The N-end rule pathway counteracts cell death by destroying proapoptotic protein fragments.
In the course of apoptosis, activated caspases cleave ∼500 to ∼1,000 different proteins in a mammalian cell. The dynamics of apoptosis involve a number of previously identified, caspase-generated proapoptotic protein fragments, defined as those that increase the probability of apoptosis. In contrast to activated caspases, which can be counteracted by inhibitor of apoptosis proteins, there is little understanding of antiapoptotic responses to proapoptotic protein fragments. One possibility is the regulation of proapoptotic fragments through their selective degradation. The previously identified proapoptotic fragments Cys-RIPK1, Cys-TRAF1, Asp-BRCA1, Leu-LIMK1, Tyr-NEDD9, Arg-BID, Asp-BCL(XL), Arg-BIM(EL), Asp-EPHA4, and Tyr-MET bear destabilizing N-terminal residues. Tellingly, the destabilizing nature (but not necessarily the actual identity) of N-terminal residues of proapoptotic fragments was invariably conserved in evolution. Here, we show that these proapoptotic fragments are short-lived substrates of the Arg/N-end rule pathway. Metabolic stabilization of at least one such fragment, Cys-RIPK1, greatly augmented the activation of the apoptosis-inducing effector caspase-3. In agreement with this understanding, even a partial ablation of the Arg/N-end rule pathway in two specific N-end rule mutants is shown to sensitize cells to apoptosis. We also found that caspases can inactivate components of the Arg/N-end rule pathway, suggesting a mutual suppression between this pathway and proapoptotic signaling. Together, these results identify a mechanistically specific and functionally broad antiapoptotic role of the Arg/N-end rule pathway. In conjunction with other apoptosis-suppressing circuits, the Arg/N-end rule pathway contributes to thresholds that prevent a transient or otherwise weak proapoptotic signal from reaching the point of commitment to apoptosis.


FAAH-/- mice display differential tolerance, dependence, and cannabinoid receptor adaptation after delta 9-tetrahydrocannabinol and anandamide administration.
Repeated administration of Delta(9)-tetrahydrocannabinol (THC), the primary psychoactive constituent of Cannabis sativa, induces profound tolerance that correlates with desensitization and downregulation of CB(1) cannabinoid receptors in the CNS. However, the consequences of repeated administration of the endocannabinoid N-arachidonoyl ethanolamine (anandamide, AEA) on cannabinoid receptor regulation are unclear because of its rapid metabolism by fatty acid amide hydrolase (FAAH). FAAH(-/-) mice dosed subchronically with equi-active maximally effective doses of AEA or THC displayed greater rightward shifts in THC dose-effect curves for antinociception, catalepsy, and hypothermia than in AEA dose-effect curves. Subchronic THC significantly attenuated agonist-stimulated [(35)S]GTP gamma S binding in brain and spinal cord, and reduced [(3)H]WIN55,212-2 binding in brain. Interestingly, AEA-treated FAAH(-/-) mice showed less CB(1) receptor downregulation and desensitization than THC-treated mice. Experiments examining tolerance and cross-tolerance indicated that the behavioral effects of THC, a low efficacy CB(1) receptor agonist, were more sensitive to receptor loss than those of AEA, a higher efficacy agonist, suggesting that the expression of tolerance was more affected by the intrinsic activity of the ligand at testing than during subchronic treatment. In addition, the CB(1) receptor antagonist, rimonabant, precipitated a markedly reduced magnitude of withdrawal in FAAH(-/-) mice treated subchronically with AEA compared with mice treated repeatedly with THC. The findings that repeated AEA administration produces lesser adaptive changes at the CB(1) receptor and has reduced dependence liability compared with THC suggest that pharmacotherapies targeting endocannabinoid catabolic enzymes are less likely to promote tolerance and dependence than direct acting CB(1) receptor agonists.


Membrane-Tethered Metalloproteinase Expressed by Vascular Smooth Muscle Cells Limits the Progression of Proliferative Atherosclerotic Lesions.
The MMP (matrix metalloproteinase) family plays diverse and critical roles in directing vascular wall remodeling in atherosclerosis. Unlike secreted-type MMPs, a member of the membrane-type MMP family, MT1-MMP (membrane-type 1 MMP; MMP14), mediates pericellular extracellular matrix degradation that is indispensable for maintaining physiological extracellular matrix homeostasis. However, given the premature mortality exhibited by MT1-MMP-null mice, the potential role of the proteinase in atherogenesis remains elusive. We sought to determine the effects of both MT1-MMP heterozygosity and tissue-specific gene targeting on atherogenesis in APOE (apolipoprotein E)-null mice. MT1-MMP heterozygosity in the APOE-null background (Mmp14+/-Apoe-/- ) significantly promoted atherogenesis relative to Mmp14+/+Apoe-/- mice. Furthermore, the tissue-specific deletion of MT1-MMP from vascular smooth muscle cells (VSMCs) in SM22α-Cre(+)Mmp14F/FApoe-/- (VSMC-knockout) mice likewise increased the severity of atherosclerotic lesions. Although VSMC-knockout mice also developed progressive atherosclerotic aneurysms in their iliac arteries, macrophage- and adipose-specific MT1-MMP-knockout mice did not display this sensitized phenotype. In VSMC-knockout mice, atherosclerotic lesions were populated by hyperproliferating VSMCs (smooth muscle actin- and Ki67-double-positive cells) that were characterized by a proinflammatory gene expression profile. Finally, MT1-MMP-null VSMCs cultured in a 3-dimensional spheroid model system designed to mimic in vivo-like cell-cell and cell-extracellular matrix interactions, likewise displayed markedly increased proliferative potential. MT1-MMP expressed by VSMCs plays a key role in limiting the progression of atherosclerosis in APOE-null mice by regulating proliferative responses and inhibiting the deterioration of VSMC function in atherogenic vascular walls.


Proteomics of beta2-microglobulin amyloid fibrils.
Knowledge on the chemical structure of beta2-microglobulin in natural amyloid fibrils is quite limited because of the difficulty in obtaining tissue samples suitable for biochemical studies. We have reviewed the available information on the chemical modifications and we present new data of beta2-microglobulin extracted from non-osteotendinous tissues. beta2-microglobulin can accumulate in these compartments after long-term haemodialysis but rarely forms amyloid deposits. We confirm that truncation at the N-terminus is an event specific to beta2-microglobulin derived from fibrils but is not observed in the beta2-microglobulin from plasma or from the insoluble non-fibrillar material deposited in the heart and spleen. We also confirm the partial deamidation of Asn 17 and Asn 42, as well as the oxidation of Met 99 in fibrillar beta2-microglobulin. Other previously reported chemical modifications cannot be excluded, but should involve less than 1-2% of the intact molecule.


[Translation of eukaryotic mRNA in a cap-independent mode].
All eukaryotic mRNA molecules have a cap structure at the 5' ends which plays a crucial role in the scanning model of their translation initiation. In an alternative way of translation, the active ribosome is formed in a cap-independent mode due to the presence of IRES, internal ribosome entry site, in the 5' untranslated region of certain mRNAs. This region folds into a distinct secondary and tertiary structure, which binds the 40S ribosomal subunit and some protein factors, and subsequently forms the initiation complex and the translationally active 80S ribosome. This enables the synthesis of specific proteins under the conditions when cap-dependent translation is inhibited or strongly reduced. The cap-independent mode of translation initiation concerns proteins that play very important roles during cell cycle, apoptosis, response to stress stimuli and cancer development.


Counseling for HIV testing.
Recommendations are presented for counseling individuals who wish to know if they have been infected by the human immunodeficiency virus (HIV). Pretest counseling includes explaining the sensitivity and meaning of HIV antibody tests and the limits of confidentiality, assessing the patient's potential strengths, vulnerabilities, coping capacity, and supportive resources, and mutually deciding if the test is advisable. Populations that have been identified as at risk for HIV infection are listed, and four case vignettes that illustrate the complexity of the decision to be tested are provided. Posttest counseling includes notification of test results, reducing the immediate distress of seropositive patients, educating about how HIV is and is not spread, explaining methods to prevent transmission, advising the patient about who should be told the test results, and arranging follow-up care.


Tissue-Specific Control of the Endocycle by the Anaphase Promoting Complex/Cyclosome Inhibitors UVI4 and DEL1.
The endocycle represents a modified mitotic cell cycle that in plants is often coupled to cell enlargement and differentiation. Endocycle onset is controlled by activity of the Anaphase Promoting Complex/Cyclosome (APC/C), a multisubunit E3 ubiquitin ligase targeting cell-cycle factors for destruction. CELL CYCLE SWITCH52 (CCS52) proteins represent rate-limiting activator subunits of the APC/C. In Arabidopsis (Arabidopsis thaliana), mutations in either CCS52A1 or CCS52A2 activators result in a delayed endocycle onset, whereas their overexpression triggers increased DNA ploidy levels. Here, the relative contribution of the APC/CCCS52A1 and APC/CCCS52A2 complexes to different developmental processes was studied through analysis of their negative regulators, being the ULTRAVIOLET-B-INSENSITIVE4 protein and the DP-E2F-Like1 transcriptional repressor, respectively. Our data illustrate cooperative activity of the APC/CCCS52A1 and APC/CCCS52A2 complexes during root and trichome development, but functional interdependency during leaf development. Furthermore, we found APC/CCCS52A1 activity to control CCS52A2 expression. We conclude that interdependency of CCS52A-controlled APC/C activity is controlled in a tissue-specific manner.


Metal speciation dynamics and bioavailability: Zn(II) and Cd(II) uptake by mussel (Mytilus edulis) and carp (Cyprinus carpio).
In the analysis of metal biouptake from complexing environments, both chemical speciation and biological uptake characteristics have to be taken into account. The commonly used free ion activity model is based on equilibrium speciation and implies that diffusion of the bioactive free metal toward the organism is not rate-limiting. In the presence of complexes, however, sufficiently labile species might contribute to the biouptake via preceding dissociation. Coupling of the ensuing diffusional mass transfer flux of metal with the biouptake flux of free metal, the supposedly bioactive species, shows under which conditions labile metal complexes can contribute to the uptake. The goal of the present paper is to apply this type of analysis to experimental data on metal uptake by mussel (Mytilus edulis) and carp (Cyprinus carpio) in complexing environments. These biosystems have fairly well-characterized uptake parameters, but the uptake fluxes cannot be fully explained by considering equilibrium speciation only. For Zn(II) uptake by mussel, evidence was found for diffusional limitation at low concentrations, whereas for Cd(III) uptake by carp, diffusion is not limiting at all. The analysis provides an example of how a more comprehensive treatment of complex systems can be applied to real experimental data.


Assessment of antibiotic prescriptions in pediatric care units in a regional university hospital.
The main objective was to assess the relevance of antibiotic prescriptions in the pediatric wards of a regional university hospital in France. Secondary objectives were to assess adequacy of the dose, administration frequency, administration route, treatment duration, adaptation to bacteriological results, and treatment reevaluation. We assessed antibiotic prescriptions in pediatric settings. We included all patients under 18 years of age hospitalized in a pediatric ward who received a computerized prescription for antibiotic treatment between June 1st and June 30th, 2018; 163 clinical cases for 157 patients were analyzed. Patients hospitalized in neonatology, pediatric intensive care unit, and onco-hematology wards were excluded. The rate of relevance was 71%. The rates of adequacy for the other criteria were 60% for the dose, 99% for the administration frequency, 98% for the administration route, 72% for treatment duration, 98% for treatment adaptation to microbiological results, and 100% for treatment revaluation. All criteria combined, the overall rate of adequacy was 28%. Effort should be made regarding doses and treatment durations. Areas for improvement have been suggested to the wards: standardized prescription protocols, pediatric prescription guide, training of residents, setting up of the operational team in pediatric wards and a second clinical evaluation.


Protective effect of dimethyl fumarate for the development of pressure ulcers after cutaneous ischemia-reperfusion injury.
Ischemia-reperfusion (I/R) is associated with various pathogenic conditions, and there has been increasing evidence that cutaneous I/R injury is associated with the pathogenesis of pressure ulcers (PUs), especially at the early stage presenting as non-blanchable erythema. Several studies demonstrated that oxidative stress is a key player in I/R injury, and the inhibition of oxidative stress may be capable of protecting tissue damage after I/R injury in various organs including skin. Dimethyl fumarate (DMF) approved by the Food and Drug Administration is Nrf2 activator, and recent studies revealed the antioxidative and anti-inflammatory effects of DMF on I/R injury in animal models. Our objective was to assess the effects of oral administration of DMF on the development of PUs after cutaneous I/R injury in mice. We found that DMF administration significantly decreased the size of PUs after cutaneous I/R. Cutaneous I/R-induced oxidative stress was also significantly inhibited by DMF in OKD48 mice, in which oxidative stress can be visually assessed. In addition, DMF treatment decreased hypoxic area, the numbers of apoptotic cells, and vascular loss in I/R area. DMF treatment suppressed the infiltration of MPO+ neutrophils and the production of proinflammatory cytokines in I/R site after cutaneous I/R injury. in vitro experiments, DMF treatment suppressed the production of reactive oxygen species in pericyte-like cells. These results suggest that DMF treatment might prevent the formation of PUs induced by cutaneous I/R injury via suppressing oxidative stress and subsequent inflammation. DMF treatment during the early phase of decubitus ulcers might protect against further progression.


Preoperative Axillary US in Early-Stage Breast Cancer: Potential to Prevent Unnecessary Axillary Lymph Node Dissection.
Purpose To evaluate the value of preoperative axillary ultrasonography (US) for preventing unnecessary axillary lymph node dissection after sentinel lymph node (SLN) biopsy in a large series of patients with early-stage breast cancers treated with both breast-conserving surgery and SLN biopsy. Materials and Methods From March 2009 to February 2013, 1802 patients who underwent breast-conserving surgery for clinical T1-2/N0 cancers and SLN biopsy with or without axillary lymph node dissection were included. Preoperative axillary US results and clinical-pathologic variables were compared according to the status of non-SLN metastasis. Multivariate logistic regression analysis was performed to find factors associated with non-SLN metastasis. Results Of 1802 patients, 397 (22.0%) underwent axillary lymph node dissection due to positive SLN biopsy and 76 (4.2%) had non-SLN metastasis at final histopathologic examination. Patients with non-SLN metastasis were younger and showed positive axilla at US and clinical T2 stage more frequently (P < .05). At multivariate analysis, positive axilla at US (P = .001), clinical T2 stage (P = .005), and lymphovascular invasion (P < .001) were significantly associated with non-SLN metastasis. Among 1284 patients with negative axilla at US and clinical T1 stage cancer, 1254 (97.7%) did not have non-SLN metastasis and 30 (2.3%) had non-SLN metastasis. Conclusion Preoperative axillary US results and clinical T stage are associated with the status of non-SLN metastasis in patients with early breast cancer. The results of this study suggest that preoperative axillary US can help select patients at minimal risk of non-SLN metastasis, for whom axillary lymph node dissection can be omitted. © RSNA, 2018 Online supplemental material is available for this article.


[Stapled hemorrhoidectomy--early experience in 30 patients].
We report the early results of 30 patients treated by stapled hemorrhoidectomy. Thirty patients with symptomatic grade 2-4 hemorrhoids were treated by stapled hemorrhoidectomy. The procedure was performed with a 33 mm diameter automatic hemorrhoidal circular stapler. The patients were prospectively evaluated for immediate and functional recovery, postoperative pain and subjective success of the treatment. The median age of the patients was 46.2 years, and the median duration of the symptoms was 27 months. The main symptom was bleeding (in 96% of the patients). The average operative time was 22 minutes. The operation was performed with spinal (63.3%) or general (36.7%) anesthesia. There was no mortality, urinary retention, incontinence, fecal urgency or persistent pain. One patient, who was under anticoagulant treatment, had postoperative bleeding, which required transfusion of 3 units of blood, and another patient was operated on because of perforation of the sigmoid colon. Most patients (twenty eight) had complete functional recovery and returned to their usual daily activities within 10.4 days. Postoperative pain and subjective success were evaluated by a 1 to 10 scale. The average pain score decreased from 5.8 on the first postoperative day to 2.4 on the 7th postoperative day. The average satisfactory score was 9.2. Stapled hemorrhoidectomy is an alternative to conventional surgical hemorrhoidectomy. The procedure seems to be associated with less postoperative pain and early recovery with a high satisfaction rate among patients.


Influence of cheese making process on STEC bacteriophage release.
Shiga toxin-producing Escherichia coli (STEC) are foodborne pathogens implicated in diseases including hemolytic uremic syndrome (HUS) and hemorrhagic colitis (HC). The main virulence factor are Shiga toxins; their production and secretion are by-products of the expression of late genes of prophages upon sub-lethal environmental stimuli exposure. Hence, the lysogenic prophage after a stress switch to lytic cycle spreading the Stx phages. In the present study, 35 STEC were screened for the presence and the ability to release Shiga toxin-encoding bacteriophages. Three bacterial strains showed signals of prophage presence both in plate and in PCR. Subsequently, these bacterial strains were subjected to stressors that simulate cheese manufacturing conditions: NaCl (1, 1.5 and 2% w/v), lactic acid (0.5, 1.5 and 3% v/v), anaerobic growth, pasteurization (72°C for 15 s), UV irradiation. The ability to release prophage was evaluated by Real Time qPCR. Induction of the prophages showed that the addition of NaCl at 1.5 and 2% significantly increased viral release compared to control. Conversely, the addition of lactic acid had a significant repressive effect. The other applied stressors had no significant effect in phage release according to the experimental conditions adopted.


Pediatric and Adult Patients Have Similar Functional Improvement After Endoprosthetic Reconstruction of Lower-Extremity Tumors.
Although the treatment of lower-extremity bone tumors is similar between adult and pediatric patients, differences in outcomes are unknown. Outcomes for lower-extremity oncologic reconstruction have been challenging to study because of the low incidence and heterogeneity in disease and patient characteristics. The PARITY (Prophylactic Antibiotic Regimens in Tumor Surgery) trial is the largest prospective data set assembled to date for patients with lower-extremity bone tumors and presents an opportunity to investigate differences in outcomes between these groups. Patient details were acquired from the prospectively collected PARITY trial database. The 1993 Musculoskeletal Tumor Society (MSTS-93) and Toronto Extremity Salvage Score (TESS) questionnaires were administered preoperatively and at 3, 6, and 12 months postoperatively. Continuous outcomes were compared between groups with use of the Student t test, and dichotomous outcomes were compared with use of the Pearson chi-square test. A total of 150 pediatric and 447 adult patients were included. Pediatric patients were more likely than adult patients to have a primary bone tumor (146 of 150 compared with 287 of 447, respectively; p < 0.001) and to have received adjuvant chemotherapy (140 of 149 compared with 195 of 441, respectively; p < 0.001). Reoperation rates were not significantly different between age groups (45 of 105 pediatric patients compared with 106 of 341 adult patients; p ≤ 0.13). Pediatric patients had higher mean MSTS-93 scores (64.7 compared with 53.8 among adult patients; p < 0.001) and TESS (73.4 compared with 60.4 among adult patients; p < 0.001) at baseline, which continued to 1 year postoperatively (mean MSTS-93 score, 82.0 compared with 76.8 among adult patients; p = 0.02; mean TESS, 87.7 compared with 78.6 among adult patients; p < 0.001). Despite the differences in outcomes between cohorts, pediatric and adult patients demonstrated similar improvement in MSTS-93 scores (mean difference, 17.4 and 20.0, respectively; p = 0.48) and TESS (mean difference, 14.1 and 14.7, respectively; p = 0.83) from baseline to 1 year postoperatively. Pediatric patients had significantly better functional outcomes than adult patients at nearly all of the included postoperative time points; however, pediatric and adult patients showed similar mean improvement in these outcomes at 1 year postoperatively. These findings may be utilized to help guide the postoperative expectations of patients undergoing oncologic reconstruction. Prognostic Level II. See Instructions for Authors for a complete description of levels of evidence.


Brain death revisited.
Until six decades ago, death was considered to be a specific point in time, referring to the moment at which life ends. With the availability of ventilators, even determining when death has occurred is becoming difficult, as cessation of life functions is often not simultaneous across organ systems. With increasing accessibility to intensive care units (ICUs) even in Tier II and Tier III cities, and the government making it mandatory to notify brain death to facilitate cadaveric organ transplants, it behooves the neurosurgeon and neurologist to totally understand the minutiae of brain death. The author reflects on his personal experience in certifying brain death, over two decades, in a quaternary care hospital. Knowing the pathophysiology of brain stem dysfunction will help the clinician better understand the rationale of the mandatory clinical tests prescribed. The necessity for an early diagnosis and the pitfalls in the clinical diagnosis of brain death, the limitations of the so-called confirmatory tests, and the concerns and ethical issues will be highlighted. Review of the world literature demonstrates that there is no international consensus even on what constitutes brain death. An individual could be considered legally dead in one country but not in another!


Human metapneumovirus infection in febrile children with lower respiratory diseases in primary care settings in Hiroshima, Japan.
Human metapneumovirus (hMPV) has been shown to be a leading cause of viral lower respiratory tract infections in children. Nevertheless, few reports regarding hMPV infections over consecutive years in children in primary care settings are available. We carried out virologic and clinical studies to determine the role of hMPV in febrile lower respiratory infections in children at a primary care clinic over 3 years and 5 months. Nasopharyngeal aspirates obtained from children with acute respiratory tract infections accompanied by high-grade fever (> or = 39 degrees C) and productive cough were studied for hMPV by reverse transcription-polymerase chain reaction and for other respiratory viruses by viral cultures and immunoassays. Of 379 patients tested, 202 were positive for at least 1 virus, including 98 with hMPV, 69 with respiratory syncytial virus, 18 with adenovirus, 12 with enterovirus, 8 with parainfluenza virus, 3 with rhinovirus, 2 with influenza virus type C, and 1 with herpes simplex virus. The male:female ratio of hMPV-infected children was 0.96:1 with an overall mean age of 3.5 years (range, 2 months to 9 years). These infections occurred predominantly from February to July, and the hospitalization rate was 4%. Of 93 patients infected with hMPV alone, 52 (56%) showed evidence of a lower respiratory tract infection.


The first demonstration of CGRP-immunoreactive fibers in canine hearts: coronary vasodilator, inotropic and chronotropic effects of CGRP in canine isolated, blood-perfused heart preparations.
CGRP-immunoreactive nerve fibers were histologically stained in the endocardium and perivascular layer of coronary vessels of canine hearts. To examine the physiological role of the CGRP in the heart function, effects of exogeneous CGRP on the hearts were studied using canine isolated, blood-perfused heart preparations. CGRP exerted dose-related potent vasodilator effects with a minimal increase in the developed tension of the papillary muscle, but slightly decreased the sinoatrial rate. The vasodilator effects were unaffected by the pretreatment of either atropine or propranolol. These specific functional effects on the coronary artery are well in accordance with the anatomical localization of CGRP. Taken together, CGRP seems to play an important role in the regulation of coronary vascular tone, while it has only a small functional role in inotropism and chronotropism in canine hearts.


Ankle dislocation without fracture.
Dislocation of the ankle without accompanying malleolar fracture has been regarded as a rare lesion, with few cases reported in the literature. To date, there has been no precedent for accurate descriptions of the mechanisms, optimum treatment, and long-term prognosis of this injury. Our goal was to evaluate these variables by a retrospective review of cases from our institution. We identified eight patients who had sustained ankle dislocation without fracture and were treated at the University of Iowa during the period 1958 to 1986. We interviewed and examined each patient and obtained ankle radiographs at an average of 11.5 years postinjury (range 2 to 24 years). After analyzing our cases and other reported in the literature, we have found that this injury is most common in young people (average age 31 years, range 10 to 73 years) and males (72%), and occurs most frequently in falls, motor vehicle accidents, and sports (86%). Medial displacement occurs most frequently (27%). Disruption of the mortise occurs variably. The most likely mechanism appears to be anterior or posterior extrusion of the talus from the mortise secondary to a force applied to the plantarflexed foot. Final displacement is then determined by the position of the foot and the direction of the force applied. Physical findings are commensurate with the deformity. Neurovascular compromise is uncommon (10%). Closed reduction is almost invariably accomplished easily unless the deformity is accompanied by posterior tibiofibular dislocation. Optimum treatment appears to be immobilization in a short leg cast for 6 weeks with no weightbearing for the first 3 weeks. Long-term follow-up revealed the following. Results were all good to excellent considering the following variables: return to work and sports activities, pain, instability, swelling, and ankle and subtalar joint motion. No patient reported instability and all returned to work and sports participation. We noted mild pain and swelling that was not severe enough to require medication in 25% of patients. Range of motion was normal in all but four patients; none of these lacked more than 10 degrees of motion in any plane. Radiographic abnormalities consisted of minor ligamentous or capsular calcification in all patients, small osteophytes in four patients, and minimal joint space narrowing in one patient. No patient had normal radiographs.


Laparoscopic antegrade cholecystectomy: a standard procedure?
Retrograde approach ("fundus first") is often used in open surgery, while in laparoscopic cholecystectomy (LC) is less frequent. LC, with antegrade access, is done by putting in traction the infundibulum and going up to the fundus before to clip the cystic. Our study analyzes a number of surgical procedures performed by experienced surgeons in laparoscopy. From 2002 to 2015, 1740 laparoscopic cholecystectomies were performed at our Institution. The operative procedure performed since 2002 consists of the incision of the visceral peritoneum from the infundibulum away from Calot's triangle along the gallbladder bed up to the fundus. Then it continues from the fundus up to the infundibulum. There were no bile duct injuries. Average operative time was 40 min. 22 conversions to an open procedure (1.3%) occurred, in cases of acute cholecystitis and cirrhotic patient. Postoperative stay was mean 2 days with no delayed sequelae on follow up. gallbladder antegrade dissection for laparoscopic cholecystectomy can reduce the time of surgery and is an easier technique to perform. Therefore, it can be proposed as the standard procedure and not only be used for difficult cholecystectomies.


Models, theory structure and mechanisms in biochemistry: The case of allosterism.
From the perspective of the new mechanistic philosophy, it has been argued that explanatory causal mechanisms in some special sciences such as biochemistry and neurobiology cannot be captured by any useful notion of theory, or at least by any standard notion. The goal of this paper is to show that a model-theoretic notion of theory, and in particular the structuralist notion of a theory-net already applied to other unified explanatory theories, adequately suits the MWC allosteric mechanism explanatory set-up. We also argue, contra some mechanistic claims questioning the use of laws in biological explanations, that the theory reconstructed in this way essentially contains non-accidental regularities that qualify as laws, and that taking into account these lawful components, it is possible to explicate the unified character of the theory. Finally, we argue that, contrary to what some mechanists also claim, functional explanations that do not fully specify the mechanistic structure are not defective or incomplete in any relevant sense, and that functional components are perfectly explanatory. The conclusion is that, as some authors have emphasized in other fields (Walmsley 2008), particular elements of traditional approaches do not contradict but rather complement the new mechanist philosophy, and taken together they may offer a more complete understanding of special sciences and the variety of explanations they provide.


Assessing objective countertransference with a computer-delivered Impact Message Inventory (IMI-C).
The circumplex version of the Impact Message Inventory (IMI-C) is one of the few existing diagnostic devices for the assessment of 'objective' (inter-subjective) countertransference. The IMI-C purports to assess eight categories of countertransference reactions along the circumference of the interpersonal circle, conceptualized as blends of the two basic dimensions of affiliation (AFF) and control (CON): dominant (D), hostile-dominant (HD), hostile (H), hostile-submissive (HS), submissive (H), friendly-submissive (FS), friendly (F) and friendly-dominant (FD). As a measure of objective (or 'inter-subjective') countertransference, a crucial test for the IMI-C is its ability to produce scores that are generalizable across different therapists involved with the same patients. In four previous psychometric studies, the 'objectivity' of countertransference reactions was examined with a Dutch paper-and-pencil IMI-C version. In the present study, the generalizability of countertransference reactions across therapists was tested with a computer-delivered IMI-C. Results obtained with the electronic IMI-C format were compared with the findings of the paper-and-pencil version. Similar to the four prior studies, D and HD were the comparatively most generalizable classes of countertransference reactions among the eight categories. Both the AFF and CON main dimensions scores were substantially generalizable across therapists. From different perspectives of analysis, more favorable results were obtained with the computer-delivered IMI-C in comparison to the paper-and-pencil-version for (a) five of the eight countertransference classes (D, HD, F, FS, FD) and also for (b) the AFF main dimension. The computer-based IMI-C appears to be a promising tool for assessing objective countertransference reactions.


Effect of trimmed asparagus by-products supplementation in broiler diets on performance, nutrients digestibility, gut ecology, and functional meat production.
Trimmed asparagus by-products (TABP) is the resultant waste from asparagus possessing. TABP has fructans, such as inulins and fructooligosaccharide, which can be utilized as an alternative prebiotic. This study was conducted to examine the effect of TABP dietary supplementation on the productive performance, nutrient digestibility, gut microbiota, volatile fatty acid (VFA) content, small-intestine histology, and meat quality of broilers. A total of 320 1-day-old broiler chicks (Ross 308®) were raised under ambient temperature and assigned through a completely randomized design to one of four dietary treatments, with four replicates per treatment. The dietary treatments comprised corn-soybean basal diet supplemented with 0 (control), 10, 30, or 50 g/kg TABP. All birds were provided drinking water and feed ad libitum to meet the standard nutritional requirements of National Research Council for broiler chickens. TABP supplementation to the broilers significantly increased the apparent ether extract, crude fiber, and gross energy digestibility (p<0.05). TABP supplementation significantly increased lactic bacteria and Enterococcus spp. numbers as well as acetic, propionic, butyric, and total VFA levels (p<0.01); on the other hand, it also significantly decreased Salmonella spp. and Escherichia coli contents in the cecum compared with the control group (p<0.01). Moreover, TABP supplementation increased villus height in the duodenum and jejunum (p<0.01), cryptal depth in the jejunum and ileum (p<0.01), and villus surface areas in the duodenum, jejunum, and ileum (p<0.01). Overall, 0-35 day TABP supplementation significantly increased the feed intake (p<0.01) and average daily gain of broilers (p<0.05), but not significantly affected the viability, productive index, and economic benefit return (p>0.05). The carcass characteristics, pH, color, and water holding capacity of the chicken meat between groups were not significantly different (p>0.05). All levels of TABP supplementation appeared to be a feasible means of producing broilers with the lower serum low-density lipoprotein cholesterol and triglyceride levels as well as atherogenic indices of serum compared with the control (p<0.05). Cholesterol contents and palmitic acid, oleic acid, saturated fatty acids, and Monounsaturated fatty acids levels decreased with an increase of TABP supplementation (p<0.05). Furthermore, TABP supplementation decreased atherogenic index (AI) and thrombogenicity index (TI) of meat (p<0.05). Supplementation of 30 g/kg TABP in broiler diet could enhance broiler performance and provide chicken meat with beneficial properties, with decreased AI and TI resulted from altered cholesterol and fatty acid profiles.


[Optimization of training: development of a new partial load mode of strength training].
Hypertrophic effect of strength training is known to originate from mechanical and metabolic stimuli. During exercise with restricted blood supply ofworking muscles, that is under conditions of intensified metabolic shifts, training effect may be achieved with much lower external loads (20% of one repetition maximum (1 RM)). The aim of the study was to compare the effects of 8 wks high-intensity (80-85% MVC) strength training and low-intensity (50% 1 RM) training without relaxation. The high-intensity strength training leads to somewhat higher increments in strength and size of trained muscles than training without relaxation. During high-intensity training an increase of area occupied by type II fibers at muscle cross section prevails while during training without relaxation - an increase of area occupied by type I fibers takes place. An exercise session without relaxation leads to a more pronounced increase in secretion of growth hormone, IGF-1 and cortisol. Expression of gene regulating myogenesis (Myostatin) is changed in different ways after high-intensity strength exercise session and after exercise session without relaxation. Low-intensity strength training (50% 1 RM) without relaxation is an effective way for inducing increases of strength and size of trained muscles. This low intensive type of training may be used in restorative medicine, sports and physical culture.


Predicting Outcomes to Optimize Disease Management in Inflammatory Bowel Diseases.
Efforts to slow or prevent the progressive course of inflammatory bowel diseases [IBD] include early and intensive monitoring and treatment of patients at higher risk for complications. It is therefore essential to identify high-risk patients - both at diagnosis and throughout disease course. As a part of an IBD Ahead initiative, we conducted a comprehensive literature review to identify predictors of long-term IBD prognosis and generate draft expert summary statements. Statements were refined at national meetings of IBD experts in 32 countries and were finalized at an international meeting in November 2014. Patients with Crohn's disease presenting at a young age or with extensive anatomical involvement, deep ulcerations, ileal/ileocolonic involvement, perianal and/or severe rectal disease or penetrating/stenosing behaviour should be regarded as high risk for complications. Patients with ulcerative colitis presenting at young age, with extensive colitis and frequent flare-ups needing steroids or hospitalization present increased risk for colectomy or future hospitalization. Smoking status, concurrent primary sclerosing cholangitis and concurrent infections may impact the course of disease. Current genetic and serological markers lack accuracy for clinical use. Simple demographic and clinical features can guide the clinician in identifying patients at higher risk for disease complications at diagnosis and throughout disease course. However, many of these risk factors have been identified retrospectively and lack validation. Appropriately powered prospective studies are required to inform algorithms that can truly predict the risk for disease progression in the individual patient.


Effects of different biotic substrata on mussel attachment.
Surface colonization by invertebrates can be stimulated or inhibited by cues produced by biofilms, conspecifics or other macroorganisms. To study the effects of living substrata on the attachment of the brown mussel, Perna perna, two different approaches were employed: (1) mussels were distributed in sets of Petri dishes consisting of one sterile set (controls), three sets in which marine biofilms were allowed to develop in aquaria for 1, 7 or 15 days and another set that had been immersed in a natural marine environment for 1-day. There was no significant effect of biofilms on attachment, suggesting that neither age nor the source of the biofilm influenced attachment. (2) Mussels were suspended over PVC panels (controls) and over panels on which Balanus trigonus (Crustacea), Schizoporella errata (Bryozoa), Symplegma rubra or Didemnum speciosum (Ascidiacea) were present. Attachment was significantly higher on the controls and on B. trigonus than on colonial taxa such as S. rubra, S. errata and D. speciosum, probably due to antifouling defenses of these species. The results show that the composition of the biological substratum is an important factor affecting mussel behavior.


Oxidative stress and antioxidant status in fetal circulation in preeclampsia.
Preeclampsia is associated with oxidative stress in maternal circulation. The purpose of this study was to explore oxidative stress and antioxidants in the fetal circulation in preeclampsia. Women with preeclampsia (n = 19) or uncomplicated pregnancies (n = 33) delivered by cesarean section were included. Blood was sampled separately from the umbilical vein and artery. 8-Iso-prostaglandin F(2alpha) (8-isoprostane), a stable product of lipid peroxidation, is a reliable marker of oxidative stress. Concentration of total 8-isoprostane in cord plasma was analyzed by gas chromatography-mass spectrometry. Antioxidant status was evaluated measuring ferric reducing ability of plasma and vitamin E. There was no difference between preeclampsia and control groups regarding median plasma concentration of 8-isoprostane in umbilical vein (955 versus 780 pg/mL, p = 0.41) or in umbilical artery (233 versus 276 pg/mL, p = 0.65). Concentration of 8-isoprostane was much higher in plasma from the umbilical vein than artery, suggesting placenta as the source of 8-isoprostane. Median ferric reducing ability of plasma concentration was higher in preeclampsia than in controls, both in the umbilical vein and artery. Median vitamin E concentration in the umbilical vein was higher in preeclampsia, but no difference was found in the umbilical artery. In conclusion, no evidence of increased oxidative stress, evaluated by 8-isoprostane concentration, was found in fetal circulation in preeclampsia.


[Evaluation of the Chêneau brace in adolescent idiopathic scoliosis].
We aimed to evaluate the Chêneau brace in the orthopedic treatment of adolescent idiopathic scoliosis to better determine the indications under which it could be prescribed. This was a retrospective study including 63 patients treated by Chêneau brace for adolescent idiopathic scoliosis between 1997 and 2006. The Cobb angles of the curves in the frontal and sagittal planes as well as rotations and rib hump were measured at the beginning of treatment, with the brace, at the end of treatment and 2 years after discontinuing the brace. A variation of+/-10 degrees in angle was selected to judge the results. At the end of the treatment, 25.4% of the curves were improved and 60.3% stabilized, with 14.3% aggravated; 5 patients (7.9%) required surgery. At 2 years, the reduction in angle was 1.8 degrees, on average. The best results were obtained for the lumbar and dorsolumbar curves. A significant improvement was noted for the dorsal curves; the major double curves are not as accessible to the treatment. For curves whose initial angle was less than 30 degrees , an initial reduction of higher than 50% with the brace presented the best improvement at the end of the follow-up. We note a significant reduction of the rib hump but not accompanied by a reduction of rotation. Finally, we note a deleterious effect in the sagittal curves, with a slight flatness of the spinal profile. The continuous wearing of the Chêneau brace can stabilize the evolution of adolescent idiopathic scoliosis, apart from the forms with major double curves, which are not as accessible to the treatment. However, the improvement comes at the price of a slight flatness of the curves of the spine in the sagittal plane. The loss of the correction after discontinuing the brace is minor. We emphasize, therefore, the necessity to monitor the sagittal aspect of the spine as well as the possibility of early treatment, strict surveillance and prolonged follow-up.


Palliative radiation therapy for Kaposi's sarcoma in patients with AIDS.
Kaposi's sarcoma is a common manifestation of AIDS, particularly in young male homosexual patients. Between 1987 and 1988, eight patients with Epidemic Kaposi's sarcoma received palliative radiation therapy. The authors' experience indicates that radiation therapy can provide good to excellent palliation for skin and oropharyngeal lesions.


A Sweet Paradox: Severe Insulin Resistance and Hyperglycemia in Asymptomatic COVID-19 Infection.
There is a well-established association between hyperglycemia and severe coronavirus disease 2019 (COVID-19) infection, regardless of the diagnosis of diabetes prior to the infection. However, it is unusual for patients with a mild infection to present with severe hyperglycemia and insulin resistance requiring intravenous insulin therapy. Uncontrolled hyperglycemia is associated with worse outcomes in COVID-19, making it crucial to achieve optimal glycemic control, which occasionally requires IV insulin therapy. We report a patient with type 1 diabetes mellitus (T1DM), on hemodialysis, who presented with diabetic ketoacidosis (DKA) due to non-adherence to insulin. He was found to be incidentally positive for COVID-19 on admission. Although he was asymptomatic and did not require steroids for the treatment of COVID-19, he was noted to have persistent severe hyperglycemia requiring unusually high levels of intravenous insulin. This proposes that even a mild infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection can trigger a systemic response that can lead to downstream manifestations including insulin resistance and severe hyperglycemia. Interestingly, our patient had three admissions within the past six months as well as another admission two weeks after the current presentation with DKA secondary to insulin non-compliance, all of which required IV insulin for <24 hours following which he was transitioned to a basal-bolus insulin regimen with well-controlled glucose levels.


Selection and validation of potato candidate genes for maturity corrected resistance to Phytophthora infestans based on differential expression combined with SNP association and linkage mapping.
Late blight of potato (Solanum tuberosum L.) caused by the oomycete Phytophthora infestans (Mont.) de Bary, is one of the most important bottlenecks of potato production worldwide. Cultivars with high levels of durable, race unspecific, quantitative resistance are part of a solution to this problem. However, breeding for quantitative resistance is hampered by the correlation between resistance and late plant maturity, which is an undesirable agricultural attribute. The objectives of our research are (i) the identification of genes that condition quantitative resistance to P. infestans not compromised by late plant maturity and (ii) the discovery of diagnostic single nucleotide polymorphism (SNP) markers to be used as molecular tools to increase efficiency and precision of resistance breeding. Twenty two novel candidate genes were selected based on comparative transcript profiling by SuperSAGE (serial analysis of gene expression) in groups of plants with contrasting levels of maturity corrected resistance (MCR). Reproducibility of differential expression was tested by quantitative real time PCR and allele specific pyrosequencing in four new sets of genotype pools with contrasting late blight resistance levels, at three infection time points and in three independent infection experiments. Reproducibility of expression patterns ranged from 28 to 97%. Association mapping in a panel of 184 tetraploid cultivars identified SNPs in five candidate genes that were associated with MCR. These SNPs can be used in marker-assisted resistance breeding. Linkage mapping in two half-sib families (n = 111) identified SNPs in three candidate genes that were linked with MCR. The differentially expressed genes that showed association and/or linkage with MCR putatively function in phytosterol synthesis, fatty acid synthesis, asparagine synthesis, chlorophyll synthesis, cell wall modification, and in the response to pathogen elicitors.


Differences on the effect of heat waves on mortality by sociodemographic and urban landscape characteristics.
Mortality increases during heat waves have been reported worldwide. The magnitude of these increases can vary within regions according to sociodemographic and urban landscape characteristics. The objectives of this study were to explore this variation and its determinants, and to identify the most heat-vulnerable areas by mapping heat vulnerability. We conducted a time-stratified case-crossover analysis using daily mortality in the Barcelona metropolitan area during the warm seasons of 1999-2006. Temperature data on the date of death were assigned to each individual, which were assigned to their census tract of residence. Eight census tract-level variables on socioeconomic or built environment characteristics were obtained from the census. Residence surrounding greenness was obtained from satellite data. The relative risk (RR) of mortality after three consecutive hot days (defined as those exceeding the 95th percentile of maximum temperature) was calculated via conditional logistic regression. Effect modification was examined by including interaction terms. Analyses were based on 52 806 deaths. The effect of three consecutive hot days was a 30% increase in all-cause mortality (RR=1.30, 95% CI 1.24 to 1.38). Heterogeneity of this effect was observed across census tracts. The effect of heat on mortality was higher in the census tracts with a large percentage of old buildings (RR=1.21, 95% CI 1.00 to 1.46), manual workers (RR=1.25, 95% CI 0.96 to 1.64) and residents perceiving little surrounding greenness (RR=1.29, 95% CI 1.01 to 1.65). After three consecutive hot days, mortality doubled in the most heat-vulnerable census tracts. Sociodemographic and urban landscape characteristics are associated to mortality risk during heat waves and are useful to build heat vulnerability maps.


Risk factors for mortality in patients with Staphylococcus aureus bloodstream infection.
In this two year retrospective analysis, we evaluated the epidemiology and risk factors for mortality of Staphylococcus aureus bloodstream infection (SaBSI). Methicillin-susceptible S. aureus (MSSA) was isolated in 84 (44.2%) and methicillin-resistant S. aureus (MRSA) in 106 episodes (55.8%). The mortality rate after 21 days was 16.4%. At univariate analysis older age, no removal of central venous catheter (CVC), prosthetic heart valves, severe sepsis, septic shock and high APACHE II score were significantly associated with mortality, whereas treatment duration > 48 hours, appropriate targeted therapy and prolonged treatment duration were significantly associated with survival. At multivariate analysis, prosthetic valves, septic shock and fever 48 hours after the diagnosis were significantly related to mortality. In this study, the mortality was associated with clinical rather than microbiological factors.


Comparison of Standard Culture-Based Method to Culture-Independent Method for Evaluation of Hygiene Effects on the Hand Microbiome.
Hands play a critical role in the transmission of microbiota on one's own body, between individuals, and on environmental surfaces. Effectively measuring the composition of the hand microbiome is important to hand hygiene science, which has implications for human health. Hand hygiene products are evaluated using standard culture-based methods, but standard test methods for culture-independent microbiome characterization are lacking. We sampled the hands of 50 participants using swab-based and glove-based methods prior to and following four hand hygiene treatments (using a nonantimicrobial hand wash, alcohol-based hand sanitizer [ABHS], a 70% ethanol solution, or tap water). We compared results among culture plate counts, 16S rRNA gene sequencing of DNA extracted directly from hands, and sequencing of DNA extracted from culture plates. Glove-based sampling yielded higher numbers of unique operational taxonomic units (OTUs) but had less diversity in bacterial community composition than swab-based sampling. We detected treatment-induced changes in diversity only by using swab-based samples (P < 0.001); we were unable to detect changes with glove-based samples. Bacterial cell counts significantly decreased with use of the ABHS (P < 0.05) and ethanol control (P < 0.05). Skin hydration at baseline correlated with bacterial abundances, bacterial community composition, pH, and redness across subjects. The importance of the method choice was substantial. These findings are important to ensure improvement of hand hygiene industry methods and for future hand microbiome studies. On the basis of our results and previously published studies, we propose recommendations for best practices in hand microbiome research.IMPORTANCE The hand microbiome is a critical area of research for diverse fields, such as public health and forensics. The suitability of culture-independent methods for assessing effects of hygiene products on microbiota has not been demonstrated. This is the first controlled laboratory clinical hand study to have compared traditional hand hygiene test methods with newer culture-independent characterization methods typically used by skin microbiologists. This study resulted in recommendations for hand hygiene product testing, development of methods, and future hand skin microbiome research. It also demonstrated the importance of inclusion of skin physiological metadata in skin microbiome research, which is atypical for skin microbiome studies.


Evaluation of the Safety and Efficacy of Conventional Transarterial Chemoembolization (cTACE) and Drug-Eluting Bead (DEB)-TACE in the Management of Unresectable Hepatocellular Carcinoma: A Systematic Review.
Transarterial chemoembolization (TACE) is considered the preferred loco-regional treatment option for hepatocellular carcinoma (HCC) not amenable to resection due to its distinctive blend of precise drug administration, localized tumor management, and reduced systemic adverse effects, setting it apart from the plethora of alternative treatments available. There is an ongoing debate regarding the optimal choice for managing HCC using TACE, particularly between its two major types: conventional TACE (cTACE) and drug-eluting bead TACE (DEB-TACE). The medical community remains divided on which approach offers superior safety and efficacy, with conflicting evidence and varied outcomes adding to the complexity of this nuanced decision. Given the lack of consensus surrounding the preferred TACE technique in treatment-naive patients for HCC, we conducted a rigorous systematic review to assess and contrast the relative safety and efficacy of cTACE versus DEB-TACE in patients diagnosed with HCC who did not receive any prior treatment for HCC. Our study aimed to provide much-needed clarity on this controversial topic, shedding light on the two approaches' comparative safety and efficacy to inform clinical decision-making. After a comprehensive search of databases and search engines and through a methodical screening process, including standardized quality assessments and relevant filter application based on our eligibility criteria, we identified 10 articles pertinent to our research query comprising two randomized controlled trials, one meta-analysis, and seven observational studies. The collective sample size of the studies was 5,288 patients with HCC, of which 2,959 were in the cTACE arm and 2,324 were in the DEB-TACE arm.


Workforce planning. Of one mind.
The NHS is heavily reliant on locum doctors, many of whom trained overseas. It is important they are appropriately trained so they can be integrated into the workforce. A pilot nine-session course for consultant locum psychiatrists was well received by participants. Medical directors have expressed willingness to pay for locums to attend such courses.


Advances in genetic manipulation of Chlamydia trachomatis.
Chlamydia trachomatis, one species of Chlamydia spp., has the greatest impact on human health and is the main cause of bacterial sexually transmitted diseases and preventable blindness among all Chamydia spp. species. The obligate intracellular parasitism and unique biphasic developmental cycle of C. trachomatis are the main barriers for the development of tools of genetic manipulation. The past decade has witnessed significant gains in genetic manipulation of C. trachomatis, including chemical mutagenesis, group II intron-based targeted gene knockout, fluorescence-reported allelic exchange mutagenesis (FRAEM), CRISPR interference (CRISPRi) and the recently developed transposon mutagenesis. In this review, we discuss the current status of genetic manipulations of C. trachomatis and highlights new challenges in the nascent field of Chlamydia genetics.


Up-regulation of IL-18 and IL-12 in the ileum of neonatal rats with necrotizing enterocolitis.
Necrotizing enterocolitis (NEC) is a common and devastating gastrointestinal disease of premature infants. Because the proinflammatory cytokines IL-18, IL-12, and interferon (IFN)-gamma have been implicated in other diseases of the small intestine, we hypothesized that these cytokines would play an important role in NEC pathogenesis. NEC was induced in newborn rats via enteral feeding with rat milk substitute and asphyxia and cold stress (RMS). Dam-fed, asphyxia- and cold-stressed littermates were used as controls (DF). After 96 h, the distal ileum was removed from all animals and processed to determine expression and localization of IL-18, IL-12, and IFN-gamma using real-time reverse transcriptase PCR and immunohistology. IL-18 and IL-12 mRNA from the RMS group were increased (p < or = 0.05) compared with DF controls, and there was a correlation between increasing IL-18 and IL-12 mRNA levels and progression of tissue damage (r = 0.629 and 0.588, respectively; p < or = 0.05). Immunohistology revealed IL-18 in the cytoplasm of villi and crypt enterocytes and IL-12-positive monocytes/macrophages were increased with disease progression (r = 0.503, p < or = 0.05). No differences in the number of IFN-gamma-positive cells were observed between groups. These data demonstrate up-regulation of IL-18 and IL-12 in experimental NEC and a correlation between production of these proinflammatory cytokines and progression of tissue damage.


Confirmation and quantification of clenbuterol in horse urine using liquid chromatography tandem mass spectrometry triple quadrupole.
Clenbuterol (CLE) is used in horses as a bronchodilator and for its anabolic steroid-like effects. CLE is a Class 3 drug according to current Association of Racing Commissioners International (ARCI) Uniform Classification Guidelines. The Racing Medication and Testing Consortium recommended a urine CLE threshold of 140 pg/mL after careful scientific review of the results of studies describing the disposition of CLE in the horse and this threshold was adopted by the ARCI. Enzyme-linked immunosorbent assay was previously used to screen samples for CLE in Illinois, but could not detect such low concentrations in urine. Thus, a liquid-liquid extraction of CLE from urine followed by quantification by liquid chromatography-tandem mass spectrometry was developed and validated. Method validation included testing stability, ion suppression and enhancement, precision, accuracy and uncertainty. Intra-, interday and total precision and accuracy were calculated for each control and found to be within the ±15% acceptance range. The Guide to the Expression of Uncertainty in Measurement approach was used to calculate uncertainty, which was 11% at the 95% confidence level. In the past 5 years, only 15 samples were reported as positive for CLE in Illinois. This new method was used in a pilot program to screen and confirm samples received from thoroughbred and harness horses.


Enzymatic hydrolysis and fermentation of pretreated cashew apple bagasse with alkali and diluted sulfuric Acid for bioethanol production.
The aim of this work was to optimize the enzymatic hydrolysis of the cellulose fraction of cashew apple bagasse (CAB) after diluted acid (CAB-H) and alkali pretreatment (CAB-OH), and to evaluate its fermentation to ethanol using Saccharomyces cerevisiae. Glucose conversion of 82 +/- 2 mg/g CAB-H and 730 +/- 20 mg/g CAB-OH was obtained when 2% (w/v) of solid and 30 FPU/g bagasse was used during hydrolysis at 45 degrees C, 2-fold higher than when using 15 FPU/g bagasse, 44 +/- 2 mg/g CAB-H, and 450 +/- 50 mg/g CAB-OH, respectively. Ethanol concentration and productivity, achieved after 6 h of fermentation, were 20.0 +/- 0.2 g L(-1) and 3.33 g L(-1) h(-1), respectively, when using CAB-OH hydrolyzate (initial glucose concentration of 52.4 g L(-1)). For CAB-H hydrolyzate (initial glucose concentration of 17.4 g L(-1)), ethanol concentration and productivity were 8.2 +/- 0.1 g L(-1) and 2.7 g L(-1) h(-1) in 3 h, respectively. Hydrolyzates fermentation resulted in an ethanol yield of 0.38 and 0.47 g/g glucose with pretreated CAB-OH and CAB-H, respectively. Ethanol concentration and productivity, obtained using CAB-OH hydrolyzate, were close to the values obtained in the conventional ethanol fermentation of cashew apple juice or sugar cane juice.


The Influence of the Washing Process on the Impedance of Textronic Radio Frequency Identification Transponder Antennas.
Antennas dedicated to RFID systems created on textile substrates should maintain strictly defined parameters. During washing, the materials from which such antennas are made are exposed to mechanical and chemical exposure-degradation of the parameters characterizing those materials may occur, which in turn may lead to a change in the parameters of the antenna. For research purposes, four groups of model dipole antennas (sewn with two types of conductive threads on two fabrics) were created and then they were subjected to several washing processes. After each stage of the experiment, the impedance parameters of the demonstration antennas were measured using indirect measurements. Based on the obtained results, it was found that these parameters change their values during washing, and that this is influenced by a number of factors, e.g., shrinkage of the substrate fabric.


Serous microcystic adenoma (glycogen-rich cystadenoma) of the pancreas: study of 11 cases showing clinicopathological and immunohistochemical correlations.
Serous microcystic adenoma of the pancreas, also known as microcystic adenoma, glycogen-rich cystadenoma or serous cystadenoma, is an uncommon benign tumor. We have studied 11 cases involving eight women and three men. The average age at diagnosis was 61.7 years. Four tumors were discovered incidentally. Tumors varied from 1.2 to 20 cm in maximum diameter and all were multicystic. Within the pancreas, three were located in the pancreas head, one involved the head and body, one was located in the body, five were in the tail, and one occupied the whole pancreas. Central stellate scar was seen in five (45%) cases. Histologically, all tumors were composed of microglandular cysts lined by clear epithelial cells rich in glycogen, which were separated by fibrocollagenous stroma. The expression of keratin in clear epithelial cells resembled that in ductal and/or centroacinar cells, but not acinar cells. alpha-Smooth muscle actin (SMA)-positive myoepithelial cells and stromal amyloid deposits were not detected. Ultrastructurally, fibrocollagenous stroma was composed of alpha-SMA-positive myofibroblasts and endothelial cells embedded in thick collagen bundles. Regardless of female propensity, estrogen and progesterone receptors were not detected. Therefore, female predominance in this tumor remains to be elucidated.


Sequence analysis of the ribosomal DNA intergenic spacer 1 regions of Trichosporon species.
We determined the sequence of the intergenic spacer (IGS) 1 region, which is located between the 26S and 5S rRNA genes, in 25 species of the genus TRICHOSPORON: IGS 1 sequences varied in length from 195 to 719 bp. Comparative sequence analysis suggested that the divergence of IGS 1 sequences has been greater than that of the internal transcribed spacer regions. We also identified five genotypes of T. asahii, which is a major causative agent of deep-seated trichosporonosis, based on the IGS 1 sequences of 43 strains. Most of the isolates that originated in Japan were of genotype 1, whereas the American isolates were of genotype 3 or 5. Our results suggest that analysis of IGS regions provides a powerful method to distinguish between phylogenetically closely related species and that a geographic substructure may exist among T. asahii clinical isolates.


[Injuries among Sapienza University students].
Background. The phenomenon of accidents during the university course is a public health problem. Students spend a lot of time at university and in structure for training. This this leads them to be exposed to biological, chemical and ergonomic risk of accidents or occupational diseases. Aim. The aim of this study is to describe accidents that occurred to students of Sapienza University in Rome during the period 2010-2015. Materials and Methods. Retrospective study conducted on a cohort of students from Sapienza University in the years 2010-2015. As a source of data was used register of accidents reported by the students. Results. During the period considered, injuries were recorded in 791 students, mainly female (71.8%). Nursing students were the most affected (92.8%), followed by medical students (5.7%) and finally students of other faculties (1.5%). Point wounds were the most frequent (n=462) representing 58.4% of the total and occur mainly at the wrist and hand (n=380); the second most frequent type of injury is contact with biological liquid (n=159) followed by contusions (n=72), the most affected site in this case are the head and the face (n=81; n=23). Contusions often also involve multiple regions (n=17). In lesser numbers there are also the distortions (n=55), and fractures (n=26) the most frequent first ankle and foot (25.5%), the second most frequent wrist and hand (34.6%) The incidence of accidents ranged from 0.079% in 2011 to 0.161% in 2012. Conclusion. There is a need to design measures for more intensive specific training on biomedical students, especially nursing students, as the most exposed category, in order to reduce the accident phenomenon. Particular attention should be paid to the safe handling of sharp objects. It would be beneficial to include in the course of teaching lessons aimed at security and greater awareness of the risk.


The DNA-Dialogue: A Broad Societal Dialogue About Human Germline Genome Editing in the Netherlands.
For years, calls for public involvement in the debate concerning the acceptability of human germline genome editing (HGGE) have been made. A multidisciplinary consortium of 11 organizations in the Netherlands organized a broad societal dialogue to inquire about the views of Dutch society toward HGGE. The project aimed to reach a wide and diverse audience and to stimulate a collective process of deliberative opinion forming and reflection. To that end, several instruments and formats were developed and employed. We present the results of 27 moderated dialogues organized between October 2019 and October 2020. Overall, participants of the dialogues were capable of assessing and discussing the subject of HGGE in a nuanced way. Analysis of these dialogues shows that in general, participants had no fundamental and absolute objections toward HGGE technology. However, they only deemed HGGE to be acceptable when it is used to prevent serious heritable diseases and under strict conditions, without affecting important (societal) values. There was a small group of participants who found HGGE fundamentally unacceptable because it would cross natural, socio-ethical, or religious boundaries.


Leukocyte-induced vascular protein leakage in cat mesentery.
The overall objective of this study was to determine whether leukocyte adherence and/or emigration is a prerequisite for the increased vascular protein leakage associated with acute inflammation. An in vivo preparation was used to monitor intestinal vascular protein leakage as well as polymorphonuclear leukocyte (PMN) adhesion and emigration in feline mesenteric microvessels exposed to platelet-activating factor (PAF) and leukotriene B4 (LTB4). Local intra-arterial infusion of PAF (4 ng/min) produced a fourfold increase in vascular protein leakage. A 50-fold higher concentration of LTB4 had no effect on vascular protein efflux. LTB4, however, did potentiate the PAF-induced vascular protein leakage. Both inflammatory mediators caused leukocytes to adhere to endothelial cells in postcapillary venules; however, leukocyte emigration was observed only in the presence of PAF. PAF-induced leukocyte adhesion and emigration and the increased vascular protein leakage were inhibited by a monoclonal antibody (MoAb IB4) directed against the common beta-subunit of the adhesive glycoprotein complex CD11/CD18. MoAb IB4 also prevented LTB4-induced leukocyte adhesion. Both PAF and LTB4 caused degranulation of cat PMNs in vitro, yet superoxide production was stimulated by PAF only. The data derived from these in vivo and in vitro studies indicate that leukocyte adhesion per se does not necessarily lead to increased vascular protein leakage and leukocyte emigration. Adhesion-dependent PMN functions such as emigration and superoxide production may play an important role in producing the alterations in vascular integrity observed in inflamed microvessels.


Differential attentional bias in generalized anxiety disorder and panic disorder.
Cognitive theorists relate anxiety disorders to the way in which emotional information is processed. The existing research suggests that patients with anxiety disorders tend to allocate their attention toward threat-related information selectively, and this may differ among different types of anxious subjects. The aim of this study was to explore attentional bias in patients with generalized anxiety disorder (GAD) and panic disorder (PD) using the emotional Stroop task and compare the differences between them. Forty-two patients with GAD, 34 patients with PD, and 46 healthy controls performed the emotional Stroop task with four word types, ie, GAD-related words, PD-related words, neutral words, and positive words. Patients with GAD and those with PD were slower than healthy controls to respond to all stimuli. Patients with GAD had longer response latencies in color-naming both PD-relevant words and GAD relevant words. Patients with PD had longer response latencies only in color-naming PD-related words, similar to healthy controls. Patients with GAD and those with PD had a different pattern of attentional bias, and there was insufficient evidence to support the existence of specific attentional bias in patients with PD.


Assessment of the microbial interplay during anaerobic co-digestion of wastewater sludge using common components analysis.
Anaerobic digestion (AD) is used to minimize solid waste while producing biogas by the action of microorganisms. To give an insight into the underlying microbial dynamics in anaerobic digesters, we investigated two different AD systems (wastewater sludge mixed with either fish or grass waste). The microbial activity was characterized by 16S RNA sequencing. 16S data is sparse and dispersed, and existent data analysis methods do not take into account this complexity nor the potential microbial interactions. In this line, we proposed a data pre-processing pipeline addressing these issues while not restricting only to the most abundant microorganisms. The data were analyzed by Common Components Analysis (CCA) to decipher the effect of substrate composition on the microorganisms. CCA results hinted the relationships between the microorganisms responding similarly to the AD physicochemical parameters. Thus, in overall, CCA allowed a better understanding of the inter-species interactions within microbial communities.


Are stereotaxic breast biopsies adequate?
Stereotaxic core breast biopsy (SCBB) has been proposed as a cost-effective and reliable method of evaluating mammographic lesions. This study evaluates an initial experience with SCBB and assesses the adequacy of the biopsy specimens obtained. Two hundred forty-one SCBB were performed on 221 patients during 13 months by four radiologists. Mammograms were assigned a suspicion index on a scale of 1 to 5. One pathologist performed a blinded retrospective review of all SCBB specimens and assigned an adequacy score based on the quality and amount of the tissue present. The majority of SCBB were ordered by general surgeons (67%). A suspicion index score of 3 was assigned to 74% of lesion specimens. Twelve percent of specimens were malignant. Overall SCBB adequacy (score > or = 2) was 77%. Adequacy was present in 74% of benign biopsy specimens as compared with 100% of malignant specimens (p < 0.005). Only 62% of specimens reported as benign without specific features were adequate. There were no differences in adequacy between individual radiologists or during the study period. SCBB is largely used by surgeons to assess indeterminate mammographic lesions. One of four benign specimens was inadequate. Benign SCBB specimens must be interpreted with caution.


Evaluation of the diurnal variation in the pharmacokinetics of zileuton in healthy volunteers.
The diurnal variation in the pharmacokinetic parameters of zileuton were evaluated in 12 healthy male volunteers in a three-period study. Periods I and II constituted a balanced, randomized, crossover study in which a participant received a single dose of 600-mg zileuton either at 7 AM or 11 PM. In period III all participants received 600-mg doses four times daily for 5 days. The differences between the pharmacokinetics of single doses of zileuton administered at 7 AM and 11 PM were not statistically significant. Plasma concentration-time profiles of zileuton during the four daily dose intervals at steady state were also similar. Values for the pharmacokinetic parameters of zileuton after multiple doses were similar to those after single doses, with a minimal accumulation of the drug after multiple doses. Overall, there was little or no diurnal variation in the pharmacokinetic parameters of zileuton after single and multiple doses.


[Non-invasive imaging tests for detecting coronary artery disease in symptomatic stable patients. Is diagnostic performance sufficient for guiding strategy?].
Several non-invasive imaging techniques are currently available for coronary artery disease detection in stable patients with chest pain: exercise electrocardiogram, myocardial scintigraphy, stress echocardiography, stress MRI, positron emission tomography and computed tomography coronary angiography. According to recent guidelines from the European Society of Cardiology, the diagnosis process shall be guided by the coronary risk of the patient. The first recommended step is to clinically assess the probability of coronary artery disease. Thereafter, the choice of technique will be driven by usual parameters such as availability, local expertise and the contraindications of each test. Although detection of coronary artery disease by non-invasive tests follows different pathophysiological pathways, diagnostic value appears comparable. Therefore, choice of a diagnostic test must also take into consideration other factors such as the risks and hazards of imaging techniques as well as cost-efficiency parameters.


The Gap Between Adaptive Behavior and Intelligence in Autism Persists into Young Adulthood and is Linked to Psychiatric Co-morbidities.
For individuals with autism spectrum disorder (ASD), long-term outcomes have been troubling, and intact IQ has not been shown to be protective. Nevertheless, relatively little research into adaptive functioning among adults with ASD has been completed to date. Therefore, both adaptive functioning and comorbid psychopathology were assessed among 52 adults with ASD without intellectual disability (ID). Adaptive functioning was found to substantially lag behind IQ, and socialization was a particular weakness. Comorbid psychopathology was significantly correlated with the size of IQ-adaptive functioning discrepancy. These findings emphasize key intervention targets of both adaptive skill and psychopathology for transition-age youth and young adults with ASD, as well as the need for ongoing monitoring of anxiety and depression symptoms during this developmental window.


A single-dose comparison of inhaled albuterol and two formulations of salmeterol on airway reactivity in asthmatic subjects.
Salmeterol is a new beta 2-selective agonist with a long duration of action. We compared the effect of 50 micrograms salmeterol administered by metered dose inhaler (MDI) and dry powder inhaler (DPI), with 200 micrograms albuterol (by MDI) on FEV1 and histamine-induced bronchoconstriction in 12 asthmatic subjects in a double-blind, placebo-controlled crossover study. Subjects were studied for 12 h on four occasions with a histamine challenge test at 1, 4, 8, and 12 h, the response being measured as the dose of histamine causing a 20% fall in FEV1 (PD20). There was no difference in the mean +/- standard error of the mean maximum increase in FEV1 between salmeterol MDI, salmeterol DPI, or albuterol (0.57 +/- 0.078, 0.54 +/- 0.089, and 0.49 +/- 0.081 L, respectively); the increase following placebo was 0.25 +/- 0.082 L. The area under the 12-h time-response curve for FEV1 was greater for both formulations of salmeterol (MDI and DPI) than for albuterol. The three active drugs protected against histamine-induced bronchoconstriction. The increase in PD20 at 1 and 12 h and the area under the 12-h time-response curve for PD20 were greater following both formulations of salmeterol than after albuterol. The greater change in geometric mean PD20 values at 1 h after both formulations of salmeterol (2.21 and 1.88 mumol) relative to albuterol (0.98 mumol) suggests that despite similar effects on FEV1 the doses used do not have an equivalent effect on the airways.(ABSTRACT TRUNCATED AT 250 WORDS)


Itch E3 ubiquitin ligase positively regulates TGF-β signaling to EMT via Smad7 ubiquitination.
TGF-β regulates pleiotropic cellular responses including cell growth, differentiation, migration, apoptosis, extracellular matrix production, and many other biological processes. Although non-Smad signaling pathways are being increasingly reported to play many roles in TGF-β-mediated biological processes, Smads, especially receptor-regulated Smads (R-Smads), still play a central mediatory role in TGF-β signaling for epithelial-mesenchymal transition. Thus, the biological activities of R-Smads are tightly regulated at multiple points. Inhibitory Smad (I-Smad also called Smad7) acts as a critical endogenous negative feedback regulator of Smad-signaling pathways by inhibiting R-Smad phosphorylation and by inducing activated type I TGF-β receptor degradation. Roles played by Smad7 in health and disease are being increasingly reported, but the molecular mechanisms that regulate Smad7 are not well understood. In this study, we show that E3 ubiquitin ligase Itch acts as a positive regulator of TGF-β signaling and of subsequent EMT-related gene expression. Interestingly, the Itch-mediated positive regulation of TGF-β signaling was found to be dependent on Smad7 ubiquitination and its subsequent degradation. Further study revealed Itch acts as an E3 ubiquitin ligase for Smad7 polyubiquitination, and thus, that Itch is an important regulator of Smad7 activity and a positive regulator of TGF-β signaling and of TGF-β-mediated biological processes. Accordingly, the study uncovers a novel regulatory mechanism whereby Smad7 is controlled by Itch.


Gcn5-mediated acetylation at MBF-regulated promoters induces the G1/S transcriptional wave.
In fission yeast, MBF-dependent transcription is inactivated at the end of S phase through a negative feedback loop that involves the co-repressors, Yox1 and Nrm1. Although this repression system is well known, the molecular mechanisms involved in MBF activation remain largely unknown. Compacted chromatin constitutes a barrier to activators accessing promoters. Here, we show that chromatin regulation plays a key role in activating MBF-dependent transcription. Gcn5, a part of the SAGA complex, binds to MBF-regulated promoters through the MBF co-activator Rep2 in a cell cycle-dependent manner and in a reverse correlation to the binding of the MBF co-repressors, Nrm1 or Yox1. We propose that the co-repressors function as physical barriers to SAGA recruitment onto MBF promoters. We also show that Gcn5 acetylates specific lysine residues on histone H3 in a cell cycle-regulated manner. Furthermore, either in a gcn5 mutant or in a strain in which histone H3 is kept in an unacetylated form, MBF-dependent transcription is downregulated. In summary, Gcn5 is required for the full activation and correct timing of MBF-regulated gene transcription.


Effects of cigarette smoking with different tar content on erythrocyte oxidant/antioxidant status.
In this study, the effects of cigarettes with differing tar content on erythrocyte oxidant/antioxidant status was investigated. Malondialdehyde (MDA) as an indicator of oxidant status and superoxide radical scavenger activity (SSA) as an indicator of antioxidant status were measured in erythrocytes from 20 smokers and 10 non-smoker controls. Ten of the 20 smoking subjects smoked five cigarettes with full flavour low tar (FFLT with 12 mg tar) and the others smoked five cigarettes with full flavour high tar (FF with 23 mg tar) over 1 hour. Initial blood samples from both groups at fasting, followed by further samples from smokers at 1.5 hours and 3 hours after smoking. Initial erythrocyte MDA level and SSA activity were found to be higher in the smoking groups compared to non-smokers. Furthermore, both parameters were significantly higher at the 1.5-hour and 3-hour erythrocyte samples when compared to initial values in the FFLT group. However, there were no statistically significant differences between SSA values established at different times in FF group. Results suggest that smoking causes oxidant load in the erythrocytes. Although a compensatory mechanism (i.e. increased SSA activities) develops in the FFLT group after smoking, this cannot prevent peroxidation reactions (i.e. increased MDA levels) in the erythrocytes. As to the types of cigarettes, both seem to have oxidant potential, but oxidation degree in the FFLT group is higher than that of FF group. These results suggest that antioxidant supplementation to smokers might be beneficial to decrease cellular oxidation damages.


A new material for tissue engineered vagina reconstruction: Acellular porcine vagina matrix.
Acellular matrix materials have been widely used to repair various tissues and organs. According to the plastic principle, when a part of the body is lost, it should be replaced with a similar material. Therefore, the use of a homologous organ-specific acellular vaginal tissue in vagina reconstruction repair surgery may show good results. However, the acellular vagina matrix (AVM) form large vertebrates is difficult to isolate. In this study, we described a multistep method to prepare porcine AVM and evaluated the efficacy of acellularization. We also investigated the biomechanical properties, biological activity elements, and biocompatibility of the porcine AVM. We then used this material to reconstruct a rat vagina and performed further morphologic and functional analyses. Small intestinal submucosa (SIS), which is a commonly used acellular matrix material, was used in a control group. Histological examination, DNA content analysis, and agarose gel electrophoresis revealed that the decellularization procedure was effective. The AVM had acceptable biomechanical properties and sufficient growth factor production (VEGF, FGF, TGF-β1, and PDGF-BB) compared with that of the SIS. Subcutaneous transplantation in rats showed that the AVM had good biocompatibility. The tissue-engineered vagina using the AVM more resembled normal-appearing tissue than did that using SIS following morphologic and functional analyses. The AVM has great potential for application in vaginal reconstructive surgery. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1949-1959, 2017.


Biostimulants in Corn Cultivation as a Means to Alleviate the Impacts of Irregular Water Regimes Induced by Climate Change.
Climate change alters regular weather seasonality. Corn is one of the main crops affected by irregular water regimes. Due to complications in decision-making processes related to climate change, it is estimated that planting corn outside the optimal window results in around USD 340 million in losses per year in the United States' Corn Belt. In turn, exogenous plant growth regulators have been gaining prominence due to their potential to positively influence the morphology and physiology of plants under stress. This study was based on the hypothesis that the use of plant growth regulators can assist in mitigating the adverse effects of climate change on corn plants sown both inside and outside the recommended planting period. In this context, the effects of biostimulant application on gas exchange in corn plants sown within and outside the recommended period were evaluated. The experiment was carried out in randomized blocks in a 4 × 5 × 2 factorial scheme with four repetitions. These were four sowing times, the application of the biostimulants via seeds in five doses, and foliar applications (presence and absence). The biostimulant doses were 0.00, 6.25, 12.50, 18.75, and 25 mL kg-1. The foliar application used a dose of 500 mL ha-1. Only in the period (2017/2) higher doses of biostimulants indicated a decrease in the water use efficiency of plants, suggesting the need to evaluate this variable carefully. In this regard, future studies may investigate the ideal doses and application timings of biostimulants for different edaphoclimatic conditions. In general, the combined use of biostimulants on seeds and as a foliar treatment boosted physiological activity and stimulated photosynthetic processes in corn plants. Based on these data, plant regulators can be a useful tool to mitigate the adverse effects of climate change on corn plants sown inside and outside the planting period.


Emergence of multiple tori structures in a single polyelectrolyte chain.
We investigated the collapsed structure of a weakly charged wormlike chain under a moderate concentration of 1:1 electrolyte solution. By assuming a torus as a grand state, we found that the size of a torus is determined by the balance between surface energy and electrostatic energy, which leads to a finite torus thickness almost independent of the chain contour length. Owing to this unique characteristic, a long charged wormlike chain forms multiple tori structure as a collapsed product, which is never seen with a neutral wormlike chain. These features were confirmed by a Monte Carlo simulation.


BioVLAB-mCpG-SNP-EXPRESS: A system for multi-level and multi-perspective analysis and exploration of DNA methylation, sequence variation (SNPs), and gene expression from multi-omics data.
Measuring gene expression, DNA sequence variation, and DNA methylation status is routinely done using high throughput sequencing technologies. To analyze such multi-omics data and explore relationships, reliable bioinformatics systems are much needed. Existing systems are either for exploring curated data or for processing omics data in the form of a library such as R. Thus scientists have much difficulty in investigating relationships among gene expression, DNA sequence variation, and DNA methylation using multi-omics data. In this study, we report a system called BioVLAB-mCpG-SNP-EXPRESS for the integrated analysis of DNA methylation, sequence variation (SNPs), and gene expression for distinguishing cellular phenotypes at the pairwise and multiple phenotype levels. The system can be deployed on either the Amazon cloud or a publicly available high-performance computing node, and the data analysis and exploration of the analysis result can be conveniently done using a web-based interface. In order to alleviate analysis complexity, all the process are fully automated, and graphical workflow system is integrated to represent real-time analysis progression. The BioVLAB-mCpG-SNP-EXPRESS system works in three stages. First, it processes and analyzes multi-omics data as input in the form of the raw data, i.e., FastQ files. Second, various integrated analyses such as methylation vs. gene expression and mutation vs. methylation are performed. Finally, the analysis result can be explored in a number of ways through a web interface for the multi-level, multi-perspective exploration. Multi-level interpretation can be done by either gene, gene set, pathway or network level and multi-perspective exploration can be explored from either gene expression, DNA methylation, sequence variation, or their relationship perspective. The utility of the system is demonstrated by performing analysis of phenotypically distinct 30 breast cancer cell line data set. BioVLAB-mCpG-SNP-EXPRESS is available at http://biohealth.snu.ac.kr/software/biovlab_mcpg_snp_express/.


Identification of circulating endorepellin LG3 fragment: Potential use as a serological biomarker for breast cancer.
Comparative proteome analysis was performed on the cultured media of human nontumor and malignant breast cell lines, Hs578Bst and Hs578T, respectively, in search of a serological biomarker(s) for breast cancer. Proteins in the conditioned media were separated by 2-D PAGE and then visualized by silver-staining. Eight proteins changed differentially by more than two-fold were identified by MALDI-TOF/TOF MS. Among the proteins identified, the terminal laminin-like globular (LG3) domain of endorepellin, which was recently reported as an antiangiogenesis factor, was decreased in the cancer cell line. We confirmed the bone morphogenic protein-1 (BMP-1) mediated cleavage site on the N-terminus of endorepellin LG3 fragment. This finding suggests that the LG3 fragment is specifically released by a BMP-1 driven limited proteolytic process. The protein was also detected in plasma by Western blot analysis and selected reaction monitoring (SRM). The plasma level of the endorepellin LG3 fragment was significantly lower in breast cancer patients compared to healthy donors (p = 0.017; n = 12). The LG3 protein concentration in the control plasma was measured at approximately 3.7 pmol/mL compared to 1.8 pmol/mL in plasma from the cancer patients. We suggest that these results support the potential use of the endorepellin LG3 fragment as a new serological biomarker for breast cancer.


No fault compensation in perinatal medicine in Japan-from results for 8 years.
No fault compensation in perinatal medicine has been established to provide aid to patients in the event of disability due to a medical accident during delivery and to strive to resolve disputes quickly that contributes to reduce medical malpractice suit in the field of perinatal medicine. Furthermore, this system is aimed to establish a mechanism that achieves higher quality of obstetric care by analyzing the causes of accidents. This system is operated by Japan Council for Quality Health Care and 99.9% of childbirth facilities in Japan registered with this compensation system. Compensation system has two major functions including compensation and cause analysis and recurrence prevention based on cases. Compensation eligibility is reviewed in the Review Committee at the organization in Japan Council for Quality Health Care and currently 1,717 cases are judged as eligible cases out of 2,250 since 2009. The cause of each eligible case is analyzed in the Cause Analysis Committee one by one. The Cause Analysis Committee has 7 independent subcommittee and each subcommittee members are consistent of obstetricians, neonatologists, pediatricians, midwives and lawyers. Original cause analysis report is sent to childbirth facility and patient's family. Questionnaire survey demonstrated that 73% of childbirth facilities and 65% of patient family were satisfied with the cause analysis report. The number of medical lawsuit in obstetrics and gynecology is significantly decreased compared to those in all medical departments since the compensation system was introduced suggesting that these cause analysis reports may contribute the decrease in medical lawsuits. The major purpose of the Recurrence Prevention Committee is striving to prevent future cases of cerebral palsy and to improve the quality of obstetric care. To accomplish the purpose, the committee routinely collect information from individual cases and analyze quantitatively and epidemiologically. Furthermore the committee choose subject from cases to discuss for future prevention and provide wide public disclosure of the information with recommendation. The committee issued 6 reports until now and these reports have been distributed to childbirth facilities, perinatal medicine-related scientific society and administrative agencies.


The impact of changes in Finland's health care system.
The most important reform in Finnish health care in the last decade occurred at the beginning of 1993 as part of a broader change to the entire state subsidy system. This reform reduced central government control and increased the freedom of the municipalities in the provision of health services. In addition, an unusually severe economic recession in the early 1990s affected Finnish health care. Currently, the most important economic decisions in the health care system are made by the 432 municipalities, which decide annually the amount of money to be devoted to health care as well as dividing resources in different sectors within the area of health. The changes in measures of productivity (based on activity-based measures in output) in the system are more closely associated with direct economic constraints (of municipalities) than with changes in financial incentive structures. Studies on equity in utilisation indicate that the Finnish health care system met the challenges of the 1991--1994 recession. Inequity in utilisation still prevails, however, and can be partly explained by the specific characteristics of the Finnish health care system. In future one needs to take more careful account of the rather unusual incentives that affect the behaviour of political decision-makers, providers and patients.


PBX3 is targeted by multiple miRNAs and is essential for liver tumour-initiating cells.
Tumour-initiating cells (TICs) are advocated to constitute the sustaining force to maintain and renew fully established malignancy; however, the molecular mechanisms responsible for these properties are elusive. We previously demonstrated that voltage-gated calcium channel α2δ1 subunit marks hepatocellular carcinoma (HCC) TICs. Here we confirm directly that α2δ1 is a HCC TIC surface marker, and identify let-7c, miR-200b, miR-222 and miR-424 as suppressors of α2δ1(+) HCC TICs. Interestingly, all the four miRNAs synergistically target PBX3, which is sufficient and necessary for the acquisition and maintenance of TIC properties. Moreover, PBX3 drives an essential transcriptional programme, activating the expression of genes critical for HCC TIC stemness including CACNA2D1, EpCAM, SOX2 and NOTCH3. In addition, the expression of CACNA2D1 and PBX3 mRNA is predictive of poor prognosis for HCC patients. Collectively, our study identifies an essential signalling pathway that controls the switch of HCC TIC phenotypes.


Differences in urinary excretion of steroid 21-deoxyketols by women pregnant with a normal or an anencephalic foetus.
During late pregnancy the urinary excretion of steroid 21-deoxyketols, which contain the CH3-CO-C(OH)less than side chain, was 1.16 +/- 0.50 mg/24 h (mean +/- SD n = 9). The mean excretion of these metabolites in 8 pregnancies with an anencephalic foetus showed a significant reduction (P less than 0.01, t-test) to 0.42 +/- 0.18 mg/24 h. Six untreated non-pregnant women and 7 taking oral contraceptives which contain oestrogen had steroid 21-deoxyketol excretions of 0.26 +/- 0.09 mg/24 h and 0.19 +/- 0.09 mg/24 h respectively. These results imply that the adrenal of the normal foetus makes a contribution to the secretion of those steroids which are ultimately excreted by the mother as 21-deoxyketols.