Characteristics of a new fully programmable blood sampling device for monitoring blood radioactivity during PET.
The first performance tests of a new fully programmable blood sampling device for monitoring blood radioactivity during positron emission tomography (PET) are described. Blood is withdrawn through 1-mm internal diameter tubing using an infusion pump which can be operated at rates varying from 0 to 600 ml/h. Activity in blood is measured by a 6-cm-thick bismuth germanate crystal connected to a photomultiplier tube and multichannel analyser (MCA) which are positioned within 6 cm lead shielding. Positioning of the tubing is an exact and simple procedure. The minimal readout time of the MCA is 1 s. Two independent energy windows can be set. Operation of the pump and MCA is fully controlled by a PC, i.e. sampling time, interval time and pump rate can be varied at any time during the PET scan by user-defined scripts. A number of characteristics of the new system were studied, such as sensitivity, dead time, linearity, effect of background radiation and pump rate as a function of input pressure. In addition, dispersion was measured as a function of pump rate. Finally, first clinical results were compared with manual samples. The sensitivity equalled 0.7 and 0.2 cps/Bq for 511- and 1022-keV 30% energy windows, respectively, and the system dead time was 500 ns. The system remained linear within 2% with activity concentrations up to 2.5 MBq/cc. Short-term reproducibility was better than 3% for a 1-h period. Long-term reproducibility was about 5% (ISD), which was mainly caused by variation in the diameter of the tubing. If the device was positioned in such a way that maximum shielding was directed towards the patient, the effects of background radiation from the patient on the measured activity concentration for clinically relevant conditions was minimal (<3%). Pump rate varied with input pressure, but remained constant for a given pressure. Dispersion constants smaller than 0.14 s(-1) were observed for pump rates higher than 300 ml/h, indicating that the system dispersion is small. Clinical data showed an excellent agreement to within 3% (ISD) between the results obtained with the new system and manual samples. With the continuous blood sampler radioactivity in blood can be measured accurately during the entire course of the PET scan. Furthermore, the system is fully programmable allowing adjustment of all parameters during a single PET scan.


[The specificity of the hypotensive effect of calcium antagonists in hypertension].
The purpose of the study was to define prognostic criteria that determine individual sensitivity to the hypotensive effect of slow calcium channel blockers. After two weeks of placebo therapy 45 patients suffering from essential hypertension underwent an acute pharmacological test (APT) with nifedipine in a dose of 30 mg. Then the patients received monotherapy with nitrendipine in the increasing doses for 12 weeks. The drug effect on the renin-angiotensin-aldosterone system and the level of ionized calcium in the blood serum was estimated. In 17 patients who responded to the APT by a decrease of the mean arterial pressure (MAP) by not less than 20%, arterial pressure was corrected with nitrendipine administered in a dose not exceeding 20 mg. 28 patients whose MAP dropped by 10 to 20% in response to the APT required higher drug doses (40 mg). 7 patients in whom the MAP dropped less than 10% were practically refractory to the monotherapy with nitrendipine. A close negative correlation was found between the degree of the MAP lowering in response to the APT during nitrendipine monotherapy and plasma renin activity as well as the concentration of ionized calcium in the blood serum. The low content (less than 1.0 mmol/l) of ionized calcium in the blood serum of patients suffering from essential hypertension appeared the most valuable predictor for the hypotensive effect of the slow calcium channel blockers.


Extent of surgical resection of malignant astrocytomas of the spinal cord: outcome analysis of 35 patients.
The optimal management of malignant intramedullary spinal cord astrocytomas remains controversial. Although radiotherapy has become the standard of care, the relationship between extent of resection and survival remains unclear. We report the outcomes of the surgical management of 35 malignant spinal cord astrocytomas and assess the association of extent of resection with survival after aggressive resection of these tumors. An institutional intramedullary spinal cord tumor database (1990-2002) was reviewed to identify all patients treated for malignant astrocytomas of the spinal cord (anaplastic astrocytoma [AA] or glioblastoma multiforme [GBM]). Length of survival from surgery was charted by Kaplan-Meier plots, and association of extent of resection with survival was assessed via log rank analysis for stratified covariates and Cox proportional-hazards model for continuous covariates. Twenty-seven (77%) and eight (23%) patients underwent resection of AA and GBM, respectively. Mean age was 29 +/-16 years (range, 2-61 yr). Tumor involved six +/- four vertebral levels. For AA cases, radical resection (no residual postoperative magnetic resonance enhancement) was achieved in 12 (44%) patients and subtotal resection (residual postoperative magnetic resonance enhancement) was achieved in 15 (56%). No GBM patients underwent radical resection (mean estimated resection, 70%). After surgery, two (6%) patients improved neurologically by modified McCormick score, 19 (54%) remained stable, and 14 (40%) declined. Median overall survival for AA patients was 72 months (85% at 1 yr; 59% at 5 yr). Median overall survival for GBM patients was 9 months (31% at 1 yr; 0% at 5 yr). Subtotal versus radical resection of AA was associated with decreased overall survival (38 versus 78% at 4 yr, P = 0.028). Postoperative tumor dissemination was associated with decreased survival (P = 0.004). When adjusting for multiple comparisons (P < 0.006 needed for significance), a trend of increased survival was observed with radical resection (P = 0.023). Neurological function can be preserved with aggressive resection of malignant intramedullary spinal astrocytomas; however, motor decline may be observed in many cases. Radical resection of AA was associated with a trend of increased overall survival in nondisseminated AA cases. Radical surgery and radiotherapy of GBM was associated with poor survival, similar to historical controls of diagnostic biopsy and radiotherapy. A markedly shorter survival may be expected in cases in which AA disseminates along the neuraxis. Biopsy alone may not provide the best outcomes for patients with malignant spinal cord tumors.


The British Services Dhaulagiri Medical Research Expedition 2016: a unique military and civilian research collaboration.
High-altitude environments lead to a significant physiological challenge and disease processes which can be life threatening; operational effectiveness at high altitude can be severely compromised. The UK military research is investigating ways of mitigating the physiological effects of high altitude. The British Service Dhaulagiri Research Expedition took place from March to May 2016, and the military personnel were invited to consent to a variety of study protocols investigating adaptation to high altitudes and diagnosis of high-altitude illness. The studies took place in remote and austere environments at altitudes of up to 7500 m. This paper gives an overview of the individual research protocols investigated, the execution of the expedition and the challenges involved. 129 servicemen and women were involved at altitudes of up to 7500 m; 8 research protocols were investigated. The outputs from these studies will help to individualise the acclimatisation process and inform strategies for pre-acclimatisation should troops ever need to deploy at high altitude at short notice.


Cloning and expression of cytochrome P450 enzymes catalyzing the conversion of tyrosine to p-hydroxyphenylacetaldoxime in the biosynthesis of cyanogenic glucosides in Triglochin maritima.
Two cDNA clones encoding cytochrome P450 enzymes belonging to the CYP79 family have been isolated from Triglochin maritima. The two proteins show 94% sequence identity and have been designated CYP79E1 and CYP79E2. Heterologous expression of the native and the truncated forms of the two clones in Escherichia coli demonstrated that both encode multifunctional N-hydroxylases catalyzing the conversion of tyrosine to p-hydroxyphenylacetaldoxime in the biosynthesis of the two cyanogenic glucosides taxiphyllin and triglochinin in T. maritima. This renders CYP79E functionally identical to CYP79A1 from Sorghum bicolor, and unambiguously demonstrates that cyanogenic glucoside biosynthesis in T. maritima and S. bicolor is catalyzed by analogous enzyme systems with p-hydroxyphenylacetaldoxime as a free intermediate. This is in contrast to earlier reports stipulating p-hydroxyphenylacetonitrile as the only free intermediate in T. maritima. L-3,4-Dihydroxyphenyl[3-(14)C]Ala (DOPA) was not metabolized by CYP79E1, indicating that hydroxylation of the phenol ring at the meta position, as required for triglochinin formation, takes place at a later stage. In S. bicolor, CYP71E1 catalyzes the subsequent conversion of p-hydroxyphenylacetaldoxime to p-hydroxymandelonitrile. When CYP79E1 from T. maritima was reconstituted with CYP71E1 and NADPH-cytochrome P450 oxidoreductase from S. bicolor, efficient conversion of tyrosine to p-hydroxymandelonitrile was observed.


Increased sensitivity of photosynthesis to antimycin A induced by inactivation of the chloroplast ndhB gene. Evidence for a participation of the NADH-dehydrogenase complex to cyclic electron flow around photosystem I.
Tobacco (Nicotiana tabacum var Petit Havana) ndhB-inactivated mutants (ndhB-) obtained by plastid transformation (E.M. Horvath, S.O. Peter, T. Joët, D. Rumeau, L. Cournac, G.V. Horvath, T.A. Kavanagh, C. Schäfer, G. Peltier, P. MedgyesyHorvath [2000] Plant Physiol 123: 1337-1350) were used to study the role of the NADH-dehydrogenase complex (NDH) during photosynthesis and particularly the involvement of this complex in cyclic electron flow around photosystem I (PSI). Photosynthetic activity was determined on leaf discs by measuring CO2 exchange and chlorophyll fluorescence quenchings during a dark-to-light transition. In the absence of treatment, both non-photochemical and photochemical fluorescence quenchings were similar in ndhB- and wild type (WT). When leaf discs were treated with 5 microM antimycin A, an inhibitor of cyclic electron flow around PSI, both quenchings were strongly affected. At steady state, maximum photosynthetic electron transport activity was inhibited by 20% in WT and by 50% in ndhB-. Under non-photorespiratory conditions (2% O2, 2,500 microL x L(-1) CO2), antimycin A had no effect on photosynthetic activity of WT, whereas a 30% inhibition was observed both on quantum yield of photosynthesis assayed by chlorophyll fluorescence and on CO2 assimilation in ndhB-. The effect of antimycin A on ndhB- could not be mimicked by myxothiazol, an inhibitor of the mitochondrial cytochrome bc1 complex, therefore showing that it is not related to an inhibition of the mitochondrial electron transport chain but rather to an inhibition of cyclic electron flow around PSI. We conclude to the existence of two different pathways of cyclic electron flow operating around PSI in higher plant chloroplasts. One of these pathways, sensitive to antimycin A, probably involves ferredoxin plastoquinone reductase, whereas the other involves the NDH complex. The absence of visible phenotype in ndhB- plants under normal conditions is explained by the complement of these two pathways in the supply of extra-ATP for photosynthesis.


Improved accuracy of acetabular component position and leg lengths with the two-incision total hip replacement technique.
We sought to determine whether a non-originating surgeon early in his experience with the MIS two-incision technique for THA could place components with appropriate accuracy. Thirty five consecutive hip replacements performed between July of 2003 and March of 2004 using the two-incision technique were closely matched by height and weight with an equal number of control small incision arthroplasties. The two-incision surgeries were performed after the surgeons twentieth case and prior to the surgeons fifty sixth two-incision case. We assessed immediate postoperative films for leg length discrepancy and acetabular position, and compared the two groups by chi squared analysis. Both leg length accuracy and abduction angle were significantly superior with the two-incision technique. No arthroplasty performed with the two-incision technique had a leg length discrepancy of more than 10 mm, versus five in the control group (p<0.025). The abduction of one cup fell outside the 35-49 range in the two-incision group, while ten did so in the control group (p<0.01). With this study, we have shown that a non-originating surgeon early in his experience with the MIS two-incision technique can place acetabular components and equalise leg lengths with superior precision. We believe the fluoroscopic control and supine position greatly enhance the surgeons accuracy, and more than compensate for the potentially decreased direct vision with this technique.


Diffusion of butyrate through pig colonic mucus in vitro.
Using a modified equilibrium dialysis cell the rate of diffusion of butyrate through pig colonic mucus has been compared with that through other gels and unstirred layers. Relative diffusion coefficients were calculated for each layer. Layers of 8% polyacrylamide, and of caecal, mid-colonic and terminal colonic mucus, had coefficients that were 50-60% of the apparent free diffusion coefficient for butyrate, determined using layers made up of Millipore filters alone. The apparent free diffusion coefficients for butyrate (layers of agarose or filters) were 70% of previously determined values in the literature. This discrepancy can be explained by elements of the experimental procedure. All mucus layers differed significantly from layers of 2% agarose and Millipore filters but were not significantly different from layers of 8% polyacrylamide or from each other. Diffusion coefficients for butyrate in the mucus samples correlated with water content and carbohydrate content but had no relationship to protein content. The rate of diffusion of butyrate in colonic mucus layers was significantly reduced when compared with unstirred layers (P less than 0.05). Whether this has an effect on the butyrate supply to colonocytes in vitro and whether mucus in colonic disease behaves differently are subjects for further investigation.


Cloning and sequence determination of a complementary DNA related to human liver microsomal cytochrome P-450 S-mephenytoin 4-hydroxylase.
A cDNA sequence related to the human cytochrome P-450 responsible for S-mephenytoin 4-hydroxylation (P-450MP) has been isolated from a human liver bacteriophage lambda gt11 library with antibodies specific for P-450MP. The total length of the cDNA is 2.5 kilobases (kb), of which there is a 1.6-kb EcoRI fragment coding for all but five amino acids corresponding to the N-terminus of the protein and including a small noncoding region at the 3' end. This 1.6-kb fragment has been sequenced and used as a probe to analyze human genomic DNA and liver RNA. The sequence shows extensive sequence similarity with that of rabbit liver cytochrome P-450 progesterone 21-hydroxylase [Tukey, R. H., Okino, S., Barnes, H., Griffin, K. J., & Johnson, E. F. (1985) J. Biol. Chem. 260, 13347-13354], and this cDNA, like the rabbit clone, appears to be part of a multigene family. At least two liver mRNA species, 2.2 kb and 3.5 kb, hybridize to the cDNA sequence. The cloning of this gene should aid in analyzing the molecular basis for the genetic polymorphism of S-mephenytoin 4-hydroxylation reported in humans.


How do we ensure that trainees learn to perform biliary sphincterotomy safely, appropriately, and effectively?
Sphincterotomy is a high-risk procedure with considerable complications. Trainees should learn and understand the basics of endoscopic retrograde cholangiopancreatography and sphincterotomy to ensure good clinical outcomes. Teaching of sphincterotomy usually involves supervised hands-on clinical practice with patients. Proper positioning of the endoscope allows for correct orientation with the papilla, and performing the cut along the "ideal" biliary axis optimizes results and reduces complications. Learning and practicing sphincterotomy can be supplemented by simulator models. The Neo-Papilla model uses a modified chicken heart attached to the porcine ex vivo model and allows for cutting of actual tissue. The mechanical simulator allows trainees to practice cutting an artificial papilla marked with the "perfect" axis to understand the proper sphincterotomy technique. Understanding the indications and contraindications helps with appropriate application of sphincterotomy. Objective criteria should be available for assessing performance. Improved technique and avoiding a deviated cut may improve overall results and prevent complications.


[Effects of Preoperative Immune and Nutritional Indices on the Prognosis of pStage Ⅱ and Ⅲ Colorectal Cancer].
We examined the relationship between immune and nutritional indices and the outcome of colorectal cancer patients at our hospital. We retrospectively analyzed 196 patients with pStage Ⅱand Ⅲcolorectal cancer who underwent curative surgery in our institution between 2007 and 2013. The evaluation items were immune and nutritional indices, such as neutrophil/lym- phocyte ratio(NLR), platelet/lymphocyte ratio(PLR), lymphocyte/monocyte ratio(LMR), and the Onodera nutritional index (PNI). Moreover, we analyzed the relationship between immune and nutritional indices and outcome, overall survival(OS), and recurrence-free survival(RFS). Univariate and multivariate analyses showed that low LMR was significantly related to high mortality. Univariate analysis showed that high NLR, low LMR, and low PNIwere significantly related to a decrease in OS. Multivariate analysis showed that PNIwas an independent predictor of OS. LMR is suggested as a new predictor for postoperative complications. PNIis suggested as a new prognostic factor in pStageⅡand Ⅲcolorectal cancer.


[Multi-drug efflux pumps and antibiotic resistance in Acinetobacter baumannii].
Bacterial multi-drugs systems contribute to the development of multi-resistance patterns of Acinetobacter baumannii, a nosocomial pathogen of increasing importance due to its emerging resistance to carbapenems. The multi-resistance phenomena is generated by a combination of mechanisms, one of which the efflux pump system. Many of these multiresistant isolates of A. baumannii harbor genes for the AdeABC multi-drug efflux system, related with resistance to various groups of antibacterial agents, including tygecicline and meropenem. Inhibition of these systems would allow to increase the efficacy of this antimicrobial. This review focuses on the multi-drug efflux pump system of A. baumannii with special emphasis in the AdeABC system.


Divergent roles of Salmonella pathogenicity island 2 and metabolic traits during interaction of S. enterica serovar typhimurium with host cells.
The molecular mechanisms of virulence of the gastrointestinal pathogen Salmonella enterica are commonly studied using cell culture models of infection. In this work, we performed a direct comparison of the interaction of S. enterica serovar Typhimurium (S. Typhimurium) with the non-polarized epithelial cell line HeLa, the polarized cell lines CaCo2, T84 and MDCK, and macrophage-like RAW264.7 cells. The ability of S. Typhimurium wild-type and previously characterized auxotrophic mutant strains to enter host cells, survive and proliferate within mammalian cells and deploy the Salmonella Pathogenicity Island 2-encoded type III secretion system (SPI2-T3SS) was quantified. We found that the entry of S. Typhimurium into polarized cells was much more efficient than entry into non-polarized cells or phagocytic uptake. While SPI2-T3SS dependent intracellular proliferation was observed in HeLa and RAW cells, the intracellular replication in polarized cells was highly restricted and not affected by defective SPI2-T3SS. The contribution of aromatic amino acid metabolism and purine biosynthesis to intracellular proliferation was distinct in the various cell lines investigated. These observations indicate that the virulence phenotypes of S. Typhimurium are significantly affected by the cell culture model applied.


[Genetic diversity of major capsid genes (g23) of T4-type bacteriophages in natural environments--a review].
During the past two decades, the genetic diversities of bacterial and fungal communities and their relationship with the inhabited environments were intensively studied with the development of molecular biological techniques. However, although bacteriophages are ubiquitous and more abundant than their hosts in biosphere, their diversity is still little known. In this paper, we targeted the major capsid genes (g23) of T4-type bacteriophages and reviewed the recent progress on their genetic diversity. The distribution of g23 of T4-type bacteriophages was distinctly different among natural environments of marines, lakes and paddy fields, and the majority of g23 were grouped into several novel clusters according to their obtained environments. In addition, several research tips and future research tendencies for the study of environmental g23 were also addressed.


Assessment of the flow velocity of blood cells in a microfluidic device using joint spectral and time domain optical coherence tomography.
Although Doppler optical coherence tomography techniques have enabled the imaging of blood flow in mid-sized vessels in biological tissues, the generation of velocity maps of capillary networks remains a challenge. To better understand the origin and information content of the Doppler signal from small vessels and limitations of such measurements, we used joint spectral and time domain optical coherence tomography to monitor the flow in a model, semitransparent microchannel device. The results obtained for Intralipid, whole blood, as well as separated red blood cells indicate that the technique is suitable to record velocity profiles in vitro, in a range of microchannel configurations.


The influenza pandemic of 1918-1919 in two remote island nations: Iceland and New Zealand.
Nations varied in their experience of, and response to, the 1918-19 influenza pandemic. Island communities can provide unique opportunities to study the epidemiology of infectious diseases. We aimed to compare the epidemiology and public health response to this pandemic in two remote island nations, on opposite sides of the globe: Iceland and New Zealand (NZ). Historical accounts in both nations were reviewed, along with recent analysis of the pandemics impact and course. Marked similarities were noted in epidemic timing, failure of border control, shape of epidemic curves, and delayed use of public health interventions. However, amongst the exposed European populations, Iceland experienced a significantly higher mortality rate (830 vs 550 per 100,000) compared to NZ (rate ratio: 1.5, 95%CI: 1.4-1.6). There is evidence that some public health measures in specific areas of both nations resulted in lower mortality rates. In particular, Iceland's use of travel restrictions and ship quarantining, appeared to protect 36% of the population. The epidemiology of the 1918-19 influenza pandemic was fairly similar for the exposed European populations of Iceland and NZ. Nevertheless, major differences were the significantly higher overall mortality rate in Iceland and the success of Iceland's use of travel restrictions.


Disruption of the vacuolar-type H+-ATPase complex in liver causes MTORC1-independent accumulation of autophagic vacuoles and lysosomes.
The vacuolar-type H+-translocating ATPase (v-H+-ATPase) has been implicated in the amino acid-dependent activation of the mechanistic target of rapamycin complex 1 (MTORC1), an important regulator of macroautophagy. To reveal the mechanistic links between the v-H+-ATPase and MTORC1, we destablilized v-H+-ATPase complexes in mouse liver cells by induced deletion of the essential chaperone ATP6AP2. ATP6AP2-mutants are characterized by massive accumulation of endocytic and autophagic vacuoles in hepatocytes. This cellular phenotype was not caused by a block in endocytic maturation or an impaired acidification. However, the degradation of LC3-II in the knockout hepatocytes appeared to be reduced. When v-H+-ATPase levels were decreased, we observed lysosome association of MTOR and normal signaling of MTORC1 despite an increase in autophagic marker proteins. To better understand why MTORC1 can be active when v-H+-ATPase is depleted, the activation of MTORC1 was analyzed in ATP6AP2-deficient fibroblasts. In these cells, very little amino acid-elicited activation of MTORC1 was observed. In contrast, insulin did induce MTORC1 activation, which still required intracellular amino acid stores. These results suggest that in vivo the regulation of macroautophagy depends not only on v-H+-ATPase-mediated regulation of MTORC1.


Cytokine profiling of tumor interstitial fluid of the breast and its relationship with lymphocyte infiltration and clinicopathological characteristics.
The tumor microenvironment is composed of many immune cell subpopulations and is an important factor in the malignant progression of neoplasms, particularly breast cancer (BC). However, the cytokine networks that coordinate various regulatory events within the BC interstitium remain largely uncharacterized. Moreover, the data obtained regarding the origin of cytokine secretions, the levels of secretion associated with tumor development, and the possible clinical relevance of cytokines remain controversial. Therefore, we profiled 27 cytokines in 78 breast tumor interstitial fluid (TIF) samples, 43 normal interstitial fluid (NIF) samples, and 25 matched serum samples obtained from BC patients with Luminex xMAP multiplex technology. Eleven cytokines exhibited significantly higher levels in the TIF samples compared with the NIF samples: interleukin (IL)-7, IL-10, fibroblast growth factor-2, IL-13, interferon (IFN)γ-inducible protein (IP-10), IL-1 receptor antagonist (IL-1RA), platelet-derived growth factor (PDGF)-β, IL-1β, chemokine ligand 5 (RANTES), vascular endothelial growth factor, and IL-12. An immunohistochemical analysis further demonstrated that IL-1RA, IP-10, IL-10, PDGF-β, RANTES, and VEGF are widely expressed by both cancer cells and tumor-infiltrating lymphocytes (TILs), whereas IP-10 and RANTES were preferentially abundant in triple-negative breast cancers (TNBCs) compared to Luminal A subtype cancers. The latter observation corresponds with the high level of TILs in the TNBC samples. IL-1β, IL-7, IL-10, and PDGFβ also exhibited a correlation between the TIF samples and matched sera. In a survival analysis, high levels of IL-5, a hallmark TH2 cytokine, in the TIF samples were associated with a worse prognosis. These findings have important implications for BC immunotherapy research.


Epigenetic profiling of human brain differential DNA methylation networks in schizophrenia.
Epigenetics of schizophrenia provides important information on how the environmental factors affect the genetic architecture of the disease. DNA methylation plays a pivotal role in etiology for schizophrenia. Previous studies have focused mostly on the discovery of schizophrenia-associated SNPs or genetic variants. As postmortem brain samples became available, more and more recent studies surveyed transcriptomics of the diseases. In this study, we constructed protein-protein interaction (PPI) network using the disease associated SNP (or genetic variants), differentially expressed disease genes and differentially methylated disease genes (or promoters). By combining the different datasets and topological analyses of the PPI network, we established a more comprehensive understanding of the development and genetics of this devastating mental illness. We analyzed the previously published DNA methylation profiles of prefrontal cortex from 335 healthy controls and 191 schizophrenic patients. These datasets revealed 2014 CpGs identified as GWAS risk loci with the differential methylation profile in schizophrenia, and 1689 schizophrenic differential methylated genes (SDMGs) identified with predominant hypomethylation. These SDMGs, combined with the PPIs of these genes, were constructed into the schizophrenic differential methylation network (SDMN). On the SDMN, there are 10 hypermethylated SDMGs, including GNA13, CAPNS1, GABPB2, GIT2, LEFTY1, NDUFA10, MIOS, MPHOSPH6, PRDM14 and RFWD2. The hypermethylation to differential expression network (HyDEN) were constructed to determine how the hypermethylated promoters regulate gene expression. The enrichment analyses of biochemical pathways in HyDEN, including TNF alpha, PDGFR-beta signaling, TGF beta Receptor, VEGFR1 and VEGFR2 signaling, regulation of telomerase, hepatocyte growth factor receptor signaling, ErbB1 downstream signaling and mTOR signaling pathway, suggested that the malfunctioning of these pathways contribute to the symptoms of schizophrenia. The epigenetic profiles of DNA differential methylation from schizophrenic brain samples were investigated to understand the regulatory roles of SDMGs. The SDMGs interplays with SCZCGs in a coordinated fashion in the disease mechanism of schizophrenia. The protein complexes and pathways involved in SDMN may be responsible for the etiology and potential treatment targets. The SDMG promoters are predominantly hypomethylated. Increasing methylation on these promoters is proposed as a novel therapeutic approach for schizophrenia.


High hydrostatic pressure upon the vasa vasorum of the greater saphenous and splenic vein walls: a comparative study.
Hypoxia and high hydrostatic pressure can induce an increase in the thickness of the tunica media and intima; secondary vasa vasorum (VV) increase to fit the remodeling of the vessel wall. We aimed to investigate the impact of high hydrostatic pressure on VV in the varicose greater saphenous veins (VGSVs) and diseased splenic veins (DSVs). We collected 34 VGSVs and DSVs. Thirty-four normal greater saphenous veins (GSVs) and splenic veins (SVs) were also collected (control group). Samples were cut into slices, and observed under both light and electron microscopy. The mean density and cross-sectional areas of the VV in the adventitia were measured. In both VGSVs and DSVs, VV density increased, in the adventitia and exterior tunica media, offering an intensive linear distribution. However, sporadic distribution of the interior tunica media and intima were seen on light microscopy. The integrated structure of the cell nucleus of endothelial cells in VV, normal morphology and distribution of chromatin, partially hyperchromatic mitochondria matrix, fuzzy or fractured mitochondria cristae, and medullary cristae changes were observed by electron microscopy. Mean density and cross-sectional areas of VV in the adventitia of GSVs and SVs were significantly different. Under high hydrostatic pressure conditions, the number of VV were increased in the wall of VGSVs and DSVs. There was heterogeneity between both types of veins. The splenic vein has a higher number of VV, but the greater saphenous vein has a higher average cross-sectional area. The same ultrastructural changes are seen in the endothelial cells of the VV in both vessels.


Comparative chromosome painting of pronghorn (Antilocapra americana) and saola (Pseudoryx nghetinhensis) karyotypes with human and dromedary camel probes.
Pronghorn (Antilocapridae, 2n = 58) and saola (Bovidae, 2n = 50) are members of Pecora, a highly diversified group of even-toed hoofed mammals. Karyotypes of these species were not involved in chromosome painting studies despite their intriguing phylogenetic positions in Pecora. To trace the chromosome evolution during very fast radiation of main families from the common Pecoran ancestor, high-resolution comparative chromosome maps of pronghorn and saola with human (HSA) and dromedary camel (CDR) painting probes were established. The human and dromedary camel painting probes revealed 50 and 64 conserved segments respectively in the pronghorn genome, while 51 and 63 conserved segments respectively in the saola genome. Integrative analysis with published comparative maps showed that inversions in chromosomes homologous to CDR19/35/19 (HSA 10/20/10), CDR12/34/12 (HSA12/22/12/22), CDR10/33/10 (HSA 11) are present in representatives of all five living Pecoran families. The pronghorn karyotype could have formed from a putative 2n = 58 Pecoran ancestral karyotype by one fission and one fusion and that the saola karyotype differs from the presumed 2n = 60 bovid ancestral karyotype (2n = 60) by five fusions. The establishment of high-resolution comparative maps for pronghorn and saola has shed some new insights into the putative ancestral karyotype, chromosomal evolution and phylogenic relationships in Pecora. No cytogenetic signature rearrangements were found that could unite the Antilocapridae with Giraffidae or with any other Pecoran families. Our data on the saola support a separate position of Pseudorigyna subtribe rather than its affinity to either Bovina or Bubalina, but the saola phylogenetic position within Bovidae remains unresolved.


Management of advanced breast cancer with endocrine therapy: the role of the primary healthcare team.
Of the 25,000 new cases of breast cancer annually in the UK, at least half will develop metastases at some time. Approximately one-third of these will be hormone-sensitive and thus suitable for endocrine manipulation. Such patients may live for several years. Care should be patient-centred and evidence-based, with primary care playing an integral part in the multidisciplinary team. Quality of life is paramount, and largely determines decisions to change endocrine therapy. Tamoxifen remains the first choice treatment, but newer agents, in particular the oral aromatase inhibitors, have successfully extended the range of therapy available. Ovarian ablation can be effectively achieved using LHRH analogues, which have the advantage of being reversible if ineffective. In future, further improvements in outcome are likely with new pure antioestrogens and the possibility of sequential or combined use of existing drugs.


Inhibition of Stearoyl-CoA Desaturase 1 expression in human lung adenocarcinoma cells impairs tumorigenesis.
Saturated (SFA) and monounsaturated (MUFA) fatty acids, the most abundant fatty acid species, have many divergent biological effects including the regulation of cell proliferation, programmed cell death and lipid-mediated cytotoxicity. Their distribution is regulated by Stearoyl-CoA Desaturases (SCD), the enzymes that convert SFA into MUFA. A positive correlation between high levels of tissue MUFA and several types of cancer has been reported, but a causal relationship between the function of SCD1, the main human SCD isoform, and cancer development has not yet been firmly established. Here we report that the stable knockdown of SCD1 gene expression in A549 human lung adenocarcinoma cells decreased the ratio MUFA/SFA in total lipids and inhibited the incorporation of glucose into cell lipids. Cell proliferation and anchorage-independent growth were considerably decreased in SCD1-depleted cells, whereas the rate of apoptosis was elevated, with respect to control A549 cells. In addition, phosphorylation of Akt-Ser473 and GSK-3beta-Ser9 was found notably impaired in SCD1-ablated A549 cells. Interestingly, the effects of SCD1 blockade on Akt activation, cancer cell growth and apoptosis could not be reversed by exogenously added oleic acid. Remarkably, the reduction of SCD1 expression in lung cancer cells significantly delayed the formation of tumors and reduced the growth rate of tumor xenografts in mice. Our study demonstrates that SCD1 activity regulates Akt activation and determines the rate of cell proliferation, survival and invasiveness in A549 cancer cells and shows, for the first time, that SCD1 is a key factor in the regulation of tumorigenesis in vivo.


Identification of Maximal Running Intensities During Elite Hurling Match-Play.
Young, D, Malone, S, Beato, M, Mourot, L, and Coratella, G. Iidentification of maximal running intensities during elite hurling match-play. J Strength Cond Res 34(9): 2608-2617, 2020-The current study aimed to describe the duration-specific running intensities of elite hurling players during competition with respect to position using a rolling average method. Global positioning systems (10-Hz Viper; STATSport, Viper, Newry, Northern Ireland) were used to collect data from 36 elite hurling players across 2 seasons. Players were categorized according to playing positions (full-backs, half-backs, midfielders, half-forwards, and full-forwards). A total of 230 full match samples were obtained from 22 competitive games for analysis. The velocity-time curve was analyzed using a rolling average method, in which the maximum relative total distance (TD; m·min), high-speed running distance (HSR; m·min), and sprint distance (SD; m·min) intensities were calculated across 10 different rolling time durations (1-10 minutes) within each game. There were large to very large (effect sizes [ES] = 0.66-4.33) differences between 1 minute rolling averages and all other durations for TD, HSR, and SD. However, pairwise comparisons between 6 and 10 minutes for TD, HSR, and SD were smaller and more variable (ES = 0.07, trivial to ES = 0.85, moderate). Half-backs, midfielders, and half-forwards achieved a higher maximal relative TD and HSR in all duration-specific fields when compared with full-backs and full-forwards. No positional difference was observed in 1- and 2-minute durations for SD. Because the rolling average duration increased the maximum TD, HSR and SD running intensities decreased across all positions. These data provide knowledge of the peak running intensities of elite hurling competition and can be used to design training activities to sufficiently prepare players for these "worst-case scenarios."


Catalytic activity of human indoleamine 2,3-dioxygenase (hIDO1) at low oxygen.
A cytokine-inducible extrahepatic human indoleamine 2,3-dioxygenase (hIDO1) catalyzes the first step of the kynurenine pathway. Immunosuppressive activity of hIDO1 in tumor cells weakens host T-cell immunity, contributing to the progression of cancer. Here we report on enzyme kinetics and catalytic mechanism of hIDO1, studied at varied levels of dioxygen (O2) and L-tryptophan (L-Trp). Using a cytochrome b5-based activating system, we measured the initial rates of O2 decay with a Clark-type oxygen electrode at physiologically-relevant levels of both substrates. Kinetics was also studied in the presence of two substrate analogs: 1-methyl-L-tryptophan and norharmane. Quantitative analysis supports a steady-state rather than a rapid equilibrium kinetic mechanism, where the rates of individual pathways, leading to a ternary complex, are significantly different, and the overall rate of catalysis depends on contributions of both routes. One path, where O2 binds to ferrous hIDO1 first, is faster than the second route, which starts with the binding of L-Trp. However, L-Trp complexation with free ferrous hIDO1 is more rapid than that of O2. As the level of L-Trp increases, the slower route becomes a significant contributor to the overall rate, resulting in observed substrate inhibition.


Comparisons of morphologic characteristics between thin-flap LASIK and SBK.
To compare the morphologic characteristics between thin-flap laser in situ keratomileusis (LASIK) and Sub-Bowman keratomileusis (SBK), and to evaluate the uniformity of flap and to explore the correlative factors of corneal flap thickness. A prospective, randomized, comparative clinical study was performed in Department of Ophthalmology, West Hospital of China, Chengdu, Sichuan Province, China. Totally 59 patients 114 eyes underwent LASIK or SBK to correct myopia, 29 patients 57 eyes underwent SBK, 30 patients 57 eyes underwent LASIK. Anterior optical coherence tomography(OCT) was used to measure corneal flap thickness in all the patients 1 week after surgeries, 16 positions were set to be measured in each eye. Comparisons of flap thickness in each group and between 2 groups were evaluated. Correlative factors of flap thickness were evaluated. Coefficient of variation (CV, s/×%) in SBK group were lesser than that in LASIK group, t test showed there was significant statistical difference between 2 groups(P=0.000). Comparisons of the difference of 2 paired positions(temporal-nasal; superior-inferior) showed there were no significant differences in each group, but between 2 groups, there were statistical significance of value of difference (D-value) of superior and inferior positions between SBK and LASIK group(P=0.036). Linear regression analysis of correlative factors of flap thickness showed there were no statistic significances related to central corneal thickness(CCT)(P=0.060, t=1.921) and corneal curvature(P=0.083, t=1.766). SBK is better than LASIK in creating much uniform corneal flap. There was no evidence showing correlations between flap thickness and CCT or corneal curvature.


Psychostimulant-induced behavioral sensitization depends on nicotinic receptor activation.
Animal studies have shown that nicotine and psychostimulant drugs (amphetamine and cocaine) share the property of inducing long-lasting behavioral and neurochemical sensitization, which is thought to contribute to their addictive properties. Neuroplasticity subserving learning and memory mechanisms is considered to be involved in psychostimulant-induced sensitization and addiction behavior. Because nicotinic receptors in the brain play a role in the storage of drug-related information underlying reinforcement learning, we evaluated the possibility that activation of central nicotinic receptors may underlie psychostimulant-induced sensitization. Repeated exposure of rats to nicotine profoundly enhanced the psychomotor effects of nicotine and amphetamine 3 weeks after nicotine pretreatment. Moreover, the nicotinic receptor antagonist mecamylamine completely blocked the induction, but not the long-term expression, of behavioral sensitization to amphetamine in amphetamine-pretreated rats. Mecamylamine also prevented the development of cocaine-induced behavioral sensitization. Behavioral sensitization induced by nicotine, amphetamine, or cocaine was associated with an increase in the electrically evoked release of [(3)H]dopamine from nucleus accumbens slices. Coadministration of mecamylamine during pretreatment with nicotine, amphetamine, or cocaine prevented the development of this long-term hyperreactivity of nucleus accumbens dopamine neurons. Similarly, the high-affinity non-alpha7 subtype nicotinic receptor antagonist dihydro-beta-erythroidine prevented the development of amphetamine-induced behavioral and neurochemical sensitization. These data indicate that nicotinic receptor activation (by endogenously released acetylcholine) is a common denominator initiating neuroplasticity involved in the development of amphetamine, as well as cocaine-induced sensitization.


Rendering the world unsafe: 'vulnerability' as western discourse.
Disasters seem destined to be major issues of academic enquiry in the new century if for no other reason than that they are inseparably linked to questions of environmental conservation, resource depletion and migration patterns in an increasingly globalised world. Unfortunately, inadequate attention has been directed at considering the historical roots of the discursive framework within which hazard is generally presented, and how that might reflect particular cultural values to do with the way in which certain regions or zones of the world are usually imagined. This paper argues that tropicality, development and vulnerability form part of one and the same essentialising and generalising cultural discourse that denigrates large regions of world as disease-ridden, poverty-stricken and disaster-prone.


Colovesical and rectovesical fistulas.
Colovesical or rectovesical fistulas develop because of a congenital abnormality, a traumatic incident, or an underlying disease, such as inflammation or cancer. Inflammation, either diverticulitis or Crohn's disease, was the cause of fistula in 73 (67%) of 109 patients with colovesical or rectovesical fistulas seen at the Mayo Clinic, Rochester, Minn, between 1965 and 1980. Sixty-six of the 73 patients had colovesical fistulas, and 64% of them underwent resection and anastomosis in one stage. Cancer of the cervix, prostate, bladder, colon, or rectum was responsible for 27 fistulas, equally divided between colovesical and rectovesical types. Only seven (26%) could be treated by one-stage anterior resection. Fifteen patients required fecal diversion with or without resection of bowel. At follow-up, 80 patients were alive; only two (one with prostatic cancer and one with cervical cancer) had recurrence of fistula.


Early virologic response and IL28B polymorphisms in patients with chronic hepatitis C genotype 3 treated with peginterferon alfa-2a and ribavirin.
Polymorphisms of the IL28B gene (rs12979860 and rs8099917) are associated with high sustained virological response (SVR) rates in HCV genotype 1 patients. This study analyzes the impact of these IL28B polymorphisms on early treatment response (weeks 2 and 4) and SVR in HCV genotype 3 patients. rs12979860 and rs8099917 were analyzed by the Step-OnePlus Real-time PCR system in 71 out of 72 Caucasian HCV genotype 3 patients participating, at our center, in a randomized study comparing 400mg with 800 mg ribavirin/day. HCV RNA was determined at weeks 2 and 4 of 180 μg/week peginterferon alfa-2a/ribavirin treatment. Sixty-nine patients completed the treatment and follow-up. rs12979860 genotyping revealed that 27 (37.5%) patients had C/C, 39 (54.2%) T/C, and 5 (6.9%) T/T. Thirteen patients (18.1%) became HCV RNA negative at week 2 and an additional 30 (41.7%) at week 4 (rapid virologic response; RVR); thus a total of 43 had a RVR (C/C: 77.8%; T/C or T/T: 50.0%). Irrespective of the ribavirin dose, the viral load decline was larger than in those with the T allele (T/C or T/T) (week 2: 4.46; [0.36-6.02] median; [range] vs. 3.50; [0.14-5.62]; log IU HCV-RNA/ml; p<0.001; week 4: 4.97; [1.21-6.20] vs. 4.49; [1.16-6.23]; p=0.003). Despite the faster initial viral response in C/C carriers, SVR rates were not different compared to T-allele carriers. Results of the SNP in the rs8099917 region were similar. IL28B polymorphisms modulate early virologic response to peginterferon/ribavirin treatment. In contrast to HCV genotype 1 patients, no effect on SVR rates was observed in genotype 3 patients. The clinical relevance of an earlier viral decline in C/C patients needs to be determined.


Propagation of Helmholtz-Gauss beams in absorbing and gain media.
The propagation of Helmholtz-Gauss beams in media exhibiting loss or gain is studied. The general expressions for the field propagation, the time-averaged power on propagation, the trajectory of the beam centroid, the beam spreading, the nondiffracting distance, and the far field are derived and discussed. Explicit expressions of these parameters for Bessel-Gauss and cosine-Gauss beams are included. The general expressions can be applied straightforwardly to describe the propagation of Mathieu-Gauss and parabolic-Gauss beams in complex media as well.


Assessing statistical similarity in dietary intakes of women of reproductive age in Bangladesh.
Women of reproductive age (WRA) need adequate nutrient intakes to sustain a healthy pregnancy, support fetal growth, and breastfeed after childbirth. However, data on women's dietary intake in low- and middle-income countries (LMICs) are limited, and assessment of differences between dietary intakes of pregnant or lactating women compared with that of nonpregnant, nonlactating (NPNL) women is untested. Using single, multiple-pass 24-h dietary recall data from a sample of WRA residing in rural Bangladesh, we examined women's dietary intakes for energy, protein, calcium, iron, vitamin A, and dietary diversity for three groups: NPNL (n = 2,903), pregnant (n = 197), and lactating women (n = 944). We used equivalence testing to examine similarity in adjusted intakes for pregnant versus NPNL women and lactating versus NPNL women with a predetermined equivalence threshold based on recommendations specific for each reproductive stage. On average, both pregnant and lactating women had insufficient intakes for all dietary measures. Although statistically significant differences were observed between pregnant and NPNL women for energy intake and dietary diversity and between lactating and NPNL women for energy and protein intake, the magnitudes of these differences were too small to reject equivalence. Statistical similarity was also evident in all micronutrients and dietary diversity for both two-group comparisons. Understanding statistical differences and similarities between dietary measures of women in distinct reproductive stages has important implications for the relevance, appropriateness, and evaluation of maternal diet-enhancing interventions in LMICs, especially during pregnancy and lactation, when demand for macronutrients and micronutrients is elevated.


Molecular identification of burn spot disease syndrome agent Fusarium avenaceum in Turkish crayfish Pontastacus leptodactylus populations.
In this study, we aimed to isolate and identify the causative agent of burn spot disease syndrome from crayfish populations within Turkish water bodies. Crayfish Pontastacus leptodactylus samples were captured from Bayramşah Reservoir, Karaidemir Reservoir and Keban Reservoir (Turkey). Fungi samples were taken from symptomatic crayfish and cultured in potato dextrose agar, and DNAs were extracted. Species-specific PCR primers (FAF1 and FAR) were used in order to amplify the internal transcribed spacer (ITS) region of fungal DNA. Sequences obtained from the amplified regions were analyzed using GenBank and identified. As a result, a molecular diagnosis of the fungi samples was made, and the pathogen was identified as Fusarium avenaceum. In addition, a challenge test with healthy crayfish was conducted to evaluate the pathogenicity of the fungi. F. avenaceum was reisolated from experimentally infested individuals, and thusly we confirmed the disease. It was concluded that F. avenaceum causes burn spot disease syndrome in P. leptodactylus.


Vitamin D status and its relationship with parathyroid hormone and bone mineral status in older adolescents.
Osteoporosis is an important contributor to the global burden of disease, and in the UK alone results in one in three women and one in twelve men aged >50 years experiencing a fragility fracture. Optimising peak bone mass in early adulthood is thought to reduce osteoporosis risk by offsetting bone losses in later life. Ensuring sufficient vitamin D status (measured as 25-hydroxyvitamin D (25OHD) in plasma), among other factors, is believed to facilitate the achievement of optimum peak bone mass. Lower 25OHD is associated with a higher plasma concentration of parathyroid hormone (PTH). As PTH is associated with increased bone turnover and bone loss, maintenance of sufficient 25OHD is thought to have a protective effect on bone health. However, there is a lack of consensus internationally on what constitutes an optimum 25OHD concentration, and values between 30 and 80 nmol/l have been suggested. These values have been based on findings from various studies in adults in which PTH has been observed to plateau at a 25OHD concentration of >30 nmol/l; however, not all studies have found such a plateau. Although studies in younger adolescents (14-16 years) have shown an inverse relationship between PTH and 25OHD, the concentration of 25OHD required for achievement of optimum peak bone mass is unknown. The present review examines the evidence defining vitamin D insufficiency thresholds, and the relevance of such thresholds to adolescent bone health.


The structure of the Autism Diagnostic Interview-Revised: diagnostic and phenotypic implications.
Multivariate statistics can assist in refining the nosology and diagnosis of pervasive developmental disorders (PDD) and also contribute important information for genetic studies. The Autism Diagnostic Interview-Revised (ADI-R) is one of the most widely used assessment instruments in the field of PDD. The current study investigated its factor structure and convergence with measures of adaptive, language, and intellectual functioning. Analyses were conducted on 1,861 individuals with PDD between the ages of 4 and 18 years (mean = 8.3, SD = 3.2). ADI-R scores were submitted to confirmatory factor analysis (CFA) and exploratory factor analysis (EFA). Analyses were conducted according to verbal status (n = 1,329 verbal, n = 532 nonverbal) and separately for algorithm items only and for all items. ADI-R scores were correlated with scores on measures of adaptive, language, and intellectual functioning. Several factor solutions were examined and compared. CFAs suggested that two- and three-factor solutions were similar, and slightly superior to a one-factor solution. EFAs and measures of internal consistency provided some support for a two-factor solution consisting of social and communication behaviors and restricted and repetitive behaviors. Measures of functioning were not associated with ADI-R domain scores in nonverbal children, but negatively correlated in verbal children. Overall, data suggested that autism symptomatology can be explained statistically with a two-domain model. It also pointed to different symptoms susceptible to be helpful in linkage analyses. Implications of a two-factor model are discussed.


Recurrent postural vasovagal syncope: sympathetic nervous system phenotypes.
The pathophysiology of vasovagal syncope is poorly understood, and the treatment usually ineffective. Our clinical experience is that patients with vasovagal syncope fall into 2 groups, based on their supine systolic blood pressure, which is either normal (>100 mm Hg) or low (70-100 mm Hg). We investigated neural circulatory control in these 2 phenotypes. Sympathetic nervous testing was at 3 levels: electric, measuring sympathetic nerve firing (microneurography); neurochemical, quantifying norepinephrine spillover to plasma; and cellular, with Western blot analysis of sympathetic nerve proteins. Testing was done during head-up tilt (HUT), simulating the gravitational stress of standing, in 18 healthy control subjects and 36 patients with vasovagal syncope, 15 with the low blood pressure phenotype and 21 with normal blood pressure. Microneurography and norepinephrine spillover increased significantly during HUT in healthy subjects. The microneurography response during HUT was normal in normal blood pressure and accentuated in low blood pressure phenotype (P=0.05). Norepinephrine spillover response was paradoxically subnormal during HUT in both patient groups (P=0.001), who thus exhibited disjunction between nerve firing and neurotransmitter release; this lowered norepinephrine availability, impairing the neural circulatory response. Subnormal norepinephrine spillover in low blood pressure phenotype was linked to low tyrosine hydroxylase (43.7% normal, P=0.001), rate-limiting in norepinephrine synthesis, and in normal blood pressure to increased levels of the norepinephrine transporter (135% normal, P=0.019), augmenting transmitter reuptake. Patients with recurrent vasovagal syncope, when phenotyped into 2 clinical groups based on their supine blood pressure, show unique sympathetic nervous system abnormalities. It is predicted that future therapy targeting the specific mechanisms identified in the present report should translate into more effective treatment.


Sense and nonsense about the Fenn effect.
One of the most influential papers in muscle physiology was published by W. O. Fenn (J. Physiol. London 58: 175-203) in 1923. Fenn determined the quantitative relationship between muscle energy liberation and work performance. Despite the importance of this work, the implications of Fenn's observations are sometimes misunderstood. In this review article, Fenn's experiments are reexamined and the Fenn effect delineated in light of the viscoelastic model of muscle contraction prevalent in the 1920's. The generality of Fenn's results and conclusions are considered in view of more recent results from skeletal and cardiac muscle. Factors responsible for the considerable deviation from Fenn's original results are discussed. Much of the confusion surrounding the generality of Fenn's observations seems to be attributable to the problem of determining an energetic base line for comparing isotonic and isometric contractions. Mommaerts (Physiol. Rev. 49: 427-508, 1969) has suggested the use of an equivalent force base line, which appears to be useful and unifying concept.


Vitreo-retinal traction and anastrozole use.
This study tested a prediction stemming from the hypothesis that anastrozole users experience heightened vitreo-retinal traction. This hypothesis was based on the knowledge that menopause increases the risk of intraocular tractional events such as posterior vitreous detachments (PVDs). Retinal thickness was measured for 3 groups of amenorrheic women: (1) anastrozole users and (2) tamoxifen users undergoing adjuvant therapy for early-stage breast cancer, and (3) control subjects not using hormonal medication. Foveal shape indices were derived for subjects without PVDs. For anastrozole users, the distance to the temporal side of the fovea became less than the distance to the nasal side at a sufficient height above the foveal base. This effect did not exist for control subjects; the between-group difference was appreciable. Results concerning tamoxifen users were inconclusive. The foveas of women using anastrozole appear to be subjected to more tractional force than are the foveas of women not using any hormonal medication.


Low-order aberration correction of the TMT tertiary mirror prototype based on a warping harness.
A warping harness is proposed to simply and efficiently correct low-order aberrations that occur during manufacturing and operation of a telescope. For the Thirty Meter Telescope (TMT) tertiary mirror, the issue to be solved by the warping harness is particularly challenging due to its complicated load conditions and limited mounting space. In this study, first, a new type of whiffletree-based warping harness configuration applied to a ¼-prototype TMT tertiary mirror is presented and optimized using finite element analysis (FEA) to improve the output precision of the moment actuator. Next, based on the new configuration, a simulation method for a correction process is proposed. The results show that the root mean square value of the mirror-surface error converged from 64.9 to 25.4 nm after correction, which satisfied the requirement document of TMT. Finally, combined with the analysis and calculation results, the moment actuator testing system with high-precision displacement-force-strain is established to assess the system errors. The tests of the moment actuator displacement, stress, strain-output precision, linearity, and repeatability are completed, and all errors were found to be within a controllable range. The results show the validity and feasibility of the designed warping harness, which can prove its applicability in more complicated conditions and, to a certain degree, broaden the application scope of the warping harness.


Recent trends in hospital admissions and outcomes of cardiac Chagas disease in the United States.
Chagas disease (CD), caused by Trypanosoma cruzi, has been increasingly encountered as a cause of cardiovascular disease in the United States. We aimed to examine trends of hospital admissions and cardiovascular outcomes of cardiac CD (CCD). Search of 2003-2011 Nationwide Inpatient Sample database identified 949 (age 57±16 years, 51% male, 72.5% Hispanic) admissions for CCD. A significant increase in the number of admissions for CCD was noted during the study period (OR=1.054; 95% CI=1.028-1.081; P< 0.0001); 72% were admitted to Southern and Western hospitals. Comorbidities included hypertension (40%), coronary artery disease (28%), hyperlipidemia (26%), tobacco use (12%), diabetes (9%), heart failure (5%) and obesity (2.2%). Cardiac abnormalities noted during hospitalization included atrial fibrillation (27%), ventricular tachycardia (23%), sinoatrial node dysfunction (5%), complete heart block (4%), valvular heart disease (6%)] and left ventricular aneurysms (5%). In-hospital mortality was 3.2%. Other major adverse events included cardiogenic shock in 54 (5.7%), cardiac arrest in 30 (3.2%), acute heart failure in 88 (9.3%), use of mechanical circulatory support in 29 (3.1%), and acute stroke in 34 (3.5%). Overall, 63% suffered at least one adverse event. Temporary (2%) and permanent (3.5%) pacemakers, implantable cardioverter defibrillators (10%), and cardiac transplant (2.1%) were needed for in-hospital management. Despite the remaining concerns about lack of awareness of CCD in the US, an increasing number of hospital admissions were reported from 2003-2011. Serious cardiovascular abnormalities were highly prevalent in these patients and were frequently associated with fatal and nonfatal complications.


Production of tumor necrosis factor during murine listeriosis.
During a lethal murine infection with the gram-positive bacterium, Listeria monocytogenes, a factor appears in the serum that is capable of lysing certain tumor cells in vitro. The levels of this serum cytotoxic factor increase with the progression of morbidity. Neutralization of Listeria-induced cytotoxic factor activity with a monospecific antiserum to recombinant murine tumor necrosis factor (TNF) revealed that the cytotoxic factor was antigenically indistinguishable from natural TNF present in endotoxin-induced tumor necrosis serum. In contrast to a lethal infection, no TNF activity was detected in sera of mice throughout a sublethal infection. However, shortly after initiation of a sublethal infection, mice acquire a greatly enhanced capacity to produce serum TNF in response to an intravenous injection of endotoxin. Cultures of unfractionated spleen cells from mice with ongoing Listeria infections produced TNF when incubated in the absence, but not in the presence, of antibiotics. However, such antibiotic-treated cultures could be stimulated with endotoxin to produce substantially higher TNF yields than spleen cells of uninfected mice. It is also shown that intravenous infusion of anti-recombinant murine TNF IgG into sublethally infected mice results in increased Listeria proliferation in spleens and livers, and ultimately in death from an overwhelming infection.


MRSA Prevalence in Preoperative S. aureus Nasal Culture Isolates is Significantly Different From a Traditional Hospital-wide Antibiogram.
Hospital antibiograms, because they are typically derived from samples obtained from hospitalized patients, may overestimate the prevalence of methicillin resistance in S. aureus in individuals presenting to the hospital for surgery. Because hospital antibiograms are commonly used to justify empiric perioperative prophylactic antibiotic selection prior to surgery, this may lead to unnecessary treatment with broad-spectrum antibiotics such as vancomycin. In a single-institution study, we observed that in our hospital antibiogram the proportion of S. aureus that are methicillin-resistant (MRSA) was significantly higher (45%) than isolates in preoperative nasal cultures obtained at the same hospital in outpatients prior to their lower extremity joint replacement surgery (13%): mean difference 0.32, [95% CI 0.25, 0.39], p <0.0001. These data suggest that hospital antibiograms may overstate the true prevalence of MRSA in those at risk for MRSA surgical site infections who present from the outpatient setting.


Sgo1 establishes the centromeric cohesion protection mechanism in G2 before subsequent Bub1-dependent recruitment in mitosis.
Bub1 was one of the first protein kinases identified as a component of the spindle-assembly checkpoint, a surveillance mechanism that delays anaphase onset until all chromosomes are stably attached to spindle microtubules. Whereas the kinase activity of Bub1 is not required for checkpoint function in yeast, its requirement in mammalian cells is still unclear. Using a complementation assay with bona fide BUB1-null mouse embryonic fibroblasts, we show that the kinase activity of Bub1 is not required for checkpoint function or chromosome alignment. Its activity is, however, required for centromeric localisation of Sgo1, a known protector of centromeric cohesion. Despite the absence of Sgo1 from mitotic centromeres in cells devoid of Bub1 activity, centromeric cohesion is still maintained until anaphase. An explanation for this comes from observations showing that Sgo1 is first recruited to centromeric heterochromatin in G2, but then becomes diffusely localised throughout the nucleus in early prophase, before returning to centromeres later in prophase. Importantly, whereas centromeric localisation of Sgo1 in prophase is dependent on the kinase activity of Bub1, its recruitment to centromeric heterochromatin in G2 is not. Rather, the localisation of Sgo1 in G2 is abolished when heterochromatin protein 1 is not bound to centromeric heterochromatin. Thus, it seems that Sgo1 sets up the centromeric protection mechanism in G2, but that its Bub1-dependent localisation to centromeres during mitosis is not required to maintain cohesion.


Expression of Phosphorylated KIT in Canine Mast Cell Tumor.
Canine cutaneous mast cell tumor (MCT) is the most common canine skin tumor and exhibits variable biologic behavior. Signaling through the KIT receptor tyrosine kinase promotes cellular proliferation and survival and has been shown to play a role in MCT progression. Despite investigations into numerous biomarkers and the proposal of several grading schemas, no single marker or grading system can accurately predict outcome in canine MCT. The first aim of this study was to develop an immunohistochemical assay to measure phosphorylated KIT (pKIT) to investigate its association with 2 commonly used grading systems and other established prognostic markers for canine MCT. Thirty-four archived MCTs were evaluated for expression of pKIT and Ki-67, KIT localization, mitotic count, mutations in exons 8 and 11 in c-kit, and grading by the Patnaik and 2-tier systems. Expression of pKIT was significantly ( P < .05) correlated with the 2-tier grading scheme and c-kit mutation. Correlation approached significance ( P = .06) with Mitotic Index (MI) and Ki-67. An additional aim was to determine whether pKIT labeling provides a pharmacodynamic marker for predicting response to the receptor tyrosine kinase inhibitor toceranib (TOC). MCTs from 4 of 7 patients demonstrated a partial response to TOC. pKIT expression was assessed by immunohistochemistry in biopsies obtained before and 6 hours after the patients were treated with TOC. Reduced pKIT expression after TOC treatment was demonstrated in 3 of the 4 patients with a partial response compared to 1 of the 3 nonresponders. Collectively, these results demonstrate that immunohistochemical detection of pKIT may be a clinically relevant assay to evaluate the activation status of the major oncogenic pathway in canine MCT.


The landscape of clinical trials in nephrology: a systematic review of Clinicaltrials.gov.
Well-designed trials are of paramount importance in improving the delivery of care to patients with kidney disease. However, it remains unknown whether contemporary clinical trials within nephrology are of sufficient quality and quantity to meet this need. Systematic review. Studies registered with ClinicalTrials.gov. Interventional (ie, nonobservational) studies (both randomized and nonrandomized) registered between October 2007 and September 2010 were included for analysis. Studies were reviewed independently by physicians and classified by clinical specialty. Nephrology versus cardiology versus other trials. Select clinical trial characteristics. Of 40,970 trials overall, 1,054 (2.6%) were classified as nephrology. Most nephrology trials were for treatment (75.4%) or prevention (15.7%), with very few diagnostic, screening, or health services research studies. Most nephrology trials were randomized (72.3%). Study designs included 24.9% with a single study group, 64.0% that included parallel groups, and 9.4% that were crossover trials. Nephrology trials, compared with 2,264 cardiology trials (5.5% overall), were more likely to be smaller (64.5% vs 48.0% enrolling≤100 patients), phases 1-2 (29.0% vs 19.7%), and unblinded (66.2% vs 53.3%; P<0.05 for all). Nephrology trials also were more likely than cardiology trials to include a drug intervention (72.4% vs 41.9%) and less likely to report having a data monitoring committee (40.3% vs 48.5%; P<0.05 for all). Finally, there were few trials funded by the National Institutes of Health (NIH; 3.3%, nephrology; 4.2%, cardiology). Does not include all trials performed worldwide, and frequent categorization of funding source as university may underestimate NIH support. Critical differences remain between clinical trials in nephrology and other specialties. Improving care for patients with kidney disease will require a concerted effort to increase the scope, quality, and quantity of clinical trials within nephrology.


Ventricular pacing with premature excitation for treatment of hypertensive-cardiac hypertrophy with cavity-obliteration.
Hypertensive left ventricular hypertrophy with supranormal systolic ejection and distal cavity obliteration (HHCO) can result in debilitating exertional fatigue and dyspnea. Dual-chamber pacing with ventricular preactivation generates discoordinate contraction, which can limit cavity obliteration and thereby increase potential ejection reserve. Accordingly, we hypothesized that pacing may improve exercise tolerance long-term in this syndrome. Dual-chamber pacemakers were implanted in 9 patients with exertional dyspnea caused by HHCO. Intrinsic atrial rate was sensed, and ventricular preactivation was achieved by shortening the atrial-ventricular delay. Pacing was on or off for successive 3-month periods (randomized, double-blind, crossover design), followed by 6 additional pacing-on months. Metabolic exercise testing, quality-of-life assessment, and rest and dobutamine-stress echocardiographic/Doppler data were obtained. After 3 months of pacing-on, exercise duration rose from 324+/-133 to 588+/-238 s (mean+/-SD; P=0.001, with 7 of 9 patients improving >/=30%), and maximal oxygen consumption increased from 13.6+/-2.9 to 16.7+/-3.3 mL of O(2). min(-1). kg(-1) (P<0.02). Both parameters were little changed from baseline during the pacing-off period. Improved exercise capacity persisted at 1-year follow-up. Clinical symptoms and activities of daily living improved during the pacing-on period and stayed improved at 1 year, but they were little changed during the pacing-off period. Despite similar basal values, stroke volume (P<0.001) and cardiac output (P<0.02) increased with dobutamine stimulation 2 to 3 times more after 1 year of follow-up as compared with baseline. Long-term dual-chamber pacing can improve exercise capacity, cardiac reserve, clinical symptoms, and activities of daily living in patients with HHCO. This therapy may provide a novel alternative for patients in whom traditional pharmacological treatment proves inadequate.


Egg marketing in national supermarkets: products, packaging, and prices--part 3.
As part of a national retail egg quality study, the variety of shell eggs and egg products offered for sale, type of packaging, and price relationships were compared in five major metropolitan regions. A total of 81 stores in 28 cities were sampled in California (CA), Illinois (IL), North Carolina (NC), Texas (TX), and New England (NE). Data were recorded for the variety of brands, sizes, white or brown shell eggs, specialty eggs, liquid or frozen eggs, carton sizes, package labeling and coding, and price relationships of shell eggs, liquid, and frozen egg products displayed for sale. The total variety of shell eggs displayed per store was the greatest for CA and NE stores. Stores in CA and TX offered more (P < 0.05) variety of white shell eggs than did stores in the other states, whereas stores in NE displayed the greatest variety (P < 0.05) of brown shell eggs. The average number of liquid and frozen egg products was highest (P < 0.05) for NC stores. Packaging type, USDA labeling, and carton coding differed somewhat among states. The price per one dozen cartons of all white shell egg sizes was highest (P < 0.05) in CA stores, and the average liquid plus frozen egg product prices were higher in CA and NE stores compared to the other states. However, the ratio of liquid and frozen product prices to all large shell egg prices was among the lowest for CA and NC stores. These data indicate that product selection, packaging, and consumer prices for shell eggs and egg products varied considerably across five separate regions of the country.


Access to school-based eye health programs: a qualitative case study, Bogotá, Colombia.
To identify barriers and enablers to accessing school-based eye health programs in Bogotá, Colombia. We undertook a qualitative case study that explored how structural factors, and social and cultural norms influence access to school-based eye health programs. We conducted focus groups discussions and interviews with a purposive sample of 37 participants: government stakeholders (n = 4), representatives from nongovernmental organizations (n = 3), and an eye-care practitioner, as well as teachers (n = 7), a school nurse, parents (n = 7), and children (n = 14) from private and public schools. Data were analyzed using a priori themes from the availability, accessibility, acceptability and quality framework. Routine vision screening in schools is not currently provided nor is there a budget to support it. Lack of collaboration between the health and education ministries and the absence of national planning affected the delivery of eye care in schools. Factors related to acceptability of school-based eye health programs included: poor acceptance of training teachers as vision screeners; stigma related to wearing spectacles; and distrust of health services. The cost of spectacles and poor access to eye health information were identified as barriers to positive child eye health outcomes by socioeconomically disadvantaged parents and children. Our findings suggest the need for a national school eye health plan and improved cooperation between health and education ministries. Interventions to improve trust in health services, tackle the lack of human resources while respecting professional qualifications, and raise awareness of the importance of eye health are recommended.


The effects of tissue expansion on skin lymph flow and lymphatics: an experimental study in rabbits.
Histomorphological, physiological, and biomechanical changes of the skin and tissue that are being expanded have been extensively studied in the past. The purpose of this study was to investigate whether the skin lymph flow and lymphatics may be influenced during tissue expansion, and also to study different pattern of vascular supply of skin on lymph flow during tissue expansion. The skin lymph flow was quantitatively assessed by 99mTc-dextran lymphoscintigraphy, and the structure of dermal lymphatics was evaluated by histological examination in 12 rabbits. Lymphoscintigraphic results showed that lymph flow is significantly reduced both in expanded and non-expanded (sham-operated) skin. Histologically, we saw widespread lymphatic distension in both expanded and non-expanded skin. There were no quantitative differences in the number of lymphatic vessels compared with control skin. We did not see any lymphatic capillaries in the subdermal capsule of either expanded or non-expanded skin. These results show that lymphostasis has an obstructive (mechanical) aetiology, and the tissue expander itself reduces the lymph flow regardless of expansion, and interferes with the formation of new lymphatic vessels.


Distinct expression pattern and post-transcriptional regulation of cell cycle genes in the glandular epithelia of avian ovarian carcinomas.
The cell cycle system is controlled in a timely manner by three groups of cyclins, cyclin dependent kinases and cyclin dependent kinase inhibitors. Abnormal alterations of cell cycle regulatory mechanisms are a common feature of many diseases including numerous tumor types such as ovarian cancer. Although a variety of cell cycle regulatory genes are well known in mammalian species including human and mice, they are not well studied in avian species, especially in laying hens which are recognized as an excellent animal model for research relevant to human ovarian carcinogenesis. Therefore, in the present study, we focused on comparative expression and regulation of expression of candidate genes which might be involved in the cell cycle program in surface epithelial ovarian cancer in laying hens. Our current results indicate that expression levels of cell cycle gene transcripts are greater in cancerous as compared to normal ovaries. In particular, cyclin A2 (CCNA2), CCND1, CCND2, CCND3, CCNE2, cyclin dependent kinase 1 (CDK1), CDK3, CDK5, cyclin dependent kinases inhibitor 1A (CDKN1A) and CDKN1B were upregulated predominantly in the glandular epithelia of cancerous ovaries from laying hens. Further, several microRNAs (miRs), specifically miR-1798, miR-1699, miR-223 and miR-1744 were discovered to influence expression of CCND1, CCNE2, CDK1, and CDK3 mRNAs, respectively, via their 3'-UTR which suggests that post-transcriptional regulation of gene expression influences their expression in laying hens. Moreover, miR-1626 influenced CDKN1A expression and miR-222, miR-1787 and miR-1812 regulated CDKN1B expression via their 3'-UTR regions. Collectively, results of the present study demonstrate increased expression of cell cycle-related genes in cancerous ovaries of laying hens and indicate that expression of these genes is post-transcriptionally regulated by specific microRNAs.


Type of Multimorbidity and Complementary and Alternative Medicine Use among Adults.
Objective. To examine the association between type of multimorbidity and CAM use among adults with multimorbidity. Methods. The current study used a cross-sectional design with retrospective data from 2012 National Health Interview Survey. Multimorbidity was classified into two groups: (1) adults with coexisting physical and mental illnesses and (2) adults with two or more chronic physical illnesses only. CAM use was measured using a set of 18 variables. Logistic regression and multinomial logistic regressions were used to assess the association between the type of multimorbidity and ever used CAM, CAM use in the past 12 months, and type of CAM. Results. Overall, 31.2% of adults with coexisting physical and mental illnesses and 20.1% of adults with only physical illnesses used CAM in the past 12 months. Adults with coexisting physical and mental illnesses were more likely to ever use CAM (AOR = 1.68, 95% CI = 1.49, 1.90), use CAM in the past 12 months (AOR = 1.32, 95% CI = 1.15, 1.52), and use mind-body therapies in the past 12 months (AOR = 1.36, 95% CI = 1.16, 1.59) compared to adults with only physical illnesses. Conclusion. Multimorbidity of chronic physical and mental illnesses was associated with higher CAM use.


Sexual function 6 months after first delivery.
To explore the association of anal sphincter laceration and sexual function 6 months postpartum in the Childbirth and Pelvic Symptoms (CAPS) cohort. The primary CAPS study, a prospective cohort study, was designed to estimate the postpartum prevalence and incidence of urinary and fecal incontinence. Three cohorts of new mothers (vaginal delivery with a third- or fourth-degree anal sphincter tear, vaginal delivery without a third- or fourth-degree anal sphincter tear, and cesarean delivery without labor) were compared at 6 months postpartum. Sexual function was assessed with the Pelvic Organ Prolapse/Urinary Incontinence/Sexual Function Short Form Questionnaire (PISQ-12). Urinary and fecal incontinence were assessed using the Medical Epidemiological and Social Aspects of Aging questionnaire and the Fecal Incontinence Severity Index, which is embedded within the Modified Manchester Health Questionnaire. Most women (459 [90%]) of those with partners reported sexual activity at the 6-month visit. Fewer women whose delivery was complicated by anal sphincter laceration reported sexual activity when compared with those who delivered vaginally without sphincter laceration (88 compared with 94%, P=.028). The mean PISQ-12 score (39+/-4) did not differ between delivery groups (P=.92). Pain (responses of "sometimes," "usually," or "always") during sex affected one of three sexually active women (164 [36%]). At 6 months postpartum, primiparous women who delivered with anal sphincter laceration are less likely to report sexual activity. II.


Mesenchymal stem cells ameliorate inflammation in an experimental model of Crohn's disease via the mesentery.
Mesenchymal stem cells (MSCs) are novel therapeutics for treatment of Crohn's disease. However, their mechanism of action is unclear, especially in disease-relevant chronic models of inflammation. Thus, we used SAMP-1/YitFc, a chronic and spontaneous murine model of small intestinal inflammation, to study the therapeutic effect and mechanism of human bone marrow-derived MSCs (hMSC). hMSC immunosuppressive potential was evaluated through in vitro mixed lymphocyte reaction, ELISA, macrophage co-culture, and RT-qPCR. Therapeutic efficacy and mechanism in SAMP were studied by stereomicroscopy, histopathology, MRI radiomics, flow cytometry, RT-qPCR, small animal imaging, and single-cell RNA sequencing (Sc-RNAseq). hMSC dose-dependently inhibited naïve T lymphocyte proliferation in MLR via PGE 2 secretion and reprogrammed macrophages to an anti-inflammatory phenotype. hMSC promoted mucosal healing and immunologic response early after administration in SAMP model of chronic small intestinal inflammation when live hMSCs are present (until day 9) and resulted in complete response characterized by mucosal, histological, immunologic, and radiological healing by day 28 when no live hMSCs are present. hMSC mediate their effect via modulation of T cells and macrophages in the mesentery and mesenteric lymph nodes (mLN). Sc-RNAseq confirmed the anti-inflammatory phenotype of macrophages and identified macrophage efferocytosis of apoptotic hMSCs as a mechanism of action that explains their long-term efficacy. hMSCs result in healing and tissue regeneration in a chronic model of small intestinal inflammation. Despite being short-lived, exert long-term effects via macrophage reprogramming to an anti-inflammatory phenotype. Single-cell RNA transcriptome datasets are deposited in an online open access repository 'Figshare' (DOI: https://doi.org/10.6084/m9.figshare.21453936.v1 ).


Summary of presentations at the NIH/NIAID New Humanized Rodent Models 2007 Workshop.
It has long been recognized that a small animal model susceptible to HIV-1 infection with a functional immune system would be extremely useful in the study of HIV/AIDS pathogenesis and for the evaluation of vaccine and therapeutic strategies to combat this disease. By early 2007, a number of reports on various rodent models capable of being infected by and responding to HIV including some with a humanized immune system were published. The New Humanized Rodent Model Workshop, organized by the Division of AIDS (DAIDS), National Institute Allergy and Infection Diseases (NIAID), NIH, was held on September 24, 2007 at Bethesda for the purpose of bringing together key model developers and potential users. This report provides a synopsis of the presentations that discusses the current status of development and use of rodent models to evaluate the pathogenesis of HIV infection and to assess the efficacy of vaccine and therapeutic strategies including microbicides to prevent and/or treat HIV infection.


Structure-based design of robust glucose biosensors using a Thermotoga maritima periplasmic glucose-binding protein.
We report the design and engineering of a robust, reagentless fluorescent glucose biosensor based on the periplasmic glucose-binding protein obtained from Thermotoga maritima (tmGBP). The gene for this protein was cloned from genomic DNA and overexpressed in Escherichia coli, the identity of its cognate sugar was confirmed, ligand binding was studied, and the structure of its glucose complex was solved to 1.7 Angstrom resolution by X-ray crystallography. TmGBP is specific for glucose and exhibits high thermostability (midpoint of thermal denaturation is 119 +/- 1 degrees C and 144 +/- 2 degrees C in the absence and presence of 1 mM glucose, respectively). A series of fluorescent conjugates was constructed by coupling single, environmentally sensitive fluorophores to unique cysteines introduced by site-specific mutagenesis at positions predicted to be responsive to ligand-induced conformational changes based on the structure. These conjugates were screened to identify engineered tmGBPs that function as reagentless fluorescent glucose biosensors. The Y13C*Cy5 conjugate is bright, gives a large response to glucose over concentration ranges appropriate for in vivo monitoring of blood glucose levels (1-30 mM), and can be immobilized in an orientation-specific manner in microtiter plates to give a reversible response to glucose. The immobilized protein retains its response after long-term storage at room temperature.


Local routing algorithms based on Potts neural networks.
A feedback neural approach to static communication routing in asymmetric networks is presented, where a mean field formulation of the Bellman-Ford method for the single unicast problem is used as a common platform for developing algorithms for multiple unicast, multicast and multiple multicast problems. The appealing locality and update philosophy of the Bellman-Ford algorithm is inherited. For all problem types the objective is to minimize a total connection cost, defined as the sum of the individual costs of the involved arcs, subject to capacity constraints. The methods are evaluated for synthetic problem instances by comparing to exact solutions for cases where these are accessible, and else with approximate results from simple heuristics. In general, the quality of the results are better than those of the heuristics. Furthermore, the computational demands are modest, even when the distributed nature of the the approach is unexploited numerically.


Impact of engineered surface microtopography on biofilm formation of Staphylococcus aureus.
The surface of an indwelling medical device can be colonized by human pathogens that can form biofilms and cause infections. In most cases, these biofilms are resistant to antimicrobial therapy and eventually necessitate removal or replacement of the device. An engineered surface microtopography based on the skin of sharks, Sharklet AF, has been designed on a poly(dimethyl siloxane) elastomer (PDMSe) to disrupt the formation of bacterial biofilms without the use of bactericidal agents. The Sharklet AF PDMSe was tested against smooth PDMSe for biofilm formation of Staphylococcus aureus over the course of 21 days. The smooth surface exhibited early-stage biofilm colonies at 7 days and mature biofilms at 14 days, while the topographical surface did not show evidence of early biofilm colonization until day 21. At 14 days, the mean value of percent area coverage of S. aureus on the smooth surface was 54% compared to 7% for the Sharklet AF surface (p<0.01). These results suggest that surface modification of indwelling medical devices and exposed sterile surfaces with the Sharklet AF engineered topography may be an effective solution in disrupting biofilm formation of S. aureus.


Structure and expression in Escherichia coli of a cloned rat interferon-alpha gene.
DNA synthesized by in vitro transcription on rat interferon (IFN) mRNA has been cloned and amplified as recombinant DNA. The nucleotide sequence of these rat IFN cDNA clones revealed i. the partial presence of the coding region of the gene and ii all cDNA clones were derived from the same subtype of rat IFN-alpha mRNA. Purified inserted fragments were used as a hybridisation probe against chromosomal "Southern blots" to show that at least twelve rat IFN-alpha-related sequences are present in the genome. A lambda-linked rat gene library was screened with the cDNA probes, resulting in an equivalent number of rat IFN-alpha-related hybrid phages. By use of a 3'-noncoding region as a probe, the chromosomal counterpart of the cDNA clones could be detected and the nucleotide sequence of its coding region has been determined. Expression of the coding region in E. coli yielded biologically active IFN, when tested for in vitro or in vivo antiviral activity.


Conserved hydrophobic amino acid residues in the N-terminal region of DnaA protein are involved in DnaA-DnaA interaction.
We previously reported that a leucine-zipper-like structure (I26, L33 and L40) located in the N-terminal region of DnaA is essential for the duplex opening at oriC by DnaA. In this study, we focused on three other conserved hydrophobic amino acid residues, L3, L10 and L17, and examined the function of DnaA proteins mutated in these amino acid residues. DnaA427 (L17S) and DnaA413 (L3S, L10S and L17S) were inactive for oriC DNA replication both in vitro and in vivo. Although these mutant DnaA proteins maintained their binding activities for both ATP and oriC, they were unable to induce the opening of duplex DNA at oriC. Glutathione-S-transferase (GST)-fused wild-type DnaA interacted with wild-type DnaA but not with DnaA427 and DnaA413. Based on these results, we propose that conserved hydrophobic amino acid residues in the N-terminal region of DnaA are involved in DnaA oligomerization, in which DnaA-DnaA interaction is required.


Herpes simplex encephalitis in glioma patients: a challenging diagnosis.
In recent years, herpes simplex encephalitis (HSE) has been reported with increasing frequency in settings of immunosuppression, such as acquired immunodeficiency, transplantation and cancer. As observed, in immunocompromised individuals HSE presents peculiar clinical and paraclinical features, and poorer prognosis. Here we describe a retrospective series of seven cases of HSE in patients with high-grade glioma (HGG), collected among three institutions in a 5-year period (during this time, a total of 1750 patients with HGG were treated). Diagnosis of the condition was particularly challenging due to the confounding clinical presentation and the atypical biological findings. As a result, antiviral treatment was started with a sharp delay compared with immunocompetent hosts. Prognosis was poor, with high short-term mortality and severe residual disability in survivors. The substantial incidence of HSE observed in our centres together with the difficulty in diagnosing the condition suggest that the incidence of this complication may be highly underestimated. The aim of our report is to strengthen the observation of HSE in patients with HGG and outline the key elements that may allow its diagnosis.


Current trends in nanobiosensor technology.
The development of tools and processes used to fabricate, measure, and image nanoscale objects has lead to a wide range of work devoted to producing sensors that interact with extremely small numbers (or an extremely small concentration) of analyte molecules. These advances are particularly exciting in the context of biosensing, where the demands for low concentration detection and high specificity are great. Nanoscale biosensors, or nanobiosensors, provide researchers with an unprecedented level of sensitivity, often to the single molecule level. The use of biomolecule-functionalized surfaces can dramatically boost the specificity of the detection system, but can also yield reproducibility problems and increased complexity. Several nanobiosensor architectures based on mechanical devices, optical resonators, functionalized nanoparticles, nanowires, nanotubes, and nanofibers have been demonstrated in the lab. As nanobiosensor technology becomes more refined and reliable, it is likely it will eventually make its way from the lab to the clinic, where future lab-on-a-chip devices incorporating an array of nanobiosensors could be used for rapid screening of a wide variety of analytes at low cost using small samples of patient material.


Methyl bromide production from dissolved organic matter under simulated sunlight irradiation and the important effect of ferric ions.
The oceans play an important role in the biogeochemical cycle of methyl bromide (CH3Br), as not only the sinks but also the sources. However, many uncertainties exist regarding the way of CH3Br generation in the marine environment. To illustrate the possibility of photochemical formation of CH3Br in saline water, its generation in bromide aqueous solution containing humic acid (HA) and ferric ions (Fe(iii)) was investigated. CH3Br was obviously generated after irradiation, and its amounts increased with increasing HA concentration from 0.82 to 12.2 mgC L-1. Fe(iii) significantly promoted CH3Br production, and the described production process in the presence of Fe(iii) was pH-dependent, decreasing with the increase of pH. Finally, the concentrations of CH3Br in natural coastal seawater and seawater with HA were measured, and the results showed that CH3Br was significantly generated under irradiation. Our results suggest that the photochemical process of dissolved organic matter may be one source of CH3Br in the marine environment.


Development of a targeted method for twenty-three metabolites related to polyphenol gut microbial metabolism in biological samples, using SPE and UHPLC-ESI-MS/MS.
An increasing number of studies have concerned the profiling of polyphenol microbial metabolites, especially in urine or plasma, but only a few have regarded their accurate quantification. This study reports on a new ultra-performance liquid chromatography tandem mass spectrometry method with electrospray ionisation (UHPLC-ESI-MS/MS) using a simple clean-up step with solid phase extraction (SPE) and validation on different biological matrices. The method was tested with spiked samples of liver, heart, kidneys, brain, blood and urine. The purification procedure, after the evaluation of three different cartridges, makes it possible to obtain cleaner samples and better quantification of putative trace metabolites, especially related to dietary studies, with concentrations below ng/g in tissue and for urine and blood, starting from ng/ml. Limits of detection and linear range were also assessed using mixed polyphenol metabolite standards. Short chromatographic separation was carried out for 23 target compounds related to the polyphenol microbial metabolism, coupled with a triple quadrupole mass spectrometer for their accurate quantification. By analysing different spiked biological samples we were able to test metabolite detection in the matrix and validate the overall recovery of the method, from purification to quantification. The method developed can be successfully applied and is suitable for high-throughput targeted metabolomics analysis related to nutritional intervention, or the study of the metabolic mechanism in response to a polyphenol-rich diet.


The action of trimebutine maleate on gastrointestinal motility is mediated by opiate receptors in human subjects.
The effects of trimebutine on interdigestive motor activity of the small intestine and the colon were studied in healthy volunteers. Trimebutine induced in all subjects phase III-like motor complexes, which were similar to spontaneously occurring phase III complexes of the interdigestive cycle in antrum and duodenum. Trimebutine-induced complexes were suppressed by naloxone. Trimebutine and/or naloxone had no consistent effect on interdigestive contractile activity in the colon.


Major biocontrol of plant tumors targets tRNA synthetase.
Crops can be devastated by pathogenic strains of Agrobacterium tumefaciens that cause crown gall tumors. This devastation can be prevented by the nonpathogenic biocontrol agent A. radiobacter K84, which prevents disease by production of the "Trojan horse" toxin agrocin 84, which is specifically imported into tumorgenic A. tumefaciens strains to cause cell death. We demonstrate that this biocontrol agent targets A. tumefaciens leucyl-tRNA synthetase (LeuRS), an essential enzyme for cell viability, while the agent itself survives by having a second, self-protective copy of the synthetase. In principle, this strategy from nature could be applied to other crop diseases by direct intervention.