Current trauma team activation processes at Canadian trauma centres: A national survey.
Trauma team activation (TTA) allows the provision of specialized and timely care to improve outcomes for severely injured patients. Limited information is available on the current state of TTA in Canadian trauma centres (TC). Study objectives were to describe TTA processes, data and reports, along with the challenges and successes from a national perspective. A mixed-methods, cross-sectional survey was undertaken with Canadian trauma leadership, utilizing a total population sampling strategy. The questionnaire, containing 108-items, was administered online between February-April 2022, utilizing a modified Dillman technique. Descriptive statistics and thematic analyses were performed. Trauma leaders from 9 out of 10 provinces responded for a response rate of 68% (32/47). Two-thirds (67%) of respondents worked in adult TC; 63% in a level I center. A higher proportion of pediatric TC had a two-tiered TT response (60% pediatric; 35% adult). The most common criteria were neurologic compromise (100% one-level TTA) and hypotension (pediatric: 100% one-level, 100% tier 1; adult: 92% one-level, 86% tier 1). All one-level TTA included penetrating trauma criteria. One-third of respondents reported using TTA subgroup criteria for pediatric, pregnant, and/or geriatric patients. There was variability with disciplines responding to TTA, with largest, most comprehensive teams for tier 1. Two-thirds of TC review activation compliance (under/overtriage), while 55% focus on non-compliance and reasons for missed TTA. The most frequent challenges related to TTA practices were reliable data collection (60%) while successes included were the establishment of TTA guidelines to improve team compliance (33%) and RN initiated TTA. Some TTA practices were similar among Canadian TC, while others showed variability. Findings provide opportunities for improvement, including a two-tier system, geriatric-specific criteria, and RN initiated TTA, and could help establish national standards and best practices. Compliance with standards has the potential to improve Canadian TTA practices and patient outcomes.


[The repercussions of mental disease in the family. A study of the family members of psychotic patients].
Caregiving and related concepts such as family or caregiver burden have been widely discussed in recent literature. A descriptive, cross-sectional study was conducted in a sample (n = 80) of primary caregivers to severe mental patients (diagnosed schizophrenia, chronic psychosis or affective disorder). To describe the caregivers' and corresponding patients' sample, as well as the nature of existing relationships. Levels of caregiver burden and psychological distress were evaluated and a preliminary analysis conducted on objective and subjective burden correlation. Involvement Evaluation Questionnaire (IEQ), General Health Questionnaire (GHQ) and a semi-structured interview designed for this purpose. The extent of burden was found significant in this sample, being unevenly distributed in its components (e.g. tension, worrying or financial burden). Forty-one percent of caregivers experienced minor psychiatric morbidity. Measures of objective and subjective burden were moderately correlated. The same applied to magnitude of associations between each of them and distress parameters. Higher levels of burden were found in groups of caregivers to more severely ill or disabled patients, while diagnosis was not important for this purpose. Most relatives remain highly involved. These results suggest the need to implement specific interventions addressing psychotic patients as well as their families.


Tongue viability after bilateral lingual artery ligation and surgery for recurrent tongue-base cancer.
A 73-year-old man came to us with a second primary tumor of the tongue base. He had undergone a total laryngectomy, neck dissections, and radiation for squamous cell carcinoma of the glottis 25 years earlier. Surgery was performed, including a subtotal tongue-base resection and pectoralis myocutaneous flap reconstruction. Intraoperative findings confirmed tumor involvement of the left lingual artery and hypoglossal nerve, both of which were sacrificed. Dissection revealed an intact but apparently thrombosed right lingual artery, and this was verified by intraoperative Doppler evaluation. Postoperatively, the preserved oral tongue remained well perfused and viable, and tongue mobility was good. In patients who have undergone previous head and neck cancer resections, collateral blood supply to the tongue might develop if the flow of one or both of the lingual arteries is interrupted. This possibility can be fully evaluated by preoperative angiography, which might offer these patients the potential for tongue preservation.


Mycophenolate mofetil for scleroderma-related interstitial lung disease: A real world experience.
Interstitial lung disease (ILD) remains the number one cause of mortality in scleroderma (SSc). Our goal was to determine the effectiveness of mycophenolate mofetil (MMF) in treating SSc-ILD in a retrospective study. A retrospective, computer-assisted search was performed to identify patients with SSc-ILD treated with MMF from 1997 through 2014. We used a novel software tool, Computer-Aided Lung Informatics for Pathology Evaluation and Rating (CALIPER), to quantify parenchymal lung abnormalities on high-resolution computed tomography. Lung function was evaluated at baseline, 6, 12, and 24 months of MMF therapy. We identified 46 patients (28 females) with SSc-ILD (mean age at diagnosis 55 y) treated with MMF for at least 1 year (majority on 2 gm/day). Twenty-one patients (45.7%) stopped using MMF during the follow up period after the first 12 months, and they took MMF for a median of 2.12 years (range, 0.91-8.93 years). Only 4 discontinued MMF because of disease progression. Compared to baseline, the mean percentage change in forced vital capacity (95% CI) at 6, 12, and 24 months, respectively, was 1.01% (-2.38%-4.39%) (n = 26), 2.06% (-1.09%-5.22%) (n = 31), and -0.07% (-3.31%-3.17%) (n = 30), and the mean percentage change in ILD as measured by CALIPER (95% CI) was -5.40% (-18.62%-7.83%) (n = 18), -1.51% (-14.69%-11.68%) (n = 17), and -8.35% (-20.71%-4.02%) (n = 22).The mean right ventricular systolic pressure (RVSP) remained stable over the study period. MMF is well tolerated and slows the rate of decline in lung function in SSc-ILD patients, even at doses lower at 3 g/day.


Diagnostic techniques used for ectopic pregnancy.
Though all diagnostic techniques used for ectopic pregnancy yield false-negative and false-positive results, prospective serial studies using hormonal and sonographic testing are quite accurate when appropriately applied.


Fasciola hepatica glycoconjugates immuneregulate dendritic cells through the Dendritic Cell-Specific Intercellular adhesion molecule-3-Grabbing Non-integrin inducing T cell anergy.
Dendritic cell-specific ICAM-3 grabbing non-integrin (DC-SIGN) expressed on a variety of DCs, is a C-type lectin receptor that recognizes glycans on a diverse range of pathogens, including parasites. The interaction of DC-SIGN with pathogens triggers specific signaling events that modulate DC-maturation and activity and regulate T-cell activation by DCs. In this work we evaluate whether F. hepatica glycans can immune modulate DCs via DC-SIGN. We demonstrate that DC-SIGN interacts with F. hepatica glycoconjugates through mannose and fucose residues. We also show that mannose is present in high-mannose structures, hybrid and trimannosyl N-glycans with terminal GlcNAc. Furthermore, we demonstrate that F. hepatica glycans induce DC-SIGN triggering leading to a strong production of TLR-induced IL-10 and IL-27p28. In addition, parasite glycans induced regulatory DCs via DC-SIGN that decrease allogeneic T cell proliferation, via the induction of anergic/regulatory T cells, highlighting the role of DC-SIGN in the regulation of innate and adaptive immune responses by F. hepatica. Our data confirm the immunomodulatory properties of DC-SIGN triggered by pathogen-derived glycans and contribute to the identification of immunomodulatory glyans of helminths that might eventually be useful for the design of vaccines against fasciolosis.


The evaluation of topical administration of Bellis perennis fraction on circular excision wound healing in Wistar albino rats.
Bellis perennis L. (Asteraceae) has been used traditionally in the treatment of bruises, broken bones, and wounds by European people. To investigate the wound healing activity of B. perennis flowers in Wistar albino rats. Dried B. perennis flowers were extracted with ethanol, then fractioned with n-butanol and an oinment was prepared. Twelve male adult Wistar rats were used. Six wounds were created for each animal by using circular excision wound model. The first two wounds were treated topically with HOTBp (hydrophilic ointment treatment containing n-butanol fraction). The second two wounds were control group and not treated with anything. The third two wounds were treated only with HOT (hydrophilic ointment treatment without n-butanol fraction). Treatments were applied once a day and lasted for 30 days. Wound samples were excised on days 5(th), 10(th) and 30(th). The percentage of wound healing was calculated by Walker's formula after measurement of the wound area and the tissue samples were examined histopathologically. The percentages of wound closure (HOTBp: 100%; HOT: 85% and control: 87%) and histopathological observations showed that there were statistically significant differences between HOTBp, HOT and control groups (p < 0.05) at 30(th) day. Topically administered ointment prepared from the n-butanol fraction of B. perennis flowers has a wound healing potential without scar formation in circular excision wound model in rats. Thus, traditional usage of wound healing activity of B. perennis was scientifically verified for the first time.


EPAS1 trans-activation during hypoxia requires p42/p44 MAPK.
Hypoxia is a common environmental stress that regulates gene expression and cell function. A number of hypoxia-regulated transcription factors have been identified and have been shown to play critical roles in mediating cellular responses to hypoxia. One of these is the endothelial PAS-domain protein 1 (EPAS1/HIF2-alpha/HLF/HRF). This protein is 48% homologous to hypoxia-inducible factor 1-alpha (HIF1-alpha). To date, virtually nothing is known about the signaling pathways that lead to either EPAS1 or HIF1-alpha activation. Here we show that EPAS1 is phosphorylated when PC12 cells are exposed to hypoxia and that p42/p44 MAPK is a critical mediator of EPAS1 activation. Pretreatment of PC12 cells with the MEK inhibitor, PD98059, completely blocked hypoxia-induced trans-activation of a hypoxia response element (HRE) reporter gene by transfected EPAS1. Likewise, expression of a constitutively active MEK1 mimicked the effects of hypoxia on HRE reporter gene expression. However, pretreatment with PD98059 had no effect on EPAS1 phosphorylation during hypoxia, suggesting that MAPK targets other proteins that are critical for the trans-activation of EPAS1. We further show that hypoxia-induced trans-activation of EPAS1 is independent of Ras. Finally, pretreatment with calmodulin antagonists nearly completely blocked both the hypoxia-induced phosphorylation of MAPK and the EPAS1 trans-activation of HRE-Luc. These results demonstrate that the MAPK pathway is a critical mediator of EPAS1 activation and that activation of MAPK and EPAS1 occurs through a calmodulin-sensitive pathway and not through the GTPase, Ras. These results are the first to identify a specific signaling pathway involved in EPAS1 activation.


Patients' Perceptions of Integrated Care Among Medicare Beneficiaries by Level of Need for Health Services.
Requirements for integrating care across providers, settings, and over time increase with patients' needs. Health care providers' ability to offer care that patients experience as integrated may vary among patients with different levels of need. We explore the variation in patients' perceptions of integrated care among Medicare beneficiaries based on the beneficiary's level of need using ordinary least square regression for each of four high-need groups: beneficiaries (a) with complex chronic conditions, (b) with frailties, (c) below 65 with disability, and (d) with any (of the first three) high needs. We control for beneficiary demographics and other factors affecting integrated care, and we conduct sensitivity analyses controlling for multiple individual chronic conditions. We find significant positive associations with level of need for provider support for self-directed care and medication and home health management. Controlling for multiple individual chronic conditions reduces effect sizes and number of significant relationships.


Physical Functional Capacity and C-Reactive Protein in Schizophrenia.
Schizophrenia is a severe, debilitating mental disorder that affects both the physical health and the functional capacity of patients, causing great impairment throughout the life course. Although physical and cognitive impairments may represent different expressions of a single systemic inflammatory process, little is known about the relationship between motor function and schizophrenia. To evaluate physical functional capacity in patients with schizophrenia and ascertain whether it correlates with markers of inflammation, disease severity, and pharmacotherapy. Cross-sectional study using a convenience sampling strategy. Forty patients with stable schizophrenia, undergoing treatment, were recruited from the Outpatient Program of Hospital de Clínicas de Porto Alegre, University Hospital linked to Public Health System. Physical functional capacity was assessed by the 6-min walk test (6MWT), and inflammatory markers were measured by C-reactive protein (CRP) and Von Willebrand factor. Mean functional capacity and clinical variables differed among patients and Brazilian population regarding heart rate (p = 0.004), diastolic (p = 0.001) and systolic (p < 0.001) blood pressure, respiratory rate (p < 0.001), CRP (p = 0.015), Borg Scale of Perceived Exertion scores (BSPE) (p < 0.001), and 6MWT both in men (p < 0.001) and women (p = 0.024). Additionally, 6MWT and dyspnea in BSPE were positively associated with CRP (r = -0.369, p = 0.019) and (r = -0.376, p = 0.017) and (r = 0.354, p = 0.025 and r = 0.535, p < 0.001, respectively). The present study detected significant association between measures of functional impairment and markers of inflammation, especially elevated CRP in a group of stable outpatients with DSM-IV and ICD10 diagnosis of schizophrenia. Possible explanations for the associations could be linked to continued use of antipsychotics, although underlying neuroinflammatory mechanisms directly related to illness (schizophrenia) could not be ruled out. The findings of this study expand evidences of neuroinflammation to systemic inflammation in schizophrenia linking it to alterations of physical functional capacity and point to the need of additional studies exploring general inflammation and novel therapeutic interventions.


Updating the Vibrio clades defined by multilocus sequence phylogeny: proposal of eight new clades, and the description of Vibrio tritonius sp. nov.
To date 142 species have been described in the Vibrionaceae family of bacteria, classified into seven genera; Aliivibrio, Echinimonas, Enterovibrio, Grimontia, Photobacterium, Salinivibrio and Vibrio. As vibrios are widespread in marine environments and show versatile metabolisms and ecologies, these bacteria are recognized as one of the most diverse and important marine heterotrophic bacterial groups for elucidating the correlation between genome evolution and ecological adaptation. However, on the basis of 16S rRNA gene phylogeny, we could not find any robust monophyletic lineages in any of the known genera. We needed further attempts to reconstruct their evolutionary history based on multilocus sequence analysis (MLSA) and/or genome wide taxonomy of all the recognized species groups. In our previous report in 2007, we conducted the first broad multilocus sequence analysis (MLSA) to infer the evolutionary history of vibrios using nine housekeeping genes (the 16S rRNA gene, gapA, gyrB, ftsZ, mreB, pyrH, recA, rpoA, and topA), and we proposed 14 distinct clades in 58 species of Vibrionaceae. Due to the difficulty of designing universal primers that can amplify the genes for MLSA in every Vibrionaceae species, some clades had yet to be defined. In this study, we present a better picture of an updated molecular phylogeny for 86 described vibrio species and 10 genome sequenced Vibrionaceae strains, using 8 housekeeping gene sequences. This new study places special emphasis on (1) eight newly identified clades (Damselae, Mediterranei, Pectenicida, Phosphoreum, Profundum, Porteresiae, Rosenbergii, and Rumoiensis); (2) clades amended since the 2007 proposal with recently described new species; (3) orphan clades of genomospecies F6 and F10; (4) phylogenetic positions defined in 3 genome-sequenced strains (N418, EX25, and EJY3); and (5) description of V. tritonius sp. nov., which is a member of the "Porteresiae" clade.


15-(p[75Br]bromophenyl)pentadecanoic acid: pharmacokinetics and potential as heart agent.
Preparation, quality control, and pharmacokinetics of 15-(p-[75Br]bromophenyl)pentadecanoic acid (BPPA) were studied, with particular emphasis on the fate of the label and the usefulness of BPPA as a heart agent. BPPA labeled with the positron emitter Br-75 (T 1/2 = 98 min) was prepared in 55% radiochemical yield with a specific activity of greater than or equal to 1000 Ci/mmol. While the uptake of BPPA in the heart muscle is as fast and efficient as that of aliphatic omega-halofatty acids, its elimination is delayed, owing mainly to an inhibited beta oxidation and the formation of lipophilic catabolites. The blood activity and its time course are identical to those of omega-halofatty acids, but no free bromide appears in any body fluid or organ. The complex pharmacokinetic behavior of stable lipophilic catabolites prevents a quantitative background correction. In contrast to aliphatic omega-halofatty acids, a quantitative evaluation of myocardial metabolism is not possible. Br-75 BPPA, however, is well suited for positron emission tomography of the heart.


Segmenting/Pre-Processing Data from Bone Screw Thread-Stripping Tests.
Bone screws must be appropriately tightened to achieve optimal patient outcomes. If over-torqued, the threads formed in the bone may break, compromising the strength of the fixation; and, if under-torqued, the screw may loosen over time, compromising the stability. Previous work has proposed a model-based system to automatically determine the optimal insertion torque. This system consists of a reverse-modelling step to determine strength, and a forward modelling step to determine maximum torque. These have previously been tested in isolation, however future work must test the combined system. To do so, the data must be segmented and pre-processed. This was done based on specific features of the recorded data. The methodology was tested on 50 screw-insertion data sets across 5 different materials. With the parameters used, all data sets were correctly segmented. This will form a basis for the further processing of the data and validating the combined systemClinical relevance: The system for torque limit determination must be tested in its entirety to properly asses its performance. This paper discusses some of the steps required to pre-process the data to make this assessment. If successful, this system may improve patient outcomes in orthopaedic surgery.


Waiting to be born: the ethical implications of the generation of "NUBorn" and "NUAge" mice from pre-pubertal ovarian tissue.
Oncofertility is one of the 9 NIH Roadmap Initiatives, federal grants intended to explore previously intractable questions, and it describes a new field that exists in the liminal space between cancer treatment and its sequelae, IVF clinics and their yearning, and basic research in cell growth, biomaterials, and reproductive science and its tempting promises. Cancer diagnoses, which were once thought universally fatal, now often entail management of a chronic disease. Yet the therapies are rigorous, must start immediately, and in many cases result in premature failure of the body's reproductive ability. In women, this loss is especially poignant; unlike the routine storage of sperm, which is done in men and boys facing similar treatment decisions, freezing oocytes in anticipation of fertility loss is not possible in most cases, and creating an embryo within days of diagnosis raises significant moral, social and medical challenges. Oncofertility is the study of how to harvest ovarian tissue in women facing cancer to preserve their gametes for future use with IVF, thus allowing the decisions about childbearing to be deferred and reproductive choices to be preserved. The research endeavor uses the capacity of the ovarian follicle to produce eggs in vitro. Developing the human follicle to ovulate successfully outside the body is scientifically difficult and ethically challenging. Infertility is linked to long-standing religious and moral traditions, and is intertwined with deeply contentious social narratives about women, families, illness and birth. Is the research morally permissible? Perhaps imperative if understood as a repair from iatrogenic harms? How are considerations of justice central to the work? How will vulnerable subjects be protected? What are the moral implications of the work for women, children and families? What are the implications for society if women could store ovarian tissue as a way of stopping the biological clock? What are the moral possibilities and challenges if eggs can be produced in large quantities from a stored ovarian tissue?


Long-term health outcomes of childhood sexual abuse and peer sexual contact among an urban sample of behaviourally bisexual Latino men.
While previous research indicates high rates of childhood sexual abuse among Latino men who have sex with men, few studies have examined the long-term health outcomes of childhood sexual abuse specifically among behaviourally bisexual Latino men. In a sample of 148 behaviourally bisexual Latino men in New York City, we examined associations between childhood sexual abuse and multiple dimensions of adult health: sexual risk behaviours; sexually transmitted infections incidence; polydrug use; depressive symptoms; and perceived stress. We compared outcomes between those with histories of childhood sexual abuse, those reporting peer sexual contact prior to age 13 and those with no sexual contact prior to age 13. Over one-fifth (22.3%) reported a history of childhood sexual abuse, which was significantly associated with engaging in receptive condomless anal intercourse (aOR = 3.59, p < .01, SE = 2.0), high perceived stress (aOR = 2.48, p < .06, SE = 1.13) and clinically significant depressive symptoms (aOR = 2.7, p < .05, SE = 1.25). Across all variables, peer sexual contact did not impact these outcomes, underscoring a key distinction between abusive and non-abusive early sexual experiences. We recommend that sexual abuse prevention policies and programmes better engage Latino youth, and that practitioners serving this population across diverse areas of practice incorporate childhood sexual abuse screening and culturally appropriate treatment and care into practice.


Comparison of video laryngoscopy and direct laryngoscopy in a critical care transport service.
We evaluated video laryngoscopy (VL) (C-MAC, Karl Storz, Tuttlingen, Germany) for use in a critical care transport system. We hypothesized that the total number of airway attempts would decrease when using a video laryngoscope versus use of direct laryngoscopy (DL). We performed a nonrandomized group-controlled trial where six aircraft were outfitted with VL and the remainder utilized DL responding to a mix of scene runs and interfacility transports. Our primary outcome measure was the number of intubation attempts. We also compared the first-pass success (FPS) rates, laryngoscopic grades, and frequencies of rescue device use (including utilization of surgical airways) between VL and DL. Crews intubated 348 patients with VL and 510 with DL. Successful endotracheal intubation within three attempts occurred 97.6% (confidence interval [CI] 96.5-98.6) of the time. The FPS rate was 85.8% (CI 83.4-88.1). In this cohort of patients, VL did not differ from DL with respect to total number of airway attempts (1.17 [CI 1.11-1.22] vs. 1.16 [CI 1.12-1.20]), FPS rate (85.6% [CI 82-89%] vs. 86.1% [CI 83-89]), or use of rescue airways (2.6% vs. 2.2%). The laryngoscopic view was superior in the VL group relative to the DL group (median Cormack-Lehane grade 1 [interquartile range (IQR) 1, 2] vs. 2 [IQR 1, 2]). VL using the C-MAC video laryngoscope did not reduce the total number of airway attempts or improve intubation compared with DL in a system of highly trained providers.


Injectable, scalable 3D tissue-engineered model of marrow hematopoiesis.
Modeling the interaction between the supportive stroma and the hematopoietic stem and progenitor cells (HSPC) is of high interest in the regeneration of the bone marrow niche in blood disorders. In this work, we present an injectable co-culture system to study this interaction in a coherent in vitro culture and in vivo transplantation model. We assemble a 3D hematopoietic niche in vitro by co-culture of supportive OP9 mesenchymal cells and HSPCs in porous, chemically defined collagen-coated carboxymethylcellulose microscaffolds (CCMs). Flow cytometry and hematopoietic colony forming assays demonstrate the stromal supportive capacity for in vitro hematopoiesis in the absence of exogenous cytokines. After in vitro culture, we recover a paste-like living injectable niche biomaterial from CCM co-cultures by controlled, partial dehydration. Cell viability and the association between stroma and HSPCs are maintained in this process. After subcutaneous injection of this living artificial niche in vivo, we find maintenance of stromal and hematopoietic populations over 12 weeks in immunodeficient mice. Indeed, vascularization is enhanced in the presence of HSPCs. Our approach provides a minimalistic, scalable, biomimetic in vitro model of hematopoiesis in a microcarrier format that preserves the HSPC progenitor function, while being injectable in vivo without disrupting the cell-cell interactions established in vitro.


Pearl millet [Pennisetum glaucum (L.) R. Br.] consensus linkage map constructed using four RIL mapping populations and newly developed EST-SSRs.
Pearl millet [Pennisetum glaucum (L.) R. Br.] is a widely cultivated drought- and high-temperature tolerant C4 cereal grown under dryland, rainfed and irrigated conditions in drought-prone regions of the tropics and sub-tropics of Africa, South Asia and the Americas. It is considered an orphan crop with relatively few genomic and genetic resources. This study was undertaken to increase the EST-based microsatellite marker and genetic resources for this crop to facilitate marker-assisted breeding. Newly developed EST-SSR markers (99), along with previously mapped EST-SSR (17), genomic SSR (53) and STS (2) markers, were used to construct linkage maps of four F7 recombinant inbred populations (RIP) based on crosses ICMB 841-P3 × 863B-P2 (RIP A), H 77/833-2 × PRLT 2/89-33 (RIP B), 81B-P6 × ICMP 451-P8 (RIP C) and PT 732B-P2 × P1449-2-P1 (RIP D). Mapped loci numbers were greatest for RIP A (104), followed by RIP B (78), RIP C (64) and RIP D (59). Total map lengths (Haldane) were 615 cM, 690 cM, 428 cM and 276 cM, respectively. A total of 176 loci detected by 171 primer pairs were mapped among the four crosses. A consensus map of 174 loci (899 cM) detected by 169 primer pairs was constructed using MergeMap to integrate the individual linkage maps. Locus order in the consensus map was well conserved for nearly all linkage groups. Eighty-nine EST-SSR marker loci from this consensus map had significant BLAST hits (top hits with e-value ≤ 1E-10) on the genome sequences of rice, foxtail millet, sorghum, maize and Brachypodium with 35, 88, 58, 48 and 38 loci, respectively. The consensus map developed in the present study contains the largest set of mapped SSRs reported to date for pearl millet, and represents a major consolidation of existing pearl millet genetic mapping information. This study increased numbers of mapped pearl millet SSR markers by >50%, filling important gaps in previously published SSR-based linkage maps for this species and will greatly facilitate SSR-based QTL mapping and applied marker-assisted selection programs.


HLA-DR antigens in primary biliary cirrhosis: lack of association.
A study of HLA-DR antigen in 75 patients with primary biliary cirrhosis has been carried out in order to test the hypothesis that genetic factors related to genes controlling immune responses might be important in the pathogenesis of primary biliary cirrhosis. The frequencies of HLA-DR locus antigens was not significantly different from those in 200 normal controls, nor were those of tissue antigens on the A and B loci. No HLA-DR antigen was significantly associated with the appearance of granulomata on liver biopsy (possibly good prognosis) or with raised serum bilirubin (possibly bad prognosis); nor was there any association between any HLA-DR antigen and adverse reactions to D-penicillamine treatment in 17 patients with such adverse reactions. It is concluded that genetic traits related to HLA antigens studied are probably not important in the aetiology of the disease.


Investigation of electron trajectories of an x-ray tube in magnetic fields of MR scanners.
A hybrid x-ray/MR system combining an x-ray fluoroscopic system and an open-bore magnetic resonance (MR) system offers advantages from both powerful imaging modalities and thus can benefit numerous image-guided interventional procedures. In our hybrid system configurations, the x-ray tube and detector are placed in the MR magnet and therefore experience a strong magnetic field. The electron beam inside the x-ray tube can be deflected by a misaligned magnetic field, which may damage the tube. Understanding the deflection process is crucial to predicting the electron beam deflection and avoiding potential damage to the x-ray tube. For this purpose, the motion of an electron in combined electric (E) and magnetic (B) fields was analyzed theoretically to provide general solutions that can be applied to different geometries. For two specific cases, a slightly misaligned strong field and a perpendicular weak field, computer simulations were performed with a finite-element method program. In addition, experiments were conducted using an open MRI magnet and an inserted electromagnet to quantitatively verify the relationship between the deflections and the field misalignment. In a strong (B >> E/c; c: speed of light) and slightly misaligned magnetic field, the deflection in the plane of E and B caused by electrons following the magnetic field lines is the dominant component compared to the deflection in the E X B direction due to the drift of electrons. In a weak magnetic field (B < or = E/c), the main deflection is in the E x B direction and is caused by the perpendicular component of the magnetic field.


Optimization of initial prostate biopsy in clinical practice: sampling, labeling and specimen processing.
An optimal prostate biopsy in clinical practice is based on a balance among adequate detection of clinically significant prostate cancers (sensitivity), assuredness regarding the accuracy of negative sampling (negative predictive value), limited detection of clinically insignificant cancers and good concordance with whole gland surgical pathology results to allow accurate risk stratification and disease localization for treatment selection. Inherent within this optimization is variation of the core number, location, labeling and processing for pathological evaluation. To date, there is no consensus in this regard. The purpose of this review is to 1) define the optimal number and location of biopsy cores during primary prostate biopsy among men with suspected prostate cancer, 2) define the optimal method of labeling prostate biopsy cores for pathological processing which will provide relevant and necessary clinical information for all potential clinical scenarios, and 3) determine the maximal number of prostate biopsy cores allowable within a specimen jar which would not preclude accurate histological evaluation of the tissue. A bibliographic search using PubMed® covering the period up to July 2012 yielded approximately 550 articles. Articles were reviewed and categorized based on which of the 3 objectives of this review was addressed. Data were extracted, analyzed and summarized. Recommendations are provided based on this literature review and our clinical experience. The use of 10 to 12-core extended sampling protocols increases cancer detection rates compared to traditional sextant sampling methods and reduces the likelihood of repeat biopsy by increasing negative predictive value, ultimately allowing more accurate risk stratification without increasing the likelihood of detecting insignificant cancers. As the number of cores increases above 12, the increase in diagnostic yield becomes marginal. Only limited evidence supports the use of initial biopsy schemes involving more than 12 cores or saturation. Apical and laterally directed sampling of the peripheral zone increases cancer detection rate, reduces the need for repeat biopsies and predicts pathological features on prostatectomy while transition zone biopsies do not. There are little data to suggest that knowing the exact site of an individual positive biopsy core provides meaningful clinical information. However, determining laterality of cancer on biopsy may be helpful for predicting sites of extracapsular extension and therapeutic planning. Placement of multiple biopsy cores in a single container (greater than 2) appears to compromise pathological evaluation, which can reduce cancer detection rate and increase the likelihood of equivocal diagnoses. A 12-core systematic biopsy that incorporates apical and far-lateral cores in the template distribution allows maximal cancer detection, avoids repeat biopsy, and provides information adequate for identifying men who need therapy and planning that therapy while minimizing the detection of occult, indolent prostate cancers. This literature review does not provide compelling evidence that individual site specific labeling of cores benefits clinical decision making regarding the management of prostate cancer. Based on the available literature, we recommend packaging no more than 2 cores in each jar to avoid reduction of the cancer detection rate through inadequate tissue sampling.


Association of Smoking Status with Efficacy of First-line Immune Checkpoint Inhibitors in Advanced Non-small Cell Lung Cancers: A Systematic Review and Meta-analysis.
Background: Although smoking status has potential as a biomarker for immune checkpoint blockade in advanced non-small cell lung cancer (NSCLC), its clinical significance remains obscure. This meta-analysis aims to assess the impact of the smoking status on the efficacy of first-line immunotherapy and to find better treatment in never-smoker and ever-smoker patients. Methods: We searched the MEDLINE, EMBASE, and Cochrane database for trials comparing immunotherapy with conventional chemotherapy as front-line treatment for advanced NSCLC. Random-effects models were used to pool estimates of hazard ratios (HRs) for overall survival with 95% confidence intervals (CIs). Predefined subgroup analysis was performed to investigate the difference in the efficacy between the single checkpoint blockade and checkpoint inhibitor plus chemotherapy combination in the never-smokers and current/former smokers. Results: Twelve trials involving 6,446 patients were included in the analysis. A statistically significant overall survival benefit over conventional chemotherapy was found for both checkpoint inhibitor monotherapy (HR, 0.71; 95% CI, 0.59-0.85) and checkpoint inhibitor plus chemotherapy (HR, 0.75; 95% CI, 0.63-0.90) in the current/former smoker group. There was no subgroup difference between monotherapy and combination treatment (p=0.67). However, there was an inconsistent survival outcome in the never-smoker group; checkpoint blockade monotherapy did not show significantly better efficacy than chemotherapy alone (HR, 1.05; 95% CI, 0.81-1.37), but combination treatment showed an overall survival benefit (HR, 0.64; 95% CI, 0.43-0.94). A significant subgroup difference existed between monotherapy and combination therapy (p=0.04). Similarly, there was a significant difference in efficacy of monotherapy between the current/former smoker and never-smoker group (p=0.01), but the efficacy of the combination treatment was comparable between the two groups (p=0.45). Conclusion: Smoking status, which is easily available information, could be used as a guide in clinical practice to choose better treatment in the front-line setting for advanced NSCLC patients.


Detecting subtle yet fast skeletal muscle contractions with ultrasoft and durable graphene-based cellular materials.
Human bodily movements are primarily controlled by the contractions of skeletal muscles. Unlike joint or skeletal movements that are generally performed in the large displacement range, the contractions of the skeletal muscles that underpin these movements are subtle in intensity yet high in frequency. This subtlety of movement makes it a formidable challenge to develop wearable and durable soft materials to electrically monitor such motions with high fidelity for the purpose of, for example, muscle/neuromuscular disease diagnosis. Here we report that an intrinsically fragile ultralow-density graphene-based cellular monolith sandwiched between silicone rubbers can exhibit a highly effective stress and strain transfer mechanism at its interface with the rubber, with a remarkable improvement in stretchability (>100%). In particular, this hybrid also exhibits a highly sensitive, broadband-frequency electrical response (up to 180 Hz) for a wide range of strains. By correlating the mechanical signal of muscle movements obtained from this hybrid material with electromyography, we demonstrate that the strain sensor based on this hybrid material may provide a new, soft and wearable mechanomyography approach for real-time monitoring of complex neuromuscular-skeletal interactions in a broad range of healthcare and human-machine interface applications. This work also provides a new architecture-enabled functional soft material platform for wearable electronics.


[Primary penoscrotal extra-mammary Paget's disease: analysis of 22 cases].
To investigate the clinical characteristics, treatment and prognosis of primary penoscrotal extra-mammary Paget's disease (PSPD). The clinical and pathological data of 22 cases of primary PSPD were retrospectively reviewed. Survival rate of the overall patients, the invasive patients, and the patients with positive surgical margin or negative surgical margin were analyzed with Kaplan-Meier survival curve method. Among all the patients with primary PSPD, the median age of onset was 64.5 (39-84) years, the median time of disease duration was 40 (2-300) months, and the median long diameter of lesion was 4.75 (1-10) cm. In the study, 12 patients (54.5%) were in pathological stage A1, 6 patients (27.3%) were in pathological stage A2, and 4 patients (18.2%) were in pathological stage B. Scrotum and penile of most patients (n=12, 54.5%) were involved, 5 patients (22.7%) were scrotum only, and 5 patients (22.7%) were penile only. Erythema (n=18, 81.8%), itchiness (n=16, 72.7%), ulcerate (n=12, 54.5%), exudation (n=11, 50.0%), and pain (n=4, 18.2%) were the major manifestations. All the patients with primary PSPD were treated with wide surgical excision. The rate of invasive patients was 77.3% (n=17). Of them, 6 patients had positive surgical margin. The surgical margins of non-invasive patients were all negative. Twelve patients exhibited local recurrence or/and metastases, and the status of surgical margins (P=0.015) and the depth of invasion (P=0.010) were important risk factors. Inguinal lymph nodes were generally involved. The difference of the delay of diagnosis between metastasis and non-metastasis was significant (P=0.040). The 5-year survival rates of the overall and invasive patients were 33.7% and 27.9%, respectively. The patients with positive surgical margin had poor prognosis. Primary PSPD, with the characteristics of long duration, high invasive tendency and high incidence of local recurrence or metastases, generally occurs in the elderly. Surgery should be performed at first. The delay of diagnosis, positive surgical margins and the involvement of inguinal lymph node are important risk factors. Biopsy, frozen section and inguinal lymph node biopsy (ILNB) can standardize diagnosis and treatment.


Structure-based design of a novel series of azetidine inhibitors of the hepatitis C virus NS3/4A serine protease.
Structural homology between thrombin inhibitors and the early tetrapeptide HCV protease inhibitor led to the bioisosteric replacement of the P2 proline by a 2,4-disubstituted azetidine within the macrocyclic β-strand mimic. Molecular modeling guided the design of the series. This approach was validated by the excellent activity and selectivity in biochemical and cell based assays of this novel series and confirmed by the co-crystal structure of the inhibitor with the NS3/4A protein (PDB code: 4TYD).


Transcription regulation of the Escherichia coli pcnB gene coding for poly(A) polymerase I: roles of ppGpp, DksA and sigma factors.
Poly(A) polymerase I (PAP I), encoded by the pcnB gene, is a major enzyme responsible for RNA polyadenylation in Escherichia coli, a process involved in the global control of gene expression in this bacterium through influencing the rate of transcript degradation. Recent studies have suggested a complicated regulation of pcnB expression, including a complex promoter region, a control at the level of translation initiation and dependence on bacterial growth rate. In this report, studies on transcription regulation of the pcnB gene are described. Results of in vivo and in vitro experiments indicated that (a) there are three σ(70)-dependent (p1, pB, and p2) and two σ(S)-dependent (pS1 and pS2) promoters of the pcnB gene, (b) guanosine tetraphosphate (ppGpp) and DksA directly inhibit transcription from pB, pS1 and pS2, and (c) pB activity is drastically impaired at the stationary phase of growth. These results indicate that regulation of the pcnB gene transcription is a complex process, which involves several factors acting to ensure precise control of PAP I production. Moreover, inhibition of activities of pS1 and pS2 by ppGpp and DksA suggests that regulation of transcription from promoters requiring alternative σ factors by these effectors of the stringent response might occur according to both passive and active models.


Poly(vinyl alcohol) and Functionalized Ionic Liquid-Based Smart Hydrogels for Doxorubicin Release.
Limitations associated with the traditional cancer therapies prompt the scientific community to develop effective, safer, smarter, and targeted drug carriers that improve the efficiency of the drug carrier, reduce the adverse effects of the drug on the healthy cells, and help in preventing the cancer recurrences. This research aims to design a stimuli-responsive, self-healable, adhesive, and injectable polymeric hydrogel with an ester-functionalized ionic liquid as one of the additives to improve the efficiency of the anticancer drug in encapsulation and localized delivery. The designed polymeric hydrogel responds to intracellular biological stimuli (e.g., acidic pH of cancerous cells and temperature), changes the morphology through changing the shape and size of the gelator within the hydrogel matrix, and releases encapsulated doxorubicin (DOX) at the tumor site efficiently. Molecular interactions, gel morphology, and mechanical strength of the hydrogel were characterized through various analytical techniques, including small-angle neutron scattering. Adhesive properties of the polymeric hydrogel were measured by lap-shear strength tests and the biocompatibility and cellular drug uptake study on human breast cancer (MCF-7) and human cervical carcinoma cells (HeLa). The in vitro cytotoxicity and drug release study showed that the hybrid hydrogel is more effective at killing the cancerous cells, and the targeted release of DOX occurred at intracellular acidic pH. The polymeric hydrogel provides an efficient therapeutic approach for the encapsulation and release of the drug. Overall, the study offers a proof of concept to test the feasibility of the hydrogel system whether the hydrogel formulation helped or hindered the total cellular DOX trafficking.


Glutamatergic calcium responses in the developing lateral superior olive: receptor types and their specific activation by synaptic activity patterns.
The lateral superior olive (LSO) is a binaural auditory brain stem nucleus that plays a central role in sound localization. Survival and maturation of developing LSO neurons critically depend on intracellular calcium signaling. Here we investigated the mechanisms by which glutamatergic afferents from the cochlear nucleus increase intracellular calcium concentration in LSO neurons. Using fura-2 calcium imaging in slices prepared from neonatal mice, we found that cochlear nucleus afferents can activate all major classes of ionotropic and metabotropic glutamate receptors, each of which contributes to an increase in intracellular calcium. The specific activation of different glutamate receptor classes was dependent on response amplitudes and afferent stimulus patterns. Low-amplitude responses elicited by single stimuli were entirely mediated by calcium-impermeable AMPA/kainate receptors that activated voltage-gated calcium channels. Larger-amplitude responses elicited by either single stimuli or stimulus trains resulted in additional calcium influx through N-methyl-d-aspartate receptors. Finally, high-frequency stimulation also recruited group I and group II metabotropic glutamate receptors, both of which mobilized intracellular calcium. This calcium release in turn activated a strong influx of extracellular calcium through a membrane calcium channel that is distinct from voltage-gated calcium channels. Together, these results indicate that before hearing onset, distinct patterns of afferent activity generate qualitatively distinct types of calcium responses, which likely serve in guiding different aspects of LSO development.


Cellular vehicles for cancer gene therapy: current status and future potential.
Cancer is a difficult target for any therapeutic strategy; therefore, there is a continuous search for new therapeutic modalities, for application either alone or in combination. In this regard, gene-based therapy is a new approach that offers hope of improved control of tumors. Intensive research to apply gene therapy for cancer treatment has led to identification of the most important technical and theoretical barriers that need to be overcome for clinical success. One of the central unresolved challenges remains the issue of specific and efficient delivery of genes to target cells or tissues, emphasizing the importance of the gene carrier. Along with different viral and non-viral vector systems, mammalian cells have also been considered as vehicles for delivery of anti-cancer therapeutics. The cell-based delivery approach was introduced as the first attempt to apply gene therapy to cancer treatment, and in general, has followed most of the ups and downs of gene therapy applications, progressing alongside new knowledge gained in this field. As a result, significant progress has been made in some aspects of the cell-based approach, while the development of other essential issues is only just gaining speed. It appears that the initial phase of development of cell-based protocols - the achievement of efficient ex vivo cell loading with therapeutics - has largely been fulfilled. However, the desired efficacy of cell-based strategies in general has not yet been reached, and specificity of tumor homing needs to be improved considerably. There is hope that advances in related scientific fields will promote the utilization of cells as powerful and versatile vehicles for cancer gene therapy.


Does normalized signal intensity of cervical discs on T2 weighted MRI images change in whiplash patients?
We tested the hypothesis that whiplash trauma leads to changes of the signal intensity of cervical discs in T2-weighted images. 50 whiplash patients (18-65 years) were examined within 48h after motor vehicle accident, and again after 3 and 6 months and compared to 50 age- and sex-matched controls. Signal intensity in ROI's of the discs at the levels C2/3 to C7/T1 and the adjacent vertebral bodies were measured on sagittal T2 weighted MR images and normalized using the average of ROI's in fat tissue. The contrast between discs and both adjacent vertebrae was calculated and disc degeneration was graded by the Pfirrmann-grading system. Whiplash trauma did not have a significant effect on the normalized signals from discs and vertebrae, on the contrast between discs and adjacent vertebrae, or on the Pfirrmann grading. However, the contrast between discs and adjacent vertebrae and the Pfirrmann grading showed a strong correlation. In healthy volunteers, the contrast between discs and adjacent vertebrae and Pfirrmann grading increased with age and was dependent on the disc level. We could not find any trauma related changes of cervical disc signal intensities. Normalized signals of discs and Pfirrmann grading changed with age and varied between disc levels with the used MR sequence.


Astaxanthin protects ARPE-19 cells from oxidative stress via upregulation of Nrf2-regulated phase II enzymes through activation of PI3K/Akt.
Oxidative stress on retinal pigment epithelial (RPE) cells is thought to play a crucial role in the development and progression of age-related macular degeneration. Astaxanthin (AST) is a carotenoid that shows significant antioxidant properties. This study was designed to investigate the protective effect of AST on ARPE-19 cells against oxidative stress and the possible underlying mechanism. ARPE-19 cells exposed to different doses of H2O2 were incubated with various concentrations of AST and cell viability subsequently detected with the (4-[3-[4-iodophenyl]-2-4(4-nitrophenyl)-2H-5- tetrazolio-1,3-benzene disulfonate]; WST-1) assay. The apoptosis rate and intracellular levels of reactive oxygen species (ROS) were measured with flow cytometry. NAD(P)H quinine oxidoreductase 1 (NQO1), hemeoxygenase-1 (HO-1), glutamate-cysteine ligase modifier subunit (GCLM), and glutamate-cysteine ligase catalytic subunit (GCLC) expression were examined with real-time PCR and western blotting. The nuclear localization of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) protein and the expression levels of cleaved caspase-3 and protein kinase B proteins were evaluated with western blotting. AST clearly reduced H2O2-induced cell viability loss, cell apoptosis, and intracellular generation of ROS. Furthermore, treatment with AST activated the Nrf2-ARE pathway by inducing Nrf2 nuclear localization. Consequently, Phase II enzymes NQO1, HO-1, GCLM, and GCLC mRNA and proteins were increased. AST inhibited expression of H2O2-induced cleaved caspase-3 protein. Activation of the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) pathway was involved in the protective effect of AST on the ARPE-19 cells. AST protected ARPE-19 cells against H2O2-induced oxidative stress via Nrf2-mediated upregulation of the expression of Phase II enzymes involving the PI3K/Akt pathway.


Linking wound response and inflammation to regeneration in the zebrafish larval fin.
The study of regenerative biology aims to elucidate the innate ability of organisms to replace tissues or organs after they have been removed or damaged. The zebrafish is a powerful model for the analysis of intracellular signalling and cell behaviour and as such has made major contributions to our understanding of regenerative biology. The larval fin fold is an emerging model to understand how different signalling pathways interact to coordinate regeneration. Tissue damage causes the immediate release of signals that initiate wound closure and inflammation. Following this, regenerative cells proliferate and migrate to the damaged area. Each of these processes has been analysed using the larval fin fold model to provide a framework for how fin regeneration takes place. This review gives an overview of the current state of this field with particular emphasis on the different signalling networks that are required during fin fold regeneration.


Comparative interaction networks: bridging genotype to phenotype.
Over the past decade, biomedical research has witnessed an exponential increase in the throughput of the characterization of biological systems. Here we review the recent progress in large-scale methods to determine protein-protein, genetic and chemical-genetic interaction networks. We discuss some of the limitations and advantages of the different methods and give examples of how these networks are being used to study the evolutionary process. Comparative studies have revealed that different types of protein-protein interactions diverge at different rates with high conservation of co-complex membership but rapid divergence of more promiscuous interactions like those that mediate post-translational modifications. These evolutionary trends have consistent genetic consequences with highly conserved epistatic interactions within complex subunits but faster divergence of epistatic interactions across complexes or pathways. Finally, we discuss how these evolutionary observations are being used to interpret cross-species chemical-genetic studies and how they might shape therapeutic strategies. Together, these interaction networks offer us an unprecedented level of detail into how genotypes are translated to phenotypes, and we envision that they will be increasingly useful in the interpretation of genetic and phenotypic variation occurring within populations as well as the rational design of combinatorial therapeutics.


Tobacco use and cessation among veterans recovering from stroke or TIA: a qualitative assessment and implications for rehabilitation.
To understand factors associated with tobacco use and related tobacco cessation among veterans recovering from stroke/transient ischemic attack (TIA) that will facilitate design of a tailored intervention for rehabilitation services. Four focus groups were conducted with veterans who were smokers prior to an incident stroke or TIA along with their spouse or caregiver. Focus groups addressed tobacco use, cessation, and barriers to quitting during the recovery and maintenance periods. Focus group discussions were audiotaped, transcribed, and analyzed using an inductive qualitative method. Twenty-eight veterans and spouses/caregivers participated. Five themes emerged from analysis: existing helpful resources for cessation, existing unhelpful resources, barriers and facilitators to cessation, desired resources for quitting, and association of stroke/TIA with tobacco use. Pharmacotherapy and support from medical providers were perceived as helpful whereas group programs and flyers were perceived as unhelpful. Barriers to quitting included boredom and lack of social support; facilitators included social support and the cost of tobacco products. Vocational and rehabilitation programs were highly desirable resources for quitting. Participants did not perceive their stroke/TIA to be associated with tobacco use. Results identified several issues concerning tobacco use and cessation relevant to patients recovering from stroke/TIA. These results can inform the development of a tailored cessation intervention for integration into rehabilitation and recovery treatment plans for patients with stroke/TIA.


A study of the effect of alpha-linolenic acid on the desaturation of dihomo-gamma-linolenic acid using rat liver homogenates.
The effects of alpha-linolenic acid on delta 5 desaturation of 20: 3 n-6 dihomo-gamma-linolenic acid) to 20: 4 n-6 (arachidonic acid) and on delta 6 desaturation of 18: 2 n-6 (linoleic acid) to 18:3 n-6 (gamma-linolenic acid) were investigated in two different studies using rat liver homogenates. In the first study, rats were fed a semi-synthetic fat-free diet. When unlabelled 18:3 n-3 was added to the incubation of 14C 18:2 or 14C 20:3, the desaturation rates decreased. delta 6 Desaturation was more strongly inhibited than delta 5 desaturation. The position isomer, gamma-linolenic acid, had no significant effect on delta 6 and delta 5 desaturation. In the second study, the rats were fed a diet including alpha-linolenic acid in the form of rapeseed oil containing no erucic acid. The control animals were fed a peanut oil diet. When alpha-linolenic-fed rats were compared to control animals, we found that delta 6 desaturase activity was lower in the alpha-linolenic-fed animals. In contrast, there was no significant effect on the desaturation rate when 14C 20:3 n-6 was used as a substrate. Our results lead to the conclusion that alpha-linolenic acid, when given to rats as dietary rapeseed oil, partly inhibited delta 6 desaturation in vitro, but did not affect significantly subsequent delta 5 desaturation in the biosynthesis of arachidonic acid. The inhibition of the delta 5 desaturation step, when alpha-linolenic acid was added to the incubation, might be due to an unknown competitive mechanism. These observations have been discussed in relation with the current literature.


Persistent allergic rhinitis in older patients: therapies worth trying.
Allergic rhinitis (hayfever) is a common problem found in the geriatric population. This problem should be aggressively treated with environmental control, prescription nasal sprays and/or antihistamines (if tolerated). Allergy immunotherapy can be a very effective treatment for the elderly, without the side effects commonly found when hayfever medication is used.


Involvement of mitochondrial and B-RAF/ERK signaling pathways in berberine-induced apoptosis in human melanoma cells.
The natural isoquinoline alkaloid berberine exhibits a wide spectrum of biological activities including antitumor activity, but its mechanism of action remains to be fully elucidated. Here, we report that berberine induced apoptosis in human melanoma cells, through a process that involved mitochondria and caspase activation. Berberine-induced activation of a number of caspases, including caspases 3, 4, 7, 8, and 9. Pan-caspase inhibitor, z-VAD-fmk, and caspase-8 and caspase-9 inhibitors prevented apoptosis. Berberine also led to the generation of the p20 cleavage fragment of BAP31, involved in directing proapoptotic signals between the endoplasmic reticulum and the mitochondria. Treatment of SK-MEL-2 melanoma cells with berberine induced disruption of the mitochondrial transmembrane potential, release of cytochrome c and apoptosis-inducing factor from the mitochondria to the cytosol, generation of reactive oxygen species (ROS), and a decreased ATP/ADP ratio. Overexpression of bcl-xL by gene transfer prevented berberine-induced cell death, mitochondrial transmembrane potential loss, and cytochrome c and apoptosis-inducing factor release, but not ROS generation. N-acetyl-L-cysteine inhibited the production of ROS, but did not abrogate the berberine-induced apoptosis. Inhibition of extracellular signal-regulated kinase (ERK) phosphorylation, by using the mitogen-activated protein kinase/ERK kinase inhibitor PD98059, and reduction of B-RAF levels by silencing RNA induced cell death of SK-MEL-2 cells, and diminished the berberine concentration required to promote apoptosis. These data show that berberine-induced apoptosis in melanoma cells involves mitochondria and caspase activation, but ROS generation was not essential. Our results indicate that inhibition of B-RAF/ERK survival signaling facilitates the cell death response triggered by berberine.


Abuse disclosures of youth with problem sexualized behaviors and trauma symptomology.
The majority of youth with problem sexualized behaviors (PSB) have substantiated experiences of abuse or exposures to violence (Silovsky & Niec, 2002). Little is known about specific abuse experiences that may differentiate youth with PSB from those without. Few studies have examined the types of abuse associated with post-traumatic stress symptomology. The current study explored two research questions: (1) Do children with PSB differ from children without PSB in terms of their abuse disclosures?; and (2) Are the types of abuse disclosed associated with the child's scores on a post-traumatic stress measure?. Data were analyzed for youth (N = 950) ages 3-18 years who completed a clinical assessment at a child advocacy center in the Midwest during the 2015 calendar year. Youth completed assessments that included a forensic interview and either the Trauma Symptom Checklist for Young Children (TSCYC) for children ages 3-10 years, or the Trauma Symptom Checklist for Children (TSCC) for children ages 11-16 years. Bivariate logistic regression was used to answer the research questions. Findings indicated that youths who disclosed offender to victim fondling were less likely to disclose PSB (OR = 0.460, p = .026), and children exposed to pornography were more likely to disclose PSB (OR = 3.252, p = .001). Additionally, youth who disclosed physical abuse (OR = 1.678, p = .001) or victim to offender sexual contact (OR = 2.242, p = .003) had higher odds of clinically significant trauma scores. Implications for practitioners and future research directions are discussed.


The efficacy and safety of auricular point pressing therapy for knee osteoarthritis: A protocol for systematic review and meta-analysis.
Knee osteoarthritis (KOA) is one of the leading causes of disability. The effectiveness of auricular point pressing therapy for treating KOA remains controversial. This protocol describes the method of a systematic review and meta-analysis evaluating the effectiveness and safety of auricular point pressing therapy for treating KOA. Four English databases (PubMed, Embase, Cochrane Library databases and Web of Science) and 4 Chinese databases (China National Knowledge Infrastructure, Chinese Biomedical Literature Database, VIP Database for Chinese Technical Periodicals, and Wanfang) will be searched. All randomized controlled trials related to auricular point pressing therapy for KOA will be included. Extracted data will include publication details, basic information, demographic data, intervention details and patient outcomes. The primary outcome will be Western Ontario and McMaster Universities Osteoarthritis Index and visual analogue scale. Risk of bias will be assessed using the Cochrane Collaboration tool for assessing risk of bias. Article selection, data extraction and risk of bias assessment will be performed by 2 independent reviewers. If the meta-analysis is precluded, we will conduct a descriptive synthesis using a best-evidence synthesis approach. The strength of recommendations and quality of evidence will be assessed using the Grading of Recommendations Assessment Development. The systematic review will provide evidence to assess the efficacy and safety of auricular point pressing therapy on KOA. The systematic review will provide evidence to assess the efficacy and safety of auricular point pressing therapy for KOA patients. For this review, ethical approval is not required. Patients will not be involved. The findings will be published in a peer-reviewed journal. INPLASY 202220077.


Craniocervical instability in the setting of os odontoideum: assessment of cause, presentation, and surgical outcomes in a series of 279 cases.
Our clinical understanding of os odontoideum (OO) remains incomplete. Congenital and traumatic causes have been proposed and advocated. Clinical presentations range from asymptomatic to axial pain to myelopathy or vertebral-basilar ischemia. A consensus for surgical management exists for those found to have an unstable atlantoaxial complex or symptomatic cranial-vertebral junction compression. To evaluate the clinical presentation and surgical outcomes of patients with OO and an unstable atlantoaxial complex or symptomatic cranial-vertebral junction compression. Patients with a diagnosis of OO who underwent surgical management were included. Patients were excluded on the basis of previous C2 fracture, Fielding diagnostic criteria, and inadequate follow-up. History of trauma and presenting symptoms were assessed. Clinical and neurological improvements were measured with the use of patient satisfaction scores and the Japanese Orthopaedic Association scores. Fusion status was documented with the use of radiographs and computed tomographic imaging. Of 279 patients, 112 reported a history of cranial-vertebral junction trauma, whereas 28 were diagnosed with congenital malformations. Clinically, 84.9% of patients presented with myelopathy, with pain presented in 42.6%. Atlantoaxial fixation was performed in 240 patients, occiput-to-C2 fixation in 35 patients, and extended occipito-cervical fixation in 4 patients. Mean follow-up was 40.3 months. Complications were reported in 2.4% of patients. Japanese Orthopaedic Association scores improved from a preoperative mean of 12.4 to 14.8. Two hundred thirty-five patients (77.7%) improved, with 30 patients experiencing no change in symptoms and 14 patients deteriorating. Fusion was achieved in 96.8% of patients. Our data reveal that surgical treatment for OO using the indications and techniques delineated is associated with high satisfaction rates, improved functional scores, and high fusion rates with low complication rates.


Brewer's yeast efficiently degrades phytate phosphorus in a corn-soybean meal diet during soaking treatment.
Microbes such as yeast and Aspergillus are known to produce phytase, and Aspergillus phytase has been used as a feed additive for improving phytate-phosphorus bioavailability in monogastric animals. We measured phytase activity in some by-products from fermented food and beverage productions by yeast and Aspergillus. The phytase activity was as high as 3577 and 2225 PU/kg DM in raw and dried brewer's yeasts, respectively. On the other hand, the phytase activity was approximately 400 PU/kg DM in white-wine yeast and red-wine yeast. The phytase activity was further low in natto (fermented soybean) residue, soy sauce cake, rice brewer's grain and the activity was not detected in dried corn-barley distiller's grain with soluble and sweet-potato distiller's residue. The stability of phytase against pepsin was much lower in the brewer's yeast than in an Aspergillus phytase preparation. On the other hand, the addition of raw brewer's yeast effectively degraded phytate phosphorus in a corn-soybean meal diet during soaking. These results suggest that phytase in the examined by-products is not suitable for the phytase source of conventional diets, but that the soaking treatment with a raw brewer's yeast is an alternative method for improving phytate-phosphorus bioavailability in corn-soybean meal diets for pigs.


Optimization of size and encapsulation efficiency of 5-FU loaded chitosan nanoparticles by response surface methodology.
The objective of this study was to optimize the particle size and encapsulation efficiency of chitosan nanoparticles loaded with 5-Fluorouracil (5-FU) by response surface methodology. Nanoparticles were prepared by ionic gelation method from chitosan and penta sodium triphosphate (TPP) at different combinations of chitosan viscosity, chitosan concentration and chitosan/TPP mass ratio according to the Box-Behnken experimental design. The particle size and encapsulation efficiency of prepared particles were measured by dynamic light scattering and UV spectroscopy, respectively, and the obtained data were subjected to multiple linear regression analysis followed by multi-attribute utility analysis to obtain a model for prediction of the optimum response. The optimum conditions for the production of 5-FU loaded chitosan nanoparticles were found to be low viscosity chitosan 0.5-1 mg/mL, middle viscosity chitosan 0.5-0.8 mg/mL and high viscosity chitosan 0.5-0.75 mg/mL and chitosan/TPP mass ratio of 4 or 6, yielding nanoparticles at the average diameter range of 114-188 nm and encapsulation efficiencies between 42-55%.


Gating by inhibition during top-down control of willed attention.
In natural settings, the control of attention may be influenced both by external information as well as internal decision-making processes driven by intent (e.g. free will). In past studies of spatial attention, we and others have developed experimental paradigms that permit individuals to choose where to direct their attention on a trial-by-trial basis in the absence of instructive external cues - we term this willed attention. Here we investigate the electrophysiological correlates of willed attention by recording EEG activity when subjects decided to focus covert attention on one of two lateralized target locations versus when they decided to maintain attention at fixation. Independent of the direction of attention, decisions to attend, relative to decisions not to attend, resulted in significant increases in both frontal theta (4-7 Hz) power and central alpha (8-13 Hz) power. We found that focusing spatial attention, as indexed by occipital alpha lateralization was predicted across subjects by the decision-related alpha increases over central scalp regions, but not changes in frontal theta power. This finding is interpreted in terms of the Gating by Inhibition model, where the central alpha EEG signals reflect cortical inhibition of decision processes that lead to the expression of willed attention.


In vivo space radiation-induced non-targeted responses: late effects on molecular signaling in mitochondria.
The lack of clear knowledge about space radiation-induced biological effects has been singled out as the most important factor limiting the prediction of radiation risk associated with human space exploration. The expression of space radiation-induced non-targeted effects is thought to impact our understanding of the health risks associated with exposure to low fluences of particulate radiation encountered by astronauts during prolonged space travel. Following a brief review of radiation-induced bystander effects and the growing literature for the involvement of oxidative metabolism in their expression, we show novel data on the induction of in vivo non-targeted effects following exposure to 1100 MeV/nucleon titanium ions. Analyses of proteins by two-dimensional gel electrophoresis in non-targeted liver of cranially-irradiated Sprague Dawley rats revealed that the levels of key proteins involved in mitochondrial fatty acid metabolism are decreased. In contrast, those of proteins involved in various cellular defense mechanisms, including antioxidation, were increased. These data contribute to our understanding of the mechanisms underlying the biological responses to space radiation, and support the involvement of mitochondrial processes in the expression of radiation induced non-targeted effects. Significantly, they reveal the cross-talk between propagated stressful effects and induced adaptive responses. Together, with the accumulating data in the field, our results may help reduce the uncertainty in the assessment of the health risks to astronauts. They further demonstrate that 'network analyses' is an effective tool towards characterizing the signaling pathways that mediate the long-term biological effects of space radiation.


A Low-Power Gateable Vernier Ring Oscillator Time-to-Digital Converter for Biomedical Imaging Applications.
In this paper, a high resolution, high precision and ultra-low power consumption time-to-digital converter (TDC) is presented. The proposed TDC is based on the gateable Vernier ring oscillator architecture. Fine resolution is achieved through two ring oscillators arranged in the Vernier configuration. This TDC employs a single-transition end-of-conversion detection circuit and turns off the ring oscillators whenever the conversion is completed to reduce power consumption. The prototype chip is fabricated in a standard 130 nm digital CMOS process and its area is only 0.03 mm(2). Using a 1.2 V supply, the TDC achieves a resolution of 7.3 ps, a single-shot precision of 1.0LSB, and an average power consumption of 1.2 mW. A root-mean-square integral nonlinearity (INL) of 1.2 LSB is obtained with the help of an INL look-up-table calibration. Compared to previously reported ring-oscillator based TDCs, the proposed design achieves the lowest power consumption to date.


Rearrangements of 2.5 kilobases of noncoding DNA from the Drosophila even-skipped locus define predictive rules of genomic cis-regulatory logic.
Rearrangements of about 2.5 kilobases of regulatory DNA located 5' of the transcription start site of the Drosophila even-skipped locus generate large-scale changes in the expression of even-skipped stripes 2, 3, and 7. The most radical effects are generated by juxtaposing the minimal stripe enhancers MSE2 and MSE3 for stripes 2 and 3 with and without small "spacer" segments less than 360 bp in length. We placed these fusion constructs in a targeted transformation site and obtained quantitative expression data for these transformants together with their controlling transcription factors at cellular resolution. These data demonstrated that the rearrangements can alter expression levels in stripe 2 and the 2-3 interstripe by a factor of more than 10. We reasoned that this behavior would place tight constraints on possible rules of genomic cis-regulatory logic. To find these constraints, we confronted our new expression data together with previously obtained data on other constructs with a computational model. The model contained representations of thermodynamic protein-DNA interactions including steric interference and cooperative binding, short-range repression, direct repression, activation, and coactivation. The model was highly constrained by the training data, which it described within the limits of experimental error. The model, so constrained, was able to correctly predict expression patterns driven by enhancers for other Drosophila genes; even-skipped enhancers not included in the training set; stripe 2, 3, and 7 enhancers from various Drosophilid and Sepsid species; and long segments of even-skipped regulatory DNA that contain multiple enhancers. The model further demonstrated that elevated expression driven by a fusion of MSE2 and MSE3 was a consequence of the recruitment of a portion of MSE3 to become a functional component of MSE2, demonstrating that cis-regulatory "elements" are not elementary objects.


A repeated measures experiment of green exercise to improve self-esteem in UK school children.
Exercising in natural, green environments creates greater improvements in adult's self-esteem than exercise undertaken in urban or indoor settings. No comparable data are available for children. The aim of this study was to determine whether so called 'green exercise' affected changes in self-esteem; enjoyment and perceived exertion in children differently to urban exercise. We assessed cardiorespiratory fitness (20 m shuttle-run) and self-reported physical activity (PAQ-A) in 11 and 12 year olds (n = 75). Each pupil completed two 1.5 mile timed runs, one in an urban and another in a rural environment. Trials were completed one week apart during scheduled physical education lessons allocated using a repeated measures design. Self-esteem was measured before and after each trial, ratings of perceived exertion (RPE) and enjoyment were assessed after completing each trial. We found a significant main effect (F (1,74), = 12.2, p<0.001), for the increase in self-esteem following exercise but there was no condition by exercise interaction (F (1,74), = 0.13, p = 0.72). There were no significant differences in perceived exertion or enjoyment between conditions. There was a negative correlation (r = -0.26, p = 0.04) between habitual physical activity and RPE during the control condition, which was not evident in the green exercise condition (r = -0.07, p = 0.55). Contrary to previous studies in adults, green exercise did not produce significantly greater increases in self-esteem than the urban exercise condition. Green exercise was enjoyed more equally by children with differing levels of habitual physical activity and has the potential to engage less active children in exercise.


Cognitive sequelae of endocrine therapy in women treated for breast cancer: a meta-analysis.
Evidence suggests anti-estrogen endocrine therapy (ET) is associated with adverse cognitive effects; however, findings are based on small samples and vary in the cognitive abilities affected. We conducted a meta-analysis to quantitatively synthesize the evidence. Electronic databases were searched in November 2016. Fourteen studies totaling 911 BC patients on aromatase inhibitors (AIs) or tamoxifen (TAM) and 911 controls (i.e., non-cancer controls and BC controls not using ET) were included. Neuropsychological tests were categorized into six domains. Effect sizes were computed to compare (1) ET patients versus controls and (2) TAM patients versus AI patients. In cross-sectional comparisons, ET patients performed worse than control groups on verbal learning/memory, visual learning/memory, frontal executive function, and processing speed, but did not differ on psychomotor efficiency or visuospatial function. Subgroup analyses revealed that verbal learning/memory was the only domain where ET patients performed worse than both non-cancer and BC controls. In other domains, ET patients and BC controls performed equivalently. Regarding change from pre-treatment performance, ET patients did not differ from controls on any domain. TAM and AI patients did not from one another differ overall; however, subgroup analyses indicated that TAM patients performed better than non-steroidal AI patients on several domains, but showed few performance differences relative to steroidal AI patients. Verbal learning/memory was the only domain where ET patients performed worse than both non-cancer and BC controls, suggesting specific adverse effects on this domain. Additional studies assessing change from pre-treatment performance and differences between steroidal and non-steroidal AIs are warranted.


Solid state and solution conformation of Boc-L-Met-Aib-L-Phe-OMe. Beta-turn conformation of a sequence related to an active chemotactic peptide analog.
The conformation of the peptide Boc-L-Met-Aib-L-Phe-OMe has been studied in the solid state and solution by X-ray diffraction and 1H n.m.r., respectively. The peptide differs only in the N-terminal protecting group from the biologically active chemotactic peptide analog formyl-L-Met-Aib-L-Phe-OMe. The molecules adopt a type-II beta-turn in the solid state with Met and Aib as the corner residues (phi Met = -51.8 degrees, psi Met = 139.5 degrees, phi Aib = 58.1 degrees, psi Aib = 37.0 degrees). A single, weak 4----1 intramolecular hydrogen bond is observed between the Boc CO and Phe NH groups (N---O 3.25 A, N-H---O 128.4 degrees). 1H n.m.r. studies, using solvent and temperature dependencies of NH chemical shifts and paramagnetic radical induced line broadening of NH resonances, suggest that the Phe NH is solvent shielded in CDCl3 and (CD3)2SO. Nuclear Overhauser effects observed between Met C alpha H and Aib NH protons provide evidence of the occurrence of Met-Aib type-II beta-turns in these solvents.


The molecular ecologist's guide to expressed sequence tags.
Genomics and bioinformatics have great potential to help address numerous topics in ecology and evolution. Expressed sequence tags (ESTs) can bridge genomics and molecular ecology because they can provide a means of accessing the gene space of almost any organism. We review how ESTs have been used in molecular ecology research in the last several years by providing sequence data for the design of molecular markers, genome-wide studies of gene expression and selection, the identification of candidate genes underlying adaptation, and the basis for studies of gene family and genome evolution. Given the tremendous recent advances in inexpensive sequencing technologies, we predict that molecular ecologists will increasingly be developing and using EST collections in the years to come. With this in mind, we close our review by discussing aspects of EST resource development of particular relevance for molecular ecologists.


Drug-binding properties of rat alpha-foetoprotein. Specificities of the phenylbutazone-binding and warfarin-binding sites.
Rat alpha-foetoprotein (alpha-FP) strongly binds the drugs warfarin and phenylbutazone, as does albumin; however, the binding sites for the two drugs seemed to be different. This possibility and the specificity of this/these drug-binding site(s) of rat alpha-FP were investigated by competitive protein-binding experiments with a variety of drugs, representing different pharmacological groups, and biomolecules that are strongly bound by the foetal protein and that are suspected to play a specific role during foetal development. The binding mechanisms were further investigated by using comparisons between computer-derived theoretical displacement curves and experimental points in order to distinguish different possible binding models. The results indicate: that warfarin and phenylbutazone are bound at two distinct sites on rat alpha-FP and that a negative modulatory effect is exerted between the two sites; that the degree of specificity of these two drug-binding sites is different, since the warfarin-binding site appears to be specific for the binding of coumarinic and anthranilic drugs whereas that for phenylbutazone is able to bind substances of very varied chemical structure and is more hydrophobic; that the phenylbutazone-binding site is the site that binds oestrogens that thyroid hormones and, probably, fatty acids and bilirubin are bound at (an)other site(s) but exert negative modulatory effects on phenylbutazone binding. The nature of the different binding areas of rat alpha-FP is compared with that of those already proposed for albumin. The potential risks of toxicity of such interactions between drugs and/or biomolecules on foetal development are also discussed.


Imaging of intracellular metal partitioning in marine diatoms exposed to metal pollution: consequences to cellular toxicity and metal fate in the environment.
This study investigates the metal content and compartmentalization changes in whole cells of diatom Coscinodiscus eccentricus exposed to metal overload, examining consequences to cellular toxicity, tolerance mechanisms, and metal fate in the environment. Cells exposed to Ni, Cu and Zn were analysed using nuclear microprobe techniques. Particle induced X-ray emission (PIXE), Rutherford backscattering spectrometry (RBS), and scanning transmission ion microscopy (STIM) were used simultaneously to obtain high-resolution imaging of morphological and quantitative elemental distribution data. Elemental partitioning within cell compartments, such as cell wall, cytoplasm and major organelles, was assessed. Diatoms clearly responded to excess metal levels, by changing cytoplasm morphology, concentrating added metals, and altering Fe transport mechanisms. Different metal accumulation patterns indicated high susceptibility to Cu, retained in the cytoplasm, and detoxification capability for Ni and Zn, mobilized to the vacuole. Iron and Zn were accumulated in the siliceous wall. Different metal distributions within the cell imply distinct environmental fates, Cu and Ni remain available with potential for biomagnification through the food web, whereas Fe and Zn are deposited at the bottom through frustule sedimentation.


A quasi-glottogram signal.
A novel, noninvasive experiment is proposed that reliably shows the strength of glottal oscillations. The quasi-glottogram (QGG) signal is generated from a microphone array that is trained to approximate the electroglottogram signal. The QGG may be useful to improve estimates of whether speech is voiced, to quantify partial voicing, and to reduce the phoneme effect when measuring the amplitude of speech signals. The technique is well adapted to the generation of text-to-speech systems, as it allows an estimate of the glottal flow during undisturbed, natural speech. For prosody studies, it can be used to provide an estimate of amplitude which is relatively unaffected by changes in phonemes, and is at least as reliable as standard estimators of amplitude.


Traumatic tarsal luxation repaired without internal fixation in three horses and three ponies.
Three horses and three ponies were treated for traumatic luxation of the proximal intertarsal or tarsometatarsal joint. In each case, there were fractures of one or more tarsal bones. Three of the animals had disruption of one of the collateral ligaments. Five animals were treated by closed reduction and external coaptation. One pony was treated by open curettage, a cancellous bone graft, and cast immobilization. No internal fixation was used in any of these animals. From this series of cases, it would appear that closed reduction and external coaption alone is adequate treatment for most cases of tarsal luxation.


Using the Generic Genome Browser (GBrowse).
A genome browser is software that allows users to visualize DNA, protein, or other sequence features within the context of a reference sequence, such as a chromosome or contig. The Generic Genome Browser (GBrowse) is an open-source browser developed as part of the Generic Model Organism Database project (Stein et al., 2002). GBrowse can be configured to display genomic sequence features for any organism and is the browser used for the model organisms Drosophila melanogaster (Grumbling and Strelets, 2006) and Caenorhabditis elegans (Schwarz et al., 2006), among others. The software package can be downloaded from the Web and run on a Windows, Mac OS X, or Unix-type system. Version 1.64, as described in the original protocol, was released in November 2005, but the software is under active development and new versions are released about every six months. This update includes instructions on updating existing data sources with new files from NCBI.


Imaging of Chlorophyll a Fluorescence in Natural Compound-Induced Stress Detection.
Imaging of chlorophyll a fluorescence (CFI) represents an easy, precise, fast and non-invasive technique that can be successfully used for discriminating plant response to phytotoxic stress with reproducible results and without damaging the plants. The spatio-temporal analyses of the fluorescence images can give information about damage evolution, secondary effects and plant defense response. In the last years, some studies about plant natural compounds-induced phytotoxicity have introduced imaging techniques to measure fluorescence, although the analysis of the image as a whole is often missed. In this paper we, therefore, evaluated the advantages of monitoring fluorescence images, presenting the physiological interpretation of different possible combinations of the most relevant parameters linked to fluorescence emission and the images obtained.


Activation of the Campylobacter jejuni FlgSR two-component system is linked to the flagellar export apparatus.
Activation of sigma(54)-dependent gene expression essential for formation of flagella in Campylobacter jejuni requires the components of the inner membrane-localized flagellar export apparatus and the FlgSR two-component regulatory system. In this study, we characterized the FlgS sensor kinase and how activation of the protein is linked to the flagellar export apparatus. We found that FlgS is localized to the C. jejuni cytoplasm and that His141 of FlgS is essential for autophosphorylation, phosphorelay to the cognate FlgR response regulator, motility, and expression of sigma(54)-dependent flagellar genes. Mutants with incomplete flagellar export apparatuses produced wild-type levels of FlgS and FlgR, but they were defective for signaling through the FlgSR system. By using genetic approaches, we found that FlgSR activity is linked to and downstream of the flagellar export apparatus in a regulatory cascade that terminates in expression of sigma(54)-dependent flagellar genes. By analyzing defined flhB and fliI mutants of C. jejuni that form flagellar export apparatuses that are secretion incompetent, we determined that formation of the apparatus is required to contribute to the signal sensed by FlgS to terminate in activation of expression of sigma(54)-dependent flagellar genes. Considering that the flagellar export apparatuses of Escherichia coli and Salmonella species influence sigma(28)-dependent flagellar gene expression, our work expands the signaling activity of the apparatuses to include sigma(54)-dependent pathways of C. jejuni and possibly other motile bacteria. This study indicates that these apparatuses have broader functions beyond flagellar protein secretion, including activation of essential two-component regulatory systems required for expression of sigma(54)-dependent flagellar genes.


Formation of membrane ridges and scallops by the F-BAR protein Nervous Wreck.
Eukaryotic cells are defined by extensive intracellular compartmentalization, which requires dynamic membrane remodeling. FER/Cip4 homology-Bin/amphiphysin/Rvs (F-BAR) domain family proteins form crescent-shaped dimers, which can bend membranes into buds and tubules of defined geometry and lipid composition. However, these proteins exhibit an unexplained wide diversity of membrane-deforming activities in vitro and functions in vivo. We find that the F-BAR domain of the neuronal protein Nervous Wreck (Nwk) has a novel higher-order structure and membrane-deforming activity that distinguishes it from previously described F-BAR proteins. The Nwk F-BAR domain assembles into zigzags, creating ridges and periodic scallops on membranes in vitro. This activity depends on structural determinants at the tips of the F-BAR dimer and on electrostatic interactions of the membrane with the F-BAR concave surface. In cells, Nwk-induced scallops can be extended by cytoskeletal forces to produce protrusions at the plasma membrane. Our results define a new F-BAR membrane-deforming activity and illustrate a molecular mechanism by which positively curved F-BAR domains can produce a variety of membrane curvatures. These findings expand the repertoire of F-BAR domain mediated membrane deformation and suggest that unique modes of higher-order assembly can define how these proteins sculpt the membrane.


[Radical surgical treatment of heart myxoma].
Experience of surgical treatment of 362 patients with the heart myxoma (HM) was summarized. The expediency of application of various approaches in the left auricle (LA) myxoma was analyzed. Significant advantages of application of the approach through the right auricle to reduce the danger of the tumor fragmentation while its excision were noted. There was elaborated and introduced transbiauricle approach to the LA tumor, application of which secures minimal risk of the tumor fragmentation as well as the possibility of adequate heart revision conduction and the processing of full value of tumor bed for any localization. In terms of observation up to 30 years in 5 (17%) of patients the HM recurrency had occurred. Application of the method, which envisages excision of part of interatrial septum, where HM is fixed, in its all thickness with subsequent suturing and plasty of defect using the patch, making additional processing of the tumor bed with iodine solution and diathermocoagulation, had permitted to reduce the recurrency of the disease from 4.3 to 0.5%.


Prominent amphibian (Xenopus laevis) tadpole type III interferon response to the frog virus 3 ranavirus.
Ranaviruses (Iridoviridae) are posing an increasing threat to amphibian populations, with anuran tadpoles being particularly susceptible to these viral infections. Moreover, amphibians are the most basal phylogenetic class of vertebrates known to possess both type I and type III interferon (IFN)-mediated immunity. Moreover, little is known regarding the respective roles of the IFN mediators in amphibian antiviral defenses. Accordingly, we transcriptionally and functionally compared the amphibian Xenopus laevis type I (IFN) and III (IFN-λ) IFNs in the context of infections by the ranavirus frog virus 3 (FV3). X. laevis IFN and IFN-λ displayed distinct tissue expression profiles. In contrast to our previous findings that X. laevis tadpoles exhibit delayed and modest type I IFN responses to FV3 infections compared to the responses of adults, here we report that tadpoles mount timely and robust type III IFN gene responses. Recombinant forms of these cytokines (recombinant X. laevis IFN [rXlIFN] and rXlIFN-λ) elicited antiviral gene expression in the kidney-derived A6 cell line as well as in tadpole leukocytes and tissues. However, rXlIFN-λ was less effective than rXlIFN in preventing FV3 replication in A6 cells and tadpoles and inferior at promoting tadpole survival. Intriguingly, FV3 impaired A6 cell and tadpole kidney type III IFN receptor gene expression. Furthermore, in A6 cultures rXlIFN-λ conferred equal or greater protection than rXlIFN against recombinant viruses deficient for the putative immune evasion genes, the viral caspase activation and recruitment domain (vCARD) or a truncated vIF-2α gene. Thus, in contrast to previous assumptions, tadpoles possess intact antiviral defenses reliant on type III IFNs, which are overcome by FV3 pathogens. Anuran tadpoles, including those of Xenopus laevis, are particularly susceptible to infection by ranavirus such as FV3. We investigated the respective roles of X. laevis type I and type III interferons (IFN and IFN-λ, respectively) during FV3 infections. Notably, tadpoles mounted timely and more robust IFN-λ gene expression responses to FV3 than adults, contrasting with the poorer tadpole type I IFN responses. However, a recombinant X. laevis IFN-λ (rXlIFN-λ) conferred less protection to tadpoles and the A6 cell line than rXlIFN, which may be explained by the FV3 impairment of IFN-λ receptor gene expression. The importance of IFN-λ in tadpole anti-FV3 defenses is underlined by the critical involvement of two putative immune evasion genes in FV3 resistance to IFN- and IFN-λ-mediated responses. These findings challenge the view that tadpoles have defective antiviral immunity and suggest, rather, that their antiviral responses are predominated by IFN-λ responses, which are overcome by FV3.


'Schizoid' personality in childhood and adult life. II: Adult adjustment and the continuity with schizotypal personality disorder.
In a controlled follow-up study into adulthood of 32 children diagnosed 'schizoid', three-quarters fulfilled DSM-III criteria for schizotypal personality disorder and two developed schizophrenia. Overall their psychosocial adjustment was somewhat, but not markedly, worse than that of other attenders at a child psychiatry clinic, although as a group they remained more solitary, lacking in empathy, oversensitive, with odd styles of communicating, and often with circumscribed interests.


Multifunctional targeting daunorubicin plus quinacrine liposomes, modified by wheat germ agglutinin and tamoxifen, for treating brain glioma and glioma stem cells.
Most anticancer drugs are not able to cross the blood-brain barrier (BBB) effectively while surgery and radiation therapy cannot eradicate brain glioma cells and glioma stem cells (GSCs), hence resulting in poor prognosis with high recurrence rates. In the present study, a kind of multifunctional targeting daunorubicin plus quinacrine liposomes was developed for treating brain glioma and GSCs. Evaluations were performed on in-vitro BBB model, murine glioma cells, GSCs, and GSCs bearing mice. Results showed that the multifunctional targeting daunorubicin plus quinacrine liposomes exhibited evident capabilities in crossing the BBB, in killing glioma cells and GSCs and in diminishing brain glioma in mice. Action mechanism studies indicated that the enhanced efficacy of the multifunctional targeting drugs-loaded liposomes could be due to the following aspects: evading the rapid elimination from blood circulation; crossing the BBB effectively; improving drug uptake by glioma cells and GSCs; down-regulating the overexpressed ABC transporters; inducing apoptosis of GSCs via up-regulating apoptotic receptor/ligand (Fas/Fasl), activating apoptotic enzymes (caspases 8, 9 and 3), activating pro-apoptotic proteins (Bax and Bok), activating tumor suppressor protein (P53) and suppressing anti-apoptotic proteins (Bcl-2 and Mcl-1). In conclusion, the multifunctional targeting daunorubicin plus quinacrine liposomes could be used as a potential therapy for treating brain glioma and GSCs.


The first laminin G-type domain in the SHBG-like region of protein S contains residues essential for activation of the receptor tyrosine kinase sky.
Vitamin K-dependent protein S and the product of growth-arrest-specific gene 6 (Gas6) both possess the ability to phosphorylate members of the Axl/Sky subfamily of receptor tyrosine kinases. However, Gas6 appears to be the bona fide ligand for these receptors in man, as human protein S has been demonstrated to activate murine Sky but not the human orthologue. In contrast, bovine protein S is able to stimulate human Sky despite its high degree of sequence identity with human protein S. The domain organisations of protein S and Gas6 are virtually identical and the C-terminal SHBG-like region, containing two globular (G) domains, has been shown to play a crucial role in the receptor stimulation. In order to further localise the area responsible for the interaction, a number of protein chimeras were used to stimulate human Sky. Each chimera had one part of the human protein S SHBG-like region replaced by the corresponding part of bovine protein S or human Gas6. We found that human protein S may indeed activate human Sky but only above physiological plasma concentrations. The human-bovine protein S chimeras provided new information implying that the first G domain contains critical residues for the interaction with the Sky receptor. Moreover, these residues do not seem to be clustered but rather to be distributed at various positions in the first G domain.