How to measure reproductive success?
To date there have been few empirical comparisons between alternative methods for measuring reproductive success (RS). We consider the pros and cons of alternative measures of RS to provide guidance for the design of field studies in human behavioral ecology. We compare cross-sectional measures that count offspring alive at the time of the interview and retrospective measures that require data on offspring age at death or censoring. We consider analyses that include adult women (yielding age-specific estimates of RS) as well as analyses restricted to postreproductive women (yielding data on lifetime RS). These methods are applied to reproductive data for the Dogon of Mali, West Africa.


Metastatic Chordoma: Report of the Two Cases and Review of the Literature.
Chordomas are rare malignant bone tumours with a predilection for the axial skeleton, especially the sacrum and skull base. Median survival in patients with metastatic disease is usually dismal. Treatment is challenging due to the propensity for local recurrence, metastatic disease as well as lack of clear consensus regarding the optimal management. Our case report highlights two cases of sacral chordoma with locally recurrent and widespread metastatic disease, stable on molecular targeted therapy.


Testes exhibit elevated expression of calcitonin gene-related peptide receptor component protein.
Calcitonin gene-related peptide (CGRP) receptor component protein (RCP) is a novel protein that modulates CGRP responsiveness in a variety of cell types. Using probes based on the isolation of CGRP-RCP complementary DNA (cDNA) from a guinea pig organ of Corti cDNA library, we cloned human (h) and mouse (m) CGRP-RCP cDNAs, both of which encode 148-residue proteins that at the amino acid levels are approximately 88% identical to each other and to the 146-residue guinea pig CGRP-RCP. Northern blot analysis confirmed the presence of CGRP-RCP messenger RNA in all of the human and mouse tissues tested. In these human tissues, hCGRP-RCP messenger RNA (major band at approximately 3.1 kb, minor band at approximately 7.5 kb) was most prevalent in the testis. In the mouse, the highest abundance of CGRP-RCP RNA was clearly in the testis (major band at approximately 1.6 kb, minor band at approximately 1.1 kb). Based on this tissue distribution of RNA, we sought to identify the cells in the murine testis that contained CGRP-RCP protein. Numerous antisera generated against hCGRP-RCP, including one to recombinant hCGRP-RCP, exhibited strong immunoreactivity localized to the head region of spermatozoa. No CGRP-RCP immunoreactivity was observed in other cells at less mature stages of sperm maturation, in Sertoli or interstitial (Leydig) cells, or in human spermatozoa. Murine epididymal (mature) spermatozoa exhibited CGRP-RCP immunoreactivity identical to that of testicular spermatozoa. Spermatozoa that underwent an experimentally induced acrosome reaction (acrosomal discharge) lost their CGRP-RCP immunoreactivity. Therefore, it appears that CGRP-RCP is associated with the acrosome, suggesting that it may play an important role in reproduction.


Cyclic patterns of cerebral malaria admissions in Papua New Guinea for the years 1987-1996.
Data on the dynamics of malaria incidence, admissions and mortality and their best possible description are very important to better forecast and assess the implementation of programmes to register, monitor (e.g. by remote sensing) and control the disease, especially in endemic zones. Semi-annual and seasonal cycles in malaria rates have been observed in various countries and close similarity with cycles in the natural environment (temperature, heliogeophysical activity, etc.), host immunity and/or virulence of the parasite suggested. This study aimed at confirming previous results on malaria cyclicity by exploring whether trans-year and/or multiannual cycles might exist. The exploration of underlying chronomes (time structures) was done with raw data (without smoothing) by linear and nonlinear parametric regression models, autocorrelation, spectral (Fourier) and periodogram regression analysis. The strongest cyclical patterns of detrended malaria admissions were (i) annual period of 1·0 year (12 months or seasonality); (ii) quasi-biennial cycle of about 2·25 years; and (iii) infrannual, circadecennial cycle of about 10·3 years. The seasonal maximum occurred in May with the minimum in September. Notably, these cycles corresponded to similar cyclic components of heliogeophysical activity such as sunspot seasonality and solar activity cyclicities and well-known climate/weather oscillations. Further analyses are thus warranted to investigate such similarities. In conclusion, multicomponent cyclical dynamics of cerebral malaria admissions in Papua New Guinea were observed thus allowing more specific analyses and modelling as well as correlations with environmental factors of similar cyclicity to be explored. Such further results might also contribute to and provide more precise estimates for the forecasting and prevention, as well as the better understanding, of the dynamics and aetiology of this vector-borne disease.


African countries propose a regional oral health strategy: The Dakar Report from 1998.
It is clear that the African region faces a number of serious oral diseases, either because of their high prevalence or because of the severe tissue damage or death that can occur. Previous approaches to oral health in Africa have failed to recognise the epidemiological priorities of the region or to identify reliable and appropriate strategies to assess them. Efforts have consisted of an unplanned, ad hoc and spasmodic evolution of curative oral health services. This document focuses on the most severe oral problems that people have to live with like noma, oral cancer and the oral consequences of HIV/AIDS infection. It proposes a strategy for assisting member states and partners to identify priorities and interventions at various levels of the health system, particularly at the district level. The strategy aims at strengthening the capacity of countries to improve community oral health by effectively using proven interventions to address specific oral health needs. The strategy identifies five main 'programmatic areas', including (i) the development of national oral health strategies and implementation plans, (ii) integration of oral health in other programmes, (iii) delivery of effective and safe oral health services, (iv) regional approach to education and training for oral health, and (v) development of effective oral health management information systems. Many of the programmatic areas share similar characteristics described as a 'strategic orientation'. These strategic orientations give effect to the concepts of advocacy, equity, quality, partnership, operational research, communication and capacity building. The WHO Regional Committee for Africa (RC) is invited to review the proposed oral health strategy for the African region for the period 1999-2008 and provide an orientation for the improvement of oral health in member states in the region.


Prophage induction and expression of prophage-encoded virulence factors in group A Streptococcus serotype M3 strain MGAS315.
The genome of the highly virulent group A Streptococcus (GAS) serotype M3 strain MGAS315 has six prophages that encode six proven or putative virulence factors. We examined prophage induction and expression of prophage-encoded virulence factors by this strain under in vitro conditions inferred to approximate in vivo conditions. Coculture of strain MGAS315 with Detroit 562 (D562) human epithelial pharyngeal cells induced the prophage encoding streptococcal pyrogenic exotoxin K (SpeK) and extracellular phospholipase A(2) (Sla) and the prophage encoding streptodornase (Sdn). Increased gene copy numbers after induction correlated with increased speK, sla, and sdn transcript levels. Although speK and sla are located contiguously in prophage Phi315.4, these genes were transcribed independently. Whereas production of immunoreactive SpeK was either absent or minimal during coculture of GAS with D562 cells, production of immunoreactive Sla increased substantially. In contrast, despite a lack of induction of the prophage encoding speA during coculture of GAS with D562 cells, the speA transcript level and production of immunoreactive streptococcal pyrogenic exotoxin A (SpeA) increased. Exposure of strain MGAS315 to hydrogen peroxide, an oxidative stressor, induced the prophage encoding mitogenic factor 4 (MF4), and there was a concomitant increase in the mf4 transcript. All prophages of strain MGAS315 that encode virulence factors were induced during culture with mitomycin C, a DNA-damaging agent. However, the virulence factor gene transcript levels and production of the encoded proteins decreased after mitomycin C treatment. Taken together, the results indicate that a complex relationship exists among environmental culture conditions, prophage induction, and production of prophage-encoded virulence factors.


TLR2 agonists enhance CD8+Foxp3+ regulatory T cells and suppress Th2 immune responses during allergen immunotherapy.
Pam3CSK4, a synthetic TLR2 ligand, has been shown to expand CD4+ regulatory T cells (Treg cells). Less is known about the function of CD8+ Treg cells than about the function of CD4+ Treg cells generated during allergen-specific immunotherapy (IT). This study investigated whether Dermatophagoides pteronyssinus-specific IT could expand the CD8+CD25+Foxp3+ Treg population and whether Pam3CSK4 could enhance the Treg population. PBMCs were isolated from healthy control subjects and from mite-sensitive asthmatic patients during IT at three specific times: before IT and 6 mo and 1 y after the maximum-tolerated dose. This study was performed without a placebo-controlled group. D. pteronyssinus-specific IT induced a significant increase in CD8+Foxp3+ Treg cells expressing intracellular IL-10 and granzyme B. Costimulation of PBMCs with Pam3CSK4 and D. pteronyssinus 2 expanded the CD8+CD25+Foxp3+ Treg population and inhibited D. pteronyssinus 2-induced IL-4 production. Pam3CSK4-treated CD8+CD25+ Treg cells directly suppressed CD4+ T cell proliferation by cell-contact inhibition. TUNEL revealed that CD8+CD25+ Treg cells, but not CD4+CD25+ Treg cells, directly induced CD4+CD45ROhi+ apoptosis. Our results provide direct evidence that Pam3CSK4 induces an immunomodulatory effect by inducing CD8+ Treg cells; therefore, it may be a good adjuvant for the treatment of mite allergies.


Comparative study of valved and nonvalved fully implantable catheters inserted via ultrasound-guided puncture for chemotherapy.
Fully implantable catheters are important for oncology treatments. They can be functionally categorized as valved or nonvalved. Theoretically, a valve prevents spontaneous blood reflux into the catheter, reducing the incidence of complications. We sought to compare the results from the implantation of valved and nonvalved fully implantable 8-French catheters inserted via ultrasound-guided puncture in cancer patients undergoing chemotherapy treatment. A retrospective analysis of 100 patients who underwent long-term catheter implantation guided by ultrasound was performed looking for early (≤30 days) or late (≥90 days) complications. They were evaluated regarding the use of valved or nonvalved catheter and assessed on univariate model. The only early complication (hematoma) was identified in the valved group. Twenty-two late complications were identified (72.72% in the valved group and 27.27% in the nonvalved group; P = 0.009). Blood reflux dysfunction, which occurred in 12 patients in the valved and none in the nonvalved devices group, was the only complication with an incidence that was significantly different between the groups. Fully implantable valved catheters with Groshong (Bard Medical Division, Covington, GA) valves have a higher rate of blood reflux dysfunction compared with nonvalved catheters, but this did not interfere with the efficacy of the treatment.


Genetic Diversity of Nitrogen-Fixing and Plant Growth Promoting Pseudomonas Species Isolated from Sugarcane Rhizosphere.
The study was designed to isolate and characterize Pseudomonas spp. from sugarcane rhizosphere, and to evaluate their plant- growth- promoting (PGP) traits and nitrogenase activity. A biological nitrogen-fixing microbe has great potential to replace chemical fertilizers and be used as a targeted biofertilizer in a plant. A total of 100 isolates from sugarcane rhizosphere, belonging to different species, were isolated; from these, 30 isolates were selected on the basis of preliminary screening, for in vitro antagonistic activities against sugarcane pathogens and for various PGP traits, as well as nitrogenase activity. The production of IAA varied from 312.07 to 13.12 μg mL-1 in tryptophan supplemented medium, with higher production in AN15 and lower in CN20 strain. The estimation of ACC deaminase activity, strains CY4 and BA2 produced maximum and minimum activity of 77.0 and 15.13 μmoL mg-1 h-1. For nitrogenase activity among the studied strains, CoA6 fixed higher and AY1 fixed lower in amounts (108.30 and 6.16 μmoL C2H2 h-1 mL-1). All the strains were identified on the basis of 16S rRNA gene sequencing, and the phylogenetic diversity of the strains was analyzed. The results identified all strains as being similar to Pseudomonas spp. Polymerase chain reaction (PCR) amplification of nifH and antibiotic genes was suggestive that the amplified strains had the capability to fix nitrogen and possessed biocontrol activities. Genotypic comparisons of the strains were determined by BOX, ERIC, and REP PCR profile analysis. Out of all the screened isolates, CY4 (Pseudomonas koreensis) and CN11 (Pseudomonas entomophila) showed the most prominent PGP traits, as well as nitrogenase activity. Therefore, only these two strains were selected for further studies; Biolog profiling; colonization through green fluorescent protein (GFP)-tagged bacteria; and nifH gene expression using quantitative real-time polymerase chain reaction (qRT-PCR) analysis. The Biolog phenotypic profiling, which comprised utilization of C and N sources, and tolerance to osmolytes and pH, revealed the metabolic versatility of the selected strains. The colonization ability of the selected strains was evaluated by genetically tagging them with a constitutively expressing GFP-pPROBE-pTetr-OT plasmid. qRT-PCR results showed that both strains had the ability to express the nifH gene at 90 and 120 days, as compared to a control, in both sugarcane varieties GT11 and GXB9. Therefore, our isolated strains, P. koreensis and P. entomophila may be used as inoculums or in biofertilizer production for enhancing growth and nutrients, as well as for improving nitrogen levels, in sugarcane and other crops. The present study, to the best of our knowledge, is the first report on the diversity of Pseudomonas spp. associated with sugarcane in Guangxi, China.


A novel PAAD-containing protein that modulates NF-kappa B induction by cytokines tumor necrosis factor-alpha and interleukin-1beta.
PAAD domains are found in diverse proteins of unknown function and are structurally related to a superfamily of protein interaction modules that includes death domains, death effector domains, and Caspase activation and recruitment domains. Using bioinformatics strategies, cDNAs were identified that encode a novel protein of 110 kDa containing a PAAD domain followed by a putative nucleotide-binding (NACHT) domain and several leucine-rich repeat domains. This protein thus resembles Cryopyrin, a protein implicated in hereditary hyperinflammation syndromes, and was termed PAN2 for PAAD and NACHT-containing protein 2. When expressed in HEK293 cells, PAN2 suppressed NF-kappaB induction by the cytokines tumor necrosis factor-alpha (TNFalpha) and interleukin-1beta (IL-1beta), suggesting that this protein operates at a point of convergence in these two cytokine signaling pathways. This PAN2-mediated suppression of NF-kappaB was evident both in reporter gene assays that measured NF-kappaB transcriptional activity and electromobility shift assays that measured NF-kappaB DNA binding activity. PAN2 also suppressed NF-kappaB induction resulting from overexpression of several adapter proteins and protein kinases involved in the TNF or IL-1 receptor signal transduction, including TRAF2, TRAF6, RIP, IRAK2, and NF-kappaB-inducing kinase as well as the IkappaB kinases IKKalpha and IKKbeta. PAN2 also inhibited the cytokine-mediated activation of IKKalpha and IKKbeta as measured by in vitro kinase assays. Furthermore, PAN2 association with IKKalpha was demonstrated by co-immunoprecipitation assays, suggesting a direct effect on the IKK complex. These observations suggest a role for PAN2 in modulating NF-kappaB activity in cells, thus providing the insights into the potential functions of PAAD family proteins and their roles in controlling inflammatory responses.


Recurrence of lymphoedema-associated cellulitis (erysipelas) under prophylactic antibiotherapy: a retrospective cohort study.
To identify the predictors of successful antibiotic prophylactic treatment using benzathin-penicillin G to prevent recurrence of erysipelas in patients with secondary upper limb lymphoedema. A retrospective cohort study. Patients with secondary arm lymphoedema were recruited in a single lymphology unit between 1990 and 2003. All patients had had at least three recurrences of erysipelas. Patients were given 2.4 MU benzathin-penicillin G intramuscularly at 14-day intervals. For each patient, the following data were recorded: characteristics of breast cancer treatment (type of surgery, radiotherapy, hormone therapy), number of erysipelas recurrences before inclusion, patient characteristics including body mass index (BMI) and lymphoedema volume at inclusion. The outcome studied was the occurrence of erysipelas on the affected arm under antibiotic prophylactic treatment. With a 4.2-year median follow-up from the onset of antibiotic prophylactic treatment, 23 of 48 women experienced recurrence of erysipelas. The median duration of erysipelas recurrence-free period under this treatment was 2.7 years. The estimated rate of recurrence was 26%[95% confidence interval (CI) 13-38%] at 1 year and 36% (95% CI 22-50%) at 2 years. No patient stopped the treatment because of side-effects. No predictive factor of erysipelas recurrence under antibiotic prophylactic treatment was identified. Antibiotic prophylaxis using benzathin-penicillin is a well-tolerated treatment of erysipelas recurrence in patients with upper limb lymphoedema secondary to breast cancer. The rate of erysipelas recurrence was 26% at 1 year in patients who had a history of at least one erysipelas. We did not find any predictor of erysipelas recurrence.


Learning Orthographic Structure With Sequential Generative Neural Networks.
Learning the structure of event sequences is a ubiquitous problem in cognition and particularly in language. One possible solution is to learn a probabilistic generative model of sequences that allows making predictions about upcoming events. Though appealing from a neurobiological standpoint, this approach is typically not pursued in connectionist modeling. Here, we investigated a sequential version of the restricted Boltzmann machine (RBM), a stochastic recurrent neural network that extracts high-order structure from sensory data through unsupervised generative learning and can encode contextual information in the form of internal, distributed representations. We assessed whether this type of network can extract the orthographic structure of English monosyllables by learning a generative model of the letter sequences forming a word training corpus. We show that the network learned an accurate probabilistic model of English graphotactics, which can be used to make predictions about the letter following a given context as well as to autonomously generate high-quality pseudowords. The model was compared to an extended version of simple recurrent networks, augmented with a stochastic process that allows autonomous generation of sequences, and to non-connectionist probabilistic models (n-grams and hidden Markov models). We conclude that sequential RBMs and stochastic simple recurrent networks are promising candidates for modeling cognition in the temporal domain.


Regulation of in vitro and in vivo immune functions by the cytosolic adaptor protein SKAP-HOM.
SKAP-HOM is a cytosolic adaptor protein representing a specific substrate for the Src family protein tyrosine kinase Fyn. Previously, several groups have provided experimental evidence that SKAP-HOM (most likely in cooperation with the cytosolic adaptor protein ADAP) is involved in regulating leukocyte adhesion. To further assess the physiological role of SKAP-HOM, we investigated the immune system of SKAP-HOM-deficient mice. Our data show that T-cell responses towards a variety of stimuli are unaffected in the absence of SKAP-HOM. Similarly, B-cell receptor (BCR)-mediated total tyrosine phosphorylation and phosphorylation of Erk, p38, and JNK, as well as immunoreceptor-mediated Ca(2+) responses, are normal in SKAP-HOM(-/-) animals. However, despite apparently normal membrane-proximal signaling events, BCR-mediated proliferation is strongly attenuated in the absence of SKAP-HOM(-/-). In addition, adhesion of activated B cells to fibronectin (a ligand for beta1 integrins) as well as to ICAM-1 (a ligand for beta2 integrins) is strongly reduced. In vivo, the loss of SKAP-HOM results in a less severe clinical course of experimental autoimmune encephalomyelitis following immunization of mice with the encephalitogenic peptide of MOG (myelin oligodendrocyte glycoprotein). This is accompanied by strongly reduced serum levels of MOG-specific antibodies and lower MOG-specific T-cell responses. In summary, our data suggest that SKAP-HOM is required for proper activation of the immune system, likely by regulating the cross-talk between immunoreceptors and integrins.


Fidelity scorecard: evaluation of a caregiver-delivered symptom management intervention.
To evaluate and quantify the intervention fidelity of a symptom management protocol through implementation of a scorecard, using an exemplar study of caregiver-delivered reflexology for people with breast cancer. Studies on caregiver-delivered symptom management interventions seldom include adequate information on protocol fidelity, contributing to potentially suboptimal provision of the therapeutic intervention, hindering reproducibility and generalizability of the results. Fidelity assessment of a 4-week intervention protocol in a randomized controlled trial (RCT) with data collection between 2012 - 2016. The National Institutes of Health Behaviour Change Consortium (NIH-BCC) conceptual model for intervention fidelity guided the study. The five NIH-BCC fidelity elements are: (1) dose; (2) provider training; (3) intervention delivery; (4) intervention receipt; and (5) enactment. To illustrate the elements, an intervention protocol was deconstructed and each element quantified using a newly developed fidelity scorecard. Mean scores and frequency distributions were derived for the scorecard elements. For dose, the mean number of sessions was 4·4, 96% used the correct intervention duration and 29% had 4 weeks with at least one session. Provider training was achieved at 80% of the maximum score, intervention delivery was 96%, intervention receipt was 99% and enactment indicated moderate adoption at 3·8 sessions per patient. The sample mean score was 15·4 out of 16, indicating the high overall fidelity. Research findings that include description of how fidelity is both addressed and evaluated are necessary for clinical translation. Clinicians can confidently recommend symptom management strategies to patients and caregivers when fidelity standards are explicitly reported and measured.


Public reporting and transparency: a primer on public outcomes reporting.
Healthcare consumers seeking accurate information about where to find quality surgical care face a confusing constellation of rating systems that lack transparency or consistency of opinion. For example, a 2016 report in Health Affairs demonstrated that no hospital was rated as a high performer by all four prominent national ratings systems: Consumer Reports, Leapfrog, Healthgrades and U.S. News & World Report (Austin et al. Health Aff 34:423-430, 2015). Surgeons should have an understanding of the current state of public reporting of quality; hospital ratings and data sources; physician ratings and data sources; and transparency of reporting. We conducted a non-systematic review of the literature. Hospital quality ratings remain nebulous and there is not universal opinion on the utility of voluntary participation in ranking systems, leaving the current systems largely opinion-based. Early attempts at physician ranking systems are rudimentary at best and suffer from methodological concerns. Publicly reported metrics should be easily understandable, accessible, clinically relevant, reliable, non-punitive, and shielded from legal discovery. Transparency is increasing within institutions to help align staff to institutional objectives, while specialty specific registries are helping to standardize care pathways and outcomes measures across organizations. Measuring surgical outcomes beyond 30-day morbidity and mortality has been plagued by a lack of understanding on how to create metrics that matter; the four attributes of relevance, scientific soundness, feasibility and comprehensiveness set a high bar for the development of effective and efficient quality measures in surgery. SAGES, via the Quality, Outcomes, and Safety Committee, is committed to learning how to develop meaningful quality metrics in general surgery and will continue to work in other areas that impact quality, such as opioid prescribing, and surgeon wellness.


Lateral Habenula Mediates Defensive Responses Only When Threat and Safety Memories Are in Conflict.
Survival depends on the ability to adaptively react or execute actions based on previous aversive salient experiences. Although lateral habenula (LHb) activity has been broadly implicated in the regulation of aversively motivated responses, it is not clear under which conditions this brain structure is necessary to regulate defensive responses to a threat. To address this issue, we combined pharmacological inactivations with behavioral tasks that involve aversive and appetitive events and evaluated defensive responses in rats. We found that LHb pharmacological inactivation did not affect cued threat conditioning (fear) and extinction (safety) learning and memory, anxiety-like or reward-seeking behaviors. Surprisingly, we found that LHb inactivation abolished reactive defensive responses (tone-elicited freezing) only when threat (conditioning) and safety memories (extinction and latent inhibition) compete during retrieval. Consistently, we found that LHb inactivation impaired active defensive responses [platform-mediated avoidance (PMA)], thereby biasing choice behavior (between avoiding a threat or approaching food) toward reward-seeking responses. Together, our findings suggest that LHb activity mediates defensive responses only when guided by competing threat and safety memories, consequently revealing a previously uncharacterized role for LHb in experience-dependent emotional conflict.


Complete genome sequence of hyperthermophilic Pyrococcus sp. strain NA2, isolated from a deep-sea hydrothermal vent area.
Pyrococcus sp. strain NA2, isolated from a deep-sea hydrothermal vent sample, is a novel marine hyperthermophilic archaeon that grows optimally at 93 °C. The complete genome sequence of the strain contains all the genes for the tricarboxylic acid cycle except for succinate dehydrogenase/fumarate reductase, but the genome does not encode proteins involved in polysaccharide utilization.


Short course dexamethasone treatment following injury inhibits bleomycin induced fibrosis in rats.
Corticosteroids are routinely used in patients with pulmonary fibrosis. The timing for initiation of treatment is likely to be crucial for corticosteroids to exert an antifibrotic effect. Experimental studies in animals have examined the effect of corticosteroid treatment starting before or at the time of lung injury. However, this is not representative of the human condition as treatment only begins after disease has been established. We examined the effect of a short course corticosteroid treatment starting 3 days after bleomycin induced lung injury on the development of pulmonary fibrosis. Bleomycin (1.5 mg/kg) was instilled intratracheally into rats to induce pulmonary fibrosis. The effect of a 3-day course of dexamethasone (0.5 mg/kg) initiated 3 days after bleomycin induced lung injury on cell proliferation and collagen deposition was examined by analysing bronchoalveolar lavage (BAL) fluid and lung tissue. Treating bleomycin exposed animals after injury with dexamethasone for 3 days inhibited lung collagen deposition compared with animals exposed to bleomycin without dexamethasone treatment (15.2 (2.2) mg collagen/lung v 22.5 (2.1) mg/lung; p<0.05). Dexamethasone treatment reduced pulmonary parenchymal cell proliferation in bleomycin exposed rats but did not influence BAL fluid mitogenic activity for lung fibroblasts or alter the BAL fluid levels of the fibrogenic mediators transforming growth factor-beta(1), platelet derived growth factor-AB, and thrombin. A 3 day course of dexamethasone treatment initiated 3 days after bleomycin induced lung injury reduces lung cell proliferation and collagen deposition by mechanisms other than through reduction of transforming growth factor-beta(1), platelet derived growth factor-AB, and thrombin levels in BAL fluid. We propose that an early short course treatment with dexamethasone may be useful in inhibiting pulmonary fibrosis.


From Hong Kong Diabetes Register to JADE Program to RAMP-DM for Data-Driven Actions.
In 1995, the Hong Kong Diabetes Register (HKDR) was established by a doctor-nurse team at a university-affiliated, publicly funded, hospital-based diabetes center using a structured protocol for gathering data to stratify risk, triage care, empower patients, and individualize treatment. This research-driven quality improvement program has motivated the introduction of a territory-wide diabetes risk assessment and management program provided by 18 hospital-based diabetes centers since 2000. By linking the data-rich HKDR to the territory-wide electronic medical record, risk equations were developed and validated to predict clinical outcomes. In 2007, the HKDR protocol was digitalized to establish the web-based Joint Asia Diabetes Evaluation (JADE) Program complete with risk levels and algorithms for issuance of personalized reports to reduce clinical inertia and empower self-management. Through this technologically assisted, integrated diabetes care program, we have generated big data to track secular trends, identify unmet needs, and verify interventions in a naturalistic environment. In 2009, the JADE Program was adapted to form the Risk Assessment and Management Program for Diabetes Mellitus (RAMP-DM) in the publicly funded primary care clinics, which reduced all major events by 30-60% in patients without complications. Meanwhile, a JADE-assisted assessment and empowerment program provided by a university-affiliated, self-funded, nurse-coordinated diabetes center, aimed at complementing medical care in the community, also reduced all major events by 30-50% in patients with different risk levels. By combining universal health coverage, public-private partnerships, and data-driven integrated care, the Hong Kong experience provides a possible solution than can be adapted elsewhere to make quality diabetes care accessible, affordable, and sustainable.


Lead exposure from backyard chicken eggs: a public health risk?
Although the USA has made significant strides in reducing lead exposure, new and emerging sources are raising cause for public concern. Recent reports of finding lead in eggs from chickens raised in urban gardens has highlighted the need to consider the potential health risks of consuming eggs from backyard chickens. Following the detection of 0.33 μg/g lead in the edible portion of eggs submitted for lead analysis from a backyard chicken owner, further investigation was conducted to determine the source and extent of lead exposure in the flock. Several birds, almost two dozen eggs, and environmental samples were submitted to the California Animal Health and Food Safety Laboratory for further testing. Lead was detected in the blood, liver, kidney, and bone at varying concentrations in all birds but was not detected in the muscle tissue. All egg shells contained detectable amounts of lead, while only a little over half of the edible portion of the eggs contained lead. The detected concentrations in the edible portion approached or exceeded the recommended threshold of lead consumption per day that should not be exceeded by young children if a child consumed one average-sized egg. Peeling paint from a wooded structure adjacent to the flock's coop was the likely lead source containing 3,700 μg/g lead. Thus, removal of the chickens from the source and periodic testing of eggs for lead were recommended. This case illustrates the need for consumers and health care workers to be aware of potential sources for lead exposure such as backyard chickens.


A case of treatment of multidrug-resistant intracranial Klebsiella pneumoniae infection by multichannel colistin sulfate.
The management of severe neurologic infections due to multidrug-resistant (MDR) Klebsiella pneumoniae infection remains a challenge. Limited antibiotic treatment regimens make treatment of severe MDR K. pneumoniae infection more difficult. We describe a patient who developed severe meningitis and ventriculitis after craniotomy caused by MDR K. pneumoniae and was effectively treated with the administration of multichannel applications (intravenous, intrathecal and aerosol inhalation) of colistin sulfate. This case provides clinical evidence that the intrathecal, intravenous and aerosol inhalation of colistin sulfate by multichannel application can be a last resort in refractory intracranial infection by MDR K. pneumoniae.


Low volume ventilation with permissive hypercapnia in the Adult Respiratory Distress Syndrome.
Many animal studies have demonstrated that mechanical ventilation with high peak inspiratory pressures (PIP) can result in a form of acute lung injury closely resembling ARDS, ie characterised by hyaline membranes, granulocyte infiltration, increased pulmonary and systemic vascular permeability, and eventually proliferation of fibroblasts and type II pneumocytes. These studies have led to a concern that, in some patients, orthodox ventilatory management in severe ARDS may result in additional lung injury and, possibly, remote organ dysfunction. Mortality may be increased as a consequence. In an attempt to avoid such ventilator-induced lung injury in severe ARDS, several modifications of ventilatory management have been evaluated. We have previously reported the technique of low volume pressure limited ventilation with permissive hypercapnia, using tidal volumes of 5-7 ml/kg and allowing the PaCO 2 to rise substantially (maximum PaCO 2 17.2 kPa [129 mmHg]), mean maximum 8.3 kPa [62 mmHg]). In an uncontrolled study the mortality was significantly lower than that predicted by Apache II (16% vs 39.6%, p less than 0.01). Acute hypercapnia can cause many physiological disturbances but most of these appear to be due to the resulting intracellular acidosis and should not occur in hypercapnia of gradual onset, allowing the intracellular pH to be normalised. The time scales for compensation of intracellular and extracellular acidosis are markedly different. However, even severe acute hypercapnia appears to be remarkably well tolerated. Several clinical studies suggest that the avoidance of high PIP may reduce mortality in ARDS, but a randomised trial will be required to establish whether pressure limitation and permissive hypercapnia do improve outcome.


Ultrasonographic study of the temporomandibular joint in individuals with and without temporomandibular disorder.
The present study's aim was to compare temporomandibular joint (TMJ) images for individuals with and without temporomandibular disorder (TMD) using high-resolution ultrasonography (HRUS). The distance between the lateral-most point of the articular capsule and the lateral-most point of the mandibular condyle (lateral capsule-mandibular condyle distance) was determined to confirm the clinical diagnosis according to the Diagnostic Criteria for Temporomandibular Disorders (DC/TMD). The sample group comprised 17 women and 15 men, aged 19-39 years, distributed into TMD (n = 20) and Asymptomatic (n = 12) groups. The evaluations were in closed- and open-mouth positions. The lateral capsule-mandibular condyle distance was measured in both positions. The articular surface of the condyle and the articular capsule were visualized as hyperechoic structures, and the articular disk was visualized as a central, hyperechogenic area surrounded by a hypoechoic linear image. Morphological changes were observed in some TMD group participants, including joint effusion (hypoechogenic area), condylar erosions (increased hyperechoic area), and condylar surface irregularities. The lateral capsule-mandibular condyle distance did not differ between sides or groups among participants with intra-articular disorders with or without pain (P > 0.05). In conclusion, HRUS allowed visualization of the TMJ structures, but did not allow confirmation of clinical diagnosis by DC/TMD.


Oral chronic graft-versus-host disease: analysis of dendritic cells subpopulations.
The graft-versus-host disease is the major cause of morbidity and mortality in patients who have undergone hematopoietic stem cell transplantation. Aiming at contributing to the understanding of the role of myeloid and plasmacytoid dendritic cells, and natural killer cells in chronic graft-versus-host disease, we examined biopsies of jugal mucosa of 26 patients with acute myeloid leukemia who had undergone allogenic hematopoietic stem cell transplantation. Half of these patients developed oral chronic graft-versus-host disease. Microscopic sections were immunohistochemically stained for anti-CD1a, anti-CD123 and anti-CD56. We calculated the number of immunostained cells in the corium per square millimeter and applied the Mann-Whitney test. Results showed a statistically significant increase of myeloid dendritic cells (CD1a+; p=0,02) and natural killer cells (CD56; p=0,04) in patients with oral chronic graft-versus-host disease. CD123 immunostaining showed no statistical difference between groups. It was concluded that myeloid dendritic cells and natural killer cells participate in the development of oral chronic graft-versus-host disease.


Spontaneous growth of 2D coordination polymers on functionalized ferromagnetic surfaces.
The spontaneous growth of lamellar metal-alkanethiolates (LMAs) on reactive ferromagnetic surfaces as a result of surface oxidation has been observed. When alkanethiol self-assembled monolayers (SAMs) grown under an inert atmosphere over cobalt or permalloy (Ni80Fe20) are exposed to air, oxygen diffuses through the molecular layer. This induces an oxidation of metal atoms at the metal surface and a release of the resulting metal cations that migrate coordinated by the alkanethiol molecules to form lamellar structures over the SAMs. This process has been imaged in real-time, under ambient conditions, by means of different microscopy techniques. The influence of the alkyl chain length, the nature of the ferromagnet, the temperature and the atmospheric moisture on the number, area and height of the resulting features has been systematically evaluated. Remarkably, the possibility to follow the migration in real-time makes it a promising model system for the study of surface/molecule interface processes. Most importantly, the composition and crystallinity of these LMAs have been studied, evidencing that real 2D coordination polymers are formed on the surface. Hence, one could envision this strategy as a new method for the assembly of more complex low-dimensional (2D) magnetic materials based on coordination polymers.


Comprehensive analysis of expression profile and prognostic significance of interferon regulatory factors in pancreatic cancer.
Pancreatic cancer (PC) is a highly lethal disease and an increasing cause of cancer-associated mortality worldwide. Interferon regulatory factors (IRFs) play vital roles in immune response and tumor cellular biological processes. However, the specific functions of IRFs in PC and tumor immune response are far from systematically clarified. This study aimed to explorer the expression profile, prognostic significance, and biological function of IRFs in PC. We observed that the levels of IRF2, 6, 7, 8, and 9 were elevated in tumor compared to normal tissues in PC. IRF7 expression was significantly associated with patients' pathology stage in PC. PC patients with high IRF2, low IRF3, and high IRF6 levels had significantly poorer overall survival. High mRNA expression, amplification and, deep deletion were the three most common types of genetic alterations of IRFs in PC. Low expression of IRF2, 4, 5, and 8 was resistant to most of the drugs or small molecules from Genomics of Drug Sensitivity in Cancer. Moreover, IRFs were positively correlated with the abundance of tumor infiltrating immune cells in PC, including B cells, CD8+ T cells, CD4+ T cells, macrophages, Neutrophil, and Dendritic cells. Functional analysis indicated that IRFs were involved in T cell receptor signaling pathway, immune response, and Toll-like receptor signaling pathway. Our results indicated that certain IRFs could serve as potential therapeutic targets and prognostic biomarkers for PC patients. Further basic and clinical studies are needed to validate our findings and generalize the clinical application of IRFs in PC.


[Specificity of biochemical markers of pregnancy second trimester].
During the second trimester of 21 trisomy pregnancies, median of maternal serum concentrations of alpha-fetoprotein, not conjugated estriol, beta fraction, and inhibin A differ from normal values, and it can be used to select high risk patients. To evaluate specificity of different forms of second trimester biochemical markers association in a group of healthy pregnant women and determine levels considered as low risk. Random and cross-sectional study in 363 pregnant women with prenatal diagnosis to evaluate specificity of different combinations of biochemical markers. Down's syndrome and defects of neural tube rates of detection were defined as the percentage of pregnancies with tests of detection of true negatives and false positives. Total beta fraction has differences during weeks 13 to 14, with 70% of cases between 23,000 to 70,000 units, with 1.38% of false positives; alpha-fetoprotein fluctuated between 22 to 30 units, with 5.2% of false positives; conjugated estriol increases between 1.2 to 2.2 units during 2nd quarterly, with 6.3% of false positives; and inhibin has an increase, with 70% of cases located between 125 to 486.5 units, with 10.44% of false positives. Marker formula with greater specificity was type IV, included alpha-fetoprotein, not conjugated estriol and beta fraction. In the isolated form, alpha-fetoprotein marker demonstrates greater specificity.


A peripheral and central T cell antigen recognized by a monoclonal thymocytotoxic autoantibody from New Zealand black mice.
Naturally occurring thymocytotoxic autoantibodies (NTA) have been suggested to be the cause of thymic atrophy and T cell disorders in human and murine lupus. Definitive studies on NTA's role in the induction of SLE, however, have been lacking due to the lack of a pure source of NTA. Although it is clear that NTA are a heterogeneous group of antibodies, the nature of their antigens has remained obscure. We report the characteristics of a monoclonal NTA, designated SAG-3, which appears more reflective of the activities previously reported of serum NTA than other NTA-secreting clones. SAG-3 is an IgM autoantibody cytotoxic for 80 to 90% of thymocytes, 20 to 25% of splenic lymphocytes, 25 to 30% of lymph node cells, and less than 3% cortisol-resistant thymocytes, bone marrow, and fetal liver cells. SAG-3 is murine-specific without reactivity towards rat, hamster, or guinea pig, and appears very early in thymic development, on day 17 fetal thymocytes. SAG-3 is equally cytotoxic against several strains of mice, including both Thy-1.1 and Thy-1.2 allotypes, and the cytotoxicity is absorbed by brain but not liver cells. Reactive thymocytes occurred throughout the cortical regions of the thymus, indicating preferential affinity towards immature thymocytes. Although the serologic activities of SAG-3 suggest that Thy-1 alloantigen is its target, SAG-3 antigen is found to be distinct from Thy-1 and also from Lyt-1, Lyt-2, or L3T4 antigens. The binding of SAG-3 to thymocytes could be competitively inhibited by NTA-positive NZB sera.


Resident Burnout and Work Environment.
Objective We examined the prevalence of burnout among resident doctors and its relationship with specific stressors. Method We conducted a nationwide, online, cross-sectional survey in Japan with 604 resident doctors in 2018-2019. Materials Participants completed the Maslach Burnout Inventory-General Survey to evaluate burnout and provided details of their individual factors and working environmental factors. Chi-square tests and t-tests were conducted for categorical and continuous variables, respectively. The association between burnout and resident-reported causes of stress, ways of coping with stress, number of times patient-safety incidents were likely to occur, and individuals who provide support when in trouble was analyzed using logistic regression analyses after controlling for confounding variables. Results A total of 28% met the burnout criteria, 12.2% were exhausted, 2.8% were depressed, and 56.9% were healthy. After adjusting for sex, postgraduate years, type of residency program, marital status, number of inpatients under residents' care, number of working hours, number of night shifts, number of days off, and resident-reported causes of stress - excessive paperwork [odds ratio (OR): 2.24, 95% confidence interval (CI): 1.32-3.80], excessive working hours (OR: 2.75, 95% CI: 1.24-6.04), low autonomy (OR: 3.92, 95% CI: 2.01-7.65), communication problems at the workplace (OR: 2.24, 95% CI: 1.05-4.76), complaints from patients (OR: 6.62, 95% CI: 1.21-36.1), peer competition (OR: 2.22, 95% CI: 1.25-3.93), and anxiety about the future (OR: 2.13, 95% CI: 1.28-3.56) - were independently associated with burnout. The burnout group had more reported patient-safety incidents that were likely to occur per year (>10) (OR: 2.65, 95% CI: 1.01-6.95) and a lack of individuals who could provide support when in trouble (OR: 1.83, 95% CI: 1.01-3.34) than the non-burnout group. Conclusion This study described the prevalence of burnout among residents who responded to our survey. We detected an association between burnout and resident-reported causes of stress, patient-safety incidents, and a lack of individuals who provide support when in trouble. Further interventional studies targeting ways to reduce these concerns are warranted.


Physical growth of cebus monkeys, Cebus albifrons, during protein or calorie deficiency.
Infant cebus monkey (Cebus albifrons) on experimental diets providing low-protein (2.8% of calories) or low-calorie (67% of ad libitum intake) levels for 20 weeks beginning at 8 weeks of age showed marked changes in their patterns of physical growth. Significant size differences between the malnourished animals and the control group appeared within 4 weeks. Although rates of growth were significantly reduced in all measurements, all of the malnourished monkeys, including low-protein animals showing zero weight gain, continued skeletal growth (except in skull measurements) at low levels for the duration of experiment. Both the protein- and calorie-restricted animals developed a thin, emaciated appearance often associated with marasmus, not by the continuous loss of tissue byt by the redistribution of the tissue over a slowly expanding skeleton. For many skeletal proportions, the pattern and shape development in the malnourished animals differed from that of the control animals. Growth during malnutrition was most affected in those parts of the skeleton which were more advanced in relative size.


Functional domains of the Bacillus subtilis transcription factor AraR and identification of amino acids important for nucleoprotein complex assembly and effector binding.
The Bacillus subtilis AraR transcription factor represses at least 13 genes required for the extracellular degradation of arabinose-containing polysaccharides, transport of arabinose, arabinose oligomers, xylose, and galactose, intracellular degradation of arabinose oligomers, and further catabolism of this sugar. AraR exhibits a chimeric organization comprising a small N-terminal DNA-binding domain that contains a winged helix-turn-helix motif similar to that seen with the GntR family and a larger C-terminal domain homologous to that of the LacI/GalR family. Here, a model for AraR was derived based on the known crystal structures of the FadR and PurR regulators from Escherichia coli. We have used random mutagenesis, deletion, and construction of chimeric LexA-AraR fusion proteins to map the functional domains of AraR required for DNA binding, dimerization, and effector binding. Moreover, predictions for the functional role of specific residues were tested by site-directed mutagenesis. In vivo analysis identified particular amino acids required for dimer assembly, formation of the nucleoprotein complex, and composition of the sugar-binding cleft. This work presents a structural framework for the function of AraR and provides insight into the mechanistic mode of action of this modular repressor.


High-Temperature Phase Transition and Dissociation of (Mg, Fe)SiO3 Perovskite at Lower Mantle Pressures.
To study the crystallography of Earth's lower mantle, techniques for measuring synchrotron x-ray diffraction from a laser-heated diamond anvil cell have been developed. Experiments on samples of (Mg, Fe)SiO(3) show that silicate perovskite maintains its orthorhombic symmetry at 38 gigapascals and 1850 kelvin. Measurements at 65 and 70 gigapascals provide evidence for a temperature-induced orthorhombic-to-cubic phase transition and dissociation to an assemblage of perovskite and mixed oxides. If these phase transitions occur in Earth, they will require a significant change in mineralogical models of the lower mantle.


Establishing a probabilistic reversal learning test in mice: evidence for the processes mediating reward-stay and punishment-shift behaviour and for their modulation by serotonin.
Valid animal models of psychopathology need to include behavioural readouts informed by human findings. In the probabilistic reversal learning (PRL) task, human subjects are confronted with serial reversal of the contingency between two operant stimuli and reward/punishment and, superimposed on this, a low probability (0.2) of punished correct responses/rewarded incorrect responses. In depression, reward-stay and reversals completed are unaffected but response-shift following punished correct response trials, referred to as negative feedback sensitivity (NFS), is increased. The aims of this study were to: establish an operant spatial PRL test appropriate for mice; obtain evidence for the processes mediating reward-stay and punishment-shift responding; and assess effects thereon of genetically- and pharmacologically-altered serotonin (5-HT) function. The study was conducted with wildtype (WT) and heterozygous mutant (HET) mice from a 5-HT transporter (5-HTT) null mutant strain. Mice were mildly food deprived and reward was sugar pellet and punishment was 5-s time out. Mice exhibited high motivation and adaptive reversal performance. Increased probability of punished correct response (PCR) trials per session (p = 0.1, 0.2 or 0.3) led to monotonic decrease in reward-stay and reversals completed, suggesting accurate reward prediction. NFS differed from chance-level at p PCR = 0.1, suggesting accurate punishment prediction, whereas NFS was at chance-level at p = 0.2-0.3. At p PCR = 0.1, HET mice exhibited lower NFS than WT mice. The 5-HTT blocker escitalopram was studied acutely at p PCR = 0.2: a low dose (0.5-1.5 mg/kg) resulted in decreased NFS, increased reward-stay and increased reversals completed, and similarly in WT and HET mice. This study demonstrates that testing PRL in mice can provide evidence on the regulation of reward and punishment processing that is, albeit within certain limits, of relevance to human emotional-cognitive processing, its dysfunction and treatment.


Early carotid artery stenting and angioplasty in patients with acute ischemic stroke.
To determine the safety of early percutaneous endovascular carotid angioplasty and stenting (CAS) after an ischemic stroke. The neurointerventional database was reviewed for patients who underwent CAS after an acute ischemic stroke in two university hospitals. Clinical and radiological data were reviewed. Outcomes were worsening stroke, new stroke, or stroke-related death up to 30 days after the procedure. Procedure-related complications were also documented. A total of 38 patients with 39 procedures were identified. The mean age was 67 +/- 15 years; 31 of 38 patients were Caucasian and 24 were female. Hypertension was found in 21 patients, peripheral vascular disease in 12, diabetes in 13, and coronary artery disease in 18. The median initial National Institutes of Health Stroke Scale score was 8. The carotid artery showed severe to high-grade stenosis in 28 patients, dissection was present in 6, and the rest had an acute occlusion treated with thrombolysis followed by CAS. The mean time from stroke onset to CAS was 55 +/- 34 hours. The mean degree of stenosis at baseline was 86 +/- 11%. In 37 procedures, complete recanalization was achieved, defined as less than 10% residual narrowing; in 2 procedures, the residual stenosis was mild (10-20%). Neurological deterioration occurred after three procedures (7.7%), with minor nondisabling stroke in two and death from intracranial hemorrhage in one. If deemed necessary and in certain circumstances, early CAS seems to be safe after acute ischemic stroke if infarction volume is small and neurological deficit is mild.


Changes in the glucose and insulin responses according to high-protein snacks for diabetic patients.
This study aimed to develop healthy, appetizing high-protein snacks with enhanced isolated soy protein for diabetic patients and determine the blood glucose and insulin response after being consumed by these patients. Thirty adult patients aged between 30 and 75 years, with a ≤ 10-year history of type 2 diabetes and hemoglobin A1c of < 7.5%, were enrolled in this study. They made 3 clinical visits at one-week intervals. The control group consumed 50 g carbohydrates (white bread), whereas the test groups consumed high-protein grain (HP_G) or high-protein chocolate (HP_C) after an 8-hrs fast. Blood (2 cm3) was drawn at 15, 30, 45, 60, 90, and 120 min before and after consumption to analyze the blood glucose and insulin concentrations. Compared to the commercial snacks, the developed high-protein snacks had below-average calorie, carbohydrate, and fat content and a 2.5-fold higher protein content. In diabetic patients who consumed these snacks, the postprandial blood glucose increased between 15 min and 2 h after consumption, which was significantly slower than the time taken for the blood glucose to increase in the patients who consumed the control food product (P < 0.001). Insulin secretion was significantly lower at 45 min after consumption (P < 0.05), showing that the high-protein snacks did not increase the blood glucose levels rapidly. The incremental area under the curve (iAUC), which indicated the degree of blood sugar and insulin elevation after food intake, was higher in the control group than the groups given the 2 developed snacks (P < 0.001), and there was no significant difference in insulin secretion. The results of the postprandial blood glucose and insulin response suggest that high-protein snacks are potential convenient sources of high-quality protein and serve as a healthier alternative for patients with type 2 diabetes, who may have limited snack product choices. Such snacks may also provide balanced nutrition to pre-diabetic and obese individuals.


Heparin/heparan sulfate interacting protein expression and functions in human breast cancer cells and normal breast epithelia.
Heparin/heparan sulfate interacting protein (HIP) is a recently identified protein expressed by many normal epithelia and epithelial cell lines. In the present study, we examined expression and potential functions of this protein in a series of human breast cancer cells and in sections of normal and malignant human breast tissue. Four of the five breast cancer cell lines studied (MCF-7, T-47D, MDA-MB468, and BT-549) expressed HIP protein and mRNA at similar levels. In contrast, MDA-MB-231 cells failed to display reactivity with HIP-specific probes in any assay. Cell aggregation assays and cell surface antibody binding studies demonstrated that HIP was expressed on the cell surface. However, HIP expression did not correlate with the number of cell surface [3H]heparin (HP) binding sites. The K(Dapp)s for cell surface HP binding sites were similar in all breast cancer cell lines studied and ranged from 112 to 298 nM. In contrast, cell surface HP binding capacity varied greatly, ranging from 2.3 x 10(5) (MDA-MB-231 and MDA-MB-468) to 99 x 10(5) sites/cell (BT-549). All cell lines tested displayed the ability to bind to a heparan sulfate (HS)-binding synthetic peptide motif of HIP in a HP-inhibitable fashion. Binding to this motif was not inhibited by other glycosaminoglycans including hyaluronic acid, chondroitin sulfates, or keratan sulfate. Furthermore, cell binding to HIP peptide was almost completely lost when intact cells were predigested with heparinases but not chondroitinases. Cell surface HS from breast cancer cells as well as normal human breast epithelia binded to HIP peptide in a HP-inhibitable fashion, demonstrating the ability of these cell surface components to directly interact. HIP was detected in both normal breast epithelia and breast tumors in situ. It is suggested that HIP mediates aspects of HS-dependent interactions of both normal and malignant breast epithelia with other cells and extracellular matrix components.


Reactivity of Thiol-Rich Zn Sites in Diacylglycerol-Sensing PKC C1 Domain Probed by NMR Spectroscopy.
Conserved homology 1 (C1) domains are peripheral zinc finger domains that are responsible for recruiting their host signaling proteins, including Protein Kinase C (PKC) isoenzymes, to diacylglycerol-containing lipid membranes. In this work, we investigated the reactivity of the C1 structural zinc sites, using the cysteine-rich C1B regulatory region of the PKCα isoform as a paradigm. The choice of Cd2+ as a probe was prompted by previous findings that xenobiotic metal ions modulate PKC activity. Using solution NMR and UV-vis spectroscopy, we found that Cd2+ spontaneously replaced Zn2+ in both structural sites of the C1B domain, with the formation of all-Cd and mixed Zn/Cd protein species. The Cd2+ substitution for Zn2+ preserved the C1B fold and function, as probed by its ability to interact with a potent tumor-promoting agent. Both Cys3His metal-ion sites of C1B have higher affinity to Cd2+ than Zn2+, but are thermodynamically and kinetically inequivalent with respect to the metal ion replacement, despite the identical coordination spheres. We find that even in the presence of the oxygen-rich sites presented by the neighboring peripheral membrane-binding C2 domain, the thiol-rich sites can successfully compete for the available Cd2+. Our results indicate that Cd2+ can target the entire membrane-binding regulatory region of PKCs, and that the competition between the thiol- and oxygen-rich sites will likely determine the activation pattern of PKCs.


Selection Into Mental Health Services Among Persons With Depression.
This study aimed to identify sociodemographic and health characteristics associated with use of different mental health services (medication only, counseling only, or both) among persons with depression. The analytic sample consisted of adults who had a major depressive episode in the past year and received outpatient professional mental health services (N=4,169). Multinomial logistic regressions were computed with data from the 2015 and 2016 National Survey on Drug Use and Health to identify factors associated with the relative odds of receiving each modality of mental health service. Sixty-nine percent of the sample received both prescription medication and counseling (talking to a professional health care provider about depression), 22% received counseling only, and 9% received medication only. Being ordered into care and higher probability of having a severe mental illness were associated with higher odds of receiving both medication and counseling. How people with depression enter care and select into different mental health service modalities might be an indicator of access. Factors that affect selection into these modalities might also be associated with outcomes of care. Findings could inform efforts to remove modality-specific barriers to treatment, improve timely access to care, and reduce unmet need for mental health care among persons with depression.


Once-daily simeprevir (TMC435) with pegylated interferon and ribavirin in treatment-naïve genotype 1 hepatitis C: the randomized PILLAR study.
The phase IIb, double-blind, placebo-controlled PILLAR trial investigated the efficacy and safety of two different simeprevir (SMV) doses administered once-daily (QD) with pegylated interferon (Peg-IFN)-α-2a and ribavirin (RBV) in treatment-naïve patients with HCV genotype 1 infection. Patients were randomized to one of five treatments: SMV (75 or 150 mg QD) for 12 or 24 weeks or placebo, plus Peg-IFN and RBV. Patients in the SMV arms stopped all treatment at week 24 if response-guided therapy (RGT) criteria were met; patients not meeting RGT continued with Peg-IFN and RBV until week 48, as did patients in the placebo control group. Sustained virologic response (SVR) rates measured 24 weeks after the planned end of treatment (SVR24) were 74.7%-86.1% in the SMV groups versus 64.9% in the control group (P < 0.05 for all comparisons [SMV versus placebo], except SMV 75 mg for 24 weeks). Rapid virologic response (HCV RNA <25 IU/mL undetectable at week 4) was achieved by 68.0%-75.6% of SMV-treated and 5.2% of placebo control patients. According to RGT criteria, 79.2%-86.1% of SMV-treated patients completed treatment by week 24; 85.2%-95.6% of these subsequently achieved SVR24. The adverse event profile was generally similar across the SMV and placebo control groups, with the exception of mild reversible hyperbilirubinemia, without serum aminotransferase abnormalities, associated with higher doses of SMV. SMV QD in combination with Peg-IFN and RBV significantly improves SVR rates, compared with Peg-IFN and RBV alone, and allows the majority of patients to shorten their therapy duration to 24 weeks.


Raf and MEK protein kinases are direct molecular targets for the chemopreventive effect of quercetin, a major flavonol in red wine.
Considerable attention has focused on the health-promoting effects of red wine and its nonflavonoid polyphenol compound resveratrol. However, the underlying molecular mechanisms and molecular target(s) of red wine or other potentially active ingredients in red wine remain unknown. Here, we report that red wine extract (RWE) or the red wine flavonoid quercetin inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced transformation of JB6 promotion-sensitive mouse skin epidermal (JB6 P+) cells. The activation of activator protein-1 and nuclear factor-kappaB induced by TPA was dose dependently inhibited by RWE or quercetin treatment. Western blot and kinase assay data revealed that RWE or quercetin inhibited mitogen-activated protein kinase/extracellular signal-regulated kinase (ERK) kinase (MEK) 1 and Raf1 kinase activities and subsequently attenuated TPA-induced phosphorylation of ERK/p90 ribosomal S6 kinase. Although either RWE or quercetin suppressed Raf1 kinase activity, they were more effective in inhibiting MEK1 activity. Importantly, quercetin exerted stronger inhibitory effects than PD098059, a well-known pharmacologic inhibitor of MEK. Resveratrol did not affect either MEK1 or Raf1 kinase activity. Pull-down assays revealed that RWE or quercetin (but not resveratrol) bound with either MEK1 or Raf1. RWE or quercetin also dose dependently suppressed JB6 P+ cell transformation induced by epidermal growth factor or H-Ras, both of which are involved in the activation of MEK/ERK signaling. Docking data suggested that quercetin, but not resveratrol, formed a hydrogen bond with the backbone amide group of Ser(212), which is the key interaction for stabilizing the inactive conformation of the activation loop of MEK1.


Adherence to treatment in adolescents with cystic fibrosis: the role of illness perceptions and treatment beliefs.
This study was conducted to explore the relationships between illness perceptions, emotional representations, treatment beliefs and reported adherence in adolescents with cystic fibrosis (CF). Thirty-eight adolescents completed questionnaires assessing their perceptions of CF, beliefs about prescribed treatments and reported adherence to chest physiotherapy, enzyme supplements, and antibiotics. Reported non-adherence to chest physiotherapy was associated with the way in which patients judged their personal need for treatment relative to their concerns about potential adverse effects. Patients reported strong doubts about the necessity of chest physiotherapy. Reported non-adherence to antibiotics was related to doubts about the necessity of antibiotics, believing that CF is not amenable to treatment control. Despite these beliefs about treatment, participants perceived CF as a chronic condition. The findings provide preliminary support for the self-regulatory model, using the necessity-concerns framework to operationalize treatment beliefs, in explaining adherence to treatment in adolescents with CF.


Human and mouse mutations in WDR35 cause short-rib polydactyly syndromes due to abnormal ciliogenesis.
Defects in cilia formation and function result in a range of human skeletal and visceral abnormalities. Mutations in several genes have been identified to cause a proportion of these disorders, some of which display genetic (locus) heterogeneity. Mouse models are valuable for dissecting the function of these genes, as well as for more detailed analysis of the underlying developmental defects. The short-rib polydactyly (SRP) group of disorders are among the most severe human phenotypes caused by cilia dysfunction. We mapped the disease locus from two siblings affected by a severe form of SRP to 2p24, where we identified an in-frame homozygous deletion of exon 5 in WDR35. We subsequently found compound heterozygous missense and nonsense mutations in WDR35 in an independent second case with a similar, severe SRP phenotype. In a mouse mutation screen for developmental phenotypes, we identified a mutation in Wdr35 as the cause of midgestation lethality, with abnormalities characteristic of defects in the Hedgehog signaling pathway. We show that endogenous WDR35 localizes to cilia and centrosomes throughout the developing embryo and that human and mouse fibroblasts lacking the protein fail to produce cilia. Through structural modeling, we show that WDR35 has strong homology to the COPI coatamers involved in vesicular trafficking and that human SRP mutations affect key structural elements in WDR35. Our report expands, and sheds new light on, the pathogenesis of the SRP spectrum of ciliopathies.


Effects of ranolazine on torsades de pointes tachycardias in a healthy isolated rabbit heart model.
Torsades de pointes (TdP) tachycardias are triggered, polymorphic ventricular arrhythmias arising from early afterdepolarizations (EADs) and increased dispersion of repolarization. Ranolazine is a new agent which reduces pathologically elevated late INa but also IKr . Aim of this study was to evaluate the effects of ranolazine in a validated isolated Langendorff-perfused rabbit heart model. TdP was reproducibly induced with d-sotalol (10(-4) mol/L) and low potassium (K) (1.0 mmol/L for 5 min, pacing at CL 1000 ms). In 10 hearts, ECG and 8 epi- and endocardial monophasic action potentials were recorded. Action potential duration (APD) was measured at 90% repolarization and dispersion defined as APD max-min. D-sotalol prolonged APD90 and increased dispersion of APD90 , simultaneously causing EADs and induction of TdP. The combination of d-sotalol and two concentrations of ranolazine did not increase dispersion of ventricular APD90 as compared to vehicle. Ranolazine at 5 μmol/L did not cause additional induction of EADs and/or TdP but also did not significantly suppress arrhythmogenic triggers. The higher concentration of ranolazine (10 μmol/L) in combination with d-sotalol caused further prolongation of APD90 , at the same time reduction in APD90 dispersion. In parallel, the incidence of EADs was reduced and an antitorsadogenic effect was seen. In the healthy isolated rabbit heart (where late INa is not elevated), ranolazine does not cause proarrhythmia but exerts antiarrhythmic effects in a dose-dependent manner against d-sotalol/low K-induced TdP. This finding-despite additional APD prolongation-supports the safety of a combined use of both drugs and merits clinical investigation.


Biophysical characterization of recombinant human Bcl-2 and its interactions with an inhibitory ligand, antimycin A.
Apoptosis is an essential physiological process, regulated by the family of Bcl-2-related proteins. However, the molecular mechanism by which Bcl-2 regulates apoptosis still remains elusive. Here we report the functional studies of recombinant human Bcl-2 with the deletion of 22 residues at the C-terminal membrane-anchoring region (rhBcl-2Delta22). Characterization of rhBcl-2Delta22 showed that the recombinant protein is homogeneous and monodisperse in nondenaturing solutions, stable at room temperature in the presence of a metal chelator, and an alpha-helical protein with unfolding of secondary structure at a T(m) of 62.8 degrees C. Optimal membrane pore formation by rhBcl-2Delta22 required negatively charged phospholipids. The existence of a hydrophobic groove in rhBcl-2Delta22 was demonstrated by the fluorescence enhancement of the hydrophobic ANS probe with which a pro-apoptotic Bak BH3 peptide competed. The respiratory inhibitor antimycin A also bound to the hydrophobic groove of rhBcl-2Delta22 with a K(d) of 0.82 microM. The optimal binding conformation of antimycin A was predicted from molecular docking of antimycin A with the hBcl-2 model created by homology modeling. Antimycin A selectively induces apoptosis in cells overexpressing Bcl-2, suggesting that hydrophobic groove-binding compounds may act as selective apoptotic triggers in tumor cells.


Computational approach for deriving cancer progression roadmaps from static sample data.
As with any biological process, cancer development is inherently dynamic. While major efforts continue to catalog the genomic events associated with human cancer, it remains difficult to interpret and extrapolate the accumulating data to provide insights into the dynamic aspects of the disease. Here, we present a computational strategy that enables the construction of a cancer progression model using static tumor sample data. The developed approach overcame many technical limitations of existing methods. Application of the approach to breast cancer data revealed a linear, branching model with two distinct trajectories for malignant progression. The validity of the constructed model was demonstrated in 27 independent breast cancer data sets, and through visualization of the data in the context of disease progression we were able to identify a number of potentially key molecular events in the advance of breast cancer to malignancy.


CD4+CD25+ Regulatory T Cells Decreased CD8+IL-4+Cells in a Mouse Model of Allergic Asthma.
Interleukin (IL)-4-producing-CD8 (cytotoxic T cells, Tc) contribute to lung eosinophilia and airway hyper-responsiveness (AHR) to an antigen. CD4+CD25+ regulatory T cells (Tregs) attenuate airway inflammation and AHR. This study investigated whether Tregs decrease Tc2frequencies in ovalbumin (OVA)-induced asthma model of mice. Female C57BL/6 mice were sensitized with OVA intraperitoneally and challenged with OVA intranasally to induce allergic asthma model. Tregs were sorted by fluorescence activated cell sorting (FACS) and magnetic activated cell sorting (MACS) microbeads. OVA-sensitized mice were injected with Tregs or phosphate buffer saline (PBS) by tail vein ahead of the first challenge. Airway inflammation and airway hyper-responsiveness (AHR)were evaluated by histological analysis and invasive method, respectively. OVA-specific IgE and cytokine levels were detected by ELISA. Flow cytometry was used to detect the percentages of Tc1 and Tc2. Gata3 and T-bet mRNA was determined by quantitative PCR (qPCR). OVA-sensitized and challenged mice displayed typical asthma features, which included eosinophilic airway inflammation, higher levels of Th2 cytokines and AHR. Gata3 mRNA, Tc2 frequencies and OVA-specific IgE levels were significantly increased in OVA-sensitized and challenged mice. Compared to PBS treatment, Tregs decreased Tc2 frequencies, airway inflammation, Th2 cytokine levels and AHR in OVA-sensitized and challenged mice. IL-13 levels were negatively correlated with Tc1 frequencies and with IFNg levels in experimental mice. Our results demonstrated that Tregs could prevent airway inflammation and AHR by decreasing Tc2 frequencies and cytokine levels in OVA-induced asthma model of mice, supporting Tregmight be as a potent therapeutic target for alleviating airway inflammation and AHR.


A renaissance for YES in cancer.
Most of our understanding regarding the involvement of SRC-family tyrosine kinases in cancer has stemmed from studies focused on the prototypical SRC oncogene. However, emerging research has shed light on the important role of YES signaling in oncogenic transformation, tumor growth, metastatic progression, and resistance to various cancer therapies. Clinical evidence indicates that dysregulated expression or activity of YES is a frequent occurrence in human cancers and is associated with unfavorable outcomes. These findings provide a compelling rationale for specifically targeting YES in certain cancer subtypes. Here, we review the crucial role of YES in cancer and discuss the challenges associated with translating preclinical observations into effective YES-targeted therapies.


Service screening with mammography. Long-term effects on breast cancer mortality in the county of Gävleborg, Sweden.
In the current study the long-term effects of a pilot service screening programme in the Swedish county of Gävleborg were studied. Women aged 40-64 years in 13 sub-areas were followed from start of screening between 1974 and 1979. Two control groups were used for comparison; the neighbouring counties and all of Sweden. A reference period prior to the study period was used to facilitate an adjustment for possible differences in baseline breast cancer mortality. Only deaths from breast cancer diagnosed after the first invitation to screening were analysed. Two outcome measures were used for breast cancer mortality; the underlying cause of death and excess mortality. Using the neighbouring counties as a control group, the relative risk, after 22 years of follow-up, of 10 years of screening was estimated at 0.84 (95% CI 0.71-1.00) using excess mortality. Due to lead time bias the relative risk was overestimated by 4%. Hence, a significant 20% reduction of breast cancer excess mortality was found.


Molecular pathways: gene-environment interactions regulating dietary fiber induction of proliferation and apoptosis via butyrate for cancer prevention.
Gene-environment interactions are so numerous and biologically complicated that it can be challenging to understand their role in cancer. However, dietary fiber and colorectal cancer prevention may represent a tractable model system. Fiber is fermented by colonic bacteria into short-chain fatty acids such as butyrate. One molecular pathway that has emerged involves butyrate having differential effects depending on its concentration and the metabolic state of the cell. Low-moderate concentrations, which are present near the base of colonic crypts, are readily metabolized in the mitochondria to stimulate cell proliferation via energetics. Higher concentrations, which are present near the lumen, exceed the metabolic capacity of the colonocyte. Unmetabolized butyrate enters the nucleus and functions as a histone deacetylase (HDAC) inhibitor that epigenetically regulates gene expression to inhibit cell proliferation and induce apoptosis as the colonocytes exfoliate into the lumen. Butyrate may therefore play a role in normal homeostasis by promoting turnover of the colonic epithelium. Because cancerous colonocytes undergo the Warburg effect, their preferred energy source is glucose instead of butyrate. Consequently, even moderate concentrations of butyrate accumulate in cancerous colonocytes and function as HDAC inhibitors to inhibit cell proliferation and induce apoptosis. These findings implicate a bacterial metabolite with metaboloepigenetic properties in tumor suppression.


Auditory processing following infantile spasms: An event-related potential study.
To investigate acoustic auditory processing in patients with recent infantile spasms (IS). Patients (n = 22; 12 female; median age 8 months; range 5-11 months) had normal preceding development, brain magnetic resonance imaging (MRI), and neurometabolic testing (West syndrome of unknown cause, uWS). Controls were healthy babies (n = 22; 11 female; median age 6 months; range 3-12 months). Event-related potentials (ERPs) and psychometry (Bayley Scales of Infant Development, Second Edition, BSID-II) took place at a month following IS remission. Following a repeated pure tone, uWS patients showed less suppression of the N100 at the mid-temporal electrodes (p = 0.006), and a prolonged response latency (p = 0.019). Their novelty P300 amplitude over the mid-temporal electrodes was halved (p = 0.001). The peak of the novelty P300 to environmental broadband sounds emerged later over the left temporal lobe in patients (p = 0.015), the lag correlating with duration of spasms (r = 0.547, p = 0.015). BSID-II scores were lower in patients (p < 0.001), with no correlation to ERP. Complex acoustic information is processed poorly following IS. This would impair language. Treatment did not reverse this phenomenon, but may have limited its severity. The data are most consistent with altered connectivity of the cortical acoustic processing areas induced by IS.


Sleep pattern and staging in elite adolescent rugby players during the in-season competitive phase compared to an age matched non-athlete population.
Young athletes must contend with the constraints of elite sports on top of school commitments and the physiological processes associated with adolescence. This study assessed week and weekend sleep and schedule of activities in elite adolescent rugby players during the in-season competitive phase compared with age-matched non-athlete controls. 32 adolescents (GR: 16 elite rugby players, GC: 16 controls) from the same boarding school filled out a daily schedule of activities and a sleep diary, and wore a multichannel electroencephalogram for 14 days. They later filled out questionnaires on their sleep quality, sleepiness, and perceived stress. Both groups showed insufficient sleep duration during the week (<7 h). Only GC caught up on their sleep debt during the weekend (increased TIB, TST and time spent in REM sleep, all p < 0.001). Weekend TIB, TST and, N3 sleep remained similar to that for weekdays in GR. However, GR experienced lower sleep quality (decreased SE, increased WASO, all p < 0.01) and a decrease in REM sleep (p < 0.01). Schedules of activities showed an increase in time spent on overall activities during the weekend, mainly due to competition and sport-specific travel, which resulted in a decrease in sleep opportunity time compared with GC (p < 0.001). Δ sleep opportunity time (weekend-week) was associated with Δ TST (weekend-week), and Δ TST (weekend-week) was associated with sleepiness and perceived stress. Busy schedules during the competitive season decreased sleep opportunity time and prevented elite adolescent rugby players from catching up on their sleep at weekends.HIGHLIGHTSAdolescent rugby players have insufficient sleep during both week and weekend of the in-season competitive phaseCongested schedule prevented elite adolescent rugby players from catching up on their sleep at weekends.Sleep considerations are necessary when planning the weekend sporting schedules.


Should NICE reconsider the 2016 UK guidelines on TB contact tracing? A cost-effectiveness analysis of contact investigations in London.
In January 2016, clinical TB guidance in the UK changed to no longer recommend screening contacts of non-pulmonary, non-laryngeal (ETB) index cases. However, no new evidence was cited for this change, and there is evidence that screening these contacts may be worthwhile. The objective of this study was to estimate the cost-effectiveness of screening contacts of adult ETB cases and adult pulmonary or laryngeal TB (PTB) cases in London, UK. We carried out a cross-sectional analysis of data collected on TB index cases and contacts in the London TB register and an economic evaluation using a static model describing contact tracing outcomes. Incremental cost-effectiveness ratios (ICERs) were calculated using no screening as the baseline comparator. All adult TB cases (≥15 years old) in London from 2012 to 2015, and their contacts, were eligible (2465/5084 PTB and 2559/6090 ETB index cases were included). Assuming each contact with PTB infects one person/month, the ICER of screening contacts of ETB cases was £78 000/quality-adjusted life-years (QALY) (95% CI 39 000 to 140 000), and screening contacts of PTB cases was £30 000/QALY (95% CI 18 000 to 50 000). The ICER of screening contacts of ETB cases was £30 000/QALY if each contact with PTB infects 3.4 people/month. Limitations of this study include the use of self-reported symptomatic periods and lack of knowledge about onward transmission from PTB contacts. Screening contacts of ETB cases in London was almost certainly not cost-effective at any conventional willingness-to-pay threshold in England, supporting recent changes to National Institute for Health and Care Excellence national guidelines.


The Effects of Augmented Reality Visual Cues on Turning in Place in Parkinson's Disease Patients With Freezing of Gait.
Background: Turning in place is particularly bothersome for patients with Parkinson's disease (PD) experiencing freezing of gait (FOG). Cues designed to enforce goal-directed turning are not yet available. Objectives: Assess whether augmented reality (AR) visual cues improve FOG and turning in place in PD patients with FOG. Methods: Sixteen PD patients with FOG performed a series of 180° turns under an experimental condition with AR visual cues displayed through a HoloLens and two control conditions (one consisting of auditory cues and one without any cues). FOG episodes were annotated by two independent raters from video recordings. Motion data were measured with 17 inertial measurement units for calculating axial kinematics, scaling, and timing of turning. Results: AR visual cues did not reduce the percent time frozen (p = 0.73) or the number (p = 0.73) and duration (p = 0.78) of FOG episodes compared to the control condition without cues. All FOG parameters were higher with AR visual cues than with auditory cues [percent time frozen (p = 0.01), number (p = 0.02), and duration (p = 0.007) of FOG episodes]. The AR visual cues did reduce the peak angular velocity (visual vs. uncued p = 0.03; visual vs. auditory p = 0.02) and step height (visual vs. uncued p = 0.02; visual vs. auditory p = 0.007), and increased the step height coefficient of variation (visual vs. uncued p = 0.04; visual vs. auditory p = 0.01) and time to maximum head-pelvis separation (visual vs. uncued p = 0.02; visual vs. auditory p = 0.005), compared to both control conditions. Conclusions: The AR visual cues in this study did not reduce FOG, and worsened some measures of axial kinematics, and turn scaling and timing. Stimulating goal-directed turning might, by itself, be insufficient to reduce FOG and improve turning performance. Trial Registration: This study was registered in the Dutch trial registry (NTR6409; 2017-02-16).


Benzyl isothiocyanate (BITC) inhibits migration and invasion of human gastric cancer AGS cells via suppressing ERK signal pathways.
Metastasis suppressors and associated other regulators of cell motility play a critical initial role in tumor invasion and metastases. Benzyl isothiocyanate (BITC) is a hydrolysis compound of glucotropaeolin in dietary cruciferous vegetables. BITC has been found to exhibit prevention of cancers in laboratory animals and might also be chemoprotective in humans. Here, the purpose of this study was to investigate the effects of BITC on cell proliferation, migration, invasion and mitogen-activated protein kinase (MAPK) pathways of AGS human gastric cancer cells. Wound healing and Boyden chamber (migration and invasion) assays demonstrated that BITC exhibited an inhibitory effect on the abilities of migration and invasion in AGS cancer cells. BITC suppressed cell migration and invasion of AGS cells in a dose-dependent manner. Results from Western blotting indicated that BITC exerted an inhibitory effect on the ERK1/2, Ras, GRB2, Rho A, iNOS, COX-2 for causing the inhibitions of MMP-2, -7 and -9 then followed by the inhibitions of invasion and migration of AGS cells in vitro. BITC also promoted MKK7, MEKK3, c-jun, JNK1/2, VEGF, Sos1, phosphoinositide 3-kinase (PI3K), PKC, nuclear factor-kappaB (NF-κB) p65 in AGS cells. Results from real-time polymerized chain reaction (PCR) showed that BITC inhibited the gene expressions of MMP-2,-7 -9, FAK, ROCK1 and RhoA after BITC treatment for 24 and 48 hours in AGS cells. Taken together, the finding may provide new mechanisms and functions of BITC, which inhibit migration and invasion of human gastric cancer AGS cells.


Cocaine-mediated induction of platelet-derived growth factor: implication for increased vascular permeability.
Neuroinflammation associated with advanced HIV-1 infection is often exacerbated in cocaine-abusing, HIV-infected patients. The underlying mechanisms could, in part, be attributed to the increased impairment of blood brain barrier integrity in the presence of cocaine. Platelet-derived growth factor (PDGF) has been implicated in several pathologic conditions, specifically attributable to its potent mitogenic effects. Its modulation by drug abuse, however, has received very little attention. In the present study, we demonstrated cocaine-mediated induction of PDGF-BB in human brain microvascular endothelial cells through the binding to its cognate σ receptor. Furthermore, this effect was mediated, with subsequent activation of mitogen-activated protein kinases and Egr-1 pathways, culminating ultimately into increased expression of PDGF-BB. Cocaine exposure resulted in increased permeability of the endothelial barrier, and this effect was abrogated in mice exposed to PDGF-BB neutralizing antibody, thus underscoring its role as a vascular permeant. In vivo relevance of these findings was further corroborated in cocaine-treated mice that were administered neutralizing antibody specific for PDGF-BB as well as in Egr-1(-/-) mice. Understanding the regulation of PDGF-BB expression may provide insights into the development of potential therapeutic targets for neuroinflammation associated with HIV infection and drug abuse.


Effector role of neonatal hepatic CD8+ lymphocytes in epithelial injury and autoimmunity in experimental biliary atresia.
Lymphocytes populate the livers of infants with biliary atresia, but it is unknown whether neonatal lymphocytes regulate pathogenesis of disease. Here, we investigate this question by examining the role of T lymphocytes in the destruction of extrahepatic bile ducts of neonatal mice using an experimental model of biliary atresia. Inoculation of neonatal mice with rhesus rotavirus followed by multistaining flow cytometry to quantify expression of interferon-gamma by hepatic lymphocytes, and real-time polymerase chain reaction for mRNA expression of pro-inflammatory cytokines. This was followed by determining the consequences of antibody-mediated depletion of lymphocyte subtypes on the development of biliary obstruction, and coculture and cell transfer experiments to investigate the effector role of lymphocyte subtypes on neonatal biliary disease. Rotavirus infection results in overexpression of interferon-gamma by neonatal hepatic T cells. Among these cells, depletion of CD4(+) cells did not change the course of inflammatory injury and obstruction of neonatal bile ducts. In contrast, loss of CD8(+) cells remarkably suppressed duct injury, prevented luminal obstruction, and restored bile flow. Coculture experiments showed that rotavirus-primed, but not naïve, CD8(+) cells were cytotoxic to cholangiocytes. In adoptive transfer experiments, we found that primed CD8(+) cells preferentially homed to extrahepatic bile ducts of neonatal mice and invaded their epithelial lining. Primed neonatal CD8(+) cells can activate a pro-inflammatory program, target diseased and healthy duct epithelium, and drive the phenotypic expression of biliary atresia, thus constituting a potential therapeutic target to halt disease progression.


Toward more definitive clinical trials.
The randomized clinical trial is an essential evaluation tool in the health care field. Unfortunately, the majority of trials performed have too few patients to reach a conclusion regarding the treatments under study. In many instances, the multicenter trial represents the only way to obtain the necessary numbers. Such trials are difficult to organize and costly to carry out, but they have strengths not found in the ordinary single center trial. The multiplicity of disciplines required usually leads to better design and more careful execution than can be achieved with the resources of a single center. The heterogeneity of the population and multiplicity of clinics applying the study treatments provide a more realistic test of the therapies in question than is possible in a single clinic. Sponsors of trials need to stimulate more research into the methodology of multicenter trials, including investigation of the methods of organizing and carrying out such trials in a cost-effective manner. In addition, a communications network should be developed to promote the field of clinical trials as a discipline and to facilitate the identification and dissemination of information on their design and conduct.


Menstrual blood-derived mesenchymal stromal cells: impact of preconditioning on the cargo of extracellular vesicles as potential therapeutics.
Mesenchymal stromal cells (MSCs) have been shown to exert their therapeutic effects through the secretion of broad spectrum of paracrine factors, including extracellular vesicles (EVs). Accordingly, EVs are being pursued as a promising alternative to cell-based therapies. Menstrual blood-derived stromal cells (MenSCs) are a type of MSC that, due to their immunomodulatory and regenerative properties, have emerged as an innovative source. Additionally, new strategies of cell priming may potentially alter the concentration and cargo of released EVs, leading to modification of their biological properties. In this study, we aimed to characterize the EVs released by MenSCs and compare their therapeutic potential under three different preconditioning conditions (proinflammatory stimuli, physioxia, and acute hypoxia). MenSCs were isolated from five healthy women. Following culturing to 80% confluence, MenSCs were exposed to different priming conditions: basal (21% O2), proinflammatory stimuli (IFNγ and TNFα, 21% O2), physioxia (1-2% O2), and acute hypoxia (< 1% O2) for 48-72 h. Conditioned media from MenSCs was collected after 48 h and EVs were isolated by a combination of ultra-filtration and differential centrifugation. An extensive characterization ranging from nano-flow cytometry (nFC) to quantitative high-throughput shotgun proteomics was performed. Bioinformatics analyses were used to derive hypotheses on their biological properties. No differences in the morphology, size, or number of EVs released were detected between priming conditions. The proteome analysis associated with basal MenSC-EVs prominently revealed their immunomodulatory and regenerative capabilities. Furthermore, quantitative proteomic analysis of differentially produced MenSC-EVs provided sufficient evidence for the utility of the differential preconditioning in purpose-tailoring EVs for their therapeutic application: proinflammatory priming enhanced the anti-inflammatory, regenerative and immunomodulatory capacity in the innate response of EVs, physioxia priming also improves tissue regeneration, angiogenesis and their immunomodulatory capacity targeting on the adaptive response, while acute hypoxia priming, increased hemostasis and apoptotic processes regulation in MenSC-EVs, also by stimulating immunomodulation mainly through the adaptive response. Priming of MenSCs under proinflammatory and hypoxic conditions affected the cargo proteome of EVs released, resulting in different therapeutic potential, and thus warrants experimental exploration with the aim to generate better-defined MSC-derived bioproducts.


Cytomorphologic features in thyroid nodules read as "suspicious for malignancy" on cytology may predict thyroid cancers with the BRAF mutation.
Some morphologic parameters have been studied to help predict the BRAF(V600E) mutation using cytopathologic specimens, which can indicate which nodules should undergo further testing. The aim of this study was to investigate the value of cytomorphologic parameters to predict the BRAF(V600E) mutation in nodules read as "suspicious for malignancy" on cytology. This study included 142 resected nodules which were diagnosed as "suspicious for malignancy" on cytology in 142 patients. At our institution, BRAF(V600E) mutation analysis was performed at the request of the referring clinicians based on the clinical features of the patients, or the judgment of the radiologists performing US-FNA because suspicious US features were observed on the targeted nodule during this study period. Cytology smears were re-reviewed to assess the presence and amount of polygonal eosinophilic (plump) cells and microfollicles, and the presence of intranuclear pseudoinclusions, irregular nuclear membranes, nuclear grooves, sickles cells, psammoma bodies, and cystic changes. We evaluated the diagnostic performances of the cytomorphologic features to predict the BRAF(V600E) mutation. Polygonal eosinophilic (plump) cells, microfollicles, intranuclear pseudoinclusions, sickle cells, and cystic changes were significantly associated with the BRAF(V600E) mutation. The mutation was not present in all 6 thyroid nodules with microfollicles larger than 20% on cytology. Additionally, polygonal eosinophilic (plump) cells larger than 20%, cystic changes, and sickle cells on cytology had a high specificity of 95%, 96.7%, and 81.7%, respectively. Excluding 6 nodules with microfollicles larger than 20% on cytology, there were 82 (60.3%) nodules with the BRAF(V600E) mutation among the 136 nodules. Among the 136 nodules, there were 95 nodules with polygonal eosinophilic (plump) cells larger than 20%, cystic changes, or sickle cells on cytology. Of the 95 nodules, 69 (72.6%) had the mutation. Cytomorphologic features can help select nodules for the BRAF(V600E) mutation test among nodules read as "suspicious for malignancy" on cytology.