Patients received 4 weeks of triple therapy with aspirin, clopidogrel, and a DOAC followed by aspirin withdrawal. TG was measured in platelet-rich plasma (PRP) and platelet-poor plasma (PPP) at 3 five to 21 points, day 1 after PCI (TIME 0), 4 weeks after PCI (TIME 1), and 2 weeks after aspirin withdrawal (TIME 2). Twenty-three patients (18 men, median age 78 years, 83% with acute coronary syndrome) were included. Endogenous thrombin potential (ETP) in PPP was high at TIME 0 compared with TIME 1 (ETP 3,178 ± 248 nM vs. 2,378 ± 222 nM, p = 0.005). These results remained consistent when measured in PRP. No significant difference in ETP was found before (TIME 1) and after aspirin withdrawal (TIME 2) although few patients had high ETP levels after stopping aspirin. TG potential is high immediately after PCI and decreases 4 weeks after PCI in patients receiving triple therapy. TG remains constant after aspirin withdrawal in most patients, suggesting that after 1 month the antithrombotic effect of dual therapy may be similar to triple therapy.


Familial Associations of Colon and Rectal Cancers With Other Cancers.
Many studies have indicated that colon and rectal cancers differ in etiology and histology. The aim of this study was to investigate whether the associations of colon and rectal cancers with any other (discordant) cancer were site specific. A novel approach was implemented in which cancer risks were analyzed in families with increasing numbers of family members diagnosed with defined cancers. The novel assumption was that, for a true familial association, the risk should increase by the number of affected family members. In separate analyses, familial risks were calculated after the exclusion of putative families with hereditary nonpolyposis colorectal cancer. The study was conducted using the Swedish Family-Cancer Database. The outcome measure was relative risk. Relative risks of colorectal cancer and colon cancer were higher when family members were diagnosed with colon cancer than when family members were diagnosed with rectal cancer (incidence rate ratio for colorectal: 1.82 (95% CI, 1.74-1.90) vs 1.61 (95% CI, 1.51-1.71); incidence rate ratio for colon: 1.92 (95% CI, 1.83-2.02) vs 1.56 (95% CI, 1.45-1.69)). Relative risks for 10 discordant cancers were increased in colon or rectal cancer families, whereas none of the relative risks differed significantly between colon and rectal cancers. After deleting hereditary nonpolyposis colorectal cancer families, the relative risks of endometrial and ovarian cancers were no longer significant. Genetic data are unavailable in the database. Our results suggested that familial risks for colon cancer were higher than risks for rectal cancer in families of patients with colorectal cancer and colon cancer. The relationships of lung cancer and nervous system cancer with colorectal cancer were site specific. The associations of colon and rectal cancers with lung cancer, myeloma, and cancer of unknown primary appeared not to point out known syndromes and may suggest involvement of a novel predisposition. See Video Abstract at http://links.lww.com/DCR/A791.


Truncation of the COOH-terminal region of the paramyxovirus SV5 fusion protein leads to hemifusion but not complete fusion.
The role of the simian virus 5 (SV5) fusion (F) protein 20 residue COOH-terminal region, thought to represent the cytoplasmic tail, in fusion activity was examined by constructing a series of COOH-terminal truncation mutants. When the altered F proteins were expressed in eukaryotic cells, by using the vaccinia virus-T7 transient expression system, all the F proteins exhibited similar intracellular transport properties and all were expressed abundantly on the cell surface. Quantitative and qualitative cell fusion assays indicated that all of the F protein COOH-terminal truncation mutants mediated lipid mixing with similar kinetics and efficiency as that of wild-type F protein. However, the cytoplasmic content mixing activity decreased in parallel with the extent of the deletion in the F protein COOH-terminal truncation mutants. These data indicate that it is possible to separate the presumptive early step in the fusion reaction, hemifusion, and the final stage of fusion, content mixing, and that the presence of the F protein COOH-terminal region is important for the final steps of fusion.


Device therapy in heart failure patients with chronic kidney disease.
Heart failure (HF) and chronic kidney disease (CKD) both carry significant risk for sudden cardiac death, hospitalization, and mortality; when combined, however, they markedly increase the risk of morbidity and mortality. Device therapies such as implantable cardioverter-defibrillators (ICDs) and cardiac resynchronization therapy (CRT) are treatments proven to have significant benefit on clinical outcomes in select patients with HF. However, the majority of studies supporting the use of these devices have limited data on patients with CKD or end-stage renal disease. In this review, we discuss the intersection of HF and CKD as it relates to progressive HF and the risk of sudden death. Although these disorders are common and have a poor prognosis, the evidence available for guiding treatment decisions for the use of ICD and CRT devices in these patients is lacking. Given this lack of clear evidence, pragmatic clinical trials and comparative effectiveness studies are needed to help identify the appropriate use of ICD and CRT devices in this high-risk population of patients with HF and CKD.


Reducing amyloid-related Alzheimer's disease pathogenesis by a small molecule targeting filamin A.
PTI-125 is a novel compound demonstrating a promising new approach to treating Alzheimer's disease (AD), characterized by neurodegeneration and amyloid plaque and neurofibrillary pathologies. We show that the toxic signaling of amyloid-β(42) (Aβ(42)) by the α7-nicotinic acetylcholine receptor (α7nAChR), which results in tau phosphorylation and formation of neurofibrillary tangles, requires the recruitment of the scaffolding protein filamin A (FLNA). By binding FLNA with high affinity, PTI-125 prevents Aβ(42)'s toxic cascade, decreasing phospho-tau and Aβ aggregates and reducing the dysfunction of α7nAChRs, NMDARs, and insulin receptors. PTI-125 prevents Aβ(42) signaling by drastically reducing its affinity for α7nAChRs and can even dissociate existing Aβ(42)-α7nAChR complexes. Additionally, PTI-125 prevents Aβ-induced inflammatory cytokine release by blocking FLNA recruitment to toll-like receptor 4, illustrating an anti-inflammatory effect. PTI-125's broad spectrum of beneficial effects is demonstrated here in an intracerebroventricular Aβ(42) infusion mouse model of AD and in human postmortem AD brain tissue.


[The iatrogenic small bowel perforation as a complication of laparoscopic cholecystectomy].
Having done 1 125 laparoscopic cholecystectomies we want to report an exceptional case of perioperative bowel lesion. The symptoms began with abdominal pain on the third postoperative day, followed by an evacuation of intestinal contents out of the Robinson-Drainage. The laparotomy showed a 4 mm lesion of the terminal ileum, which could be excised and closed. Any other complications were not recognized. The reason that caused the perforation could not be clarified. Possible mechanisms will be discussed.


A Pedigree Report of a Rare Case of Weill-Marchesani Syndrome with New Compound Heterozygous LTBP2 Mutations.
Weill-Marchesani syndrome (WMS) is an autosomal inherited connective tissue disease. Clinical manifestations include microspherophakia (MSP), high myopia, ectopia lentis, open-angle glaucoma, short stature, short fingers, joint stiffness, and (occasionally) cardiovascular defects. At present, a total of four pathogenic gene loci related to WMS have been found: ADAMTS10, ADAMTS17, FBN1, and LTBP2. The patient was a five-year-old girl whose eyesight had become progressively worse for three years before her parents brought her to the hospital. Computer optometry showed high myopia in both eyes, while a slit lamp examination found that the anterior chamber of both eyes was shallow, and the lens was in a state of dislocation (ectopia lentis). An IOLMaster examination revealed that the lens was spherical (MSP), and the lens thickness (LT) was 5.36 mm. Corneal topography showed that the angle kappa was 0.18 mm in the right eye (OD) and 0.30 mm in the left eye (OS). An intraocular pressure (IOP) (OD: 26.5 mmHg, OS: 30.6 mmHg) examination showed that the fundus cup to disc ratio was normal, but secondary glaucoma caused by lens dislocation could be considered. The IOP was maintained within a normal range using antihypertensive drugs. The patient's younger sister also had a dislocation of MSP. Gene detection showed a heterozygous mutation in the LTBP2 gene [c.3672delC:p.Thr1225fs and c.3542delT:p.Met1181fs], and a diagnosis of WMS-like syndrome was confirmed. WMS syndrome is rare, and the mutation of the LTBP2 gene has not been previously recorded in the GnomAD (Genome Aggregation Database) of East Asia. This case report provides some reference for studying the mechanism of WMS and WMS-like syndrome caused by an LTBP2 gene mutation.


Proteomic analysis of liver mitochondria of db/db mice treated with grape seed procyanidin B2.
Hepatic damage has been recognized as one of the major complications in diabetes mellitus. Our previous studies have verified that grape seed procyanidin B2 (GSPB2) played a protective effect on hepatic damage of diabetes. We used isobaric tag for relative and absolute quantitation proteomics here to identify the alterant mitochondrial protein profile in diabetic liver and to seek the protective targets of GSPB2. Proteomics found that 171 proteins were upregulated or downregulated in the liver mitochondria of diabetic group compared to the control group. Of these proteins, 61 were normalized after GSPB2 treatment. These back-regulated proteins are involved in the process of fatty acid oxidation, tricarboxylic acid cycle, oxidative phosphorylation, oxidative stress, and apoptosis. Some differentially expressed proteins were confirmed by western blotting. Our study might help to better understand the mechanism of mitochondrial dysfunction in diabetic liver damage, and provide novel targets for estimating the protective effects of GSPB2. PRACTICAL APPLICATIONS: Grape seed procyanidin B2 (GSPB2), a polyphenolic component found in red wine and grapes, has beneficial effects such as antioxidative stress, antiapoptosis, and cardiovascular protection. We used proteomics here to identify the differentially expressed mitochondrial proteins in diabetic liver after GSPB2 treatment and to seek the protective targets of GSPB2. We found that the differentially expressed proteins were involved in carbon metabolism, oxidative phosphorylation, fatty acid metabolism, citrate cycle, oxidative stress, and apoptosis. These proteins may play a key role in diabetic hepatic damage as functional proteins. Targeting these proteins including apply of GSPB2 could potentially lead to an effective treatment in the diabetic hepatic disease.


Nature of phi chi 174 linear DNA from a DNA ligase-defective host.
Linear DNAs have been prepared from phiX phage and from phiX RF II (double-stranded circular form of phiX DNA, formed during infection and nicked in one or both strands) molecules derived from infection at the restrictive temperature of Escherichia coli ts7, a host mutant with a temperature-sensitive DNA ligase activity. The linear DNA from these phages can be circularized by annealing with fragments of phiX RF DNA produced by the Haemophilus influenzae restriction nuclease. The circularization experiment indicated that the site of breakage of the linear phage DNAs is not unique nor confined to a particular region of the genome. These linear DNAs were less than 0.1% as infective as circular phage DNA. The linear, positive strand of late RF II DNA, however, is uniquely nicked in the region of the phiX genome corresponding to cistron A. Although a low level of infectivity is associated with the linear DNA derived from late RF II, this infectivity appears to be a result of the association of linear positive and linear negative strands during the infectivity assay.


Targeted expression of c-Src in epidermal basal cells leads to enhanced skin tumor promotion, malignant progression, and metastasis.
In this study, we generated transgenic mice that overexpressed either a constitutively active human c-src mutant (src(530)) or a wild-type human c-src (src(wt)) in epidermal basal cells driven by human keratin 14 (HK14) or bovine keratin 5 (BK5) promoters, respectively. HK14.src(530) transgenic mice developed severe epidermal hyperplasia and hyperkeratosis, and did not survive beyond 3 weeks of age. Four transgenic founders were obtained after injection of a BK5.src(wt) construct with variable phenotypes, and three lines (lines A-C) were established. BK5.src(wt) founder D exhibited a severe skin phenotype similar to HK14.src(530) transgenic mice and died 5 days after birth. Line C transgenic mice also exhibited significant epidermal hyperplasia and hyperkeratosis, and developed spontaneous squamous cell carcinomas (SCCs) of the skin beginning at approximately 3 months of age (70% incidence at 1 year). Mice from lines A and B did not show a marked phenotype; however, elevated human src(wt) protein in the epidermis of line B mice was clearly evident. Additional analyses of line B transgenic mice showed an enhanced responsiveness to 12-O-tetradecanoylphorbol-13-acetate-induced epidermal hyperplasia and cell proliferation. Analysis of the susceptibility of line B mice to two-stage skin carcinogenesis revealed that papillomas and SCCs arose earlier and in greater numbers compared with nontransgenic littermates. In addition, malignant conversion occurred more rapidly, and the SCCs that developed in line B transgenic mice had a greater propensity to metastasize to peripheral lymph nodes and other organs. These observations support the hypothesis that c-src plays an important role in skin tumor promotion. In addition, the data show that elevated c-src activity enhances malignant progression and metastasis in this model system.


Group B rotaviruses similar to strain CAL-1, have been circulating in Western India since 1993.
Generally, group A rotaviruses are the most common cause of paediatric diarrhoea. However, group B rotavirus, adult diarrhoea rotavirus (ADRV), was found to be involved in epidemics of severe gastroenteritis in several areas of China during 1982-1983 and had resulted in more than one million cases among adults as well as older children. Human group B rotavirus has been rarely reported outside China, but has been detected first from five adults with diarrhoea in Kolkata, India during 1997-1998 (strain CAL-1). During epidemiological studies at the National Institute of Virology (NIV) on hospitalized diarrhoea patients at Pune, India, faecal specimens from patients of >5 years age, which were negative for group A rotavirus by ELISA were tested by polyacrylamide gel electrophoresis (PAGE). We detected rotavirus RNA migration patterns similar to that of group B rotavirus in three faecal specimens from adults, two from the specimens collected in 1993 and one in 1998 from sporadic diarrhoea cases. RT-PCR was carried out using primers derived from gene 8 which codes for the NS2 protein, followed by nested PCR, which confirmed the presence of group B rotavirus in all three specimens. The sequences of the PCR products of NIV specimens were compared with that of CAL-1, ADRV and IDIR (infectious diarrhoea of infant rat) belonging to group B rotaviruses. The sequence analysis of the PCR products showed the highest identity with CAL-1, which was reported from Kolkata, India during 1997--1998. The finding suggests that human group B rotaviruses have been circulating in Pune. India, since 1993. This emerging virus may lead to more severe disease among adults in India. There is a need for surveillance of group B rotavirus infections, especially in adult diarrhoea cases and seroepidemiological studies on group B rotavirus are required among humans and animals of Western Maharashtra, India.


(Morpho)syntactic Variation in Agreement: Specificational Copular Clauses Across Germanic.
In this paper we bring together the results of our research into agreement in copular clauses in four different Germanic languages-Dutch, German, Faroese, and Icelandic-in order to provide an overview of the results. These cases present a particularly interesting window into how verbal agreement operates, since there are two potential controllers of agreement, which may disagree in person and/or number (The source of the rumor BE the neighbors/you-sg/you-pl). We will show that there is variation at all levels in which nominal controls agreement: cross-linguistic, inter-speaker within a single language, and intra-speaker. We argue that our data support the following claims: (1) "Downward" agreement for person, as well as number, with a nominal that is not in the canonical subject position is possible and in some cases preferred; (2) The agreement patterns observed in Icelandic and Faroese support the hypothesis that in these languages there are distinct Number and Person heads; (3) "Downward" agreement from a high position in the left-periphery is a grammatically distinct phenomenon from agreement when the verb remains in a lower position in the clause; (4) In some languages and some configurations, speakers show a significant degree of indeterminacy in their judgments and production, suggesting that speakers use more than one grammar. We relate our findings to current discussions in the generative literature on subject agreement and in particular differences between number and person agreement, and possible connections to restrictions on object clitics; we also discuss questions that remain open, and invite new, cross-disciplinary research.


Genetic locus half baked is necessary for morphogenesis of the ectoderm.
The zebrafish epiboly mutants partially block epiboly, the vegetalward movement of the blastoderm around the giant yolk cell. Here, we show that the epiboly mutations are located near the centromere of Linkage Group 7 in a single locus, termed the half baked locus. Nevertheless, except for the similar mutants lawine and avalanche, we find the epiboly traits of each of the alleles to be distinguishable, forming an allelic series. Using in situ analysis, we show that the specification and the formation of the germ layers is unaffected. However, during early gastrulation, convergence movements are slowed in homozygous and zygotic maternal dominant (ZMD) heterozygous mutants, especially in the epiblast layer of the blastoderm. Using triple-mutant analysis with squint and cyclops, we show that ablating involution and hypoblast formation in hab has no effect on the epiboly phenotype on the ventral and lateral sides of the embryo, suggesting that the hypoblast has no role in epiboly. Moreover, the triple mutant enhances the depletion of cells on the dorsal side of the embryo, consistent with the idea that convergence movements are defective. Double-mutant analysis with one-eyed pinhead reveals that hab is necessary in the ectodermal portion of the hatching gland. In ZMD heterozygotes, in addition to the slowing of epiboly, morphogenesis of the neural tube is abnormal, with gaps forming in the midline during segmentation stages; later, ectopic rows of neurons form in the widened spinal cord and hindbrain. Cell transplantation reveals that half baked acts both autonomously and nonautonomously in interactions among cells of the forming neural tube. Together, these results suggest that half baked is necessary within the epiblast for morphogenesis during both epiboly and neurulation and suggest that the mechanisms that drive epiboly possess common elements with those that underlie convergence and extension.


The impact of 6-month training preparation for an Ironman triathlon on the proportions of naïve, memory and senescent T cells in resting blood.
Athletes appear to be at a greater risk of illness while undertaking arduous training regimens in preparation for endurance events. As infection susceptibility has been linked with increased proportions of differentiated and senescent T cells in the periphery, changes in the proportions of these cell types due to long-term high-volume exercise training could have important implications for athlete infection risk. This study examined the effects of 6-month training preparation for an Ironman triathlon on the proportions of naïve, memory and senescent T cells in resting blood. Ten club-level triathletes (9 males; 1 female: 43 ± 3 years) were sampled at 27 (December), 21 (January), 15 (March), 9 (May) and 3 (June) weeks before an Ironman Triathlon. An additional sample was collected 2-week post-competition (August). Four-colour flow cytometry was used for the phenotypic analysis of CD4+ and CD8+ blood T cells. Proportions of differentiated (KLRG1+/CD57-) CD8+ T cells and "transitional" (CD45RA+/CD45RO+) CD4+ and CD8+ T cells increased with training, as the values in June were elevated 37, 142 and 116%, respectively, from those observed in December. Proportions of senescent (KLRG1+/CD57+) CD4+ or CD8+ T cells did not change during the training phase. Two weeks post-race, proportions of differentiated CD8+ T cells had returned to baseline values, while the proportions of senescent CD4+ T cells increased 192% alongside a 31% reduction in naïve (CD45RA+/CD45RO-) cells. In conclusion, increases in differentiated and "transitional" T cells due to arduous exercise training could compromise host protection to novel pathogens and increase athlete infection risk, although whether or not the composition of naïve and differentiated T cells in blood can serve as prognostic biomarkers in athletes remains to be established.


Central serous chorioretinopathy secondary to childbirth.
A previously healthy 37-year-old woman presented with decreased vision in the right eye, noted suddenly during childbirth. Fundus examination revealed elevation at the right macula, and optical coherence tomography confirmed central serous chorioretinopathy in the right eye. No intervention was undertaken, and the symptoms and clinical findings resolved spontaneously over 2 months.


Size-dependent oxygen activation efficiency over Pd(n)/TiO2(110) for the CO oxidation reaction.
The dissociative binding efficiency of oxygen over Pd(n)/TiO(2)(110) (n = 4, 7, 10, 20) has been measured using temperature programmed reaction (TPR) mass spectrometry and X-ray photoemission spectroscopy (XPS) following exposure to O(2) with varying doses and dose temperatures. Experiments were carried out following two different O(2) exposures at 400 K (10 L and 50 L) and for 10 L of O(2) exposure at varying temperatures (T(surf) = 200, 300, and 400 K). During TPR taken after sequential O(2) and CO (5 L at 180 K) exposures, unreacted CO is found to desorb in three features at T(desorb) ≈ 150, 200, and 430 K, while CO(2) is observed to desorb between 170 and 450 K. We show that Pd(20) has exceptionally high efficiency for oxygen activation, compared to other cluster sizes. As a consequence, its activity becomes limited by competitive CO binding at low O(2) exposures, while other Pd(n) sizes are still limited by inefficient O(2) activation. This difference in mechanism can ultimately be related back to differences in electronic properties, thus making this question one that is interesting from the theoretical perspective. We also demonstrate a correlation between one of the two CO binding sites and CO(2) production, suggesting that only CO in that site is reactive.


The role of cyclin-dependent kinases in oral potentially malignant disorders and oral squamous cell carcinoma.
Oral squamous cell carcinoma (OSCC) is a major global health problem with a relatively low-moderate 5-year survival rate. OSCC is often preceded by lesions and conditions known as oral potentially malignant disorders (OPMDs) that have an increased risk of malignant transformation. Despite advances in diagnostic technology and cancer research, the prognosis of OSCC remains poor as it is frequently detected a late stage. Understanding the molecular pathways involved in oral carcinogenesis provides a platform to identify biomarkers that may allow the early detection of OSCC and accurate prediction of the malignant potential of OPMDs. In addition, specific molecular inhibitors can be developed to target these important pathways and allow advanced therapeutic management to improve the prognosis of this malignancy. A common feature across a number of different cancers is the dysfunction of cell cycle moderator proteins known as cyclin-dependent kinases. This review summarises the current literature regarding the role of cyclin-dependent kinases in oral carcinogenesis with a particular focus on cyclin-dependent kinases 4 (CDK4) and 6 (CDK6). This is of particular relevance as CDK4 and CDK6 inhibitors have shown some promising results in other cancer types and are interesting potential treatments for OSCC.


En Route Resuscitation - Utilization of CCATT to Transport and Stabilize Critically Injured and Unstable Casualties.
The U.S. Air Force utilizes specialized Critical Care Air Transport Teams (CCATT) for transporting "stabilized" patients. Given the drawdown of military forces from various areas of operation, recent CCATT operations have increasingly involved the evacuation of unstable and incompletely resuscitated patients from far forward, austere locations. This brief report describes unique cases representative of the evolving CCATT mission and provides future direction for changes in doctrine and educational requirements in preparation for en route combat casualty care. This case series describes three patients who required significant resuscitation during CCATT transport from austere locations between April and November 2017. Approval for this project was received from the US Air Force 59th Medical Wing Institutional Review Board as non-research. Case 1: CCATT was dispatched to transport patient 1 who was reported to have a head injury after a fall. Upon evaluation of the patient onboard the aircraft, it was discovered that the patient was in cardiac arrest. Cardiopulmonary resuscitation was performed during tactical takeoff with frequent combat maneuvers. The patient developed a palpable pulse after three rounds of CPR, three doses of epinephrine, and one unit of packed red blood cells. Point of care laboratory analysis demonstrated a profoundly elevated lactate level. Cyanide poisoning was a concern but there was no antidote available in the available equipment set. After delivery to a medical facility, blood samples were positive for cyanide. Over the next 2 weeks, the patient improved and was discharged home, neurologically intact. Case 2: Patient 2 sustained complex blast injuries and bilateral lower extremity amputations. He required early transport for continuous renal replacement therapy (CRRT). The patient received 200 units of blood products in the 24 hours prior to transport and developed renal failure, pulmonary edema, and elevated ICP. During the 7 hour flight, Patient 2 received frequent adjustments of vasopressor medications, multiple Dakins solution soaks and flushes, and 1 unit of fresh frozen plasma. He remained alive 2 months later. Case 3: The team was notified to collect an urgent patient with a blast lung injury and bilateral lower extremity amputations. The ground team encountered difficulty ventilating the patient. Patient 3 arrived in the back of a pickup truck accompanied by medics and being bag valve mask ventilated with a pulse oximetry reading of 65%. He was secured to the floor of the aircraft which departed within 5 minutes of arrival. An ultrasound of the lungs showed no pneumothorax. By the end of the flight, the patient's oxygen saturation had risen to 95% and he was delivered to the emergency department in stable condition. He later passed away in the operating room due to severe blast lung and cardiac contusion. This brief report demonstrates the need of CCATT in the transport of unstable patients from forward deployed locations. The Air Force has adapted and is continuing to adapt CCATT training, equipment, onboard diagnostics and therapies, and team members' clinical skills to meet en route care combat casualty needs.


[Contact dermatoses: new antigens and current events].
Contact allergology is in perpetual evolution. The cutaneous allergens are not fixed in time but they appear and die by chance of habit or fashion changes, evolution of the chemistry, the cosmetology or the allergologist campaign. It seems interesting to consider the recent data concerning classical or new allergens.


[Intra-abdominal hypertension in patients with severe acute pancreatitis].
The role of parapancreatitis in the intra-abdominal hypertension (IAH) progression in patients with severe acute pancreatitis with various forms of organs failure was studied. It was analyzed the treatment results of 63 patients with severe acute pancreatitis. The patients were divided into 4 groups: the first group (n=24) had not the signs of organs failure, the second group (n=15) - with symptoms of acute respiratory failure (ARF), the third group (n=11) - with a combination of ARF and acute cardiovascular failure (CVF) and the fourth group (n=13) - with a combination of ARF, CVF and acute renal failure. Intra-abdominal pressure (IAP) was measured at 1, 3, 5, 7- days from the beginning of the disease with calculation of the abdominal perfusion pressure and filtration gradient values. The prevalence evaluation of parapancreatitis was carried out on the basis of CT-data with bolus-dosing of the contrast agent, ultrasound, video laparoscopy as well as data obtained during operation and autopsy. It was defined that the intra-abdominal pressure (IAP) values was not significantly different and complied with intra-abdominal hypertension of the first grade in patients of the groups 1-3 at the first day. IAP was significantly higher and complied with intra-abdominal hypertension of the second grade in patients of the fourth group. IAP normalized to 5-7th days in patients of the first and the second groups. The patients of the third and the fourth groups had IAH of the second grade in the 7th day. There was reliable (p<0.01) average positive correlation (r=0.57) between the indications of IAH and the scale APACHE II. The patients with common parapancreatitis (n=39) had indications of IAP and APACHE II significantly higher than in patients with local forms of parapancreatitis (n=24; p<0.01). Common defeat of retroperitoneal fat determined persistent increase of IAP more than the presence of effusion in the abdominal cavity. In case of the first grade of IAH the mortality was 6.6%, the second grade - 37.5%, the third grade - 58.3% and the fourth grade - 80%. All died patients had a common parapancreatitis. The authors consider that the indications of IAP and APACHE II let to suppose an adverse outcome by the development of multiple organ failure in the early stages of the disease. Common parapancreatitis is the main feature of the persistent IAH (IAP increase during 5-7 days), which is an indication for early decompression operations on the abdominal wall.


The control of saccharide synthesis during development of Myxamoebae of Dictyostelium discoideum containing differing amounts of glycogen.
1. Myxamoebae initially containing 5.59mg of glycogen/10(8) cells accumulate approx. 25% more cell-wall polysaccharide, 100% more mucopolysaccharide, 200% more glucose and 300% more trehalose during their development than do myxamoebae initially containing less than 0.3mg of glycogen/10(8) cells. 2. These observations restrict the number of possible control mechanisms operating to regulate carbohydrate metabolism during development. 3. Cells accumulating a large amount of trehalose (approx. 400mug/10(8) cells) have the same amount and pattern of changes in specific activity of trehalase and trehalose 6-phosphate synthase as do cells accumulating a smaller amount of trehalose (approx. 100mug/10(8) cells). 4. These two populations of cells do, however, differ markedly in the amount of UDP-glucose and glucose 6-phosphate that they contain. 5. It is concluded that this change in the intracellular pools of the metabolic precursors of trehalose accounts for the increased amount of trehalose synthesized by cells derived from myxamoebae containing an increased glycogen content.


In vitro germ cell models for the detection of fertility impairment.
Pluripotent embryonic carcinoma cells and pluripotent embryonic stem cells established from undifferentiated cells of an early mouse embryo were investigated for induction of proliferation inhibition, sister chromatid exchanges (SCE) and single-strand breaks by treatment with various germ cell mutagens. The comparison of malignant cells with nonmalignant cells showed an increased sensitivity of nonmalignant cells independent of their state of differentiation. Mitomycin C (MMC) inhibited the proliferation of nonmalignant cells at a concentration of 10(-6) M but did not affect growth of the teratocarcinoma cell line P19. There were no differences between the investigated cell lines at a lower MMC concentration. At the concentration of 10(-6) M MMC the sister chromatid exchanges of P19 were enhanced up to 41 SCE per metaphase. Testing of another germ cell mutagen, ethylnitrosourea (ENU), gave similar results: a decreasing generation time of nonmalignant cell lines after treatment with 1 mM ENU and no effect on the teratocarcinoma cells. This concentration also induced a high number of SCE. Single-strand breaks could be produced by exposure to methanmethylsulphonate (MMS). 56.3% of embryonic stem cell DNA was passing through the filter after MMS treatment. In contrast to the embryonic stem cells, only 35.6% of teratocarcinoma DNA was affected.


Starch fermentation by recombinant saccharomyces cerevisiae strains expressing the alpha-amylase and glucoamylase genes from lipomyces kononenkoae and saccharomycopsis fibuligera.
Lipomyces kononenkoae and Saccharomycopsis fibuligera possess highly efficient alpha-amylase and/or glucoamylase activities that enable both of these yeasts to utilize raw starch as a carbon source. Eight constructs containing the L. kononenkoae alpha-amylase genes (LKA1 and LKA2), and the S. fibuligera alpha-amylase (SFA1) and glucoamylase (SFG1) genes were prepared. The first set of constructs comprised four single gene cassettes each containing one of the individual amylase coding sequences (LKA1, LKA2, SFA1 or SFG1) under the control of the phosphoglycerate kinase gene (PGK1) promoter and terminator, while the second set comprised two single cassettes containing SFA1 and SFG1 linked to their respective native promoters and terminators. The third set of constructs consisted of two double-gene cassettes, one containing LKA1 plus LKA2 under the control of the PGK1 promoter and terminator, and the other SFA1 plus SFG1 controlled by their respective native promoters and terminators. These constructs were transformed into a laboratory strain Saccharomyces cerevisiae (Sigma1278b). Southern-blot analysis confirmed the stable integration of the different gene constructs into the S. cerevisiae genome and plate assays revealed amylolytic activity. The strain expressing LKA1 and LKA2 resulted in the highest levels of alpha-amylase activity in liquid media. This strain was also the most efficient at starch utilization in batch fermentations, utilizing 80% of the available starch and producing 0.61g/100 mL of ethanol after 6 days of fermentation. The strain expressing SFG1 under the control of the PGK1 expression cassette gave the highest levels of glucoamylase activity. It was shown that the co-expression of these heterologous alpha-amylase and glucoamylase genes enhance starch degradation additively in S. cerevisiae. This study has resulted in progress towards laying the foundation for the possible development of efficient starch-degrading S. cerevisiae strains that could eventually be used in consolidated bioprocessing, and in the brewing, whisky, and biofuel industries.


The emergence of mesolithic cemeteries in SW Europe: insights from the El Collado (Oliva, Valencia, Spain) radiocarbon record.
Located on the Iberian Mediterranean coast, El Collado is an open-air site where a rescue excavation was conducted over two seasons in 1987 and 1988. The archaeological work excavated a surface area of 143 m2 where 14 burials were discovered, providing skeletal remains from 15 individuals. We have obtained AMS dates for 10 of the 15 individuals by means of the direct dating of human bones. The ranges of the probability distribution of the calibrated dates suggest that the cemetery was used during a long period of time (781-1020 years at a probability of 95.4%). The new dates consequently set back the chrono-cultural attribution of the cemetery from the initial proposal of Late Mesolithic to an older date in the Early Mesolithic. Therefore, El Collado becomes the oldest known cemetery in the Iberian Peninsula, earlier than the numerous Mesolithic funerary contexts documented on the Atlantic façade such as the Portuguese shell-middens in the Muge and Sado Estuaries or the funerary sites on the northern Iberian coast.


Usefulness of serum procalcitonin level for prediction of vesicoureteral reflux in pediatric urinary tract infection.
Procalcitonin is a sensitive biomarker for bacterial infections. Recent studies show a correlation between serum procalcitonin level and vesicoureteral reflux (VUR). The aim of this study was to evaluate the predictive value of procalcitonin in diagnosis of VUR in children with febrile urinary tract infection. One hundred and eight children aged 2 month to 12 years with febrile urinary tract infection were evaluated. Serum procalcitonin was measured before initiation of antibiotics. Standard voiding cystourethrography (VCUG) was performed in all children as the gold standard for detection of VUR. Sensitivity and specificity of a high procalcitonin level was evaluated using the receiver operating characteristic curve. Forty-eight patients (44%) had VUR at least in one kidney, including grade 1 to 2 in 12 patients (11.1%), grade 3 in 16 (14.8%), and grade 4 to 5 in 20 patients (18.5%). Procalcitonin level ranged from 0.05 ng/mL to 13.6 ng/mL. Procalcitonin level was significantly higher with increasing the grading of reflux. Comparing procalcitonin levels with VCUG results, a sensitivity of 97% and a specificity of 75% was obtained at a procalcitonin level of 0.59 ng/mL for diagnosis of VUR. There was a significant correlation between procalcitonin level and leukocytosis, erythrocyte sedimentation rate, and C-reactive protein. A high procalcitonin level may be used for prediction of all grades of VUR in children with febrile urinary tract infection. A low procalcitonin level may be used for avoidance of unnecessary VCUG in some low-risk patients.


Reference values of pulse wave velocity in healthy children and teenagers.
Carotid-femoral pulse wave velocity is an established method for characterizing aortic stiffness, an individual predictor of cardiovascular mortality in adults. Normal pulse wave velocity values for the pediatric population derived from a large data collection have yet to be available. The aim of this study was to create a reference database and to characterize the factors determining pulse wave velocity in children and teenagers. Carotid-femoral pulse wave velocity was measured by applanation tonometry. Reference tables from pulse wave velocities obtained in 1008 healthy subjects (aged between 6 and 20 years; 495 males) were generated using a maximum-likelihood curve-fitting technique for calculating SD scores in accordance with the skewed distribution of the raw data. Effects of sex, age, height, weight, blood pressure, and heart rate on pulse wave velocity were assessed. Sex-specific reference tables and curves for age and height are presented. Pulse wave velocity correlated positively (P<0.001) with age, height, weight, and blood pressure while correlating negatively with heart rate. After multiple regression analysis, age, height, and blood pressure remained major predictors of pulse wave velocity. This study, involving >1000 children, is the first to provide reference values for pulse wave velocity in children and teenagers, thereby constituting a suitable tool for longitudinal clinical studies assessing subgroups of children who are at long-term risk of cardiovascular disease.