Effective Number Theory: Counting the Identities of a Quantum State.
Quantum physics frequently involves a need to count the states, subspaces, measurement outcomes, and other elements of quantum dynamics. However, with quantum mechanics assigning probabilities to such objects, it is often desirable to work with the notion of a "total" that takes into account their varied relevance. For example, such an effective count of position states available to a lattice electron could characterize its localization properties. Similarly, the effective total of outcomes in the measurement step of a quantum computation relates to the efficiency of the quantum algorithm. Despite a broad need for effective counting, a well-founded prescription has not been formulated. Instead, the assignments that do not respect the measure-like nature of the concept, such as versions of the participation number or exponentiated entropies, are used in some areas. Here, we develop the additive theory of effective number functions (ENFs), namely functions assigning consistent totals to collections of objects endowed with probability weights. Our analysis reveals the existence of a minimal total, realized by the unique ENF, which leads to effective counting with absolute meaning. Touching upon the nature of the measure, our results may find applications not only in quantum physics, but also in other quantitative sciences.


[Comparative study of normal and over-users of a health center].
To compare normal users (NU) and over-users (OU), examining social and demographic features, family characteristics, presence of psychological problems or family dysfunction and the existence of chronic pathologies. An observational crossover study, lasting one year (1994). Health centre. Randomised sampling of 12,092 patients > 14 years old. OU (n = 260) was defined as someone who had a percentage point for attendance without an appointment at the Centre, excluding bureaucratic reasons, of above 75%; and NU (n = 260), in the 25 and 75% band. 65 individuals (240 OU and 225 NU) took part. Average age of OU was higher than that of NU. 88 OU (36.7%) finished their education above primary level, whereas 115 NU (51.1%) did so (OR 1.81). 37.5% OU (90) belonged to small families, against 24.9% NU (56). 34.6% OU (83) were in an advanced family vital cycle, against 24% NU (54. 26.7% OU (64) and 17.8% NU (40) had psychological problems. There was family dysfunction in 32 OU (13.3%) and 23% NU (10.2%). 186 OU (77.5%) had some chronic pathology, against 96 NU, of whom 70.3% OU and 87.2% NU attended appointments arranged to monitor these pathologies. Over-use of our health centre is related to being older, a low level of education, belonging to small families in an advanced vital cycle, having psychological problems, suffering chronic pathologies and not attending appointments when they are arranged.


The CCAAT/enhancer-binding protein-ATF response elements-luciferase mouse model, an innovative tool to monitor the integrated stress response pathway in vivo.
The article highlights the recent development of an ATF4 (activating transcription factor) inducible luciferase (LUC) mouse model to monitor the integrated stress response pathway (ISR) in vivo. The ISR pathway plays a key role in cellular adaptation to stress and is dysregulated in numerous diseases. The core event in this pathway is the phosphorylation of eukaryotic translation initiation factor 2 α, which leads to the recruitment of the transcription factor ATF4 to specific CCAAT/enhancer-binding protein-ATF response elements (CAREs) located in the promoters of target genes. To monitor the modulation of this pathway in the whole animal and at tissue and cellular levels, we generated a CARE-driven LUC mouse model. We validated the relevance of this model to study stress-related pathologies and recently observed the correlation between the ISR pathway induction in muscle and the occurrence of stress-induced skeletal muscle atrophy. The CARE-LUC mouse model represents an innovative tool for investigating the role of the ISR pathway in physiology and disease and opens new avenues for the development of drugs that could modify this important pathway in stress-related human diseases.


A cognitively enhanced online Tai Ji Quan training intervention for community-dwelling older adults with mild cognitive impairment: A feasibility trial.
This study examines the feasibility, acceptability, and safety of a newly developed cognitive-enhancing Tai Ji Quan training intervention, delivered via remote videoconferencing, for older adults with mild cognitive impairment (MCI). In a three-arm feasibility trial, community-dwelling older adults with MCI (N = 69; mean age = 74.6 years, 57% women) were randomized to a cognitively enhanced Tai Ji Quan (n = 23), standard Tai Ji Quan (n = 22), or stretching group (n = 24) and participated in a 60-minute online exercise session via Zoom, twice weekly for 16 weeks. Participants were recruited primarily in the state of Oregon through mass mailing and word of mouth. The primary outcomes were intervention feasibility (with respect to recruitment, online intervention delivery, fidelity and compliance, and attrition and retention rates), acceptability, and safety. We also assessed feasibility of online data collection and test-retest reliability and explored preliminary trends on secondary outcomes that included global cognitive function, dual-task cost, and domain-specific cognition function. The study had an average recruitment rate of 55%. Feasibility was demonstrated by the overall successful online program implementation, with good fidelity, acceptable compliance (76%), and excellent retention (94%). The cognitively enhanced Tai Ji Quan intervention was shown to be acceptable to participants as well as safe, with no major intervention-related moderate/severe events. At week 16, the group receiving cognitively enhanced Tai Ji Quan training showed a positive trend in the cognitive function and dual-task outcome measures whereas the group receiving standard Tai Ji Quan training exhibited positive trends on global and domain-specific cognitive measures. Preliminary findings of this pilot study indicate the feasibility, acceptability, and safety of a tailored, cognitively enhanced Tai Ji Quan training intervention delivered remotely to home settings via videoconferencing for community-dwelling older adults with MCI. Clinicaltrials.gov identifier NCT04070703.


Association between Responsible Pet Ownership and Glycemic Control in Youths with Type 1 Diabetes.
Type 1 diabetes mellitus (T1DM) a chronic characterized by an absolute insulin deficiency requires conscientious patient self-management to maintain glucose control within a normal range. Family cohesion and adaptability, positive coping strategies, social support and adequate self-regulatory behavior are found to favorably influence glycemic control. Our hypothesis was that the responsible care of a companion animal is associated with these positive attributes and correlated with the successful management of a chronic illness such as type 1 diabetes. We recruited 223 youths between 9 and 19 years of age from the Pediatric Diabetes clinic at the University of Massachusetts Medical School, reviewed the status of their glycemic control (using three consecutive A1c values) and asked them questions about the presence of a pet at home, and their level of involvement with its care. Multivariate analyses show that children who care actively for one or more pets at home are 2.5 times more likely to have control over their glycemic levels than children who do not care for a pet, adjusting for duration of disease, socio-economic status, age and self-management [1.1 to 5.8], pWald = 0.032. A separate model involving the care of a petdog only yielded comparable results (ORa = 2.6 [1.1 to 5.9], pWald = 0.023).


The fate of drugs in pregnancy.
The response of mother and fetus/neonate to drugs administered to the pregnant woman is determined largely by drug disposition and elimination within and between mother and fetus. Physiological changes occurring in pregnancy may result in reduced plasma protein binding, an increase in the apparent volume of distribution, and more rapid metabolic and renal clearance of certain drugs than are usually found in non-pregnant women. The consequences are relevant for the peak concentration of drug achieved after a single dose, the half-life of the drug, and the drug concentration to which the fetus is exposed via the placenta. Drugs whose dosage may need to be altered in pregnancy include most anticonvulsants, lithium, digoxin, certain beta-blockers, ampicillin and cefuroxime; for drugs which have not been specifically investigated, no generalizations are possible. Very few drugs given in pregnancy fail to cross the placenta and, as a rule, drug molecules not bound to plasma protein diffuse along a concentration gradient to establish and maintain an equilibrium. Because movement of drug molecules is bidirectional, the placenta is the main portal of exit from, as well as entry to, the fetus. The pharmacological response of the fetus to drugs which it receives depends mainly on the unbound concentration of drug in fetal blood--unwanted effects seldom occur if the maternal dose is correct; exceptions include tetracycline, antithyroid drugs, coumarin anticoagulants, aspirin, indomethacin and cerebral depressant drugs (opiates, barbiturates and phenothiazines). The total (bound plus unbound) drug concentration on the fetal side of the placenta is usually lower than on the maternal side except where differences in blood pH and/or protein binding lead to 'trapping' of drug in the fetus; the latter phenomenon may prove hazardous after birth because of the neonate's limited capacity to eliminate drugs by metabolism and excretion.


Paclitaxel associated with cholesterol-rich nanoemulsions promotes atherosclerosis regression in the rabbit.
A cholesterol-rich nanoemulsion (LDE) that resembles LDL binds to the LDL receptors and after injection into the blood stream may concentrate in cells with LDL receptor overexpression, as occurs in neoplasias and other proliferative processes. Thus, LDE can be used as vehicle to target drugs against those cells. The current study was designed to verify in rabbits whether LDE concentrates in the lesioned rabbit artery and whether a paclitaxel derivative, paclitaxel oleate, associated to LDE could reduce the atherosclerotic lesions. Sixteen male New Zealand rabbits were fed a 1% cholesterol diet for 60 days. Starting from day 30 under cholesterol feeding, eight animals were treated with four weekly intravenous injections of LDE-paclitaxel (4 mg/kg) and eight with four weekly intravenous saline solution injections for additional 30 days. On day 60, the animals were sacrificed for analysis. The uptake of LDE labeled with [(14)C]-cholesteryl oleate by the aortic arch of cholesterol-fed rabbits was twice as much that observed in animals fed only regular chow. LDE-paclitaxel reduced the lesion areas of cholesterol-fed animals by 60% and intima-media ratio fourfold and inhibited the macrophage migration and the smooth muscle cell proliferation and invasion of the intima. LDE-paclitaxel treatment had no toxicity. In conclusion, LDE-paclitaxel produced pronounced atherosclerosis regression without toxicity and has shown remarkable potential in cardiovascular therapeutics.


Pancreatic progenitor epigenome maps prioritize type 2 diabetes risk genes with roles in development.
Genetic variants associated with type 2 diabetes (T2D) risk affect gene regulation in metabolically relevant tissues, such as pancreatic islets. Here, we investigated contributions of regulatory programs active during pancreatic development to T2D risk. Generation of chromatin maps from developmental precursors throughout pancreatic differentiation of human embryonic stem cells (hESCs) identifies enrichment of T2D variants in pancreatic progenitor-specific stretch enhancers that are not active in islets. Genes associated with progenitor-specific stretch enhancers are predicted to regulate developmental processes, most notably tissue morphogenesis. Through gene editing in hESCs, we demonstrate that progenitor-specific enhancers harboring T2D-associated variants regulate cell polarity genes LAMA1 and CRB2. Knockdown of lama1 or crb2 in zebrafish embryos causes a defect in pancreas morphogenesis and impairs islet cell development. Together, our findings reveal that a subset of T2D risk variants specifically affects pancreatic developmental programs, suggesting that dysregulation of developmental processes can predispose to T2D.


Cleft lip with or without cleft palate: implication of the heavy chain of non-muscle myosin IIA.
Non-syndromic cleft lip with or without palate (CL/P) is one of the most common malformations among live births, but most of the genetic components and environmental factors involved remain to be identified. Among the different causes, MYH9, the gene encoding for the heavy chain of non-muscle myosin IIA, was considered a potential candidate, because it was found to be abundantly and specifically expressed in epithelial cells of palatal shelves before fusion. After fusion, its expression level was shown to decrease and to become limited to epithelial triangles before disappearing, as fusion is completed. To determine whether MYH9 plays a role in CL/P aetiology, a family-based association analysis was performed in 218 case/parent triads using single-nucleotide polymorphism (SNP) markers. Pairwise and multilocus haplotype analyses identified linkage disequilibrium between polymorphism alleles at the MYH9 locus and the disease. The strongest deviation from a null hypothesis of random sharing was obtained with two adjacent SNPs, rs3752462 and rs2009930 (global p value = 0.001), indicating that MYH9 might be a predisposing factor for CL/P, although its pathogenetic role needs to be investigated more accurately.


Total costs and atrial fibrillation ablation success or failure in Medicare-aged patients in the United States.
This retrospective cohort study compared the direct medical costs of successful versus unsuccessful catheter ablation in Medicare-aged patients with atrial fibrillation (AF), using medical claims data. AF patients with > or = 12 months of continuous medical/pharmacy coverage pre- and postablation were identified from the MarketScan Medicare database (January 2003 to December 2006). For study inclusion, patients were required to have > or = 2 AF inpatient/outpatient visits within 6 months and to have received antiarrhythmic drug therapy within 12 months prior to the index ablation. Ablation success was defined as the absence of antiarrhythmic drug therapy 6-12 months postablation. Of 135 patients identified (67% men, mean age 73 years), ablation was successful in 69 (51.1%); most patients (96%) underwent a single procedure. Patients with successful ablation discontinued antiarrhythmic drug treatment after (mean) 54 days. Use of rate-control and anticoagulant drugs decreased after successful ablation, from 87% to 67% and from 86% to 64% of patients, respectively. Among failed ablation patients, 74% versus 70% received rate-control drugs, and 88% versus 82% received anticoagulants pre- versus postablation. Mean +/- SD per-patient procedural costs were $13,655+/-$12,761 for successful compared with $17,294+/-$26,502 (P=0.21) for failed ablation, while AF-related medical costs over 12 months postablation were $2394+/-$642 and $2703+/-$1706, respectively (P<0.001). Overall costs tended to be lower for successful ($16,049+/-$12,536) than for failed ($19,997+/-$13,958) AF ablation (P=0.07). These findings are subject to the limitations imposed by a retrospective database analysis and a small sample size. Outside the clinical-trial setting, catheter ablation for second-line treatment of AF proved unsuccessful in half of Medicare-aged patients. Direct medical costs did not differ significantly between patients with failed and successful ablations. The high rate and costs of AF ablation failure in the Medicare-aged population reinforce the need for better understanding of prognostic factors for ablation outcome.


Gallbladder cancer: surgical management.
Gallbladder cancer is the most common biliary tract malignancy. Margin-clearing surgery is a cornerstone of gallbladder cancer management, but several aspects of surgical management are controversial. This review will discuss the current state of surgical management for gallbladder cancer as well as aspects of gallbladder surgery that remain debated, including operative extent, lymphadenectomy extent, and management of incidentally discovered gallbladder cancer.


Ethnic differences in functional and neuropsychological test performance in older adults.
Numerous studies outline discrepancies in neuropsychological test profiles in African American and European American samples, despite similarities on major background factors. In our clinical sample of convenience, African American and European American older adults did not diverge on age, years of formal education, or global cognitive impairment. We predicted that ethnic groups would differ on a financial index, reading, and naming measures, but not on form discrimination and health problem-solving tasks. Results showed the expected patterns with the exception of form discrimination, which was lower in the African American group. Ethnicity no longer predicted standing on financial and nonverbal discrimination measures once we adjusted scores to equate for word-reading ability. In contrast, ethnicity status continued to explain significant variance in confrontation naming following adjustment for reading. When ethnic groups were subdivided by sex, European American men tended to have the highest scores of all four subgroups despite having equivalent levels of education. Therefore, future studies should examine the impact of cognitive activity throughout adulthood on real-world function and neuropsychological test performance among groups defined by both ethnicity and sex. In conclusion, in African American and European American elders seen for clinical assessment, literacy appeared to be associated with discrepant scores on a measure of real-world financial knowledge and skill, as well as with differences in verbal and nonverbal neuropsychological measures.


[An ischemic syndrome of the oculumotor nucleus: associated clinical and anatomical variations on a theme].
Nuclear syndrome of the oculomotor nerve was first described in 1981, it is characterized by the association of an ipsilateral third nerve palsy with a paresis of elevation in the contralateral eye. This syndrome can be caused by vascular or tumoral lesions in the upper midbrain. It is rarely due to ischemic unilateral mesencephalic lesions, because ischemic lesions of the midbrain are usually integrated in a diffuse involvement of the brainstem and the thalamo-sub-thalamic region. In case of nuclear syndrome of the third nerve due to isolated upper midbrain infarct in the paramedian territory, dependent on branches of the basilar artery, oculomotor symptoms are frequently isolated. On the contrary, in fascicular syndromes of the third nerve, resulting from stroke in more lateral territories upon branches of the posterior cerebral artery, many neurological symptoms are associated with the oculomotor signs. We describe 3 patients presenting with a characteristic nuclear syndrome of the third nerve, resulting from a unilateral paramedian ischemic stroke in the upper midbrain, confirmed by cerebral CT scan or MRI examination. Clinical presentation differed in each case, and marked contralateral hemiparesia, cerebellar syndrome and focal asterixis were associated in various ways with the stereotyped oculomotor disorders. In the 3 cases, the nuclear syndrome of the third nerve was associated with fascicular involvement of the nerve, in an unusual clinical picture. The theoretical distinction between nuclear and fascicular syndromes is supported by the anatomical description of the arterial segmentation in the upper midbrain, which remains debated since the first description. According to the variability of clinical presentations, it seems that the arterial territories may be more variable than initially described. Therefore, ischemic lesions of the upper midbrain may involve some vascular borderzones with a high inter-individual variability. Upper midbrain strokes may thus lead to variable clinical pictures.


CRISPR/Cas9-based depletion of 16S ribosomal RNA improves library complexity of single-cell RNA-sequencing.
Single-cell RNA-sequencing (scRNA-seq) relies on PCR amplification to retrieve information from vanishingly small amounts of starting material. To selectively enrich mRNA from abundant non-polyadenylated transcripts, poly(A) selection is a key step during library preparation. However, some transcripts, such as mitochondrial genes, can escape this elimination and overwhelm libraries. Often, these transcripts are removed in silico, but whether physical depletion improves detection of rare transcripts in single cells is unclear. We find that a single 16S ribosomal RNA is widely enriched in planarian scRNA-seq datasets, independent of the library preparation method. To deplete this transcript from scRNA-seq libraries, we design 30 single-guide RNAs spanning its length. To evaluate the effects of depletion, we perform a side-by-side comparison of the effects of eliminating the 16S transcript and find a substantial increase in the number of genes detected per cell, coupled with virtually complete loss of the 16S RNA. Moreover, we systematically determine that library complexity increases with a limited number of PCR cycles following CRISPR treatment. When compared to in silico depletion of 16S, physically removing it reduces dropout rates, retrieves more clusters, and reveals more differentially-expressed genes. Our results show that abundant transcripts reduce the retrieval of informative transcripts in scRNA-seq and distort the analysis. Physical removal of these contaminants enables the detection of rare transcripts at lower sequencing depth, and also outperforms in silico depletion. Importantly, this method can be easily customized to deplete any abundant transcript from scRNA-seq libraries.


Terahertz Spectroscopy for Proximal Soil Sensing: An Approach to Particle Size Analysis.
Spatially resolved soil parameters are some of the most important pieces of information for precision agriculture. These parameters, especially the particle size distribution (texture), are costly to measure by conventional laboratory methods, and thus, in situ assessment has become the focus of a new discipline called proximal soil sensing. Terahertz (THz) radiation is a promising method for nondestructive in situ measurements. The THz frequency range from 258 gigahertz (GHz) to 350 GHz provides a good compromise between soil penetration and the interaction of the electromagnetic waves with soil compounds. In particular, soil physical parameters influence THz measurements. This paper presents investigations of the spectral transmission signals from samples of different particle size fractions relevant for soil characterization. The sample thickness ranged from 5 to 17 mm. The transmission of THz waves was affected by the main mineral particle fractions, sand, silt and clay. The resulting signal changes systematically according to particle sizes larger than half the wavelength. It can be concluded that THz spectroscopic measurements provide information about soil texture and penetrate samples with thicknesses in the cm range.


A monatomic system with a liquid-liquid critical point and two distinct glassy states.
We study the glass transition (GT) in a model system that exhibits the presence of more than one liquid or glassy state ("polyamorphism") using molecular dynamics simulations. We study the Jagla model [E. A. Jagla, J. Chem. Phys. 111, 8980 (1999)], a two-scale spherically symmetric ramp potential with both attractive and repulsive interactions. The Jagla model is particularly interesting since, depending on its parametrization, it predicts two phases ("polyamorphism") not only in the glassy state but also in equilibrium as a liquid-liquid phase transition (LLPT). The Jagla model may also be useful in understanding a recent observation of polyamorphism in metallic glasses containing cerium. We use a parametrization for which crystallization can be avoided and the GT and LLPT are clearly separated, providing a unique opportunity to study the effects of the LLPT on the GT. We follow the experimental protocol employed in the classical differential scanning calorimetry experiments used to characterize the GT, cooling and heating the system through the GT and calculating the constant-pressure specific heat C(P) and the thermal expansion coefficient alpha(P). At pressures below and well above the LLPT, the same basic GT phenomenology of metallic glasses is observed, i.e., a single peak in C(P) (typical of ergodicity restoration) occurs upon heating across the GT. At pressures above the LLPT, a second peak in C(P) develops at higher temperature above the GT. This second peak in C(P) arises from the presence of a Widom line T(W) defined as the locus of maximum correlation length in the one-phase region above the liquid-liquid critical point (LLCP). The behavior of alpha(P) is different across the GT and Widom line. Near the GT temperature T(g), alpha(P) displays a small peak upon heating, which makes a negligible contribution to the C(P) peak. On the other hand, near T(W), alpha(P) displays a much larger peak, which makes a substantial contribution to the C(P) peak at higher temperature. We find that T(g) is almost independent of pressure for each of the two coexisting liquids, but shows an apparent discontinuity upon crossing the LLPT line, to a lower value for the higher-entropy phase. We compare the entropies of both phases, and the corresponding temperature dependencies, with those of the crystal phase. We also study the dependence of the GT on heating rate and find that for pressures below the LLCP, slow heating results in crystallization, as occurs in laboratory experiments. Regarding the thermal expansion properties of the Jagla model, we study the interplay of the density minimum recently observed in confined water and the GT.


Competition among Nasal Bacteria Suggests a Role for Siderophore-Mediated Interactions in Shaping the Human Nasal Microbiota.
Resources available in the human nasal cavity are limited. Therefore, to successfully colonize the nasal cavity, bacteria must compete for scarce nutrients. Competition may occur directly through interference (e.g., antibiotics) or indirectly by nutrient sequestration. To investigate the nature of nasal bacterial competition, we performed coculture inhibition assays between nasal Actinobacteria and Staphylococcus spp. We found that isolates of coagulase-negative staphylococci (CoNS) were sensitive to growth inhibition by Actinobacteria but that Staphylococcus aureus isolates were resistant to inhibition. Among Actinobacteria, we observed that Corynebacterium spp. were variable in their ability to inhibit CoNS. We sequenced the genomes of 10 Corynebacterium species isolates, including 3 Corynebacterium propinquum isolates that strongly inhibited CoNS and 7 other Corynebacterium species isolates that only weakly inhibited CoNS. Using a comparative genomics approach, we found that the C. propinquum genomes were enriched in genes for iron acquisition and harbored a biosynthetic gene cluster (BGC) for siderophore production, absent in the noninhibitory Corynebacterium species genomes. Using a chrome azurol S assay, we confirmed that C. propinquum produced siderophores. We demonstrated that iron supplementation rescued CoNS from inhibition by C. propinquum, suggesting that inhibition was due to iron restriction through siderophore production. Through comparative metabolomics and molecular networking, we identified the siderophore produced by C. propinquum as dehydroxynocardamine. Finally, we confirmed that the dehydroxynocardamine BGC is expressed in vivo by analyzing human nasal metatranscriptomes from the NIH Human Microbiome Project. Together, our results suggest that bacteria produce siderophores to compete for limited available iron in the nasal cavity and improve their fitness.IMPORTANCE Within the nasal cavity, interference competition through antimicrobial production is prevalent. For instance, nasal Staphylococcus species strains can inhibit the growth of other bacteria through the production of nonribosomal peptides and ribosomally synthesized and posttranslationally modified peptides. In contrast, bacteria engaging in exploitation competition modify the external environment to prevent competitors from growing, usually by hindering access to or depleting essential nutrients. As the nasal cavity is a nutrient-limited environment, we hypothesized that exploitation competition occurs in this system. We determined that Corynebacterium propinquum produces an iron-chelating siderophore, and this iron-sequestering molecule correlates with the ability to inhibit the growth of coagulase-negative staphylococci. Furthermore, we found that the genes required for siderophore production are expressed in vivo Thus, although siderophore production by bacteria is often considered a virulence trait, our work indicates that bacteria may produce siderophores to compete for limited iron in the human nasal cavity.


Antigen 43 from Escherichia coli induces inter- and intraspecies cell aggregation and changes in colony morphology of Pseudomonas fluorescens.
Antigen 43 (Ag43) is a surface-displayed autotransporter protein of Escherichia coli. By virtue of its self-association characteristics, this protein is able to mediate autoaggregation and flocculation of E. coli cells in static cultures. Additionally, surface display of Ag43 is associated with a distinct frizzy colony morphology in E. coli. Here we show that Ag43 can be expressed in a functional form on the surface of the environmentally important Pseudomonas fluorescens strain SBW25 with ensuing cell aggregation and frizzy colony types. Using green fluorescence protein-tagged cells, we demonstrate that Ag43 can be used as a tool to provide interspecies cell aggregation between E. coli and P. fluorescens. Furthermore, Ag43 expression enhances biofilm formation in P. fluorescens to glass surfaces. The versatility of this protein was also reflected in Ag43 surface display in a variety of other gram-negative bacteria. Display of heterologous Ag43 in selected bacteria might offer opportunities for rational design of multispecies consortia where the concerted action of several bacterial species is required, e.g., waste treatment and degradation of pollutants.


Essentials of slit lamp biomicroscopy.
Biomicroscopy is an indispensable part of the ocular health examination performed on every patient. The purpose of this paper is to briefly review the basic illumination technique so that the examiner thoroughly understands their uses and develops for himself a specific investigative procedure. This testing may progress from the lids and lashes to the anterior vitreous in a coordinated manner.


A statistical evaluation of the potential genotoxic activity in the surface waters of the Golden Horn Estuary.
The potential genotoxic activity in the surface waters of the Golden Horn Estuary was statistically evaluated utilizing a combination of appropriate parametric and non-parametric tests. The genotoxic activities that were associated with the water samples were determined by the SOS chromotest microplate assay. This assay utilizes β-galactosidase activity, alkaline phosphatase activity, and four different solvent controls, to generate reliable results when corrected induction factors (CIF) are used as quantitative measurements of genotoxic activity. The CIF values were obtained from a total of 384 different genotoxic experiments that were grouped into subsets according to the respective seasons and the selected sampling locations. A total of 160 subsets were statistically compared to assess any possible differences between the pairs of groups, with 95% confidence limits. The findings of this study clearly indicate that some seasonal variations exist in the CIF values at several sampling sites. However, no potentially hazardous impact to the aquatic environment was found in the estuarine system.


Anterior Cervical Reconstruction Using Free Vascularized Fibular Graft after Cervical Corpectomy.
Study Design Prospective study. Objective The aim of this study was to evaluate the clinical and radiologic results of using free vascularized fibular graft (FVFG) for anterior reconstruction of the cervical spine following with varying levels of corpectomy. Methods Ten patients underwent anterior cervical reconstruction using an FVFG after cervical corpectomy augmented with internal instrumentation. All patients were evaluated neurologically according to the Japanese Orthopaedic Association (JOA) and modified JOA scoring systems and the Nurick grading system. The neurologic recovery rate was determined, and the clinical outcome was assessed based on three factors: neck pain, dependence on pain medication, and ability to return to work. The fusion status and maintenance of lordotic correction by the strut graft were determined by measuring the lordosis angle and fused segment height (FSH). Results All patients achieved successful fusion. The mean follow-up period was 35.2 months (range, 28 to 44 months). Graft union occurred at a mean of 3.5 months. The mean loss of lordotic correction was 0.95 degrees, and the mean change in FSH was <1 mm. The neurologic recovery rate was excellent in four patients, good in five, and fair in one. All patients achieved satisfactory clinical outcome. No neurologic injuries occurred during the operations. Conclusion The use of FVFG is a valuable and effective technique in anterior cervical reconstruction for complex disorders.


IL-10 is necessary for the expression of airway hyperresponsiveness but not pulmonary inflammation after allergic sensitization.
Cytokines play an important role in modulating inflammatory responses and, as a result, airway tone. IL-10 is a regulatory cytokine that has been suggested for treatment of asthma because of its immunosuppressive and anti-inflammatory properties. In contrast to these suggestions, we demonstrate in a model of allergic sensitization that mice deficient in IL-10 (IL-10-/-) develop a pulmonary inflammatory response but fail to exhibit airway hyperresponsiveness in both in vitro and in vivo assessments of lung function. Reconstitution of these deficient mice with the IL-10 gene fully restores development of airway hyperresponsiveness comparable to control mice. These results identify an important role of IL-10, downstream of the inflammatory cascade, in regulating the tone of the airways after allergic sensitization and challenge.


The specificity of alcohol dehydrogenase with cis-retinoids. Activity with 11-cis-retinol and localization in retina.
Studies in knockout mice support the involvement of alcohol dehydrogenases ADH1 and ADH4 in retinoid metabolism, although kinetics with retinoids are not known for the mouse enzymes. Moreover, a role of alcohol dehydrogenase (ADH) in the eye retinoid interconversions cannot be ascertained due to the lack of information on the kinetics with 11-cis-retinoids. We report here the kinetics of human ADH1B1, ADH1B2, ADH4, and mouse ADH1 and ADH4 with all-trans-, 7-cis-, 9-cis-, 11-cis- and 13-cis-isomers of retinol and retinal. These retinoids are substrates for all enzymes tested, except the 13-cis isomers which are not used by ADH1. In general, human and mouse ADH4 exhibit similar activity, higher than that of ADH1, while mouse ADH1 is more efficient than the homologous human enzymes. All tested ADHs use 11-cis-retinoids efficiently. ADH4 shows much higher k(cat)/K(m) values for 11-cis-retinol oxidation than for 11-cis-retinal reduction, a unique property among mammalian ADHs for any alcohol/aldehyde substrate pair. Docking simulations and the kinetic properties of the human ADH4 M141L mutant demonstrated that residue 141, in the middle region of the active site, is essential for such ADH4 specificity. The distinct kinetics of ADH4 with 11-cis-retinol, its wide specificity with retinol isomers and its immunolocalization in several retinal cell layers, including pigment epithelium, support a role of this enzyme in the various retinol oxidations that occur in the retina. Cytosolic ADH4 activity may complement the isomer-specific microsomal enzymes involved in photopigment regeneration and retinoic acid synthesis.


[Diagnostic potential of ultrasound examination for tumor recurrence and metastasis].
Ultrasound examination was carried out in 252 patients following specific treatment of uterine and ovarian malignancies. To compare the effectiveness of diagnostic procedures, transabdominal and transvaginal examinations were performed in succession in all the patients using ultrasound (Doppler) technique. Transvaginal study proved relatively more sensitive (98.9%) and diagnostically accurate (97.2%) in detecting relapse. Transabdominal examination carried out in conjunction with color Doppler imaging was best for metastasis detection.


Proteomic analyses uncover a new function and mode of action for mouse homolog of Diaphanous 2 (mDia2).
mDia2 is an auto-inhibited Formin influencing actin dynamics upon conversion to the active conformation. mDia2 regulates actin-based protrusions and cell invasion, cell differentiation, vesicle trafficking, and cytokinesis. However, whether mDia2 has additional functions and how its action is functionally specified remain unknown. Here we draw the interactome of auto-inhibited and constitutively active mDia2 to address these issues. We embed mDia2 in protein networks accounting for its attributed functions and unexpectedly link it to the Ubiquitin Proteasome System. Taking FBXO3 as a test case, we show that mDia2 binds FBXO3 and p53, and regulates p53 transcriptional activity in an actin-nucleation-independent and conformation-insensitive manner. Increased mDia2 and FBXO3 levels elevate p53 activity and expression thereby sensitizing cells to p53-dependent apoptosis, whereas their decrease produces opposite effects. Thus, we discover a new role of mDia2 in p53 regulation suggesting that the closed conformation is biologically active and an FBXO3-based mechanism to functionally specify mDia2's activity.


Racial differences in the incidence of chronic kidney disease.
The incidence of ESRD is higher in African Americans than in whites, despite reports of a similar or lower prevalence of CKD. This study compared the incidence of CKD among young African-American and white adults over 20 years of follow-up in the community-based Coronary Artery Risk Development in Young Adults study. Participants included 4119 adults, 18-30 years of age, with an estimated GFR (eGFR) ≥60 ml/min per 1.73 m(2) at baseline. Incident CKD was defined as an eGFR <60 ml/min per 1.73 m(2) and a ≥25% decline in eGFR at study visits conducted 10, 15, and 20 years after baseline. At baseline, the mean age of African Americans and whites was 24 and 26 years, respectively (P<0.001), and 56% and 53% of participants, respectively, were women (P=0.06). There were 43 incident cases of CKD during follow-up, 29 (1.4%) among African Americans and 14 (0.7%) among whites (P=0.02). The age- and sex-adjusted hazard ratio (HR) for incident CKD comparing African Americans to whites was 2.56 (95% confidence interval [95% CI], 1.35-5.05). After further adjustment for body mass index, systolic BP, fasting plasma glucose, and HDL cholesterol, the HR was 2.51 (95% CI, 1.25-5.05). After multivariable adjustment including albuminuria at year 10, the HR for CKD at year 15 or 20 was 1.12 (95% CI, 0.52-2.41). In this study, the 20-year CKD incidence was higher among African Americans than whites, a difference that is explained in part by albuminuria.


Protective effects of polydatin on experimental testicular torsion and detorsion injury in rats.
Oxidative stress plays a critical role in the process of testicular torsion and detorsion (T/D). The purpose of the present study was to investigate the protective effect of polydatin (PD) on testicular T/D injury. Rats were randomly divided into three groups, a sham group, a group subjected to 2h torsion followed by 24h detorsion and a group subjected to T/D and injected i.p. with 20mgkg-1 PD 30min before detorsion. Unilateral orchiectomy was performed after 24h of reperfusion. Half the testes were prepared for histological examination by haematoxylin-eosin staining and the terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling (TUNEL) technique. In the remaining tissues, levels of malondialdehyde (MDA), catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) were determined, as was the expression of several apoptosis-related proteins. Compared with the T/D group, PD pretreatment significantly ameliorated the morphological damage, lowered the Cosentino histological score and increased the mean number of germ cell layers and Johnsen's testicular biopsy score. In addition, PD treatment markedly decreased MDA levels and upregulated CAT, GPx and SOD activity. Furthermore, PD decreased T/D-induced germ cell-specific apoptosis, attenuated the activation of caspase-3, caspase-8, caspase-9 and poly(ADP-ribose) polymerase and increased the Bcl-2/Bax ratio. The findings indicate that PD has a protective effect against testicular T/D injuries, especially at the histological, antioxidative stress and antiapoptotic levels.


Organic cofactors in the metabolism of Dehalococcoides mccartyi strains.
Dehalococcoides mccartyi strains are strictly anaerobic organisms specialized to grow with halogenated compounds as electron acceptor via a respiratory process. Their genomes are among the smallest known for free-living organisms, and the embedded gene set reflects their strong specialization. Here, we briefly review main characteristics of published Dehalococcoides genomes and show how genome information together with cultivation and biochemical experiments have contributed to our understanding of Dehalococcoides physiology and biochemistry. We extend this approach by the detailed analysis of cofactor metabolism in Dehalococcoides strain CBDB1. Dehalococcoides genomes were screened for encoded proteins annotated to contain or interact with organic cofactors, and the expression of these proteins was analysed by shotgun proteomics to shed light on cofactor requirements. In parallel, cultivation experiments testing for vitamin requirements showed that cyanocobalamin (vitamin B12), thiamine and biotin were essential supplements and that cyanocobalamin could be substituted by dicyanocobinamide and dimethylbenzimidazole. Dehalococcoides genome analysis, detection of single enzymes by shotgun proteomics and inhibition studies confirmed the expression of the biosynthetic pathways for pyridoxal-5-phosphate, flavin nucleotides, folate, S-adenosylmethionine, pantothenate and nicotinic acids in strain CBDB1. Haem/cytochromes, quinones and lipoic acids were not necessary for cultivation or dechlorination activity and no biosynthetic pathways were identified in the genomes.


A molecular-capsular-type prediction system for 90 Streptococcus pneumoniae serotypes using partial cpsA-cpsB sequencing and wzy- or wzx-specific PCR.
In a previous study, a molecular capsular type (MCT) prediction system for 51 Streptococcus pneumoniae serotypes was developed based on a combination of partial cpsA-cpsB sequencing and serotype(s)/group(s)-specific PCR. In this study, another 169 S. pneumoniae isolates were added to the existing database of 427 isolates, including representatives of all 39 serotypes not previously studied. In addition to the authors' own limited sequence data for all 90 serotypes, cpsA-cpsB sequence data published by the S. pneumoniae capsular loci-sequencing group (http://www.sanger.ac.uk/Projects/S_pneumoniae/CPS/) at the Sanger Institute or available from GenBank were incorporated into the database. All serotypes, except 25A, were represented by at least two isolates. The number of sequence types identified was 138, of which 110 corresponded to single conventional serotypes (CSs); of these, 57 were represented by two or more isolates. Twenty-six sequence types were shared by between two and four CSs. To resolve these shared cpsA-cpsB sequence types and increase the discriminatory power of our system, the genes encoding the capsular polysaccharide flippase (wzx) and polymerase (wzy) were annotated and 24 new serotype(s)/group(s)-specific PCRs targeting wzy and two targeting wzx were designed. Using both cpsA-cpsB sequencing and wzx/wzy PCR, MCT correctly predicted the CSs of 516 (73 %) and the serogroup of an additional 155 (22 %) of the 708 isolates evaluated. For 5 % of isolates, MCT could not distinguish between members of five serotype pairs (37 isolates) containing members of different serogroups. Although further study of the relationship between MCT and CS is needed, this system now allows serotype or serogroup identification of 95 % of S. pneumoniae isolates.


Urocystolith detection: comparison of survey, contrast radiographic and ultrasonographic techniques in an in vitro bladder phantom.
Urocystoliths of 9 mineral types from 437 canine patients submitted to the University of Minnesota Urolith Bank were imaged in a urinary bladder phantom. Imaging techniques simulated were survey radiography, pneumocystography, double contrast cystography (two iodine concentrations) and real-time ultrasonography (3.5 MHz, 5.0 MHz, 7.5 MHz). Imaging techniques were compared for accuracy of urocystolith detection, accuracy of urocystolith enumeration, and tendencies for over or undercounting. Across urocystolith mineral types, the false negative rates (no urocystoliths detected in a given case) for survey radiographs range from 2 to 27%. Pneumocystographic techniques are one-half as likely to yield false negative results as are survey radiographic techniques. Underestimates of urocystolith numbers and false negatives are likely using 80 mg iodine/ml double contrast cystography because calcium-based urocystoliths are isopaque in this contrast medium dilution. The 200 mg iodine/ml double contrast cystographic techniques are unlikely to yield false negative diagnoses even for very small (< or = 1.0 mm) urocystoliths and is comparable to pneumocystography for detection and slightly better for enumeration. The likelihood of an ultrasonographic false negative for urocystoliths decreases with increasing MHz. Under optimal conditions using a 7.5 MHz mechanical sector transducer, the false negative rates were comparable to double contrast cystography, but rates increased notably with lower MHz transducers.


Integrating mechanistic and correlative niche models to unravel range-limiting processes in a temperate amphibian.
Insights into the causal mechanisms that limit species distributions are likely to improve our ability to anticipate species range shifts in response to climate change. For species with complex life histories, a mechanistic understanding of how climate affects different lifecycle stages may be crucial for making accurate forecasts. Here, we use mechanistic niche modeling (NicheMapR) to derive "proximate" (mechanistic) variables for tadpole, juvenile, and adult Rana temporaria. We modeled the hydroperiod, and maximum and minimum temperatures of shallow (30 cm) ponds, as well as activity windows for juveniles and adults. We then used those ("proximate") variables in correlative ecological niche models (Maxent) to assess their role in limiting the species' current distribution, and to investigate the potential effects of climate change on R. temporaria across Europe. We further compared the results with a model based on commonly used macroclimatic ("distal") layers (i.e., bioclimatic layers from WorldClim). The maximum temperature of the warmest month (a macroclimatic variable) and maximum pond temperatures (a mechanistic variable) were the most important range-limiting factors, and maximum temperature thresholds were consistent with the observed upper thermal limit of R. temporaria tadpoles. We found that range shift forecasts in central Europe are far more pessimistic when using distal macroclimatic variables, compared to projections based on proximate mechanistic variables. However, both approaches predicted extensive decreases in climatic suitability in southern Europe, which harbors a significant fraction of the species' genetic diversity. We show how mechanistic modeling provides ways to depict gridded layers that directly reflect the microenvironments experienced by organisms at continental scales, and to reconstruct those predictors without extrapolation under novel future conditions. Furthermore, incorporating those predictors in correlative ecological niche models can help shed light on range-limiting processes, and can have substantial impacts on predictions of climate-induced range shifts.


Bacterial cholecystitis with cardiac and pulmonary dissemination in a blue-naped mousebird (Urocolius macrourus).
A blue-naped mousebird (Urocolius macrourus) was diagnosed by gross necropsy and histopathology as having a chronic, fibrosing bacterial cholecystitis. Acute, severe, necrotizing pneumonia and myocarditis also were present with intralesional gram-negative bacteria. The bacteria within the lungs and heart were suspected to have spread from the biliary tract because of the pattern of distribution and similar gram-staining characteristics. Enterobacter sp. and Escherichia coli were cultured from the pulmonary lesions. Cloacal cultures in clinically normal blue-naped mousebirds and speckled mousebirds (Colius striatus) yielded both Enterobacter sp. and E. coli. We hypothesize a pathogenesis in this bird consisting of biliary stasis of unknown etiology and eventual infection of the biliary tract by the normal gram-negative gastrointestinal flora. Death was believed to be a result of cardiac and respiratory dysfunction secondary to the bacterial dissemination from the biliary tree and endotoxemia.


Multiomic profiling reveals metabolic alterations mediating aberrant platelet activity and inflammation in myeloproliferative neoplasms.
Platelets from patients with myeloproliferative neoplasms (MPNs) exhibit a hyperreactive phenotype. Here, we found elevated P-selectin exposure and platelet-leukocyte aggregates indicating activation of platelets from essential thrombocythemia (ET) patients. Single cell RNA-seq analysis of primary samples revealed significant enrichment of transcripts related to platelet activation, mTOR and oxidative phosphorylation (OXPHOS) in ET patient platelets. These observations were validated via proteomic profiling. Platelet metabolomics revealed distinct metabolic phenotypes consisting of elevated ATP generation, accompanied by increases in the levels of multiple intermediates of the tricarboxylic acid (TCA) cycle, but lower alpha-ketoglutarate (α-KG) in MPN patients. Inhibition of PI3K/AKT/mTOR signaling significantly reduced metabolic responses and hyperreactivity in MPN patient platelets, while α-KG supplementation markedly reduced oxygen consumption and ATP generation. Ex vivo incubation of platelets from both MPN patients and Jak2 V617F mice with α-KG significantly reduced platelet activation responses. Oral α-KG supplementation of Jak2 V617F mice decreased splenomegaly and reduced hematocrit, monocyte and platelet counts. Finally, α-KG incubation significantly decreased proinflammatory cytokine secretion from MPN CD14+ monocytes. Our results reveal a previously unrecognized metabolic disorder in conjunction with aberrant PI3K/AKT/mTOR signaling, contributing to platelet hyperreactivity in MPN patients.


Moderator effects of working memory on the stability of ADHD symptoms by dopamine receptor gene polymorphisms during development.
We tested the hypothesis that dopamine D1 and D2 receptor gene (DRD1 and DRD2, respectively) polymorphisms and the development of working memory skills can interact to influence symptom change over 10 years in children with attention-deficit/hyperactivity disorder (ADHD). Specifically, we examined whether improvements in working memory maintenance and manipulation from childhood to early adulthood predicted the reduction of ADHD symptoms as a function of allelic variation in DRD1 and DRD2. Participants were 76 7-11-year-old children with ADHD who were genotyped and prospectively followed for almost 10 years. ADHD symptoms were rated using the Attention Problems scale on the Child Behavior Checklist, and verbal working memory maintenance and manipulation, measured by Digit Span forward and backward, respectively, were assessed at baseline and follow-up. After correction for multiple testing, improvements in working memory manipulation, not maintenance, predicted reduction of symptomatology over development and was moderated by major allele homozygosity in two DRD1 polymorphisms (rs4532 and rs265978) previously linked with variation in D1 receptor expression. Depending on genetic background, developmental factors including age-dependent variation in DRD1 penetrance may facilitate the link between improvements in higher-order working memory and the remission of symptoms in individuals with childhood-diagnosed ADHD. Furthermore, the current findings suggest that DRD1 might contribute minimally to the emergence of symptoms and cognitive difficulties associated with ADHD in childhood, but may act as a modifier gene of these clinical features and outcome during later development for those with ADHD.


Eating disorders and gastrointestinal peptides.
Disturbances in gastrointestinal hormones have been implicated in the pathogenesis of eating disorders such as anorexia nervosa and bulimia nervosa. However, the contribution of these hormonal changes to the onset and maintenance of eating disorder remains unclear. We focus our review on a selective number of gastrointestinal hormones that are known to play a role in the regulation of short-term or long-term energy balance and examine their association with eating disorder in recently published literature. Several new studies reported differential changes of ghrelin isoforms during fasting and following nutrient ingestion. New findings on other appetite-regulating hormones (peptide YY, cholecystokinin, incretin hormones and pancreatic polypeptide) at different nutritional states and disease stage have also been reported in subtypes of eating disorder. Most of the changes in peripheral hormones disappeared or partially recovered after the restoration of weight with nutritional and behavioral therapy. Dysregulation of gastrointestinal hormones is more likely to contribute to the maintenance of the disordered eating behavior and related metabolic outcomes as well as the clinical course rather than causing them. A better understanding of this relationship also carries implications for developing targeted hormone-base treatment for eating disorder.


[ENT antibiotic therapy: therapeutic guidelines. Part I].
We discuss the variety of ear, nose and throat infections and then define the criteria for the choice of an adequate antibiotic. The importance of "supercalculated" therapy based on the results of Gram's stain is stressed, as it can save patients from over-treatment and adverse side-effects. Indications for Gram's stain in ENT as well as the value of the characteristics of the pus produced by different pathogens are described. The severity of the infection determines the choice between parenteral or oral therapy. The importance of the decision taken on the 1st and 3rd day of a severe infection is emphasised. Therapeutic principles in ENT infections are shown by grouping different infections with a similar spectrum of pathogens; also the indications for antibiotic therapy are outlined. Finally the priority of antibiotic treatment in some surgical infections is mentioned, and the peri-operative prophylaxis and treatment are described as well as the guidelines for treatment of infectious post-operative complications.


Nutritional status of mentally retarded children in northwest spain: II. Biochemical indicators.
To evaluate the nutritional status of mentally retarded children in the region of Galicia, northwest Spain, on the basis of serum biochemistry variables. The following serum biochemistry variables were determined in a sample of 128 mentally retarded children (81 boys and 47 girls): albumin, prealbumin, retinol-binding protein, transferrin, ferritin, ceruloplasmin, iron, calcium, phosphorus, copper, total cholesterol, HDL cholesterol, LDL cholesterol, triglycerides, alkaline phosphatase, transaminases and carnitine. A preliminary statistical analysis indicated that most information content could be maintained taking into consideration only five of these variables. A factor analysis of the resulting 5 x 128 variables-by-subjects data matrix was then performed, identifying three factors (FB1, FB2 and FB3) that together explained 74% of total variance. Taking these factors as indicators of nutritional status, the data were then analysed for possible effects of age, gender, socioeconomic and family environment, IQ, presence/absence of cerebral palsy, quality of diet, appetite and use of antiepileptics. The analysis suggests that most subjects were in the normal nutritional range, but that about 37% showed either borderline or definite malnutrition. The FB1 score showed a significant, positive correlation with age, while FB2 score was negatively correlated with age. The FB1 score showed a significant positive correlation with socioeconomic class, and was significantly higher among children from inland-rural areas than among children from coastal or urban areas. The FB1 score was also positively correlated with parents' mean age. Finally, the results suggest that antiepileptic treatment with phenobarbital or diphenylhydantoin led to reductions in serum levels of calcium and phosphorus. Malnutrition as revealed by biochemical variables is highly prevalent among mentally retarded children, and the prevalence is affected by socioeconomic and family environment variables.


No identification of abrupt onsets that capture attention: evidence against a unified model of spatial attention.
Many studies have reported that spatial attention can be involuntarily captured by salient stimuli such as abrupt onsets. These involuntary shifts are often assumed to have the same effects on feature extraction as voluntary shifts: there are two different ways of moving the same attentional mechanism. According to this unified model of spatial attention, all shifts of attention should enhance the identification of attended objects. We directly tested this assumption using compatibility effects in a series of spatial cueing experiments. Participants searched a display and indicated whether the target number was greater or less than five. The salient precues were also numbers, allowing measurement of compatibility effects between the precue and the target. Precues that reliably predicted the target location produced compatibility effects (e.g., the precue "1" facilitated responding to the target "one"), indicating enhanced identification of the precue. Compatibility effects were also found for precues that were nonpredictive but had the target-finding feature (i.e., contingent capture). Critically, however, four separate experiments failed to find compatibility effects for salient abrupt onsets that were neither predictive nor task-relevant. This is surprising given that these same precues produced enormous cue validity effects (up to 186 ms), suggesting salience-based attention capture. Our findings argue against the unified model: salience-based attention capture recruits different attentional mechanisms than contingent capture or voluntary shifts in attention.


Analysis of the in vivo phosphorylation state of protein phosphatase inhibitor-2 from rabbit skeletal muscle by fast-atom bombardment mass spectrometry.
A new procedure has been developed for identifying phosphoserine residues in proteins, and is used to analyse the in vivo phosphorylation state of inhibitor-2. The method employs reverse-phase liquid chromatography to resolve phosphorylated and dephosphorylated forms of peptides and fast-atom bombardment mass spectrometry (FABMS) to identify phosphorylated derivatives. The positions of phosphorylation sites within peptides are located by gas-phase sequencer analysis after conversion of phosphoserine residues to S-ethylcysteine. The phosphorylation sites on inhibitor-2 were identified as serines-86, -120 and -121, the three residues phosphorylated in vitro by casein kinase-II. Serine-86 was phosphorylated to 0.7 mol/mol and serines-120 and -121 each to 0.3 mol/mol. These values were not altered significantly by intravenous injection of adrenalin or insulin. No phosphate was present in the region comprising residues 1-49, even after injection of adrenalin, demonstrating that inhibitor-2 is not a substrate for cyclic AMP-dependent protein kinase in vivo. The absence of phosphotyrosine also indicated that inhibitor-2 is not a physiological substrate for the insulin receptor. Surprisingly, no phosphate was present at threonine-72, the residue phosphorylated in vitro by glycogen synthase kinase-3, after injection of either propranolol, adrenalin or insulin. The implications of this finding for the in vivo activation of protein phosphatase 1I (the 1:1 complex between inhibitor-2 and the catalytic subunit of protein phosphatase-1) are discussed. FABMS analysis of inhibitor-2 confirmed the accuracy of the primary structure reported previously, and showed that the only post-translational modifications were an N-acetyl moiety and the three phosphoserine residues. FABMS also demonstrated the presence of an additional serine residue at the C-terminus, and showed that 50% of isolated inhibitor-2 molecules lack the C-terminal Ser-Ser dipeptide.


Dental pulp cells cocultured with macrophages aggravate the inflammatory conditions stimulated by LPS.
Pulp inflammation is complex interactions between different types of cells and cytokines. To mimic the interactions of different types of cells in inflamed dental pulp tissues, dental pulp cells (DPCs) were cocultured with different ratios of macrophages (THP-1) or LPS treatment. DPCs were cocultured with various ratios of THP-1, then photographed cell morphology and determined cell viability by MTT assay at preset times. Total RNA was also extracted to measure the inflammation marker-IL-6 and IL-8 expressions by RT-Q-PCR. The DPCs and THP-1 were treated with 0.01 - 1μg/ml lipopolysaccharide (LPS) and extract RNA at preset times, and detected IL-6 and IL-8 expression. DPCs were cocultured with various ratios of THP-1 with 0.1 μg/mL LPS, and detected IL-6 and IL-8 expression after 24 and 48 h. The data were analyzed by unpaired t-test or Mann-Whitney test. Differences were considered statistically significant when p < 0.05. THP-1 and DPCs coculture models did not suppress the viability of DPCs and THP-1. Cocultured with various ratios of THP-1 could increase IL-6 and IL-8 expressions of DPCs (p = 0.0056 - p < 0.0001). The expressions of IL-6 and IL-8 were stronger in higher ratio groups (p = 0.0062 - p < 0.0001). LPS treatment also induced IL-6 and IL-8 expressions of DPCs and THP-1 (p = 0.0179 - p < 0.0001 and p = 0.0189 - p < 0.0001, separately). Under the presence of 0.1 μg/mL LPS, DPCs cocultured with THP-1 for 24 h also enhanced IL-6 and IL-8 expression (p = 0.0022). After cocultured with a higher ratio of THP-1 for 48 h, IL-6 and IL-8 expressions were even stronger in the presence of LPS (p = 0.0260). Coculturing dental pulp cells and macrophages under LPS treatment aggravate the inflammatory process. The responses of our models were more severe than traditional inflamed dental models and better represented what happened in the real dental pulp. Utilizing our models to explore the repair and regeneration in endodontics will be future goals.


[Properties of muscle glycerol-3-phosphate dehydrogenase in rabbits with Brown-Pearce carcinoma].
Specific activity of glycerol-3-phosphate dehydrogenase, estimated in direct and reverse reactions, was decreased, as compared with normal state, without any alterations in the ratio of activities, in muscle homogenates of rabbits with Brown-Pearce carcinoma. The enzymatic activity was decreased by an order of magnitude in homogenates of the tumor as compared with muscle homogenates of the tumor-bearing animals simultaneously with relative stimulation of the reverse reaction. Specific activity of the crystalline enzyme from muscles of tumor-bearing animals was decreased as compared with normal tissues but the ratio of activities was 2-fold increased due to the reverse reaction. In the carcinoma activity of one of the three enzyme fractions, obtained after isoelectrofocusing, was similar to the control values; in the second fraction the ratio of activities was decreased due to inhibition of the direct reaction, the third fraction was inactive. Amino acid composition of crystalline glycerol-3-phosphate dehydrogenase, isolated from tumor-bearing animals, was different in content of 5 amino acids from the enzyme amino acid composition of controls, thus suggesting the structural differences of the proteins from these fractions.


Adenocarcinoma of the jejunum in an 18-year-old patient: a case report.
The clinical history and surgical findings of an adenocarcinoma of the jejunum in an 18-year-old black man are recorded. The world literature on this subject has been reviewed, revealing a total of six cases of adenocarcinoma of the jejunum and ileum in patients under 20 years of age. This seventh case is recorded for its rarity and the findings which should alert the clinician: cramping abdominal pain, weight loss, and occult blood in the stool.


ATP-stimulated endoprotease is associated with the cell membrane of E. coli.
Despite knowledge of the physiological significance and regulation of protein degradation in bacteria, the pathway of proteolysis and the responsible enzymes are still not known. Degradation of cell proteins in bacterial and animal cells requires continuous ATP production, inhibition of which in Escherichia coli prevents the degradation of normal proteins in growing cells, accelerated breakdown of such proteins in starving cultures and the very rapid breakdown of abnormal proteins. Intracellular proteolysis proceeds by repeated endoproteolytic steps and ATP is required for the initial cleavages of the substrate. We have recently demonstrated ATP stimulation of proteolysis in extracts of bacterial and animal cells. These ATP-stimulated systems seem to be responsible for the rapid degradation of abnormal proteins in vivo, but they may also be involved in the catabolism of normal cell proteins, limited proteolysis, such as the processing of precursors for secreted or membrane proteins, and the selective inactivation of specific proteins, as occurs in the ATP-dependent cleavage of the lambda repressor by the recA protein. We report here that membrane fragments contain an ATP-stimulated protease that degrades cell proteins to large peptides (of molecular weight (MW) 71,500) which are then rapidly hydrolysed to amino acids by soluble ATP-independent enzymes.


Flavour stability of sterilised chickpeas stored in pouches.
The increasing need for sustainable food choices places a demand on developing palatable foods from lower impact production and with a suitable shelf-life. In this context, knowledge of the sensory properties of whole sterilised chickpeas is required to be able to make them more attractive to the consumers. The sensory quality of chickpeas is largely dependent on the aroma and flavour, which can be influenced by storage conditions. In this study, sterilised chickpeas in two different packaging materials with different oxygen permeabilities, stored up to 52 weeks at ambient temperature (20 °C) were investigated using both descriptive sensory profiling and gas chromatography - mass spectrometry - olfactometry analysis (GC-MS-O). During storage, intense, sulphury and meat-like aromas decreased in intensity, while with longer storage time hay-like, green-like and potato-like flavours and aromas became more apparent. A total of 40 odour-active volatiles were detected, of which some had properties reminiscent of the chickpea flavour profiles. However, no clear relationships could be established between these odorants and the sensory changes observed during storage by descriptive sensory analysis. It was concluded that, significant changes in the sensory attributes of sterilised chickpeas occurred during 52 weeks of storage, but that packaging material does not seem to largely influence the sensory attributes during storage.


Disintegrins from hematophagous sources.
Bloodsucking arthropods are a rich source of salivary molecules (sialogenins) which inhibit platelet aggregation, neutrophil function and angiogenesis. Here we review the literature on salivary disintegrins and their targets. Disintegrins were first discovered in snake venoms, and were instrumental in our understanding of integrin function and also for the development of anti-thrombotic drugs. In hematophagous animals, most disintegrins described so far have been discovered in the salivary gland of ticks and leeches. A limited number have also been found in hookworms and horseflies, and none identified in mosquitoes or sand flies. The vast majority of salivary disintegrins reported display a RGD motif and were described as platelet aggregation inhibitors, and few others as negative modulator of neutrophil or endothelial cell functions. This notably low number of reported disintegrins is certainly an underestimation of the actual complexity of this family of proteins in hematophagous secretions. Therefore an algorithm was created in order to identify the tripeptide motifs RGD, KGD, VGD, MLD, KTS, RTS, WGD, or RED (flanked by cysteines) in sialogenins deposited in GenBank database. The search included sequences from various blood-sucking animals such as ticks (e.g., Ixodes sp., Argas sp., Rhipicephalus sp., Amblyommasp.), tabanids (e.g., Tabanus sp.), bugs (e.g., Triatoma sp., Rhodnius prolixus), mosquitoes (e.g., Anopheles sp., Aedes sp., Culex sp.), sand flies (e.g., Lutzomyia sp., Phlebotomus sp.), leeches (e.g., Macrobdella sp., Placobdella sp.) and worms (e.g., Ancylostoma sp.). This approach allowed the identification of a remarkably high number of novel putative sialogenins with tripeptide motifs typical of disintegrins (>450 sequences) whose biological activity remains to be verified. This database is accessible online as a hyperlinked worksheet and displays biochemical, taxonomic, and gene ontology aspects for each putative disintegrin. It is also freely available for download (right click with the mouse) at links http://exon.niaid.nih.gov/transcriptome/RGD/RGD-Peps-WEB.xlsx (web version) and http://exon.niaid.nih.gov/transcriptome/RGD/RGD-sialogenins.zip (stand alone version).


Using bioinformatic protein sequence similarity to investigate if SARS CoV-2 infection could cause an ocular autoimmune inflammatory reactions?
Although severe acute respiratory syndrome coronavirus 2 (SARS CoV-2) infection have emerged globally, findings related to ocular involvement and reported cases are quite limited. Immune reactions against viral infections are closely related to viral and host proteins sequence similarity. Molecular Mimicry has been described for many different viruses; sequence similarities of viral and human tissue proteins may trigger autoimmune reactions after viral infections due to similarities between viral and human structures. With this study, we aimed to investigate the protein sequence similarity of SARS CoV-2 with retinal proteins and retinal pigment epithelium (RPE) surface proteins. Retinal proteins involved in autoimmune retinopathy and retinal pigment epithelium surface transport proteins were analyzed in order to infer their structural similarity to surface glycoprotein (S), nucleocapsid phosphoprotein (N), membrane glycoprotein (M), envelope protein (E), ORF1ab polyprotein (orf1ab) proteins of SARS CoV-2. Protein similarity comparisons, 3D protein structure prediction, T cell epitopes-MHC binding prediction, B cell epitopes-MHC binding prediction and the evaluation of the antigenicity of peptides assessments were performed. The protein sequence analysis was made using the Pairwise Sequence Alignment and the LALIGN program. 3D protein structure estimates were made using Swiss Model with default settings and analyzed with TM-align web server. T-cell epitope identification was performed using the Immune Epitope Database and Analysis (IEDB) resource Tepitool. B cell epitopes based on sequence characteristics of the antigen was performed using amino acid scales and HMMs with the BepiPred 2.0 web server. The predicted peptides/epitopes in terms of antigenicity were examined using the default settings with the VaxiJen v2.0 server. Analyses showed that, there is a meaningful similarities between 6 retinal pigment epithelium surface transport proteins (MRP-4, MRP-5, RFC1, SNAT7, TAUT and MATE) and the SARS CoV-2 E protein. Immunoreactive epitopic sites of these proteins which are similar to protein E epitope can create an immune stimulation on T cytotoxic and T helper cells and 6 of these 9 epitopic sites are also vaxiJen. These result imply that autoimmune cross-reaction is likely between the studied RPE proteins and SARS CoV-2 E protein. The structure of SARS CoV-2, its proteins and immunologic reactions against these proteins remain largely unknown. Understanding the structure of SARS CoV-2 proteins and demonstration of similarity with human proteins are crucial to predict an autoimmune response associated with immunity against host proteins and its clinical manifestations as well as possible adverse effects of vaccination.


Development and Characterization of Olanzapine Loaded Poly(lactide-coglycolide) Microspheres for Depot Injection: In vitro and In vivo Release Profiles.
The purpose of this study was to develop a new PLGA based microsphere formulation aimed to release the olanzapine for the period of one month which will result in increased compliance. Microspheres loaded with olanzapine were prepared using oil in water emulsion and solvent evaporation technique. The microspheres were characterized by surface morphology, shape, size, bulk density, encapsulation efficiency, and Fourier transform infrared spectrometry. In vitro release studies were performed in phosphate buffer at 37°C and in vivo studies were conducted on male Sprague- Dawley rats. The morphological results indicated that microspheres produced were having a smooth surface, spherical shape and the size in the range from 9.71 to 19.90 μm mean diameter. Encapsulation efficiency of olanzapine loaded microspheres was in the range of 78.53 to 96.12% and was affected by changing the ratio of lactic to glycolic acid in copolymer PLGA. The properties of PLGA and other formulation parameters had a significant impact on in vitro and in vivo release of drug from microspheres. In vitro release kinetics revealed that release of drug from microspheres is by both non-Fickian diffusion and erosion of PLGA polymer. In vivo data indicated an initial burst release and then sustained release depending on properties of PLGA, microsphere size, and bulk density. This study indicates that microsphere formulations developed with PLGA (75:25) and PLGA (85:15) have provided a sufficient steady release of drug for at least 30 days and can be potential candidates for 30-day depot injection drug delivery of olanzapine.


Effect of 2-n-heptyl-4-hydroxyquinoline N-oxide on proton permeability of the mitochondrial membrane.
The respiratory-chain inhibitor 2-n-heptyl-4-hydroxyquinoline N-oxide catalyses transmembrane proton transport driven by a pH gradient in isolated rat liver mitochondria. This effect explains the apparent blockade of net proton translocation by this compound in mitochondria respiring with ferrocyanide as described by Papa, Lorusso, Guerrieri, Boffoli, Izzo & Capuano [(1977) in Bioenergetics of Membranes (Packer, Papageorgiu & Trebst, eds.), pp. 377-388, Elsevier/North-Holland, Amsterdam] and by Lorusso, Capuano, Boffoli, Stefanelli & Papa [(1979) Biochem. J. 182, 133-147].


Effects of vitamin D supplementation on blood pressure.
Inconsistent findings from epidemiological studies have continued the controversy over the role of oral vitamin D supplementation in reducing blood pressure in normotensive or hypertensive populations. We performed a literature search up to December 2009, with no restrictions. Only double-blind randomized controlled trials (RCTs) of oral vitamin D supplementation in normotensive or hypertensive individuals with blood pressure measurements were included. From 244 retrieved papers, four RCTs involving 429 participants met our inclusion criteria for this meta-analysis. Vitamin D supplementation reduced systolic blood pressure (SBP) by 2.44 mm Hg (weighted mean difference [WMD]: -2.44, 95% confidence interval [CI]: -4.86, -0.02), but not diastolic blood pressure (DBP) (WMD: -0.02, 95% CI: -4.04, 4.01) compared with calcium or placebo. Subgroup analysis suggested that the change of blood pressure did not vary markedly across the dose of vitamin D supplementation, study length, or intervention. Oral vitamin D supplementation may lead to a reduction in systolic blood pressure but not diastolic blood pressure. Given the small number of trials and small but statistically significant reduction in systolic blood pressure from this meta-analysis, further studies are required to confirm the magnitude of the effect of vitamin D on blood pressure reduction and define the optimum dose, dosing interval, and type of vitamin D to administer.


Steroidogenic effect of atrial natriuretic factor in isolated mouse Leydig cells is mediated by cyclic GMP.
The effects of different atrial natriuretic peptides on cyclic GMP formation and steroidogenesis have been studied in Percoll-purified mouse Leydig cells. Rat atrial peptides rANP (rat atrial natriuretic peptide), rAP-I (rat atriopeptin I) and rAP-II (rat atriopeptin II), in the presence of a phosphodiesterase inhibitor, stimulated cyclic GMP formation in a concentration-dependent manner. In the presence of saturating concentrations of the peptides, a 400-600 fold stimulation of cyclic GMP accumulation was observed. Among the peptides, rAP-II appeared to be the most potent. ED50 values (concentration causing half-maximal effect) for rAP-II, rANP and rAP-I were 1 X 10(-9) M, 2 X 10(-9) M and 2 X 10(-8) M respectively. A parallel stimulation of cyclic GMP formation and testosterone production by the cells was observed after incubation of the cells with various concentrations of rAP-II. In the presence of a saturating concentration of rAP-II (2 X 10(-8) M), maximum stimulation of intracellular cyclic GMP content was obtained within 5 min of incubation. Testosterone production by mouse Leydig cells could be stimulated by 8-bromo cyclic GMP in a concentration-related manner. At a 10 mM concentration of the cyclic nucleotide, steroidogenesis was stimulated to a similar extent as that obtained with a saturating concentration of human chorionic gonadotrophin (5 ng/ml). On the basis of these results we conclude that cyclic GMP acts as a second messenger in atrial-peptide-stimulated steroidogenesis in mouse Leydig cells. The steroidogenic effect of atrial peptides appears to be species-specific, since none of these peptides stimulated testosterone production by purified Leydig cells of rats, though in these cells a 40-60-fold stimulation of cyclic GMP formation in response to each of the three peptides was observed. However, 8-bromo cyclic GMP could stimulate testosterone production in rat Leydig cells. Therefore we conclude that the lack of steroidogenic response in rat Leydig cells to atrial-natriuretic-factor-stimulation results from an insufficient formation of cyclic GMP in these cells. This species difference would appear to result from a lower guanylate cyclase activity in rat Leydig cells.


Multicenter trial to evaluate the safety and potential efficacy of pooled human fibrin sealant for the treatment of burn wounds.
The primary purpose of this multicenter study was to evaluate the safety and potential efficacy of a solvent/detergent-treated commercial fibrin sealant (human) for topical hemostasis in skin grafting. The study involved a prospective evaluation of changes in viral titers in patients with burns less than 15% after treatment with fibrin sealant (human). Each patient served as his/her own control for an unblinded, randomized comparison of donor site hemostasis and healing. Preoperative serum was obtained to screen for viral titers. At autografting, the recipient site and one of two randomly chosen donor sites were treated with fibrin sealant (human). The use of other hemostatic agents, including epinephrine was prohibited. Each donor site was covered with gauze to collect blood for estimation of the relative amount of bleeding. The healing of the graft and donor sites was observed. Viral titers and wounds were checked monthly for 6 months, and at 9 and 12 months postoperatively. Viral titers for human immunodeficiency virus; hepatitis A, B, and C; Epstein-Barr virus; and cytomegalovirus were obtained before and after treatment. Of 47 patients, 34 completed the full year of observation. After treatment, there were no seroconversions to any of the aforementioned viruses. Bleeding at the recipient site appeared well controlled with fibrin sealant (human). Although investigators felt that fibrin sealant (human) improved donor site hemostasis, differences in hemoglobin measurements of blood-soaked dressings failed to reach significance. No differences were noted with regard to acceleration of donor site healing, graft take, or scar maturation at the two groups of donor sites. Anecdotally, the maturation of the recipient site appeared to be accelerated. Fibrin sealant (human) is safe for use during excision and grafting, and its topical hemostatic potential needs to be examined in patients with larger burns. Its role in scar maturation also needs to be investigated.


Primary anaplastic large cell lymphoma of the central nervous system: prognostic effect of ALK-1 expression.
Anaplastic large cell lymphoma (ALCL) rarely occurs in the central nervous system. Although defined by its composition of large, pleomorphic, CD30-positive lymphocytes, ALCL is heterogeneous. Most are T cell but some are null cell. Most but not all have a characteristic 2:5 translocation producing the fusion protein ALK-1, which is reliably detected by immunohistochemistry. In systemic ALCL, ALK-1 expression correlates with young patient age and a favorable prognosis. Herein we report four new cases of primary central nervous system ALCL from the Mayo Clinic and incorporate additional data from five previously published cases. ALK-1 expression was determined in all nine tumors. Patient age was 4-66 years (mean 29 years) with a bimodal distribution: 6 < or = 22 years, 3 > or = 50 years. Six were female. Tumors were mostly supratentorial, five were multifocal, and seven had involvement of dura or leptomeninges. Seven tumors were T cell, two were null cell, and five of nine were ALK-1 immunopositive. Total mortality was six of nine. Three patients, 4-18 years of age (mean 13 years), were alive at 4.8-6.1 years postdiagnosis; these tumors were all ALK positive. Five patients, 13-66 years of age (mean 43 years), died of tumor 4 days to 11 weeks postdiagnosis; four of five of these tumors were ALK negative. One 10-year-old child with an ALK-positive tumor died of sepsis, but in remission. Central nervous system ALCL is aggressive. Our study suggests that a better outcome may be associated with young age and ALK-1 positivity, prognostic parameters similar to systemic ALCL.


Mesure de la réflectance de miroirs métalliques dans l'infrarouge.
The performance of high power CO(2) lasers depends directly on the quality of the metallic mirrors used. The mirrors are therefore the key elements of these lasers and must have a reflectance as near as possible to unity. Thus, we have to characterize these mirrors precisely, that is to say, to use suitable measurement methods to determine their optical properties with enough accuracy. In this paper we describe a reflectance measurement system based on the use of a classical spectrophotometer. This method gives relative measurements, but with the help of a known standard reflectance, we can obtain the absolute value. Experimental results show that the accuracy is ~1-2.5 x 10(-4) provided that many measurements are made and precautions are taken. We propose here to study these conditions and in particular to give the reflectances of metallic mirrors at 10.6 microm.


Stable and Efficient PtRu Electrocatalysts Supported on Zn-BTC MOF Derived Microporous Carbon for Formic Acid Fuel Cells Application.
Highly efficient, well-dispersed PtRu alloy nanoparticles supported on high surface area microporous carbon (MPC) electrocatalysts, are prepared and tested for formic acid oxidation reaction (FAOR). The MPC is obtained by controlled carbonization of a zinc-benzenetricarboxylate metal-organic framework (Zn-BTC MOF) precursor at 950°C, and PtRu (30 wt.%) nanoparticles (NPs) are prepared and deposited via a polyol chemical reduction method. The structural and morphological characterization of the synthesized electrocatalysts is carried out using powder X-ray diffraction (PXRD), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), transmission electron microscopy (TEM), an energy dispersive X-ray (EDX) technique, and gas adsorption analysis (BET). The FAOR performance of the catalysts is investigated through cyclic voltammetry (CV), chronoamperometry (CA), and electrochemical impedance spectroscopy (EIS). A correlation between high electrochemical surface area (ECSA) and high FAOR performance of the catalysts is observed. Among the materials employed, Pt1Ru2/MPC 950 with a high electrochemical surface area (25.3 m2 g-1) consequently showed superior activity of the FAOR (I r = 9.50 mA cm-2 and J m = 2,403 mA mg Pt - 1 ) at room temperature, with improved tolerance and stability toward carbonaceous species. The superior electrochemical performance, and tolerance to CO-poisoning and long-term stability is attributed to the high surface area carbon support (1,455 m2 g-1) and high percentage loading of ruthenium (20 wt.%). The addition of Ru promotes the efficiency of electrocatalyst by offering FAOR via a bifunctional mechanism.


Potentials of Aloe barbadensis inclusion in fish feeds on resilience to Aeromonas hydrophila infection in freshwater fish Labeo rohita.
Aquatic bacterial pathogens can cause severe economic loss in aquaculture industry. An opportunistic pathogen, Aeromonas hydrophila is responsible for Motile Aeromonas Septicemia, leading to high mortality rates in fish. The present study was focused on the efficacy of Aloe barbadensis replacing fishmeal diets on hematological, serum biochemical, antioxidant, histopathological parameters, and disease resistance against A. hydrophila infection in Labeo rohita. Isonitrogenous fishmeal replaced diets (FMR) were prepared with varying levels of A. barbadensis at D1 (0%) (control), D2 (25%), D3 (50%), D4 (75%) and D5 (100%) then fed to L. rohita. After 60 days of post-feeding, the experimental fish were challenged with A. hydrophila. Blood and organs were collected and examined at 1- and 15-days post infection (dpi). The results demonstrated that on 1 dpi, white blood cells (WBC), total protein, cholesterol and low-density lipoprotein (LDL) levels were significantly increased in D3 diet fed groups. The D2 and D3 diet fed group showed decreasing trends of serum glutamic pyruvic transaminase (SGPT) and antioxidant enzymes activity on 15 dpi. The histopathological architecture results clearly illustrated that the D3 diet fed group had given a higher protective effect by reducing the pathological changes associated with A. hydrophila infection in liver, intestine and muscle. Higher percentage of survival rate was also observed in D3 diet fed group. Therefore, the present study suggested that the dietary administration of A. barbadensis up to 50% fishmeal replacement (D3 diet) can elicit earlier antioxidant activity, innate immune response and improve survival rate in L. rohita against A. hydrophila infection.


[Extended spectrum beta-lactamase types in Klebsiella pneumoniae strains isolated from blood cultures].
The aim of this study was to investigate and type the extended spectrum beta-lactamases (ESBL) in Klebsiella pneumoniae strains isolated from blood cultures. Following the detection of antibiotic susceptibilities in 32 K. pneumoniae isolates, ESBL were detected in 13 (41%) of them by using double disc synergy test. Minimum inhibition concentrations for ceftazidime, cefotaxime, and aztreonam of ESBL positive strains were determined by E-test. After the extraction of the enzymes, the types of ESBLs were investigated by isoelectric focusing method. It was seen that, of all ESBL positive strains, one strain had four bands, one had three bands, six strains had two bands, and each of the others had only one beta-lactamase band. The results of polymerase chain reaction (PCR) revealed bla(SHV) in ten samples, bla(TEM) in six samples, and bla(SHV) with bla(TEM) in four samples. Ten SHV enzymes were typed as ESBL by PCR-RFLP (restriction fragment lenght polymorphism) method. The results of isoelectric focusing, PCR and RFLP performed in these ESBL positive K. pneumoniae isolates showed that the ESBL types could be SHV-2, SHV-5 and SHV-12 in the tested strains. It should always be taken into consideration that K. pneumoniae isolates could produce ESBLs and antibiotic treatment protocols should be adjusted in accordance.


[Nonlinear partial least squares method for simultaneous derivative spectrophotometric determination of two components in compound sulfamethoxazole].
A nonlinear model of consistency with derivative absorption in multiple components systems has been given. Model data were estimated by partial least squares method. A method for the simultaneous derivative spectrophotometric determination of two components with nonlinear partial least squares method was established. The method has been applied to the simultaneous determination of two components in compound sulfamethoxazole tablets. The average recoveries of sulfamethoxazole and trimethoprim in synthetic samples were 99.8% and 100.1%, and relative standard deviations were 1.3% and 1.6%, respectively. The results by nonlinear PLS method are significantly better than those by linear PLS method. The results for actual compound formulation agreed with those obtained by standard method.


Time course of smooth muscle cell proliferation in the intima and media of arteries following experimental angioplasty.
Smooth muscle cell (SMC) proliferation is known to be an important factor for the development of restenosis after percutaneous transluminal coronary angioplasty. To determine the time course of intimal and medial SMC proliferation and morphological changes after experimental angioplasty, an intimal atheroma was produced with repeated weak electrical stimulations in the right carotid artery of 45 male New Zealand White rabbits. Angioplasty was subsequently performed in 35 rabbits, and the proliferative responses were analyzed with histomorphological and immunohistological criteria at 3, 7, 14, 21, 28, and 42 days after intervention. A hemodynamic relevant stenosis after angioplasty was found in eight (23%) of 35 dilated arteries. In five rabbits the stenosis was due to a mural thrombus, and in three animals restenosis was caused by intimal SMC proliferation. In all dilated arteries the intimal wall thickness increased from 13 +/- 5 intimal cell layers (after electrical stimulation) to 33 +/- 14 cell layers during 28 days after angioplasty (p less than 0.05). Later than 4 weeks after angioplasty, no additional increase of intimal thickening occurred. Application of bromodeoxyuridine 18 and 12 hours before excision of the vessels allowed determination of the percent of cells undergoing DNA synthesis in the intima and media using monoclonal antibody against bromodeoxyuridine. SMCs were identified by alpha-actin staining. Immunohistological quantification of intimal SMC proliferation showed a maximum of cells undergoing DNA synthesis within the first 7 days after angioplasty (p less than 0.01). In contrast, medial proliferation of SMCs was delayed and showed a small but significant increase 21 days after dilatation (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Relationship Between Perineural Invasion in Prostate Needle Biopsy Specimens and Pathologic Staging After Radical Prostatectomy.
Prostate cancer is the second most common malignancy among men worldwide and the sixth cause of cancer-related death. Some authors have reported a relationship between perineural invasion (PNI), Gleason score, and the invasion of peripheral organs during prostatectomy. However, it is not yet clear whether pathological evidence of PNI is necessary for risk stratification in selecting treatment type. The clinical and pathological stages of prostate cancer are compared in patients under radical prostatectomy and in patients without perineural invasion. This cross-sectional study was conducted using a sample of 109 patients who attended a tertiary health care center from 2008 to 2013. The selection criteria were PNI in prostate biopsy with Gleason scores less than six, seven, and eight to ten. The participants were enrolled in a census manner, and they underwent clinical staging. After radical prostatectomy, the rates of pathological staging were compared. The under-staging and over-staging rates among those with and without perineural invasion in biopsy samples were compared. The concordance between Gleason scores according to biopsy and pathology was 36.7% (40 subjects). The concordance rate was 46.4% and 33.3% among those with and without PNI, respectively. The concordance rates were significantly varied in different subclasses of Gleason scores in patients without PNI (P = 0.003); the highest concordance rate was a Gleason score of 7 (63.6%) and the lowest was a Gleason score of eight to ten (25%). However, there were no significant differences in patients with PNI (P > 0.05). Although the presence of PNI in prostate biopsy is accompanied by higher surgical stages, PNI is not an appropriate independent factor in risk stratification.


Fundamental modeling of wave propagation in temporally relaxing media with applications to cardiac shear wave elastography.
Shear wave elastography (SWE) might allow non-invasive assessment of cardiac stiffness by relating shear wave propagation speed to material properties. However, after aortic valve closure, when natural shear waves occur in the septal wall, the stiffness of the muscle decreases significantly, and the effects of such temporal variation of medium properties on shear wave propagation have not been investigated yet. The goal of this work is to fundamentally investigate these effects. To this aim, qualitative results were first obtained experimentally using a mechanical setup, and were then combined with quantitative results from finite difference simulations. The results show that the amplitude and period of the waves increase during propagation, proportional to the relaxation of the medium, and that reflected waves can originate from the temporal stiffness variation. These general results, applied to literature data on cardiac stiffness throughout the heart cycle, predict as a major effect a period increase of 20% in waves propagating during a healthy diastolic phase, whereas only a 10% increase would result from the impaired relaxation of an infarcted heart. Therefore, cardiac relaxation can affect the propagation of waves used for SWE measurements and might even provide direct information on the correct relaxation of a heart.


[Three-dimensional morphological features of temporomandibular joint in skeletal malocclusion Class III patients with different vertical skeletal facial types].
To investigate three-dimensional morphological features of temporomandibular joint in skeletal malocclusion Class III with different vertical skeletal facial types. Methods: A total of 46 adults with skeletal malocclusion Class III were enrolled and allocated into a normal-angle, a high-angle and a low-angle groups. All patients were undergone cone beam computed tomography (CBCT). Images of temporomandibular joint were reconstructed and analyzed by Invivo 5. Nine measurements relevant to the positions and forms of condyle as well as joint spaces were performed. Results: Significant differences were not observed between the 2 sides of each group among skeletal malocclusion Class III (P>0.05). Compared with the high-angle and normal-angle patients, the low-angle patients had larger superior spaces, bigger gradients of the articular tublecle, longer condyle, and larger internal spaces between the condyle and the fossa. Compared with the low-angle and normal-angle patients, the high-angle patients had smaller condylar antero-posterior diameters. Conclusion: Class III malocclusion patients with different vertical skeletal patterns have different positions and forms of condyles, which should be paid attention to in clinic.