Pathogenic role of mitogen activated protein kinases in protozoan parasites.
Protozoan parasites with complex life cycles have high mortality rates affecting billions of human lives. Available anti-parasitic drugs are inadequate due to variable efficacy, toxicity, poor patient compliance and drug-resistance. Hence, there is an urgent need for the development of safer and better chemotherapeutics. Mitogen Activated Protein Kinases (MAPKs) have drawn much attention as potential drug targets. This review summarizes unique structural and functional features of MAP kinases and their possible role in pathogenesis of obligate intracellular protozoan parasites namely, Leishmania, Trypanosoma, Plasmodium and Toxoplasma. It also provides an overview of available knowledge concerning the target proteins of parasite MAPKs and the need to understand and unravel unknown interaction network(s) of MAPK(s).


Calprotectin in rheumatoid arthritis : association with disease activity in a cross-sectional and a longitudinal cohort.
Calprotectin is potentially a more sensitive biomarker of disease activity in rheumatoid arthritis (RA) than conventional acute-phase proteins such as the erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) because it directly reflects inflammation in the synovium and synovial fluid rather than systemic inflammatory activity. The aim of this study was to evaluate relationships between serum calprotectin levels, disease activity, and response to treatment. Calprotectin was also investigated as a predictive marker of clinical response. This observational study included selected cohorts of patients with RA treated at La Paz University Hospital, Madrid, Spain. Associations between serum calprotectin levels and clinical and laboratory parameters were analyzed in a cross-sectional cohort of 60 patients with varying disease activity, and changes in calprotectin levels in response to treatment with infliximab were analyzed at baseline and after 3 and 6 months of treatment in a longitudinal cohort of 20 patients with very active disease. In the cross-sectional cohort, calprotectin levels correlated with rheumatoid factor levels (r = 0.25; p < 0.05) but not with titers of antibodies to cyclic citrullinated peptide. Significant correlations were also observed between calprotectin levels and the 28 swollen joint count (28-SJC), Disease Activity Score based on a 28-joint count (DAS28), Simplified Disease Activity Index (SDAI), ESR, and CRP levels. In the longitudinal cohort, calprotectin levels at baseline were not predictive of response to treatment but significantly decreased during treatment in responders (p < 0.0001). Calprotectin levels strongly correlate with clinical and laboratory assessments of joint inflammation and also decrease in response to treatment, indicating that calprotectin is a promising marker for assessment and monitoring of disease activity in patients with RA. Investigations are required to further evaluate its diagnostic, prognostic, and therapeutic potential.


The enhanced antibacterial effect of BNNS_Van@CS/MAO coating on Mg alloy for orthopedic applications.
The development of multifunctional Mg-based active implants with controllable degradation and antibacterial capabilities has become a hotspot in the research field of biodegradable metallic materials. To this end, a BN nanosheets (BNNS) _vancomycin (Van) @chitosan (CS) nanocomposite coating containing two antibacterial components (BNNS and Van) was prepared on Mg alloys via a micro-arc oxidation (MAO) pre-treatment combined with following electrodeposition. The related characterizations of the coating show that the composite coating has a high roughness, hydrophobicity and fair corrosion resistance. In vitro antibacterial experiments show that the BNNS_Van@CS/MAO composite coating have obvious inhibitory effect on the growth of both E. coli and S. aureus. The antibacterial effect of the BNNS_Van@CS/MAO composite coating was attributed to the synergistic effect of CS, BNNS and Van. This study provides a valuable surface modification strategy for developing multifunctional Mg-based implants with good corrosion resistance and antibacterial properties.


Genetic structure of the Atlantic Rainforest tree species Luehea divaricata (Malvaceae).
The Atlantic Rain Forest is one of the most important Brazilian biomes and a hotspot for biodiversity that is characterized by its high level of endemism, where new species are still being described. Luehea divaricata (Malvaceae) is commonly found in riparian forests areas of the Atlantic forest. Because of the importance of this species in reforestation programs, we used nine pairs of microsatellite loci to study the genetic variability of this species along its distribution area and verify if fragmentation is compromising the survival of these populations. A total of 50 alleles were obtained with an average observed and expected heterozygosity of 0.53 and 0.67, respectively. Seven of the nine populations studied showed a heterozygosity deficit. Most of the genetic diversity was found within populations; while the level of genetic differentiation was moderated (6.84) between populations. Different levels of gene flow between the populations were detected. Positive and significant values of Fis were found for seven populations. The signal test for excess of heterozygosity indicated that a recent genetic bottleneck occurred in the fragmented populations. The dendrogram constructed by the UPGMA method revealed the formation of seven clusters, which was confirmed by the Bayesian analysis for number of K clusters. The presence of several pairs of loci in linkage disequilibrium confirms that these populations experienced a loss of genetic diversity caused by genetic drift. The results showed that it is necessary to develop management strategies for the conservation of these populations of L. divaricata as the viability of the next generations are severely compromised.


In vivo measurement of [18F]fluorodeoxyglucose rate constants in rat brain by external coincidence counting.
The operational equation for the double-label deoxyglucose method described in the following paper requires the knowledge of the rate constants for transfer of fluorodeoxyglucose across the blood-brain barrier (K1* and K2*), and those for phosphorylation of fluorodeoxyglucose (K3*) and dephosphorylation of fluorodeoxyglucose-6-phosphate (k4*). These rate constants were determined in anesthetized rats by external coincidence counting. Radioactivity in parietal brain was measured for a 110 min experimental period after a bolus injection of 18F-labeled fluorodeoxyglucose. Apparent rate constants were obtained by fitting the resulting tissue radioactivity curves to the tissue radioactivity function of the deoxyglucose model modified to take into account the dephosphorylation of fluorodeoxyglucose-6-phosphate. The apparent fluorodeoxyglucose rate constants in rat brain are K1* = 0.195 ml g-1 min-1, k2* = 0.379 min-1, k3* = 0.088 min-1, and k4* = 0.009 min-1.


Nomogram to predict the risk of biochemical recurrence and structural recurrence in patients with stage cN1 papillary thyroid carcinoma.
Although papillary thyroid carcinoma (PTC) is thought to be the least aggressive thyroid cancer, it has a significant recurrence rate. Therefore, we aimed to develop a nomogram to estimate the probability of biochemical recurrence (BIR) and structural recurrence (STR) in patients with stage cN1 PTC. We studied the relationship between the characteristics of patients with stage N1a PTC and the risk of recurrence by analysing the data of 617 inpatients (training cohort) and 102 outpatients (validation cohort) in our hospital. We used the least absolute shrinkage and selection operator regression model to identify prognostic indicators to construct nomograms to predict the risk of BIR and STR. There were 94 (15.24%) BIR cases in the training cohort and 36 (35.29%) in the validation cohort. There were 31 (5.02%) STR cases in the training cohort and 23 (22.55%) cases in the validation cohort. The variables included in the BIR nomogram were sex, age at diagnosis, tumour size, extrathyroidal infiltration, and lymph node ratio (LNR). While the variables included in the STR nomogram were tumour size, extrathyroidal infiltration, BRAF state, metastatic lymph nodes, and LNR. Both the prediction models demonstrated good discrimination ability. The results showed the calibration curve of the nomogram was near the optimum diagonal line, and the decision curve analysis showed a noticeably better benefit. The LNR may be a valid prognostic indicator for patients with stage cN1 PTC. The nomograms could help clinicians identify high-risk patients and choose the best postsurgical therapy and monitoring.


Is There An Advantage of Using Dingkun Pill () alone or in Combination with Diane-35 for Management of Polycystic Ovary Syndrome? A Randomized Controlled Trial.
To evaluate the effects of Chinese medicine Dingkun Pill () alone or in combination with Diane-35 on patients with polycystic ovary syndrome (PCOS). This is a prospective randomized controlled trial conducted at Peking Union Medical College Hospital Beijing, China, from December 2016 to September 2017. Totally 117 PCOS patients were randomly assigned to the Dingkun Pill group (38 cases), Diane-35 group (40 cases), or combined group (39 cases). Patients in the Dingkun Pill group or Diane-35 group took daily 7 g of oral Dingkun Pill or 1 tablet of oral Diane-35, respectively, for 21 consecutive days followed by 7 drug-free days. And the combined group received a combination of Dingkun Pill and Diane-35. The treatment course was 3 months. Fasting plasma glucose and insulin, total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triglycerides (TG), free fatty acids (FFA) and sex hormones were analyzed, quantitative insulin sensitivity check index (QUICKI) was calculated, and menstruation and acne scores were recorded at baseline and after 3-month treatment. Compared with before treatment, QUICKI decreased significantly in the Dingkun Pill and combined groups after 3-month treatment (P<0.05); TC, LDL-C and FFA decreased significantly in the Dingkun Pill group (P<0.01), LDL-C also decreased obviously in the Diane-35 group (P<0.01), while TC increased significantly in the combined group (P<0.01), TG increased significantly in all groups (P<0.01); total testosterone (TT) and menstruation regularity was improved significantly in the Diane-35 and combined groups (P<0.01); acne scores were improved in all groups (P<0.01). After treatment, TC and FFA in the Dingkun Pill group were significantly lower than the Diane-35 group (P<0.05 or P<0.01); TT was lower and regular menstruation rate was higher in the Diane-35 and combined groups than the Dingkun Pill group (P<0.01), and no differences were observed between Diane-35 group and combined group (P>0.05). Dingkun Pill showed better effects than Diane-35 in improving insulin sensitivity, lowering TC and FFA. Diane-35 was more efficient in regulating menstruation and lowering androgen than Dingkun Pill. Combination of Dingkun Pill and Diane-35 may be a better choice to regulate menstruation, lower androgens while improve glucose metabolism in PCOS patients. (Registered on ClinicalTrials.gov, registration No. NCT03264638).


Construction and immunological evaluation of truncated hepatitis B core particles carrying HBsAg amino acids 119-152 in the major immunodominant region (MIR).
Hepatitis B capsid protein expressed in Escherichia coli can reassemble into icosahedral particles, which could strongly enhance the immunogenicity of foreign epitopes, especially those inserted into its major immunodominant region. Herein, we inserted the entire 'α' antigenic determinant amino acids (aa) 119-152 of HBsAg into the truncated HBc (aa 1-144), between Asp(78) and Pro(79). Prokaryotic expression showed that the mosaic HBc was mainly in the form of inclusion bodies. After denaturation with urea, it was dialyzed progressively for protein renaturation. We observed that before and after renaturation, mosaic HBc was antigenic as determined by HBsAg ELISA and a lot of viruslike particles were observed after renaturation. Thus, we further purified the mosaic viruslike particles by (NH4)2SO4 precipitation, DEAE chromatography, and Sepharose 4FF chromatography. Negative staining electron microscopy demonstrated the morphology of the viruslike particles. Immunization of Balb/c mice with mosaic particles induced the production of anti-HBs antibody and Th1 cell immune response supported by ELISPOT and CD4/CD8 proportions assay. In conclusion, we constructed mosaic hepatitis core particles displaying the entire 'α' antigenic determinant on the surface and laid a foundation for researching therapeutic hepatits B vaccines.


Combining CD34+ stem cell selection with prophylactic pathogen and leukemia directed T-cell immunotherapy to simultaneously reduce graft versus host disease, infection, and leukemia recurrence after allogeneic stem cell transplant.
We designed a trial to simultaneously address the problems of graft versus host disease (GVHD), infection, and recurrence of malignancy after allogeneic stem cell transplantation. CD34+ stem cell isolation was used to minimize the development of acute and chronic GVHD. Two prophylactic infusions, one combining donor-derived cytomegalovirus, Epstein-Barr virus, and Aspergillus fumigatus specific T-cells and the other comprising donor-derived CD19 directed chimeric antigen receptor (CAR) bearing T-cells, were given 21-28 days after transplant. Two patients were transplanted for acute lymphoblastic leukemia from HLA identical siblings using standard doses of cyclophosphamide and total body irradiation without antilymphocyte globulin. Patients received no post-transplant immune suppression and were given no pre-CAR T-cell lymphodepletion. Neutrophil and platelet engraftment was prompt. Following adoptive T-cell infusions, there was rapid appearance of antigen-experienced CD8+ and to a lesser extent CD4+ T-cells. Tetramer-positive T-cells targeting CMV and EBV appeared rapidly after T-cell infusion and persisted for at least 1 year. CAR T-cell expansion occurred and persisted for up to 3 months. T-cell receptor tracking confirmed the presence of product-derived T-cell clones in blood targeting all three pathogens. Both patients are alive over 3 years post-transplant without evidence of GVHD or disease recurrence. Combining robust donor T-cell depletion with directed T-cell adoptive immunotherapy targeting infectious and malignant antigens permits independent modulation of GVHD, infection, and disease recurrence. The combination may separate GVHD from the graft versus tumor effect, accelerate immune reconstitution, and improve transplant tolerability.


Cephalic vein cutdown at the wrist: comparison to the standard saphenous vein ankle cutdown.
We first developed a technique for cutdown of the cephalic vein at the wrist. We then conducted a prospective cross-over cadaver study comparing the ability of medical students to perform this technique with that of the standard saphenous vein cutdown at the ankle. All students had a previous course in anatomy but had never performed a cutdown. Before testing, the students were given written material and a ten-minute lecture describing both approaches. Seventeen students performed 34 cutdowns; nine students attempted the cephalic cutdown followed by the saphenous cutdown; the remaining eight used the reverse order. The mean time (+/- SD) to isolation of the cephalic vein was 85 +/- 70 seconds; for the saphenous vein, mean time was 70 +/- 89 seconds (P = NS). There was one failure (inability to isolate the vein within five minutes) in 17 attempts at the cephalic vein and two failures in 17 attempts at the saphenous (P = NS). There were no complications (nerve, artery, or tendon injury) with either technique. The mean external vein diameter (+/- SD) of the cephalic vein and the saphenous vein were 3.2 +/- 1.0 mm and 3.6 +/- 0.7 mm, respectively (P = NS). We conclude that relatively inexperienced providers can learn to perform the cephalic vein cutdown at the wrist on fresh cadavers with similar speed and success as that for the saphenous vein cutdown at the ankle. Cutdown at this site may provide a useful alternative to the saphenous cutdown in certain clinical situations.


Characterization of a T-DNA promoter trap line of Arabidopsis thaliana uncovers a cryptic bi-directional promoter.
Investigation of the transgenic Arabidopsis promoter trap line GFP-868 that showed GFP expression only in anthers revealed the T-DNA insertion at 461bp upstream to the hypothetical gene At4g10596 with the GFP reporter gene in head-to-head orientation to the At4g10596 gene. The expression of the At4g10596 gene in wild type and in GFP-868 plant homozygous for T-DNA insertion was comparable and found in all tissues tested, while the GFP expression was restricted to anthers of the GFP-868 plants suggesting that the 461bp fragment separating the two genes in the GFP-868 line is functioning as bi-directional promoter. This 461bp fragment was cloned upstream to the GUS gene in two orientations to test for bi-directional promoter activity. Transgenic Arabidopsis plants carrying either of these constructs showed GUS activity in anthers indicating that this fragment behaves as bi-directional promoter specific to anthers. These results were also supported by the presence of cis-acting motifs such as TATA box and POLLEN1LELAT52 (AGAAA) within the 461bp sequence in both orientations. However, transcripts corresponding to the upstream sequences beyond -461 nucleotides were not detected in the wild type suggesting that this 461bp fragment is a cryptic promoter. The significance of the promoter trap approach and the usefulness of this type of promoter are discussed.


Epicardial fat volume is related to the degree of cardiac allograft vasculopathy.
Increasing evidence suggests a role for epicardial fat in the development of coronary artery disease in the general population. Heart transplantation patients are at increased risk of developing a specific form of coronary artery disease, cardiac allograft vasculopathy (CAV), which has far-reaching consequences in terms of morbidity and mortality. Until now, the role of epicardial fat volume (EFV) in the development of CAV remains unknown. Hence, we investigated the relationship between EFV and CAV as well as the influence of donor/recipient sex on EFV. Adult heart transplant patients who underwent coronary computed tomography angiography (CCTA) for CAV screening who were four or more years post-HT were included. Using the CT examinations, we quantified the EFV and the degree of CAV. Ordinal and linear regression models were used to assess the association of EFV with CAV. In total, 149 (median age 44.5 years, 36% women) patients were included. The median time between HT and the CT scan was 11.0 (7.3-16.1) years. CAV grade 0, 1, 2 and 3 were seen in 85 (57%), 32 (22%), 14 (9%), and 18 (12%) patients, respectively. The median EFV was 208.4 (128.9-276.0) mL. Larger EFV were related to higher degrees of CAV (median of 164.7 to 290.6 mL for CAV grade 0 and 3, respectively, OR 5.23 (2.47-11.06), p < 0.001). Male recipients had significantly more EFV than female recipients irrespective of the donor sex (232.7 mL vs. 147.2 mL respectively, p < 0.001). Determinants for EFV were recipient sex, number of rejections, donor age, time between HT and CT scan, recipient BMI, and diabetes mellitus. EFV was associated with higher degrees of CAV. The recipient sex influenced the EFV more than the donor sex. • Patients after heart transplantation have a high amount of epicardial fat while larger amounts of epicardial fat are related to higher grades of cardiac allograft vasculopathy. • Determinants of higher epicardial fat volume included recipient sex, number of rejections, donor age, time between HT and CT scan, recipient BMI, and diabetes mellitus. • Longitudinal studies are needed to further disentangle the role of epicardial fat in the development and progression of cardiac allograft vasculopathy. Demonstration of four patients (from CAV grade 0 to CAV grade 3) in whom epicardial fat volume was determined. In red, the voxels identified as epicardial fat.


Oxidative stability, oxidation pattern and α-tocopherol response of docosahexaenoic acid (DHA, 22:6n-3)-containing triacylglycerols and ethyl esters.
DHA is most often used in supplements either in its triacylglycerol or ethyl ester form. Currently, there is only little published data on the differences in the oxidative stability and α-tocopherol response between the two lipid structures, as well as on the oxidation patterns of pure DHA. This study investigated the oxidative stability, α-tocopherol response and oxidation pattern of DHA incorporated in triacylglycerols and as ethyl esters with an untargeted approach after oxidation at 50 °C in the dark. Liquid and gas chromatographic methods with mass spectrometric detection and nuclear magnetic resonance spectroscopy were applied. DHA was more stable in triacylglycerols than as ethyl esters without α-tocopherol addition. With α-tocopherol added the opposite was observed. The oxidation products formed during triacylglycerol and ethyl ester oil oxidation were mostly similar, but also some structure-related differences were detected in both volatile and non-volatile oxidation products.


ETS-1-activated LINC01016 over-expression promotes tumor progression via suppression of RFFL-mediated DHX9 ubiquitination degradation in breast cancers.
Long non-coding RNAs (lncRNAs) are key regulators during the development of breast cancer (BC) and thus may be viable treatment targets. In this study, we found that the expression of the long intergenic non-coding RNA 01016 (LINC01016) was significantly higher in BC tissue samples with positive lymph node metastasis. LINC01016, which is activated by the transcription factor ETS-1, contributes to the overt promotion of cell proliferation activity, enhanced cell migratory ability, S phase cell cycle arrest, and decreased apoptosis rate. By RNA pull-down assays and mass spectrometry analyses, we determined that LINC01016 competitively bound and stabilized DHX9 protein by preventing the E3 ubiquitin ligase RFFL from binding to DHX9, thereby inhibiting DHX9 proteasomal degradation. This ultimately led to an increase in intracellular DHX9 expression and activated PI3K/AKT signaling, with p-AKT, Bcl-2, and MMP-9 involvement. This is the first study to reveal that the LINC01016/DHX9/PI3K/AKT axis plays a critical role in the progression of BC, and thus, LINC01016 may serve as a potential therapeutic target for patients with BC.


Malnutrition and gastroenteritis in The Gambia.
There is a strong association between failure to thrive and diarrhoeal disease in young Gambian children. The high prevalence of diarrhoeal disease seen is not due to frequent attacks of "acute infantile diarrhoea" of viral orgin. The picture of protracted diarrhoea is almost certainly due to colonization of the upper bowel and the high prevalence demonstrated to be at least partly due to the environment to which the children are exposed, particularly in terms of food and water hygiene. Until this cycle of upper bowel colonization and protracted diarrhoea is broken, a diet-based nutrition programme cannot be expected to function effectively.


Exergames vs. traditional exercise: investigating the influencing mechanism of platform effect on subthreshold depression among older adults.
This study aimed to examine the influencing mechanism of exercise platform effect on subthreshold depression among older adults by comparing exergames and traditional exercise. One hundred and two Singaporean older adults were assigned to either playing Wii exergames or performing traditional exercise programs once a week, for six weeks. Results confirmed a direct negative platform effect on subthreshold depression and further supported the mediation role of positive emotions in the platform effect. It implied that exergames led to higher positive emotions than traditional exercise, which further reduced the subthreshold depression among older adults. Self-efficacy was not supported to be a significant mediator in the relations between exercise platform and subthreshold depression. A better understanding of the mechanisms behind the antidepressant effects of exercise platform would not only provide additional insight into a possible causal association, but also inspire the future use of exergames in the treatment of subthreshold depression.


Biodiversity: What makes a city bird?
The impacts of urbanisation on biodiversity varies greatly across species. A new study shows how the intrinsic species properties underlying urban tolerance vary globally according to environmental context. This has important implications for conserving biodiversity in a rapidly urbanising world.


Prenatal alcohol exposure and attention, learning and intellectual ability at 14 years: a prospective longitudinal study.
A range of adverse birth outcomes is associated with heavy prenatal alcohol exposure. To examine the effects of moderate levels of alcohol consumption during pregnancy on children's intellectual ability, learning and attention at 14 years of age. The Mater-University of Queensland Study of Pregnancy involves a prospective birth cohort of 7223 singletons whose mothers were enrolled at the first antenatal visit. At 14 years, 5139 mothers and adolescents completed attentional and learning questionnaires, and 3731 adolescents completed psychometric assessments. For adolescents, the Wide Range Achievement Test--Revised (WRAT-R) and Raven's Standard Progressive Matrices Test (Raven's) were administered. Mothers completed the Child Behaviour Checklist (CBCL) and adolescents completed the Youth Self Report (YSR). Learning was assessed by a series of questions in the mother and adolescent questionnaires. Maternal measures included the quantity and frequency of alcohol consumption, and the extent of binge drinking. For consumption of <1 glass/day in early or late pregnancy, there was no association with any attention, learning or cognitive outcomes. The strongest estimates of effect were found among those consuming > or =1 glasses/day. Exposure in late pregnancy was associated with increased prevalence of overall learning difficulty in the unadjusted, although not the adjusted analysis. Binge drinking was associated with a higher prevalence of Raven's score <85 (1 standard deviation). Although a number of study limitations need to be considered, the results suggest that consumption at the level of <1 drink/day does not lead to adverse outcomes in relation to attention, learning and cognitive abilities, as measured in the current research.


A direct comparison of spine rotational stiffness and dynamic spine stability during repetitive lifting tasks.
Stability of the spinal column is critical to bear loads, allow movement, and at the same time avoid injury and pain. However, there has been a debate in recent years as to how best to define and quantify spine stability, with the outcome being that different methods are used without a clear understanding of how they relate to one another. Therefore, the goal of the present study was to directly compare lumbar spine rotational stiffness, calculated with an EMG-driven biomechanical model, to local dynamic spine stability calculated using Lyapunov analyses of kinematic data, during a series of continuous dynamic lifting challenges. Twelve healthy male subjects performed 30 repetitive lifts under three varying load and three varying rate conditions. With an increase in the load lifted (constant rate) there was a significant increase in mean, maximum, and minimum spine rotational stiffness (p<0.001) and a significant increase in local dynamic stability (p<0.05); both stability measures were moderately to strongly related to one another (r=-0.55 to -0.71). With an increase in lifting rate (constant load), there was also a significant increase in mean and maximum spine rotational stiffness (p<0.01); however, there was a non-significant decrease in the minimum rotational stiffness and a non-significant decrease in local dynamic stability (p>0.05). Weak linear relationships were found for the varying rate conditions (r=-0.02 to -0.27). The results suggest that spine rotational stiffness and local dynamic stability are closely related to one another, as they provided similar information when movement rate was controlled. However, based on the results from the changing lifting rate conditions, it is evident that both models provide unique information and that future research is required to completely understand the relationship between the two models. Using both techniques concurrently may provide the best information regarding the true effects of (in) stability under different loading and movement scenarios, and in comparing healthy and clinical populations.


Lack of association between cytomegalovirus infection, HLA matching and the vanishing bile duct syndrome after liver transplantation.
In this study we evaluated the association between cytomegalovirus infection alone or in relation to human leukocyte antigen matching and the development of vanishing bile duct syndrome, a form of chronic hepatic allograft rejection. A total of 81 consecutive liver transplant recipients were studied. Cytomegalovirus infection developed in 46 recipients (57%), and vanishing bile duct syndrome occurred in 9 recipients (11%). Cytomegalovirus infection developed in only five of the nine patients with vanishing bile duct syndrome. Univariate analysis of pretransplant recipient/donor cytomegalovirus serological tests and human leukocyte antigen typing showed they were not significant risk factors for the development of vanishing bile duct syndrome. Time-dependent analysis of cytomegalovirus infection after transplantation as a risk factor for vanishing bile duct syndrome, in a multivariate analysis with human leukocyte antigen match, showed no statistical significance. In our study, no association was found between cytomegalovirus infection alone or in relation to class I or II human leukocyte antigen match and the subsequent development of vanishing bile duct syndrome.


Framework to manage railway noise exposure in Brazil based on field measurements and strategic noise mapping at the local level.
Rail freight transport has grown worldwide and in Brazil as well, which increases people's exposure to railway noise. A promising tool to manage it is Strategic Noise Mapping (SNM), which has advanced around the world favoring a common and more accurate calculation method that requires more accurate measurements. This paper presents a framework to manage railway noise exposure in Brazil based on a case study carried out in the city with the longest stretches of railway tracks in urban areas. Background noise due to road traffic and train pass-by noise levels were measured for knowledge of noise sources and SNM calibration. Background noise predicted by the CNOSSOS-EU (Road) method reached an accuracy within ±2 dB(A) and was overestimated by the NMPB-96 method. The combination with railway noise using the SRM II and ISO 9613 calculation methods maintained the aforementioned accuracy, while the current best fit CNOSSOS-EU (Railway and Industry) configuration for the study area overestimated it. Although the study area is a quiet suburb, a quarter of its population is exposed to road traffic noise levels above those recommended by WHO, and more than 40% are affected by rail noise that is 5 dB(A) or higher than the background noise. The elimination of level crossings (LC) and the need to sound the train horn is more cost-effective than noise barriers. Therefore, the strategy to manage exposure to rail noise in Brazil should involve altering residential zoning of non-built-up areas next to the railways to repurpose them for other land uses and/or establish acoustic performance criteria for new buildings; federal government investing in reducing the number of LC or railway variants; and railway operators investing in the creation of an emission database for noise control at the source.


Neurochemical and behavioural investigations of the NMDA receptor-associated glycine site in the rat striatum: functional implications for treatment of parkinsonian symptoms.
The glutamatergic cortico-striatal and subthalamo-entopeduncular pathways are both overactive in parkinsonism. Previous behavioural investigations have shown that intra-entopeduncular injection of either NMDA-site or glycine-site antagonists results in alleviation of parkinsonian symptoms, although injection of the former is associated with the appearance of anaesthetic-like side effects. These behavioural differences may be mediated by action on different NMDA receptor subtypes. Recent neurochemical and molecular pharmacological studies have indicated the existence of NMDA receptor subtypes which display differential modulation by glycine. In the present study, three potential modes of NMDA antagonism were differentiated in vitro by effects on [3H]-glycine binding to striatal sections. Specific [3H]-glycine binding was totally displaced by the glycine partial agonist (R)-HA-966; the NMDA-site antagonist D-CPP had no effect; and the NMDA-site antagonist D-AP5 displaced [3H]-glycine binding in a subpopulation of glycine sites. The anti-parkinsonian effects of (R)-HA-966, D-CPP and D-AP5 were assessed by intra-striatal injection in reserpine-treated rats and 6-OHDA-lesioned rats. Injection of (R)-HA-966 and D-CPP resulted in alleviation of parkinsonian akinesia, although the latter elicited anaesthetic-like side effects; D-AP5 was ineffective as an anti-parkinsonian agent. (R)-HA-966 was also effective as an anti-parkinsonian agent when administered systemically in the reserpine-treated rat. These data suggest that different classes of NMDA antagonist mediate different behavioural responses within the parkinsonian striatum. The behavioural response produced may depend on the exact nature of the conformational change induced by the antagonist and the location of the subtype most sensitive to that class of compound. Selection of a specific mode of NMDA receptor antagonism or targeting of striatal NMDA receptor subtypes may form the basis of a novel therapeutic approach to Parkinson's disease.


17β-estradiol ameliorates age-associated loss of fibroblast function by attenuating IFN-γ/STAT1-dependent miR-7 upregulation.
Age-related defects in fibroblast differentiation and functionality were previously shown to be associated with impaired hyaluronan (HA) synthase 2 (HAS2) and epidermal growth factor receptor (EGFR) function, as a result of upregulated microRNA-7 (miR-7) expression. In aging fibroblasts, inhibiting miR-7 prevented the dysregulation of the HA-mediated CD44/EGFR signaling pathway. Here, we investigated transcriptional upregulation of miR-7 and implicated the age-associated over-activation of JAK/STAT1 as a primary candidate. STAT1 binding sites were identified on the putative miR-7 promoter and stimulation of fibroblasts with the inflammatory cytokine, interferon-γ (IFN-γ), significantly increased miR-7 transcriptional activity and resulted in upregulated miR-7 and loss of EGFR. Additionally, we demonstrated a role for the anti-inflammatory steroid, 17β-estradiol (E2), in the attenuation of miR-7 expression. E2 stimulation promoted estrogen receptor (ER) interactions with the miR-7 putative promoter and suppressed miR-7 expression. E2 also attenuated STAT1 expression and activity. Furthermore, treatments with E2 restored fibroblast functionality, including proliferation, migration and differentiation, key events in effective wound healing. In light of our findings, we propose that the regulation of miR-7 by pro- and anti-inflammatory mediators plays a wider role than previously thought. The modulation of fibroblast functions and ultimately wound healing by miR-7 activators or inhibitors could provide realistic targets for the restoration of chronic wound healing capabilities in the elderly.


Pharmacological approaches to counter the toxicity of Dopa.
Dopa and related catecholamines and their degradation products have been demonstrated to have neurotoxic potential in a number of cellular and in vivo experiments. Several mechanisms have been hypothesized to be involved including generation of prooxidant products that subsequently oxidize membrane lipids and exposed macromolecules. We have utilized a neuronal culture of cerebellar granule cells to study the toxicity of Dopa and the ability of various neuroprotective and antiparkinsonian compounds to offer protection therefrom. This model is apparently based on the ability of Dopa to non-enzymatically induce an oxidative injury to the neuronal cultures. Evidence for this arises from the equal neurotoxic potency of L- and D-Dopa in these cells and the ability of catalase, superoxide dismutase and glutathione to protect the neurons from this toxicity. Further, we found that the neuroprotective antioxidant, PNU-101033 is more effective and potent than vitamin E and deprenyl in this regard. Similarly the D2/D3 agonist, pramipexole is also capable of blocking Dopa toxicity in this model and this effect is independent of dopamine receptor affinity as both enantiomers are equally potent in this assay but disparate in receptor affinity. Also the protection by pramipexole is accompanied by the preservation of reduced glutathione. Thus, this activity seems to be a function of the oxidation potential of pramipexole and it's consequent antioxidant property. Potent antioxidants are effective blockers of Dopa toxicity. If the mechanisms involved in this toxicity have relevance to the progression of Parkinson's pathology in Dopa treated (or untreated) patients, these compounds have the potential to alter the course of the illness.


Development of microsatellite markers for Japanese scallop (Mizuhopecten yessoensis) and their application to a population genetic study.
The Japanese scallop (Mizuhopecten yessoensis) is one of the main fishery products in Japan, but with the expansion of culture operations of the Japanese scallop, various problems have been encountered including high mortality, poor growth, poor seed production, and so on. Moreover, there is concern that many years of cultivation may have affected the genetic structure of the scallop population. To approach these problems and concerns, we developed microsatellite markers as a molecular tool for population genetic studies. By using 4 microsatellite markers as well as a mitochondrial marker, we investigated the genetic structure of samples from the islands of Hokkaido (14 populations) and Honshu (Tohoku, 3 populations) in Japan, and south Primorye (4 populations) in Russia. All the populations sampled had high genetic diversity (average expected heterozygosity, 0.7011 to 0.7622; haplotype diversity, 0.6090 to 0.8848), and almost all showed a tendency of homozygote excess, which was significant in 2 populations. Hierarchical analysis of molecular variance tests based on the microsatellite and mitochondrial markers indicated that the 3 geographic regions were genetically divergent from one another, with little evidence of divergence within regions. Homogeneity in allele frequency distributions between natural and cultured scallops and allele frequency stability over a period of 2 decades indicated that the culturing operations have probably not had a substantial effect on the genetic structure of the populations.


Association of Diaphragm Thickness and Respiratory Muscle Strength With Indices of Sarcopenia.
To evaluate the relationship between respiratory muscle strength, diaphragm thickness (DT), and indices of sarcopenia. This study included 45 healthy elderly volunteers (21 male and 24 female) aged 65 years or older. Sarcopenia indices, including hand grip strength (HGS) and body mass index-adjusted appendicular skeletal muscle (ASM/BMI), were measured using a hand grip dynamometer and bioimpedance analysis, respectively. Calf circumference (CC) and gait speed were also measured. Maximal inspiratory pressure (MIP) and maximal expiratory pressure (MEP) were obtained using a spirometer, as a measure of respiratory muscle strength. DT was evaluated through ultrasonography. The association between indices of sarcopenia, respiratory muscle strength, and DT was evaluated using Spearman's rank correlation test, and univariate and multiple regression analysis. ASM/BMI (r=0.609, p<0.01), CC (r=0.499, p<0.01), HGS (r=0.759, p<0.01), and gait speed (r=0.319, p<0.05) were significantly correlated with DT. In the univariate linear regression analysis, MIP was significantly associated with age (p=0.003), DT (p<0.001), HGS (p=0.002), CC (p=0.013), and gait speed (p=0.026). MEP was significantly associated with sex (p=0.001), BMI (p=0.033), ASM/BMI (p=0.003), DT (p<0.001), HGS (p<0.001), CC (p=0.001) and gait speed (p=0.004). In the multiple linear regression analysis, age (p=0.001), DT (p<0.001), and ASM/BMI (p=0.008) showed significant association with MIP. DT (p<0.001) and gait speed (p=0.050) were associated with MEP. Our findings suggest that respiratory muscle strength is associated with DT and indices of sarcopenia. Further prospective studies with larger sample sizes are needed to confirm these findings.


Hepatoprotective effect of apolipoprotein A4 against carbon tetrachloride induced acute liver injury through mediating hepatic antioxidant and inflammation response in mice.
Apolipoprotein A4 (ApoA4) regulates lipid and glucose metabolism and exerts anti-inflammatory effects in atherogenesis and colitis. The present study explored the presumed protective role of ApoA4 in carbon tetrachloride (CCl4)-induced acute liver injury (ALI) in mice. The ALI model in wild type (WT), ApoA4 knock-out (ApoA4-KO) and ApoA4 transgenic (ApoA4-TG) mice was induced by a single intraperitoneal administration of CCl4. Liver and blood were harvested from mice to assess liver functions, immunohistological changes, immune cell populations and cytokine profiles. ApoA4 deficiency aggravated, and ApoA4 overexpression alleviated CCl4-inflicted liver damage by controlling levels of anti-oxidant enzymes. ApoA4 deletion increased the recruitment of monocytes/macrophages into the injured liver and upregulated the plasma levels of IL-6, TNF-α and MCP-1, but lower IL-10 and IFN-γ. ApoA4 over-expression rescued this effect and resulted in lower percentages of monocytes/macrophages and dendritic cells, the ratio of blood pro-inflammatory to anti-inflammatory monocytes and reduced plasma concentrations of IL-6, but enhanced IL-10 and IFN-γ. We propose ApoA4 as a potential new therapeutic target for the management of liver damage.


Differential identification of Mycobacterium tuberculosis complex and nontuberculous mycobacteria by duplex PCR assay using the RNA polymerase gene (rpoB).
A novel duplex PCR method that can amplify the 235- and 136-bp rpoB DNAs of Mycobacterium tuberculosis complex and nontuberculous mycobacteria (NTM), respectively, with two different sets of primers was used to differentially identify 44 reference strains and 379 clinical isolates of mycobacteria in a single-step assay. Showing 100% sensitivity and specificity, the duplex PCR method could clearly differentiate M. tuberculosis complex and NTM strains. In addition, restriction fragment length polymorphism analysis and direct sequencing of the amplicon of NTM could be used to supplement species identification.


[The use of cryomassage in the rehabilitation of IHD patients].
The authors have developed a technique of abdominal cryomassage and tried it in patients with coronary heart disease (CHD). The massage improved the disease course, diastolic function, being most effective in hypertrophic left ventricular myocardial affection; produced lipotropic action important for early secondary prevention. Thus, abdominal cryomassage as a nonpharmaceutical method can serve an efficient adjuvant modality in combined treatment and rehabilitation of CHD patients.


Monitoring of filter patency during carotid artery stenting using near-infrared spectroscopy with high time-resolution.
We aimed to evaluate the usefulness of a newly developed, near-infrared spectroscopy (NIRS) device for monitoring hemodynamic changes during carotid artery stenting (CAS), as a means to detect filter obstruction due to distal embolism. We evaluated 16 patients with internal carotid artery (ICA) stenosis during the CAS procedure, using a NIRS system that can monitor not only changes in oxygenation of hemoglobin (Hb), but also the fluctuation of oxyhemoglobin (oxy-Hb) synchronized with heartbeat. The NIRS system detected a marked decrease of oxy-Hb and an increase of deoxyhemoglobin (deoxy-Hb) during ICA occlusion in patients without anterior cross circulation (ACC). Patients with ACC showed much smaller changes. The analysis of oxy-Hb fluctuation made it possible to detect occurrence of no-flow in the absence of Hb concentration changes. The amplitude of oxy-Hb fluctuation in the no/slow-flow group was significantly smaller than that in the normal-flow group. Our results indicate that the present high time-resolution NIRS device, which can measure oxy-Hb fluctuation, is superior to conventional NIRS for detecting filter obstruction.


Hemodynamic parameters following pelvic exenteration.
Hemodynamic parameters were prospectively studied in 31 patients who underwent pelvic exenteration. With the use of a thermistor-tipped pulmonary artery catheter, hemodynamic parameters were calculated during the intraoperative and acute (less than 48 hours) postoperative interval. The mean operative time was 5.5 +/- 0.8 hours, and volume replacement (mean, 21.6 ml/kg/hr) consisted of crystalloid, colloid, and blood. Postoperative urine production (mean, 1.9 ml/kg/hr) was maintained with crystalloid (mean, 2.5 ml/kg/hr), colloid (0.2 ml/kg/hr), and blood (0.4 ml/kg/hr). Despite individual variations, the important parameters of cardiovascular function were maintained in the physiologic range. No patient developed cardiovascular or respiratory failure. We believe that the lack of perioperative morbidity and mortality was related, in substantial part, to this type of cardiovascular monitoring, which allows for the prompt diagnosis of potential problems and enables the physician to make appropriate interventions to correct these problems.


Body Mass Index and Outcomes in Breast Cancer Treated With Breast Conservation.
Obesity is increasing in incidence in New Zealand. Maintaining locoregional control after breast cancer surgery is important, as this can improve overall survival. Although obesity has been associated with poorer survival in breast cancer, the correlation with locoregional control is less well evaluated and was the subject of this evaluation. We used the New Zealand breast cancer registry to identify patients treated with breast conservation with body mass index (BMI) recorded. We retrieved patient demographic, tumor characteristic, and treatment-related information. We assessed local and locoregional control outcomes in addition to breast cancer and overall survival outcomes. Patients (2513) were identified. The median follow-up was 5.25 years. Of these patients, 38% were obese with a BMI of ≥30 kg/m2, 32% were overweight with a BMI of 25.0 to 29.9 kg/m2, and 29% were either normal or underweight with a BMI of ≤24.9 kg/m2. Most patients were postmenopausal (65%). The median tumor size was 16.0 mm (0.2-80 mm) and the majority were grade 2 or 3 tumors (45% and 33%). The majority (66%) were pathologically node negative and lymphovascular invasion was seen in 24%. There were 81% with estrogen receptor positivity and 358 (14.2%) were Her-2 positive. Local relapse free survival at 5 years was 91.5% (95% confidence interval [CI], 90.3-92.7) and at 10 years was 78.5% (95% CI, 75.9-81.1). Local relapse free rate at 5 years was 96.8% (95% CI, 96.0-97.6) and 10 years was 93.9% (95% CI, 92.5-95.3). We did not find BMI to be associated with local or locoregional relapse, disease free, breast cancer specific, or overall survival. Obesity was not associated with inferior locoregional control or survival outcomes. This supports the practice of continuing to offer breast conserving treatment to women regardless of BMI.


Biological depletion of neutrophils attenuates pro-inflammatory markers and the development of the psoriatic phenotype in a murine model of psoriasis.
Although neutrophils are considered a histologic hallmark of psoriasis, their pathophysiologic role in psoriasis remains unclear. We characterized the effects of neutrophil depletion via injection of monoclonal antibody 1A8 on the development of imiquimod (IMQ)-induced psoriatic lesions in a murine model. Lesions were followed with photographs and histologic analysis, revealing reduced psoriasiform scale and epidermal hyperplasia in neutrophil-depleted. ELISA and flow cytometry were used to determine relative levels of cytokines and immune cells. Compared to controls, IMQ-treated neutropenic mice had significantly lower levels of macrophages in tissue samples (P < .05) and displayed significantly lower numbers of CD4+ T-cells (P < .05). Neutropenic animals exhibited lower levels of TNF-α, IFN-γ, and IL-1β than controls (P < .05). These results show that neutropenia reduces the development of psoriasiform skin lesions and substantially decreases infiltration of pro-inflammatory cytokines and immune cells to IMQ-induced cutaneous lesions, suggesting an active role of neutrophils in maintaining inflammation in psoriasis.


MK-329 blocks the inhibition of alcohol intake by CCK-8.
Peripheral administration of sulfated cholecystokinin octapeptide (CCK-8) potently reduces alcohol intake, preference, and blood levels in rats. MK-329 (L-364,718 or Devazepide) acts at peripheral cholecystokinin (CCKA) receptors to antagonize CCK-8's physiological and behavioral effects, such as pancreatic stimulation and inhibition of feeding. We determined whether CCKA receptor blockade would also prevent CCK-8's alcohol satiety effect. Water-deprived female and male rats (n = 7 for each) received randomized combinations of intraperitoneal injections of MK-329 (0, 100, 200, or 400 micrograms/kg) followed by CCK-8 (0 or 4 micrograms/kg). Rats were then given access to 5% w/v ethanol for 30 min, followed by 30-min access to water, with food ad lib. MK-329 at all doses significantly (p < 0.05) reduced the suppression of alcohol intake and food intake by CCK-8. MK-329 alone increased alcohol intake at 400 micrograms/kg, and increased food intake, in females and males at 100 and 200 micrograms/kg, respectively. We concluded that CCK-8's alcohol and food satiation effects depend on specific, peripheral CCKA receptors, and satiation of alcohol consumption and drinking-associated feeding reflect an endogenous functional interaction of CCK-8 with CCKA receptors.


Behavioral consequences of prenatal exposure to the organophosphate insecticide sumithion.
Prenatal administration of Sumithion 50 EC, an organophosphate insecticide, to CFY rats at 5, 10 and 15 mg/kg from days 7 to 15 of gestation, resulted in dose related decreases in open field activity and motor coordination in the offspring treated with the two higher doses. Long lasting alterations in the acquisition and extinction of a conditioned escape response, as well as increased social interactions were observed in the adult offspring. The results indicate a no effect level at 5 mg/kg under the experimental conditions in the present study.


Assessment of Risk Factors in Synanthropic and Wild Rodents Infected by Pathogenic Leptospira spp. Captured in Southern Chile.
Leptospirosis is caused by pathogenic Leptospira, and synanthropic and wildlife species of rodents are an important source of infection; however, much of the information about infection progression was obtained from murine models. The aim of this study was to assess infection status and risk factors associated with pathogenic Leptospira in synanthropic and wild rodent species and describe histopathological lesions in several organs from naturally infected animals. In a cross-sectional study, 121 rodents from three synanthropic species and two wild species were trapped in dairy farms in Southern Chile. Liver, heart, kidney, and lungs from trapped animals were fixed in formalin and stained with hematoxylin-eosin. Tissues with lesions consistent with Leptospira infection were tested by immunohistochemistry (IHC) and real-time polymerase chain reaction (qPCR) using the LipL32 antigen. Risk factors were assessed by a conditional mixed-logistic regression model. More than half (56.7%) of the negative reactors to the microscopic agglutination test were identified as infected either by IHC/qPCR. A lower risk of infection compared to the rest of the seasons was found in the fall, and the synanthropic species have a lower risk of infection in comparison with the wildlife species. IHC and qPCR contributed to the identification of pathogenic Leptospira in related histological lesions and 50% more infections than serology.


Continuous Work Support Checklist for Female Healthcare Workers: Scale Development and Validation.
Healthcare jobs are very popular among women, however in Japan, women readily quit working because of gender-role responsibilities. This study aimed to develop a workplace support checklist for women to continue to work. In 2017, we investigated 780 (female 74.8%) faculty members and healthcare professionals of one medical university in Japan. We asked them to score the extent to which they considered 35 items identified by a task team, to be related to continuous work support for female workers in healthcare. We carried out an exploratory factor analysis and extracted four domains with 16 items in all: "Support for child rearing and home care" (five items), "Information dissemination" (five items), "Active promotion of women workers to higher positions" (three items), and "Consulting and counseling service" (three items), with Cronbach's alpha values ranging from 0.88 to 0.92. We found that the first three factors were generally associated with reasonably relevant characteristics of being female, in their 30s, married, and members of faculty. We also found that women with "Intention to leave" the workplace underscored the importance of "Support for child rearing and home care" and "Consulting and counseling service". These results suggest that the checklist is reliable and valid.


Crystallization of microbial polyhydroxyalkanoates: A review.
Polyhydroxyalkanoates (PHAs), produced by the microbial fermentation, is a promising green polymer and has attracted much attention due to its excellent biocompatibility, complete biodegradability, and non-cytotoxicity. The physical properties of PHAs are closely related to their chemical and crystalline structure. Therefore, deep understanding and regulating the structure and crystallization of PHAs are the key factors to improve the performance of PHAs. This review first provides a brief overview of the development history, chemical structure, and basic properties of PHAs. Then, the crystal structure, crystal morphology, kinetics theories and crystallization behavior of nucleation-induced PHAs are systematically summarized to provide a theoretical foundation for improving PHAs crystallization rate and physical properties. In the end, the outlook on the crystallization and application prospects of PHAs is also addressed.


Extended connectivity interaction features: improving binding affinity prediction through chemical description.
Machine-learning scoring functions (SFs) have been found to outperform standard SFs for binding affinity prediction of protein-ligand complexes. A plethora of reports focus on the implementation of increasingly complex algorithms, while the chemical description of the system has not been fully exploited. Herein, we introduce Extended Connectivity Interaction Features (ECIF) to describe protein-ligand complexes and build machine-learning SFs with improved predictions of binding affinity. ECIF are a set of protein-ligand atom-type pair counts that take into account each atom's connectivity to describe it and thus define the pair types. ECIF were used to build different machine-learning models to predict protein-ligand affinities (pKd/pKi). The models were evaluated in terms of 'scoring power' on the Comparative Assessment of Scoring Functions 2016. The best models built on ECIF achieved Pearson correlation coefficients of 0.857 when used on its own, and 0.866 when used in combination with ligand descriptors, demonstrating ECIF descriptive power. Data and code to reproduce all the results are freely available at https://github.com/DIFACQUIM/ECIF. Supplementary data are available at Bioinformatics online.


Does the Modified Frailty Index (mFI-5) Predict Adverse Outcomes in Maxillofacial Fracture Repair?
Frailty has been recognized as a predictor of postoperative adverse outcomes in many surgical subspecialties. The purpose of this study was to evaluate the relationship between frailty and complications in patients undergoing operative repair of facial fractures. The authors utilized the 2011 to 2018 American College of Surgeons National Surgical Quality Improvement Program (ACS-NSQIP) databases to identify patients with facial fractures undergoing operative repair. The primary predictor variable was frailty as measured by the 5-Factor Modified Frailty Index (mFI-5). The primary outcome variable was the postoperative complication rate. Descriptive, bivariate, and multiple logistic regression statistics were utilized to evaluate the relationship between frailty and complications. During the study period, 4,290 subjects underwent operative repair of a facial fracture. Of these subjects, 4,086 (83.0%) were classified as nonfrail, 626 (12.7%) as moderately frail, and 208 (4.20%) as severely frail. A total of 237 subjects experienced a complication (4.82%), and the incidence of complications increased in a stepwise manner with increasing frailty (P ≤ .001). In multivariate regression, age (P = .050, 95% CI = 1.00 to 1.02), Native Hawaiian/Pacific Islander race (P = .018, 95% CI = 1.23 to 8.63), classification as moderately frail (P = .010, 95% CI = 1.15 to 2.66), classification as severely frail (P = .032, 95% CI = 1.06 to 3.70), mandibular fractures (P = .004, 95% CI = 1.24 to 2.98), and wound classification as contaminated (P ≤ .001, 95% CI = 1.53 to 4.57) or dirty/infected (P = .020, 95% CI = 1.16 to 5.55) were independent predictors of complications. Severely frail subjects also had greater length of hospital admission (P ≤ .001) and higher 30-day readmission rates (P ≤ .001). Frailty is an independent predictor of complications following facial fracture repair and is associated with greater length of hospital admission and 30-day readmission rates.


Contrast-enhanced agent detection imaging: Early experience in hepatocellular carcinoma.
To investigate the usefulness of contrast-enhanced Agent Detection Imaging in assessing intratumoral vasculature in hepatocellular carcinoma. Fourteen hepatocellular carcinoma nodules in 11 patients were studied with contrast-enhanced Agent Detection Imaging, a wide-band color Doppler imaging method, employing, Levovist(®), a microbubble contrast agent. High acoustic power was used with contrast-enhanced Agent Detection Imaging. Intermittent transmission of Agent Detection Imaging was performed at intervals of 200, 500, and 350 milliseconds in the early arterial phase (10 to 40 seconds), late vascular phase (1 to 3 minutes) and postvascular phase (5 to 7 minutes), respectively. The results were compared with those of three-phase dynamic CT. Intratumoral blood vessels in the early arterial phase and tumor parenchymal stain in the late vascular phase were depicted in 12 (88%) of the 14 hepatocellular carcinoma nodules, while all nodules were demonstrated as perfusion defect in the postvascular phase on contrast-enhanced Agent Detection Imaging. The results of Agent Detection Imaging, that were compared with those of dynamic CT, were all 100% : diagnostic sensitivity (12/12), specificity (2/2), and accurary (14/14). Contrast-enhanced Agent Detection Imaging is a promising method for depicting intratumoral vascularity in hepatocellular carcinoma.


Purification of human plasma fibronectin using immobilized gelatin and Arg affinity chromatography.
This protocol describes a method for purification of fibronectin (Fn) from human plasma based on a combination of gel filtration and affinity chromatography steps. Clarified plasma is first loaded onto a Sepharose CL-4B column and unbound material is sequentially purified on columns containing covalently coupled gelatin and Arg. The elution conditions are optimized to obtain a homogeneous preparation of Fn on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Although the Fn yield is expected to be lower than that obtained using other methods, affinity adsorbents based on gelatin and Arg and gentle elution steps offer advantages including a high purity of the preparation and a correctly folded protein. The preparation can be useful for interaction studies and analysis of biological and immunological activities of Fn.


Resource Use and Costs Associated to the Initial Phase of Treatment for Patients with Colorectal Cancer Receiving Post-Surgery Chemotherapy: A Cost Analysis from a Healthcare Perspective.
To estimate the resource use and costs associated to the initial phase of treatment for colorectal cancer in Iran. A retrospective study was conducted using routinely collected data within Electronic Health Records System (SEPAS), a national database representing public hospitals in Iran between March 20, 2016 and March 19, 2017. Primary end points included healthcare resource use, direct medical and non-medical costs of care in the 12-month study period. The study population included 657 patients with colorectal cancer who underwent surgery and the follow-up chemotherapy. We estimated a total direct cost of $21,407 per patient. The results indicated that direct medical costs were primarily driven by inpatient hospital care, followed by surgery, chemotherapy, and diagnostic services. The initial 12-month of treatment for colorectal cancer, including surgery and the follow-up chemotherapy, is resource intensive. The total direct costs associated to the disease are remarkable, with Inpatient hospital services being the main contributor followed by surgery and chemotherapy.


Tannic acid post-treatment of enzymatically crosslinked chitosan-alginate hydrogels for biomedical applications.
Enzyme-mediated crosslinked hydrogels as soft materials for biomedical applications have gained considerable attention. In this article, we studied the effect of tannic acid post-treatment on adhesiveness and physiochemical properties of an enzymatically crosslinked hydrogel based on chitosan and alginate. The hydrogels were soaked in TA solution at different pH (3, 5.5, 7.4, and 9) and concentrations (1, 10, 20, 30 TA wt%). Increasing the TA concentration to 30 TA wt% and pH (up to 7.4) increased the TA loading and TA release. TA post-treatment reduced the swelling ratio and degradation rate of the hydrogels due to the formation of hydrogen bonding between TA molecules, chitosan, and alginate chains resulted in higher crosslinking density. TA-reinforced hydrogels with 30 % TA (Gel-TA 30) exhibited significantly high adhesive strength (up to 18 kPa), storage modulus (40 kPa), and antioxidant activity (>96 %), antibacterial activity, and proliferation and viability of 3 T3-L1 fibroblast cells.


BMP pathway suppression is an early event in inflammation-driven colon neoplasmatogenesis of uPA-deficient mice.
The suppression of the bone morphogenetic protein (BMP) signaling pathway has been recently shown to promote adenoma-to-carcinoma transition in sporadic colon cancer. However, its role in the evolution of early preneoplastic changes to neoplasia remains elusive. In the present study, we aimed to investigate the gene expression levels of multiple extracellular BMP family constituents, including BMP ligands/receptors and inhibitors, during the early stages of inflammation-associated colon carcinogenesis. For that, we used the recently developed urokinase-type plasminogen activator (uPA)-deficient mouse model of colonic polypoidogenesis, in which adenomatous polyps arise several months after the induction of dextran sodium sulfate (DSS) colitis. In DSS-treated wild-type mice, the preneoplastic lesions which did not eventually evolve to adenomas resided in a colitic microenvironment characterized by a balanced upregulation of both BMP ligands, i.e., Bmp4/7 and BMP inhibitors, such as chordin, noggin, and gremlin-1. In the uPA-deficient tumor-promoting inflammatory microenvironment, however, there was a clear evidence for BMP pathway suppression. By contrast to DSS-treated wild-type controls, the inflammation-associated Bmp4 upregulation was abolished, and the BMP signaling suppression was further enhanced by a particularly high increase of gremlin-1 expression. These findings propose that BMP pathway suppression in colon cancer could be associated with very early stages of the preneoplasia-to-neoplasia sequence of events.


Comparative changes in treatment practices and clinical outcomes following implementation of a prospective payment system: the STEPPS study.
The aim of the US dialysis Prospective Payment System bundle, launched in January 2011, was reduction and more accurate prediction of costs of services, whilst maintaining or improving patient care. Dialysis facilities could either adopt the bundle completely (100%) in the first year of launch, or phase-in (25%) over four years. Differences in practice patterns and patient outcomes were hypothesized to occur in facilities that phased-in 25% compared to those that did not. Data are from STEPPS, a study of 51 small dialysis organization facilities designed to describe trends in dialytic treatment before and after bundle implementation. Baseline was defined as October-December 2010; follow-up as January-December 2011. Facility- and patient-level data were collected at enrollment and regularly thereafter. Cox proportional hazards and linear multi-level models were used to estimate the effect of opting-in 25% (vs. 100%) on practice patterns and clinical outcomes. 12 facilities (patient n = 346) opted-in 25% and 37 facilities (patient n = 1296) opted-in 100% to the dialysis bundle. At baseline, patients at 25% facilities were primarily covered by Medicare, were more likely to be black, and were receiving higher monthly epoetin alfa (EPO) doses. Throughout 2011, patients in 100% facilities received lower monthly EPO doses, and had lower mean hemoglobin concentrations; hospitalization and mortality rates were numerically lower in 25% facilities but not statistically different. The economic pressure for dialysis providers to work within an expanded composite rate bundle whilst maintaining patient care may be a driver of practice indicator outcomes. Additional investigations are warranted to more precisely estimate clinical outcomes in patients attending facilities enrolling into the bundle 100% relative to the previous fee-for-service framework.


Correction: Gravely, S., et al. Awareness, Trial, and Current Use of Electronic Cigarettes in 10 Countries: Findings from the ITC Project. Int. J. Environ. Res. Public Health 2014, 11, 11691-11704.
The authors wish to make the following amendments to their paper published in International Journal of Environmental Research and Public Health.


Implementation of a faith-based physical activity intervention: insights from church health directors.
Churches are becoming increasingly popular settings for conducting health promotion programs. Retrospective interviews were conducted with 19 health directors from churches taking part in the evaluation of a large-scale faith-based physical activity initiative. This paper first describes program implementation, church leadership support, and changes in church leadership, and then relates these variables to program outcomes (percentage of participants meeting physical activity recommendations). Finally, barriers and successes to program implementation are reported. The most commonly reported intervention activities implemented by churches were bulletin boards related to healthy eating and physical activity (79%) followed by bulletin inserts (69%), walking programs (58%), chair exercises (48%), praise aerobics (27%), a 10-min exercise CD (26%), and an 8 week behavior change class (26%). Significant increases in physical activity were associated with churches which had ever implemented the behavior change class at the 1-year follow-up. According to health directors, pastors sometimes to often spoke about physical activity and diet from the pulpit but rarely to sometimes took part in program activities. They also reported that pastors spouses' never to rarely spoke about physical activity and diet from the pulpit, and rarely to sometimes took part in program activities. About 68% of the churches had at least one change in pastor over the 3-year study. A majority of these variables, however, were not related to changes in physical activity. Potential reasons for these lack of associations are discussed.


Inflammatory cell distribution in colon mucosa as a new tool for diagnosis of irritable bowel syndrome: A promising pilot study.
Currently, there are no histological criteria to diagnose irritable bowel syndrome (IBS). Our aims were (i) to examine the distribution of inflammatory cells in the colon of healthy and IBS subjects and (ii) to find histological diagnosis criteria for IBS. Colonic biopsies were taken from four distinct regions of the colon from 20 controls (HC) and 11 patients with IBS (4 with constipation (IBS-C) and 7 with diarrhea (IBS-D) and embedded in paraffin. Macrophages, mast cells, eosinophils, and T lymphocytes were immunostained and positive cells counted. In both HC and IBS patients, global cellularity decreased from the cecum to the rectum (P < .01) which is attributed to reduced number of macrophages (P < .05) and eosinophils (P < .001) but not T cells. Mast cells were reduced in IBS (P < .05) but not in HC, particularly in IBS-D (P < .05). Results showed higher number of macrophages in the left colon of IBS subjects than HC (P < .05). Here we report a decreasing gradient of immune cells from the cecum to the rectum of the human colon. Although global cellularity cannot be used to distinguish between IBS and HC, closer analysis of macrophages and mast cells may be useful markers to confirm IBS histologically and to differentiate between IBS-C and IBS-D when clinical presentation alternates between constipation and diarrhoea. This pilot study remains to be confirmed with greater number of patients.


Connections of the superior vestibular nucleus with the oculomotor and red nuclei in the rat: an electron microscopic study.
Phaseolus vulgaris leucoagglutinin (PHA-L) was injected into the individual vestibular nuclei of the rat to study their efferent connections. One of the major differences between the connections of these nuclei was found at the level of the mesencephalon: the eye-moving cranial nerve nuclei received the densest projection from the superior vestibular nucleus (SVN). In the present electron microscopic study, we have found that terminals of SVN origin established symmetric synaptic contacts in the oculomotor nucleus. More than two-thirds of PHA-L-labeled boutons terminated on dendrites, the rest of them established axosomatic contacts. Most of the labeled terminals were GABA-positive, supporting the results of previous physiological experiments, which showed inhibitory effects. In the mesencephalon, the other termination area was found in the red nucleus. The PHA-L-labeled boutons of SVN origin were in close contact with the perikarya and proximal dendrites of the magnocellular part of the red nucleus. The types of synaptic contacts and distribution of terminals of SVN origin were similar to those found in the oculomotor nucleus. Our results indicate that the SVN can modify the activity of the cerebellorubral and corticorubral pathways, exerting inhibitory action on the neurons of the red nucleus.


The impact of continuous use of lumbosacral orthoses on trunk motor performance: a systematic review with meta-analysis.
Lumbosacral orthosis (LSO) is prescribed by general practitioners for the management of low back pain. It may be speculated that continuous use of an LSO for a prolonged period reduces mechanical loading to the trunk muscle in daily living and results in impairments of the trunk muscle. This study aims to investigate whether trunk motor performances are impaired by the continuous use of an LSO. This is a systematic review with meta-analysis. A systematic search was undertaken using PubMed, EMBASE, MEDLINE, CINAHL, SCOPUS, and Cochrane Library from inception to November 2016. Inclusion criteria were (1) the use of an LSO for ≥2 days, (2) the use of a soft LSO designed for musculoskeletal conditions, (3) absence of cointervention except education, and (4) measures of trunk motor performance. The following were excluded: (1) studies with insufficient data and (2) studies with poor methodological quality (<9/16) in the modified McMaster Critical Review Form for Quantitative Studies. The Grading of Recommendations Assessment, Development and Evaluation (GRADE) system was used to determine the quality of evidence. Data of eight studies were analyzed. The most common measures for motor performances were the maximum strength of the trunk flexors and extensors and the endurance and fatigability of the trunk extensors. In all measures, 95% confidence intervals of the pooled standardized mean difference between the control or preintervention group and the intervention or postintervention group included zero. Further, quality of evidence ranged from low to very low in the GRADE system in all findings of the meta-analyses. The meta-analyses demonstrated no negative effect by the continuous use of an LSO for 1-6 months. However, the quality of evidence ranged from low to very low, and more high-quality trials are required to draw a definitive conclusion on the impact of the continuous use of an LSO on trunk motor performances.


Prenatal cocaine produces deficits in serotonin mediated neuroendocrine responses in adult rat progeny: evidence for long-term functional alterations in brain serotonin pathways.
Cocaine produces biochemical alterations in brain serotonin (5-HT) neurons. Since 5-HT is critical to the development of fetal 5-HT neurons and target tissues, we hypothesized that in utero exposure to cocaine could result in long-term alterations in postnatal 5-HT systems. Pregnant Sprague-Dawley rats were administered either saline or (-)cocaine (15 mg/kg, s.c., b.i.d.) from gestational day 13 to 20. Prenatal cocaine exposure did not alter litter size, gender number, or progeny birth weights. Functional alterations in serotonergic systems were determined in postnatal day (PD) 70 male progeny by measuring changes in 5-HT mediated plasma hormones following a single 8 mg/kg injection of the 5-HT releaser p-chloroamphetamine (PCA). Cocaine exposed male progeny exhibited significant reductions in adrenocorticotropic hormone (ACTH, -43%) and renin (-62%) responses to PCA. However, no alterations were observed in the corticosterone or prolactin response to PCA. In utero exposure to cocaine did not alter basal levels of ACTH, renin, corticosterone, or prolactin. There were no significant differences in the density of either hypothalamic or cortical 5-HT uptake sites. Likewise, there were no significant differences in the densities of any of the 5-HT1 receptor subtypes or in the density of 5-HT2 receptors in cortex. These data, which provide the first demonstration of deficits in 5-HT mediated neuroendocrine function in adult progeny following in utero exposure to cocaine, indicate long-term functional alterations of brain 5-HT systems.


Spatiotemporal regulation of Ras activity provides directional sensing.
Cells' ability to detect and orient themselves in chemoattractant gradients has been the subject of numerous studies, but the underlying molecular mechanisms remain largely unknown [1]. Ras activation is the earliest polarized response to chemoattractant gradients downstream from heterotrimeric G proteins in Dictyostelium, and inhibition of Ras signaling results in directional migration defects [2]. Activated Ras is enriched at the leading edge, promoting the localized activation of key chemotactic effectors, such as PI3K and TORC2 [2-5]. To investigate the role of Ras in directional sensing, we studied the effect of its misregulation by using cells with disrupted RasGAP activity. We identified an ortholog of mammalian NF1, DdNF1, as a major regulator of Ras activity in Dictyostelium. We show that disruption of nfaA leads to spatially and temporally unregulated Ras activity, causing cytokinesis and chemotaxis defects. By using unpolarized, latrunculin-treated cells, we show that tight regulation of Ras is important for gradient sensing. Together, our findings suggest that Ras is part of the cell's compass and that the RasGAP-mediated regulation of Ras activity affects directional sensing.


An IgM-like inhalable ACE2 fusion protein broadly neutralizes SARS-CoV-2 variants.
Many of the currently available COVID-19 vaccines and therapeutics are not effective against newly emerged SARS-CoV-2 variants. Here, we developed the metallo-enzyme domain of angiotensin converting enzyme 2 (ACE2)-the cellular receptor of SARS-CoV-2-into an IgM-like inhalable molecule (HH-120). HH-120 binds to the SARS-CoV-2 Spike (S) protein with high avidity and confers potent and broad-spectrum neutralization activity against all known SARS-CoV-2 variants of concern. HH-120 was developed as an inhaled formulation that achieves appropriate aerodynamic properties for rodent and monkey respiratory system delivery, and we found that early administration of HH-120 by aerosol inhalation significantly reduced viral loads and lung pathology scores in male golden Syrian hamsters infected by the SARS-CoV-2 ancestral strain (GDPCC-nCoV27) and the Delta variant. Our study presents a meaningful advancement in the inhalation delivery of large biologics like HH-120 (molecular weight (MW) ~ 1000 kDa) and demonstrates that HH-120 can serve as an efficacious, safe, and convenient agent against SARS-CoV-2 variants. Finally, given the known role of ACE2 in viral reception, it is conceivable that HH-120 has the potential to be efficacious against additional emergent coronaviruses.


Microvascular spasm is mediated by vasopressin fibers in the rat hippocampal slice.
Microvessels in the rat hippocampal slice can be used as an in vitro model for the study of cerebral vasospasm. Serum from coagulated blood causes prompt and long-lasting microvascular constriction that is neurogenic in nature. Here we show that a candidate spasmogen, thromboxane B2, has excitatory action on neural elements in the slice. However, spasmogenic serum lacks this excitatory effect. Instead, it is inhibitory for a major population of slice neurons. Thus, neurogenic microvascular spasm is produced by a subpopulation of slice neurons or projection fibers, not all neurons acting in concert. Arginine vasopressin (AVP) is contained in fibers that project to the hippocampus, and hippocampal microvessels contain pressor (V1) receptors for the peptide. AVP causes vasoconstriction in the slice, and a specific V1 antagonist for the peptide blocks the microvascular spasm induced by blood serum. The results are interpreted to mean that neurogenic microvascular spasm is mediated by locally released AVP.


Pro-tobacco influences and susceptibility to smoking cigarettes among middle and high school students--United States, 2011.
Smoking is a leading cause of cancer, and most smokers begin during adolescence. We examined the proportion of adolescents exposed to pro-tobacco advertising and assessed the association between this exposure and susceptibility to smoking. Data from the 2011 National Youth Tobacco Survey were used to calculate the proportion of susceptible middle school (MS) and high school (HS) students exposed to pro-tobacco advertisements through stores, magazines, and the Internet. Following previous work, susceptibility to smoking cigarettes was defined as "never smoked but open to trying cigarettes." In 2011, 81.5% of MS students and 86.9% of HS students were exposed to tobacco advertisements in stores; 48.2% of MS students and 54.0% of HS students were exposed to such advertising in magazines. Exposure to tobacco advertisements on the Internet was similar for MS (40.8%) and HS students (40.2%). Of those surveyed, 22.5% of MS students and 24.2% of HS students were susceptible to trying cigarettes. Exposure to magazine advertising declined from 71.8% in 2000 to 46.1% in 2009 among susceptible MS students; however, exposure increased to 55.4% in 2011. Tobacco advertising seen through the Internet among susceptible HS students increased from 25.9% in 2000 to 44.7% in 2011. Adolescents continue to be exposed to pro-tobacco advertisements. Adolescents susceptible to smoking are more likely to report exposure to pro-tobacco advertisements. In addition to continued monitoring, more effective interventions to eliminate youth exposure to pro-tobacco marketing are needed.


Functional status following injury: what recovery pathways do people follow?
Functional status can be affected for considerable time after injury. Individual's functional status trajectories, or pathways, following injury may provide insights into achieving, or not achieving, optimal functional status. This study aims to (1) investigate functional status trajectories of injured individuals over two years by multiple dimensions and, (2) determine whether there are differences in functional status trajectories between those hospitalised and non-hospitalised. Data from the Prospective Outcomes of Injury Study, a longitudinal cohort (n=2856) of injured New Zealanders, was used. Functional status was assessed using the EQ-5D (plus a cognitive dimension) at 3, 12 and 24 months post injury. For each dimension (mobility, self-care, usual activities, pain/discomfort anxiety/depression and cognition), individual-level trajectories were created based on whether participants had attained (or exceeded) their pre-injury functional status at each time-point. Participants had varied pathways to attaining their pre-injury functional status which was not revealed by cross-sectional group-level data. When all dimensions were considered together, 24% of participants attained their pre-injury functional status but did not maintain it at a subsequent phase. By EQ-5D dimension, this varied from 5% (self-care) to 22% (pain/discomfort). Twenty-six percent of non-hospitalised participants attained, but did not maintain, their pre-injury status compared to 18% of those hospitalised. Cross-sectional group-level prevalence does not adequately depict the underlying pathways experienced by individual participants. Our analyses indicate the importance of following up all study participants in longitudinal studies, including those reporting to have attained 'recovery' and of not under-estimating the impact of non-hospitalised injuries.


Magnetic resonance imaging and the diagnosis of avascular necrosis of the femoral head.
Low back pain and/or pain radiating to one or both lower limbs can result from causes other than intervertebral disc disease. Recently three patients presented with low back pain radiating down the legs. When CT examination of the lumbar spine proved unrewarding, magnetic resonance imaging (MRI) was performed. Avascular necrosis (AVN) of the femoral head was diagnosed and subsequently treated before femoral head collapse occurred. In patients with unexplained low back pain, AVN of the hip should be considered and ruled out by MRI, even if conventional plain films, polytomes, and radionuclide scans are negative. The early diagnosis of AVN of the femoral head may be critical in determining whether a surgical revascularization procedure might be successful in treatment, or replacement with an endoprosthesis will be necessary, if femoral head collapse has already occurred.


Osteoporosis screening using machine learning and electromagnetic waves.
Osteoporosis is a disease characterized by impairment of bone microarchitecture that causes high socioeconomic impacts in the world because of fractures and hospitalizations. Although dual-energy X-ray absorptiometry (DXA) is the gold standard for diagnosing the disease, access to DXA in developing countries is still limited due to its high cost, being present only in specialized hospitals. In this paper, we analyze the performance of Osseus, a low-cost portable device based on electromagnetic waves that measures the attenuation of the signal that crosses the medial phalanx of a patient's middle finger and was developed for osteoporosis screening. The analysis is carried out by predicting changes in bone mineral density using Osseus measurements and additional common risk factors used as input features to a set of supervised classification models, while the results from DXA are taken as target (real) values during the training of the machine learning algorithms. The dataset consisted of 505 patients who underwent osteoporosis screening with both devices (DXA and Osseus), of whom 21.8% were healthy and 78.2% had low bone mineral density or osteoporosis. A cross-validation with k-fold = 5 was considered in model training, while 20% of the whole dataset was used for testing. The obtained performance of the best model (Random Forest) presented a sensitivity of 0.853, a specificity of 0.879, and an F1 of 0.859. Since the Random Forest (RF) algorithm allows some interpretability of its results (through the impurity check), we were able to identify the most important variables in the classification of osteoporosis. The results showed that the most important variables were age, body mass index, and the signal attenuation provided by Osseus. The RF model, when used together with Osseus measurements, is effective in screening patients and facilitates the early diagnosis of osteoporosis. The main advantages of such early screening are the reduction of costs associated with exams, surgeries, treatments, and hospitalizations, as well as improved quality of life for patients.


Presence of GnRH3 in sturgeon and the roles of GnRH1 and GnRH2 on the regulation of LH/FSH in mature female sterlet Acipenser ruthenus in vitro.
Three forms of gonadotropin-releasing hormones (GnRHs), ArGnRH1, ArGnRH2, and ArGnRH3, were identified in sterlet. Compared with their orthologue, ArGnRH1 and ArGnRH2 have conserved core decapeptide but show low identity in the signal peptide and the rest of the sequences. The existence of the GnRH3 paralogue of sturgeon was predicted for the first time with TBLASTN by using the amino acid sequences of catshark and whale shark GnRH3 precursor as queries against the whole genome and transcript data of sterlet. The predicted ArGnRH3 cDNA sequence was composed of three exons containing all the elements of the GnRH family. The successful molecular cloning of GnRH3 from sterlets verified its expression in the brain of sturgeons. The analysis of the ArGnRH3 amino acid sequence revealed a completely conserved decapeptide sequence that shows 100% identity with the sequence of teleosts and differs in one amino acid with that of the cartilaginous fish (catshark and whale shark) at the 5th position. The structure of the phylogenetic tree showed that a total of 52 vertebrate GnRH sequences were clustered into three main clades corresponding to GnRH1, GnRH2, and GnRH3. The ArGnRH3 sequence is the oldest GnRH3 identified in teleosts. The tissue distribution analysis showed that ArGnRH1 was expressed in all the 13 examined tissues of females and in most of the tested tissues of male fish, with the highest expression in the pituitary and hypothalamus. ArGnRH2 is only expressed in the pituitary, hypothalamus, and gonads of both female and male sterlets. ArGnRH3 mRNA could be detected in the pituitary, hypothalamus, and gonad in both female and male fish. It is also present in the spleen, head kidney, and gill in female fish and in kidney and heart in male fish. However, the ArGnRH3 only showed weak expression in all the positive tissues. ArGnRH1 and ArGnRH2 active decapeptides were synthesized to investigate their roles on the regulation of LH/FSH using a mixed brain cell line from a sexually mature female sterlet. The results showed that ArGnRH1 and ArGnRH2 exerted different effects on the gene expression and release of gonadotropins. ArGnRH1 promoted the expression of fshβ significantly around 48 h, and the expression was suppressed when the treatment time was extended to 72 h. ArGnRH1 had no significant effects on the level of either mRNA or secreted lh in any of the tested treatment length or concentrations. Moreover, ArGnRH1 did not stimulate the activity of gonadotropins in the maturation stage of female sturgeons. ArGnRH2 promoted the expression of fshβ at 24 h and 48 h and increased mRNA level of lhβ at 6 h and 48 h, accompanied by the significant secretion of LH at 72 h, although the high mRNA level of fsh did not correlate with the secretion of FSH in ArGnRH2-treated groups. In conclusion, ArGnRH2 plays an important role in the maturation stage of female sterlets. Therefore, ArGnRH2 has the potential to induce ovulation and spermiation in sturgeons.


Inhibition of apoptosis by menadione on exposure to UVA.
Quinones are widely distributed in the environment, both as natural products and as pollutants. This paper reports that one of the simplest quinones, 2-methyl-1,4-naphthoquinone (menadione), effectively inhibited apoptosis in the presence of UVA. Menadione suppressed the apoptosis induced by serum depletion and cell detachment. This effect was significantly enhanced by UVA irradiation. An antioxidant, N-acetylcysteine, completely inhibited the antiapoptotic effects of both menadione itself and menadione plus UVA, and peroxidation of the cells after treatment was observed using a probe to detect the intracellular production of peroxides. By contrast, 2-hydroxy-1,4-naphtoquinone (lawsone) showed no antiapoptotic effect in the presence or absence of UVA. Lawsone is reported not to undergo the redox process that produces reactive oxygen species. These results indicated that intracellular peroxidation contributed to the antiapoptotic effects of both menadione itself and menadione plus UVA. Dysregulation of the apoptotic process is critical to carcinogenesis. The photosensitization of quinone compounds as it relates to the inhibition of apoptosis should be examined in the future.


Nursing stakeholder identification guidelines for human resources for health and health workforce development: A scoping review.
To investigate the existence of guidelines on the identification of nursing stakeholders as part of planning for human resources for health processes. Effective involvement of nursing stakeholders in planning and implementing human resources for health policies is strongly advocated by leading global bodies. Systematic identification of nursing stakeholders at an early stage is fundamentally important. Guidelines to support appropriate identification and inclusion of nursing stakeholders could support the active involvement of nurses and midwives in human resources for health planning processes at all levels. We conducted a scoping review using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for scoping reviews. We conducted a widely inclusive search for all types of records, including searches of bibliographic databases (PubMed, CINAHL, Scopus and Web of Science) and manual searches of selected websites and internet archives to identify grey literature, published in English since 2009. Search terms related to guidelines, stakeholder engagement and the health workforce. Of the 1058 potentially relevant sources identified, two studies met inclusion criteria. Both were guidelines produced by global bodies more than 12 years ago. Cochrane guidance on reporting 'near-empty' reviews was followed, and eight additional sources meeting most of the inclusion criteria were identified and critiqued. Guidelines regarding the process of nursing stakeholder identification specific to human resources for health planning processes are scarce and require updating. Critique of recent practices suggests considerable methodological variety and sub-optimal identification of nursing stakeholders. Nursing stakeholder engagement is an essential component of human resources for health planning processes, and the gap in literature points to a need for up-to-date guidance to ensure nurses' active involvement.


Effect of the C1q on the soluble collagen-platelet interaction.
The mechanism by which C1q inhibited soluble collagen-induced platelet aggregation was examined. Platelet aggregation induced by soluble collagen in gel filtrated platelets was inhibited by the addition of C1q. There were no cross-reactions between C1q, purified soluble collagen receptor and both of their polyclonal antibodies in enzyme-linked immunosorbent assays. These results suggest that the effect of C1q on soluble collagen-induced platelet aggregation does not compete at the same binding site on the platelet surface.


Birth prevalence of homocystinuria in Central Europe: frequency and pathogenicity of mutation c.1105C>T (p.R369C) in the cystathionine beta-synthase gene.
To estimate the frequency of the cystathionine beta-synthase deficiency caused by c.1105C>T mutation in Central Europe compared to Norway, and to examine the pathogenicity of the corresponding p.R369C mutant enzyme. Mutation c.1105C>T was analyzed in 600 anonymous Czech newborn blood spots. Catalytic activity and quaternary structure of the p.R369C mutant was evaluated after expression in 2 cellular systems. Population frequency of the c.1105C>T mutation was 0.005, predicting the birth prevalence of homocystinuria of 1:40000, which increased to 1:15500 in a model including 10 additional mutations. In Escherichia coli the p.R369C mutant misfolded, and its activity was severely reduced, and expression in Chinese hamster ovary cells enabled proper folding with activity decreased to 63% of the wild-type enzyme. This decreased activity was not due to impaired K(m) for both substrates but resulted from V(max) lowered to 55% of the normal cystathionine beta-synthase enzyme. The c.1105C>T (p.R369C) allele is common also in the Czech population. Although the p.R369C mutation impairs folding and decreases velocity of the enzymatic reaction, our data are congruent with rather mild clinical phenotype in homozygotes or compound heterozygotes carrying this mutation.


Occurrence of an active regulatory mechanism of protein synthesis during starvation and refeeding in Bombyx mori fat body.
The origin of the amino acids which participate in protein synthesis at the recovery from starvation have been determined in the fat body from Bombyx mori larvae. Endogeneous amino acids have been labelled with [3H] leucine and ingested ones with [14C] leucine, allowing their discrimination in the organism. 22 minutes after refeeding, proteosynthetic activity of the fat body, estimated by the polysome level, is increased 2.5 fold. Endogeneous leucine represents more than 90 p. cent of the leucine present in nascent polypeptides. Free leucine pools of the fat body and of hemolymph increase, mainly through the release of endogeneous leucine. It is therefore concluded that refeeding with amino acids induces the production of a signal or critical factor, responsible for the increase in proteosynthetic activity in the fat body.


Canonical modeling of anticipatory vaccination behavior and long term epidemic recurrence.
Vaccination is one of humanity's main tools to fight epidemics. In most countries and for most diseases, vaccination is offered on a voluntary basis. Hence, the spread of a disease can be described as two interacting opposite dynamic systems: contagion is determined by past vaccination, while individuals decide whether to vaccinate based on beliefs regarding future disease prevalence. In this study, we show how the interplay between such anticipating behavior and the otherwise biological dynamics of a disease may lead to the emergence of recurrent patterns. We provide simulation results for (i) a Measles-like outbreak, (ii) canonical fully rational and far-sighted individuals, (iii) waning vaccine efficacy and vital dynamics, and (iv) long periods of time, i.e. long enough to observe several vaccination peaks. For comparison, we conducted a similar analysis for individuals with adaptive behavior. As an extension, we investigated the case where part of the population has an anti-vaccination stance.


Detection and characterization of merohedral twinning in crystals of oxalyl-coenzyme A decarboxylase from Oxalobacter formigenes.
Oxalyl-coenzyme A decarboxylase is a thiamin diphosphate dependent enzyme active in the catabolism of the highly toxic compound oxalate. The enzyme from Oxalobacter formigenes has been expressed as a recombinant protein in Escherichia coli, purified to homogeneity and crystallized. Two crystal forms were obtained, one showing poor diffraction and the other merohedral twinning. Crystals in the former category belong to the tetragonal space group P4(2)2(1)2. Data to 4.1 A resolution were collected from these crystals and an incomplete low resolution structure was initially determined by molecular replacement. Crystals in the latter category were obtained by co-crystallizing the protein with coenzyme A, thiamin diphosphate and Mg(2+)-ions. Data to 1.73 A were collected from one of these crystals with apparent point group 622. The crystal was found to be heavily twinned, and a twin ratio of 0.43 was estimated consistently by different established methods. The true space group P3(1)21 was deduced, and a molecular replacement solution was obtained using the low resolution structure as template when searching in detwinned data.