Spontaneous generation and survival of blood dendritic cells in mononuclear cell culture without exogenous cytokines.
Studies on purified blood dendritic cells (DCs) are hampered by poor viability in tissue culture. We, therefore, attempted to study some of the interactions/relationships between DCs and other blood cells by culturing unseparated peripheral blood mononuclear cell (PBMC) preparations in vitro. Flow cytometric techniques were used to undertake a phenotypic and functional analysis of DCs within the cultured PBMC population. We discovered that both the CD11c(+) and CD11c(-) CD123(hi) DC subsets maintained their viability throughout the 3-day culture period, without the addition of exogenous cytokines. This viability was accompanied by progressive up-regulation of the surface costimulatory (CD40, CD80, CD86) and activation (CMRF-44, CMRF-56, CD83) molecules. The survival and apparent production of DCs in PBMC culture (without exogenous cytokines) and that of sorted DCs (with cytokines) were evaluated and compared by using TruCOUNT analysis. Absolute DC counts increased (for CD123(hi) and CD11c(+) subsets) after overnight culture of PBMCs. Single-cell lineage depletion experiments demonstrated the rapid and spontaneous emergence of "new" in vitro generated DCs from CD14(+)/CD16(+) PBMC radioresistant precursors, additional to the preexisting ex vivo DC population. Unlike monocyte-derived DCs, blood DCs increased dextran uptake with culture and activation. Finally, DCs obtained after culture of PBMCs for 3 days were as effective as freshly isolated DCs in stimulating an allogeneic mixed leukocyte reaction.


Organometallic cobalamin anticancer derivatives for targeted prodrug delivery via transcobalamin-mediated uptake.
Herein we report the synthesis of new water-soluble vitamin B12 prodrugs bearing metal complexes at the β-upper side of the cobalt center. A total of three derivatives with the general design {Co-C[triple bond, length as m-dash]C-bpy-M}, where M represents a cytotoxic metal complex, were prepared and tested for their cytotoxicity against MCF-7 breast cancer cells. The choice of the metal was oriented on the eminent Pt and promising Ru and Re species to demonstrate the general applicability of the approach. The recognition of the derivatives by transcobalamin was demonstrated by competitive displacement assays using rhodamine labeled B12. This compound further served to prepare a dual luminescent probe by orthogonal synthesis with M = ((HCCbpy)Ru(bpy)2)Cl2 and to perform in vitro assays. Cellular imaging experiments allowed us to observe the different compartmentalization of both dyes and thus prove that the species follow the natural cobalamin uptake as well as the self-triggered release of the β-upper complex.


Complement C7 (C7), a Potential Tumor Suppressor, Is an Immune-Related Prognostic Biomarker in Prostate Cancer (PC).
Objectives: Prostate cancer (PC) is the second most frequent tumor in men, which has a high recurrence rate and poor prognosis. Therefore, this study aimed to identify novel prognostic biomarkers and therapeutic targets for immunotherapy and small molecule drugs for PC treatment. Materials and Methods: The Estimation of Stromal and Immune cells in Malignant Tumor tissues using Expression data (ESTIMATE) algorithm was applied to calculate immune scores and stromal scores of TCGA-PRAD data. Differentially expressed genes (DEGs) were identified using R package "limma." GO, KEGG, and DO analyses were performed to analyze DEGs. Overall survival and disease-free survival analyses were conducted for hub gene identification. To validate the hub gene at the mRNA and protein expression levels, genetic alterations were measured, and CCLE and Cox regression analyses were performed. Connectivity map (CMap) analysis and GSEA were performed for drug exploration and function analysis, respectively. Results: Immune scores ranged from -1795.98 to 2339.39, and stomal scores ranged from -1877.60 to 1659.96. In total, 45 tumor microenvironment (TME)-related DEGs were identified, of which Complement C7 (C7) was selected and validated as a hub gene. CMap analysis identified six small molecule drugs as potential agents for PC treatment. Further analysis demonstrated that C7 expression was significantly correlated with clinical T, pathological N, and immune infiltration level. Conclusions: In conclusion, of the 45 TME-related DEGs, C7 was shown to correlate with PC prognosis in patients, indicating it as a novel prognostic biomarker and immunotherapy target in PC. Additionally, six small molecule drugs showed strong therapeutic potential for PC treatment.


Influence of minor trauma on surgical results in patients with cervical OPLL.
This study was designed to evaluate the relationship between minor trauma and surgical results of cervical ossification of the posterior longitudinal ligament (OPLL), focusing on static compressive and dynamic factors contributing to cord compression plus the type of attendant OPLL. Of the 91 patients treated surgically, 26 had sustained minor trauma (injury group), and the other 65 had no trauma (noninjury group). The pre-/postoperative JOA scores and recovery ratio for the injury group were significantly lower than those for the noninjury group. In the segmental, localized, and mixed types, which had good ranges of motion of the cervical spine, the recovery ratios of the injury group were significantly lower than those of the noninjury group. In the continuous type, which had a poor range of motion and severe spinal cord compression, there was no significant difference in the recovery ratio between the injury and noninjury groups. Thus, dynamic factors may play an important role in neurologic deterioration after minor trauma in patients with segmental, mixed, and localized cervical OPLL.


Surrogate markers of insulin resistance: A review.
Insulin resistance is a hallmark of obesity, diabetes, and cardiovascular diseases, and leads to many of the abnormalities associated with metabolic syndrome. Our understanding of insulin resistance has improved tremendously over the years, but certain aspects of its estimation still remain elusive to researchers and clinicians. The quantitative assessment of insulin sensitivity is not routinely used during biochemical investigations for diagnostic purposes, but the emerging importance of insulin resistance has led to its wider application research studies. Evaluation of a number of clinical states where insulin sensitivity is compromised calls for assessment of insulin resistance. Insulin resistance is increasingly being assessed in various disease conditions where it aids in examining their pathogenesis, etiology and consequences. The hyperinsulinemic euglycemic glucose clamp is the gold standard method for the determination of insulin sensitivity, but is impractical as it is labor- and time-intensive. A number of surrogate indices have therefore been employed to simplify and improve the determination of insulin resistance. The object of this review is to highlight various aspects and methodologies for current and upcoming measures of insulin sensitivity/resistance. In-depth knowledge of these markers will help in better understanding and exploitation of the condition.


8-hydroxydeoxyguanosine as a useful marker for determining the severity of trichloroethylene exposure.
The aim of this study was to determine 8-hydroxydeoxyguanosine (8-OH-dG) levels in trichloroethylene (TCE)-exposed workers. Oxidative stress biomarkers and biochemical parameters were monitored among 26 TCE-exposed workers and 78 age-matched control subjects. Levels of urinary 8-OH-dG were analyzed by liquid chromatography tandem mass spectrometry. Urinary 8-OH-dG levels were significantly higher for TCE-exposed group (p < .001). Spearman's correlation test revealed positive correlations between urinary trichloroacetic acid levels and age, urinary 8-OH-dG, urinary total oxidant status, and urinary total antioxidant status (p = .042, p < .001, p < .001, and p < .001, respectively). 8-OH-dG may be a useful marker to determine the extent of TCE exposure.


Can child deaths be prevented? The Arizona Child Fatality Review Program experience.
To determine the causes and preventability of child deaths; to assess the accuracy of death certificate information; and to assess the number of child abuse deaths that are misdiagnosed as deaths attributable to natural or accidental causes. Analysis of deaths of children <18 years old that occurred between 1995-1999 using the data collected by the Arizona Child Fatality Review Program (ACFRP). From 1995-1999, local multidisciplinary child fatality review teams (CFRTs) have reviewed 95% of all deaths of children <18 years old in Arizona. Each team has access to the child's death certificate, autopsy report, hospital records, child protective services records, law enforcement reports, and any other relevant documents that provide insight into the cause and preventability of a child's death. After reviewing these documents, the team determines the cause of death, its preventability, and the accuracy of the death certificate. The ACFRP defines a child's death as preventable if an individual or the community could reasonably have done something that would have changed the circumstances that led to the child's death. The ACFRP determined that 29% (1416/4806) of these deaths could have been prevented, and preventability increased with the age of the child. Only 5% (81/1781) of neonatal deaths were considered preventable, whereas the deaths of 38% of all children older than 28 days were considered preventable. By 9 years of age, the majority of child deaths (56%) were considered preventable. Deaths attributable to medical conditions were far less likely to be considered preventable than deaths attributable to unintentional injuries. Although 62% of all deaths in Arizona during the 5-year period were attributable to medical conditions, only 8% (253/2983) of these deaths were considered preventable. In contrast, 91% (852/934) of the deaths attributable to unintentional injuries were considered preventable. Motor vehicle crashes accounted for 634 of the deaths resulting from injuries, and drowning accounted for 187 deaths. Motor vehicle crashes were the leading cause of death for all children in Arizona over 1 year of age. Only 18% of child passengers and 3% of adolescent drivers who died were known to be appropriately restrained. The typical drowning victim was a young child who drowned in the family's backyard pool. Indeed, 70% (131/187) of the drowning victims were <5 years old, and 62% (81/131) of these children died in a backyard pool. Supervision of the child and pool fencing could have prevented 90% of these deaths. Most deaths attributable to medical conditions occurred in the first year of life. Prematurity was the most common medical condition (1036 deaths) followed by congenital anomalies (662 deaths) and infectious diseases (470 deaths). Some of the reasons why CFRTs believed a medical death was preventable included inadequate emergency medical services, poor continuity of care, and delay in seeking care because of lack of health insurance. There were 4 deaths resulting from infections that were vaccine-preventable. There were 263 deaths attributable to sudden infant death syndrome. Only 38 of these infants were found lying on their back; 35 were found lying on their side. The death rate from sudden infant death syndrome decreased from 1.1 per 1000 infants <1 year of age in 1995 to 0.5 in 1999. There were 33 deaths that the CFRTs concluded were attributable to unsafe sleeping arrangements that resulted in unintentional suffocation. From 1995-1999, 317 Arizona children died from gun shot wounds. Most of these deaths were homicides (175) or suicides (109). All suicide deaths occurred in children >9 years old, and 77% of these children were >14 years old. The typical suicide victim was male (83%) and used a gun (70%) to kill himself. After review by the CFRTs, it was determined that 5 of the 67 child abuse deaths were misdiagnosed as attributable to natural or accidental causes on the death certificate. In 3 of these 5 cases, the child was in a persistent vegetative state and died many years after the episode of child abuse. Although inaction or inappropriate action by Child Protective Services (CPS) is often thought to be the cause of child abuse deaths, the ACFRP determined that in 79% of child abuse deaths, there had been no previous CPS involvement with the child's family. Although 61% of child abuse deaths were considered to be preventable, much of the responsibility for preventing these deaths rests with community members (eg, relatives, neighbors) who were aware of the abuse but failed to report the family to CPS. The CFRTs, who had received training in the proper completion of death certificates, reported that the cause of death was incorrect on 13% of all death certificates and in 16 cases, the CFRTs disagreed with the medical examiner on the manner of death (eg, natural, accidental, undetermined). Because CFRTs have access to additional information that may not have been available to the physician who completes a child's death certificate, CFRTs may be able to more accurately determine the cause and manner of death than the physician who completed the death certificate. Arizona's child death rate is above the national average (82.16/100 000), but the ACFRP determined that many of these deaths could have been prevented by using known prevention strategies (eg, child safety restraints, pool fencing). Most child mortality data are based on death certificate information that often is incorrect and cannot be used to assess preventability. Although most states have child fatality review programs that review suspected child abuse deaths, <3% of all preventable deaths in Arizona were attributable to child abuse. If all child deaths in the United States were reviewed from a prevention/needs assessment perspective, targeted and data-driven recommendations for prevention could be developed for each community, and potentially 38% of all child deaths that occur after the first month of life could be prevented. The ACFRP is an excellent example of a statewide system with a public health focus. To assist other states in developing similar programs, national support is needed. The establishment of a public health focused federally funded national program would provide us with the opportunity to standardize data collection among states and better utilize this data at a national level.


Molecular and cellular roles of PI31 (PSMF1) protein in regulation of proteasome function.
We investigated molecular features and cellular roles of PI31 (PSMF1) on regulation of proteasome function. PI31 has a C-terminal HbYX (where Hb is a hydrophobic amino acid, Y is tyrosine, and X is any amino acid) motif characteristic of several proteasome activators. Peptides corresponding to the PI31 C terminus also bind to and activate the 20 S proteasome in an HbYX-dependent manner, but intact PI31protein inhibits in vitro 20 S activity. Binding to and inhibition of the proteasome by PI31 are conferred by the HbYX-containing proline-rich C-terminal domain but do not require HbYX residues. Thus, multiple regions of PI31 bind independently to the proteasome and collectively determine effects on activity. PI31 blocks the ATP-dependent in vitro assembly of 26 S proteasome from 20 S proteasome and PA700 subcomplexes but has no effect on in vitro activity of the intact 26 S proteasome. To determine the physiologic significance of these in vitro effects, we assessed multiple aspects of cellular proteasome content and function after altering PI31 levels. We detected no change in overall cellular proteasome content or function when PI31 levels were either increased by moderate ectopic overexpression or decreased by RNA interference (RNAi). We also failed to identify a role of PI31 ADP-ribosylation as a mechanism for regulation of overall 26 S proteasome content and function, as recently proposed. Thus, despite its in vitro effects on various proteasome activities and its structural relationship to established proteasome regulators, cellular roles and mechanisms of PI31 in regulation of proteasome function remain unclear and require future definition.


Safety, tumor trafficking and immunogenicity of chimeric antigen receptor (CAR)-T cells specific for TAG-72 in colorectal cancer.
T cells engineered to express chimeric antigen receptors (CARs) have established efficacy in the treatment of B-cell malignancies, but their relevance in solid tumors remains undefined. Here we report results of the first human trials of CAR-T cells in the treatment of solid tumors performed in the 1990s. Patients with metastatic colorectal cancer (CRC) were treated in two phase 1 trials with first-generation retroviral transduced CAR-T cells targeting tumor-associated glycoprotein (TAG)-72 and including a CD3-zeta intracellular signaling domain (CART72 cells). In trial C-9701 and C-9702, CART72 cells were administered in escalating doses up to 1010 total cells; in trial C-9701 CART72 cells were administered by intravenous infusion. In trial C-9702, CART72 cells were administered via direct hepatic artery infusion in patients with colorectal liver metastases. In both trials, a brief course of interferon-alpha (IFN-α) was given with each CART72 infusion to upregulate expression of TAG-72. Fourteen patients were enrolled in C-9701 and nine in C-9702. CART72 manufacturing success rate was 100% with an average transduction efficiency of 38%. Ten patients were treated in CC-9701 and 6 in CC-9702. Symptoms consistent with low-grade, cytokine release syndrome were observed in both trials without clear evidence of on target/off tumor toxicity. Detectable, but mostly short-term (≤14 weeks), persistence of CART72 cells was observed in blood; one patient had CART72 cells detectable at 48 weeks. Trafficking to tumor tissues was confirmed in a tumor biopsy from one of three patients. A subset of patients had 111Indium-labeled CART72 cells injected, and trafficking could be detected to liver, but T cells appeared largely excluded from large metastatic deposits. Tumor biomarkers carcinoembryonic antigen (CEA) and TAG-72 were measured in serum; there was a precipitous decline of TAG-72, but not CEA, in some patients due to induction of an interfering antibody to the TAG-72 binding domain of humanized CC49, reflecting an anti-CAR immune response. No radiologic tumor responses were observed. These findings demonstrate the relative safety of CART72 cells. The limited persistence supports the incorporation of co-stimulatory domains in the CAR design and the use of fully human CAR constructs to mitigate immunogenicity.


Trading scalpels for sheaths: Catheter-based treatment of vascular injury can be effectively performed by acute care surgeons trained in endovascular techniques.
The skill set of the acute care surgeon can be expanded by formal training. We report the first series of traumatic vascular injury (TVI) treated by acute care surgeons trained in endovascular techniques (ACSTEV). We retrospectively reviewed patients admitted to our trauma center with TVI over 5 months who survived for more than 24 hours and had catheter diagnosis and/or therapy by ACSTEV. Demographics, admission data, and outcomes were reviewed. Follow-up ranged from 0 day to 150 days. Most patients were male (63%) and sustained blunt mechanism (91%). Mean (SD) age was 48.2 (21.9) years, and mean (SD) Injury Severity Score was 32.1 (11.8). Mean (SD) admission systolic blood pressure, heart rate, Glasgow Coma Scale (GCS) score were 126.12 (30.4) mm Hg, 101.21 (28.2) beats per minute, and 10.8 (4.73), respectively. Forty-six patients underwent 48 endovascular procedures for TVI: 32 angiograms and 16 venograms were obtained. Two pelvic angiograms and one aortic arch angiogram were negative and required no treatment. One superficial femoral artery arteriogram showed minor luminal defects requiring anticoagulation only. Pseudoaneurysms were found in 17 vessels, vessel truncation in 4, active extravasation in 5, stenosis in 1, and dissection with thrombus in 1. Four patients had resuscitative endovascular balloon occlusion of the aorta performed before catheter intervention for pelvic hemorrhage. Procedures included aortic repair (4), pelvic embolization (13), splenic embolization (5), lumbar artery embolization (1), bronchial artery embolization (1), profunda artery embolization (1), common carotid artery stent (1), celiac artery stent (1), inferior vena cava filter placement (14) and retrieval (2), and pharmacomechanical thrombolysis (1). Treatment material included coils (12), Gelfoam (4), and nitinol plugs (3). No procedural or device-related complications occurred. Mortality was 14.7% unrelated to any endovascular procedure. One patient had repeat coil embolization of a pelvic pseudoaneurysm on postoperative Day 7. ACSTEV can safely treat TVI with good success. We performed nearly 10 procedures per month underscoring the role of the ACSTEV for training and care of TVI in a high-volume trauma center. Therapeutic study, level V.


Spatial enhancer clustering and regulation of enhancer-proximal genes by cohesin.
In addition to mediating sister chromatid cohesion during the cell cycle, the cohesin complex associates with CTCF and with active gene regulatory elements to form long-range interactions between its binding sites. Genome-wide chromosome conformation capture had shown that cohesin's main role in interphase genome organization is in mediating interactions within architectural chromosome compartments, rather than specifying compartments per se. However, it remains unclear how cohesin-mediated interactions contribute to the regulation of gene expression. We have found that the binding of CTCF and cohesin is highly enriched at enhancers and in particular at enhancer arrays or "super-enhancers" in mouse thymocytes. Using local and global chromosome conformation capture, we demonstrate that enhancer elements associate not just in linear sequence, but also in 3D, and that spatial enhancer clustering is facilitated by cohesin. The conditional deletion of cohesin from noncycling thymocytes preserved enhancer position, H3K27ac, H4K4me1, and enhancer transcription, but weakened interactions between enhancers. Interestingly, ∼ 50% of deregulated genes reside in the vicinity of enhancer elements, suggesting that cohesin regulates gene expression through spatial clustering of enhancer elements. We propose a model for cohesin-dependent gene regulation in which spatial clustering of enhancer elements acts as a unified mechanism for both enhancer-promoter "connections" and "insulation."


Heteroaromatic monothiocarboxylic acids from Pseudomonas spp.
Pyridine derivatives substituted with monothiocarboxylic acid groups are the unique metabolites of certain Pseudomonas species. Pyridine-2,6-di-(monothiocarboxylic acid) 1a was found during a screening program for antibiotically active bacterial metabolites due to its ability to complex Fe3+. The structure of this complex, its redox behavior and the biogenesis of the ligand molecule were studied in detail. This lead to the discovery of a new class of natural products, viz. acylsulfenic acid derivatives. Interest in la was revived shortly when complexes with other metals were studied as models for sulfur-containing enzymes. It could also be shown that a quinoline monothiocarboxylic acid derivative acted as an alternative siderophore for Pseudomonas fluorescens. But a real renaissance was observed only when the role of la in the degradation of CCl4 by Pseudomonas stutzeri became evident.


Identification of N-terminal receptor activity-modifying protein residues important for calcitonin gene-related peptide, adrenomedullin, and amylin receptor function.
Calcitonin-family receptors comprise calcitonin receptor-like receptor (CL) or calcitonin receptor and receptor activity-modifying protein (RAMP) pairings. Calcitonin gene-related peptide (CGRP) receptors are CL/RAMP1, whereas adrenomedullin (AM) receptors are CL/RAMP2 (AM1 receptor) or CL/RAMP3 (AM2 receptor). Amylin (Amy) receptors are RAMP hetero-oligomers with the calcitonin receptor (AMY1, AMY2, and AMY3, respectively). How RAMPs change G protein-coupled receptor pharmacology is not fully understood. We exploited sequence differences between RAMP1 and RAMP3 to identify individual residues capable of altering receptor pharmacology. Alignment of human RAMPs revealed eight residues that are conserved in RAMP2 and RAMP3 but are different in RAMP1. We hypothesized that residues in RAMP2 and RAMP3, but not RAMP1, are responsible for making CL/RAMP2 and CL/RAMP3 AM receptors. Using site-directed mutagenesis, we introduced individual RAMP3 residues into RAMP1 and vice versa in these eight positions. Mutant or wild-type RAMPs were transfected into Cos7 cells with CL or the insert-negative form of the calcitonin receptor [CT(a)]. Agonist-stimulated cAMP production and cell-surface expression of constructs were measured. Position 74 in RAMP1 and RAMP3 was critical for determining AM potency and affinity, and Phe93 in RAMP1 was an important contributor to alphaCGRP potency at CGRP receptors. Mutant RAMP/CT(a) receptor complexes displayed different phenotypes. It is noteworthy that RAMP1 S103N and W74E mutations led to enhanced rAmy potency, probably related to increased cell-surface expression of these complexes. This differs from the effect on CL-based receptors where expression was unchanged. Targeted substitution has emphasized the importance of position 74 in RAMP1/RAMP3 as a key determinant of AM pharmacology.


Female brain size affects the assessment of male attractiveness during mate choice.
Mate choice decisions are central in sexual selection theory aimed to understand how sexual traits evolve and their role in evolutionary diversification. We test the hypothesis that brain size and cognitive ability are important for accurate assessment of partner quality and that variation in brain size and cognitive ability underlies variation in mate choice. We compared sexual preference in guppy female lines selected for divergence in relative brain size, which we have previously shown to have substantial differences in cognitive ability. In a dichotomous choice test, large-brained and wild-type females showed strong preference for males with color traits that predict attractiveness in this species. In contrast, small-brained females showed no preference for males with these traits. In-depth analysis of optomotor response to color cues and gene expression of key opsins in the eye revealed that the observed differences were not due to differences in visual perception of color, indicating that differences in the ability to process indicators of attractiveness are responsible. We thus provide the first experimental support that individual variation in brain size affects mate choice decisions and conclude that differences in cognitive ability may be an important underlying mechanism behind variation in female mate choice.


Taxa as types: Buffon, Cuvier and Lamarck.
From a Darwinian point of view, taxonomic groups are understood as historical entities that arise at an evolutionary moment and that can always disappear. But these groups were also understood by many naturalists as natural kinds; in other words, as permanent, ahistorical types. I will explore some of the forms that this typological thought took, showing that this typological perspective neither depends on theological beliefs, nor obeys the adoption of an ontology that might contradict natural science. Thus I shall analyze Buffon's understanding of species and the ways in which Cuvier and Lamarck understood the higher taxonomic orders.


[Gliomas and BRCA genes mutations: fortuitous association or imputability?].
BRCA is a tumor suppressor gene implicated in the major mechanisms of cellular stability in every type of cell. Its mutations are described in numerous cancers, mainly breast and ovarian in women. It was also found an increase of lifetime risk of pancreas, colon, prostate cancer or lymphoma in men carriers. We report the cases of two female patients aged 40 and 58-years-old female patients and one 35-years-old male patient, with brain or medullar gliomas, carriers of a germline mutation of BRCA gene. Those gliomas were particularly aggressive and were not responding to the standard treatment, with chemo and radiotherapy. The very unusual characteristics in location and evolutive profile of these central nervous system tumors raise the question of a genetical underlying mechanism, maybe linked to the BRCA gene mutation that carry these patients. In addition, a non-fortuitous association between germline mutation of BRCA and occurrence of a glioma can be evoked according to the embryological, epidemiological and biomolecular findings noted in the literature. Other clinical and experimental studies are necessary to precise the physiopathological link existing between BRCA mutations and the occurrence of a glioma; this could have therapeutical and clinical implications in the future.


Three-dimensional cone beam computed tomography analysis of temporomandibular joint response to the Twin-block functional appliance.
To propose a three-dimensional (3D) method for evaluating temporomandibular joint (TMJ) changes during Twin-block treatment. Seventeen patients with Class II division 1 malocclusion treated using Twin-block and nine untreated patients with a similar malocclusion were included in this research. We collected their cone beam computed tomography (CBCT) data from before and 8 months after treatment. Segmentations were constructed using ITK-SNAP. Condylar volume and superficial area were measured using 3D Slicer. The 3D landmarks were identified on CBCT images by using Dolphin software to assess the condylar positional relationship. 3D models of the mandible and glenoid fossa of the patients were constructed and registered via voxel-based superimposition using 3D Slicer. Thereafter, skeletal changes could be visualized using 3DMeshMetric in any direction of the superimposition on a color-coded map. All the superimpositions were measured using the same scale on the distance color-coded map, in which red color represents overgrowth and blue color represents resorption. Significant differences were observed in condylar volume, superficial area, and condylar position in both groups after 8 months. Compared with the control group (CG), the Twin-block group exhibited more obvious condyle-fossa modifications and joint positional changes. Moreover, on the color-coded map, more obvious condyle-fossa modifications could be observed in the posterior and superior directions in the Twin-block group than in the CG. We successfully established a 3D method for measuring and evaluating TMJ changes caused by Twin-block treatment. The treatment produced a larger condylar size and caused condylar positional changes.


Multidisciplinary care and units for uveitis in the internal medicine departments in Spain: Survey of the Systemic Autoimmune Diseases Group.
To identify the multidisciplinary uveitis units in which internal medicine departments participate in collaboration with ophthalmology departments in Spain. We conducted a multicentre, observational cross-sectional study that collected information using a structured survey sent by email to 1015 partners of the Systemic Autoimmune Diseases Workgroup of the Spanish Society of Internal Medicine (GEAS-SEMI) from the 1st to the 31st of March 2017. We identified a total of 21 support units/consultations for the ophthalmology departments. Seventeen (81%) of the units were specific internal medicine-ophthalmology consultations, and 5 (24%) units had been created in the past 5 years. A median of 460 patients were assessed per unit by the end of the year. This study shows, for the first time in Spain, the important and close collaboration between ophthalmologists and internists, especially in highly specialised national reference institutions.


Long-term survival with stereotactic radiotherapy for imaging-diagnosed pituitary tumors in dogs.
Published studies on the use of stereotactic radiotherapy for dogs with pituitary tumors are limited. This retrospective observational study describes results of stereotactic radiotherapy for 45 dogs with imaging-diagnosed pituitary tumors. All dogs were treated at a single hospital during the period of December 2009-2015. The stereotactic radiotherapy was delivered in one 15 Gray (Gy) fraction or in three 8 Gy fractions. At the time of analysis, 41 dogs were deceased. Four were alive and censored from all survival analyses; one dog received 8 Gy every other day and was removed from protocol analyses. The median overall survival from first treatment was 311 days (95% confidence interval 226-410 days [range 1-2134 days]). Thirty-two dogs received 15 Gy (median overall survival 311 days; 95% confidence interval [range 221-427 days]), and 12 received 24 Gy on three consecutive days (median overall survival 245 days, 95% confidence interval [range 2-626 days]). Twenty-nine dogs had hyperadrenocorticism (median overall survival 245 days), while 16 had nonfunctional masses (median overall survival 626 days). Clinical improvement was reported in 37/45 cases. Presumptive signs of acute adverse effects within 4 months of stereotactic radiotherapy were noted in 10/45, and most had improvement spontaneously or with steroids. Late effects versus tumor progression were not discernable, but posttreatment blindness (2), hypernatremia (2), and progressive neurological signs (31) were reported. There was no statistical difference in median overall survival for different protocols. Patients with nonfunctional masses had longer median overall survival than those with hyperadrenocorticism (P = 0.0003). Survival outcomes with stereotactic radiotherapy were shorter than those previously reported with definitive radiation, especially for dogs with hyperadrenocorticism.


Porous nitrogen-doped carbon nanofibers assembled with nickel nanoparticles for lithium-sulfur batteries.
High-efficiency cathodes in lithium-sulfur (Li-S) batteries play an important role in the pursuit of high electrochemical performance. However, a number of Li-S batteries currently reported suffer from severe drawbacks such as the insulating nature of sulfur, sluggish redox kinetics, and the shuttle effect of intermediate polysulfides. To overcome these challenges, herein, carbonyl group functionalized porous carbon nanofibers assembled with nickel (Ni/PCNFO) are proposed to serve as an efficient sulfur host in Li-S batteries. Such a Ni/PCNFO-S composite cathode exhibits outstanding electrochemical performances, which are attributed to three factors: (1) the large inner space of the PCNF can afford a high S content and accommodate the volume expansion; (2) high electrical conductivity is provided by the carbon nanofiber skeleton and the electrocatalytically active Ni species embedded in the PCNF significantly facilitate the redox kinetics of the S species; and (3) the carbonyl group anchored on the Ni/PCNF can effectively suppress the polysulfide effect via strong chemical affinity/adsorption with polysulfides. With these advantageous features, the Li-S batteries based on Ni/PCNFO-S cathodes exhibit a high specific capacity (1320 mA h g-1), excellent rate capability (780 mA h g-1), and long cycling stability (910 mA h g-1 after 500 cycles at 0.2C).


A spatial model for nutrient mitigation potential of blue mussel farms in the western Baltic Sea.
Worldwide, coastal and marine policies are increasingly aiming for environmental protection, and eutrophication is a global challenge, particularly impairing near-coastal marine water bodies. In this context, mussel mitigation aquaculture is currently considered an effective tool to extract nutrients from such water bodies. Mussel mitigation farming using longline systems with loops of collector material is a well-developed technology and considered promising in the western Baltic Sea. Besides several spatially limited field studies, a suitable spatial model for site-specific implementation is still lacking. In this study, we present a modular spatial model, consisting of a spatial and temporal habitat factor model (Module 1), blue mussel growth model (Module 2), mussel farm model (Module 3), and an avoidance of food limitation model (Module 4). The modules integrate data from in situ monitoring, mussel growth experiments, and eco-physiological modelling for the western Baltic Sea, to estimate spatially explicit nutrient reduction potentials. The model is flexible with respect to farm setups and harvest times and considers natural variability, model uncertainty, and required hydrodynamics. Modelling results proved valid at all scales and modules, and point out key areas for efficient mussel mitigation farms in Danish, German and Swedish areas. Modelled long-term mean mitigation potentials for harvest in November reach up to 0.88 tN/ha and 0.05 tP/ha for a farm setup using 2 m depth-range of the water column and 3.0 tN/ha and 0.17 tP/ha using up to 8 m, respectively. For Danish water bodies, we demonstrate that in efficient areas, mitigation farms (18.8 ha, 90 km collector substrate in loops with 2 m depth-range) required <3.6% of the space to extract the target nitrogen loads for good ecological status. The developed approach could prove valuable for implementing environmental policies in aquatic systems, e.g. in situ nutrient mitigation, aquaculture spatial planning, and habitat suitability mapping.


Ultrasound-assisted extraction of antioxidants in Misai Kucing (Orthosiphon stamineus).
Ultrasound-assisted extraction (UAE) with ethanol was used to extract the compounds responsible for the antioxidant activities of Misai Kucing (Orthosiphon stamineus). Response surface methodology (RSM) was used to optimize four independent variables: ethanol concentration (%), amplitude (%), duty cycle (W/s) and extraction time (min). Antioxidant compounds were determined by total phenolic content and total flavonoid content to be 1.4 g gallic acid equivalent/100 g DW and 45 g catechin equivalent/100 g DW, respectively. Antioxidant activities were evaluated using the 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+) radical scavenging capacity assay and the 2,2-diphenyl-1-picrylhydrazyl (DPPH•) radical scavenging capacity assay to be 1,961.3 and 2,423.3 µmol Trolox Equivalent Antioxidant Capacity (TEAC)/100 g DW, respectively. Based on the optimal conditions, experimental values were reported to be close to the predicted value by RSM modeling (p>0.05), indicating the suitability of UAE for extracting the antioxidants of Misai Kucing. Rosmarinic acid, kaempferol-rutinoside and sinesetine were identified by high performance liquid chromatography-mass spectrometry.


Intermittent compressive force promotes osteogenic differentiation in human periodontal ligament cells by regulating the transforming growth factor-β pathway.
Mechanical force regulates periodontal ligament cell (PDL) behavior. However, different force types lead to distinct PDL responses. Here, we report that pretreatment with an intermittent compressive force (ICF), but not a continuous compressive force (CCF), promoted human PDL (hPDL) osteogenic differentiation as determined by osteogenic marker gene expression and mineral deposition in vitro. ICF-induced osterix (OSX) expression was inhibited by cycloheximide and monensin. Although CCF and ICF significantly increased extracellular adenosine triphosphate (ATP) levels, pretreatment with exogenous ATP did not affect hPDL osteogenic differentiation. Gene-expression profiling of hPDLs subjected to CCF or ICF revealed that extracellular matrix (ECM)-receptor interaction, focal adhesion, and transforming growth factor beta (TGF-β) signaling pathway genes were commonly upregulated, while calcium signaling pathway genes were downregulated in both CCF- and ICF-treated hPDLs. The TGFB1 mRNA level was significantly increased, while those of TGFB2 and TGFB3 were decreased by ICF treatment. In contrast, CCF did not modify TGFB1 expression. Inhibiting TGF-β receptor type I or adding a TGF-β1 neutralizing antibody attenuated the ICF-induced OSX expression. Exogenous TGF-β1 pretreatment promoted hPDL osteogenic marker gene expression and mineral deposition. Additionally, pretreatment with ICF in the presence of TGF-β receptor type I inhibitor attenuated the ICF-induced mineralization. In conclusion, this study reveals the effects of ICF on osteogenic differentiation in hPDLs and implicates TGF-β signaling as one of its regulatory mechanisms.


Nutrition and physical activity during and after cancer treatment: an American Cancer Society guide for informed choices.
Cancer survivors are often highly motivated to seek information about food choices, physical activity, dietary supplement use, and complementary nutritional therapies to improve their treatment outcomes, quality of life, and survival. To address these concerns, the American Cancer Society (ACS) convened a group of experts in nutrition, physical activity, and cancer to evaluate the scientific evidence and best clinical practices related to optimal nutrition and physical activity after the diagnosis of cancer. This report summarizes their findings and is intended to present health care providers with the best possible information on which to help cancer survivors and their families make informed choices related to nutrition and physical activity. The report discusses nutrition and physical activity issues during the phases of cancer treatment and recovery, living after recovery from treatment, and living with advanced cancer; selected nutritional and physical activity issues such as body weight, food choices, and complementary and alternative nutritional options; and selected issues related to breast, colorectal, lung, prostate, head and neck, and upper gastrointestinal cancers. In addition, handouts containing commonly asked questions and answers and a resource list are provided for survivors and families. Tables that grade the scientific evidence for benefit versus harm related to nutrition and physical activity for breast, colorectal, lung, and prostate cancers are also included for this growing body of knowledge to provide guidance for informed decision making and to identify areas for future research.


Calcium signalling and gene expression.
A wide variety of compounds acting as extracellular signals cause changes in the free cytosolic Ca2+ concentration. These factors include hormones, growth factors, neurotransmitters, but also nutrient and metabolic activators. Ca2+ signalling is caused by mobilization of Ca2+ from internal stores and by well controlled and timed Ca2+ influx from the extracellular space. Ca2+ signals address Ca2+ dependent enzymes, most importantly Ca2+ sensitive protein kinases and phosphatases. The profound influence of Ca2+ signalling on gene expression has been recognized a long time ago. As Ca2+ signals are short-lived when compared to alterations in differentiated gene expression, it is generally considered that genes coding for short-lived transcription factors (i.e. fos, jun) are the immediate target of Ca2+ signalling. Transcription of these immediate early genes (IEG) can be activated without the need for protein synthesis. Ca2+ signalling affects differentiated gene expression via changes in the absolute and relative abundance of IEG products, which in turn control the expression of differentiated genes. Ca2+ signals can stimulate both transcriptional initiation as well as transcriptional elongation. Initiation of transcription is stimulated by the Ca2+ dependent phosphorylation of binding proteins addressing two response elements in the promoter of IEGs: the cAMP response element, CRE, and the serum response element, SRE. Distinct protein kinases are involved in either case. We study the elongation of transcripts of the IEG c-fos beyond the first intron which is favoured by Ca2+ signals, involving mechanisms which still are poorly understood. We can show that intron sequences contribute to the control of elongation by Ca2+, and that there is a strong interrelation between the transcription control by the promoter and by the intron.


Role of systems pharmacology in understanding drug adverse events.
Systems pharmacology involves the application of systems biology approaches, combining large-scale experimental studies with computational analyses, to the study of drugs, drug targets, and drug effects. Many of these initial studies have focused on identifying new drug targets, new uses of known drugs, and systems-level properties of existing drugs. This review focuses on systems pharmacology studies that aim to better understand drug side effects and adverse events. By studying the drugs in the context of cellular networks, these studies provide insights into adverse events caused by off-targets of drugs as well as adverse events-mediated complex network responses. This allows rapid identification of biomarkers for side effect susceptibility. In this way, systems pharmacology will lead to not only newer and more effective therapies, but safer medications with fewer side effects.


Caring for patients in a malpractice crisis: physician satisfaction and quality of care.
The rhetoric of malpractice reform is at fever pitch, but political advocacy does not necessarily reflect grassroots opinion. To determine whether the ongoing liability crisis has greatly reduced physicians' professional satisfaction, we surveyed specialist physicians in Pennsylvania. We found widespread discontent among physicians practicing in high-liability environments, which seems to be compounded by other financial and administrative pressures. Opinion alone should not determine public policy, but physicians' perceptions matter for two reasons. First, perceptions influence behavior with respect to practice environment and clinical decision making. Second, perceptions influence the physician-patient relationship and the interpersonal quality of care.


Use of communication technologies to cost-effectively increase the availability of interpretation services in healthcare settings.
Poor patient-provider communication due to limited English proficiency (LEP) costs healthcare providers and payers through lower patient use of preventive care, misdiagnosis, increased testing, poor patient compliance, and increased hospital and emergency room admissions. Scarcity of bilingual healthcare professionals and prohibitive interpretation costs hinder full implementation of language service despite federal and state laws requiring their provision. We review recent published literature and unpublished data documenting the use of telephonic and video interpretation methodologies to improve healthcare communication with LEP persons. For example, a cooperative of nine California public hospitals and their associated community clinics, psychiatric facilities, skilled nursing facilities, and public health departments have implemented shared video interpretation services with video/voice-over Internet Protocol call center technology that automatically routes requests for interpretation in 15 languages to a pool of 30 full-time interpreters and 4 trained bilingual staff. For organizations seeking to initiate or expand their language services, the Internet provides access to translated documents, promising practices, step-by-step guides, planning tools, and research briefs. Such recent technological advances make provision of language services-to respond to federal and state mandates and improve access and quality of care to LEP persons-more feasible than is widely believed. Increased government and foundation support, and collaboration among provider organizations themselves can catalyze these efforts.


Microbiota, oral microbiome, and pancreatic cancer.
Only 30% of patients with a diagnosis of pancreatic cancer survive 1 year after the diagnosis. Progress in understanding the causes of pancreatic cancer has been made, including solidifying the associations with obesity and diabetes, and a proportion of cases should be preventable through lifestyle modifications. Unfortunately, identifying reliable biomarkers of early pancreatic cancer has been extremely challenging, and no effective screening modality is currently available for this devastating form of cancer. Recent data suggest that the microbiota may play a role in the disease process, but many questions remain. Future studies focusing on the human microbiome, both etiologically and as a marker of disease susceptibility, should shed light on how to better tackle prevention, early detection, and treatment of this highly fatal disease.


Implications for pathogenesis of patterns of injury in small- and medium-sized-vessel vasculitis.
The different pathologic features of different types of necrotizing vasculitis indicate that there are different pathogenic mechanisms causing the injury. The pathogenic mechanisms for medium-sized-vessel vasculitis are most effective at causing injury in arteries and are not effective at causing injury in smaller vessels. The predilection of medium-sized-vessel vasculitis for bifurcations may relate to the increased expression of adhesion molecules and increased numbers of intimal macrophages at these sites. The preferential involvement of small vessels by small-vessel vasculitis may relate to the requirement for close apposition between leukocytes and endothelial cells for the pathogenic mechanisms to be operational. The pathology of the necrotizing vasculitis of Kawasaki disease is most consistent with a primary role for monocytes/macrophages and T lymphocytes in the acute injury. The pathology of the necrotizing vasculitis of polyarteritis nodosa and small-vessel vasculitis, including ANCA-vasculitis, is most consistent with a primary role for neutrophils and monocytes in the acute injury.


[The role of chest X-rays in the diagnosis of thoracic aortic aneurysms].
To evaluate the role of routine chest radiographs in the diagnosis of thoracic aortic aneurysms (TAA). An electronic full-text search was performed in our radiological information system for all patients who underwent chest radiograph under standard conditions between 1998 and 2000 and who had suspected widening or aneurysm of the thoracic aorta as a diagnosis. Computed tomography (CT) of the thorax was used as the gold standard and had to be performed within a period of 30 days. Two independent and blinded observers evaluated different morphologic and morphometric parameters in the diagnosis and correlated the results with those of CT. 28 patients were included in the present trial. With almost perfect interobserver correlation (r = 0.95) both investigated morphometric parameters correlated well (r = 0.85 and 0.83) with the diameter of the aorta as evaluated with CT. While a low subjective over-all probability for TAA had a negative predictive value of 100 %, we found that, despite an almost perfect interobserver variability (Kappa > 0.8), none of the investigated morphologic parameters (discrepancy between the ascending and descending aorta, displacement of the trachea to the right and caudal displacement of the left main bronchus) was significantly correlated with the final diagnosis. The investigated morphometric parameters help to estimate the diameter of the aorta in the arch and in the descending section, but none of the morphologic criteria can be used for the diagnosis of TAA.


Early initiation of lipid-lowering therapy for acute coronary syndromes improves compliance with guideline recommendations: observations from the Orbofiban in Patients with Unstable Coronary Syndromes (OPUS-TIMI 16) trial.
Lipid-lowering is effective in the prevention of cardiovascular morbidity and mortality in patients with coronary artery disease, but effective strategies for improving the implementation of these therapies are needed. In the 10,288 patients in the OPUS-TIMI 16 trial, patients were stratified by use of lipid-lowering therapy during index hospitalization and were compared for use of lipid-lowering therapy at follow-up as well as for clinical outcomes. Lipid-lowering therapy was used in 38% of patients during the index hospitalization, of which 94% were statins. At 10 months, 88% of patients who were discharged on lipid-lowering medications remained on these drugs. Conversely, only 34% of patients not discharged on lipid-lowering medications were receiving them at 10 months. Forty-one percent of patients with prior history of hyperlipidemia requiring treatment were not discharged on lipid-lowering therapy, and of these, only 51% were subsequently started on a lipid-lowering medication as an outpatient despite clear indications. Patients treated as inpatients with lipid-lowering therapy had a lower mortality rate at 10 months adjusted by propensity analysis (3.1% vs 5.1%, P < .0001) than patients not treated with lipid-lowering therapy. In patients with acute coronary syndromes, the initiation of lipid-lowering therapy in the inpatient setting increases the rate of its subsequent use at 10 months, making this an important method of ensuring appropriate secondary prevention.


Is p53 a protein that predicts the response to chemotherapy in node negative breast cancer?
The role of p53 in modulating apoptosis has suggested that it may affect efficacy of anti cancer agents. For this reason, we have evaluated p53 alterations in 282 consecutive patients with infiltrating node-negative breast cancer who underwent primary surgery and were randomized either to CMF (Cyclophosphamide 400 mg/m2, Fluorouracil 400 mg/m2, and Methotrexate 40 mg/m2) or control arm (no adjuvant therapy) from 1980 to 1989. p53 alterations were analyzed by immunohistochemistry using DO7 MoAb, revealed by immunoperoxidase technique, and quantitated in term of percentage of positive cells. We observed a positive staining in 24% of the tumors. Among them, 10% had a positive staining in more than 75% of the cells. There was a highly significant association between the proportion of positive cells and histologic grade of the infiltrating ductal carcinomas (p<0.004). However, there was no association with age, tumor size, hormone receptor content, or vascular embolism. There was a trend but no significant relationship between positive staining and overall survival either in each arm of the trial or in the overall population. Interestingly, we observed a higher relative risk of local relapse after conservative therapy in the boosted area in the group of mutated p53 (RR=4.41; p<0.0005). We conclude that, in this node-negative breast tumor population, alteration of p53 cannot predict the response to the chemotherapy. However, it may represent a useful marker of risk of local relapse and of radio resistance.


Surface bound radicals, char yield and particulate size from the burning of tobacco cigarette.
Tobacco smoke is a toxic gas-phase cocktail consisting of a broad range of organics, and free radical intermediates. The formation of smoke from a burning cigarette depends on a series of mechanisms, including generation of products by pyrolysis and combustion, aerosol formation, and physical mass transfer processes. The current study simulates the deposition of particulate matter on the human lung surface by trapping the tobacco smoke particulates in situ on silica gel. To mimic this phenomenon, the cigarette was smoked directly on siliga gel. The surface morphology of smoke condensate trapped on silica gel, and pure silica gel (control) was investigated using a scanning electron microscope (SEM). Electron paramagnetic resonance (EPR) was used to explore the presence of free radicals on the particulate matter trapped on silica. Standard procedures for cigarette smoking (ISO 3402:1999) were adopted. The char yields of tobacco cigarette in the temperature range 200-700 °C was also investigated in an inert atmosphere using a quartz reactor. SEM images showed the surface morphology of pure silica gel was smooth while silica gel on which cigarette smoke was smoked on contained particulates of various sizes. Generally, the particulate size of cigarette smoke adsorbed on silica was found to be 2.47 ± 0.0043 µm (~PM2.5). Electron paramagnetic resonance (EPR) results showed a g-value of 2.0037 typically that of a carbon-centred radical. It is therefore evident from this investigation that cigarette smoke contains surface bound radicals considered harmful to the health of cigarette smokers. The particulate size of tobacco smoke (PM2.5) can impact severely on the lives of the cigarette smoking community because of its near ultrafine nature. This significantly small particulate size in cigarette smoke can be inhaled deeper into the lungs thus causing serious cell injury and possible tumour growth in addition to other grave diseases. Graphical abstract Cigarette smoking and analytical techniques employed in this study.


cSPARCOM: Multi-detector reconstruction by confocal super-resolution correlation microscopy.
Image scanning microscopy (ISM), an upgraded successor of the ubiquitous confocal microscope, facilitates up to two-fold improvement in lateral resolution, and has become an indispensable element in the toolbox of the bio-imaging community. Recently, super-resolution optical fluctuation image scanning microscopy (SOFISM) integrated the analysis of intensity-fluctuations information into the basic ISM architecture, to enhance its resolving power. Both of these techniques typically rely on pixel-reassignment as a fundamental processing step, in which the parallax of different detector elements to the sample is compensated by laterally shifting the point spread function (PSF). Here, we propose an alternative analysis approach, based on the recent high-performing sparsity-based super-resolution correlation microscopy (SPARCOM) method. Through measurements of DNA origami nano-rulers and fixed cells labeled with organic dye, we experimentally show that confocal SPARCOM (cSPARCOM), which circumvents pixel-reassignment altogether, provides enhanced resolution compared to pixel-reassigned based analysis. Thus, cSPARCOM further promotes the effectiveness of ISM, and particularly that of correlation based ISM implementations such as SOFISM, where the PSF deviates significantly from spatial invariance.


[The content of beta-endorphin, met-enkephalin and ACTH in the blood plasma during electric and drug anesthesia].
Beta-endorphin, metenkephaline, and ACTH were radioimmunoassayed in the peripheral blood plasma of 4 groups of patients subjected to microsurgery by different techniques. In 3 groups electropulsed exposure of the CNS was a component of general combined ataralgesia. In group 1 (13 patients) two Lenar devices were employed, in group 2 (5 patients) Skat-202 device, in group 3 (7 patients) Elean device. Group 4 (17 patients) were controls administered drugs according to the same protocols as in the rest three groups. Anesthesia was considered adequate in all the groups. The hypoalgesic effect of electroexposure was the most expressed in group 1: fentanyl was not injected in 53% cases, and in 47% its dose, 0.74 microgram/kg/h, was 2.25 times lower than in controls (1.58 micrograms/kg/hm p < 0.05); the drug doses in groups 2 and 3 (0.82 and 0.8 microgram/kg/h) were 1.9 and 2 times lower than in controls, respectively, p < 0.05. The levels of ACTH and opioid peptides were measured at 6 stages: 1) several days before surgery; 2) after premedication, 10-15 min after the patient was brought into the operation room; 3) before discontinuing nitrogen oxide; 4) after discontinuing nitrogen oxide; 5) when the patient regained consciousness after the end of surgery; and 6) after extubation of the trachea. During surgery under electromedicamentous anesthesia the level of ACTH in the plasma was the same as initially or comparable to the level of this hormone at the same stages in the control group. The content of opioid peptides changed at stressogenic stages of anesthesia and surgery. In patients administered drug anesthesia beta-endorphin levels were shifted, in those operated on under electromedicamentous anesthesia the metenkephaline compound of the opiate system was altered. Activation of various components of endogenous opiate system in electromedicamentous and drug anesthesia may be due to differences in the mechanisms of this types of anesthesia at the given level of the antinociceptive system of the organism. No stable reaction of endorphin level on the electroexposure could be detected. There was no evident relationship between changes in the levels of opioid peptides and the decrease of fentanyl consumption. Besides, it is possible that the analgesic effect of electroexposure is mediated not only by the opioid, but by other mechanisms of endogenous antinociceptive system as well.


CMM-Net: Contextual multi-scale multi-level network for efficient biomedical image segmentation.
Medical image segmentation of tissue abnormalities, key organs, or blood vascular system is of great significance for any computerized diagnostic system. However, automatic segmentation in medical image analysis is a challenging task since it requires sophisticated knowledge of the target organ anatomy. This paper develops an end-to-end deep learning segmentation method called Contextual Multi-Scale Multi-Level Network (CMM-Net). The main idea is to fuse the global contextual features of multiple spatial scales at every contracting convolutional network level in the U-Net. Also, we re-exploit the dilated convolution module that enables an expansion of the receptive field with different rates depending on the size of feature maps throughout the networks. In addition, an augmented testing scheme referred to as Inversion Recovery (IR) which uses logical "OR" and "AND" operators is developed. The proposed segmentation network is evaluated on three medical imaging datasets, namely ISIC 2017 for skin lesions segmentation from dermoscopy images, DRIVE for retinal blood vessels segmentation from fundus images, and BraTS 2018 for brain gliomas segmentation from MR scans. The experimental results showed superior state-of-the-art performance with overall dice similarity coefficients of 85.78%, 80.27%, and 88.96% on the segmentation of skin lesions, retinal blood vessels, and brain tumors, respectively. The proposed CMM-Net is inherently general and could be efficiently applied as a robust tool for various medical image segmentations.


Planning and decision making about the future care of older group home residents and transition to residential aged care.
Planning for future care after the death of parental caregivers and adapting disability support systems to achieve the best possible quality of life for people with intellectual disability as they age have been important issues for more than two decades. This study examined perceptions held by family members, group home staff and organisational managers about the future of older residents and the decisions made that a move to residential aged care was necessary. Grounded Dimensional Analysis was used to guide data collection and analysis by an interdisciplinary research team. Three sets of interviews over a period of 18 months were conducted with a family member, house supervisor and the programme manager for each of seventeen older group home residents in Victoria. For the eight people for whom it was decided a move was necessary and the six who eventually moved focussed questions were asked about the decision-making process. While plans for lifelong accommodation in a group home proved unfounded, key person succession plans were effective. However, decisions to move to a residential aged care facility where necessary were made in haste and seen as a fait accompli by involved family members. Although family members take seriously their mandate to oversee well-being of their older relative, they have little knowledge about their rights or avenues to safeguard untimely or inappropriate decisions being made by professionals.


Mixing of diphtheria, tetanus, and acellular pertussis (DTaP) vaccines in a population of children in managed care.
The Advisory Committee on Immunization Practices recommends administering diphtheria, tetanus and acellular pertussis (DTaP) vaccines to children at 2, 4, 6, 15-18 months, and 4-6 y of age; preferably with the same-brand vaccine for the whole series. We estimated age-appropriate DTaP dose completion and the proportion of children receiving a "mixed" DTaP vaccination series (ie, including DTaP vaccines from ≥ 2 brands) across the 3 milestones. Commercially-insured children born between 01/01/2003 and 04/30/2011 were identified from United States health insurance claims data and assigned to ≥ 1 of 3 study cohorts based on the duration of continuous health plan enrollment: 1) birth to <8 months; 2) birth to <20 months; 3) birth to <7 years. Dose completion and brand mixing of the first 3, first 4 or all 5 doses were measured in the respective cohorts. Administered DTaP vaccinations were identified in claims data and classified by brand (based on vaccine components and manufacturer). The analysis included children who received ≥ 2 DTaP vaccinations and had known brand information for all doses. Age-appropriate dose completion was 77% with 3 doses (<8 months cohort), 71% with 4 doses (<20 months cohort), and 85% with 5 doses (<7 years cohort). Mixed DTaP series were received by 4.7% (95% confidence interval [CI]: 4.6%-4.7%) in the <8 months cohort, 29.0% (95% CI: 28.6%-29.4%) in the <20 months cohort, and 39.0% (95% CI: 34.5, 43.6) in the <7 years cohort. DTaP mixing was just 4.7% for the first 3 doses but subsequently increased with the number of administered doses.


Analysis of genetic diversity of Chinese dairy goats via microsatellite markers.
In this study, 15 polymorphic microsatellite markers were used to analyze the genetic structure and phylogenetic relationships of 6 dairy goat breeds in China, including 4 native developed breeds and 2 introduced breeds. The results showed that a total of 172 alleles were detected in 347 samples of the dairy goat breeds included in this study. The mean number of effective alleles per locus was 4.92. Except for BMS0812, all of the remaining microsatellite loci were highly polymorphic (polymorphism information content [PIC] > 0.5). The analysis of genetic diversity parameters, including the number of effective alleles, PIC, and heterozygosity, revealed that the native developed dairy goat breeds in China harbored a rich genetic diversity. However, these breeds showed a low breeding degree and a high population intermix degree, with a certain degree of inbreeding and within-subpopulation inbreeding coefficient ( > 0). The analysis of population genetic differentiation and phylogenetic tree topologies showed a moderate state of genetic differentiation among subpopulations of native developed breed dairy goats in China (0.05 < gene fixation coefficient [] < 0.15). The native developed breeds shared a common ancestor, namely, the Saanen dairy goat, originating from Europe. The results showed that there was a close genetic relationship between Wendeng and Laoshan dairy goats while the Guanzhong dairy goat and the Xinong Saanen dairy goat were also found to have a close genetic relationship, which were both in agreement with the formation history and geographical distribution of the breeds. This study revealed that adopting genetic management strategies, such as expanding pedigree source and strengthening multi-trait selection, is useful in maintaining the genetic diversity of native developed breeds and improving the population uniformity of dairy goats.


Can we use fixed ambient air monitors to estimate population long-term exposure to air pollutants? The case of spatial variability in the Genotox ER study.
Associations between average total personal exposures to PM2.5, PM10, and NO2 and concomitant outdoor concentrations were assessed within the framework of the Genotox ER study. It was carried out in four French metropolitan areas (Grenoble, Paris, Rouen, and Strasbourg) with the participation, in each site, of 60-90 nonsmoking volunteers composed of two groups of equal size (adults and children) who carried the personal Harvard Chempass multipollutant sampler during 48 h along two different seasons ("hot" and "cold"). In each center, volunteers were selected so as to live (home and work/school) in three different urban sectors contrasted in terms of air pollution (one highly exposed to traffic emissions, one influenced by local industrial sources, and a background urban environment). In parallel to personal exposure measurements, a fixed ambient air monitoring station surveyed the same pollutants in each local sector. A linear regression model was accommodated where the dependent pollutant-specific variable was the difference, for each subject, between the average ambient air concentrations over 48 h and the personal exposure over the same period. The explanatory variables were the metropolitan areas, the three urban sectors, season, and age group. While average exposures to particles were underestimated by outdoor monitors, in almost all cities, seasons, and age groups, differences were lower for NO2 and, in general, in the other direction. Relationships between average total personal exposures and ambient air levels varied across metropolitan areas and local urban sectors. These results suggest that using ambient air concentrations to assess average exposure of populations, in epidemiological studies of long-term effects or in a risk assessment setting, calls for some caution. Comparison of personal exposures to PM or NO2 with ambient air levels is inherently disturbed by indoor sources and activities patterns. Discrepancies between measurement devices and local and regional sources of pollution may also strongly influence how the ambient air concentrations relate to population exposure. Much attention should be given to the selection of the most appropriate monitoring sites according to the study objectives.


Assessment of muscle activity and fatigue during laparoscopic surgery.
Cumulative musculoskeletal stress during operative procedures can contribute to the development of chronic musculoskeletal injury among surgeons. This is a concern in laparoscopic specialties where trainees may incur greater risk by learning poor operative posture or technique early in training. This study conducted an initial investigation of the physical stress encountered during the conduct of foregut laparoscopic surgery. Subjects were divided into two groups based on their surgical experience level, high experience (HE), consisting of two attending surgeons, and low experience (LE), consisting of two fellow surgeons and a surgical chief resident. Nine distinct foregut laparoscopic procedures were observed for data collection within these groups. Electromyographic (EMG) activity was collected at the bilateral neck, shoulders, biceps, triceps, and lower back for each procedure. Physical workload was measured using percent reference voluntary contractions (%RVC) for each surgeon's muscle activities. Fatigue development was assessed using the median frequency of EMG data between two consecutive cases. Subjects completed a NASA-TLX survey when surgery concluded. LE surgeons experienced higher levels of %RVC in the lower back muscles compared to HE surgeons. LE fatigue level was also higher than HE surgeons across most muscle groups. A decrease in median frequency in six of the ten muscle groups after performing two consecutive cases, the largest decrements being in the biceps and triceps indicated fatigue development across consecutive cases for both surgeon groups. Surgeons developed fatigue in consecutive cases while performing minimally invasive surgery (MIS). HE surgeons demonstrated a lower overall physical workload while also demonstrating different patterns in muscle work. The findings from this study can be used to inform further ergonomic studies and the data from this study can be used to develop surgical training programs focused on the importance of surgeon ergonomics and minimizing occupational injury risk.


Vegan Diet Advice Might Benefit Liver Enzymes in Nonalcoholic Fatty Liver Disease: an Open Observational Pilot Study.
The Western diet is rich in saturated fats, refined sugars and meat consistent with a high-energy load and secondary risk of increased metabolic diseases including nonalcoholic fatty liver disease (NAFLD). However, no data are available on potential benefit of vegan diets in NAFLD and/or nonalcoholic steatohepatitis (NASH). We aimed to study prospectively the effect of a vegan diet, excluding all animal products on liver chemistry in a group of consecutive NAFLD patients. This was a prospective, pilot study run on 40 consecutive patients affected by NAFLD. Eight subjects refused to join the study for poor diet palatability, leaving 32 patients (19 males, mean age 50 years), with abnormal measures of liver function who agreed to adhere to a vegan diet for six months. The caloric intake was tailored by the dietitian to obtain a weight loss ≥5% of body weight in overweight patients [body-mass index (BMI) ≥25] and ranged from 1500 Kcal to 1800 Kcal. Patients were contacted monthly by phone to reinforce diet and lifestyle advice and were seen at the gastrointestinal clinic when doubtful about diet advice. At six-month follow-up, 6 subjects did not attend the clinic leaving only 26 patients for data analysis. Initial anthropometric values were mean weight 78 kg (range 52-95), mean body mass index (BMI) 26.8 Kg/m2 (range 20.3-31.2). Liver function tests showed mean ALT value 99 U/L (SD±45), mean AST value 54 U/L (SD±44), mean GGT value 160 U/L (SD±122), pre-treatment. After six months mean body weight was 73 Kg (range 52-87), mean BMI was 25.2 Kg/m2 (range 20.3-29.7) (p<0.001 compared to baseline for both parameters). Liver enzymes improved to a mean of ALT value 36 U/L (SD±21), AST value 27 U/L (SD±10) and GGT value 55 U/L (SD±57), respectively (p<0.001 compared to baseline for all enzymes). Normalization of liver function tests as a whole was observed in 20/26 patients (76.9%). A loss of ≥ 5% of body weight was observed in 12 patients (46.1%), but it did not correlate with the normalization of liver function tests (p=0.5). Our data provide preliminary evidence of improved liver enzymes in NAFLD patients with a strict vegan diet and although our study sample is limited, decreased body weight did not seem critical to the outcome.


Hydrophobic Nanoparticles Modify the Thermal Release Behavior of Liposomes.
Understanding the effect of embedded nanoparticles on the characteristics and behavior of lipid bilayers is critical to the development of lipid-nanoparticle assemblies (LNAs) for biomedical applications. In this work we investigate the effect of hydrophobic nanoparticle size and concentration on liposomal thermal release behavior. Decorated LNAs (D-LNAs) were formed by embedding 2 nm (GNP2) and 4 nm (GNP4) dodecanethiol-capped gold nanoparticles into DPPC liposomes at lipid to nanoparticle ratios (L:N) of 25,000:1, 10,000:1, and 5,000:1. D-LNA structure was investigated by cryogenic transmission electron microscopy, and lipid bilayer permeability and phase behavior were investigated based on the leakage of a model drug, carboxyfluorescein, and by differential scanning calorimetry, respectively. The presence of bilayer nanoparticles caused changes in the lipid bilayer release and phase behavior compared to pure lipid controls at very low nanoparticle to bilayer volume fractions (0.3%-4.6%). Arrhenius plots of the thermal leakage show that GNP2 led to greater increases in the leakage energy barrier compared to GNP4, consistent with GNP4 causing greater bilayer disruption due to their size relative to the bilayer thickness. Embedding hydrophobic nanoparticles as permeability modifiers is a unique approach to controlling liposomal leakage based on nanoparticle size and concentration.


tRNA gene clusters at the 3' end of rRNA operons are specific to virulent subgroups of Streptococcus agalactiae strains, as demonstrated by molecular differential analysis at the population level.
The aim of this work was to characterize a 2.4 kb randomly amplified polymorphic DNA (RAPD) fragment described as a marker for a phylogenetic group of Streptococcus agalactiae strains significantly associated with neonatal meningitis. This fragment was analysed by cloning and sequencing, and showed that two types of tRNA gene cluster flank the 3' end of the rRNA operons in S. agalactiae strains. Both types of tRNA gene cluster act as markers for phylogenetic subgroups of strains within the species. One type could be used to distinguish two of the three virulent intraspecies subgroups to which most of the S. agalactiae strains able to invade the central nervous system of neonates belong. This raises the possibility that there is a link between these tRNA genes and the virulence of the bacterium. Based on this analysis, PCR primers were designed to determine whether S. agalactiae strains are likely to belong to lineages of organisms in which most of the highly virulent strains isolated from cerebrospinal fluid cluster. In addition, this work demonstrated that RAPD can be used to detect novel particularities within intraspecies variants of pathogens.


Crystal structure of the cold-adapted haloalkane dehalogenase DpcA from Psychrobacter cryohalolentis K5.
Haloalkane dehalogenases (HLDs) convert halogenated aliphatic pollutants to less toxic compounds by a hydrolytic mechanism. Owing to their broad substrate specificity and high enantioselectivity, haloalkane dehalogenases can function as biosensors to detect toxic compounds in the environment or can be used for the production of optically pure compounds. Here, the structural analysis of the haloalkane dehalogenase DpcA isolated from the psychrophilic bacterium Psychrobacter cryohalolentis K5 is presented at the atomic resolution of 1.05 Å. This enzyme exhibits a low temperature optimum, making it attractive for environmental applications such as biosensing at the subsurface environment, where the temperature typically does not exceed 25°C. The structure revealed that DpcA possesses the shortest access tunnel and one of the most widely open main tunnels among structural homologs of the HLD-I subfamily. Comparative analysis revealed major differences in the region of the α4 helix of the cap domain, which is one of the key determinants of the anatomy of the tunnels. The crystal structure of DpcA will contribute to better understanding of the structure-function relationships of cold-adapted enzymes.


Chagas disease.
Chagas disease is a chronic, systemic, parasitic infection caused by the protozoan Trypanosoma cruzi, and was discovered in 1909. The disease affects about 8 million people in Latin America, of whom 30-40% either have or will develop cardiomyopathy, digestive megasyndromes, or both. In the past three decades, the control and management of Chagas disease has undergone several improvements. Large-scale vector control programmes and screening of blood donors have reduced disease incidence and prevalence. Although more effective trypanocidal drugs are needed, treatment with benznidazole (or nifurtimox) is reasonably safe and effective, and is now recommended for a widened range of patients. Improved models for risk stratification are available, and certain guided treatments could halt or reverse disease progression. By contrast, some challenges remain: Chagas disease is becoming an emerging health problem in non-endemic areas because of growing population movements; early detection and treatment of asymptomatic individuals are underused; and the potential benefits of novel therapies (eg, implantable cardioverter defibrillators) need assessment in prospective randomised trials.


Water-Soluble Polymeric Carbon Nitride Colloidal Nanoparticles for Highly Selective Quasi-Homogeneous Photocatalysis.
Heptazine-based polymeric carbon nitrides (PCN) are promising photocatalysts for light-driven redox transformations. However, their activity is hampered by low surface area resulting in low concentration of accessible active sites. Herein, we report a bottom-up preparation of PCN nanoparticles with a narrow size distribution (ca. 10±3 nm), which are fully soluble in water showing no gelation or precipitation over several months. They allow photocatalysis to be carried out under quasi-homogeneous conditions. The superior performance of water-soluble PCN, compared to conventional solid PCN, is shown in photocatalytic H2 O2 production via reduction of oxygen accompanied by highly selective photooxidation of 4-methoxybenzyl alcohol and benzyl alcohol or lignocellulose-derived feedstock (ethanol, glycerol, glucose). The dissolved photocatalyst can be easily recovered and re-dissolved by simple modulation of the ionic strength of the medium, without any loss of activity and selectivity.


Analysis of the hemispheric asymmetry using fractal dimension of a skeletonized cerebral surface.
We investigated hemispheric asymmetry using the fractal dimension (FD) of the skeletonized cerebral surface. Sixty-two T1-weighted magnetic resonance imaging volumes from normal Korean adults were used. The skeletonization of binary volume data, which corresponded to the union of the gray matter and cerebrospinal flow classified by fuzzy clustering, was performed slice by slice in the sagittal direction, and then skeletonized slices were integrated into the three-dimensional (3-D) hemisphere. Finally, the FD of the 3-D skeletonized cerebral surface was calculated using the box-counting method. We measured the FD of the skeletonized cerebral surface and the volumes of intracranial gray matter and white matter for the whole hemispheres and obtained the hemispheric asymmetries of each measurement. The FD, the gray matter, and the white matter volumes for the whole hemispheres decreased in the old group. The asymmetry of the FD revealed a significant right-greater-than-left asymmetry showed rightward, but did not change according to age and gender. None of the intracranial gray matter or white matter volumes showed any significant asymmetric changes. It could be said that the FD of the skeletonized cerebral surface is a novel measure of cerebral asymmetry.


Isolation and characterization of Tri3, a gene encoding 15-O-acetyltransferase from Fusarium sporotrichioides.
An acetyltransferase gene (Tri3) was isolated from Fusarium sporotrichioides by complementation of a previously identified Tri3- mutant and shown to be closely linked to three other trichothecene biosynthetic pathway genes. Comparison of the Tri3 sequence with its cDNA revealed the presence of four introns. The Tri3 cDNA contains a 1,539-bp open reading frame that encodes a protein with a molecular mass of 57,418 Da. Regulation of Tri3 transcription in liquid cultures appeared identical to that of other trichothecene pathway genes. Disruption of the Tri3 gene resulted in the accumulation of deacetylated calonectrins rather than T-2 toxin. The results of whole-cell feeding experiments with Tri3- strains suggested that 15-O-acetylation is blocked. Cell-free feeding experiments confirmed that Tri3- strains are able to acetylate a trichothecene C-3 hydroxyl group but are unable to acetylate a trichothecene C-15 hydroxyl group. Our results show that Tri3 encodes an acetyltransferase that converts 15-decalonectrin to calonectrin.


Acidic microenvironment created by osteoclasts causes bone pain associated with tumor colonization.
Bone pain is one of the most common complications in cancer patients with bone metastases. Previous findings that inhibitors of osteoclastic bone resorption such as bisphosphonates (BPs) reduce bone pain suggest a critical role of osteoclasts. Osteoclasts destroy bone by secreting protons, thereby making adjacent microenvironment acidic. Because acidosis is a well-known cause of pain, it is plausible that an osteoclasts-created acidic microenvironment may cause bone pain associated with cancer colonization in bone. To test this notion, we studied an animal model in which inoculation of MRMT-1 rat breast cancer cells into the tibiae in female rats induced hyperalgesia. Radiographic and histological analyses demonstrated that MRMT-1 cells caused aggressive bone destruction with an increased number of osteoclasts. Behavioral analyses showed that rats exhibited hyperalgesia in the tumor-inoculated legs. The BP zoledronic acid (ZOL) significantly reduced the hyperalgesia. In addition, immunohistochemical examinations revealed that c-Fos expression in the ipsilateral spinal cord neurons was increased. ZOL decreased these c-Fos-positive neurons. To investigate the role of acidosis, mRNA expression of acid-sensing receptors including acid-sensing channels (ASICs) and transient receptor potential channel-vanilloid subfamily member 1 (TRPV1) in the dorsal root ganglions (DRGs) was determined. The expression of ASIC1a and ASIC1b was increased in the ipsilateral DRGs, whereas the ASIC3 and TRPV1 expression was not changed. Of note, ZOL reduced the expression of ASIC1a and ASIC1b. In conclusion, our data suggest that an acidic microenvironment created by osteoclasts, at least in part, contributes to the induction of hyperalgesia through upregulating ASICs expression.


Suicide Prevention in an Emergency Department Population: The ED-SAFE Study.
Suicide is a leading cause of deaths in the United States. Although the emergency department (ED) is an opportune setting for initiating suicide prevention efforts, ED-initiated suicide prevention interventions remain underdeveloped. To determine whether an ED-initiated intervention reduces subsequent suicidal behavior. This multicenter study of 8 EDs in the United States enrolled adults with a recent suicide attempt or ideation and was composed of 3 sequential phases: (1) a treatment as usual (TAU) phase from August 2010 to December 2011, (2) a universal screening (screening) phase from September 2011 to December 2012, and (3) a universal screening plus intervention (intervention) phase from July 2012 to November 2013. Screening consisted of universal suicide risk screening. The intervention phase consisted of universal screening plus an intervention, which included secondary suicide risk screening by the ED physician, discharge resources, and post-ED telephone calls focused on reducing suicide risk. The primary outcome was suicide attempts (nonfatal and fatal) over the 52-week follow-up period. The proportion and total number of attempts were analyzed. A total of 1376 participants were recruited, including 769 females (55.9%) with a median (interquartile range) age of 37 (26-47) years. A total of 288 participants (20.9%) made at least 1 suicide attempt, and there were 548 total suicide attempts among participants. There were no significant differences in risk reduction between the TAU and screening phases (23% vs 22%, respectively). However, compared with the TAU phase, patients in the intervention phase showed a 5% absolute reduction in suicide attempt risk (23% vs 18%), with a relative risk reduction of 20%. Participants in the intervention phase had 30% fewer total suicide attempts than participants in the TAU phase. Negative binomial regression analysis indicated that the participants in the intervention phase had significantly fewer total suicide attempts than participants in the TAU phase (incidence rate ratio, 0.72; 95% CI, 0.52-1.00; P = .05) but no differences between the TAU and screening phases (incidence rate ratio, 1.00; 95% CI, 0.71-1.41; P = .99). Among at-risk patients in the ED, a combination of brief interventions administered both during and after the ED visit decreased post-ED suicidal behavior.


Uric acid attenuates trophoblast invasion and integration into endothelial cell monolayers.
Hyperuricemia develops as early as 10 wk of gestation in women who later develop preeclampsia. At this time the invasive trophoblast cells are actively remodeling the uterine spiral arterioles, integrating into and finally replacing the vascular endothelial lining. In the nonpregnant population uric acid has several pathogenic effects on vascular endothelium. We therefore sought to examine the effects of uric acid (0-7 mg/dl) on trophoblast cell invasion through an extracellular matrix using an in vitro Matrigel invasion assay. We also assessed trophoblast integration into a uterine microvascular endothelial cell monolayer in a trophoblast-endothelial cell coculture model. Additionally, we addressed the importance of redox signaling and trophoblast-induced endothelial cell apoptosis. Uric acid elicited a concentration-dependent attenuation of trophoblast invasion and integration into a uterine microvascular endothelial cell monolayer. The attenuated trophoblast integration appeared to be the result of reduced trophoblast-induced endothelial cell apoptosis, likely through the intracellular antioxidant actions of uric acid. In a test of relevance, pooled serum (5% vol/vol) from preeclamptic women attenuated the ability of trophoblast cells to integrate into the endothelial cell monolayers compared with pooled serum from healthy pregnant controls, and this response was partially rescued when endogenous uric acid was previously removed with uricase. Taken together these data support the hypothesis that elevations in circulating uric acid in preeclamptic women contribute to the pathogenesis of the disorder, in part, through attenuation of normal trophoblast invasion and spiral artery vascular remodeling.


Minireview: What is Known about SUMOylation Among NR4A Family Members?
NR4A receptors, including NUR77 (NR4A1), NURR1 (NR4A2) and NOR-1 (NR4A3), form a family of nuclear receptors that act as transcription factors to regulate many physiological and pathological processes such as cell cycle and apoptosis, lipid metabolism, inflammation, carcinogenesis, vascular and neuronal functions. In the absence of known endogenous ligand modulating their physiological functions, the NR4A family remains a class of orphan receptors. However, several post-translational modifications (PTMs), including SUMOylation, have been shown to regulate the expression and/or activity of these receptors. Addition of Small Ubiquitin-like MOdifier (SUMO) proteins is a dynamic and reversible enzymatic process that regulates multiple essential functions of proteins, including nuclear receptors. This review aims at summarizing what is known about the impact of SUMOylation on NR4A family member transcriptional activities and physiological functions.


Factors associated with serological cure and the serofast state of HIV-negative patients with primary, secondary, latent, and tertiary syphilis.
Some syphilis patients remain in a serologically active state after the recommended therapy. We currently know too little about the characteristics of this serological response. We conducted a cohort study using the clinical database from Zhongshan Hospital, Medical College of Xiamen. In total, 1,327 HIV-negative patients with primary, secondary, latent, and tertiary syphilis were enrolled. Bivariate and multivariate analyses were utilised to identify factors associated with a serological cure and serofast state in syphilis patients one year after therapy. Chi-square tests were used to determine the differences in the serological cure rate across different therapy time points. One year after the recommended therapy, 870 patients achieved a serological cure, and 457 patients (34.4%) remained in the serofast state. The serological cure rate increased only within the first 6 months. The bivariate analysis indicated that male or younger patients had a higher likelihood of a serological cure than female or older patients. Having a baseline titre ≤ 1∶2 or ≥ 1∶64 was associated with an increased likelihood of a serological cure. The serological cure rate decreased for the different disease stages in the order of primary, secondary, latent, and tertiary syphilis. A distinction should be drawn between early and late syphilis. The multivariate analysis indicated that a serological cure was significantly associated with the disease phase, gender, age, and baseline rapid plasma reagin (RPR) titre. The serofast state is common in clinical work. After one year of the recommended therapy, quite a few syphilis patients remained RPR positive. The primary endpoint of the study indicated that disease phase, gender, age and baseline RPR titre were crucial factors associated with a serological cure.


CBS9106-induced CRM1 degradation is mediated by cullin ring ligase activity and the neddylation pathway.
Chromosome region maintenance 1 (CRM1) mediates the nuclear export of proteins and mRNAs, and is overexpressed in various cancers. Recent studies have also reported that CRM1 protein expression is a negative prognostic factor in patients with cancer. Therefore, CRM1 is considered a potential target for anticancer therapy. Our previous study demonstrated that CBS9106, a synthetic small-molecular inhibitor of CRM1, decreases CRM1 protein through proteasomal degradation without affecting CRM1 mRNA levels. However, the mechanism by which CRM1 is degraded is not well understood. Here, we demonstrate a novel signaling pathway that plays an important role in CBS9106-induced CRM1 degradation. We found that MLN4924, a selective inhibitor of NEDD8-activating enzyme (NAE), effectively inhibits cullin neddylation and attenuates CBS9106-induced CRM1 degradation in a time- and dose-dependent manner. MLN4924 also attenuated CBS9106-induced nuclear accumulation of Ran-binding protein 1 (RanBP1), cell growth inhibition, and apoptosis. Furthermore, RNAi-mediated knockdown of neddylation pathway proteins (NEDD8 and UBA3) or cullin ring ligase (CRL) component protein (Rbx1) attenuated CRM1 protein degradation and G1 phase cell-cycle arrest by CBS9106. Knockdown of CSN5 or CAND1 also partially inhibited CBS9106-induced CRM1 degradation. These findings demonstrate that CBS9106-induced CRM1 degradation is conferred by CRL activity involving the neddylation pathway, and that this response to CBS9106 leads to cell growth inhibition and apoptosis.


Gene expression changes during caste-specific neuronal development in the damp-wood termite Hodotermopsis sjostedti.
One of the key characters of social insects is the division of labor, in which different tasks are allocated to various castes. In termites, one of the representative groups of social insects, morphological differences as well as behavioral differences can be recognized among castes. However, very little is known about the neuronal and molecular bases of caste differentiation and caste-specific behavior. In almost all termite species, soldiers play defensive roles in their colonies, and their morphology and behavior are largely different from workers (or pseudergates). Therefore, we predicted that some genes linked to defensive behavior and/or those required for neuronal changes are differentially expressed between workers and soldiers, or during the soldier differentiation, respectively. Using the brain and suboesophageal ganglion (SOG) of the damp-wood termite Hodotermopsis sjostedti, we first screened genes specifically expressed in soldiers or during soldier differentiation by the differential display method, followed by quantitative real-time polymerase chain reaction. No distinctive differences in expression patterns were detected between pseudergates and soldiers. In the course of soldier differentiation, however, five genes were found to be up-regulated in brain and/or SOG: 14-3-3 epsilon, fibrillin2, beta-tubulin, ciboulot, and a hypothetical protein containing a SAP motif. Some of these genes are thought to be associated with cytoskeletal structure or motor-associated proteins in neuronal tissues. The identified five genes could be involved in soldier-specific neuronal modifications, resulting in defensive behaviors in termite soldiers. The temporal expression patterns of these genes were consistent with the neuronal changes during soldier differentiation, suggesting that molecular machineries, in which the identified factors would participate, play important roles in behavioral differentiation of termite soldiers.


Optical second harmonic generation from nanostructured graphene: a full wave approach.
Optical second harmonic generation (SHG) from nanostructured graphene has been studied in the framework of classical electromagnetism using a surface integral equation method. Single disks and dimers are considered, demonstrating that the nonlinear conversion is enhanced when a localized surface plasmon resonance is excited at either the fundamental or second harmonic frequency. The proposed approach, beyond the electric dipole approximation used in the quantum description, reveals that SHG from graphene nanostructures with centrosymmetric shapes is possible when retardation effects and the excitation of high plasmonic modes at the second harmonic frequency are taken into account. Several SHG effects similar to those arising in metallic nanostructures, such as the silencing of the nonlinear emission and the design of double resonant nanostructures, are also reported. Finally, it is shown that the SHG from graphene disk dimers is very sensitive to a relative vertical displacement of the disks, opening new possibilities for the design of nonlinear plasmonic nanorulers.