Conformational averaging in structural biology: issues, challenges and computational solutions.
Most experimental methods in structural biology provide time- and ensemble-averaged signals and, consequently, molecular structures based on such signals often exhibit only idealized, average features. Second, most experimental signals are only indirectly related to real, molecular geometries, and solving a structure typically involves a complicated procedure, which may not always result in a unique solution. To what extent do such conformationally-averaged, non-linear experimental signals and structural models derived from them accurately represent the underlying microscopic reality? Are there some structural motifs that are actually artificially more likely to be "seen" in an experiment simply due to the averaging artifact? Finally, what are the practical consequences of ignoring the averaging effects when it comes to functional and mechanistic implications that we try to glean from experimentally-based structural models? In this review, we critically address the work that has been aimed at studying such questions. We summarize the details of experimental methods typically used in structural biology (most notably nuclear magnetic resonance, X-ray crystallography and different types of spectroscopy), discuss their individual susceptibility to conformational (motional) averaging, and review several theoretical approaches, most importantly molecular dynamics simulations that are increasingly being used to aid experimentalists in interpreting structural biology experiments.


Standards for heart valve surgery in a 'Heart Valve Centre of Excellence'.
Surgical centres of excellence should include multidisciplinary teams with specialist expertise in imaging, clinical assessment and surgery for patients with heart valve disease. There should be structured training programmes for the staff involved in the periprocedural care of the patient and these should be overseen by national or international professional societies. Good results are usually associated with high individual and centre volumes, but this relationship is complex. Results of surgery should be published by centre and should include rates of residual regurgitation for mitral repairs and reoperation rates matched to the preoperative pathology and risk.


Mercury activates phospholipase a(2) and induces formation of arachidonic Acid metabolites in vascular endothelial cells.
ABSTRACT Currently, mercury has been identified as a risk factor in cardiovascular diseases among humans. Here, we tested our hypothesis that mercury modulates the activity of the vascular endothelial cell (EC) lipid signaling enzyme phospholipase A(2) (PLA(2)), which is an important player in the EC barrier functions. Monolayers of bovine pulmonary artery ECs (BPAECs) in culture, following labeling of membrane phospholipids with [(3)H]arachidonic acid (AA), were exposed to the inorganic form of mercury, mercury chloride, and the release of free AA (index of PLA(2) activity) and formation of AA metabolites were determined by liquid scintillation counting and enzyme immunoassay, respectively. Mercury chloride significantly activated PLA(2) in BPAECs in a dose-dependent (0 to 50 muM) and time-dependent (0 to 120 min) fashion. Metal chelators significantly attenuated mercury-induced PLA(2) activation, suggesting that cellular mercury-ligand interaction is required for the enzyme activation and that chelators are suitable blockers for mercury-induced PLA(2) activation in ECs. Sulfhydryl (thiol-protective) agents, calcium chelating agents, and cPLA(2)-specific inhibitor also significantly attenuated the mercury-induced PLA(2), suggesting the role of thiol and calcium in the activation of cPLA(2) in BPAECs. Significant formation of AA metabolites, including the release of total prostaglandins, thromboxane B(2), and 8-isoprostane, were observed in BPAECs following their exposure to mercury chloride. Mercury chloride induced cytotoxicity as observed by the altered cell morphology and enhanced trypan blue uptake, which was attenuated by the cPLA(2) inhibitor AACOCF(3). The results of this study revealed that inorganic mercury-induced PLA(2) activation through the thiol and calcium signaling and the formation of bioactive AA metabolites further demonstrated the association of PLA(2) with the cytotoxicity of mercury in ECs. Overall, the results of the current study underscore the importance of PLA(2) signaling in mercury-induced endothelial dysfunctions.


Agomelatine protects heart and aorta against lipopolysaccharide-induced cardiovascular toxicity via inhibition of NF-kβ phosphorylation.
The aim of this study was to investigate the possible ameliorating effects of agomelatine (AGO) on lipopolysaccharide (LPS)-induced endothelial and cardiac damage. Twenty-four female Wistar Albino rats divided into 3 groups as follows: Control, LPS and LPS + AGO. Total oxidant status (TOS), total antioxidant status (TAS), nuclear factor kappa beta (NF-kβ)/p65, p-NF-kβ, full caspase-8 (Cas-8) and cleaved cas-8 levels were measured in cardiac tissues and creatine kinase MB (CKMB), aspartate aminotransferase (AST), lactate dehydrogenase (LDH) levels in blood biochemically. In addition; cas-8, sirtuin-1 (SIRT-1), interleukin-4 (IL-4), interleukin-10 (IL-10), haptoglobin measured histopathologically in cardiac and aortic tissues. The levels of CKMB, AST, LDH and TOS were increased and TAS were decreased in the LPS group. In Western blot analyses NF-kβ/p65, p-NF-kβ/p65, full and cleaved cas-8 protein levels increased in cardiac tissues of LPS group. In histopathological and immunohistochemical evaluation of the heart sections; hyperemia, micro-hemorrhages and inflammatory cell infiltrations, increase of cas-8, haptoglobin, IL-4 and IL-10 and decrease of SIRT-1 levels were observed in cardiac and endothelial tissues of LPS groups. AGO treatment reversed all these parameters. It was shown that LPS-induced inflammation, oxidative stress and apoptosis via increasing of NF-kβ/p65 signaling, decreasing of SIRT-1 levels and increase of cas-8 levels in heart and endothelial tissues respectively. AGO corrected all these parameters by its antioxidant, antiinflammatory and antiapoptotic activities.


Mechanistic study of the oxidation of caffeic acid by digital simulation of cyclic voltammograms.
The oxidation mechanism of caffeic acid (CAF) has been studied by means of cyclic voltammetry with the plastic formed carbon or glassy carbon electrode. CAF gives a well-developed two-electron reversible wave in acidic media, whereas it shows an irreversible behavior, i.e., a decrease of the rereduction peak, in less acidic media, suggesting that the oxidation of CAF follows an irreversible chemical reaction(s). Digital simulation analyses based on different oxidation mechanisms have been performed for the voltammograms obtained with the GC electrode in 1:1 (v/v) water:ethanol solutions. The results clearly show that the seeming two-electron oxidation of CAF occurs stepwise via one-electron processes, each of which follows an irreversible chemical reaction. It has also been suggested that the semiquinone radical as an intermediate of the one-electron oxidation should play an important role in the oxidation reaction. Evaluations of the rate constants for the chemical reactions have further suggested that the chemical reactions are dimerization reactions.


MicroRNA-935 acts as a prognostic marker and promotes cell proliferation, migration, and invasion in colorectal cancer.
Colorectal cancer (CRC) is a common cause of cancer-related deaths worldwide. MicroRNA-935 (miR-935) has been reported to be involved in several cancers. In the present study, we aimed to investigate the role of miR-935 in the progression of CRC. The expression levels of miR-935 in CRC tissues and cells were detected using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The prognostic significance of miR-935 was identified using Kaplan-Meier survival analysis and Cox regression assay. Cell proliferation capacity was detected by Cell Counting Kit-8 (CCK-8) assay. Cell migration and invasion capacities were determined by Transwell assay in CRC cells. Bioinformatics analysis and luciferase reporter assays were used to confirm the direct target of miR-935. The expression of miR-935 was increased in CRC tissues and cells. Overexpression of miR-935 was significantly associated with lymph node metastasis and TNM stage. Overexpression of miR-935 in patients predicted shorter overall survival compared to low miR-935 expression. The expression of miR-935 was indicated to be an independent prognostic factor for CRC patients. In addition, overexpression of miR-935 in CRC cells promoted cell proliferation, migration, and invasion, whereas inhibition of miR-935 suppressed cell proliferation, migration, and invasion. Bioinformatics and luciferase assays revealed that INPP4A is a direct target of miR-935. Our findings suggest that miR-935 functions as an oncogene and promotes CRC progression. INPP4A is a potential target of miR-935. And miR-935 may represent a prognostic biomarker and potential therapeutic strategy for CRC treatment.


Chloroplast Biogenesis 65 : Enzymic Conversion of Protoporphyrin IX to Mg-Protoporphyrin IX in a Subplastidic Membrane Fraction of Cucumber Etiochloroplasts.
The preparation from Percoll-purified cucumber (Cucumis sativus)etiochloroplasts of a subplastidic membrane fraction that is capable of high rates of Mg insertion into protoporphyrin IX is described. The plastid stroma was inactive when used either alone or in combination with the membrane fraction. Successful preparation of the subplastidic membrane fraction required that Mg-protoporphyrin chelatase was first stabilized by its substrate. This was achieved by lysing Percoll-purified plastids in a fortified hypotonic medium containing protoporphyrin IX prior to ultracentrifugation and separation of the stroma from the plastid membranes. Protoporphyrin IX became membrane bound. Other additives needed for enzyme activity fell into two groups: (a) those needed for enzyme stabilization during membrane preparation and (b) those involved in the primary mechanism of Mg insertion into protoporphyrin IX. Ethylenediaminetetraacetate belonged to the first group, magnesium belonged to the second group, and ATP belonged to both groups.


Cell Death in the Lung: The Apoptosis-Necroptosis Axis.
Regulated cell death is a major mechanism to eliminate damaged, infected, or superfluous cells. Previously, apoptosis was thought to be the only regulated cell death mechanism; however, new modalities of caspase-independent regulated cell death have been identified, including necroptosis, pyroptosis, and autophagic cell death. As an understanding of the cellular mechanisms that mediate regulated cell death continues to grow, there is increasing evidence that these pathways are implicated in the pathogenesis of many pulmonary disorders. This review summarizes our understanding of regulated cell death as it pertains to the pathogenesis of chronic obstructive pulmonary disease, asthma, idiopathic pulmonary fibrosis, acute respiratory distress syndrome, and pulmonary arterial hypertension.


Host-parasite relationships in flatfish (Pleuronectiformes)--the relative importance of host biology, ecology and phylogeny.
The extent to which host biology, ecology and phylogeny determine the diversity of macroparasite assemblages has been investigated in recent years in several taxa, including fish. However, consensus has not been reached probably as a result of data being collected from different sources, different temporal scales or host and parasite biogeography and phylogeny having greater influence than expected. The present study evaluates the relative importance of 27 biological, ecological and phylogenetic characteristics of 14 flatfish species on the diversity of their ecto- and endoparasite assemblages, comprising a total of 53 taxa. Redundancy analyses were applied to the mean abundance of each parasite taxa infecting each host and to the richness, taxonomic distinctness and variance in taxonomic distinctness calculated for each assemblage within each host. Only a few host characteristics contributed significantly to the observed patterns: host distribution was more important in determining the type and mean abundance of ectoparasites present in an assemblage, whereas diversity of these assemblages were mainly related to the host's maximum size. Endoparasite mean abundance and diversity were mostly influenced by the number of food items ingested and by the presence of Crustacea and Polychaeta in the diet. However, the sympatric occurrence of related hosts also played an important role in the diversity values found in macroparasite assemblages. Results showed that a host characteristic has different importance according to the host-parasite relationship being examined, suggesting an important role for host-parasite co-evolution on the diversity of extant macroparasite assemblages.


The Roles of Floral Organ Genes in Regulating Rosaceae Fruit Development.
The function of floral organ identity genes, APETALA1/2/3, PISTILLATA, AGAMOUS, and SEPALLATA1/2/3, in flower development is highly conserved across angiosperms. Emerging evidence shows that these genes also play important roles in the development of the fruit that originates from floral organs following pollination and fertilization. However, their roles in fruit development may vary significantly between species depending on the floral organ types contributing to the fruit tissues. Fruits of the Rosaceae family develop from different floral organ types depending on the species, for example, peach fruit flesh develops from carpellary tissues, whereas apple and strawberry fruit flesh develop from extra-carpellary tissues, the hypanthium and receptacle, respectively. In this review, we summarize recent advances in understanding floral organ gene function in Rosaceae fruit development and analyze the similarities and diversities within this family as well as between Rosaceae and the model plant species Arabidopsis and tomato. We conclude by suggesting future research opportunities using genomics resources to rapidly dissect gene function in this family of perennial plants.


Prolonged stability and sustained prodrug cell killing activity using receptor-mediated delivery of malarial circumsporozoite-cytosine deaminase fusion protein into liver cancer cells.
An effective strategy of delivering recombinant DNA or protein by nonviral vectors faces two major challenges: (a) the selective delivery to the specific target tissue; and (b) a long-term expression of the protein once inside the cells. The present study describes a receptor-mediated delivery strategy using recombinant fusion protein consisting of malaria circumsporozoite (CS) protein as a ligand and bacterial cytosine deaminase (CD), which catalyzes the production of 5-fluorouracil from its prodrug 5-fluorocytosine. We demonstrate that the CD-CS fusion protein can be internalized in a receptor-mediated manner, providing a target delivery. The internalized CD-CS is capable of synthesizing 5-fluorouracil from the exogenously added 5-fluorocytosine and elicits cell killing with bystander activities. Most importantly, the internalized recombinant protein is stable and remains functional for at least several days, probably because of the entrapment of the fusion protein in particular cytoplasmic compartments that are free from cytoplasmic degradation machinery. Thus, it is possible to use a simple recombinant fusion strategy to enhance intracellular protein stability for manufacturing biological active product in a cell type-specific manner. The application of this strategy in the treatment of liver cancers and liver metastasis of colorectal cancers is discussed.


Dietary determinants of energy intake and weight regulation in healthy adults.
Until recently, the percentage of energy from dietary fat has been considered a primary determinant of body fatness. This review covers recent studies from our laboratory that challenge this notion. High and low fat diets matched for energy density, palatability and fiber resulted in similar mean voluntary energy intakes over 9 d; analysis of the individual foods in these diets showed that energy density and palatability were significant determinants of energy intake, independent of fat content. Path analysis further revealed that the influence of energy density on energy intake was in part direct, and in part indirect and mediated by palatability. In another study, dietary variety within food groups was shown to be an important predictor of body fatness, and the direction of the association depended on which food groups provided the variety, i.e., the variety of sweets, snacks, condiments, entrees and carbohydrates consumed was positively associated with body fatness, whereas the variety of vegetables was negatively associated. Last, a study of restaurant food and body fatness showed that the frequency of consumption of restaurant food was positively associated with body fatness, independent of education level, smoking status, alcohol intake and physical activity. Restaurant meals tend to be high in fat and low in fiber, and thus energy dense. Restaurants also typically serve a variety of palatable foods in large portions. The increasing variety of high energy foods available and the increasing proportion of household income spent on foods consumed away from home may help explain the U.S. national rising prevalence of obesity.


Minimally invasive approach for coronary artery fistula correction.
Minimally invasive cardiac surgery has recently become widespread. The aim of this study is to analyse the feasibility of a minimally invasive approach for coronary artery fistula (CAF) correction. From February 2001 to June 2014, 49 patients in our centre underwent minimally invasive CAF correction without cardiopulmonary bypass (CPB). The mean age was 21.18 ± 11.24 years (range, 5 months to 64 years). Patients with cardiac anomalies who underwent CAF correction and those who underwent CAF via the minimally invasive approach with CPB were excluded. There were no in-hospital mortalities or conversions to median sternotomy. Subxiphoid incisions (22 cases) and parasternal incisions (20 cases) were the most commonly used approaches for the procedure. The operative time was 67.45 ± 22.69 min (30-125 min). The intubation time was 3.72 ± 1.82 h (2-12 h), and the ICU stay was 9.67 ± 5.43 h (4-24 h). A trivial residual shunt was identified in only 1 patient during the procedure; however, this shunt had disappeared by discharge. The minimally invasive approach can provide excellent surgical exposure for CAF ligation in select patients. This approach is a safe and cosmetic alternative to conventional treatment and may minimize the length of stay.


[Review: New antimicrobial agent series XXII: Aspoxicillin].
The results of clinical and laboratory studies on aspoxicillin (ASPC) are summarized in this paper. 1. ASPC possesses a broad antibacterial spectrum in vitro against Gram-positive and Gram-negative bacteria. ASPC shows more potent activity in vivo and stronger bactericidal action than expected from in vitro activity. 2. Peak blood levels of ASPC after intravenous injection or intravenous drip infusion are dose dependent and half-lives of ASPC in these cases are about 1.6 hours. ASPC is excreted in active form mostly into urine via kidney. ASPC is satisfactorily transferred into various tissues and body fluids, such as bile, sputum. The binding rate of ASPC to serum protein is much lower than penicillin derivatives like piperacillin (PIPC), sulbenicillin (SBPC) and ampicillin (ABPC). 3. In an open clinical trial of ASPC, 1,845 cases were evaluated. Clinical effects were excellent in 543 cases (29.4%) and good in 822 cases (44.6%), and the efficacy rate for cases judged as excellent and good comprised 74.0%. 4. Comparative studies in which the efficacy of ASPC was compared to efficacies of PIPC and SBPC were performed in patients with respiratory tract infections, postoperative wound infections and suppurative otitis media. ASPC showed satisfactory clinical effects in all trials. 5. In the above open and comparative clinical studies of ASPC, incidence of adverse side reaction was only 1.95% (45/2,304), and main side effects were skin rash and diarrhea.


Natural Polymers for Organ 3D Bioprinting.
Three-dimensional (3D) bioprinting, known as a promising technology for bioartificial organ manufacturing, has provided unprecedented versatility to manipulate cells and other biomaterials with precise control their locations in space. Over the last decade, a number of 3D bioprinting technologies have been explored. Natural polymers have played a central role in supporting the cellular and biomolecular activities before, during and after the 3D bioprinting processes. These polymers have been widely used as effective cell-loading hydrogels for homogeneous/heterogeneous tissue/organ formation, hierarchical vascular/neural/lymphatic network construction, as well as multiple biological/biochemial/physiological/biomedical/pathological functionality realization. This review aims to cover recent progress in natural polymers for bioartificial organ 3D bioprinting. It is structured as introducing the important properties of 3D printable natural polymers, successful models of 3D tissue/organ construction and typical technologies for bioartificial organ 3D bioprinting.


An observational cohort study on pre-operative investigations and referrals: How far are we following recommendations?
Pre-operative investigations are often required to supplement information for risk stratification and assessing reserve for undergoing surgery. Although there are evidence-based recommendations for which investigations should be done, clinical practice varies. The present study aimed to assess the pre-operative investigations and referral practices and compare it with the standard guidelines. The present observational study was carried out during 2014-appen2015 in a teaching institute after the approval from Institute Ethical Committee. A designated anaesthesiologist collected data from the completed pre-anaesthetic check-up (PAC) sheets. Investigations already done, asked by anaesthesiologists as well as referral services sought were noted and compared with an adapted master table prepared from standard recommendations and guidelines. Data were expressed in frequencies, percentage and statistically analysed using INSTAT software (GraphPad Prism software Inc., La Zolla, USA). Seventy-five out of 352 patients (42.67% male, 57.33% female; American Society of Anesthesiologists physical status I to III) were included in this study. Nearly, all patients attended PAC with at least 5 investigations done. Of them, 89.33% were subjected to at least one unnecessary investigation and 91.67% of the referral services were not required which lead to 3.5 ( SD ±1.64) days loss. Anaesthesiologist-ordered testing was more focused than surgeons. More than two-third of pre-operative investigations and referral services are unnecessary. Anaesthesiologists are relatively more rational in ordering pre-operative tests yet; a lot can be done to rationalise the practice as well as reducing healthcare cost.


The top ten concerns about recovery encountered in mental health system transformation.
The notion of "recovery" has recently taken center stage in guiding mental health policy and practice. However, it is not yet clear what the term means and what is to be entailed in transforming the nation's mental health system to promote it. The authors discuss the various meanings of recovery as applied to mental illness and list the top ten concerns encountered in efforts to articulate and implement recovery-oriented care. These concerns include the following: recovery is old news, recovery-oriented care adds to the burden of already stretched providers, recovery involves cure, recovery happens to very few people, recovery represents an irresponsible fad, recovery happens only after and as a result of active treatment, recovery-oriented care is implemented only through the addition of new resources, recovery-oriented care is neither reimbursable nor evidence based, recovery-oriented care devalues the role of professional intervention, and recovery-oriented care increases providers' exposure to risk and liability. These concerns are addressed through discussion of the two over-arching challenges that they pose, namely the issues of resources and risk.


Atorvastatin restores imbalance of cluster of differentiation 4 (CD4)+ T cells in immune thrombocytopenia in vivo and in vitro.
Immune thrombocytopenia (ITP) is an autoimmune haemorrhagic disease, in which the overactivation of T cells is crucial in the pathogenesis. Atorvastatin (AT), a lipid-lowering medicine, has shown promising immunomodulatory effects in certain inflammatory conditions. However, the immunoregulatory role of AT in ITP remains elusive. To investigate the effect of AT in the treatment of ITP, cluster of differentiation 4 (CD4)+ T cells were isolated from patients with ITP and cultured with different dosages of AT. We found that AT significantly inhibited cell proliferation, led to cell cycle arrest, induced apoptosis, and repressed the activation of CD4+ T cells in vitro. ITP murine models were then established, and results showed that AT treatment led to faster recovery of the platelet count to normal and exhibited comparable immunomodulatory function. Furthermore, we found the phosphorylation of mammalian target of rapamycin (mTOR), protein kinase B (AKT) and extracellular signal-regulated kinase (ERK), as well as activation of rat sarcoma virus (RAS) were all reduced dramatically after AT treatment in vitro. In conclusion, our present study demonstrated that AT could reinstate the functions of CD4+ T cells by inhibiting the excessive activation, proliferation, and survival of CD4+ T cells in ITP via the RAS/mitogen-activated protein kinase kinase (MEK)/ERK and the mTOR/phosphatidylinositol-3 kinase (PI3K)/AKT pathway. Therefore, we propose that AT could be used as a potential therapeutic option for ITP by restoring the over-activated cellular immunity.


Study of heterogeneity of lectins in mistletoe preparations by monoclonal antibodies to their A-subunits.
Monoclonal antibodies (monAbs) displaying diverse affinity to native and recombinant mistletoe lectins A-chains (ML I-A, ML II-A, ML III-A and rML-A) have been obtained. In accordance to specificity of monAb they have been classified into three groups: 1. monAb against MLI-A and MLII-A, 2. monAb against MLII-A and MLIII-A, 3. monAb against A-subunits of MLI, MLII and MLIII. The results indicate, that antigen determinants of mistletoe lectins recognized by monAb MNA4, MNA9 and MTC12 do not contain any carbohydrates. Assay of lectins in mistletoe preparations was based on enzyme-linked lectin assay to meet the requirements given in the guidelines for drug tests. In this paper a sandwich ELISA test-system is described which allows to identify each of three ML-toxins. With the detection limits below 3 ng/ml and a linear measuring range of 3-30 ng/ml, dosages of mistletoe lectins in therapeutic range can be assayed. The problems in mistletoe lectins determination, structural differences and nature of heterogeneity of this proteins are discussed.


Recommendations for the Use of Automated Gray Matter Segmentation Tools: Evidence from Huntington's Disease.
The selection of an appropriate segmentation tool is a challenge facing any researcher aiming to measure gray matter (GM) volume. Many tools have been compared, yet there is currently no method that can be recommended above all others; in particular, there is a lack of validation in disease cohorts. This work utilizes a clinical dataset to conduct an extensive comparison of segmentation tools. Our results confirm that all tools have advantages and disadvantages, and we present a series of considerations that may be of use when selecting a GM segmentation method, rather than a ranking of these tools. Seven segmentation tools were compared using 3 T MRI data from 20 controls, 40 premanifest Huntington's disease (HD), and 40 early HD participants. Segmented volumes underwent detailed visual quality control. Reliability and repeatability of total, cortical, and lobular GM were investigated in repeated baseline scans. The relationship between each tool was also examined. Longitudinal within-group change over 3 years was assessed via generalized least squares regression to determine sensitivity of each tool to disease effects. Visual quality control and raw volumes highlighted large variability between tools, especially in occipital and temporal regions. Most tools showed reliable performance and the volumes were generally correlated. Results for longitudinal within-group change varied between tools, especially within lobular regions. These differences highlight the need for careful selection of segmentation methods in clinical neuroimaging studies. This guide acts as a primer aimed at the novice or non-technical imaging scientist providing recommendations for the selection of cohort-appropriate GM segmentation software.


Two (betaalpha)(8)-barrel enzymes of histidine and tryptophan biosynthesis have similar reaction mechanisms and common strategies for protecting their labile substrates.
The enzymes N'-[(5'-phosphoribosyl)formimino]-5-aminoimidazole-4-carboxamide ribonucleotide isomerase (HisA) and phosphoribosylanthranilate isomerase (TrpF) are sugar isomerases that are involved in histidine and tryptophan biosynthesis, respectively. Both enzymes have the (betaalpha)(8)-barrel fold and catalyze Amadori rearrangements of a thermolabile aminoaldose into the corresponding aminoketose. To identify those amino acids that are essential for catalysis, conserved residues at the active sites of both HisA and TrpF from the hyperthermophile Thermotoga maritima were replaced by site-directed mutagenesis, and the purified variants were investigated by steady-state enzyme kinetics. Aspartate 8, aspartate 127, and threonine 164 appeared to be important for the HisA reaction, whereas cysteine 7 and aspartate 126 appeared to be important for the TrpF reaction. On the basis of these results and the X-ray structure of a complex between TrpF and a bound product analogue, a reaction mechanism involving general acid-base catalysis and a Schiff base intermediate is proposed for both enzymes. A comparison of the HisA and TrpF enzymes from T. maritima and Escherichia coli showed that, at the physiological temperatures of 80 and 37 degrees C, respectively, the enzymes from the hyperthermophile have significantly higher catalytic efficiencies than the corresponding enzymes from mesophiles. These results suggest that HisA and TrpF have similar chemical reaction mechanisms and use the same strategy to prevent the loss of their thermolabile substrates.


Complete artificial saliva alters expression of proinflammatory cytokines in human dermal fibroblasts.
Complete artificial saliva (CAS) is a saliva substitute often used as a vehicle for test articles, including smokeless tobacco products. In the course of a study employing normal adult human dermal fibroblasts (HDFa) as a model in vitro, we discovered that CAS as a vehicle introduced a significant change in the expression of proinflammatory cytokines. To determine the effects of CAS on gene expression, real-time quantitative reverse-transcriptase PCR gene array analysis was used. Results indicate that robust changes in the expression of the proinflammatory cytokine interleukin 8 (IL8) and the vascular cell adhesion molecule 1 (VCAM1) occur within 5h of exposure to CAS. To determine whether CAS also alters cytokine release into the culture media, cytometric bead array assays for human inflammatory cytokines were performed. Analysis shows that CAS induced the release of IL8 and IL6. This study focused on determining which components in CAS were responsible for the proinflammatory response in HDFa. The following components were investigated: α-amylase, lysozyme, acid phosphatase, and urea. Results demonstrated that enzymatically active α-amylase induced gene expression for proinflammatory cytokines IL8, IL6, tumor necrosis factor-α, and IL1α and for VCAM1. Therefore, it is important to carefully evaluate the "vehicle effects" of CAS and its components in in vitro toxicology research.


Experimental femoral fracture immobilized by rigid and flexible rods (a rabbit model).
Experimental fractures in rabbits demonstrated that flexible intramedullary rod fixation of the midshaft long bone fracture enhances the fracture healing and may promote early mobility. When compared to rigid fixation, both excess flexibility and excess rigidity inhibited optimal fracture healing. The occurrence of angulation deformities with plastic rods also demonstrate the disadvantage of a very flexible rod.


[Trends and new factors affecting the brain drain from developing countries].
Current trends in brain-drain migration from developing to developed countries are analyzed. "Describing the growing specialisation of and rapid changes in the demand for qualified migrants in developed countries, this study shows that the brain drain phenomenon is increasingly fed by LDCs nationals migrating primarily for studies abroad. The resulting economic and social effects, both for sending and receiving countries, of the brain drains' modern patterns are analysed, as is the current action undertaken by various parties to eradicate this phenomenon." (SUMMARY IN ENG)


Role of the asialoglycoprotein receptor in binding and entry of hepatitis C virus structural proteins in cultured human hepatocytes.
We used a baculovirus-based system to prepare structural proteins of hepatitis C virus (HCV) genotype 1a. Binding of this preparation to cultured human hepatic cells was both dose dependent and saturable. This binding was decreased by calcium depletion and was partially prevented by ligands of the asialoglycoprotein receptor (ASGP-R), thyroglobulin, asialothyroglobulin, and antibody against a peptide in the carbohydrate recognition domain of ASGP-R but not preimmune antibody. Uptake by hepatocytes was observed with both radiolabeled and dye-labeled HCV structural proteins. With hepatocytes expressing the hH1 subunit of the ASGP-R fused to green fluorescent protein, we could show by confocal microscopy that dye stain cointernalized with the fusion protein in an area surrounding the nucleus. Internalization was more efficient with a preparation containing p7 than with one that did not. The two preparations bound to transfected 3T3-L1 cells expressing either both (hH1 and hH2) subunits of the ASGP-R (3T3-22Z cells) or both hH1 and a functionally defective variant of hH2 (3T3-24X cells) but not to parental cells. Additionally, uptake of dye-labeled preparation containing p7 was observed with 3T3-22Z cells but not with 3T3-L1 or 3T3-24X cells or with the preparation lacking p7, suggesting that p7 regulates the internalization properties of HCV structural proteins. Our observations suggest that HCV structural proteins bind to and cointernalize with the ASGP-R in cultured human hepatocytes.


MORPHOMETRIC SUBTYPING FOR A PANEL OF BREAST CANCER CELL LINES.
A panel of cell lines of diverse molecular background offers an improved model system for high-content screening, comparative analysis, and cell systems biology. A computational pipeline has been developed to collect images from cell-based assays, segment individual cells and colonies, represent segmented objects in a multidimensional space, and cluster them for identifying distinct subpopulations. While each segmentation strategy can vary for different imaging assays, representation and subpopulation analysis share a common thread. Application of this pipeline to a library of 41 breast cancer cell lines is demonstrated. These cell lines are grown in 2D and imaged through immunofluorescence microscopy. Subpopulations in this panel are identified and shown to correlate with previous subtyping literature that was derived from transcript data.


Guarding the frontiers: the biology of type III interferons.
Type III interferons (IFNs) or IFN-λs regulate a similar set of genes as type I IFNs, but whereas type I IFNs act globally, IFN-λs primarily target mucosal epithelial cells and protect them against the frequent viral attacks that are typical for barrier tissues. IFN-λs thereby help to maintain healthy mucosal surfaces through immune protection, without the significant immune-related pathogenic risk associated with type I IFN responses. Type III IFNs also target the human liver, with dual effects: they induce an antiviral state in hepatocytes, but specific IFN-λ4 action impairs the clearance of hepatitis C virus and could influence inflammatory responses. This constitutes a paradox that has yet to be resolved.


Rodent species as natural reservoirs of Borrelia burgdorferi sensu lato in different habitats of Ixodes ricinus in The Netherlands.
Rodents are natural reservoirs for human pathogenic spirochaetes of the Borrelia burgdorferi complex [B. burgdorferi sensu lato (s.l.)], and the pathogens are transmitted by Ixodes ricinus ticks to humans in The Netherlands. B. burgdorferi s.l. infection prevalence in questing ticks, rodents, and ticks feeding on these rodents, all sampled within the same short time span of five days in three different areas in The Netherlands, were compared in order to establish the relationship between ticks, reservoir hosts, and B. burgdorferi s.l. Questing nymphs were found in all 3 areas and numbers differed per area and even per site within areas. Infection prevalence in questing nymphs ranged between 0 and 20%. Apodemus sylvaticus and Myodes glareolus were the dominant rodents captured, and their numbers differed per area. Infection prevalence, determined by ear biopsies, ranged between 0 and 33.3% for both rodent species. Larvae were most frequently found feeding on these rodents, and their Borrelia infection prevalence ranged between 0 and 6.3% (A. sylvaticus) and between 0 and 29.4% (M. glareolus). The burden of nymphs feeding on rodents was low and varied per area with only 2 of 42 nymphs infected. Comparisons made on the basis of infection prevalence indicated that there was no clear relationship between rodents and questing nymphs when sampled within the same short time span. However, a possible relationship was present when questing ticks were sampled over longer periods in time (months) within or near the same areas (range of infection prevalence between 3.7 and 39.4). Confounding factors thus play a role in the interaction between rodents, ticks, and B. burgdorferi s.l., and it is very likely that other reservoir host species are responsible for the observed fluctuations. It is concluded that the local variations in rodent-Borrelia-tick interactions only partially explain the Lyme borreliosis risk in the sites studied and that other ecological determinants, notably vertebrate hosts and vegetation structure, should be incorporated in future studies of Lyme borreliosis risk.


Specific aspects of the treatment of patients with multiple mechanical and burn injuries.
Fewer than 5 percent of all burn patients sustain other traumatic injuries before, during, or immediately after their accident. Multiple traumas associated with a burn injury create special problems during the care of the thermally injured patients. Conversely, the burn injury often complicates the diagnosis and treatment of the trauma. The combination of mechanical and burn injuries can be divided into two types: a) any associated fracture located outside the burned area, or b) fractured bones within the burned area. This situation represents a critical factor which must be taken into account during treatment procedures. The following options should be considered: in fractures outside the burned area, there would be no difference in standard, skeletal treatment procedures. For the treatment of fractures in burned areas (mostly on the extremities), the optimal procedure is osteosynthesis within 48 hours of the burn trauma, when the burn wound is nearly sterile (without significant bacterial colonisation). We prefer two-team surgery. Firstly, a trauma surgeon performs osteosynthesis and, after that, burn surgeons treat the burns. The optimal approach in full-thickness burns would be necrectomy and autografting. This is, however, not always possible, because of the overall condition of the patient who has been continuously resuscitated during the shock period. All individual factors must be considered during the decision-making process.


Reduced prostaglandin release from the stunned porcine heart--significance for attenuated reactive hyperaemia.
Since both reactive hyperaemia and membrane phospholipids are altered even after short lasting ischaemic periods, the release of PGE2 and PGI2 in the basal state and during early reperfusion was examined to determine whether it was changed in the stunned myocardium. The effect of prostaglandin synthesis inhibition on reactive hyperaemia was also examined. The distal left anterior descending coronary artery was occluded for brief periods and coronary flow was recorded by Doppler flowmetry. In subgroups: (1) a shunt was established draining the ischaemic region for determination of myocardial prostaglandin release associated with 2 min of ischaemia before and after a 10 min occlusion; (2) prostaglandin synthesis was blocked between two 2 min occlusions by infusing indomethacin into the left anterior descending artery; and (3) segment lengths were measured in the left anterior descending artery region subjected to consecutive periods of 2, 10, and 2 min of ischaemia, and in a control region. 21 pentobarbitone sodium anaesthetised pigs, weight 21-30 kg, were used. 30 min after the 10 min occlusion, systolic shortening was reduced by 38(18-57)% (median +95% confidence interval; p less than 0.05). Concomitantly, basal PGE2 and PGI2 release was reduced by 69(30-77)% (p less than 0.05) and 58(7-81)% (p less than 0.05), respectively. During early reperfusion after 2 min of ischaemia, PGE2 release was reduced by 53(17-86)% (p less than 0.05) after development of stunning, whereas PGI2 release remained unaltered. Blockade of prostaglandin synthesis did not affect reactive hyperaemia either in normal or in stunned myocardium. Prostaglandin release from the stunned myocardium is reduced. Since indomethacin did not affect reactive hyperaemia, the attenuated PGE2 release during early reperfusion in stunned myocardium cannot explain the concomitant reduction in reactive hyperaemia.


Impacts of diabetes mellitus on clinical and para-clinical parameters among COVID-19 patients.
Diabetes has several adverse effects on patients with coronavirus disease 2019 (COVID-19); however, the determinants of this effect are still poorly understood. It is tried in current study to evaluate impacts of type 2 diabetes, with and without other comorbidities, on the clinical, para-clinical, and outcome parameters among COVID-19 patients. A case series was applied, which involved 406 COVID-19 patients admitted in the city of Shiraz, south-central Iran, from February 20 to April 29, 2020. Demographic data, medical history, laboratory finding, chest computed tomography (CT) scan reports, and clinical outcomes of patients with and without type 2 diabetes were compared. Results of the above-mentioned comparison showed that comorbidities such as HTN (35.5% vs. 13.7%, p < 0.001) and CVDs (26.2% vs. 13.4%, P = 0.002) were significantly more prevalent among the diabetic patients. Also, there was not any considerable difference between the chest CT severity parameters of both groups. After excluding all of the comorbidities except diabetes, it was found that the diabetic COVID-19 patients without other comorbidities had lower oxygen saturation level (P < 0.001), higher AST level (P = 0.037), higher BUN (P = 0.005), higher WBC counts (P = 0.025), lower lymphocyte counts (P = 0.029), and longer ICU admission duration (0.72 ± 2.83 vs. 1.71 ± 4.68, P = 0.046). The diabetic COVID patients are at higher risks of hypoxemia, longer ICU stays, and more renal and hepatic dysfunction. These achievements could be useful in order to prevent the deterioration of clinical conditions among diabetic COVID-19 patients; also, they have to be considered in the management strategies.


A 30-year-old male with COVID-19 presenting with seizures and leukoencephalopathy.
First reported in Wuhan, China, Novel Coronavirus Disease-19 rapidly spread causing an outbreak of viral pneumonia and became a pandemic in early 2020. It was later discovered to be caused by Severe Acute Respiratory Syndrome Coronavirus 2, a novel coronavirus. Although the vast majority of cases have primarily involved the respiratory system, some serious cases have started to emerge with central and peripheral nervous system complications. We present the case of a 30-year-old morbidly obese male who initially presented to the emergency department with seizures, altered mental status, and COVID-19 pneumonia. After a 21-day hospital course including 14 days of intensive care unit management, he was stabilized and discharged to a rehabilitation facility. He returned 1 day later with worsening respiratory distress and was found to have acute pulmonary embolism requiring placement of an inferior vena cava filter. After an additional 6 days in the hospital, he was discharged back to the outpatient facility. He returned for a third time with altered mental status, visual and auditory hallucinations, and confabulation. This report provides critical information in revealing a peculiar neurological sequela of COVID-19 induced leukoencephalopathy and its disease course. We hope to shed light on this sequence of events by providing possible mechanisms to aid clinicians in the identification and management of this complication.


Localization of a promoter in the putative internal ribosome entry site of the Saccharomyces cerevisiae TIF4631 gene.
The 5'-region of the TIF4631 gene of Saccharomyces cerevisiae (encoding the translation initiation factor eIF4G1) was reported earlier to harbor a very active internal ribosome entry site (IRES) allowing for internal initiation of translation of TIF4631 mRNA. Here, we report the presence of a promoter in the region -112 to -36 relative to the translation initiation codon of the TIF4631 gene. This promoter stimulates transcription from a start site at position -36 and generates an mRNA that is actively translated in vitro and able to sustain growth of yeast cells in vivo as the only source of eIF4G. The data show that the IRES activity reported earlier is due to this promoter. On the contrary, the presumed IRES represents a strongly inhibitory element for translation in vitro.


On-line access to nursing literature.
Providing access to the multiple databases essential to nursing practice and education is a challenge for universities and health care providers alike. Collaboration between Wayne State University and other institutions in the Detroit metropolitan area provides a unique system for user access to nursing information. Building on the Detroit Area Library Network (DALNET) structure, additional computer databases have been added to the Library User Information System (LUIS) to allow faculty and student access from remote locations. With university identification, faculty and students access up-to-date literature-searching tools and check the availability of desired journal articles at multiple locations throughout the large tri-county area.


Quality assurance and reproducibility of high-resolution two-dimensional electrophoresis and silver staining in polyacrylamide gels.
Qualitative data from high-resolution two-dimensional electrophoresis (2DE) have become clinically useful in immunoglobulin clonality analysis, in resolution of ambiguities in immunofixation typing of paraproteins, and in genetic typing of serum proteins. Since 1986, the authors have been evaluating the College of American Pathologists Reference Preparation for Serum Proteins (RPSP) as a quality control material for 2DE because (1) it is prepared exclusively from pooled human sera, (2) the pool yields a reference pattern of mixed heterozygosity for genetic markers, and (3) RPSP is widely available as a lyophilized preparation that currently serves in the authors' laboratory as a qualitative quality control preparation and that may become a quantitative quality control material or external quality assessment material for 2DE. Using the ISO-DALT 2DE system and silver-staining, the peptide patterns were examined in 11 lots of RPSP and compared with fresh serum and with each other. Consistent differences in the 2DE pattern between RPSP and fresh serum included the presence of freeze-thaw peptides, the presence of degradation spots of apolipoprotein A-I, and the diminution of apolipoprotein spot intensities in RPSP. All lots of RPSP yielded clear identification of the eight serum proteins used for quality control calculations. Run-to-run coefficients of variation for a single lot of RPSP for four parameters of 2DE spot location and gradient reproducibility were comparable with band location reproducibility for the one-dimensional procedures of serum protein electrophoresis and lactate dehydrogenase isoenzyme electrophoresis. It is concluded that the reproducibility, that is, imprecision, of 2DE is the same as one-dimensional clinical electrophoresis techniques and that either RPSP or pooled fresh serum can serve as a satisfactory internal quality control material.


Long-term survival and sprouting in culture by motoneurons isolated from the spinal cord of adult frogs.
Motoneurons of adult frog spinal cord have been retrogradely labeled with the carbocyanine derivative diI. Spinal cords were then dissociated and the labeled motoneurons partially purified by centrifugation over a bovine serum albumin (BSA) density gradient. The resulting cell suspension was plated on a substrate of innervated muscle extracellular matrix (ECM) to which the motoneurons attached and extended processes. Labeled adult motoneurons survived for more than 4 weeks in a defined medium in the absence of added serum or growth factors. These cultures of adult motoneurons provide a favorable preparation for studying molecular factors that influence process outgrowth and synapse formation.


Docetaxel as salvage therapy in highly pretreated and drug resistant gastrointestinal carcinomas.
Despite many efforts to develop new chemotherapies for metastatic upper gastrointestinal (GI) cancer, overall prognosis continues to be fatal, particularly in gastric and pancreatic cancer. Many of these patients deserve second-or third-line treatment after failure of first-line chemotherapy. Therefore, we analysed toxicity and response rate of weekly docetaxel after failed upfront regimes in these upper GI cancer patients. Between 2001 and 2006, 18 patients received docetaxel based regimes (35 mg/m(2) weekly) after informed consent. Response rates were determined using RECIST criteria or tumor progression if clinically evident. Toxicities were graded based on NCI CTC criteria (version 2). Most patients had gastric cancer (13/18). The remaining entities comprised of bilio-pancreatic cancer (5/18). Docetaxel was administered as 2nd line therapy in 28% (5/18), 3rd line therapy in 56% (10/18) and 4th or 5th line therapy in 17% (3/18). The average docetaxel dose was 38 mg/m(2) (Median: 35 mg/m(2)) once weekly. Over a treatment duration of 14.7 weeks, the average dosage was 58 gr per patient and week. Overall, docetaxel was well tolerated with only few chemotherapy-associated toxicities (Grade 3/4), including nausea (17%), polyneuropathy (17%), anorexia (11%), neutropenia (6%) and leukopenia (17%). Docetaxel administration did not achieve any complete responses (CR) and one (5.6%) partial response (PR) was seen (1/18). In addition 5 patients (27.8%) had stable disease (SD), thus inducing a tumor control rate of 33.3% (6/18). Median progression-free survival was 2.4 months for all patients, 2.1 months in the gastric-cancer and 2.4 months in the bilio-pancreatic cancer subgroups respectively. After first docetaxel administration median survival for all patients was 4.5 months, patients with gastric cancer survived for 4.9 months whereas patients suffering from bilio-pancreatic carcinoma survived for 4.2 months. However, taken together 27% (5/18) had a remarkable overall survival of more than 2.5 years. In severely pretreated patients, with documented chemoresistant GI tumors, weekly docetaxel was well tolerated, presented good tumor control rate and overall survival. Therefore, this regimen may be used as salvage treatment in individual patients with upper GI cancers.


[A review of current studies on leisure activities performed by community-dwelling elders].
This article reviewed and summarized the findings of previously published studies on the leisure activities of community elders over 55 years of age. A total of 17 related articles, published before 2008, were identified in the search of electronic databases, which included the Chinese Periodical Table of Contents Index Image System, CEPS, Medline and Ageline. Leisure activities measurements used in this study consisted of recoding activities either categorically or by frequency. The majority of research indicated that elderly engage in primarily static activities and that participation in leisure activities had positive effects on cognitive and psychological health. However, inconsistencies in results were found due to differences in measurement tools and sample populations used among studies, making it difficult to conduct a valid comparison of specific features of elderly leisure activities. Most current leisure activity studies utilize cross-sectional research methodologies with restricted topics, which result in limited applicability of research results. In conclusion, we suggest that future studies should develop instruments to assess leisure activities that correspond to local conditions identified through qualitative research. With such, researchers can study different age groups by applying relevant ageing theories to construct empirical data related to current elderly leisure activities.


Cloning and characterization of the arginine-specific carbamoyl-phosphate synthetase from Bacillus stearothermophilus.
Bacillus stearothermophilus contains two carbamoyl-phosphate synthetases (CPS), one specific for pyrimidine biosynthesis and the other for arginine biosynthesis. The pyrimidine-specific CPS is repressed by exogenous pyrimidines, and its activity is inhibited by UMP and activated by 5-phospho-alpha-D-ribosyl diphosphate. The arginine-specific CPS is similarly repressed by exogenous arginine but its activity is not sensitive to these or other potential effectors. Each of the two enzymes consist of two unequal subunits, as is the case for other microbial CPS; however, the large subunit for the arginine-specific CPS is smaller than that for the pyrimidine-specific enzyme. Comparison of the derived amino acid sequence for the cloned large subunit of the arginine-specific CPS with those for subunits from pyrimidine-sensitive CPS showed significant similarity throughout the polypeptides except at the carboxy terminus, which was identified by other laboratories to contain the binding site for the pyrimidine effector. Unlike the results previously reported for CPS from an enteric mesophile, the kinetic properties of the arginine-specific CPS were not affected by growth of B. stearothermophilus at temperatures near the minimal growth temperature. Furthermore, calorimetric studies showed that the thermal stability of cloned CPS was identical regardless of the growth temperature of B. stearothermophilus between 42 degrees C and 63 degrees C. The thermal stability of cloned CPS was not affected by expression at 37 C in Bacillus subtilis or Escherichia coli. In contrast, the thermal stabilities for CPS and other proteins were higher in extracts of cells grown at higher temperatures. These results indicate that cellular factors, probably chaperonins, are necessary for thermal stability of proteins at and below the optimal temperature for this thermophile.


Analysis of Toxic Amyloid Fibril Interactions at Natively Derived Membranes by Ellipsometry.
There is an ongoing debate regarding the culprits of cytotoxicity associated with amyloid disorders. Although small pre-fibrillar amyloid oligomers have been implicated as the primary toxic species, the fibrillar amyloid material itself can also induce cytotoxicity. To investigate membrane disruption and cytotoxic effects associated with intact and fragmented fibrils, the novel in situ spectroscopic technique of Total Internal Reflection Ellipsometry (TIRE) was used. Fibril lipid interactions were monitored using natively derived whole cell membranes as a model of the in vivo environment. We show that fragmented fibrils have an increased ability to disrupt these natively derived membranes by causing a loss of material from the deposited surface when compared with unfragmented fibrils. This effect was corroborated by observations of membrane disruption in live cells, and by dye release assay using synthetic liposomes. Through these studies we demonstrate the use of TIRE for the analysis of protein-lipid interactions on natively derived lipid surfaces, and provide an explanation on how amyloid fibrils can cause a toxic gain of function, while entangled amyloid plaques exert minimal biological activity.


Use of the laryngeal mask airway during repair of atrial septal defect in children.
We describe the elective use of the laryngeal mask airway in two children undergoing cardiopulmonary bypass for repair of an atrial septal defect. Total surgical time was short and cardiopulmonary bypass performed at normothermia allowing removal of the laryngeal mask airway on the operating table on completion of surgery. We were able to adequately oxygenate and ventilate the children throughout the procedure using positive pressure ventilation and spontaneous ventilation. The use of caudal fentanyl and rectal diclofenac aided postoperative pain management. Atrial septal defect repair has become one of the more straightforward cardiac operations partly as a result of new cardiopulmonary bypass techniques. Avoidance of intubation and postoperative ventilation in appropriate patients would make this procedure ideal for 'fast tracking' and offers the potential advantages of decreased length of stay in hospital and reduction in overall costs.


Simulating microinjection experiments in a novel model of the rat sleep-wake regulatory network.
This study presents a novel mathematical modeling framework that is uniquely suited to investigating the structure and dynamics of the sleep-wake regulatory network in the brain stem and hypothalamus. It is based on a population firing rate model formalism that is modified to explicitly include concentration levels of neurotransmitters released to postsynaptic populations. Using this framework, interactions among primary brain stem and hypothalamic neuronal nuclei involved in rat sleep-wake regulation are modeled. The model network captures realistic rat polyphasic sleep-wake behavior consisting of wake, rapid eye movement (REM) sleep, and non-REM (NREM) sleep states. Network dynamics include a cyclic pattern of NREM sleep, REM sleep, and wake states that is disrupted by simulated variability of neurotransmitter release and external noise to the network. Explicit modeling of neurotransmitter concentrations allows for simulations of microinjections of neurotransmitter agonists and antagonists into a key wake-promoting population, the locus coeruleus (LC). Effects of these simulated microinjections on sleep-wake states are tracked and compared with experimental observations. Agonist/antagonist pairs, which are presumed to have opposing effects on LC activity, do not generally induce opposing effects on sleep-wake patterning because of multiple mechanisms for LC activation in the network. Also, different agents, which are presumed to have parallel effects on LC activity, do not induce parallel effects on sleep-wake patterning because of differences in the state dependence or independence of agonist and antagonist action. These simulation results highlight the utility of formal mathematical modeling for constraining conceptual models of the sleep-wake regulatory network.


Diagnosis of malaria in the febrile traveler.
To determine if clinical presentation can be used for predicting malaria infection in febrile patients with recent travel to a malaria-endemic area, 157 patients with malaria, a history of fever, and recent travel to a malaria-endemic area were compared with 157 matched controls in case-control study. Fever pattern, symptom duration, temperature, splenomegaly, and platelet count were correlated with malaria infection. These criteria, however, either singly or in combination, had low sensitivity for accurately identifying patients with malaria. Because no criterion could accurately predict the presence of malaria, it is concluded that microscopic examinations of blood for malaria parasites should be done in all symptomatic patients with a history of travel to a malaria-endemic area.


[Complications after cardiac injuries].
An analysis of 293 victims with cardiac and pericardium injuries was made. Cardiac complications could appear suddenly and be registered on any stage of treatment process. They differentiated by character and duration of wound process. Pericarditis took place in 288 patients. The syndrome of system inflammation reaction was noted in 47.9% of patients, a sepsis (sometimes severe sepsis) was in 14.3%. Complications in postoperative period were determined by posthypoxic and hemic hypoxia, coagulopathy. They were manifested by myocardial ischemia and thromboembolic complications. An acute myocardial infarction took place in 29 (9.8%) cases and rhythm and conductivity abnormalities were in 23 (7.8%) patients. The circulatory failure with clinical picture of pulmonary edema was developed in 12 (4.1%) cases. The lethality consisted of 44 (15%) patients.


[A case of tympanic foreign body].
A 22 years old male was hospitalized with 1 month history of endaural foreign body retention at 7th May, 2011. Patient complained an invicible bug was placed into right outer ear canal. The foreign body could not be taken out because of the intoleration to pain under the local anesthesia, and major symptoms were swell-sore, fullness, and obvious hearing loss. Physical examination showed congestion and swelling in right ear canal, and only 3mm diameter for the stenosis ear canal. The incrustation make the invicible foreign body and the tympanic membrane. CT showed the foreign body retention near tympanic membrane left ear was normal. After hospitalization, the patient received an exploratory operation under the general anesthesia. The foreign body incarceration was found near the tympanic ring which in the tympanic cavity.


Immunoglobulins in the cerebrospinal fluid of patients with late syphilis.
The concentrations of immunoglobulins G, A, and M were measured in 62 samples of cerebrospinal fluid from 35 patients with asymptomatic neurosyphilis, 18 with symptomatic neurosyphilis, and nine with late syphilis of other organs. The most frequent and highest increase in IgG and IgA occurred in patients with symptomatic neurosyphilis. IgM was found in only five patients, but its presence, together with an increase in IgG and IgA concentrations, appears to indicate activity of the pathological process and a poor prognosis for the course of the disease.


[Vacuum mixing systems for bone cement completion - comparison of different systems].
The use of vacuum mixing systems when mixing bone cement reduces its porosity and hereby significantly improves the features of the material. The currently available mixing systems are compared with regard to handling, mechanical properties and economical aspects. In 8 vacuum mixing systems the handling, pump performance, system tightness, the used air volume, the filter efficiency, the remaining amounts in the mixing system and the porosity of the cements are shown in comparison. All vacuum mixing systems reduce the porosity of the cement in comparison to hand mixed cements significantly if used correctly. The individual examinations, though, show enormous differences, which can be of significance to the user in the individual choice of system and, depending on the individual circumstances, can influence the quality of the mixed cement. The results enable the user to choose and handle a vacuum mixing system which is optimally suitable for the individual circumstances in respect to the characteristics examined.


The Petunia CHANEL Gene is a ZEITLUPE Ortholog Coordinating Growth and Scent Profiles.
The floral perianth, comprising sepals and petals, conceals the sexual organs and attracts pollinators. The coordination of growth and scent emission is not fully understood. We have analyzed the effect of knocking down CHANEL (PhCHL), the ZEITLUPE ortholog in petunia (PhCHL) by hairpin RNAs. Plants with low PhCHL mRNA had overall decreased size. Growth evaluation using time lapse image analysis showed that early leaf movement was not affected by RNAi:PhCHL, but flower angle movement was modified, moving earlier during the day in knockdown plants than in wild types. Despite differences in stem length, growth rate was not significantly affected by loss of PhCHL. In contrast, petal growth displayed lower growth rate in RNAi:PhCHL. Decreased levels of PhCHL caused strongly modified scent profiles, including changes in composition and timing of emission resulting in volatile profiles highly divergent from the wild type. Our results show a role of PhCHL in controlling growth and development of vegetative and reproductive organs in petunia. The different effects of PhCHL on organ development indicate an organ-specific interpretation of the down regulation of PhCHL. Through the control of both timing and quantitative volatile emissions, PhCHL appears to be a major coordinator of scent profiles.


Fundic gland polyps developing during omeprazole therapy.
Side effects of omeprazole therapy include reversible hypergastrinemia, entero chromaffin-like hyperplasia, and possibly acceleration of atrophic gastritis in the gastric corpus in patients with Helicobacter pylori infection. The objective of this report is to describe six cases of fundic gland polyps that developed while omeprazole was being used for Barrett's esophagus. Three women and three men developed fundic gland polyps after 1-5 yr of omeprazole therapy, 20 mg/day. In the index case, fundal polyps developed after 2 yr of continuous omeprazole therapy. Omeprazole was stopped, but because of poor clinical efficacy with H2-blockers, it was reinstated. After 4 yr of therapy, multiple large fundal polyps were present. Histology of all lesions revealed characteristic fundal gland polyps with multiple cystic dilations. None of the patients had H. pylori infection. Serum gastrin was normal in four and slightly increased in two. These cases support the possible causal relation between the use of omeprazole and the development of fundic gland polyps in patients without H. pylori gastritis.


Inhibition of rapamycin-induced autophagy causes necrotic cell death associated with Bax/Bad mitochondrial translocation.
Rapamycin, a lipophilic macrolide antibiotic, has been found to reduce injury in different models of neurodegenerative disorders. We have previously shown that in neonatal rats subjected to hypoxia-ischemia (HI) the neuroprotective effect of rapamycin was associated with increased autophagy and decreased caspase-3 activation. We show here that the strong reduction of caspase-3 activation after rapamycin was due, at least in part, to its effect on the intrinsic apoptotic mitochondrial pathway because after rapamycin treatment there was a marked reduction of Bax and Bad translocation to mitochondria, cytochrome c release, and caspase-3 activation. Poly (ADP-ribose) polymerase 1 (PARP-1) cleavage and the number of terminal dUDP nick-end labeling (TUNEL)-positive cells were also reduced. To assess how the antiapoptotic effect of rapamycin was linked to the strong autophagy signal induced by the drug, we blocked the formation of autophagosomes with 3-methyladenine (3MA). 3MA administered 10 min after rapamycin, elicited again Bax and Bad translocation to the mitochondria but did not cause cytochrome c release and caspase-3 activation. After 3MA treatment, cells underwent necrotic cell death. These data indicate that rapamycin administered before HI prevents the apoptotic signaling taking place through the mitochondrial pathway. We hypothesize that rapamycin confers a preconditioning-like protection and suggest that caution is necessary before using pharmacological agents targeting autophagy in neuroprotection because they could interfere with endogenous protective mechanisms.


The inhibitory effect of heparin and related glycosaminoglycans on neutrophil chemotaxis.
Clinical observations have shown that heparin has antiinflammatory activities. The effect of heparin on neutrophil chemotaxis was evaluated in vitro in the Boyden Chamber. This method enabled differentiation between the direct effects of heparin on neutrophil migration and locomotion, and its effects on chemotactic factors. Heparin inhibited both the random migration and directed locomotion of human neutrophils toward zymosan-activated serum (ZAS) and F-met-leu-phe (FMLP). Inhibition was found to be dependent on the concentrations of the heparin and of the chemotactic factors. No specific binding of heparin to the neutrophils could be demonstrated, and heparin's inhibitory effects were eliminated by simple washing of the cells. When added directly to the chamber containing chemotactic factor, heparin inhibited the chemotactic activity of ZAS but not that of FMLP, suggesting a direct inhibitory effect against C5a, the principal chemotactic factor in ZAS. Experiments performed with low-molecular-weight heparin, N-desulfated heparin, dextran sulfate, chondroitin sulfate and dextran indicated that the inhibitory effects of heparin on neutrophil chemotaxis are not related to its anticoagulant activity, but probably depend on the degree of sulfation of the heparin molecule.


Baicalin Ameliorates Pancreatic Fibrosis by Inhibiting the Activation of Pancreatic Stellate Cells in Mice with Chronic Pancreatitis.
Pancreatic inflammation and fibrosis are typical pathological features in chronic pancreatitis (CP). Activated pancreatic stellate cells (PSCs) have been regarded as the core event in the development of pancreatic fibrosis and are considered to be the key target for treatment of CP. Baicalin (C21H18O11), the main chemical composition of Baikal skullcap in the traditional Chinese medicines Dachaihu decoction (DCHD) and Xiaochaihu decoction (XCHD), has shown significant effects in the treatment of pancreatic fibrosis in CP mice; however, whether baicalin can inhibit the activation of PSCs and its underlying mechanism remain unclear. In this study, the influence of baicalin on activated PSCs in vitro and in vivo was investigated, and the results showed that Baicalin could significantly ameliorate the degree of pancreatic inflammation and fibrosis, while decreasing the levels of alpha-smooth muscle actin (α-SMA), F4/80 (surface markers of mouse macrophages), nuclear factor kappa-B (NF-κB), monocyte chemotactic protein 1 (MCP-1), and collagen type I alpha 1 (COL1A1)in the pancreas. Moreover, NF-κB and α-SMA were co-expressed in the pancreas of CP mice. Baicalin treatment markedly reduced the expression of co-location of α-SMA and NF-κB. In vitro, the protein expression levels of transforming growth factor-β receptor 1 (TGF-βR1), phosphorylated TGF-β activated kinase 1 p-TAK 1, and NF-κBp65 in PSCs were all remarkably reduced after treatment with baicalin. In addition, baicalin could inhibit MCP-1 mRNA expression in supernatant of activated PSCs, as well as the excessive migration of macrophages. Taken together, our findings indicated that baicalin could inhibit the TGF-β1/TGF-βR1/TAK1/NF-κB signaling pathway of activated PSCs, reduce the secretion of MCP-1, and further decrease the infiltration of macrophages and inflammation cells of the local microenvironment of the pancreas. Thus, this study provides a reliable experimental basis for baicalin in the prevention and treatment of CP.


Characterization of long term hormonal cell cultures from the MtT/w15 prolactin- and growth hormone-secreting rat pituitary tumor.
PRL-secreting epithelial cell lines (PECL) derived from Furth's MtT/W15 transplantable rat pituitary tumor have been studied as monolayer cultures for up to 18 months. Tumor epithelial cells were mechanically dispersed, plated at high density, and selectively enriched by picking and pooling colonies, thereby avoiding single cell-cloning conditions, in order to study the long term natural evolution of tumor cell populations in vitro. Fibroblast-like cells, prevalent in primary and early passages, underwent a series of morphological changes, which have been interpreted by others as in vitro aging or differentiation, and were eliminated from the cultures within 9 months. PECL phenotypes were heterogeneous and differed from sc MtT/W15 tumors with respect to morphological differentiation and modal chromosome number (48 for solid tumors: 48-92 for PECL; 42 for diploid). PECL were devoid of secretory granules by electron microscopy, but accumulated neutral lipid, complex lysosomes, and endocytotic vesicles. PECL differed from one another in plating efficiency (0.6-7.3%) and basal secretion rates for PRL and GH. Population doubling times decreased during the culture interval from 100 to 24-37 h in different PECL. RIA of medium from early cultures (less than 7 months) disclosed that PRL always exceeded GH levels, mimicking the serum hormone ratio of MtT/W15 tumor-bearing Wistar-Furth rats. Basal hormone release declined with time from a maximal initial rate of 93 micrograms/mg cell protein.24 h for PRL and 2.3 micrograms/mg cell protein.24 h for GH. GH production ceased by 7 months in all PECL, but PRL secretion persisted for 18 months. PRL production increased during exponential growth as a direct function of cell number and protein, and reached a maximum during confluency. Immunoperoxidase-conjugated antibody staining localized PRL in the Golgi region but not in secretory granules, consistent with a rapid release rate for the hormone. In preliminary experiments, rat hypothalamic extract (one hypothalamic extract RP-1 equivalent per dish) and 17 beta-estradiol (1 X 10(-8) M), but not TRH (5 X 10(-8) M), stimulated PRL secretion from PECL/F3, whereas only hypothalamic extract stimulated total cell protein after exposure to the agents for 3-6 days. Long term cultures of PECL exhibited reduced oncogenicity when transplanted in W/Fu rats. During serial passages, in vivo, PRL secretion decreased from 154 microgram/ml serum to the normal range within the same time framework that PRL secretion from cultured PECL/F3 was observed to decrease and stop in vitro. We believe that this phenomenon represents an expressed program within the cells rather than being merely a reflection of the in vitro environment.


A Retrospective Cohort Study on Mortality and Neurodevelopmental Outcomes of Preterm Very Low Birth Weight Infants Born to Mothers with Hypertensive Disorders of Pregnancy.
We examined the effects of maternal hypertensive disorders of pregnancy (HDP) on the mortality and neurodevelopmental outcomes in preterm very low birth weight (VLBW) infants (BW ≤1,500 g) based on their intrauterine growth status and gestational age (GA). We included singleton VLBW infants born at <32 weeks' gestation registered in the Neonatal Research Network Japan database. The composite outcomes including death, cerebral palsy (CP), and developmental delay (DD) at 3 years of age were retrospectively compared among three groups: appropriate for GA (AGA) infants of mothers with and without HDP (H-AGA and N-AGA) and small for GA (SGA) infants of mothers with HDP (H-SGA). The adjusted odds ratios (AOR) and 95% confidence intervals (CI) stratified by the groups of every two gestational weeks were calculated after adjusting for the center, year of birth, sex, maternal age, maternal diabetes, antenatal steroid use, clinical chorioamnionitis, premature rupture of membranes, non-life-threatening congenital anomalies, and GA. Of 19,323 eligible infants, outcomes were evaluated in 10,192 infants: 683 were H-AGA, 1,719 were H-SGA, and 7,790 were N-AGA. Between H-AGA and N-AGA, no significant difference was observed in the risk for death, CP, or DD in any GA groups. H-AGA had a lower risk for death, CP, or DD than H-SGA in the 24 to 25 weeks group (AOR: 0.434, 95% CI: 0.202-0.930). The odds for death, CP, or DD of H-SGA against N-AGA were found to be higher in the 24 to 25 weeks (AOR: 2.558, 95% CI: 1.558-3.272) and 26 to 27 weeks (AOR: 1.898, 95% CI: 1.427-2.526) groups, but lower in the 30 to 31 weeks group (AOR: 0.518, 95% CI: 0.335-0.800). There was a lack of follow-up data; however, the outcomes of liveborn preterm VLBW infants of mothers with HDP depended on their intrauterine growth status and GA at birth. · The effects of HDP on preterm low birth weight infants need to be further examined.. · The outcomes were not different between AGA infants with and without maternal HDP.. · The outcomes of SGA infants with maternal HDP were dependent on their GA..


Broadband adiabatic inversion cross-polarization to integer-spin nuclei with application to deuterium NMR.
Solid-state NMR (SSNMR) spectroscopy of integer-spin quadrupolar nuclei is important for the molecular-level characterization of a variety of materials and biological solids; of the integer spins, 2 H (S = 1) is by far the most widely studied, due to its usefulness in probing dynamical motions. SSNMR spectra of integer-spin nuclei often feature very broad powder patterns that arise largely from the effects of the first-order quadrupolar interaction; as such, the acquisition of high-quality spectra continues to remain a challenge. The broadband adiabatic inversion cross-polarization (BRAIN-CP) pulse sequence, which is capable of cross-polarization (CP) enhancement over large bandwidths, has found success for the acquisition of SSNMR spectra of integer-spin nuclei, including 14 N (S = 1), especially when coupled with Carr-Purcell/Meiboom-Gill pulse sequences featuring frequency-swept WURST pulses (WURST-CPMG) for T2 -based signal enhancement. However, to date, there has not been a systematic investigation of the spin dynamics underlying BRAIN-CP, nor any concrete theoretical models to aid in its parameterization for applications to integer-spin nuclei. In addition, the BRAIN-CP/WURST-CPMG scheme has not been demonstrated for generalized application to wideline or ultra-wideline (UW) 2 H SSNMR. Herein, we provide a theoretical description of the BRAIN-CP pulse sequence for spin-1/2 → spin-1 CP under static conditions, featuring a set of analytical equations describing Hartmann-Hahn matching conditions and numerical simulations that elucidate a CP mechanism involving polarization transfer, coherence exchange, and adiabatic inversion. Several experimental examples are presented for comparison with theoretical models and previously developed integer-spin CP methods, demonstrating rapid acquisition of 2 H NMR spectra from efficient broadband CP.


Collaborative Multifidelity-Based Surrogate Models for Genetic Programming in Dynamic Flexible Job Shop Scheduling.
Dynamic flexible job shop scheduling (JSS) has received widespread attention from academia and industry due to its practical application value. It requires complex routing and sequencing decisions under unpredicted dynamic events. Genetic programming (GP), as a hyperheuristic approach, has been successfully applied to evolve scheduling heuristics for JSS due to its flexible representation. However, the simulation-based evaluation is computationally expensive since there are many calculations based on individuals for making decisions in the simulation. To improve training efficiency, this article proposes a novel multifidelity-based surrogate-assisted GP. Specifically, multifidelity-based surrogate models are first designed by simplifying the problem expected to be solved. In addition, this article proposes an effective collaboration mechanism with knowledge transfer for utilizing the advantages of multifidelity-based surrogate models to solve the desired problems. This article examines the proposed algorithm in six different scenarios. The results show that the proposed algorithm can dramatically reduce the computational cost of GP without sacrificing the performance in all scenarios. With the same training time, the proposed algorithm can achieve significantly better performance than its counterparts in most scenarios while no worse in others.


Development of a micro-potentiometric sensor for the microchip analysis of alkali ions.
This paper reports on the development of a micro-potentiometric sensor based on external microelectrodes introduced into a microchip. We miniaturized reference and ion-selective electrodes (ISEs) and embedded them into a plastic (PDMS) microchip; the miniaturization of ISE was attained by using a monolithic capillary-based membrane. This sensor was applied to the detection of alkali ions (Na+, K+ and NH4+) in a microflow on the microg/L level.


The influence of salicylate on platelets and whole blood adenine nucleotides.
1. The addition of sodium salicylate to freshly withdrawn human blood or native platelet-rich plasma significantly delays platelet aggregation in vitro.2. The administration of acetylsalicylic acid to human subjects also significantly delays platelet aggregation in their whole blood and there is a short delay in the formation of fibrin but this is not statistically significant.3. Salicylate, whether added to human blood in vitro as sodium salicylate or given by mouth as acetylsalicylic acid, significantly reduces the platelet clumping activity of adenosine diphosphate (ADP) added to whole blood in vitro.4. The administration of aspirin in high doses for several days produces a marked increase in the total adenine nucleotide content of whole blood. The percentage of adenosine triphosphate (ATP) was increased, that of ADP decreased while there is an obvious increase in the ATP: ADP ratio.5. There is little correlation between the plasma salicylate level and the delay aspirin produces in platelet aggregation in vitro or the changes that occur in the levels of ADP or ATP in whole blood during administration of aspirin.6. Significant correlations do occur, however, between the delay in platelet aggregation in vitro and (i) the percentage increase in the ATP concentration, (ii) the percentage decrease in ADP concentration, (iii) the percentage change in the ATP: ADP ratio observed during aspirin administration.


Awareness and utilization of peer support programs in Singapore public general hospitals.
To address the effects of acute, chronic and cumulative stress in the healthcare environment in Singapore, the Ministry of Health provided funding to develop a comprehensive crisis response management system (peer support programs/PSPs) that increases mental health awareness, provides emotional support to affected staff during work-related critical incidents and assists hospital management to better understand the emotional needs of the employees. This paper reports the awareness and utilization of PSPs in Singapore public general hospitals about one year after they were set up.


Cross-reactivity and masqueraders in seafood reactions.
Confounding variables play a significant role in many adverse seafood reactions and a clear understanding of these factors is important in properly characterizing reactions associated with potential masqueraders and mimics. Although the medical literature is replete with reviews of seafood hypersensitivity and reports of cross-reactive and newly characterized allergens, there has not been a recent effort to provide an updated overview of the several processes that may lead clinicians to draw incorrect conclusions in evaluating reported reactions to seafood. Ranging from seafood intoxications to other nonallergic or complex seafood reactions, these events can easily be misconstrued as representing a seafood IgE-mediated allergy. Among these are the more familiar topics of cross-reactivity and scombroid intoxication, and those with a still evolving understanding such as ciguatera fish poisoning and Anisakis reactions. This article seeks to provide an accessible but comprehensive summary of the relevant information surrounding these confounders in assessing adverse reactions to seafood. Such knowledge may be instrumental in unraveling complex or otherwise unclear presentations and aid clinicians in accurately evaluating and managing patients with reported seafood reactions.


Female sex and oral contraceptive use accelerate nicotine metabolism.
Several studies have reported that female smokers have a higher risk of lung cancer than male smokers. This could be related to sex differences in nicotine metabolism and related smoking behavior. This study tested the hypothesis that women metabolize nicotine more rapidly than men and that, among women, oral contraceptive users metabolize nicotine more rapidly than nonusers of oral contraceptives. Two hundred seventy-eight healthy volunteers who were twins and 16 who were siblings of twins, recruited from the Northern California Twin Registry, received an infusion of deuterium-labeled nicotine and cotinine with frequent blood sampling. The plasma clearances of nicotine and cotinine, the clearance of nicotine to cotinine (an index of cytochrome P450 [CYP] 2A6 activity), and the ratio of trans-3'-hydroxycotinine to cotinine (another indicator of CYP2A6 activity) were measured. The clearances of nicotine and cotinine, the clearance of nicotine to cotinine, and the trans-3'-hydroxycotinine/cotinine ratio were significantly higher in women than in men (nicotine clearance, 15.6 +/- 4.3 mL.min(-1).kg(-1) in men versus 18.8 +/- 6.6 mL.min(-1).kg(-1) in women; P < .001); they were also higher among women taking oral contraceptives than in those who were not taking oral contraceptives (nicotine clearance, 22.5 +/- 6.6 mL.min(-1).kg(-1) in women taking oral contraceptives versus 17.6 +/- 6.1 mL.min(-1).kg(-1) in those who were not; P < .05). Women who were menopausal or postmenopausal were not different from men. Among oral contraceptive users, nicotine metabolism was accelerated among those taking combined and estrogen-only contraceptives but not progesterone-only contraceptives. Sex hormones influence nicotine metabolism. Nicotine and cotinine metabolism is faster in women than in men and is faster in women taking oral contraceptives compared with those who are not. Accelerated nicotine metabolism appears to be a result of estrogen. Sex-related differences in nicotine clearance could affect smoking behaviors, as well as response to nicotine medications, and could be a marker for altered metabolism of nicotine-derived carcinogens.


Rat aortic smooth muscle cells become pericytes during angiogenesis in vitro.
We previously reported that the intimal endothelium of the rat aorta switches to a microvascular phenotype during angiogenesis in vitro. The microvessels formed by the rat aortic endothelium are coated with pericytes. The purpose of this study was to evaluate the relation of the pericytes to the angiogenic process and to identify the site of origin of these cells in the aortic wall. Rings of rat aorta were cultured in collagen gel under serum-free conditions. The formation of a pericyte coating around aorta-derived microvessels was evaluated by counting pericytes and microvessels in the living cultures. Pericytes and endothelial cells were studied by immunohistochemistry, lectin labeling, electron microscopy, 3H-thymidine labeling followed by autoradiography, and time-lapse video microscopy. The capacity of aortic smooth muscle cells to differentiate into pericytes was studied by coculturing intimal- or medial-derived smooth muscle cells with endothelial cells in a collagen gel overlay assay that induced reorganization of endothelial cells into microvessels. Microvessels during the early stages of angiogenesis were composed primarily of endothelial cells. As vascular proliferation decreased, the microvessels became coated with pericytes. The pericytes migrated from the root to the tip of the microvessels using the endothelium as a surface for attachment, proliferation, and contact guidance. The pericytes were continuous with the myointimal endothelial cells of the cultured aorta. Pericytes and myointimal cells were positive for alpha-smooth muscle actin and vimentin and were actively engaged in DNA synthesis. Treatment of the cultures with heparin caused a marked reduction in the number of pericytes. Smooth muscle cells isolated from the intimal aspect of the rat aorta migrated toward the endothelium and differentiated into pericytes when cocultured with microvessels formed by isolated endothelial cells in a collagen gel overlay assay. Conversely, smooth muscle cells isolated from the deep layers of the media had no significant endothelial tropism and failed to differentiate into pericytes. This study demonstrates that the rat aorta contains a subpopulation of intimal/subintimal smooth muscle cells that differentiate into pericytes during angiogenesis in vitro. These cells have a distinct endothelial tropism and respond to endothelial cues by contributing to the differentiation and maturation of microvessels. Smooth muscle cells of rat aortic intimal/subintimal origin can be used as a source of pericytes for the in vitro assembly of histotypic microvessels.


Voiding dysfunction in women with systemic lupus erythematosus.
We sought to explore bladder dysfunction in a cohort of women with systemic lupus erythematosus (SLE). We conducted a prospective study of 152 female patients with SLE during a 15-month period. The clinical status of SLE was determined according to the SLE Disease Activity Index (SLEDAI), and bladder function was evaluated by lower urinary tract symptoms and urodynamic studies. We adapted the American Urological Association (AUA) index questionnaire to assess lower urinary tract symptoms in patients, which were compared with those in 227 age-matched healthy women. The proportion of individuals reporting urinary frequency, urgency, weak urinary stream, and incomplete emptying, as well as severe lower urinary tract symptoms (AUA index score >/=20), was significantly higher in the SLE group when compared with the control group. The AUA index score showed a modest correlation with the SLEDAI score (r = 0.35, P < 0.001) but not with patient age or disease duration. There was a significant relationship between central nervous system involvement and the AUA index score. The most common urodynamic finding was a small cystometric bladder capacity (<150 ml; n = 7 patients), followed by a subnormal urinary flow rate (<12 ml/second; n = 6 patients). In 3 of 7 patients with small cystometric bladder capacities, imaging studies documented a contracted bladder with marked hydroureteronephrosis. Patients with SLE experience an increased prevalence of voiding dysfunction compared with healthy individuals. Voiding dysfunction can be attributable to either direct bladder involvement or other disease-related factors.


Heat conduction in nanoscale materials: a statistical-mechanics derivation of the local heat flux.
We derive a coarse-grained model for heat conduction in nanoscale mechanical systems. Starting with an all-atom description, this approach yields a reduced model, in the form of conservation laws of momentum and energy. The model closure is accomplished by introducing a quasilocal thermodynamic equilibrium, followed by a linear response approximation. Of particular interest is the constitutive relation for the heat flux, which is expressed nonlocally in terms of the spatial and temporal variation of the temperature. Nanowires made of copper and silicon are presented as examples.


[Diagnostic validity of palpation in horses with back problems].
There is a regularly high proportion of x-ray findings indicating a pathological deterioration of the equine vertebral column which do not correlate with the clinical symptoms. Therefore, palpation remains one of the most important clinical examination methods to determine whether or not a horse is suffering from back pain. The aim of this study was to check the validity of palpation results of horses with an assumed back problem. The palpation results of 167 horses were evaluated in retrospect: If the palpation proceeds under following conditions, positive palpation results agree with an primary back problem in about 100% of cases: 1. Hind limb lameness, being the most likely reason for misleading positive palpation results, has to be ruled out. 2. Only reproducible painful behavior is allowed to be interpreted as positive. Under these conditions a negative palpation result is correct in about 72% of cases. For the examiner it is important to keep in mind that an apparent Kissing Spine-Syndrome can exist without being palpable. By means of clinical examination it is not always possible to distinguish between painful and normal reactions. In doubtful cases diagnosis can be made by a "diagnostic therapy", or better by scintigraphy.


The added value of semimicroelectrode recording in deep brain stimulation of the subthalamic nucleus for Parkinson disease.
Accurate placement of the leads is crucial in deep brain stimulation (DBS). To optimize the surgical positioning of the lead, a combination of anatomical targeting on MRI, electrophysiological mapping, and clinical testing is applied during the procedure. Electrophysiological mapping is usually done with microelectrode recording (MER), but the relatively undocumented semimicroelectrode recording (SMER) is a competing alternative. In this study the added value and safety of SMER for optimal lead insertion in the subthalamic nucleus (STN) in a consecutive cohort of patients with Parkinson disease (PD) was assessed. Between 2001 and 2010, a consecutive single-center cohort of 46 patients with PD underwent DBS of the STN (85 lead insertions). After exclusion of 11 lead insertions for mostly technical reasons, 74 insertions were included for the assessment. Anatomical target localization was based on either 1.5-T MRI or fused 3-T MRI with CT, with reference to anterior commissure-posterior commissure coordinates. Electrophysiological mapping was performed with SMER. Intraoperative clinical testing was dominant in determining the final lead position. The target error was defined as the absolute distance between the anatomical or electrophysiological target and the final lead position. The effect of SMER on anatomical target error reduction and final target selection was analyzed. Also, the anatomical and electrophysiological target error was judged against the different imaging strategies. For safety evaluation, the adverse events related to all lead insertions were assessed. The use of SMER significantly reduced the anatomical target error from 1.7 (SD 1.6) mm to 0.8 (SD 1.3) mm (p < 0.0001). In particular, the anatomical target error based on 1.5-T MRI was significantly reduced by SMER, from 2.3 (SD 1.5) mm to 0.1 (SD 0.5) mm (p < 0.001). Anatomical target error reduction based on 3-T MRI fused with CT was not significantly influenced by SMER (p = 0.2), because the 3-T MRI-CT combination already significantly reduced the anatomical target error from 2.3 (SD 1.5) mm to 1.5 (SD 1.5) mm compared with 1.5-T MRI (p = 0.03). No symptomatic intracerebral hemorrhage was reported. Intracerebral infection was encountered in 1 patient following lead insertion. Semimicroelectrode recording has added value in targeting the STN in DBS for patients with PD based on 1.5-T MRI. The use of SMER does not significantly reduce the anatomical target error in procedures with fused 3-T MRI-CT studies and therefore might be omitted. With the absence of hemorrhagic complications, SMER-guided lead implantation should be considered a safe alternative to MER.


Contribution of 222Rn in domestic water supplies to 222Rn in indoor air in Colorado homes.
The contribution of 222Rn from domestic water wells to indoor air was investigated in a study of 28 houses near Conifer, CO. Air concentrations determined by alpha-track detectors (ATDs) and continuous radon monitors were compared with the predictions of a single-cell model. In many of the houses, the water supply was shown to contribute significantly to levels of indoor 222Rn. The data from the ATD study were augmented with a continuous monitoring study of a house near Lyons, CO. The well water in that house has the highest known concentration of 222Rn in water yet reported (93 MBq m-3). The temporal pattern in the indoor 222Rn concentration corresponds to water-use records. In general, it is difficult to quantify the proportion of indoor radon attributable to water use. Several lines of evidence suggest that the single-cell model underestimates this proportion. Continuous-monitoring data, although useful, are impractical due to the cost of the equipment. We propose a protocol for 222Rn measurement based on three simultaneous integrating radon detectors that may help estimate the proportion of indoor 222Rn derived from the water supply.


Effects of storage time and exogenous protease inhibitors on plasma protein levels.
Plasma biomarker analysis requires intact unbiased starting material. We analyzed the effects on plasma protein profiles of protease inhibitor cocktails and preprocessing storage. Plasma from 12 healthy subjects collected with and without protease inhibitors was prepared immediately and after 2 hours of room temperature storage. The samples were analyzed by a multiplexed enzyme-linked immunosorbent assay that captured 99 chemokines and cytokines. Unsupervised hierarchical analysis clustered the samples into 4 groups; one composed predominantly of samples processed immediately, another of samples processed after 2 hours, and the remaining two were a mix of immediate and 2-hour samples. The mixed and immediate group protein profiles were similar. However, among the immediate and 2-hour samples, the levels of 37 factors differed significantly: all were greater after 2 hours. The dramatic changes in protein levels during storage were independent of protease inhibitors and are likely due to cytokine production and/or release by leukocytes and platelets.