Tabalumab in rheumatoid arthritis patients with an inadequate response to methotrexate and naive to biologic therapy: a phase II, randomized, placebo-controlled trial.
Tabalumab, a fully human IgG4 monoclonal antibody, neutralizes soluble and membrane-bound BAFF. The aim of this study was to examine the tolerability and efficacy of tabalumab in patients with active rheumatoid arthritis receiving methotrexate. In this randomized, double-blind, placebo-controlled, parallel, multiple-dose study, patients who were naive to biologic therapy received infusions of tabalumab (30, 60, or 160 mg) or placebo at weeks 0, 3, and 6 in combination with methotrexate and were evaluated for 24 weeks. The primary efficacy end point was the percentage of patients meeting American College of Rheumatology 20% improvement criteria (achieving an ACR20 response) at week 16. At week 16, the percentages of patients achieving an ACR20 response in the 30-mg (57.6%), 60-mg (67.6%), and 160-mg (51.5%) groups were significantly greater than the percentage of patients achieving an ACR20 response in the placebo group (29.4%; P<0.05). There were initial transient increases from baseline in the frequency of CD20+ and IgD+/CD27- B cells, followed by reductions, although B cells were not completely depleted. Also, the frequency of IgD-/CD27+ B cells increased in all tabalumab groups compared with the placebo group and returned toward baseline levels by the end of the study. The incidence of adverse events was similar across all treatment groups; no deaths occurred. Serum IgM levels decreased significantly in all tabalumab groups combined compared with the placebo group. There were no significant decreases in serum IgG or IgA levels in the tabalumab groups compared with the placebo group. Tabalumab treatment significantly reduces the signs and symptoms of rheumatoid arthritis and has a safety profile similar to that seen with placebo treatment.


The emotional costs of caring incurred by men and women in the British labour market.
This study investigates whether men and women in caring occupations experience more negative job-related feelings at the end of the day compared to the rest of the working population. The data are from Wave Nine of the British Household Panel Survey (1999) where respondents were asked whether, at the end of the working day, they tended to keep worrying or have trouble unwinding, and the extent to which work left them feeling exhausted or "used up." Their responses to these questions were used to develop ordinal dependent variables. Control variables in the models include: number of children, age, hours worked per week, managerial responsibilities and job satisfaction, all of which have been shown in previous research to be significantly related to "job burnout." The results are that those in caring occupations are more likely to feel worried, tense, drained and exhausted at the end of the working day. Women in particular appear to pay a high emotional cost for working in caring occupations. Men do not emerge unscathed, but report significantly lower levels of worry and exhaustion at the end of the day than do women.


Role of the bacteriophage T7 and T4 single-stranded DNA-binding proteins in the formation of joint molecules and DNA helicase-catalyzed polar branch migration.
Bacteriophage T7 gene 2.5 single-stranded DNA-binding protein and gene 4 DNA helicase together promote pairing of two homologous DNA molecules and subsequent polar branch migration (Kong, D., and Richardson, C. C. (1996) EMBO J. 15, 2010-2019). In this report, we show that gene 2.5 protein is not required for the initiation or propagation of strand transfer once a joint molecule has been formed between the two DNA partners, a reaction that is mediated by the gene 2.5 protein alone. A mutant gene 2.5 protein, gene 2.5-Delta21C protein, lacking 21 amino acid residues at its C terminus, cannot physically interact with gene 4 protein. Although it does bind to single-stranded DNA and promote the formation of joint molecule via homologous base pairing, subsequent strand transfer by gene 4 helicase is inhibited by the presence of the gene 2.5-Delta21C protein. Bacteriophage T4 gene 32 protein likewise inhibits T7 gene 4 protein-mediated strand transfer, whereas Escherichia coli single-stranded DNA-binding protein does not. The 63-kDa gene 4 protein of phage T7 is also a DNA primase in that it catalyzes the synthesis of oligonucleotides at specific sequences during translocation on single-stranded DNA. We find that neither the rate nor extent of strand transfer is significantly affected by concurrent primer synthesis. The bacteriophage T4 gene 41 helicase has been shown to catalyze polar branch migration after the T4 gene 59 helicase assembly protein loads the helicase onto joint molecules formed by the T4 UvsX and gene 32 proteins (Salinas, F., and Kodadek, T. (1995) Cell 82, 111-119). We find that gene 32 protein alone forms joint molecules between partially single-stranded homologous DNA partners and that subsequent branch migration requires this single-stranded DNA-binding protein in addition to the gene 41 helicase and the gene 59 helicase assembly protein. Similar to the strand transfer reaction, strand displacement DNA synthesis catalyzed by T4 DNA polymerase also requires the presence of gene 32 protein in addition to the gene 41 and 59 proteins.


Relationship of delayed parenteral nutrition protocol with the clinical outcomes in a medical intensive care unit.
Although parenteral nutrition (PN) is an important treatment for patients who are unable to tolerate enteral nutrition (EN), recent international guidelines recommended that PN should be reserved and initiated only after 7 days in well-nourished patients. This retrospective study was conducted to analyze the effect on clinical outcomes of a PN protocol with PN starting 4 days after admission to the intensive care unit (ICU). Eighty-one patients who were admitted from January to March of 2012 were included in the pre-protocol group, and 74 patients who were admitted from April to June of 2012 were included in the post-protocol group. There were no significant differences between the two groups when the age, gender, and admission Acute Physiology and Chronic Health Evaluation (APACHE) II scores were compared. Significantly fewer patients in the post-protocol group were provided PN (58.1% vs. 81.3%, p = 0.002), which was initiated significantly later than in the pre-protocol group (2.7 ± 2.2 days vs. 1.9 ± 2.0 days, p = 0.046). Five patients (6.2%) in the pre-protocol group acquired central line-associated bloodstream infection (CLA-BSI) in the ICU, yet none of the patients in the post-protocol group developed CLA-BSI (p = 0.03). The duration of antibiotic therapy and ICU stay were significantly shorter in the post-protocol group than in the pre-protocol group. By delaying initiation of PN, short-term clinical outcomes including incidence of CLA-BSI, antibiotic use, and ICU stay might be improved. Further research should be conducted to investigate the long-term effects of the decreased nutrient intake resulting from delayed PN.


Visualization of Vascular and Parenchymal Regeneration after 70% Partial Hepatectomy in Normal Mice.
A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the silicone compound, via a 26 G catheter, into the portal or hepatic vein. After silicone injection, organs were explanted and prepared for ex-vivo micro-CT (µCT) scanning. The silicone injection procedure is technically challenging. Achieving a successful outcome requires extensive microsurgical experience from the surgeon. One of the challenges of this procedure involves determining the adequate perfusion rate for the silicone compound. The perfusion rate for the silicone compound needs to be defined based on the hemodynamic of the vascular system of interest. Inappropriate perfusion rate can lead to an incomplete perfusion, artificial dilation and rupturing of vascular trees. The 3D reconstruction of the vascular system was based on CT scans and was achieved using preclinical software such as HepaVision. The quality of the reconstructed vascular tree was directly related to the quality of silicone perfusion. Subsequently computed vascular parameters indicative of vascular growth, such as total vascular volume, were calculated based on the vascular reconstructions. Contrasting the vascular tree with silicone allowed for subsequent histological work-up of the specimen after µCT scanning. The specimen can be subjected to serial sectioning, histological analysis and whole slide scanning, and thereafter to 3D reconstruction of the vascular trees based on histological images. This is the prerequisite for the detection of molecular events and their distribution with respect to the vascular tree. This modified silicone injection procedure can also be used to visualize and reconstruct the vascular systems of other organs. This technique has the potential to be extensively applied to studies concerning vascular anatomy and growth in various animal and disease models.


The implementation of a battery of in vivo and in vitro bioassays to assess river water for estrogenic endocrine disrupting chemicals.
Previous research has shown that accurate evaluation of environmental water samples for estrogenic activity requires a panel of in vitro and in vivo bioassays, which are based on different molecular and cellular action mechanisms. In the current study, a test battery containing four assays was used to analyze water from the Eerste River, South Africa for estrogenicity. Three sites were used for analysis, namely Jonkershoek (control site situated in the mountains at the origin of the Eerste River), sewage effluent from Stellenbosch sewage treatment works and Spier site (sampling site on the Eerste River downstream from Stellenbosch). Estrogenicity was determined using an estrone enzyme linked immuno sorbent assay (ELISA), estrogen induced proliferation of human breast cancer adenocarcinoma cells (MCF-7) also known as the E-SCREEN, estrogen induced suppression of estrogen receptor alpha protein expression (ER-α) in MCF-7 cells (ERα assay) and by monitoring estrogen induced vitellogenin (VTG) synthesis in juvenile Oreochromis mossambicus (VTG assay). Low concentrations of estrone (ranging between 1.4 and 2.2 ng/l) near the detection limit of the assay were detected in samples collected from Jonkershoek. Water from this site shows no estrogenicity in the E-SCREEN, ERα assay or VTG synthesis bioassay. The estrone concentrations in the sewage effluent extracts, as well as Spier site extracts, ranged between 14.7 and 19.4 ng/l. The assays using ERα induction by the MCF-7 cell line, MCF-7 proliferation and in vivo VTG synthesis by juvenile tilapia showed that these samples are estrogenic. The results obtained for the assays in the battery are comparable.


Pathophysiology of thrombotic thrombocytopenic purpura.
Thrombotic thrombocytopenic purpura (TTP) is a disorder with characteristic von Willebrand factor (VWF)-rich microthrombi affecting the arterioles and capillaries of multiple organs. The disorder frequently leads to early death unless the patients are treated with plasma exchange or infusion. Studies in the last decade have provided ample evidence to support that TTP is caused by deficiency of a plasma metalloprotease, ADAMTS13. When exposed to high shear stress in the microcirculation, VWF and platelets are prone to form aggregates. This propensity of VWF and platelet to form microvascular thrombosis is mitigated by ADAMTS13, which cleaves VWF before it is activated by shear stress to cause platelet aggregation in the circulation. Deficiency of ADAMTS13, due to autoimmune inhibitors in patients with acquired TTP and mutations of the ADAMTS13 gene in hereditary cases, leads to VWF-platelet aggregation and microvascular thrombosis of TTP. In this review, we discuss the current knowledge on the pathogenesis, diagnosis and management of TTP, address the ongoing controversies, and indicate the directions of future investigations.


Climate-forced variability of ocean hypoxia.
Oxygen (O(2)) is a critical constraint on marine ecosystems. As oceanic O(2) falls to hypoxic concentrations, habitability for aerobic organisms decreases rapidly. We show that the spatial extent of hypoxia is highly sensitive to small changes in the ocean's O(2) content, with maximum responses at suboxic concentrations where anaerobic metabolisms predominate. In model-based reconstructions of historical oxygen changes, the world's largest suboxic zone, in the Pacific Ocean, varies in size by a factor of 2. This is attributable to climate-driven changes in the depth of the tropical and subtropical thermocline that have multiplicative effects on respiration rates in low-O(2) water. The same mechanism yields even larger fluctuations in the rate of nitrogen removal by denitrification, creating a link between decadal climate oscillations and the nutrient limitation of marine photosynthesis.


Turning the Tables: Power Relations Between Consumer Researchers and Other Mental Health Researchers.
A crucial development resulting from consumer involvement in mental health services has been engagement as active participants in mental health research, often conducted in collaboration with mental health researchers representing the health disciplines (referred to in this paper as 'other' researchers). Despite progress in mental health consumer research, unequal power relations continue to pose a major barrier. Although power issues are discussed in the literature, there is little research from the perspective of other mental health researchers who have collaborated with consumers on research projects. This qualitative study explored other mental health researchers' perspectives on the role of power in collaborative research with consumers. Semi-structured interviews were completed with 11 other mental health researchers. Thematic analysis of the transcript version of interview recordings was conducted. The findings were grounded in 'the table' as a literal and metaphorical site of power relations. The umbrella theme was prominence and presence (of consumers) at the table, followed by subthemes on barriers (tokenism, undermined potential) and surmounting them through reworking power (critical mass and openness to power dynamics). Overall it was found that while there continue to be significant power-related barriers to further building of robust collaborative research with consumers in mental health, there are several avenues that should be considered, much more assertively, to disrupt and transcend them.


The NMR structure of the 47-kDa dimeric enzyme 3,4-dihydroxy-2-butanone-4-phosphate synthase and ligand binding studies reveal the location of the active site.
Recent developments in NMR have extended the size range of proteins amenable to structural and functional characterization to include many larger proteins involved in important cellular processes. By applying a combination of residue-specific isotope labeling and protein deuteration strategies tailored to yield specific information, we were able to determine the solution structure and study structure-activity relationships of 3,4-dihydroxy-2-butanone-4-phosphate synthase, a 47-kDa enzyme from the Escherichia coli riboflavin biosynthesis pathway and an attractive target for novel antibiotics. Our investigations of the enzyme's ligand binding by NMR and site-directed mutagenesis yields a conclusive picture of the location and identity of residues directly involved in substrate binding and catalysis. Our studies illustrate the power of state-of-the-art NMR techniques for the structural characterization and investigation of ligand binding in protein complexes approaching the 50-kDa range in solution.


Health Care Spending: Changes in the Perceptions of the Australian Public.
Increasing demand for services and rising health care costs create pressures within the Australian health care system and result in higher health insurance premiums and out-of-pocket costs for consumers. To measure changes in consumer views on the quality of the Australian health care system, contributors to rising costs and attitudes towards managing these costs. Two computer-assisted telephone interviews were conducted in 2006 (533 respondents) and 2015 (1318 respondents) and results compared. More respondents in 2015 rated the Australian health care system 'very adequate' than in 2006 (22.3% vs 8.3%; Odds Ratio OR 3.2, 99% CI 2.1, 5.1) with fewer 'concerned' or 'fairly concerned' about the health care costs (69.0% vs 85.7%; OR 0.37, 99% CI 0.25, 0.53). The 2015 respondents were more likely to identify new treatments for cancer (77% vs 65.7%; OR 1.75, 99% CI 1.30, 2.35) and community expectations for access to the latest technologies (73.8% vs 67%; OR 1.39, 99% CI 1.04, 1.86) as contributors to rising health care costs. While more 2015 respondents agreed that patients should pay a greater part of the health care costs, this remained a minority view (37.9% vs 31.7%; OR 1.32, 99% CI 0.99, 1.76). They were less likely to agree that doctors should offer medical treatments regardless of the cost and chance of benefit (63.6% vs 82.9%; OR 0.36, 99% CI 0.25, 0.50). Satisfaction with the Australian health care system has increased over time. Consumers recognise the cost pressures and have lower expectations that all services should be provided regardless of their costs and potential benefit. Public consultation on the allocation of health care resources and involvement in health care decision-making remains important. There should be community consultation about the principles and values that should guide resource allocation decisions.


Prognostic Value of Risk Factors, Calcium Score, Coronary CTA, Myocardial Perfusion Imaging, and Invasive Coronary Angiography in Kidney Transplantation Candidates.
This study sought to perform a prospective head-to-head comparison of the predictive value of clinical risk factors and a variety of cardiac imaging modalities including coronary artery calcium score (CACS), coronary computed tomography angiography (CTA), single-photon emission computed tomography (SPECT), and invasive coronary angiography (ICA) on major adverse cardiac events (MACE) and all-cause mortality in kidney transplantation candidates. Current guidelines recommend screening for coronary artery disease in kidney transplantation candidates. Furthermore, noninvasive stress imaging is recommended in current guidelines, despite its low diagnostic accuracy and uncertain prognostic value. The study prospectively evaluated 154 patients referred for kidney transplantation. All patients underwent CACS, coronary CTA, SPECT, and ICA testing. The clinical endpoints were extracted from patients' interviews, patients' records, and registries. The mean follow-up time was 3.7 years. In total, 27 (17.5%) patients experienced MACE, and 31 (20.1%) patients died during follow-up. In a time-to-event analysis, both risk factors and CACS significantly predicted death, but only CACS predicted MACE. Combining risk factors with CACS identified a very-low-risk cohort with a MACE event rate of 2.1%, and a 1.0% mortality rate per year. Of the diagnostic modalities, coronary CTA and ICA significantly predicted MACE, but only coronary CTA predicted death. In contrast, SPECT predicted neither MACE nor death. Compared with traditional risk factors and other cardiac imaging modalities, CACS and coronary CTA seem superior for risk stratification in kidney transplant candidates. Applying a combination of risk factors and CACS and subsequently coronary CTA seems to be the most appropriate strategy. (Angiographic CT of Renal Transplantation Candidate Study [ACToR]; NCT01344434).


In vivo reference point indentation reveals positive effects of raloxifene on mechanical properties following 6 months of treatment in skeletally mature beagle dogs.
Raloxifene treatment has been shown previously to positively affect bone mechanical properties following 1 year of treatment in skeletally mature dogs. Reference point indentation (RPI) can be used for in vivo assessment of mechanical properties and has been shown to produce values that are highly correlated with properties derived from traditional mechanical testing. The goal of this study was to use RPI to determine if raloxifene-induced alterations in mechanical properties occurred after 6 months of treatment. Twelve skeletally mature female beagle dogs were treated for 6 months with oral doses of saline vehicle (VEH, 1 ml/kg/day) or a clinically relevant dose of raloxifene (RAL, 0.5 mg/kg/day). At 6 months, all animals underwent in vivo RPI (10N force, 10 cycles) of the anterior tibial midshaft. RPI data were analyzed using a custom MATLAB program, designed to provide cycle-by-cycle data from the RPI test and validated against the manufacturer-provided software. Indentation distance increase (IDI), a parameter that is inversely related to bone toughness, was significantly lower in RAL-treated animals compared to VEH (-16.5%), suggesting increased bone toughness. Energy absorption within the first cycle was significantly lower with RAL compared to VEH (-21%). These data build on previous work that has documented positive effects of raloxifene on material properties by showing that these changes exist after 6 months.


Ileo-ileal intussusception in a premature neonate: an unusual cause of NEC in premature babies.
Intussusception is a rare entity in neonates. It may present with non-specific signs including abdominal distension, feeding intolerance, vomiting and bloody stools. Symptomatology is similar to Necrotizing Entero-Colitis (NEC). Ultrasound can help to establish early diagnosis in neonate. A 27-week preterm newborn was initially suspected as NEC based on abdominal distention, bilious vomiting, worsening clinical condition and dilated loops of bowel on X-ray, which turned out to be ileo-ileal intussusception. Diagnosis was made by ultrasound obtained for a palpable mass to rule out intra abdominal abscess and lack of improvement in clinical condition despite 5 days of conservative treatment. Surgery was performed consisting of removal of the necrotic intussusception area and end-to-end anastomosis and patient was discharged from hospital on day 60 of life. As a conclusion, pathological abdominal findings in preterm newborns can also be due to conditions other than NEC and ultrasound may be a useful tool for timely and accurate diagnosis.


Lysosome Membrane Permeabilization and Disruption of the Molecular Target of Rapamycin (mTOR)-Lysosome Interaction Are Associated with the Inhibition of Lung Cancer Cell Proliferation by a Chloroquinoline Analog.
Lysosomes degrade cellular proteins and organelles and regulate cell signaling by providing a surface for the formation of critical protein complexes, notably molecular target of rapamycin (mTOR) complex 1 (mTORC1). Striking differences in the lysosomes of cancer versus normal cells suggest that they could be targets for drug development. Although the lysomotropic drugs chloroquine (CQ) and hydroxychloroquine (HCQ) have been widely investigated, studies have focused on their ability to inhibit autophagy. We synthesized a novel compound, called EAD1, which is structurally related to CQ but is a 14-fold more potent inhibitor of cell proliferation. Here we find that EAD1 causes rapid relocation, membrane permeabilization (LMP), and deacidification of lysosomes, and it induces apoptosis and irreversibly blocks proliferation of human lung cancer H460, H520, H1299, HCC827, and H1703 cells. EAD1 causes dissociation of mTOR from lysosomes and increases mTOR's perinuclear versus cytoplasmic localization, changes previously shown to inactivate mTORC1. The effect on mTOR was not seen with HCQ, even at >10-fold greater concentrations. Phosphorylation of a downstream target of mTORC1, ribosomal protein S6, was inhibited by EAD1. Although EAD1 also inhibited autophagy, it retained full antiproliferative activity in autophagy-deficient H1650 lung cancer cells, which have a biallelic deletion of Atg7, and in H460 Atg7-knockout cells. As Atg7 is critical for the canonical autophagy pathway, it is likely that inhibition of autophagy is not how EAD1 inhibits cell proliferation. Further studies are needed to determine the relationship of LMP to mTORC1 disruption and their relative contributions to drug-induced cell death. These studies support the lysosome as an underexplored target for new drug development.


Results of a prospective randomized study comparing a novel retractor with a Caspar retractor in anterior cervical surgery.
Retraction injury might explain the soft tissue complications seen after anterior cervical surgery. A novel retractor system (Seex retractor system [SRS]) that uses a principle of bone fixation with rotation has been shown to reduce retraction pressure in a cadaveric model of anterior cervical decompression and fusion. To compare the conventional Cloward-style retractor (CRS) with the SRS in a prospective randomized clinical trial. After ethics and study registration (ACTRN 12608000430336), eligible patients were randomized to either the CRS or SRS before 1- or 2-level anterior cervical decompression and fusion. The pressure beneath the medial retractor blade was recorded with a thin pressure transducer strip. Postoperative sore throat, dysphagia, and dysphonia were assessed after 1, 7, and 28 days. Twenty-six patients were randomized. There were no serious complications. Complication rates were low with a trend favoring SRS that was not statistically different. Average retraction pressure with SRS was 1.9 mm Hg and with CRS was 5.6 mm Hg (P < .001 on F test; P = .002 on 2-tailed t test). Mean average peak retraction pressure with the SRS was 3.4 mm Hg and with the CRS was 20 mm Hg (P < .001 on F test; P = .005 on 2-tailed t test). The new retractor is safe, and statistically similar complication rates were observed with the 2 systems. The SRS generated significantly less retraction pressure compared with the CRS. This difference can be explained by the different principles governing the function of these retractors. Bone fixation gives stability and rotation reduces tissue pressure, both desirable in a retractor.


Failure of itraconazole to prevent T-helper type 2 cell immune deviation: Implications for chronic rhinosinusitis.
T-helper (Th) type 2 cell inflammation is the hallmark of several disease processes, including asthma, atopic dermatitis, and some forms of chronic rhinosinusitis. Itraconazole has been used as both an antifungal and an anti-inflammatory agent, with some success in many of these diseases, in part, by altering Th2 cytokine expression by T cells. It is not known whether this merely reflects inhibition of established Th2-like cells or the inhibition of differentiation of naive T cells into Th2-like cells. To evaluate the role of itraconazole in the differentiation of naive T cells during activation. Naive CD45RA+ T cells were isolated from peripheral blood mononuclear cells from healthy volunteers. Th1 and Th2 type cells were differentiated in the presence of varying concentrations of itraconazole. After stimulation with anti-CD3 and anti-CD28 beads, carboxyfluorescein succinimidyl ester dilution was performed to evaluate proliferation and intracellular cytokine staining for interleukin (IL) 4 and interferon (IFN) gamma within proliferating T cells was measured along with enzyme-linked immunosorbent assay for secreted IL-5, IL-13, and IFN gamma. Itraconazole had no effect on proliferation of unbiased, Th1, or Th2 cells. Similarly, there was no effect of itraconazole on either intracellular cytokine staining of IL-4 and IFN gamma or secreted cytokine expression of IFN gamma, IL-5, and IL-13 in any of the cell populations. Itraconazole did not alter the ability of naive T cells to proliferate or secrete cytokines under Th1 or Th2 deviating conditions in vitro. As such, reported inhibition of Th2-like lymphocyte function by itraconazole reflected action on mature effector cells and may have underscored why antifungal treatment failed in many clinical trials of eosinophilic chronic rhinosinusitis.


Human medullary thyroid carcinoma in culture provides a model relating growth dynamics, endocrine cell differentiation, and tumor progression.
We used an unique model, human medullary thyroid carcinoma (MTC) in culture (the TT line), to study features of neuroendocrine-related biochemistry in relationship to growth, differentiation, and tumor progression. Tumor tissues from patients with virulent MTC contain a very heterogeneous distribution of cells staining for calcitonin (CT) and have a high ratio of intracellular L-dopa decarboxylase activity (DDC) to CT. We found, in a culture line of MTC derived from a patient with virulent disease, that the degree of the inverse relationship between DDC and CT and the heterogeneous cellular distribution of CT probably relate to the rate of cellular growth and the biochemical set of individual cell clones. During exponential growth of the parent TT cell line, intracellular DDC and CT varied. DDC increased by 70% and CT decreased by 40%. Single time-point measurements in 54 cell clones or highly enriched cell populations revealed a more dramatic variability for CT (15-fold) than for DDC (5-fold). During growth of the clones having the highest and lowest CT measurements, respectively, inverse dynamics between DDC and CT were again found. However, each clone maintained a distinct range of CT during the entire growth curve, with a 2- to 4-fold difference in CT between the two clones throughout. In the low producing CT clone, ratios between DDC and CT rose to greater than 1.0 during growth, a very high value found before this study only in MTC tissues from patients with virulent disease. Immunohistochemical staining for CT of parent cells and clones grown on embryonic chick skin revealed increased cellular heterogeneity for CT distribution during growth. The TT line provides a powerful tool to study neuroendocrine related biochemical events in relationship to growth, differentiation, and tumor progression in MTC. Our in vitro findings in the TT line well explain observations made previously in patients. We conclude that: (1) DDC, a neural property of MTC, is an early differentiation marker as compared to CT and that the differentiation status of MTC cells varies inversely with cell growth rate; and (2) in patients with MTC, the virulence of the tumor probably varies inversely with differentiation status. The inverse ratio of DDC to CT is probably determined in MTC by the proportion of rapidly growing cells and numbers of cell clones which have a poor ability for maturation.


Assessing factors for loss to follow-up of HIV infected patients in Guinea-Bissau.
The objective of this study was to ascertain vital status of patients considered lost to follow-up at an HIV clinic in Guinea-Bissau, and describe reasons for loss to follow-up (LTFU). This study was a cross-sectional sample of a prospective cohort, carried out between May 15, 2013, and January 31, 2014. Patients lost to follow-up, who lived within the area of the Bandim Health Project, a demographic surveillance site (DSS), were eligible for inclusion. Active follow-up was attempted by telephone and tracing by a field assistant. Semi-structured interviews were done face to face or by phone by a field assistant and patients were asked why they had not shown up for the scheduled appointment. Patients were included by date of HIV testing and risk factors for LTFU were assessed using Cox proportional hazard model. Among 561 patients (69.5 % HIV-1, 18.0 % HIV-2 and 12.6 % HIV-1/2) living within the DSS, 292 patients had been lost to follow-up and were, therefore, eligible for active follow-up. Vital status was ascertained in 65.9 % of eligible patients and 42.7 % were alive, while 23.2 % had died. Information on reasons for LTFU existed for 103 patients. Major reasons were moving (29.1 %), travelling (17.5 %), and transferring to other clinics (11.7 %). A large proportion of the patients at the clinic were lost to follow-up. The main reason for this was found to be the geographic mobility of the population in Guinea-Bissau.


Rho-kinase inhibition and electromechanical coupling in rat and guinea-pig ureter smooth muscle: Ca2+-dependent and -independent mechanisms.
Recent data have shown Ca(2+)-dependent activation of Rho-kinase by sustained depolarization of arterial smooth muscle. Visceral smooth muscles, however, contract phasically in response to action potentials and it is unclear whether Ca(2+)-dependent or -independent Rho-kinase activation occurs. We have therefore investigated this, under physiologically relevant conditions, in intact ureter. Action potentials, ionic currents, Ca(2+) transients, myosin light chain (MLC) phosphorylation and phasic contraction evoked by action potentials in guinea-pig and rat ureter were investigated. In rat, but not guinea-pig ureter, three Rho-kinase inhibitors, Y-27632, HA-1077 and H-1152, significantly decreased phasic contractions and Ca(2+) transients. Voltage- and current-clamp data showed that Rho-kinase inhibition reduced the plateau component of the action potential, inhibited Ca(2+)-channels and, indirectly, Ca(2+)-activated Cl(-) channels. The Ca(2+) channel agonist Bay K8644 could reverse these effects. The K(+) channel blocker TEA could also reverse the inhibitory effect of Y-27632 on the action potential and Ca(2+) transient. Ca(2+) transients and inward current, activated by carbachol-induced sarcoplasmic reticulum Ca(2+)release, were not affected by Rho-kinase inhibition. Rho-kinase inhibition produced a Ca(2+)-independent increase in the relaxation rate of contraction, associated with acceleration of MLC dephosphorylation, which was sensitive to calyculin A. These data show for the first time that: (1) Rho-kinase has major effects on Ca(2+) signalling associated with the action potential, (2) this effect is species dependent and (3) Rho-kinase controls relaxation of phasic contraction of myogenic origin. Thus Rho-kinase can modulate phasic smooth muscle in the absence of agonist, and the mechanisms are both Ca(2+)-dependent, involving ion channels, and Ca(2+)-independent, involving MLC phosphorylation activity.


Object-Oriented Segmentation of Cell Nuclei in Fluorescence Microscopy Images.
Cell nucleus segmentation remains an open and challenging problem especially to segment nuclei in cell clumps. Splitting a cell clump would be straightforward if the gradients of boundary pixels in-between the nuclei were always higher than the others. However, imperfections may exist: inhomogeneities of pixel intensities in a nucleus may cause to define spurious boundaries whereas insufficient pixel intensity differences at the border of overlapping nuclei may cause to miss some true boundary pixels. In contrast, these imperfections are typically observed at the pixel-level, causing local changes in pixel values without changing the semantics on a large scale. In response to these issues, this article introduces a new nucleus segmentation method that relies on using gradient information not at the pixel level but at the object level. To this end, it proposes to decompose an image into smaller homogeneous subregions, define edge-objects at four different orientations to encode the gradient information at the object level, and devise a merging algorithm, in which the edge-objects vote for subregion pairs along their orientations and the pairs are iteratively merged if they get sufficient votes from multiple orientations. Our experiments on fluorescence microscopy images reveal that this high-level representation and the design of a merging algorithm using edge-objects (gradients at the object level) improve the segmentation results.


Detection of pediatric respiratory and diarrheal outbreaks from sales of over-the-counter electrolyte products.
To determine whether sales of electrolyte products contain a signal of outbreaks of respiratory and diarrheal disease in children and, if so, how much earlier a signal relative to hospital diagnoses. Retrospective analysis was conducted of sales of electrolyte products and hospital diagnoses for six urban regions in three states for the period 1998 through 2001. Presence of signal was ascertained by measuring correlation between electrolyte sales and hospital diagnoses and the temporal relationship that maximized correlation. Earliness was the difference between the date that the exponentially weighted moving average (EWMA) method first detected an outbreak from sales and the date it first detected the outbreak from diagnoses. The coefficient of determination (r2) measured how much variance in earliness resulted from differences in sales' and diagnoses' signal strengths. The correlation between electrolyte sales and hospital diagnoses was 0.90 (95% CI, 0.87-0.93) at a time offset of 1.7 weeks (95% CI, 0.50-2.9), meaning that sales preceded diagnoses by 1.7 weeks. EWMA with a nine-sigma threshold detected the 18 outbreaks on average 2.4 weeks (95% CI, 0.1-4.8 weeks) earlier from sales than from diagnoses. Twelve outbreaks were first detected from sales, four were first detected from diagnoses, and two were detected simultaneously. Only 26% of variance in earliness was explained by the relative strength of the sales and diagnoses signals (r2 = 0.26). Sales of electrolyte products contain a signal of outbreaks of respiratory and diarrheal diseases in children and usually are an earlier signal than hospital diagnoses.


Positron emission tomography (PET) for assessing aerosol deposition of orally inhaled drug products.
The topical distribution of inhaled therapies in the lung can be viewed using radionuclides and imaging. Positron emission tomography (PET) is a three-dimensional functional imaging technique providing quantitatively accurate localization of the quantity and distribution of an inhaled or injected PET radiotracer in the lung. A series of transaxial slices through the lungs are obtained, comparable to an X-ray computed tomography (CT) scan. Subsequent reformatting allows coronal and sagittal images of the distribution of radioactivity to be viewed. This article describes procedures for administering [(18)F]-fluorodeoxyglucose aerosol to human subjects for the purpose of determining dose and distribution following inhalation from an aerosol drug delivery device (ADDD). The advantages of using direct-labeled PET drugs in the ADDD are discussed with reference to the literature. The methods for designing the inhalation system, determining proper radiation shielding, calibration, and validation of administered radioactivity, scanner setup, and data handling procedures are described. Obtaining an X-ray CT or radionuclide transmission scan to provide accurate geometry of the lung and also correct for tissue attenuation of the PET radiotracer is discussed. Protocols for producing accurate images, including factors that need to be incorporated into the data calibration, are described, as well as a proposed standard method for partitioning the lung into regions of interest. Alternate methods are described for more detailed assessments. Radiation dosimetry/risk calculations for the procedures are appended, as well as a sample data collection form and spreadsheet for calculations. This article should provide guidance for those interested in using PET to determine quantity and distribution of inhaled therapeutics.


Role of human premotor dorsal region in learning a conditional visuomotor task.
Conditional learning is an important component of our everyday activities (e.g., handling a phone or sorting work files) and requires identification of the arbitrary stimulus, accurate selection of the motor response, monitoring of the response, and storing in memory of the stimulus-response association for future recall. Learning this type of conditional visuomotor task appears to engage the premotor dorsal region (PMd). However, the extent to which PMd might be involved in specific or all processes of conditional learning is not well understood. Using transcranial magnetic stimulation (TMS), we demonstrate the role of human PMd in specific stages of learning of a novel conditional visuomotor task that required subjects to identify object center of mass using a color cue and to apply appropriate torque on the object at lift onset to minimize tilt. TMS over PMd, but not vertex, increased error in torque exerted on the object during the learning trials. Analyses of digit position and forces further revealed that the slowing in conditional visuomotor learning resulted from impaired monitoring of the object orientation during lift, rather than stimulus identification, thus compromising the ability to accurately reduce performance error across trials. Importantly, TMS over PMd did not alter production of torque based on the recall of learned color-torque associations. We conclude that the role of PMd for conditional learning is highly sensitive to the stage of learning visuomotor associations. Conditional learning involves stimulus identification, motor response selection, response monitoring, memory encoding, and recall of the learned association. Premotor dorsal (PMd) has been implicated for conditional learning. However, the extent to which PMd might be involved in specific or all stages of conditional learning is not well understood. The novel finding of our study is that PMd appears to be involved with monitoring motor responses, a sensorimotor integration stage essential for conditional learning.


Distinct conditions support a novel classification for bradykinin-mediated angio-oedema.
Angio-oedema (AO) can be attributable to bradykinin (BK) accumulation, as is the case for prototypical hereditary AO (HAO) due to C1 inhibitor (C1-INH) deficiency. However, our clinical experience in a reference centre has shown that some patients display a clinical history suggestive of HAO, but exhibit normal C1-INH function, have no mutation in the causative genes associated with HAO (SERPING1, F12), and report no intake of drugs known to promote AO. We sought to determine the frequency and distribution of different AO subtypes suspected to be BK-mediated AO (BK-AO) and defined by clinical, history and biological criteria (enzyme activities implicated in BK formation and catabolism). The files of all patients referred to our centre for suspected BK-AO were retrospectively analysed. The distribution of patients (n = 162) was 16 and 4% with a hereditary deficiency of C1-INH or a gain of factor XII function, respectively, 29% with iatrogenic BK-AO, 21% with non-iatrogenic defective kininase activity and 30% with idiopathic increased kinin formation. BK-AO may be caused by multiple inherited or acquired factors triggering BK accumulation. Therefore, we propose a novel typology for BK-AO based on the imbalance of production/catabolism of BK.


Static structural signatures of nearly jammed disordered and ordered hard-sphere packings: Direct correlation function.
The nonequilibrium process by which hard-particle systems may be compressed into disordered, jammed states has received much attention because of its wide utility in describing a broad class of amorphous materials. While dynamical signatures are known to precede jamming, the task of identifying static structural signatures indicating the onset of jamming have proven more elusive. The observation that compressing hard-particle packings towards jamming is accompanied by an anomalous suppression of density fluctuations (termed "hyperuniformity") has paved the way for the analysis of jamming as an "inverted critical point" in which the direct correlation function c(r), rather than the total correlation function h(r), diverges. We expand on the notion that c(r) provides both universal and protocol-specific information as packings approach jamming. By considering the degree and position of singularities (discontinuities in the nth derivative) as well as how they are changed by the convolutions found in the Ornstein-Zernike equation, we establish quantitative statements about the structure of c(r) with regards to singularities it inherits from h(r). These relations provide a concrete means of identifying features that must be expressed in c(r) if one hopes to reproduce various details in the pair correlation function accurately and provide stringent tests on the associated numerics. We also analyze the evolution of systems of three-dimensional monodisperse hard spheres of diameter D as they approach ordered and disordered jammed configurations. For the latter, we use the Lubachevsky-Stillinger (LS) molecular dynamics and Torquato-Jiao (TJ) sequential linear programming algorithms, which both generate disordered packings, but can show perceptible structural differences. We identify a short-ranged scaling c(r)∝-1/r as r→0 that accompanies the formation of the delta function at c(D) that indicates the formation of contacts in all cases, and show that this scaling behavior is, in this case, a consequence of the growing long rangedness in c(r), e.g., c∝-1/r^{2} as r→∞ for disordered packings. At densities in the vicinity of the freezing density, we find striking qualitative differences in the structure factor S(k) as well as c(r) between TJ- and LS-generated configurations, including the early formation of a delta function at c(D) in the TJ algorithm's packings, indicating the early formation of clusters of particles in near contact. Both algorithms yield structure factors that tend towards zero in the low-wave-number limit as jamming is approached. Correspondingly, we observe the expected power-law decay in c(r) for large r, in agreement with previous theoretical work. Our work advances the notion that static signatures are exhibited by hard-particle packings as they approach jamming and underscores the utility of the direct correlation function as a sensitive means of monitoring for the appearance of an incipient rigid network.


Do all hemolytic anemias that occur after mitral valve repair require surgical treatment?
We report on a 29-year-old woman with severe hemolytic anemia following mitral valve annuloplasty. Although hemolysis due to mechanical prosthetic mitral valve is well recognized, hemolytic anemia associated with mitral valve repair is an uncommon condition. Reoperation may be considered if the patient has serious and persistent anemia. Although valve replacement is suggested to be a unique intervention, it may not be the solution every time because of mechanical effects. Various mechanisms of hemolysis related to mitral valve repair were suggested, but sufficient and precise data is not available. In this case, we tried to emphasize whether all hemolytic anemias that occur after mitral valve repair require surgical treatment.


Compliance with herpes zoster vaccination in young and adult individuals in two regions of Italy.
The purpose of this work was to explore the knowledge and acceptance of Varicella Zoster Virus (VZV)-Herpes Zoster (HZ) vaccination in the general Italian population, where the HZ vaccine has not yet been distributed, using a prevalence study of subjects from two regions in Italy. A group of 3,173 individuals were interviewed using a questionnaire. The youngest age group (<or= 20 year) was composed of students interviewed at university. The middle age group (21-40 years) and the older age group (>or= 41 years) were interviewed by general practitioners in their office. In both regions, the majority of subjects had been infected with varicella, and only 165 (5.2%) subjects reported receiving the VZV vaccination. Regarding HZ, 2,749 (86.6%) individuals stated that they knew of the virus and 2,233 (70%) were willing to be vaccinated against HZ. The majority of people willing to be vaccinated were in the middle and older age groups (36.6% and 44.7%, respectively). Compliance versus vaccination results were satisfactory and probably, with the upcoming availability of the HZ vaccine in Italy, adults will be favourably disposed towards vaccination.


Effect of corticosteroids on the biomechanical strength of rat rotator cuff tendon.
The effect of corticosteroids on tendon properties is poorly understood, and current data are contradictory and diverse. The biomechanical effect of steroids on rotator cuff tendon has not been studied, to our knowledge. The current study was undertaken to characterize the biomechanical effects of corticosteroid exposure on both uninjured and injured rat rotator cuff tendon. One hundred and twenty-three male Sprague-Dawley rats were randomly assigned to four groups: control (C), tendon injury (I), steroid exposure (S), and tendon injury plus steroid exposure (I+S). Unilateral tendon injuries consisting of a full-thickness defect across 50% of the total width of the infraspinatus tendon were created. Steroid treatment consisted of a single dose of methylprednisolone placed into the subacromial space. At one, three, and five weeks postoperatively, the shoulders were harvested and the infraspinatus tendon was subjected to biomechanical testing. Two specimens from each group were used for histological analysis. At one week, maximum load, maximum stress, and stiffness were all significantly decreased in Group S compared with the values in Group C. Mean maximum load decreased from 37.9 N in Group C to 27.5 N in Group S (p < 0.0005). Mean maximum stress decreased from 18.1 MPa in Group C to 13.6 MPa in Group S (p < 0.0005). Mean stiffness decreased from 26.3 N/mm in Group C to 17.8 N/mm in Group S (p < 0.0005). At one week, mean maximum stress in Group I+S (17.0 MPa) was significantly decreased compared with the value in Group I (19.5 MPa) (p < 0.0005). At both the three-week and the five-week time point, there were no significant differences between Group C and Group S or between Group I and Group I+S with regard to mean maximum load, maximum stress, or stiffness. Histological analysis showed fat cells and collagen attenuation in Groups S and I+S. These changes appeared to be transient. A single dose of corticosteroids significantly weakens both intact and injured rat rotator cuff tendons at one week. This effect is transient as the biomechanical properties of the steroid-exposed groups returned to control levels by three weeks.


Synthesis and photophysical properties of extended π conjugated naphthalimides.
A series of π conjugated naphthalimide derivatives having an imide group as an acceptor conjugated with a methoxy arylethynyl or a methoxyphenyl triazole as a donor were prepared by Sonogashira coupling or "click" chemistry. Their photophysical properties were investigated by steady state and time resolved fluorescence spectroscopy and modelled by TD-DFT calculations. Compound Naphth-AlkyneOMe has a high fluorescence quantum yield and displays efficient photoinduced charge transfer in solution as well as in the powder state. Compound Naphth-TriazoleOMe exhibits a very high Stokes shift and its fluorescence quantum yield is low, which can be rationalized by theoretical calculations.


Antidepressant treatment for acute bipolar depression: an update.
While studies in the past have focused more on treatment of the manic phase of bipolar disorder (BD), recent findings demonstrate the depressive phase to be at least as debilitating. However, in contrast to unipolar depression, depression in bipolar patients exhibits a varying response to antidepressants, raising questions regarding their efficacy and tolerability. Methods. We conducted a MEDLINE and Cochrane Collaboration Library search for papers published between 2005 and 2011 on the subject of antidepressant treatment of bipolar depression. Sixty-eight articles were included in the present review. Results. While a few studies did advocate the use of antidepressants, most well-controlled studies failed to show a robust effect of antidepressants in bipolar depression, regardless of antidepressant class or bipolar subtype. There was no significant increase in the rate of manic/hypomanic switch, especially with concurrent use of mood stabilizers. Prescribing guidelines published in recent years rely more on atypical antipsychotics, especially quetiapine, as a first-line therapy. Conclusions. Antidepressants probably have no substantial role in acute bipolar depression. However, in light of conflicting results between studies, more well-designed trials are warranted.


The emergence of methicillin-resistant Staphylococcus aureus infections in United States hospitals. Possible role of the house staff-patient transfer circuit.
Infections with methicillin-resistant strains of Staphylococcus aureus appear to be occurring with increasing frequency in some U.S. hospitals about a decade after a similar increase in Britain and other countries. In the United States, clustered methicillin-resistant S. aureus infections reported in scientific journals and in three hospital surveys have been almost entirely in large, tertiary referral hospitals affiliated with medical schools. Among 63 hospitals regularly reporting infections from 1974 to 1981 in the National Nosocomial Infections Study, the increase in methicillin-resistant S. aureus infections was entirely due to substantial increases in only four hospitals, all of which were large, tertiary referral centers affiliated with medical schools. The predominance of methicillin-resistant S. aureus infections in these large hospitals may be due to the large numbers of patients at high risk of infection and to the interhospital spread of the organism by the transfer of infected patients and house staff from similar hospitals or from nursing homes.


Sexual dimorphism in the effect of nonhabituating stress on neurogenic plasma extravasation.
The sympathoadrenal axis contributes to the sexual dimorphism of the inflammatory response. As stress both activates the sympathoadrenal axis and profoundly affects inflammation and inflammatory disease, we evaluated whether stress exerts a sexually dimorphic effect on a major component of the inflammatory response, plasma extravasation. We evaluated the effect of a nonhabituating stress, repeated intermittent sound (30 min/day for 4 days), on neurogenic synovial plasma extravasation, induced by bradykinin in the rat knee joint. Sound stress profoundly inhibited bradykinin-induced plasma extravasation in male rats, but profoundly enhanced it in female rats. These effects took 24 h to fully develop after the last exposure to stress. In gonadectomized males, bradykinin-induced plasma extravasation was lower than intact males, and sound stress now enhanced it, i.e. gonadectomized males were phenotypically like intact females. In gonadectomized females, bradykinin-induced plasma extravasation was greater than in intact adult females, and sound stress still enhanced it. Adrenal enucleation significantly attenuated the effect of sound stress on bradykinin-induced plasma extravasation in both male and female rats. We tested the hypothesis that these effects of sound stress were due to sustained enhanced plasma levels of stress hormones. Corticosterone and epinephrine, only when administered in combination, over five days, produced a qualitatively similar effect as sound stress, i.e. bradykinin-induced plasma extravasation was significantly decreased in males and increased in females. These findings suggest that a combined effect of the hypothalamic-pituitary adrenal and sympathoadrenal stress axes are responsible for the marked sexual dimorphism in the effect of stress on the inflammatory response.


Modulation of the CO2 fixation in dinickel azacryptands.
While bimetallic azacryptands are known to selectively coordinate CO2, there is little knowledge on how different substitution patterns of the azacryptand cage structure influence CO2 coordination. Stopped-flow UV-vis spectroscopy, electrochemical analysis and DFT calculations were performed on a series of dinickel azacryptands and showed different rates of CO2 coordination to the complexes. We herein present data showing that the different flexibility of the azacryptands is directly responsible for the difference in the CO2 uptake capability of dinickel azacryptand complexes.


The endocannabinoid system in neurodegeneration.
Endocannabinoids are bioactive lipids, that comprise amides, esters and ethers of long chain polyunsaturated fatty acids. Anandamide (N-arachidonoylethanolamine; AEA) and 2-arachidonoylglycerol (2-AG) are the best studied endocannabinoids, and act as agonists of cannabinoid receptors. Thus, AEA and 2-AG mimic several pharmacological effects of the exogenous cannabinoid delta9-tetrahydrocannabinol, the psychoactive principle of hashish and marijuana. It is known that the activity of endocannabinoids at their receptors is limited by cellular uptake through specific membrane transporters, followed by intracellular degradation by a fatty acid amide hydrolase (for AEA and partly 2-AG) or by a monoacylglycerol lipase (for 2-AG). Together with AEA, 2-AG and congeners, the proteins that bind, transport and metabolize these lipids form the "endocannabinoid system". This new system will be briefly presented in this review, in order to put in a better perspective the role of the endocannabinoid pathway in neurodegenerative disorders, like Parkinson's disease, Huntington's disease, and multiple sclerosis. In addition, the potential exploitation of antagonists of endocannabinoid receptors, or of inhibitors of endocannabinoid metabolism, as next-generation therapeutics will be discussed.


Risk Factors Related to Peripherally Inserted Central Venous Catheter Nonselective Removal in Neonates.
We aimed to investigate the incidence and risk factors associated with nonselective removal of peripherally inserted central venous catheter (PICC) in neonates. In this prospective cohort study, neonates who underwent PICC placement at neonatal intensive care units (NICUs) in China from October 2012 to November 2015 were included. The patient demographics, catheter characteristics, catheter duration, PICC insertion site, indication for PICC insertion, infuscate composition, PICC tip location, and catheter complications were recorded in a computerized database. Risk factors for nonselective removal were analyzed. A total of 497 PICCs were placed in 496 neonates. Nonselective removal occurred in 9.3% of PICCs during 10,540 catheter-days (4.6 nonselective removals per 1,000 catheter-days). These included occlusion (3%), infection (1.4%), leakage (2.0%), phlebitis (0.6%), displacement (1%), pleural effusion(0.6%), and breaks (0.6%). Noncentral tip position was independently associated with an increased risk of nonselective removal (odds ratio 2.621; 95% confidence interval, 1.258-5.461) after adjusting for gestational age, sex, birth weight, and PICC dwell time. No significant differences in the rate of complications occurred between silastic and polyurethane PICC or different insertion sites. Noncentral PICC tip position was the only independent risk factor for nonselective removal of PICC.


Follicular steroid hormones as markers of oocyte quality and oocyte development potential.
Various components of follicular fluid are suggested as biochemical predictors of oocyte quality. Previous studies of follicular steroid hormone levels have shown disparate results when related with fertilization outcomes. The objective of the study was to relate the levels of steroid hormones of each individual follicle with oocyte maturation, fertilization results, embryo quality, and pregnancy rates. Prospective cohort study in a university hospital. In 31 patients, who underwent intracytoplasmic sperm injection, it was performed an ultrasound guided aspiration of follicular fluid of the first two mature follicles from each ovary. Follicular levels of estradiol, progesterone, testosterone, and dehydroepiandrosterone sulfate were measured by chemiluminescent immunoassay. Generalized estimating equation model. In follicular fluids with mature oocyte presence, in normal as well as in failed fertilization, there was a positive correlation between follicular testosterone and progesterone (r = 0.794, P = 0.0001 and r = 0.829, P = 0.0001). Progesterone levels were higher in cases of normal fertilization compared to failed fertilization (P = 0.003). B quality embryos came from oocytes immersed in follicular fluids with higher estradiol values and higher estradiol/progesterone and estradiol/testosterone ratios than those of C quality (P = 0.01; P = 0.0009; P = 0.001). Estradiol levels were higher in patients who achieved pregnancy (P = 0.02). The analysis of follicular hormone composition could be considered as an additional tool in oocyte selection.


Influence of Paleolithic range contraction, admixture and long-distance dispersal on genetic gradients of modern humans in Asia.
Cavalli-Sforza and coauthors originally explored the genetic variation of modern humans throughout the world and observed an overall east-west genetic gradient in Asia. However, the specific environmental and population genetics processes causing this gradient were not formally investigated and promoted discussion in recent studies. Here we studied the influence of diverse environmental and population genetics processes on Asian genetic gradients and identified which could have produced the observed gradient. To do so, we performed extensive spatially-explicit computer simulations of genetic data under the following scenarios: (a) variable levels of admixture between Paleolithic and Neolithic populations, (b) migration through long-distance dispersal (LDD), (c) Paleolithic range contraction induced by the last glacial maximum (LGM), and (d) Neolithic range expansions from one or two geographic origins (the Fertile Crescent and the Yangzi and Yellow River Basins). Next, we estimated genetic gradients from the simulated data and we found that they were sensible to the analysed processes, especially to the range contraction induced by LGM and to the number of Neolithic expansions. Some scenarios were compatible with the observed east-west genetic gradient, such as the Paleolithic expansion with a range contraction induced by the LGM or two Neolithic range expansions from both the east and the west. In general, LDD increased the variance of genetic gradients among simulations. We interpreted the obtained gradients as a consequence of both allele surfing caused by range expansions and isolation by distance along the vast east-west geographic axis of this continent.


Prediction of the recurrence of atrial fibrillation after cardioversion in the Atrial Fibrillation Follow-up Investigation of Rhythm Management (AFFIRM) study.
The early recurrence of atrial fibrillation (AF) after cardioversion and the need for frequent cardioversions to maintain sinus rhythm are important clinical features of AF management. We evaluated patients in the AFFIRM study whose qualifying episode of AF lasted > or = 48 hours and was terminated by cardioversion. Clinical, electrocardiographic, and echocardiographic risk factors associated with AF recurrence within 2 months of cardioversion and > or = 2 cardioversions during the first year were identified using multivariate analysis in 1293 eligible patients. The risk factors for the recurrence of AF within 2 months of cardioversion were no coronary artery disease and an electrocardiographic lead II P-wave duration of > 135 milliseconds. In the subset of patients not taking antiarrhythmic drug therapy, the multivariate risk factors were no coronary artery disease, second or greater episode of AF, left ventricular ejection fraction < 0.50, and mitral valve thickening. Significant risk factors for the need for > 2 cardioversions in the first year in patients taking antiarrhythmic medication were left atrial diameter > 4.5 cm and mitral valve thickening. The overall sensitivity and specificity of these parameters for recurrence and repeated cardioversion are low. There are several risk factors for difficulty maintaining sinus rhythm after cardioversion of persistent AF. The clinical predictive value of these factors is low, and they probably should not be used to justify withholding rhythm control efforts in patients who might benefit from sinus rhythm.


Sports concussion: management and predictors of outcome.
Interest in sports concussion has grown widely in the last two decades among laypersons and medical professionals. Significant contributions of evidence-based research have led to a better understanding of this multifaceted, but still often elusive, injury. This information has transformed all aspects of concussion management, from on-field evaluation through return-to-play guidelines. The aim of this article is to highlight important research regarding predictors of outcome and treatment protocols. This research has been the basis of the paradigm shift from traditional concussion grading scales to individualized care. Today, concussion management requires a patient-centered approach with individualized assessment, including risk factor analysis, neurocognitive testing, and a thorough symptom evaluation.


The influence of geographic variation in socio-cultural factors on length of work disability.
Previous research has largely concentrated on how individual-level factors explain work disability duration. However, growing evidence suggests socio-cultural factors may significantly influence length of work disability. This study examined whether socio-cultural factors that vary by location of residence influence length of disability. This study utilized 216,162 non-work-related disability claims from a private US insurance company and matched these data with socio-cultural factors at the census tract level. OLS regression was used to model findings. Higher unemployment rates, greater median household income, increased poverty status, increased length of the work commute, lower educational attainment, lower percentage of residents living alone, higher percentage of residents age 55 and older, higher percentage of disabled adults (35-64), a lower percentage of whites, and a larger Hispanic population were associated with a longer length of disability. The findings showed that socio-cultural factors varying by location were associated with the length of disability, suggesting that the socioeconomic context of the areas in which individuals reside are likely to influence the work disability process.


Exploring osteoporosis sufferers knowledge on sedentary behaviour in the management of their disease.
1) To develop an understanding of the thoughts and opinions of older women diagnosed with osteoporosis regarding sedentary behaviour and 2) Investigate strategies used to reduce sedentary behaviour for future intervention development. Eleven older women with osteoporosis (mean age=68.2y±6.6(SD)) participated in semi-structured interviews (March-May 2020). They were recruited from the Royal Osteoporosis Society (Scottish) support group networks and the Strathclyde Age-Friendly-Academy. Telephone interviews were recorded, transcribed verbatim and thematically analysed using Braun & Clarke (2006). Three main themes emerged: 'Older Women's Knowledge', 'Motivators to reduce Sedentary Behaviour' and 'Older Adult's and Technology'. Participants reported an increase/maintenance of physical activity levels after osteoporosis diagnosis, had a good understanding and awareness of sedentary behaviour and how it affects health holistically. Participants identified motivators to interrupt sedentary behaviour (e.g. family/friends) and facilitators of sedentary behaviour (e.g. Television). Technology appeared to be used widely among participants to track movement patterns (e.g. Fitbit) but access and usability were identified as potential barriers when using technology to reduce sedentary behaviour among older adults. Knowledge does not appear to be a factor that needs addressing in relation to sedentary behavior in older women diagnosed with osteoporosis. Identified motivators and barriers could increase awareness of sedentary behaviour among older adults.


Metaphase II oocytes from human unilaminar follicles grown in a multi-step culture system.
Can complete oocyte development be achieved from human ovarian tissue containing primordial/unilaminar follicles and grown in vitro in a multi-step culture to meiotic maturation demonstrated by the formation of polar bodies and a Metaphase II spindle? Development of human oocytes from primordial/unilaminar stages to resumption of meiosis (Metaphase II) and emission of a polar body was achieved within a serum free multi-step culture system. Complete development of oocytes in vitro has been achieved in mouse, where in vitro grown (IVG) oocytes from primordial follicles have resulted in the production of live offspring. Human oocytes have been grown in vitro from the secondary/multi-laminar stage to obtain fully grown oocytes capable of meiotic maturation. However, there are no reports of a culture system supporting complete growth from the earliest stages of human follicle development through to Metaphase II. Ovarian cortical biopsies were obtained with informed consent from women undergoing elective caesarean section (mean age: 30.7 ± 1.7; range: 25-39 years, n = 10). Laboratory setting. Ovarian biopsies were dissected into thin strips, and after removal of growing follicles were cultured in serum free medium for 8 days (Step 1). At the end of this period secondary/multi-laminar follicles were dissected from the strips and intact follicles 100-150 μm in diameter were selected for further culture. Isolated follicles were cultured individually in serum free medium in the presence of 100 ng/ml of human recombinant Activin A (Step 2). Individual follicles were monitored and after 8 days, cumulus oocyte complexes (COCs) were retrieved by gentle pressure on the cultured follicles. Complexes with complete cumulus and adherent mural granulosa cells were selected and cultured in the presence of Activin A and FSH on membranes for a further 4 days (Step 3). At the end of Step 3, complexes containing oocytes >100 μm diameter were selected for IVM in SAGE medium (Step 4) then fixed for analysis. Pieces of human ovarian cortex cultured in serum free medium for 8 days (Step 1) supported early follicle growth and 87 secondary follicles of diameter 120 ± 6 μm (mean ± SEM) could be dissected for further culture. After a further 8 days, 54 of the 87 follicles had reached the antral stage of development. COCs were retrieved by gentle pressure from the cultured follicles and those with adherent mural granulosa cells (n = 48) were selected and cultured for a further 4 days (Step 3). At the end of Step 3, 32 complexes contained oocytes >100 μm diameter were selected for IVM (Step 4). Nine of these complexes contained polar bodies within 24 h and all polar bodies were abnormally large. Confocal immuno-histochemical analysis showed the presence of a Metaphase II spindle confirming that these IVG oocytes had resumed meiosis but their developmental potential is unknown. This is a small number of samples but provides proof of concept that complete development of human oocytes can occur in vitro. Further optimization with morphological evaluation and fertilization potential of IVG oocytes is required to determine whether they are normal. The ability to develop human oocytes from the earliest follicular stages in vitro through to maturation and fertilization would benefit fertility preservation practice. Funded by MRC Grants (G0901839 and MR/L00299X/1). No competing interests.


School sports and identity formation: socialisation or selection?
It seems common knowledge that school sport participation leads to all kinds of social, educational and health outcomes. However, it may also be that students with a certain predisposition, sometimes referred to as sporting habitus, are more inclined to participate in school sports and that the 'outcomes' were already present before participation. Several studies indicated that identity formation mediates between sport participation and the outcomes described. Therefore, a longitudinal survey study was used to investigate whether participation in an elementary school sport competition brought about changes in the formation of sport identity and student identity of students. The results of the study showed that participation in the competition was not related to changes in the sport identity and student identity of the children. In contrast to commonplace assumptions about the socialising effects of school sport participation, the results indicate that participating in this school sport competition did not influence the student identity and sport identity of children. It may be that a selected, predisposed group of children with a strong sport identity participates in school sports, although future research is necessary to test this hypothesis.


Advanced spatial knowledge of target location eliminates age-related differences in early sensorimotor learning.
Motor learning has been shown to decline in healthy aging, particularly in the early stages of acquisition. There is now ample evidence that motor learning relies on multiple interacting learning processes that operate on different timescales, but the specific cognitive mechanisms that contribute to motor learning remain unclear. Working memory resources appear to be particularly important during the early stages of motor learning, and declines in early motor learning have been associated with working memory performance in older adults. We examined whether age differences in the early stages of motor learning could be reduced or eliminated by reducing the spatial working memory demands during a force-field adaptation task. Groups of younger and older adults made center-out reaching movements to spatial targets either in a repeating four-element sequence, or in a random order. Participants also performed a battery of cognitive tests to further investigate the potential involvement of associative memory, spatial working memory, and procedural learning mechanisms in the early stage of motor learning. Although all groups adapted their movements equally well by the end of the learning phase, older adults only adapted as quickly as younger adults in the sequence condition, with the older adults in the random group exhibiting slower learning in the earliest stage of motor learning. Across all participants, early motor learning performance was correlated with recognition memory performance on an associative memory test. Within the younger random group, who were able to adapt as quickly as the sequence groups, early motor learning performance was also correlated with performance on a test of procedural learning. These findings suggest that age differences in early stages of motor learning can be eliminated if the spatial working memory demands involved in a motor learning task are limited. Moreover, the results suggest that multiple cognitive resources may be utilized during the early stage of learning, and younger adults may be more flexible than older adults in the recruitment of additional cognitive resources to support learning when spatial working memory demands are high.


Inducing tolerance to a soluble foreign antigen by encapsulated cell transplants.
The immune response to soluble antigens constitutes a current clinical problem impeding the development of protein therapeutics. We have developed an encapsulated-cell delivery system, which, transiently combined with an anti-CD154 antibody treatment, allows for the suppression of this immune response and the establishment of long-term secretion of a foreign antigen, human erythropoietin (huEPO). The chronic presence of antigen appears to be required to maintain this tolerance, as a 21-day gap in the exposure to huEPO is sufficient to restore the ability of mice to mount an antibody response. In contrast, chronic huEPO expression maintains tolerance even in the absence of further anti-CD154 treatment. These results suggest that a soluble antigenic protein can be continuously released, without inducing an antibody response, using encapsulated allogeneic cells. The temporary anti-CD154 treatment induces immune unresponsiveness to the delivered antigen, while the immunoprotected cell implant allows for chronic antigen release, favoring the establishment of tolerance.


Parental acceptance of HPV vaccine in Peru: a decision framework.
Cervical cancer is the third most common cancer affecting women worldwide and it is an important cause of death, especially in developing countries. Cervical cancer is caused by human papillomavirus (HPV) and can be prevented by HPV vaccine. The challenge is to expand vaccine availability to countries where it is most needed. In 2008 Peru's Ministry of Health implemented a demonstration project involving 5(th) grade girls in primary schools in the Piura region. We designed and conducted a qualitative study of the decision-making process among parents of girls, and developed a conceptual model describing the process of HPV vaccine acceptance. We found a nonlinear HPV decision-making process that evolved over time. Initially, the vaccine's newness, the requirement of written consent, and provision of information were important. If information was sufficient and provided by credible sources, many parents accepted the vaccine. Later, after obtaining additional information from teachers, health personnel, and other trusted sources, more parents accepted vaccination. An understanding of the issues surrounding the vaccine developed, parents overcome fears and rumors, and engaged in family negotiations-including hearing the girl's voice in the decision-making process. The concept of prevention (cancer as danger, future health, and trust in vaccines) combined with pragmatic factors (no cost, available at school) and the credibility of the offer (information in the media, recommendation of respected authority figure) were central to motivations that led parents to decide to vaccinate their daughters. A lack of confidence in the health system was the primary inhibitor of vaccine acceptance. Health personnel and teachers are credible sources of information and can provide important support to HPV vaccination campaigns.


Intranasal oxytocin increases covert attention to positive social cues.
The neuropeptide oxytocin (OT) has positive effects on the processing of emotional stimuli such as facial expressions. To date, research has focused primarily on conditions of overt visual attention. We investigated whether a single intranasal dose of OT (24 IU) would modulate the allocation of attentional resources towards positive and negative facial expressions using a dot-probe paradigm in a sample of 69 healthy men. Attentional capacity for these facial cues was limited by presentation time (100 or 500 ms). In addition, we controlled for overt visual attention by recording eye movements using a remote eye tracker. Reaction times (RTs) in the dot-probe paradigm revealed a pronounced shift of attention towards happy facial expressions presented for 100 ms after OT administration, whereas there were no OT-induced effects for longer presentation times (500 ms). The results could not be attributed to modulations of overt visual attention as no substance effects on gazes towards the facial target were observed. The results suggest that OT increased covert attention to happy faces, thereby supporting the hypothesis that OT modulates early attentional processes that might promote prosocial behavior.


Topoisomerases and Anthracyclines: Recent Advances and Perspectives in Anticancer Therapy and Prevention of Cardiotoxicity.
Topoisomerases are ubiquitous enzymes involved in maintaining genomic stability of the cell by regulating the over- or underwinding of DNA strands. Besides their customary functions, topoisomerases are important cellular targets of widely used anticancer drugs. In particular, topoisomerase IIα (Top2α) has been postulated as the primary molecular target of anthracycline's anticancer activity, whereas topoisomerase IIβ (Top2β), the only Top2 present in heart tissue, seems to be involved in the development of anthracycline-induced cardiotoxicity. Noteworthy, cardiotoxicity is the most frequent adverse effect of both conventional and modern anticancer targeted therapy, representing the leading noncancer-related cause of morbidity and mortality in long-term survivors. The molecular mechanisms of anthracyclineinduced cardiotoxicity have been investigated for decades and, despite the numerous mechanistic hypotheses put forward, its aetiology and pathogenesis still remain controversial. This review is aimed at focusing on the double edge sword of topoisomerase-anthracycline interaction, and, in particular, on the potential role of topoisomerases in anthracyclines anticancer activity as well as in the pathogenesis of anthracycline-induced cardiotoxicity.


Contrast magnetic resonance imaging for measurement of cartilage glycosaminoglycan content in dogs: a pilot study.
To assess the ability of a contrast-enhanced magnetic resonance imaging (MRI) technique to quantitatively determine glycosaminoglycan content in canine articular cartilage. Fifty-four full-thickness cartilage discs were collected from the femorotibial and scapulohumeral joints of three adult dogs immediately following euthanasia. One set of discs from each dog was analysed for glycosaminoglycan content using a colourimetric laboratory assay. The remaining position-matched set of discs from contralateral limbs underwent pre- and post-contrast gadolinium-enhanced MRI, using repeated saturation recovery pulse sequences which were used to generate calculated T1 maps of the cartilage discs. Linear regression analysis was then performed relating delayed gadolinium-enhanced MRI T1 calculated signal intensity to the cartilage glycosaminoglycan content normalized to DNA content. Repeatability of triplicate measurements was estimated by calculating the coefficient of variation. Mean coefficient of variation estimates for the gadolinium-enhanced MRI T1 signal intensity values for nine sampling sites from three dogs ranged from 5.9% to 7.5%. Gadolinium-enhanced MRI T1 signal intensity was significantly correlated (p <0.05) with normalized glycosaminoglycan content in two dogs (r = 0.79, p = 0.011; r = 0.78, p = 0.048), but not in the third dog (r = 0.53, p = 0.071). Gadolinium-enhanced MRI assessment of cartilage may be predictive of glycosaminoglycan content and therefore offer an in vivo assessment of changes in cartilage characteristics over time. Additional studies appear indicated to determine the reliability and clinical applicability of gadolinium-enhanced MRI in detecting changes in cartilage over time.


Eight Wonders of the solar system.
Artist Ron Miller takes us on a journey to eight of the most breathtaking views that await intrepid explorers of our solar system. The scale of these natural wonders dwarfs anything Earth has to offer. What might we see and feel if we could travel to these distant domains? The artist's eye-interpreting data from probes such as NASA's Cassini, which is now exploring the Saturnian system, and MESSENGER, which has flown by Mercury three times and goes into permanent orbit next March-allows us an early visit to these unforgettable locales.


Human and Helicobacter pylori Interactions Determine the Outcome of Gastric Diseases.
The innate immune response is a critical hallmark of Helicobacter pylori infection. Epithelial and myeloid cells produce effectors, including the chemokine CXCL8, reactive oxygen species (ROS), and nitric oxide (NO), in response to bacterial components. Mechanistic and epidemiologic studies have emphasized that dysregulated and persistent release of these products leads to the development of chronic inflammation and to the molecular and cellular events related to carcinogenesis. Moreover, investigations in H. pylori-infected patients about polymorphisms of the genes encoding CXCL8 and inducible NO synthase, and epigenetic control of the ROS-producing enzyme spermine oxidase, have further proven that overproduction of these molecules impacts the severity of gastric diseases. Lastly, the critical effect of the crosstalk between the human host and the infecting bacterium in determining the severity of H. pylori-related diseases has been supported by phylogenetic analysis of the human population and their H. pylori isolates in geographic areas with varying clinical and pathologic outcomes of the infection.


Disorders of higher gaze control.
The neural centers in the cerebral hemispheres, both cortex and basal ganglia, involved in the generation of saccadic and smooth pursuit eye movements have been well delineated in terms of their location and function. For the generation of saccades these include the frontal eye fields, the supplementary eye field, and the intraparietal sulcus, and in the basal ganglia the caudate nucleus and the substantia nigra, pars compacta. The generation of pursuit eye movements involves the middle temporal (area V5) and medial superior temporal areas and the frontal eye field. These centers and their connections are disturbed not only in acute and chronic lesions such as cerebral infarction, but also in a wide variety of neurodegenerative diseases. In certain of these conditions, such as patients with cortical dementias and basal ganglia disorders, correct interpretation of the resulting eye movement abnormalities can contribute to differentiating between a range of differential diagnoses.


Isoliquiritigenin alleviates early brain injury after experimental intracerebral hemorrhage via suppressing ROS- and/or NF-κB-mediated NLRP3 inflammasome activation by promoting Nrf2 antioxidant pathway.
Intracerebral hemorrhage (ICH) induces potently oxidative stress responses and inflammatory processes. Isoliquiritigenin (ILG) is a flavonoid with a chalcone structure and can activate nuclear factor erythroid-2 related factor 2 (Nrf2)-mediated antioxidant system, negatively regulate nuclear factor-κB (NF-κB) and nod-like receptor family, pyrin domain-containing 3 (NLRP3) inflammasome pathways, but its role and potential molecular mechanisms in the pathology following ICH remain unclear. The present study aimed to explore the effects of ILG after ICH and underlying mechanisms. ICH model was induced by collagenase IV (0.2 U in 1 μl sterile normal saline) in male Sprague-Dawley rats weighing 280-320 g. Different doses of ILG (10, 20, or 40 mg/kg) was administrated intraperitoneally at 30 min, 12 h, 24 h, and 48 h after modeling, respectively. Rats were intracerebroventricularly administrated with control scramble small interfering RNA (siRNA) or Nrf2 siRNA at 24 h before ICH induction, and after 24 h, ICH model was established with or without ILG (20 mg/kg) treatment. All rats were dedicated at 24 or 72 h after ICH. Neurological deficits, histological damages, brain water content (BWC), blood-brain barrier (BBB) disruption, and neuronal degeneration were evaluated; quantitative real-time RT-PCR (qRT-PCR), immunohistochemistry/immunofluorescence, western blot, and enzyme-linked immunosorbent assay (ELISA) were carried out; catalase, superoxide dismutase activities and reactive oxygen species (ROS), and glutathione/oxidized glutathione contents were measured. ILG (20 and 40 mg/kg) markedly alleviated neurological deficits, histological damages, BBB disruption, brain edema, and neuronal degeneration, but there was no significant difference between two dosages. ILG (20 mg/kg) significantly suppressed the NF-κB and NLRP3 inflammasome pathways and activated Nrf2-mediated antioxidant system. Gene silencing of Nrf2 aggravated the neurological deficits, brain edema, and neuronal degeneration and increased the protein levels of NF-κB p65, NLRP3 inflammasome components, and IL-1β. ILG delivery significantly attenuated the effects of Nrf2 siRNA interference mentioned above. Intraperitoneal administration of ILG after ICH reduced early brain impairments and neurological deficits, and the mechanisms were involved in the regulation of ROS and/or NF-κB on the activation of NLRP3 inflammasome pathway by the triggering of Nrf2 activity and Nrf2-induced antioxidant system. In addition, our experimental results may make ILG a potential candidate for a novel therapeutical strategy for ICH.


Using simulation modelling to examine the impact of venue lockout and last-drink policies on drinking-related harms and costs to licensees.
Many variations of venue lockout and last-drink policies have been introduced in attempts to reduce drinking-related harms. We estimate the public health gains and licensee costs of these policies using a computer simulated population of young adults engaging in heavy drinking. Using an agent-based model we implemented 1 am/2 am/3 am venue lockouts in conjunction with last drinks zero/one/two hours later, or at current closing times. Outcomes included: the number of incidents of verbal aggression in public drinking venues, private venues or on the street; and changed revenue to public venues. The most effective policy in reducing verbal aggression among agents was 1 am lockouts with current closing times. All policies produced substantial reductions in street-based incidents of verbal aggression among agents (33-81%) due to the smoothing of transport demand. Direct revenue losses were 1-9% for simulated licensees, with later lockout times and longer periods between lockout and last drinks producing smaller revenue losses. Simulation models are useful for exploring consequences of policy change. Our simulation suggests that additional hours between lockout and last drinks could reduce aggression by easing transport demand, while minimising revenue loss to venue owners. Implications for public health: Direct policies to reduce late-night transport-related disputes should be considered.


Genetic editing of HBV DNA by monodomain human APOBEC3 cytidine deaminases and the recombinant nature of APOBEC3G.
Hepatitis B virus (HBV) DNA is vulnerable to editing by human cytidine deaminases of the APOBEC3 (A3A-H) family albeit to much lower levels than HIV cDNA. We have analyzed and compared HBV editing by all seven enzymes in a quail cell line that does not produce any endogenous DNA cytidine deaminase activity. Using 3DPCR it was possible to show that all but A3DE were able to deaminate HBV DNA at levels from 10(-2) to 10(-5)in vitro, with A3A proving to be the most efficient editor. The amino terminal domain of A3G alone was completely devoid of deaminase activity to within the sensitivity of 3DPCR ( approximately 10(-4) to 10(-5)). Detailed analysis of the dinucleotide editing context showed that only A3G and A3H have strong preferences, notably CpC and TpC. A phylogenic analysis of A3 exons revealed that A3G is in fact a chimera with the first two exons being derived from the A3F gene. This might allow co-expression of the two genes that are able to restrict HIV-1Deltavif efficiently.


Oral tolerance in protein-deprived mice. II. Evidence of normal 'gut processing' of ovalbumin, but suppressor cell deficiency, in deprived mice.
The induction of oral tolerance for DTH responses to ovalbumin is impaired in protein-deprived mice. This may be via the effects of protein malnutrition on short-lived Ts cells, but an alternative explanation is that the gut handling of antigen is abnormal. We have attempted to transfer tolerance from protein-deprived and control mice to naive recipients by using spleen cells, collected 7 days after an OVA feed at a time when Ts cells should be present, or by using serum, collected 1 hr after feeding, which should contain tolerogenic, 'gut-processed' antigen. Suppression of DTH was transferred with 7-day spleen cells and with 1-hr serum from normal, protein-sufficient mice. Mice that received spleen cells from protein-deprived donors were not tolerant, but suppression was readily transferred with serum from deprived mice, indicating that their capacity for intestinal antigen processing was normal. Furthermore, the quantity of absorbed antigen in the serum 1 hr after feeding was similar in both protein-deprived and normal groups. The results obtained are consistent with the hypothesis that short-term protein deprivation depletes a population of short-lived Ts cells which control DTH oral tolerance.


The presence of anti-phosphatidylserine/prothrombin antibodies as risk factor for both arterial and venous thrombosis in patients with systemic lupus erythematosus.
In an effort to clarify the clinical significance of anti-phospholipid antibodies (aPL) detected by enzyme-linked immunosorbent assay (ELISA), we examined the prevalence of anti-cardiolipin antibodies (aCL), anti-beta2-glycoprotein I antibodies (anti-beta2-GPI), antiprothrombin antibodies (anti-PT), and anti-phosphatidylserine/prothrombin antibodies (anti-PS/PT) in 175 patients with systemic lupus erythematosus (SLE) comprising 67 patients with thrombotic complications. The present study showed that positive results of anti-beta2-GPI-ELISA and anti-PS/PT-ELISA could serve as markers of thrombotic complications in patients with SLE, whereas aCL and anti-PT are less reliable as markers of these complications. Furthermore, results of the anti-PS/PT-ELISA correlate best with the occurrence of both arterial and venous thrombosis in patients with SLE.


SpaceX: gene co-expression network estimation for spatial transcriptomics.
The analysis of spatially resolved transcriptome enables the understanding of the spatial interactions between the cellular environment and transcriptional regulation. In particular, the characterization of the gene-gene co-expression at distinct spatial locations or cell types in the tissue enables delineation of spatial co-regulatory patterns as opposed to standard differential single gene analyses. To enhance the ability and potential of spatial transcriptomics technologies to drive biological discovery, we develop a statistical framework to detect gene co-expression patterns in a spatially structured tissue consisting of different clusters in the form of cell classes or tissue domains. We develop SpaceX (spatially dependent gene co-expression network), a Bayesian methodology to identify both shared and cluster-specific co-expression network across genes. SpaceX uses an over-dispersed spatial Poisson model coupled with a high-dimensional factor model which is based on a dimension reduction technique for computational efficiency. We show via simulations, accuracy gains in co-expression network estimation and structure by accounting for (increasing) spatial correlation and appropriate noise distributions. In-depth analysis of two spatial transcriptomics datasets in mouse hypothalamus and human breast cancer using SpaceX, detected multiple hub genes which are related to cognitive abilities for the hypothalamus data and multiple cancer genes (e.g. collagen family) from the tumor region for the breast cancer data. The SpaceX R-package is available at github.com/bayesrx/SpaceX. Supplementary data are available at Bioinformatics online.


Mathematical model predicts tumor control patterns induced by fast and slow cytotoxic T lymphocyte killing mechanisms.
Immunotherapy has dramatically transformed the cancer treatment landscape largely due to the efficacy of immune checkpoint inhibitors (ICIs). Although ICIs have shown promising results for many patients, the low response rates in many cancers highlight the ongoing challenges in cancer treatment. Cytotoxic T lymphocytes (CTLs) execute their cell-killing function via two distinct mechanisms: a fast-acting, perforin-mediated process and a slower, Fas ligand (FasL)-driven pathway. Evidence also suggests that the preferred killing mechanism of CTLs depends on the antigenicity of tumor cells. To determine the critical factors affecting responses to ICIs, we construct an ordinary differential equation model describing in vivo tumor-immune dynamics in the presence of active or blocked PD-1/PD-L1 immune checkpoint. Specifically, we identify important aspects of the tumor-immune landscape that affect tumor size and composition in the short and long term. We also generate a virtual cohort of mice with diverse tumor and immune attributes to simulate the outcomes of immune checkpoint blockade in a heterogeneous population. By identifying key tumor and immune characteristics associated with tumor elimination, dormancy, and escape, we predict which fraction of a population potentially responds well to ICIs and ways to enhance therapeutic outcomes with combination therapy.


Molecular pathomechanisms and cell-type-specific disease phenotypes of MELAS caused by mutant mitochondrial tRNA(Trp).
Numerous pathogenic mutations responsible for mitochondrial diseases have been identified in mitochondrial DNA (mtDNA)-encoded tRNA genes. In most cases, however, the detailed molecular pathomechanisms and cellular pathophysiology of these mtDNA mutations -how such genetic defects determine the variation and the severity of clinical symptoms in affected individuals- remain unclear. To investigate the molecular pathomechanisms and to realize in vitro recapitulation of mitochondrial diseases, intracellular mutant mtDNA proportions must always be considered. We found a disease-causative mutation, m.5541C>T heteroplasmy in MT-TW gene, in a patient exhibiting mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) with multiple organ involvement. We identified the intrinsic molecular pathomechanisms of m.5541C>T. This mutation firstly disturbed the translation machinery of mitochondrial tRNA(Trp) and induced mitochondrial respiratory dysfunction, followed by severely injured mitochondrial homeostasis. We also demonstrated cell-type-specific disease phenotypes using patient-derived induced pluripotent stem cells (iPSCs) carrying ~100 % mutant m.5541C>T. Significant loss of terminally differentiated iPSC-derived neurons, but not their stem/progenitor cells, was detected most likely due to serious mitochondrial dysfunction triggered by m.5541C>T; in contrast, m.5541C>T did not apparently affect skeletal muscle development. Our iPSC-based disease models would be widely available for understanding the "definite" genotype-phenotype relationship of affected tissues and organs in various mitochondrial diseases caused by heteroplasmic mtDNA mutations, as well as for further drug discovery applications.