Evidence for contemporary and historical gene flow between guppy populations in different watersheds, with a test for associations with adaptive traits.
In dendritic river systems, gene flow is expected to occur primarily within watersheds. Yet, rare cross-watershed transfers can also occur, whether mediated by (often historical) geological events or (often contemporary) human activities. We explored these events and their potential evolutionary consequences by analyzing patterns of neutral genetic variation (microsatellites) and adaptive phenotypic variation (male color) in wild guppies (Poecilia reticulata) distributed across two watersheds in northern Trinidad. We found the expected signatures of within-watershed gene flow; yet we also inferred at least two instances of cross-watershed gene flow-one in the upstream reaches and one further downstream. The upstream cross-watershed event appears to be very recent (41 ± 13 years), suggesting dispersal via recent flooding or undocumented human-mediated transport. The downstream cross-watershed event appears to be considerably older (577 ± 265 years), suggesting a role for rare geological or climatological events. Alongside these strong signatures of both contemporary and historical gene flow, we found little evidence of impacts on presumably adaptive phenotypic differentiation, except perhaps in the one instance of very recent cross-watershed gene flow. Selection in this system seems to overpower gene flow-at least on the spatiotemporal scales investigated here.


Shear-viscosity to entropy-density ratio of a relativistic hadron gas.
Ultrarelativistic heavy-ion collisions at the Brookhaven National Laboratory Relativistic Heavy Ion Collider (RHIC) are thought to have produced a state of matter called the quark-gluon plasma, characterized by a very small shear-viscosity to entropy-density ratio eta/s, near the lower bound predicted for that quantity by anti-de Sitter space/conformal field theory methods. As the produced matter expands and cools, it evolves through a phase described by a hadron gas with rapidly increasing eta/s. We calculate eta/s as a function of temperature in this phase both in and out of chemical equilibrium and find that its value poses a challenge for viscous relativistic hydrodynamics, which requires small values of eta/s in order to successfully describe the collective flow observables at the RHIC. We therefore conclude that the origin of the low viscosity matter at the RHIC must be in the partonic phase of the reaction.


Molecular and functional analysis of the mariner mutator element Mos1 in Drosophila.
The white-peach allele in Drosophila results from insertion of the transposable element mariner. The particular copy that is inserted in white-peach is an inactive copy referred to as the peach element. The peach element is excised at a high rate in the presence of active copies of mariner located elsewhere in the genome, and the excision of peach in somatic cells is recognized phenotypically by the occurrence of eye-color mosaicism in white-peach flies. Active mariner elements identified by their ability to induce high levels of white-peach mosaicism are denoted Mos (Mosaic) factors. We have sequenced and functionally analyzed the factor Mos1 originally identified in Drosophila mauritiana. The Mos1 element is 1286 base pairs in length, the same length as the peach element. It differs from the peach element in 11 nucleotide positions distributed throughout its length, including four amino acid replacements in the long open reading frame. Analysis of chimeric constructs between Mos1 and peach implies that functionally important differences occur in both the 5' and 3' halves of Mos1. A mariner element identical in sequence to Mos1 yields lower levels of mosaicism in transformants, implying that adjacent flanking sequences have important effects on Mos1 activity. Another mariner element, designated Ma351, isolated from a nonmosaic strain of D. mauritiana, differs from Mos1 in just three nucleotide positions. When introduced into the germline, Ma351 yields various levels of white-peach mosaicism depending on insertion site. These results imply that the activity of mariner elements is determined jointly by their own nucleotide sequences, by the effects of adjacent flanking sequences, and by longer-range position effects.


Geocoding Error, Spatial Uncertainty, and Implications for Exposure Assessment and Environmental Epidemiology.
Although environmental epidemiology studies often rely on geocoding procedures in the process of assigning spatial exposure estimates, geocoding methods are not commonly reported, nor are consequent errors in exposure assignment explored. Geocoding methods differ in accuracy, however, and, given the increasing refinement of available exposure models for air pollution and other exposures, geocoding error may account for an increasingly larger proportion of exposure misclassification. We used residential addresses from a reasonably large, dense dataset of asthma emergency department visits from all New York City hospitals (n = 21,183; 26.9 addresses/km2), and geocoded each using three methods (Address Point, Street Segment, Parcel Centroid). We compared missingness and spatial patterning therein, quantified distance and directional errors, and quantified impacts on pollution exposure estimates and assignment to Census areas for sociodemographic characterization. Parcel Centroids had the highest overall missingness rate (38.1%, Address Point = 9.6%, Street Segment = 6.1%), and spatial clustering in missingness was significant for all methods, though its spatial patterns differed. Street Segment geocodes had the largest mean distance error (µ = 29.2 (SD = 26.2) m; vs. µ = 15.9 (SD = 17.7) m for Parcel Centroids), and the strongest spatial patterns therein. We found substantial over- and under-estimation of pollution exposures, with greater error for higher pollutant concentrations, but minimal impact on Census area assignment. Finally, we developed surfaces of spatial patterns in errors in order to identify locations in the study area where exposures may be over-/under-estimated. Our observations provide insights towards refining geocoding methods for epidemiology, and suggest methods for quantifying and interpreting geocoding error with respect to exposure misclassification, towards understanding potential impacts on health effect estimates.


Apoptosis induced by doxorubicin in neurotumor cells is divorced from drug effects on ceramide accumulation and may involve cell cycle-dependent caspase activation.
Doxorubicin (0.5 microgram/ml) induced caspase-dependent apoptosis in SH-SY5Y neuroblastoma and CHP-100 neuroepithelioma cells. The apoptotic response started to be evident approximately 15 h after drug administration and, as monitored over a 48-h period, was more pronounced in CHP-100 than in SH-SY5Y cells. In both systems, apoptosis was accompanied by elevation of intracellular ceramide levels. Ceramide accumulation was blocked by the ceramide synthase inhibitor fumonisin B(1) (25 microM); this compound, however, did not prevent drug-induced apoptosis. Untreated cells from both lines expressed negligible p53 levels; on the other hand, whereas p53 and p21(Cip1/Waf1) were rapidly up-regulated in doxorubicin-treated SH-SY5Y cells, such a response was not observed in CHP-100 cells. Doxorubicin induced a G(2)/M phase block in both cell lines, but whereas the G(1) phase was markedly depleted in CHP-100 cells, it was substantially retained in SH-SY5Y cells. In the latter system, double G(1) and G(2)/M block largely preceded cell death; however, as apoptosis underwent completion, it selectively targeted late S and G(2)/M cells. Moreover, apoptosis suppression by caspase inhibition did not result in a recovery of the G(1) cell population. These results support the notion that doxorubicin-induced apoptosis and ceramide elevation are divorced events in neuroectodermal tumors and that p53 function is at least dispensable for apoptosis completion. Indeed, as G(1) cells appear to be refractory to doxorubicin-induced apoptosis, p53 up-regulation and p21(Cip1/Waf1) expression may provide an unfavorable setting for the apoptotic action of the drug.


Understanding leadership in community nursing in Scotland.
There is limited evidence concerning leadership in community nursing. NHS policy also fails to clarify and define what leadership is, though regarding it as key to developing safe and high quality care. This paper reports the findings of a research study that aimed to identify how leadership is perceived and experienced by community nurses, and to examine the interaction between recent policy and leadership development in community nursing. Mixed qualitative methods were used involving 31 individual interviews and three focus groups with community nurses and nurse leaders (n-39) in three health boards in Scotland. Findings indicate the leadership qualities valued by participants, including the importance of leaders' visibility. Team leaders in particular were recognised for their visibility and clinical leadership. Strategic and professional leadership was less evident, so acting as a barrier to the development of the profession. The strategic vision was often not clear to community nurses, and they engaged in differing ways with the strategies and action plans of senior nurse leaders. New leadership roles, like change, need time to evolve and new leaders need space and the education to develop leadership. Future leaders in community nursing need to focus beyond clinical leadership, ensuring that good leadership is a process requiring interdependence between leaders and followers.


Tubulin Polymerization Promoting Protein (TPPP) gene methylation and corpus callosum measures in maltreated children.
The goal of the current study was to evaluate the impact of Tubulin Polymerization Promoting Protein (TPPP) methylation on structural and fractional anisotropy (FA) corpus callosum (CC) measures. TPPP is involved in the development of white matter tracts in the brain and was implicated in stress-related psychiatric disorders in an unbiased whole epigenome methylation study. The cohort included 63 participants (11.73 y/o ±1.91) from a larger study investigating risk and resilience in maltreated children. Voxel-based morphometry (VBM) was used to process the structural data, fractional anisotropy (FA) was determined using an atlas-based approach, and DNA specimens were derived from saliva in two batches using the 450 K (N = 39) and 850 K (N = 24) Illumina arrays, with the data from each batch analyzed separately. After controlling for multiple comparisons and relevant covariates (e.g., demographics, brain volume, cell composition, 3 PCs), 850 K derived TPPP methylation values, in interaction with a dimensional measure of children's trauma experiences, predicted left and right CC body volumes and genu, body and splenium FA (p < .007, all comparisons). The findings in the splenium replicated in subjects with the 450 K data. The results extend prior investigations and suggest a role for TPPP in brain changes associated with stress-related psychiatric disorders.


Bioinformatics analysis of RNA-seq data revealed critical genes in colon adenocarcinoma.
RNA-seq data of colon adenocarcinoma (COAD) were analyzed with bioinformatics tools to discover critical genes in the disease. Relevant small molecule drugs, transcription factors (TFs) and microRNAs (miRNAs) were also investigated. RNA-seq data of COAD were downloaded from The Cancer Genome Atlas (TCGA). Differential analysis was performed with package edgeR. False positive discovery (FDR) < 0.05 and |log2 (fold change)|>1 were set as the cut-offs to screen out differentially expressed genes (DEGs). Gene coexpression network was constructed with package Ebcoexpress. GO enrichment analysis was performed for the DEGs in the gene coexpression network with DAVID. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was also performed for the genes with KOBASS 2.0. Modules were identified with MCODE of Cytoscape. Relevant small molecules drugs were predicted by Connectivity map. Relevant miRNAs and TFs were searched by WebGestalt. A total of 457 DEGs, including 255 up-regulated and 202 down-regulated genes, were identified from 437 COAD and 39 control samples. A gene coexpression network was constructed containing 40 DEGs and 101 edges. The genes were mainly associated with collagen fibril organization, extracellular matrix organization and translation. Two modules were identified from the gene coexpression network, which were implicated in muscle contraction and extracellular matrix organization, respectively. Several critical genes were disclosed, such as MYH11, COL5A2 and ribosomal proteins. Nine relevant small molecule drugs were identified, such as scriptaid and STOCK1N-35874. Accordingly, a total of 17 TFs and 10 miRNAs related to COAD were acquired, such as ETS2, NFAT, AP4, miR-124A, MiR-9, miR-96 and let-7. Several critical genes and relevant drugs, TFs and miRNAs were revealed in COAD. These findings could advance the understanding of the disease and benefit therapy development.


Cutting edge: NKG2D-dependent cytotoxicity is controlled by ligand distribution in the target cell membrane.
Although the importance of membrane microdomains in receptor-mediated activation of lymphocytes has been established, much less is known about the role of receptor ligand distribution on APC and target cells. Detergent-resistant membrane domains, into which GPI-linked proteins partition, are enriched in cholesterol and glycosphingolipids. ULBP1 is a GPI-linked ligand for natural cytotoxicity receptor NKG2D. To investigate how ULBP1 distribution on target cells affects NKG2D-dependent NK cell activation, we fused the extracellular domain of ULBP1 to the transmembrane domain of CD45. Introduction of this transmembrane domain eliminated the association of ULBP1 with the detergent-resistant membrane fraction and caused a significant reduction of cytotoxicity and degranulation by NK cells. Clustering and lateral diffusion of ULBP1 was not affected by changes in the membrane anchor. These results show that the partitioning of receptor ligands in discrete membrane domains of target cells is an important determinant of NK cell activation.


Cardiac Output Assessment in Patients Supported with Left Ventricular Assist Device: Discordance Between Thermodilution and Indirect Fick Cardiac Output Measurements.
Cardiac output (CO) assessed by thermodilution (TD) and indirect Fick (iFK) methods is commonly employed in left ventricular assist device (LVAD) patients; however, no study has assessed agreement. This study assesses correlation between these methods and association with hemodynamic/echocardiographic data in LVAD patients. Discordance was defined as a 20% difference between TD and iFK CO measurements. Bias and agreement were determined via the Bland-Altman technique in both the overall sample and iFK-stratified tertiles. Correlation with each assessment of CO and right heart catheterization (RHC) hemodynamics was performed. Among 111 RHCs, the mean CO for TD and iFK were 4.65 ± 1.33 (range: 1.44-9.30) and 5.37 ± 1.51 (range: 3.07-11.80) L/min (p < 0.001), respectively, with a calculated discordance of 45.9%. A correlation coefficient of 0.66 with a bias of -0.72 L/min was found. The lower and upper limit of precision were -3.12 and 1.68 L/min, respectively. By tertile analysis, bias (lower and upper limit of precision) for the low, middle, and high tertile groups were -0.24 (-1.88 and 1.40), -0.48 (-2.50 and 1.53), and -1.39 (-4.18 and 1.39) L/min, respectively. No significant correlation was found between either method with right atrial pressure or pulmonary capillary wedge pressure or any valvular condition. Substantial discrepancies exist between TD and iFK CO in LVAD patients. Although fixed bias was small, the limits of agreement extend into the clinically relevant area, with larger bias being present at higher CO. Studies with flow probes are needed to define which method better represents CO in LVAD patients.


The umbilical cord matrix is a better source of mesenchymal stem cells (MSC) than the umbilical cord blood.
Many studies have drawn attention to the emerging role of MSC (mesenchymal stem cells) as a promising population supporting new clinical concepts in cellular therapy. However, the sources from which these cells can be isolated are still under discussion. Whereas BM (bone marrow) is presented as the main source of MSC, despite the invasive procedure related to this source, the possibility of isolating sufficient numbers of these cells from UCB (umbilical cord blood) remains controversial. Here, we present the results of experiments aimed at isolating MSC from UCB, BM and UCM (umbilical cord matrix) using different methods of isolation and various culture media that summarize the main procedures and criteria reported in the literature. Whereas isolation of MSC were successful from BM (10:10) and (UCM) (8:8), only one cord blood sample (1:15) gave rise to MSC using various culture media [DMEM (Dulbecco's modified Eagle's medium) +5% platelet lysate, DMEM+10% FBS (fetal bovine serum), DMEM+10% human UCB serum, MSCGM] and different isolation methods [plastic adherence of total MNC (mononuclear cells), CD3+/CD19+/CD14+/CD38+-depleted MNC and CD133+- or LNGFR+-enriched MNC]. MSC from UCM and BM were able to differentiate into adipocytes, osteocytes and hepatocytes. The expansion potential was highest for MSC from UCM. The two cell populations had CD90+/CD73+/CD105+ phenotype with the additional expression of SSEA4 and LNGFR for BM MSC. These results clearly exclude UCB from the list of MSC sources for clinical use and propose instead UCM as a rich, non-invasive and abundant source of MSC.


Reliability and validity of the Turkish version of spinal cord lesion-related coping strategies.
Spinal Cord Lesion-Related Coping Strategies Questionnaire (SCL CSQ) is a specific test that is developed for evaluating the coping strategies of the persons with spinal cord injury (SCI). The aim of this study was to evaluate the reliability and validity of the Turkish version of SCL CSQ (SCL CSQ-T) in persons with SCI. One hundred persons with SCI were included in the study. All participants were evaluated with SCL CSQ-T and Brief Coping Styles Inventory (BCSI) at the baseline. SCL CSQ-T was repeated twice in 15 days. American Spinal Injury Association Impairment Scale and Functional Independence Measurement were used for the evaluation of the neurological loss severity and functional status. Emotional status was assessed by Hospital Anxiety and Depression Scale (HADS). Internal consistency reliability, test-retest reliability and construct validity of SCL CSQ-T were evaluated. Intraclass correlation coefficients of the SCL CSQ-T were between 0.51-0.86. Cronbach's alpha values and test-retest reliability of the acceptance, fighting spirit and social reliance subscales were good. Three factors were found in exploratory factor analysis. There was a positive correlation between the subscales of SCL CSQ-T. There was a statistically significant positive correlation between acceptance strategy of SCL CSQ-T and self-confidence and optimism of BCSI. The fighting spirit strategy positively correlated with self-confidence and optimistic strategies. The social reliance strategy positively correlated with helplessness and seeking social support. Coping strategies did not correlate with HADS. The results of this study revealed good internal consistency reliability, test-retest reliability and concurrent validity of the SCL CSQ-T factors acceptance and fighting spirit in relation to general coping strategies. The coping strategy social reliance needs revisions.


Congenital lung malformations : analysis of 27 cases.
Congenital lung malformations (CLM) include a complex range of developmental abnormalities. Currently, most are diagnosed prenatally or during early childhood. to investigate clinical and imaging findings of congenital lung malformations in children. Retrospective study of CLM diagnosed between 2000 and 2017 at the pediatric and neonatology department of Hedi Chaker Hospital. Analysis of clinical spectrum, diagnosis tools, and radiological appearances. Twenty seven cases of CLM have been investigated: 8 congenital lobar emphysema, 8 congenital cystic adenomatoid malformation, 8 pulmonary sequestrations, 2 bronchogenic cysts, and 1 hybrid lesion. Five (18,5%) patients were diagnosed antenatally and 22 (81,4%) postnatally. Symptoms occurred at a mean age of 9 months: Respiratory distress (11 cases), wheezing (4 cases), and pneumonia (7 cases). Antenatal ultrasound features include echogenic masses within the chest (2cases), the presence of cysts (2cases), unilateral pleural effusion (1 case) and hydramnios (2cases). A computed tomography scan was performed in all patients with a radio-histological concordance of 96%.


Pulse oximetry for perioperative monitoring.
Monitoring with pulse oximetry might improve patient outcome by enabling an early diagnosis and consequently, correction of perioperative events that might cause postoperative complications or even death. Only a few randomized clinical trials of pulse oximetry have been performed during anaesthesia, and in the recovery room which describe perioperative hypoxaemic events, postoperative cardiopulmonary complications and cognitive dysfunction. The objective of this review was to assess the effect of perioperative monitoring with pulse oximetry and to clearly identify the adverse outcomes that might be prevented or improved by the use of pulse oximetry. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2003, issue 1) MEDLINE (1966 to January 2003), EMBASE (1980 to January 2003), and by checking the reference lists of trials and review articles. All controlled trials that randomized patients to either pulse oximetry or no pulse oximetry during the perioperative period, including the operating and recovery room. Two reviewers independently assessed data in relation to events detectable by pulse oximetry, any serious complications that occurred during anaesthesia or in the postoperative period, intra- or postoperative mortality, and duration of recovery or intensive care stay. Formal statistical synthesis of individual trials was not performed in view of the variety of outcomes studied. Searching identified six reports. Four studies with data from a total of 21,773 patients were considered eligible for analysis. Only two studies specifically addressed the outcomes in question, both found no evidence of an effect on the rate of postoperative complications using perioperative pulse oximetry. Two studies used hypoxaemia detectable by pulse oximetry to assess the value of perioperative monitoring, although outcomes were not given. It was found that hypoxaemia was reduced in the pulse oximetry group both in the operating theatre and in the recovery room. During observation in the recovery room, the incidence of hypoxaemia in the pulse oximetry group was 1.5-3 times less. The postoperative cognitive function using the Wechsler memory scale and continuous reaction time was independent of perioperative monitoring with pulse oximetry. The other study showed that postoperative complications occurred in 10% of the patients in the oximetry group and in 9.4% in the control group. No statistically significant differences were detected in cardiovascular, respiratory, neurologic, or infectious complications in the two groups. The duration of hospital stay was a median of five days in both groups, and an equal number of in-hospital deaths was registered in the two groups. The studies confirmed that pulse oximetry can detect hypoxaemia and related events. However, we have found no evidence that pulse oximetry affects the outcome of anaesthesia. The conflicting subjective and objective results of the studies, despite an intense, methodical collection of data from a relatively large population, indicate that the value of perioperative monitoring with pulse oximetry is questionable in relation to improved reliable outcomes, effectiveness and efficiency.


[Fine structure of glial cells in the central nervous system of the tapeworm Grillotia erinaceus (Cestoda: Trypanorhyncha)].
The problem of glial cells existing in parasitic and free living flatworms is correlated with organization of parenchyma in platyhelmintes. In the contrary to the widespread opinion that myelin-like envelopes and glial cells do not exist in the nervous system of parasitic flatworms, it has been shown by ultrastructural researches that Amphilina foliacea (Cestoda, Amphilinidea) has well developed glial cells and myelin-like envelopes in the ganglia and main cords, which include both glial cells and intercellular components. The aim of our research was to reveal and investigate in details structural components corresponding to the concept of the glial cell in the CNS of Grillotia erinaceus (Cestoda: Trypanorhyncha). Three types of glial cells have been found. The first type is the fibroblast-like glial cells; cells locate in the cerebral ganglion, contain in cytoplasm and extract out fibrillar matrix, form desmosomes and have supporting function. The glial cells of the second type form myeline-like envelope of the giant axons and bulbar nerves in scolex and have laminar cytoplasm. These cells are numerous and exceed in number the neurons bodies into the nerve. The glial cells of the third type form multilayer envelopes in the main nerve cords; extra cellular fibers and gap-junctions take place between the layers. There are contacts between the glial cells of the third type and excretory epithelium but specialized contacts with neurons have been not found. The existing of glial cells in free living and parasitic flatworms is discussed.


Atherogenic Index of Plasma (AIP) a Tool to Assess Changes in Cardiovascular Disease Risk Post Laparoscopic Sleeve Gastrectomy.
Predictive indices like the atherogenic index of plasma (AIP) have been developed to estimate the risk of cardiovascular disease (CVD). Metabolic surgery is the most effective treatment for a rapid improvement of morbid obesity and its comorbidities such as type 2 diabetes (T2D) and CVD. A decreased reoccurrence of CVD after metabolic surgery has been reported by several studies. However, studies utilizing predictive indices for CVD risk in CVD-free morbid-obese patients who undertook laparoscopic sleeve gastrectomy (LSG) are lacking. Here, we use AIP as a tool to evaluate the improvement in CVD risk post-LSG in morbid-obese people who had no history of CVD. Method. We compared baseline, 6- and 12-month post-LSG score of AIP, vascular age, circulating biochemical markers related to CVD in two groups of BMI and age-matched morbid-obese participants with and without T2D. Results. At baseline, people with T2D had significantly higher AIP both, with morbid obesity (0.23 ± 0.06, p < 0.001) and normal weight (0.022 ± 0.05, p < 0.001) compared to their BMI-matched without T2D group. People with morbid obesity had low AIP (-0.083 ± 0.06). Vascular age was significantly higher in people with morbid obesity and T2D (65.8 ± 3.7year, p < 0.0001) compared to morbid obesity (37.9 ± 2.6 year). After one year, AIP was significantly reduced compared to baseline score in people with morbid obesity with/without T2D, respectively (-0.135 ± 0.07, p = 0.003; and -0.36 ± 0.04, p = 0.0002). Conclusion. Our data illuminates AIP as a reliable predictive index for CVD risk in morbid-obese people who had no history of CVD. Moreover, AIP accurately distinguishes between morbid obesity with T2D and morbid obesity and showed a rapid and significant reduction in CVD risk after LSG in people who had no history of CVD. This is a ClinicalTrials.gov registered trial (Reference NCT03038373).


Environmental exposure to polychlorinated biphenyls (PCBs) and breast cancer: a systematic review of the epidemiological evidence.
Some PCB congeners have shown oestrogenic effects, and this has raised concern that they may increase the risk of breast cancer. In this article we provide a quantitative review of the epidemiologic evidence on environmental exposure to PCBs and breast cancer risk. The vast majority of prospective and retrospective studies did not find any association between total PCB concentrations and breast cancer risk. No association was found for congeners in groups I (potentially oestrogenic) and III (biologically persistent phenobarbital-type cytochrome P450 inducers), according to the classification proposed by Wolff and Toniolo, while less consistent results were reported for group II (potentially anti-oestrogenic and immunotoxic, dioxin-like). Two studies found a threefold risk of postmenopausal breast cancer for women with an A2455G base change in exon 7 of the polymorphic CYP1A1 gene (a member of the cytochrome P450 family) and high PCB levels, compared with women with two wild-type alleles and low PCB, based however on very few cases. Thus, the epidemiological evidence does not support the hypothesis of an association of environmental exposure to PCBs in adulthood in the general population and risk of breast cancer, although uncertainties remain for selected subgroups of women or individual PCB congeners.


Effects of Cerebellothalamic Tractotomy on Cognitive and Emotional Functioning in Essential Tremor: A Preliminary Study in 5 Essential Tremor Patients.
Background: Subthalamic stereotactic interventions have recently caught renewed interest as a treatment for essential tremor (ET). However, it is not clear whether these interventions are associated with neurocognitive, mood or personality changes. Objective: To investigate neurocognition, neuropsychiatric functions and personality variables in patients with ET and to explore the neurocognitive and neuropsychiatric effects of cerebellothalamic tractotomy (CTT), a form of subthalamotomy. Methods: In our study, we investigated cognitive functions, frontal functions, mood and personality variables in 5 patients with intractable ET. Patients were tested before and 3 months after surgery using neuropsychological tests, clinical scales for depression, anxiety, anger regulation and a personality test. Results: Before surgery, ET patients showed normal neurocognitive function, a slightly elevated frontal lobe score in the dimensions mental control and memory, without being indicative of a frontal lesion, and no elevated depression or anxiety scores compared to norm values. After surgery, there was no change in neurocognitive function and no increase in depression or anxiety scores. Conclusion: In this exploratory study on 5 ET patients, CTT was not associated with alterations of mood or neurocognitive functions. © 2015 S. Karger AG, Basel.


Arterial viscoelasticity: role in the dependency of pulse wave velocity on heart rate in conduit arteries.
Experimental investigations have established that the stiffness of large arteries has a dependency on acute heart rate (HR) changes. However, the possible underlying mechanisms inherent in this HR dependency have not been well established. This study aimed to explore a plausible viscoelastic mechanism by which HR exerts an influence on arterial stiffness. A multisegment transmission line model of the human arterial tree incorporating fractional viscoelastic components in each segment was used to investigate the effect of varying fractional order parameter (α) of viscoelasticity on the dependence of aortic arch to femoral artery pulse wave velocity (afPWV) on HR. HR was varied from 60 to 100 beats/min at a fixed mean flow of 100 ml/s. PWV was calculated by intersecting tangent method (afPWVTan) and by phase velocity from the transfer function (afPWVTF) in the time and frequency domain, respectively. PWV was significantly and positively associated with HR for α ≥ 0.6; for α = 0.6, 0.8, and 1, HR-dependent changes in afPWVTan were 0.01 ± 0.02, 0.07 ± 0.04, and 0.22 ± 0.09 m/s per 5 beats/min; HR-dependent changes in afPWVTF were 0.02 ± 0.01, 0.12 ± 0.00, and 0.34 ± 0.01 m/s per 5 beats/min, respectively. This crosses the range of previous physiological studies where the dependence of PWV on HR was found to be between 0.08 and 0.10 m/s per 5 beats/min. Therefore, viscoelasticity of the arterial wall could contribute to mechanisms through which large artery stiffness changes with changing HR. Physiological studies are required to confirm this mechanism.NEW & NOTEWORTHY This study used a transmission line model to elucidate the role of arterial viscoelasticity in the dependency of pulse wave velocity on heart rate. The model uses fractional viscoelasticity concepts, which provided novel insights into arterial hemodynamics. This study also provides a means of assessing the clinical manifestation of the association of pulse wave velocity and heart rate.


Granulocytes do not have surface ABO antigens.
There is disagreement whether granulocytes have ABO antigens on their surfaces. We investigated this issue using pure populations of washed granulocytes from healthy individuals of known ABO, Lewis, and secretor phenotypes. The presence of ABO antigens on granulocytes was sought using four different techniques: a granulocyte-red cell mixed agglutination assay; a microleukoagglutination assay; a microgranulocyte cytotoxicity assay; and an immunoradiometric assay for granulocyte-associated IgG. Both fixed and unfixed granulocytes were tested. All tests were negative. Granulocytes do not have ABO antigens on their surfaces.


Critical Knowledge Gaps in Our Understanding of Environmental Cycling and Transmission of Leptospira spp.
Exposure to soil or water contaminated with the urine of Leptospira-infected animals is the most common way in which humans contract leptospirosis. Entire populations can be at high risk of leptospirosis while working in inundated fields, when engaging in aquatic sports, or after periods of heavy rainfall. The risk of infection after contact with these environmental sources depends on the ability of Leptospira bacteria to survive, persist, and infect new hosts. Multiple variables such as soil and water pH, temperature, and even environmental microbial communities are likely to shape the environmental conditions needed by the pathogen to persist. Here we review what is known about the environmental phase of the infectious Leptospira transmission cycle and identify knowledge gaps that will serve as a guide for future research.


Modulation of neuronal activity in the monkey putamen associated with changes in the habitual order of sequential movements.
The striatum, especially its dorsolateral part, plays a major role in motor skill learning and habit formation, but it is still unclear how this contribution might be mediated at the neuronal level. We recorded single neurons in the posterior putamen of two monkeys performing an overlearned sequence of arm reaching movements to examine whether task-related activities are sensitive to manipulations of the serial order of stimulus-target locations. The monkeys' capacity to learn sequential regularities was assessed by comparing arm movement latencies and saccadic ocular reactions when a fixed repeating sequence was replaced with a random sequence. We examined neurons classified as phasically active projection neurons (PANs) and tonically active presumed cholinergic interneurons (TANs). About one-third of the PANs (35/106, 33%) activated during specific parts of a trial displayed modulations of their level of activation when the sequential structure was changed. This differential activity consisted of either decreases or increases in activity without altering the time period during which task-related activations occurred. In addition, half of the TANs (41/80, 51%) changed their responses to task stimuli with the sequence switch, indicating that the response selectivity of TANs reflects the detection of the context that requires adaptation to changes in the serial order of stimulus presentations. Our findings suggest that task-related changes in activity of projection neurons may be an important factor contributing to the production and adjustment of sequential behavior executed in an automatic fashion, whereas putative interneurons may provide a signal for performance monitoring in specific contexts.


Comparison of phenotypic expression with genotypic transformation by using cloned, selectable markers.
The frequency of phenotypic expression of the herpes simplex virus type 1 tk and Escherichia coli gpt genes was compared with the frequency of genotypic transformation after calcium phosphate-mediated DNA transfection of a number of tk- and hprt- cell lines. In three of the five lines tested, the frequency of phenotypic expression was at most 10-fold higher than that of genotypic transformation as indicated by frequency of HAT resistance. The remaining two lines showed phenotypic responses which were 50- to 100-fold greater than the genotypic responses. The data indicate that the efficiency of DNA-mediated transformation with some cell lines can be limited by events after the uptake and expression of transfected DNA.


A clinical and economic assessment of adjuvanted trivalent versus standard egg-derived quadrivalent influenza vaccines among older adults in the United States during the 2018-19 and 2019-20 influenza seasons.
Clinical evidence supports use of enhanced influenza vaccines in older adults. Few economic outcome studies have compared adjuvanted trivalent inactivated (aIIV3) and standard egg-derived quadrivalent inactivated influenza vaccines (IIV4e). A retrospective cohort study was conducted leveraging deidentified US hospital data linked to claims data during the 2018-19 and 2019-20 influenza seasons. Relative vaccine effectiveness (rVE) was compared in adults aged ≥65 years receiving aIIV3 or IIV4e using inverse probability of treatment weighting (IPTW) and Poisson regression. An economic assessment quantified potential real-world cost savings. The study included 715,807 aIIV3 and 320,991 IIV4e recipients in the 2018-19 and 844,169 aIIV3 and 306,270 IIV4e recipients in the 2019-20 influenza seasons. aIIV3 was significantly more effective than IIV4e in preventing cardiorespiratory disease (2018-19 rVE = 6.2%; and 2019-20 rVE = 6.0%) and respiratory disease (2018-19 rVE = 8.9%; and 2019-20 rVE = 10.1%). During the 2018-19 influenza season cardiorespiratory hospitalization cost savings for the aIIV3 population was $392 M, and $221 M for the 2019-20 season. Respiratory hospitalization cost savings for the aIIV3 population was $145 M and $97 M, respectively. Our findings suggest that aIIV3 provides clinical and economic advantages versus IIV4e in the elderly.


Roughness Analysis on Composite Materials (Microfilled, Nanofilled and Silorane) After Different Finishing and Polishing Procedures.
The finishing and polishing of composite materials affect the restoration lifespan. The market shows a variety of finishing and polishing procedures and the choice among them is conditioned by different factors such as the resulting surface roughness. In the present study, 156 samples were realized with three composite materials, -microfilled, nanofilled and silorane-, and treated with different finishing and polishing procedures. Profilometric analyses were carried out on the samples' surface, the measured roughness values were submitted to statistical analysis. A complete factorial plan was drawn up and two-way analysis of variance (ANOVA) was carried out to investigate whether the following factors affect the values of roughness: (i) material; (ii) polishing/finishing procedure. Tukey post-hoc test was also conducted to evaluate any statistically significant differences between the material/procedure combinations. The results show that the tested materials do not affect the resulting surface quality but roughness values depend on the finishing/polishing procedure adopted. The procedures that involve: (a) the finishing with medium Sof-Lex discs and (b) the finishing with two tungsten carbide multi-blade milling cutters Q series and UF series are those that allow the lowest values of roughness to be obtained.


False-positive results in a recombinant severe acute respiratory syndrome-associated coronavirus (SARS-CoV) nucleocapsid enzyme-linked immunosorbent assay due to HCoV-OC43 and HCoV-229E rectified by Western blotting with recombinant SARS-CoV spike polypeptide.
Using paired serum samples obtained from patients with illness associated with increases in anti-human coronavirus OC43 (HCoV-OC43) or anti-HCoV-229E antibodies, we examined the possibility of false-positive results detected in a recombinant severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) nucleocapsid protein immunoglobulin G enzyme-linked immunosorbent assay (ELISA). Three of the 21 and 1 of the 7 convalescent-phase serum samples from persons with increases in antibodies against HCoV-OC43 and HCoV-229E, respectively, tested positive by the recombinant SARS-CoV nucleocapsid protein-based ELISA. None of these samples were found to contain a specific antibody in the recombinant SARS-CoV spike polypeptide-based Western blot assay.


Effects of Antigravity Treadmill Training on Gait, Balance, and Fall Risk in Children With Diplegic Cerebral Palsy.
The aim of this study was to investigate the effects of antigravity treadmill training on gait, balance, and fall risk in children with diplegic cerebral palsy. Thirty children with diplegic cerebral palsy were selected for this randomized controlled study. They were randomly assigned to (1) an experimental group that received antigravity treadmill training (20 mins/d, 3 d/wk) together with traditional physical therapy for 3 successive mos and (2) a control group that received only traditional physical therapy program for the same period. Outcomes included selected gait parameters, postural stability, and fall risk. Outcomes were measured at baseline and after 3 mos of intervention. Children in both groups showed significant improvements in the mean values of all measured variables (P < 0.05), with significantly greater improvements in the experimental group than the control group. The posttreatment gait parameters (i.e., velocity, stride length, cadence, and percent of time spent in double-limb support) were 0.74 m/sec, 119 steps/min, 0.75 m/sec, 0.65 sec, and 55.9% as well as 0.5 m, 125 steps/min, 0.6 m/sec, 0.49 sec, and 50.4% for the experimental and control group, respectively. Antigravity treadmill training may be a useful tool for improving gait parameters, balance, and fall risk in children with diplegic cerebral palsy.


Percutaneous sonography-guided radiofrequency ablation in the management of parathyroid adenoma.
Parathyroid adenoma is a major cause of primary hyperparathyroidism. Treatment usually involves the surgical removal of one or more parathyroid glands. However, specific localising techniques have boosted the development of nonsurgical, minimally invasive procedures, such as percutaneous radiofrequency ablation (RFA) under ultrasonographic guidance, which are gaining popularity as a method of treatment. This paper reports two male patients who presented with hypercalcaemia (highest serum calcium level 4.56 mmol/L and 4.57 mmol/L, respectively) and hyperparathyroidism (highest serum parathyroid hormone [PTH] level 772 ng/L and > 1,900 ng/L, respectively) due to solitary parathyroid adenoma. Four days after percutaneous RFA, the serum calcium levels in both patients decreased and PTH levels showed a significant decreasing trend. Both patients recovered well with normal levels of calcium and improvement of symptoms. Thus, we conclude that RFA may be a safe and effective therapeutic option in the treatment of parathyroid adenoma.


The genetics of resistance to lettuce drop (Sclerotinia spp.) in lettuce in a recombinant inbred line population from Reine des Glaces × Eruption.
Two QTLs for resistance to lettuce drop, qLDR1.1 and qLDR5.1, were identified. Associated SNPs will be useful in breeding for lettuce drop and provide the foundation for future molecular analysis. Lettuce drop, caused by Sclerotinia minor and S. sclerotiorum, is an economically important disease of lettuce. The association of resistance to lettuce drop with the commercially undesirable trait of fast bolting has hindered the integration of host resistance in control of this disease. Eruption is a slow-bolting cultivar that exhibits a high level of resistance to lettuce drop. Eruption also is completely resistant to Verticillium wilt caused by race 1 of Verticillium dahliae. A recombinant inbred line population from the cross Reine des Glaces × Eruption was genotyped by sequencing and evaluated for lettuce drop and bolting in separate fields infested with either S. minor or V. dahliae. Two quantitative trait loci (QTLs) for lettuce drop resistance were consistently detected in at least two experiments, and two other QTLs were identified in another experiment; the alleles for resistance at all four QTLs originated from Eruption. A QTL for lettuce drop resistance on linkage group (LG) 5, qLDR5.1, was consistently detected in all experiments and explained 11 to 25% of phenotypic variation. On LG1, qLDR1.1 was detected in two experiments explaining 9 to 12% of the phenotypic variation. Three out of four resistance QTLs are distinct from QTLs for bolting; qLDR5.1 is pleiotropic or closely linked with a QTL for early bolting; however, the rate of bolting shows only a small effect on the variance in resistance observed at this locus. The SNP markers linked with these QTLs will be useful in breeding for resistance through marker-assisted selection.


Shear bond strength of Dyract compomer material to dentin of primary molars.
The purpose of this study was to measure and compare the shear bond strength and fracture pattern of Dyract compomer material with and without use of PSA prime/adhesive as well as 35% phosphoric acid etching to the buccal dentin of primary first and second molars. In addition, micromorphology of the restorative surfaces opposing the tooth structure following these different surface treatments was evaluated. For shear bond strength measurement and fracture pattern evaluation, 36 extracted non-restored human primary molars with mild to moderate caries divided into 3 groups of 12 teeth each were used. Dyract with and without use of PSA prime/adhesive as well as 35% phosphoric acid etching for 15 seconds prior to placement of PSA prime/adhesive was applied to the buccal surface of exposed dentin. A standardized tube of Dyract was placed on each dentin surface and polymerized. The tubes were sheared off with a Universal testing machine at a cross head speed of 12.7 mm/min. For evaluation of the restorative surfaces opposing the tooth structure, 9 teeth divided into 3 groups of 3 teeth each were used to prepare the specimens, which were then demineralized in 10% hydrochloric acid for 24 hours. Fitting surfaces of these specimens were prepared and examined using scanning electron microscope. Tukey's multiple range test showed that the shear bond strength of Dyract with PSA prime/adhesive (group 1) was statistically significantly higher than Dyract without PSA prime/adhesive (group 2) and phosphoric acid etching (group 3). The shear bond strength in group 1 averaged 5.89 +/- 1.40 (X + SD MPa) while for groups 2 and averaged 1.49 +/- 0.69 and 3.69 +/- 0.89 respectively. Pretreatment of dentin surface with 35% phosphoric acid increased resin tags formation but it did significantly lower shear bond strength of Dyract with PSA prime/adhesive to dentin of primary molars. Bond failure patterns for all groups were only adhesive and mixed type failures.


Bayesian safety surveillance with adaptive bias correction.
Postmarket safety surveillance is an integral part of mass vaccination programs. Typically relying on sequential analysis of real-world health data as they accrue, safety surveillance is challenged by sequential multiple testing and by biases induced by residual confounding in observational data. The current standard approach based on the maximized sequential probability ratio test (MaxSPRT) fails to satisfactorily address these practical challenges and it remains a rigid framework that requires prespecification of the surveillance schedule. We develop an alternative Bayesian surveillance procedure that addresses both aforementioned challenges using a more flexible framework. To mitigate bias, we jointly analyze a large set of negative control outcomes that are adverse events with no known association with the vaccines in order to inform an empirical bias distribution, which we then incorporate into estimating the effect of vaccine exposure on the adverse event of interest through a Bayesian hierarchical model. To address multiple testing and improve on flexibility, at each analysis timepoint, we update a posterior probability in favor of the alternative hypothesis that vaccination induces higher risks of adverse events, and then use it for sequential detection of safety signals. Through an empirical evaluation using six US observational healthcare databases covering more than 360 million patients, we benchmark the proposed procedure against MaxSPRT on testing errors and estimation accuracy, under two epidemiological designs, the historical comparator and the self-controlled case series. We demonstrate that our procedure substantially reduces Type 1 error rates, maintains high statistical power and fast signal detection, and provides considerably more accurate estimation than MaxSPRT. Given the extensiveness of the empirical study which yields more than 7 million sets of results, we present all results in a public R ShinyApp. As an effort to promote open science, we provide full implementation of our method in the open-source R package EvidenceSynthesis.


[Prevalence and detection of depressive disorders in primary care].
To study the prevalence of depression in primary care, the detection of depressive disorders by primary care physicians, and the factors that influence detection. Cross-sectional, descriptive study. Gavà II Primary Care Center, in Barcelona, Spain. A total of 400 people between the ages of 18 and 65 years were chosen randomly from among those who attended appointments with their primary care physician. A sociodemographic questionnaire and the Beck Depression Inventory (BDI) screening test were administered, and the participant s medical record was reviewed. In a subsample of 40 participants, the Mini-International Neuropsychiatric Interview (MINI) was also administered. The optimum cutoff score for the BDI was estimated with reference to the MINI results. A cutoff score of 20/21 for the BDI had a sensitivity of 86.7% and a specificity of 92%, when the MINI score was used as a reference. The adjusted prevalence of depressive disorder in our primary care setting was 20.2% overall, 8.1% in men, and 26.8% in women (odds ratio 4.15, p < 0.01). The physician detected depressive symptoms in 55.7% of all likely cases of depression. Persons who scored >= 21 on the BDI made more visits to their primary care physician, and had more stressful life events, than those who scored 20. The prevalence of depression in our primary care setting is high. The disorder was underdiagnosed in as many as 44.3% of the persons likely to have depressive disorder (especially women, widows and widowers, retired persons, persons who had experienced stressful life events, and frequent users of primary care services).


Differentiation of Purkinje fibres and ordinary ventricular and atrial myocytes in the bovine heart: an immuno- and enzyme histochemical study.
The differentiation of Purkinje fibres and ordinary ventricular and atrial myocytes in bovine hearts was studied with specific antibodies against M-line proteins (MM-creatine kinase and myomesin) and with enzyme histochemistry (succinate dehydrogenase and mitochondrial glycerol-3-phosphate dehydrogenase). MM-creatine kinase was detected at an earlier stage in Purkinje fibres and atrial myocytes than in ordinary ventricular myocytes. The findings are in agreement with previous ultrastructural observations that an earlier appearance of a dense M-band occurs in Purkinje fibres than in ordinary ventricular myocytes. Myomesin was detected in all three cell types even at early foetal stages, in accordance with suggestions that it is an integral component of the myofibrillar structure. The activity of succinate dehydrogenase gradually increased in both ordinary ventricular and atrial myocytes, while the activity of mitochondrial glycerol-3-phosphate dehydrogenase was high at different stages of early foetal development in the two tissues, finally becoming low in the adult stage. The activity of succinate dehydrogenase and mitochondrial glycerol-3-phosphate dehydrogenase seemed to remain unchanged in the Purkinje fibres from early to late foetal stages. The present study shows that the Purkinje fibres are already different from ordinary ventricular myocytes at early foetal stages and that the two cell types differentiate in different ways. It is concluded that there are also developmental differences between ordinary ventricular and atrial myocytes.


RatLesNetv2: A Fully Convolutional Network for Rodent Brain Lesion Segmentation.
We present a fully convolutional neural network (ConvNet), named RatLesNetv2, for segmenting lesions in rodent magnetic resonance (MR) brain images. RatLesNetv2 architecture resembles an autoencoder and it incorporates residual blocks that facilitate its optimization. RatLesNetv2 is trained end to end on three-dimensional images and it requires no preprocessing. We evaluated RatLesNetv2 on an exceptionally large dataset composed of 916 T2-weighted rat brain MRI scans of 671 rats at nine different lesion stages that were used to study focal cerebral ischemia for drug development. In addition, we compared its performance with three other ConvNets specifically designed for medical image segmentation. RatLesNetv2 obtained similar to higher Dice coefficient values than the other ConvNets and it produced much more realistic and compact segmentations with notably fewer holes and lower Hausdorff distance. The Dice scores of RatLesNetv2 segmentations also exceeded inter-rater agreement of manual segmentations. In conclusion, RatLesNetv2 could be used for automated lesion segmentation, reducing human workload and improving reproducibility. RatLesNetv2 is publicly available at https://github.com/jmlipman/RatLesNetv2.


Transcriptional modulation of TCR, Notch and Wnt signaling pathways in SEB-anergized CD4+ T cells.
Gene expression changes in CD4 + Vbeta8+ T cells energized by in vivo exposure to staphylococcal enterotoxin B (SEB) bacterial superantigen compared to CD4 + Vbeta8+ non-energic T cells were assessed using DNA microarrays containing 5184 murine complementary DNAs. Anergy in splenic T cells of SEB-immunized BALB/c mice was verified by dramatically reduced proliferative capacity and an 8 x overexpression of GRAIL mRNA in CD4 + Vbeta8+ T cells taken from mice 7 days after injection. At an Associative t-test threshold of P<0.0005, 96 genes were overexpressed or detected only in anergic T cells, while 256 genes were suppressed or not detected in anergic T cells. Six of eight differential expressions tested using real-time quantitative PCR were validated. Message for B-Raf was detected only in non-anergic cells, while expression of the TCR signaling modulator Slap (Src-like adapter protein) and the TCR zeta-chain specific phosphatase Ptpn3 was enhanced. Modulation of multiple genes suggests downregulation of Wnt/beta-catenin signaling and enhanced Notch signaling in the anergic cells. Consistent with previous reports in a non-superantigen in vivo anergy model, mRNA for CD18 and the transcription factor Satb1 (special AT-rich-binding protein 1) was increased in SEB-energized T cells. This is the first report of global transcriptional changes in CD4+ T cells made anergic by superantigen exposure.


Utilizing protein structure to identify non-random somatic mutations.
Human cancer is caused by the accumulation of somatic mutations in tumor suppressors and oncogenes within the genome. In the case of oncogenes, recent theory suggests that there are only a few key "driver" mutations responsible for tumorigenesis. As there have been significant pharmacological successes in developing drugs that treat cancers that carry these driver mutations, several methods that rely on mutational clustering have been developed to identify them. However, these methods consider proteins as a single strand without taking their spatial structures into account. We propose an extension to current methodology that incorporates protein tertiary structure in order to increase our power when identifying mutation clustering. We have developed iPAC (identification of Protein Amino acid Clustering), an algorithm that identifies non-random somatic mutations in proteins while taking into account the three dimensional protein structure. By using the tertiary information, we are able to detect both novel clusters in proteins that are known to exhibit mutation clustering as well as identify clusters in proteins without evidence of clustering based on existing methods. For example, by combining the data in the Protein Data Bank (PDB) and the Catalogue of Somatic Mutations in Cancer, our algorithm identifies new mutational clusters in well known cancer proteins such as KRAS and PI3KC α. Further, by utilizing the tertiary structure, our algorithm also identifies clusters in EGFR, EIF2AK2, and other proteins that are not identified by current methodology. The R package is available at: http://www.bioconductor.org/packages/2.12/bioc/html/iPAC.html. Our algorithm extends the current methodology to identify oncogenic activating driver mutations by utilizing tertiary protein structure when identifying nonrandom somatic residue mutation clusters.


Batch effects in single-cell RNA-sequencing data are corrected by matching mutual nearest neighbors.
Large-scale single-cell RNA sequencing (scRNA-seq) data sets that are produced in different laboratories and at different times contain batch effects that may compromise the integration and interpretation of the data. Existing scRNA-seq analysis methods incorrectly assume that the composition of cell populations is either known or identical across batches. We present a strategy for batch correction based on the detection of mutual nearest neighbors (MNNs) in the high-dimensional expression space. Our approach does not rely on predefined or equal population compositions across batches; instead, it requires only that a subset of the population be shared between batches. We demonstrate the superiority of our approach compared with existing methods by using both simulated and real scRNA-seq data sets. Using multiple droplet-based scRNA-seq data sets, we demonstrate that our MNN batch-effect-correction method can be scaled to large numbers of cells.


Transanal Irrigation in the Treatment of Children With Intractable Functional Constipation.
The aim of the study was to assess the treatment efficacy of transanal irrigation and parental satisfaction in children with intractable functional constipation (FC) treated with Peristeen. Cross-sectional survey study among parents of children (age 0-18 years) treated with Peristeen for FC (based on the Rome III criteria). Anonymous questionnaires were sent out to parents via mail, these consisted of 25 self-developed, multiple-choice questions regarding the use of Peristeen, current gastrointestinal symptoms, adverse effects of Peristeen, concomitant medication use, and parental satisfaction. Out of 91 invited families, 67 (74%) returned the questionnaire. In total, 84% of patients experienced fecal incontinence prior to treatment. Out of all children who still used Peristeen at the time of survey (n = 49), fecal incontinence had resolved completely in 41%, 12% experienced occasional episodes of fecal incontinence (<1 episode per week) and the remaining 47% still experienced episodes of fecal incontinence regularly (≥1 time per week). A total of 28 children (42%) experienced pain during rectal irrigation, especially during insertion of the catheter, inflating the balloon, or during irrigation. Overall, 86% of the parents were satisfied with the result of transanal irrigation and 67% reported that they would continue using transanal irrigation for the treatment of their child's symptoms. Transanal irrigation may be effective in the treatment of children with FC and renders a high parental satisfaction. Future prospective studies, preferably RCTs, are necessary to further evaluate this treatment option.


Induction of apoptosis in colorectal carcinoma cells treated with 4-hydroxy-2-nonenal and structurally related aldehydic products of lipid peroxidation.
The oxidation of polyunsaturated fatty acids during oxidative stress gives rise to a series of toxic alpha,beta-unsaturated aldehydes, including the electrophile 4-hydroxy-2-nonenal (4-HNE) and the related aldehydes, 4-hydroperoxy-2-nonenal (4-HPNE) and 4-oxo-2-nonenal (4-ONE). We synthesized these compounds, as well as the resolved enantiomers of 4-HNE, and compared their toxicities and apoptotic responses in the human colorectal cancer cell line RKO. All of these molecules execute similar death responses at comparable doses over almost identical time frames in RKO cells. The apoptotic response induced by 4-HPNE, 4-ONE, and 4-HNE enantiomers involves activation of caspases, proteolysis of downstream caspase targets, and nucleosomal DNA fragmentation. The results presented herein suggest that these molecules commonly activate certain signaling pathways that control cell death irrespective of their reactive properties.


Rat lung glutathione release: response to oxidative stress and selenium deficiency.
We performed experiments to characterize the glutathione-dependent metabolism occurring during tert-butyl hydroperoxide infusion in isolated perfused rat lungs and to examine the effect of selenium deficiency on this metabolism. Selenium deficiency resulted in decreased lung glutathione peroxidase activity but normal glutathione reductase activity and glutathione content. Infusion of the hydroperoxide into control lungs caused a proportional increase in tissue glutathione disulfide (GSSG) concentration and release of GSSG into the perfusate up to an infusion rate of 250 nmol of tert-butyl hydroperoxide X min-1 X 100 g body wt-1. Infusion rates greater than this resulted in continued rise of tissue GSSG concentrations but GSSG release into the perfusate plateaued. Infusion of tert-butyl hydroperoxide into selenium-deficient rat lungs resulted in much lower concentrations of tissue GSSG and GSSG release into the perfusate; however, release in the selenium-deficient rat lung was also found to be saturable at infusion rates of 450 nmol of tert-butyl hydroperoxide X min-1 X 100 g of body wt-1. Selenium deficiency in the rat decreases the rate of reduction of infused tert-butyl hydroperoxide by glutathione and may predispose the lung to free radical damage.


A NIR-driven green affording-oxygen microrobot for targeted photodynamic therapy of tumors.
Photodynamic therapy (PDT) is a light-activated local treatment modality that has promising potential in cancer therapy. However, ineffective delivery of photosensitizers and hypoxia in the tumor microenvironment severely restrict the therapeutic efficacy of PDT. Herein, phototactic Chlorella (C) is utilized to carry photosensitizer-encapsulated nanoparticles to develop a near-infrared (NIR) driven green affording-oxygen microrobot system (CurNPs-C) for enhanced PDT. Photosensitizer (curcumin, Cur) loaded nanoparticles are first synthesized and then covalently attached to C through amide bonds. An in vitro study demonstrates that the developed CurNPs-C exhibits continuous oxygen generation and desirable phototaxis under NIR treatment. After intravenous injection, the initial 660 nm laser irradiation successfully induces the active migration of CurNPs-C to tumor sites for higher accumulation. Upon the second 660 nm laser treatment, CurNPs-C produces abundant oxygen, which in turn induces the natural product Cur to generate more reactive oxygen species (ROS) that significantly inhibit the growth of tumors in 4T1 tumor-bearing mice. This contribution showcases the ability of a light-driven green affording-oxygen microrobot to exhibit targeting capacity and O2 generation for enhancing photodynamic therapy.


[Results of the biochemical examination of persons who were in contact with infectious hepatitis patients].
The results or biochemical examination (by the micromethod) of persons having close contacts with infectious hepatitis patients and persons living outside the foci of this infection in one of the regions of the Uzbek SSR with unfavorable situation in respect of infectious hepatitis are presented. Among the first group of persons those with hyperenzymemia constituted 2%. Outside the foci of infection such persons constituted 1.5%. The ratio of the frequency of the laboratory diagnosis of the disease to the number of officially registered cases was 0.1:1.0. The greatest percentage of persons with biochemical shifts was established in October-December (2.2-3.4) and among the patients examined after the 20th day of hospitalization (2.6 ), as well as among children under 7 years (3.2) and in the age group 20-29 years (2.6).


Host factors associated with the Sindbis virus RNA-dependent RNA polymerase: role for G3BP1 and G3BP2 in virus replication.
Sindbis virus (SINV) is the prototype member of the Alphavirus genus, whose members cause severe human diseases for which there is no specific treatment. To ascertain host factors important in the replication of the SINV RNA genome, we generated a SINV expressing nsP4, the viral RNA-dependent RNA polymerase, with an in-frame 3xFlag epitope tag. Proteomic analysis of nsP4-containing complexes isolated from cells infected with the tagged virus revealed 29 associated host proteins. Of these, 10 proteins were associated only at a later time of infection (12 h), 14 were associated both early and late, and five were isolated only at the earlier time (6 h postinfection). These results demonstrate the dynamic nature of the virus-host interaction that occurs over the course of infection and suggest that different host proteins may be required for the multiple functions carried out by nsP4. Two related proteins found in association with nsP4 at both times of infection, GTPase-activating protein (SH3 domain) binding protein 1 (G3BP1) and G3BP2 were also previously identified as associated with SINV nsP2 and nsP3. We demonstrate a likely overlapping role for these host factors in limiting SINV replication events. The present study also identifies 10 host factors associated with nsP4 6 h after infection that were not found to be associated with nsP2 or nsP3. These factors are candidates for playing important roles in the RNA replication process. Identifying host factors essential for replication should lead to new strategies to interrupt alphavirus replication.


Spanish Validation of the Internet Gaming Disorder Scale-Short Form (IGDS9-SF): Prevalence and Relationship with Online Gambling and Quality of Life.
Online gaming is a very common form of leisure among adolescents and young people, although its excessive and/or compulsive use is associated with psychological impairments in a minority of gamers. The latest (fifth) edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5, Section III) tentatively introduced Internet Gaming Disorder (IGD). Since then, a number of evaluation tools using the DSM-5 criteria have been developed, including the Internet Gaming Disorder Scale-Short Form (IGDS9-SF). The main objective of this study was to translate and adapt the IGDS9-SF into Spanish, as well as to obtain indicators relating to its validity and reliability. The Spanish version of four scales were administered: IGDS9-SF, Mobile Phone-Related Experiences Questionnaire (CERM), Online Gambling Disorder Questionnaire (OGD-Q), and KIDSCREEN-27. The sample comprised 535 Vocational Training students (mean age 18.35 years; SD±2.13; 78.5% males) who reported playing video games in the past 12 months. Confirmatory factor analysis yielded a one-dimensional model with a good fit while the reliability indicators were satisfactory. Findings indicated that 1.9% of gamers were classified with IGD (meeting five or more criteria for more than 12 months). Additionally, another 1.9% were considered gamers 'at-risk' because they endorsed four criteria. Positive and significant relationships were found between the IGDS9-SF, the CERM, and the OGD-Q. Participants classified with IGD had poorer health-related quality of life. In conclusion, the Spanish IGDS9-SF is a valid and reliable instrument to assess IGD according to the DSM-5.


A New Version of the Tissue Composition-Based Model for Improving the Mechanism-Based Prediction of Volume of Distribution at Steady-State for Neutral Drugs.
In-vitro models are available in the literature for predicting the volume of distribution at steady-state (Vdss) of drugs. The mechanistic model refers to the tissue composition-based model (TCM), which includes important factors that govern Vdss such as drug physiochemistry and physiological data. The recognized TCM published by Rodgers and Rowland (TCM-RR) and a subsequent adjustment made by Simulations Plus Inc. (TCM-SP) have been shown to be generally less accurate with neutral compared to ionized drugs. Therefore, improving these models for neutral drugs becomes necessary. The objective of this study was to propose a new TCM for improving the prediction of Vdss for neutral drugs. The new TCM included two modifications of the published models (i) accentuate the effect of the blood-to-plasma ratio (BPR) that should cover permeated molecules across the biomembranes, which is lacking in these models for neutral compounds, and (ii) use a different approach to estimate the binding in tissues. The new TCM was validated with a large dataset of 202 commercial and proprietary compounds including preclinical and clinical data. All scenario datasets were predicted more accurately with the TCM-New, whereas all statistical parameters indicate that the TCM-New showed significant improvements in terms of accuracy over the TCM-RR and TCM-SP. Predictions of Vdss were frequently more accurate for the TCM-new with 83% within twofold error versus only 50% for the TCM-RR. And more than 95% of the predictions were within threefold error and patient interindividual differences can be predicted with the TCM-New, greatly exceeding the accuracy of the published models. Overall, the new TCM incorporating BPR significantly improved the Vdss predictions in animals and humans for neutral drugs, and, hence, has the potential to better support the drug discovery and facilitate the first-in-human predictions.


Deciphering the mechanisms of reciprocal regulation or interdependence at the cannabinoid CB1 receptors and cyclooxygenase-2 level: Effects on mood, cognitive implications, and synaptic signaling.
The lipid endocannabinoid system refers to endogenous cannabinoids (eCBs), the enzymes involved in their synthesis and metabolism, and the G protein-coupled cannabinoid receptors (GPCRs), CB1, and CB2. CB1 receptors (CB1Rs) are distributed in the brain at presynaptic terminals. Their activation induces inhibition of neurotransmitter release, which are gamma-aminobutyric acid (GABA), glutamate (Glu), dopamine, norepinephrine, serotonin, and acetylcholine. Postsynaptically localized CB1Rs regulate the activity of selected ion channels and N-methyl-D-aspartate receptors (NMDARs). CB2Rs are mainly peripheral and will not be considered here. Anandamide metabolism, mediated by cyclooxygenase-2 (COX-2), generates anandamide-derived prostanoids. In addition, COX-2 regulates the formation of CB1 ligands, which reduce excitatory transmission in the hippocampus (HC). The role of CB1Rs and COX-2 has been described in anxiety, depression, and cognition, among other central nervous system (CNS) disorders, affecting neurotransmission and behavior of the synapses. This review will analyze common pathways, mechanisms, and behavioral effects of manipulation at the CB1Rs/COX-2 level.


The effect of storage at - 80 degrees C on the activities of cytoplasmic, mitochondrial and microsomal enzymes in rat liver.
The effect of storage at - 80 degrees C for 1-28 days on the activity of 12 enzymes in intact liver tissue, liver extract and isolated hepatic microsomes was investigated. To find optimal conditions for tissue homogenization for this study the effects of three types of homogenization on the activity of 10 enzymes from different cell compartments were compared. The activities of glucokinase and phosphofructokinase decreased markedly during storage of both supernatant and liver tissue. Storage of liver tissue increased the activity of mitochondrial enzymes or isoenzymes. While this effect can be explained by additional disintegration of liver tissue caused by freezing and thawing for enzymes like glutamine dehydrogenase, other mechanisms may be involved in the prolonged increase observed in the activity of citrate synthase and xanthine oxidase during storage. The activity of a number of enzymes from the cytosol, mitochondria and microsomes decreased more markedly in the stored liver samples than in the stored supernatant or in the stored microsomal pellet. Cytochrome P 450 content remained stable throughout the whole storage period in both intact liver tissue and isolated microsomes.


Early development of defibrillation devices.
The recognition of ventricular fibrillation and the development of defibrillation devices are recounted, and recent advances are described. Early experiences with and studies of cardiac arrest are discussed. Transthoracic defibrillation, which was first applied successfully in 1947, is reviewed, and significant research is summarized. The development and present state of the implanted defibrillator, which was suggested in the late 1960s, are examined.


Mortality from sexually transmitted diseases in reproductive-aged women: United States, 1999-2010.
I estimated the sexually transmitted disease-related mortality among US reproductive-aged women from 1999 to 2010. I estimated mortality from National Center for Health Statistics' Multiple Cause of Death data. I defined reproductive age as 15 to 44 years. For diseases partially caused by sexual transmission, I estimated the proportion attributable to sexual transmission from the literature. To calculate mortality rates, I estimated number of deaths from each disease and Census Bureau population for reproductive-aged women for 1999 to 2010. From 1999 to 2010, the cumulative sexually transmitted disease-related mortality rate decreased by 49%, from 5.3 to 2.7 deaths per 100 000. The primary contributors were HIV and human papilloma virus infections. Mortality from sexually transmitted HIV infection decreased by 62%, from 3.4 to 1.3 deaths per 100 000. Mortality from human papilloma virus-associated gynecologic cancers decreased by 19%, from 1.6 deaths per 100 000 in 1999 to 1.3 deaths per 100 000 in 2010. Screening and treatment for sexually transmitted diseases may reduce mortality. Research is needed to determine whether sexually transmitted disease-related morbidity among reproductive-aged women has decreased over the past decade.


Satellite DNA derived from 5S rDNA in Physalaemus cuvieri (Anura, Leiuperidae).
In the present study, we describe for the first time a family of 190-bp satellite DNA related to 5S rDNA in anurans and the existence of 2 forms of 5S rDNA, type I (201 bp) and type II (690 bp). The sequences were obtained from genomic DNA of Physalaemus cuvieri from Palmeiras, State of Bahia, Brazil. Analysis of the nucleotide sequence revealed that the satellite DNA obtained by digestion with EcoRI, called PcP190EcoRI, is 70% similar to the coding region of type I 5S rDNA and 66% similar to the coding region of type II 5S rDNA. Membrane hybridization and PCR amplification of the sequence showed that PcP190EcoRI is tandemly repeated. The satellite DNA as well as type I and type II 5S rDNA were localized in P. cuvieri chromosomes by fluorescent in situ hybridization. The PcP190EcoRI sequence was found in the centromeres of chromosomes 1-5 and in the pericentromeric region of chromosome 3. Type I 5S rDNA was detected in chromosome 3, coincident with the site of PcP190EcoRI. Type II 5S rDNA was located interstitially in the long arm of chromosome 5. None of these sequences co-localized with nucleolar organizer regions. Our data suggests that this satellite DNA originates from the 5S ribosomal multigene family, probably by gene duplication, nucleotide divergence and sequence dispersion in the genome.


Use of a bifurcated ePTFE graft for off-pump extracardiac Fontan completion.
A 6-year-old girl underwent off-pump extracardiac Fontan completion. Intrapericardial inferior caval vein was found to be accompanied by the hepatic vein on the left side; a 20 x 10 x 10-mm bifurcated ePTFE graft was used without external shunting. This approach is practical for the surgeons and may result in lower perioperative Fontan pressures/transpulmonary gradients as well as a shorter intubation period and hospital stay.


Ancillary service impact on outpatient scheduling.
The paper aims to provide a simulation optimization solution to improve patient scheduling that accounts for varying ancillary service time such as x-ray to minimize patient wait time. The two-step approach is to: identify patients' needs for ancillary services while scheduling appointments; and propose an algorithm to determine ancillary service time via simulation optimization. The main aim is to provide sufficient time between arrival at the clinic and the actual examination time for a patient to complete pre-visit activities without contributing significantly to patient wait time. Two case studies are included to demonstrate the approach. Triaging at the appointment-scheduling time saves an average 17 minutes for physician's first consultation in a clinic day, and a 7 percent reduction on current average patient wait time for case 1. Case 2 results in a 9 percent reduction on average patient wait time. The scheduled ancillary service time depends on the frequency and the ancillary service time, and appointment slot design. One limitation is the impact of modeling error on the account of ancillary service times and the modeling assumptions. The proposed approach provides a studying method for clinic staff to account for ancillary services prior to physicians' visits for a better patient care. Two case studies demonstrated the practicability and promising results on reducing patient waiting. This article presents a unique approach to considering the required ancillary services in outpatient scheduling system that minimizes patient wait times. The approach will strengthen the existing scheduling methods to allow the time for ancillary services.


Cross-faded: Young Adults' Language of Being Simultaneously Drunk and High.
The term "cross-faded," observed in focus groups and on a variety of websites, appears to refer to overlapping drug effects of multiple substances used at the same time, particularly alcohol and marijuana. This study explores young adult understanding of the cross-faded terminology in order to inform substance use research and intervention. Young adult participants (N=807, ages 18-23) in the screening survey for Project Transitions based in Seattle, WA were asked whether they had heard of being cross-faded, what they thought it meant, and how desirable and risky they thought it was. Cross-faded was a commonly understood term (87% had heard of it), most often described as using alcohol and marijuana simultaneously (43%) and second most as being both drunk and high (25%), specifically. Being cross-faded was seen as moderately risky and not desirable by most, although 18.2% described it as moderately or very desirable. Risk factor differences in perceptions of being cross-faded were found for sex, college status, and alcohol and marijuana use. Cross-faded is a common term for the effects of using multiple substances. As such it merits further research consideration with the aim of optimizing the effectiveness of surveys and programming.


[Development of a guideline for rehabilitation of patients with low back pain-- phase 2: analysis of data of the classification of therapeutic procedures].
Initiated by the Federal Insurance Institute for Salaried Employees (BfA, Bundesversicherungsanstalt für Angestellte), the project is aimed at developing an evidence-based guideline for rehabilitation of patients with low back pain (LBP). Guideline development will be based on a systematic review of the literature, an analysis of the treatment procedures currently employed in rehabilitation, inclusion of the patients' perspectives as well as consultation of experts' panels of clinically experienced physicians and therapists. Formulation of the guideline will then be carried out in a structured consensus building process. This article is focused on analysing the present situation with regard to the treatments received by patients insured by the BfA, using data from routine documentation according to the Classification of Therapeutic Procedures (KTL, Klassifikation Therapeutischer Leistungen). The analysis is intended to provide indications of a basic need to implement a common guideline as well as, simultaneously, to explore possible deficits in present treatment practices, hence to define priorities requiring special attention in the framework of guideline development. As a result of a systematic literature review, the KTL-defined therapeutic procedures that had emerged as relevant in the rehabilitation of LBP patients were aggregated into so-called therapeutic modules which then formed the basis of the analysis. In all, more than 46,000 KTL-data of 2438 patients with a diagnosis of "low back pain" (M54.5 ICD-10) were included. In the rehab centres investigated, rehabilitation of patients with LBP follows a multidimensional, multiprofessional therapeutic strategy. More than 90 % of all patients receive treatments from the modules "medical training therapy", "health education", "physiotherapy", and "physical therapy". Treatments provided to a majority of the patients are massage (78 %), electrotherapy (67 %) as well as psychotherapy (68 %). Women more frequently than men receive therapies belonging to the "psychological treatments" and "occupational therapies" modules; younger patients receive more of the "training therapies", and more often. When treatments are compared across centres, a large variability in the therapeutic procedures provided becomes apparent. This high degree of variability suggests that development and implementation of a common clinical practice guideline for rehabilitation of patients with chronic low back pain should receive priority attention.


Targeting TAM-secreted S100A9 effectively enhances the tumor-suppressive effect of metformin in treating lung adenocarcinoma.
Metformin's effect on tumor treatment was complex, because it significantly reduced cancer cell proliferation in vitro, but made no difference in prognosis in several clinical cohorts. Our transcriptome sequencing results revealed that tumor-associated macrophage (TAM) infiltration significantly increased in active lung adenocarcinoma (LUAD) patients with long-term metformin use. We further identified that the tumor suppressive effect of metformin was more significant in mice after the depletion of macrophages, suggesting that TAMs might play an important role in metformin's effects in LUAD. Combining 10X Genomics single-cell sequencing of tumor samples, transcriptome sequencing of metformin-treated TAMs, and the ChIP-Seq data of the Encode database, we identified and validated that metformin significantly increased the expression and secretion of S100A9 of TAMs through AMPK-CEBP/β pathway. For the downstream, S100A9 binds to RAGE receptors on the surface of LUAD cells, and then activates the NF-κB pathway to promote EMT and progression of LUAD, counteracting the inhibitory effect of metformin on LUAD cells. In cell-derived xenograft models (CDX) and patient-derived xenograft models (PDX) models, our results showed that neutralizing antibodies targeting TAM-secreted S100A9 effectively enhanced the tumor suppressive effect of metformin in treating LUAD. Our results will enable us to better comprehend the complex role of metformin in LUAD, and advance its clinical application in cancer treatment.


Adsorption of CO2 on the ω-Fe (0001) surface: insights from density functional theory.
The stabilization of a hexagonal phase known as the ω-phase in steel has recently been identified. The presence of C in steel samples is found to be helping the formation of this otherwise meta stable phase. This indicates that the probability of degradation of the surface is high in steel samples containing the ω-phase, through surface adsorption. Here we calculate the adsorption process of CO2 on the ω-Fe(0001) surface, for different sites and find that it strongly adsorbs horizontally with a bent configuration. The adsorption is characterized by significant charge transfer from the surface Fe atoms to the CO2 molecule, and structural modification of the molecule is occurring. The density of states calculations indicate that hybridization and subsequent charge transfer is probable between the d orbitals of Fe and p orbitals of CO2, resulting in strong chemisorption, that further leads to spontaneous dissociation of the molecule.


The effects of tissue proportions on blood volume change-induced variations using bio-impedance analysis: a simulation study.
Bioimpedance Analysis (BIA) along the radial artery has been widely investigated for hemodynamic monitoring. However, its applicability to different body type populations still lacks sufficient research. The Finite Element Method (FEM) was performed on three different wrist models using ANSYS HFSS, aiming to reveal the influences of different fat and muscle proportions on the sensitivity of blood volume change-induced bioimpedance change. The simulation results confirmed that the current density in each tissue mainly depended on the conductivity of tissues. The higher conductivity of the tissue, the higher current density inside said tissue. The amounts of flowing current were decided by both volume and conductivity of tissues. Moreover, increasing the fat layer thickness from 4 mm to 6 mm raised simulated impedance from 86.82 Ω to 100.39 Ω and impedance change from 0.63 Ω to 1.55 Ω. However, a higher muscle proportion occupied more injected current from the blood and resulted in lower impedance change. Therefore, for the overweight population, the placement of BIA is recommended to avoid the muscular body parts for the acquirement of better-quality pulse waves.Clinical Relevance-This establishes the bio-impedance analysis should avoid the muscular body parts for a better blood pulse wave quality for overweight populations.


Hospital IT adoption strategies associated with implementation success: implications for achieving meaningful use.
Health systems are facing significant pressure to either implement health information technology (HIT) systems that have "certified" electronic health record applications and that fulfill the federal government's definition of "meaningful use" or risk substantial financial penalties in the near future. To this end, hospitals have adopted one of three strategies, described as "best of breed," "best of suite," and "single vendor," to meet organizational and regulatory demands. The single-vendor strategy is used by the simple majority of U.S. hospitals, but is it the most effective mode for achieving full implementation? Moreover, what are the implications of adopting this strategy for achieving meaningful use? The simple answer to the first question is that the hospitals using the hybrid best of suite strategy had fully implemented HIT systems in significantly greater proportions than did hospitals employing either of the other strategies. Nonprofit and system-affiliated hospitals were more likely to have fully implemented their HIT systems. In addition, increased health maintenance organization market penetration rates were positively correlated with complete implementation rates. These results have ongoing implications for achieving meaningful use in the near term. The federal government's rewards and incentives program related to the meaningful use of HIT in hospitals has created an organizational imperative to implement such systems. For hospitals that have not begun systemwide implementation, pursuing a best of suite strategy may provide the greatest chance for achieving all or some of the meaningful use targets in the near term or at least avoiding future penalties scheduled to begin in 2015.


Role of Sigma Receptor in Cocaine-Mediated Induction of Glial Fibrillary Acidic Protein: Implications for HAND.
Cocaine abuse has been shown to accelerate the progression of human immunodeficiency virus (HIV)-1-associated neurological disorders (HANDs) partially through increasing neuroinflammatory response mediated by activated astrocytes; however, the detailed molecular mechanism of cocaine-mediated astrocyte activation is unclear. In the current study, we demonstrated increased astrogliosis in the cortical regions of brains from HIV(+) cocaine abusers compared with the HIV(+) group without cocaine abuse. We next sought to explore whether cocaine exposure could result in increased expression of glial fibrillary acidic protein (GFAP), a filament protein critical for astrocyte activation. Exposure of cocaine to astrocytes resulted in rapid translocation of sigma receptor to the plasma membrane with subsequent activation of downstream signaling pathways. Using a pharmacological approach, we provide evidence that cocaine-mediated upregulation of GFAP expression involved activation of mitogen-activated protein kinase (MAPK) signaling with subsequent downstream activation of the early growth response gene 1 (Egr-1). Egr-1 activation, in turn, caused transcriptional regulation of GFAP. Corroboration of these findings in vivo demonstrated increased expression of GFAP in the cortical region of mice treated with cocaine compared with the saline injected controls. A thorough understanding of how cocaine mediates astrogliosis could have implications for the development of therapeutic interventions aimed at HIV-infected cocaine abusers.


Low-cost production of proinsulin in tobacco and lettuce chloroplasts for injectable or oral delivery of functional insulin and C-peptide.
Current treatment for type I diabetes includes delivery of insulin via injection or pump, which is highly invasive and expensive. The production of chloroplast-derived proinsulin should reduce cost and facilitate oral delivery. Therefore, tobacco and lettuce chloroplasts were transformed with the cholera toxin B subunit fused with human proinsulin (A, B, C peptides) containing three furin cleavage sites (CTB-PFx3). Transplastomic lines were confirmed for site-specific integration of transgene and homoplasmy. Old tobacco leaves accumulated proinsulin up to 47% of total leaf protein (TLP). Old lettuce leaves accumulated proinsulin up to 53% TLP. Accumulation was so stable that up to ~40% proinsulin in TLP was observed even in senescent and dried lettuce leaves, facilitating their processing and storage in the field. Based on the yield of only monomers and dimers of proinsulin (3 mg/g leaf, a significant underestimation), with a 50% loss of protein during the purification process, one acre of tobacco could yield up to 20 million daily doses of insulin per year. Proinsulin from tobacco leaves was purified up to 98% using metal affinity chromatography without any His-tag. Furin protease cleaved insulin peptides in vitro. Oral delivery of unprocessed proinsulin bioencapsulated in plant cells or injectable delivery into mice showed reduction in blood glucose levels similar to processed commercial insulin. C-peptide should aid in long-term treatment of diabetic complications including stimulation of nerve and renal functions. Hyper-expression of functional proinsulin and exceptional stability in dehydrated leaves offer a low-cost platform for oral and injectable delivery of cleavable proinsulin.


Getting patient blood management Pillar 1 right in the Asia-Pacific: a call for action.
Preoperative anaemia is common in the Asia-Pacific. Iron deficiency anaemia (IDA) is a risk factor that can be addressed under patient blood management (PBM) Pillar 1, leading to reduced morbidity and mortality. We examined PBM implementation under four different healthcare systems, identified challenges and proposed several measures: (a) Test for anaemia once patients are scheduled for surgery. (b) Inform patients about risks of preoperative anaemia and benefits of treatment. (c) Treat IDA and replenish iron stores before surgery, using intravenous iron when oral treatment is ineffective, not tolerated or when rapid iron replenishment is needed; transfusion should not be the default management. (d) Harness support from multiple medical disciplines and relevant bodies to promote PBM implementation. (e) Demonstrate better outcomes and cost savings from reduced mortality and morbidity. Although PBM implementation may seem complex and daunting, it is feasible to start small. Implementing PBM Pillar 1, particularly in preoperative patients, is a sensible first step regardless of the healthcare setting.


Plasma Protein Pattern Correlates With Pain Intensity and Psychological Distress in Women With Chronic Widespread Pain.
Objectives: Although generalized muscle pain, tiredness, anxiety, and depression are commonly present among chronic widespread pain (CWP) patients, the molecular mechanisms behind CWP are not fully elucidated. Moreover, the lack of biomarkers often makes diagnosis and treatment problematic. In this study, we investigated the correlation between pain intensity, psychological distress, and plasma proteins among CWP patients and controls (CON). Methods: The plasma proteome of CWP (n = 15) and CON (n = 23) was analyzed using two-dimensional gel electrophoresis. Orthogonal Partial Least Square analysis (OPLS) was used to determine proteins associated with pain intensity (numeric rating scale) in CWP and psychological distress (Hospital and Depression Scale, HADS) in CWP and CON. Significant proteins were identified by MALDI-TOF and tandem MS. Results: In CWP, pain intensity was associated with plasma proteins mostly involved in metabolic and immunity processes (e.g., kininogen-1, fibrinogen gamma chain, and ceruloplasmin), and psychological distress was associated with plasma proteins related to immunity response, iron ion, and lipid metabolism (e.g., complement factor B, complement C1r subcomponent, hemopexin, and clusterin). Discussion: This study suggests that different plasma protein patterns are associated with different pain intensity and psychological distress in CWP. Proteins belonging to the coagulation cascade and immunity processes showed strong associations to each clinical outcome. Using the plasma proteome profile of CWP to study potential biomarker candidates provides a snapshot of ongoing systemic mechanisms in CWP.


Promoter selectivity of the stationary-phase forms of Escherichia coli RNA polymerase and conversion in vitro of the S1 form enzyme into a log-phase enzyme-like form.
Upon growth transition of Escherichia coli cells from exponential to stationary phase, RNA polymerase is converted into at least three different forms (S1, S2 and S3), which could be separately isolated by phosphocellulose column chromatography (Ozaki et al., 1991 (2)). Here, the promoter selectivity of these three stationary-phase enzymes was examined using an in vitro mixed transcription system and an E. coli promoter collection. These altered forms of RNA polymerase showed different recognition properties of promoters from that by the log-phase holoenzyme (L1). One of the stationary-phase RNA polymerases, S1, was found to be converted in vitro into an enzyme like the log-phase form following incubation with nucleotides or pyrophosphate. The conversion was indicated by not only the shift of elution position from a phosphocellulose column but also the change in the promoter selectivity. These results may suggest that RNA polymerase is interconvertible between different forms with different promoter selectivity by interaction with a phosphorylated compound(s).


The treatment of alcoholic withdrawal states with oxygen and nitrous oxide.
Chronic alcoholic subjects suffering withdrawal symptoms were treated with 100% oxygen or with oxygen/nitrous oxide. In most cases oxygen alone reversed withdrawal symptoms except, in a few instances, depression. The residual depressive symptoms not reversed by oxygen were completely reversed by nitrous oxide/oxygen. The pathogenesis of alcoholic withdrawal states in the light of the overactivity of the adrenergic system which leads to tissue acidosis is discussed. The importance of correct psychological management is stressed. Our therapy reduced both the period of hospitalization and the amount of sedation required. The similarity between opiate and alcohol withdrawal and treatment is mentioned.