IL-1β-pre-conditioned mesenchymal stem/stromal cells' secretome modulates the inflammatory response and aggrecan deposition in intervertebral disc.
Mesenchymal stem/stromal cells (MSCs) have been increasingly used in clinical trials for low-back pain (LBP) and intervertebral disc (IVD) degeneration with promising results. Their action mechanisms are not fully understood, but they reduce IVD pro-inflammatory markers in a pro-inflammatory/degenerative IVD microenvironment. In this study the therapeutic potential of the MSC secretome, as an alternative cell-free approach for treating degenerated IVDs, was examined. Human bone marrow-derived MSC secretome (MSCsec) was collected after 48 h of preconditioning in IL-1β (10 ng/mL) and low oxygen (6 % O2), mimicking the degenerative IVD. IL-1β-pre-conditioning of MSCs increased secretion of pro-inflammatory markers hIL-6, hIL-8, hMCP-1, etc. The therapeutic effect of MSCsec was tested in a pro-inflammatory/degenerative IVD ex vivo model. MSCsec down-regulated IVD gene expression of pro-inflammatory cytokines (bIL-6, bIL-8) and matrix degrading enzyme bMMP1, while bMMP3 and bTIMP2 were up-regulated, at 48 h. After 14 d, MSCsec-treated IVDs revealed increased aggrecan deposition, although no differences in other ECM components were observed. Protein analysis of the MSCsec-treated IVD supernatant revealed a significant increase of CXCL1, MCP-1, MIP-3α, IL-6, IL-8 and GRO α/β/γ (related to TNF, NOD-like receptor and neutrophil chemotaxis signalling), and a decrease of IFN-γ, IL-10, IL-4, IL-5 and TNF-α (associated with T-cell receptor signalling). MSCsec-treated IVD supernatants did not promote angiogenesis and neurogenesis in vitro. Overall, MSCsec can be a safe therapeutic approach, presenting a strong immunomodulatory role in degenerated IVD while potentiating aggrecan deposition, which can open new perspectives on the use of MSCsec as a cell-based/ cell-free therapeutic approach to LBP.


The FGFR/MEK/ERK/brachyury pathway is critical for chordoma cell growth and survival.
Recent evidence suggests that the expression of brachyury is necessary for chordoma growth. However, the mechanism associated with brachyury-regulated cell growth is poorly understood. Fibroblast growth factor (FGF), a regulator of brachyury expression in normal tissue, may also play an important role in chordoma pathophysiology. Using a panel of chordoma cell lines, we explored the role of FGF signaling and brachyury in cell growth and survival. Western blots showed that all chordoma cell lines expressed fibroblast growth factor receptor 2 (FGFR2), FGFR3, mitogen-activated protein kinase kinase (MEK) and extracellular signal-regulated kinase (ERK), whereas no cell lines expressed FGFR1 and FGFR4. Results of enzyme-linked immunosorbent assay indicated that chordoma cells produced FGF2. Neutralization of FGF2 inhibited MEK/ERK phosphorylation, decreased brachyury expression and induced apoptosis while reducing cell growth. Activation of the FGFR/MEK/ERK/brachyury pathway by FGF2-initiated phosphorylation of FGFR substrate 2 (FRS2)-α (Tyr196) prevented apoptosis while promoting cell growth and epithelial-mesenchymal transition (EMT). Immunofluorescence staining showed that FGF2 promoted the translocation of phosphorylated ERK to the nucleus and increased brachyury expression. The selective inhibition of FGFR, MEK and ERK phosphorylation by PD173074, PD0325901 and PD184352, respectively, decreased brachyury expression, induced apoptosis, and inhibited cell growth and EMT. Moreover, knockdown of brachyury by small hairpin RNA reduced FGF2 secretion, inhibited FGFR/MEK/ERK phosphorylation and blocked the effects of FGF2 on cell growth, apoptosis and EMT. Those findings highlight that FGFR/MEK/ERK/brachyury pathway coordinately regulates chordoma cell growth and survival and may represent a novel chemotherapeutic target for chordoma.


Surface energy analysis as a tool to probe the surface energy characteristics of micronized materials--a comparison with inverse gas chromatography.
This study investigates the impact of micronization on the measured surface energy characteristics of an active pharmaceutical ingredient (API), ibipinabant, by inverse gas chromatography (IGC) using both a fixed probe concentration, commonly used in standard IGC methods, and a fixed probe surface coverage approach applied by the surface energy analyzer (SEA), a next generation IGC system. The IGC measurements indicate an initial increase in surface energy, going from un-micronized to micronized, followed by a reduction in surface energy with increasing micronization extent. This was attributable to the change in the retention behaviour of the dispersive probes as a consequence of the change in the probe surface coverage rather than a change in the actual surface energy of the materials being analysed. It was observed in the SEA data that micronization leads to an increase in the measured dispersive surface energy of the drug substance with increasing micronization extent. The increase in surface energy is primarily due to the generation of new, higher energy interaction sites, although a small additional increase is also observed which is related to the increase in the number and distribution of high energy sites. The results demonstrate that in order to obtain comparable surface energetic data between batches with varied surface area, and presumably between different materials, results should be obtained at a specific, and constant, probe surface coverage.


Breathing biofeedback as an adjunct to exposure in cognitive behavioral therapy hastens the reduction of PTSD symptoms: a pilot study.
Although trauma-focused cognitive behavioral therapy (TF-CBT) with exposure is an effective treatment for posttraumatic stress disorder (PTSD), not all patients recover. Addition of breathing biofeedback to exposure in TF-CBT is suggested as a promising complementary technique to improve recovery of PTSD symptoms. Patients (n = 8) with chronic PTSD were randomized to regular TF-CBT or TF-CBT with complementary breathing biofeedback to exposure. PTSD symptoms were measured before, during and after TF-CBT with the Impact of Event Scale-Revised. The results show that breathing biofeedback is feasible and can easily be complemented to TF-CBT. Although PTSD symptoms significantly decreased from pre to post treatment in both conditions, there was a clear trend towards a significantly faster (p = .051) symptom reduction in biofeedback compared to regular TF-CBT. The most important limitation was the small sample size. The hastened clinical improvement in the biofeedback condition supports the idea that breathing biofeedback may be an effective complementary component to exposure in PTSD patients. The mechanism of action of breathing biofeedback may relate to competing working memory resources decreasing vividness and emotionality, similar to eye movement desensitization and reprocessing. Future research is needed to examine this.


Superiority of amniotic fluid index over amniotic fluid pocket measurement for predicting bad fetal outcome.
Semiquantitative amniotic fluid volume (AFV) determination is a component of the fetal biophysical profile (BPP). To assess decreased AVF we did BPPs of 174 fetuses within 1 week of delivery. Two methods were used: measurement of the single largest vertical pocket (oligohydramnios = depth < 1 cm) and the four-quadrant amniotic fluid index (AFI) (oligohydramnios = AFI < or = 5 cm). AFV, as determined by each method, was related to measures of fetal outcome (perinatal mortality, fetal distress, Apgar score, meconium-stained amniotic fluid, and intrauterine growth retardation [IUGR]). The AFI was more sensitive in predicting mortality (87.5%) and the following measures of perinatal morbidity: low 5-minute Apgar score (88.8%), fetal distress during labor (86.6%), meconium-stained amniotic fluid (63.6%), and the presence of IUGR (79.4%). The sensitivity of amniotic fluid pocket measurement of < 1 cm was 75%, 72.2%, 66.6%, 47.7%, and 55.8%, respectively, for the same measures. Using the AFI instead of a single pocket measurement in BPP assessment increased the sensitivity and positive predictive value of the BPP from 64.7% to 76.4% and from 45.8% to 68.4%, respectively. Our data suggest that qualitative AFV measurement using the AFI is superior to that of the single largest pocket in multiple component fetal biophysical testing.


Artocarpus communis Induces Autophagic Instead of Apoptotic Cell Death in Human Hepatocellular Carcinoma Cells.
For centuries, natural plant extracts have played an important role in traditional medicine for curing and preventing diseases. Studies have revealed that Artocarpus communis possess various bioactivities, such as anti-inflammation, anti-oxidant, and anticancer activities. A. communis offers economic value as a source of edible fruit, yields timber, and is widely used in folk medicines. However, little is known about its molecular mechanisms of anticancer activity. Here, we demonstrate the antiproliferative activity of A. communis methanol extract (AM) and its dichloromethane fraction (AD) in two human hepatocellular carcinoma (HCC) cell lines, HepG2 and PLC/PRF/5. Colony assay showed the long-term inhibitory effect of both extracts on cell growth. DNA laddering and immunoblotting analyses revealed that both extracts did not induce apoptosis in the hepatoma cell lines. AM and AD-treated cells demonstrated different cell cycle distribution compared to UV-treated cells, which presented apoptotic cell death with high sub-G1 ratio. Instead, acridine orange staining revealed that AM and AD triggered autophagosome accumulation. Immunoblotting showed a significant expression of autophagy-related proteins, which indicated the autophagic cell death (ACD) of hepatoma cell lines. This study therefore demonstrates that A. communis AM and its dichloromethane fraction can induce ACD in HCC cells and elucidates the potential of A. communis extracts for development as anti tumor therapeutic agents that utilize autophagy as mechanism in mediating cancer cell death.


[Impact of Streptococcus pneumoniae on pneumonia in Latin American children. SIREVA-Vigía Group].
Community-acquired pneumonia is one of the leading causes of infant morbidity and mortality. Studies conducted in developing countries indicate that the most serious symptoms of pneumonia are associated with bacterial causes, mainly Streptococcus pneumoniae, followed by Haemophilus influenzae type b. Managing those infections in children under two years of age is hindered by the lack of appropriate vaccines and by the decreased susceptibility of S. pneumoniae to penicillin and other antibiotics. In 1993, at the initiative of the Regional System for Vaccines of the Pan American Health Organization, and with funding from the Canadian International Development Agency, a study was designed to identify the S. pneumoniae capsular types that cause invasive disease in Latin American children under 5 years of age. The objective of the study was to determine the ideal composition of a conjugate vaccine that could be used in Latin America, and the penicillin susceptibility of the S. pneumoniae isolates. The initiative was undertaken in Argentina, Brazil, Chile, Colombia, Mexico, and Uruguay. This report analyzes the information that the participating countries generated on pneumococcal pneumonia. A total of 3,393 children were found with systemic S. pneumoniae infections, of which 1,578 corresponded to pneumonias. The analysis focused on 1,409 cases of pneumonia in Argentina, Brazil, Colombia, Mexico, and Uruguay. Of the children, 63.8% of them were under two years of age. Twelve prevalent capsular types were identified, of which serotypes 14, 5, and 1 were the three most common in the majority of the countries. At the beginning of the study the highest level of penicillin resistance was found in Mexico (47.0%), and the lowest in Colombia (12.1%). Over the 1993-1998 period, resistance to penicillin increased in the five countries. Penicillin resistance was associated with a small number of capsular serotypes, mainly 14 and 23F. The first of those serotypes was resistant to penicillin and to trimethoprim-sulfamethoxazole, and the second was multiresistant. The frequency of resistance to trimethoprim-sulfamethoxazole was high in all of the countries; Argentina had the highest level, 58.0%. A decrease in susceptibility to chloramphenicol was uncommon, except in Colombia, where there was a resistance level of 23.4%. Resistance to erythromycin was low in all the countries, and all the isolates were susceptible to vancomycin.


Mortality and Cancer Incidence in Carriers of Balanced Robertsonian Translocations: A National Cohort Study.
A balanced robertsonian translocation (rob) results from fusion of 2 acrocentric chromosomes. Carriers are phenotypically normal and are often diagnosed because of recurrent miscarriages, infertility, or aneuploid offspring. Mortality and site-specific cancer risks in carriers have not been prospectively investigated. We followed 1,987 carriers diagnosed in Great Britain for deaths and cancer risk, over an average of 24.1 years. Standardized mortality and incidence ratios were calculated comparing the number of observed events against population rates. Overall mortality was higher for carriers diagnosed before age 15 years (standardized mortality ratio (SMR) = 2.00, 95% confidence interval (CI): 1.09, 3.35), similar for those diagnosed aged 15-44 years (SMR = 1.06, 95% CI: 0.86-1.28), and lower for those diagnosed aged 45-84 years (SMR = 0.81, 95% CI: 0.68, 0.95). Cancer incidence was higher for non-Hodgkin lymphoma (standardized incidence ratio (SIR) = 1.90, 95% CI: 1.01, 3.24) and childhood leukemia (SIR = 14.5, 95% CI: 1.75, 52.2), the latter particularly in rob(15;21) carriers (SIR = 447.8, 95% CI: 11.3, 2,495). Rob(13;14) carriers had a higher breast cancer risk (SIR = 1.58, 95% CI: 1.12, 2.15). Mortality risks relative to the population in diagnosed carriers depend on age at cytogenetic diagnosis, possibly reflecting age-specific cytogenetic referral reasons. Carriers might be at greater risk of childhood leukemia and non-Hodgkin lymphoma and those diagnosed with rob(13;14) of breast cancer.


Effects of recombinant human erythropoietin in normal humans.
This review describes some of the physiological effects of recombinant human erythropoietin (EPO) in healthy humans. At the blood level EPO increases the arterial O(2) content not only by increasing red blood cell volume, but also by an equally important decrease in plasma volume. Well before that, EPO causes a prompt decrease in plasma levels of renin and aldosterone. Renal clearance studies suggest that EPO decreases renal proximal tubular reabsorption rate leading to activation of the tubuloglomerular feedback mechanism and a fall in glomerular filtration rate. Thus, treatment with EPO may result in suppression of endogenous EPO production through a decrease in intrarenal oxygen consumption. EPO elevates the arterial blood pressure even in healthy subjects. The receptor for EPO is present in many tissues. However, the functional effects of EPO in the skeletal muscle seem limited, and although it has been speculated that non-erythropoietic effects of EPO (angiogenesis, shift in muscle fibre types, cognitive effects) may be responsible for the increase in exercise performance, this has not been confirmed. EPO-induced haemodynamic effects call for careful monitoring during the administration period. The metabolic, hormonal and renal effects of EPO do not seem to range beyond physiologically acceptable limits and are reversible. Taken together, EPO seems safe to use for experimental purposes in healthy volunteers.


Serological responses to prednisolone treatment in leprosy reactions: study of TNF-α, antibodies to phenolic glycolipid-1, lipoarabinomanan, ceramide and S100-B.
Corticosteroids have been extensively used in the treatment of immunological reactions and neuritis in leprosy. The present study evaluates the serological response to steroid treatment in leprosy reactions and neuritis. Seven serological markers [TNF-α, antibodies to Phenolic glycolipid-1 (PGL-1 IgM and IgG), Lipoarabinomannan (LAM IgG1 and IgG3), C2-Ceramide and S100 B] were analyzed longitudinally in 72 leprosy patients before, during and after the reaction. At the onset of reaction these patients received a standard course of prednisolone. The levels of the above markers were measured by Enzyme linked immunosorbent assay (ELISA) and compared with the individuals own value in the month prior to the reaction and presented as percentage increase. One month before the reaction individuals showed a varying increase in the level of different markers such as TNF-α (53%) and antibodies to Ceramide (53%), followed by to PGL-1 (51%), S100B (50%) and LAM (26%). The increase was significantly associated with clinical finding of nerve pain, tenderness and new nerve function impairment. After one month prednisolone therapy, there was a fall in the levels [TNF-α (60%), C2-Ceramide (54%), S100B (67%), PGL-1(47%) and LAM (52%)] with each marker responding differently to steroid. Reactions in leprosy are inflammatory processes wherein a rise in set of serological markers can be detected a month before the clinical onset of reaction, some of which remain elevated during their action and steroid treatment induces a variable fall in the levels, and this forms the basis for a variable individual response to steroid therapy.


An approach to trial design and analysis in the era of non-proportional hazards of the treatment effect.
Most randomized controlled trials with a time-to-event outcome are designed and analysed under the proportional hazards assumption, with a target hazard ratio for the treatment effect in mind. However, the hazards may be non-proportional. We address how to design a trial under such conditions, and how to analyse the results. We propose to extend the usual approach, a logrank test, to also include the Grambsch-Therneau test of proportional hazards. We test the resulting composite null hypothesis using a joint test for the hazard ratio and for time-dependent behaviour of the hazard ratio. We compute the power and sample size for the logrank test under proportional hazards, and from that we compute the power of the joint test. For the estimation of relevant quantities from the trial data, various models could be used; we advocate adopting a pre-specified flexible parametric survival model that supports time-dependent behaviour of the hazard ratio. We present the mathematics for calculating the power and sample size for the joint test. We illustrate the methodology in real data from two randomized trials, one in ovarian cancer and the other in treating cellulitis. We show selected estimates and their uncertainty derived from the advocated flexible parametric model. We demonstrate in a small simulation study that when a treatment effect either increases or decreases over time, the joint test can outperform the logrank test in the presence of both patterns of non-proportional hazards. Those designing and analysing trials in the era of non-proportional hazards need to acknowledge that a more complex type of treatment effect is becoming more common. Our method for the design of the trial retains the tools familiar in the standard methodology based on the logrank test, and extends it to incorporate a joint test of the null hypothesis with power against non-proportional hazards. For the analysis of trial data, we propose the use of a pre-specified flexible parametric model that can represent a time-dependent hazard ratio if one is present.


Using the one-lung method to link p38 to pro-inflammatory gene expression during overventilation in C57BL/6 and BALB/c mice.
The mechanisms of ventilator-induced lung injury (VILI), including the role of MAP kinases, are frequently studied in different mouse strains. A useful model for such studies is the isolated perfused mouse lung. As a further development we present the one-lung method that permits to continue perfusion and ventilation of the right lung after removal of the left lung. This method was used to compare the effect of high pressure ventilation (HPV) on pro-inflammatory signaling events in two widely used mouse strains (C57BL/6, BALB/c) and to further define the role of p38 in VILI. Lungs were perfused and ventilated for 30 min under control conditions before they were randomized to low (8 cm H(2)O) or high (25 cm H(2)O) pressure ventilation (HPV) for 210 min, with the left lung being removed after 180 min. In the left lung we measured the phosphorylation of p38, JNK, ERK and Akt kinase, and in the right lung gene expression and protein concentrations of Il1b, Il6, Tnf, Cxcl1, Cxcl2, and Areg. Lung mechanics and kinase activation were similar in both mouse strains. HPV increased all genes (except Tnf in BALB/c) and all mediators in both strains. The gene expression of mRNA for Il1b, Il6, Cxcl1 and Cxcl2 was higher in BALB/c mice. Backward regression of the kinase data at t = 180 min with the gene and protein expression data at t = 240 min suggested that p38 controls HPV-induced gene expression, but not protein production. This hypothesis was confirmed in experiments with the p38-kinase inhibitor SB203580. The one-lung method is useful for mechanistic studies in the lungs. While C57BL/6 show diminished pro-inflammatory responses during HPV, lung mechanics and mechanotransduction processes appear to be similar in both mouse strains. Finally, the one-lung method allowed us to link p38 to gene expression during VILI.


Temporal trends in prevention of venous thromboembolism following primary total hip or knee arthroplasty 1996-2001: findings from the Hip and Knee Registry.
The Hip and Knee Registry is an observational database comprising data on practices of US orthopedic surgeons during 1996 to 2001. We examined trends in the use of prophylaxis for venous thromboembolism (VTE) among patients who underwent primary total hip arthroplasty (THA) or total knee arthroplasty (TKA). Data on 9,327 THA and 13,846 TKA patients were submitted between 1996 and 2001 by 464 orthopedic surgeons from 319 hospitals in 42 of the United States. During 1996 to 2001, 44% of THA patients and 38% of TKA patients were male, and 93% and 92% were white, respectively. The median age of THA and TKA patients increased from 66 to 68 years and 68 to 69 years, respectively, between 1996 and 2001 (p < 0.001), while the mean length of hospital stay decreased from 4.7 to 3.7 days and 4.5 to 3.7 days, respectively (p < 0.001). Use of spinal or epidural anesthesia increased from 34 to 46% for THA and 43 to 54% for TKA patients (p < 0.001). One or more types of thromboprophylaxis were administered to 99% of patients. The following were the most common types of thromboprophylaxis: elastic stockings (61% and 58%), warfarin (56% and 53%), low-molecular-weight heparin (38% and 40%), and intermittent pneumatic compression (35% and 32%) in THA and TKA patients, respectively. Aspirin was used for thromboprophylaxis in 4% of THA and 7% of TKA patients. One or more type of in-hospital prophylaxis matching the 2001 American College of Chest Physicians (ACCP) recommendations were administered to 89% of THA and 91% of TKA patients between 1996 and 2001. During this period, in-hospital use of ACCP prophylaxis recommendations increased from 88 to 94% following THA (p < 0.001). This increase was also observed for prophylaxis administered to TKA patients, although this did not reach statistical significance. Recent trends in the management of patients undergoing THA and TKA in the United States, including shorter lengths of hospital stay and increased use of spinal/epidural anesthesia, present a challenge to orthopedic surgeons who wish to provide their patients with effective prophylaxis for VTE. Despite these challenges, nearly all surgeons participating in the Hip and Knee Registry are providing types of prophylaxis consistent with evidence-based consensus recommendations. Although there are concerns regarding increased bleeding risk due to the use of anticoagulants in patients receiving spinal/epidural anesthesia, there was a significant increase in the use of spinal/epidural anesthesia between 1996 and 2001. During this same period, the proportion of patients receiving spinal/epidural anesthesia who were also administered anticoagulants as prophylaxis increased significantly.


Variability in krill biomass links harvesting and climate warming to penguin population changes in Antarctica.
The West Antarctic Peninsula (WAP) and adjacent Scotia Sea support abundant wildlife populations, many of which were nearly extirpated by humans. This region is also among the fastest-warming areas on the planet, with 5-6 °C increases in mean winter air temperatures and associated decreases in winter sea-ice cover. These biological and physical perturbations have affected the ecosystem profoundly. One hypothesis guiding ecological interpretations of changes in top predator populations in this region, the "sea-ice hypothesis," proposes that reductions in winter sea ice have led directly to declines in "ice-loving" species by decreasing their winter habitat, while populations of "ice-avoiding" species have increased. However, 30 y of field studies and recent surveys of penguins throughout the WAP and Scotia Sea demonstrate this mechanism is not controlling penguin populations; populations of both ice-loving Adélie and ice-avoiding chinstrap penguins have declined significantly. We argue in favor of an alternative, more robust hypothesis that attributes both increases and decreases in penguin populations to changes in the abundance of their main prey, Antarctic krill. Unlike many other predators in this region, Adélie and chinstrap penguins were never directly harvested by man; thus, their population trajectories track the impacts of biological and environmental changes in this ecosystem. Linking trends in penguin abundance with trends in krill biomass explains why populations of Adélie and chinstrap penguins increased after competitors (fur seals, baleen whales, and some fishes) were nearly extirpated in the 19th to mid-20th centuries and currently are decreasing in response to climate change.


Impact of intrapeptide epitope location on CD8 T cell recognition: implications for design of overlapping peptide panels.
Antigen-specific CD8 T cells following infection or immunization are typically assessed by measuring interferon-gamma production after stimulation with overlapping peptides spanning the region of interest. The effect of epitope location within such peptides is not known but may influence recognition. To examine if peptides containing the appropriate C-terminal anchor amino acid residue would provide more sensitive detection of T cell responses. The impact was examined of epitope location within overlapping peptides on recognition of epitope-specific CD8 T cell responses. C-terminal amino acid residues were analyzed in well-defined optimal epitopes for HIV, Epstein-Barr virus, cytomegalovirus and influenza and in peptide-binding motifs. Recognition of known epitopes within longer synthesized peptides by peripheral blood mononuclear cells or CD8 T cell lines was tested using interferon-gamma Elispot at various peptide concentrations. Only 9 of 20 amino acids served as the C-terminal anchor position in 96% of described optimal epitopes and in 95% of peptide-binding motifs. A CD8 T cell response to an epitope within a longer peptide is best detected when the epitope is situated at the C-terminal end of the longer peptide, both when using peptides designed to include the optimal epitope at every possible position and when comparing responses towards optimal epitopes and corresponding overlapping peptides in a larger group of subjects. When using overlapping peptides to screen for CD8 T cell responses, more sensitive detection will be achieved using known C-terminal anchor amino acid residues at the C-terminus.


[Quantitative magnetic resonance tomography and the severity of deficits in dementia of the Alzheimer type].
The aim of the present study was to investigate the severity of dementia of the Alzheimer type (DAT) with respect to morphological changes revealed by quantitative magnetic resonance imaging (MRI). Seventeen patients with DAT (NINCDS-ADRDA guidelines) and 10 healthy elderly controls were included. The severity of dementia was evaluated on the Mini Mental State Examination (MMSE), the Global Deterioration Scale (GDS) and the Brief Cognitive Rating Scale (BCRS). Three-dimensional MRI sequences were acquired using a 1.5-T Siemens Magnetom. Whole-brain volume and the volume of the amygdala-hippocampus complex (AHC) were assessed using the newly developed software NMRWin. This software provides a semi-automated measure of the whole brain volume, while measurement of substructures requires manual guidance. In addition, the ventricle-brain ratio (VBR) was assessed. All morphometric parameters differed significantly between the two groups. AHC volumes discriminated best between them with only a small overlap. These findings were confirmed when only the data of the mildly demented patients were included in the analysis. The degree of AHC atrophy exceeded that of generalized cerebral atrophy. The severity of dementia as indicated by the MMSE, GDS and BCRS scores was correlated significantly with the volumes of the AHC bilaterally, but not with whole-brain volume or with VBR. These results underline the importance of the mesial temporal substructures in the etiology and progression of DAT and indicate that the volume of the AHC can be monitored by MRI and may be used to follow up the disease process.


Progesterone as a mediator of gonadotrophin action in the corpus luteum: beyond steroidogenesis.
Studies using newer, potent GnRH antagonists and pure gonadotrophins have clarified the importance of: (i) the strength-duration of the midcycle surge of pituitary gonadotrophins (LH, FSH) in follicle rupture and conversion to the corpus luteum; (ii) the continued requirement for pituitary LH throughout development and the functional lifespan of the primate corpus luteum in the menstrual cycle; and (iii) the exponential secretion of chorionic gonadotrophin (CG) by the developing placenta to extend the functional lifespan of the primate corpus luteum in early pregnancy. Although studies continue to increase current understanding of the cellular and molecular actions of LH/CG to stimulate luteal steroidogenesis, knowledge of the processes whereby these gonadotrophins promote the development and maintenance of the functional corpus luteum remains limited. This review summarizes evidence that the primate ovulatory follicle and corpus luteum is a target for the primary steroid produced by luteinizing/luteal tissue (i.e. progesterone). With evidence for dynamic expression of genomic progesterone receptors (PRA/B), and possibly other progesterone-receptor systems, recent studies addressed the hypothesis that progesterone is a critical 'local luteotrophin' that promotes luteal development and sustains luteal structure-function during the menstrual cycle and early pregnancy. Specific progesterone actions to regulate tissue remodelling (via protease expression), health (anti-apoptotic effects) and sensitivity to other local factors (e.g. via estrogen receptor expression) are discussed. The collective data suggest that there are gonadotrophin-stimulated, progesterone-dependent processes that promote luteotrophic and suppress luteolytic pathways in the primate corpus luteum. However, further studies are needed to verify their role in normal ovarian function and relevance to possible ovarian defects in natural and assisted reproduction technique-related cycles.


Microtiter virus isolation and enzyme immunoassays for detection of bovine viral diarrhea virus in cattle serum.
Cattle immunotolerant to and persistently infected (PI) with bovine viral diarrhea (BVD) virus (BVDV) constitute the mechanism by which BVDV persists in cattle herds. Two procedures for using serum to detect PI cattle were developed and evaluated. BVDV was found to remain viable for 7 days in serum samples stored at room temperature. The tests use cell culture virus isolation (VI) in 96-well microtiter plates, followed by immunostaining of cell monolayers with monoclonal antibodies. One technique, the immunoperoxidase monolayer assay (IPMA), forms a red intracellular precipitate while the other, the monolayer enzyme-linked immunosorbent assay (M-ELISA) produces a yellow color in solution. The optimal incubation period for microtiter VI was determined to be 4 days. Optimal IPMA staining was obtained by fixing cell monolayers with 20 to 30% acetone, whereas a simple dry-rehydrate-dry cycle provided optimal M-ELISA staining. The M-ELISA and IPMA had the same sensitivities and specificities, but the M-ELISA was a more rapid procedure and use of a spectrophotometer for reading samples allowed for greater objectivity. When compared to standard VI with routine samples submitted for the diagnosis of BVD, M-ELISA and IPMA had a relative sensitivity of 85% and a relatively specificity of 100%. When only samples from cattle suspected of being PI were considered, these two parameters were 100% for both IPMA and M-ELISA. The two procedures, especially the M-ELISA, are suitable for whole-herd testing to identify PI cattle. The appeal of these tests is derived from the convenience of using serum as a diagnostic sample and the ability to rapidly screen large numbers of samples at low cost.


Detecting intracellular thiol redox state in leukaemia and heterogeneous immune cell populations: An optimised protocol for digital flow cytometers.
Flow cytometric methods for detecting and quantifying reduced intracellular thiol content using fluorescein-5-maleimide (F5M) in viable eukaryotic cells date back to 1983 (Durand and Olive [1]). There has been little development in these methodologies since that time, a period that has witnessed huge technological advances, particularly with the emergence of digital multi-parameter flow cytometric systems. Concurrent advancement in our understanding of redox regulation within eukaryotic cellular systems has also followed, whereby it is now accepted that cysteine thiols partake in redox reactions, which regulate protein activity and function (Groitl and Jakob (2014), Won et al. (2012)). Moreover, we are at the dawn of a new era in redox biology whereby the importance of 'reductive stress' in eukaryotic cellular systems is gathering momentum (Wadley et al. (2018) [4]). It is therefore critical that methods be continually advanced to better understand these concepts in more detail at the cellular level. Flow cytometry is a powerful technique that may be used for this purpose. Henceforth we have rejuvenated these methods to address modern scientific questions. In this paper, essential detail is provided on: •The adaption of a protocol initially described by Durand and Olive [1] for use with modern digital flow cytometer configurations. Here we provide optimal conditions for labelling intracellular thiols with F5M for detection using digital flow cytometers. Our modifications avoid the use of methanol fixation thus preserving cell viability in single cell suspension cultures.•Demonstration that flow cytometry can detect the gain and loss of reduced intracellular thiols in cells exposed to physiological doses of hydrogen peroxide mediated by glucose oxidase (Hole et al. (2013) [5]).•Validation of F5M protein labelling by coupling method to confocal microscopy and downstream proteomics, thus permitting a powerful experimental platform for potential use with next generation flow cytometry e.g. CyTOF (Lin and Maecker (2018) [6]).


Dose intensity in small cell lung cancer.
Evidence from preclinical models and from clinical trials describing the importance of dose intensity in securing a better treatment outcome is reviewed. Recent randomized trials have shown statistically significant survival benefits with higher-dose, accelerated chemotherapy regimens with and without granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. The novel use of peripheral blood progenitor cells contained in whole blood autotransfusions, which allow a marked increase in dose intensity of an ifosfamide/carboplatin/etoposide regimen, could provide a much easier method of delivering dose-intensive chemotherapy than previously available.


Ecology of arthritis.
Osteoarthritis (OA) is a widespread degenerative disease of skeletal joints and is often associated with senescence in vertebrates. OA commonly results from excessive or abnormal mechanical loading of weight-bearing joints ('wear-and-tear'), arising from heavy long-term use or specific injuries; yet, in the absence of injury, the aetiology of OA remains obscure. We show that poor nutritional conditions experienced by moose (Alces alces) early in life are linked to greater prevalence of OA during senescence as well as reduced life expectancy. Moreover, we also found a negative relationship between kill rate by wolves (Canis lupus) and prevalence of OA, suggesting a potential connection between senescence of prey and the population ecology of predator-prey systems. This association between OA and early malnutrition also provides a basis for explaining the observation in anthropology that OA became more prevalent in native Americans as their diet become poorer - the result of relying more on corn and agriculture and less on hunting and gathering.


Partitioning heritability by functional annotation using genome-wide association summary statistics.
Recent work has demonstrated that some functional categories of the genome contribute disproportionately to the heritability of complex diseases. Here we analyze a broad set of functional elements, including cell type-specific elements, to estimate their polygenic contributions to heritability in genome-wide association studies (GWAS) of 17 complex diseases and traits with an average sample size of 73,599. To enable this analysis, we introduce a new method, stratified LD score regression, for partitioning heritability from GWAS summary statistics while accounting for linked markers. This new method is computationally tractable at very large sample sizes and leverages genome-wide information. Our findings include a large enrichment of heritability in conserved regions across many traits, a very large immunological disease-specific enrichment of heritability in FANTOM5 enhancers and many cell type-specific enrichments, including significant enrichment of central nervous system cell types in the heritability of body mass index, age at menarche, educational attainment and smoking behavior.


Toward a Bioecological Model of School Engagement: A Biometric Analysis of Gene and Environmental Factors.
School disengagement is associated with poor academic achievement, dropout, and risk behaviors such as truancy, delinquency, and substance use. Despite empirical research identifying risk correlates of school disengagement across the ecology, it is unclear from which domain these correlates arise. To redress this issue, the current study used intraclass correlation and DeFries-Fulker analyses to longitudinally decompose variance in three domains of engagement (academic, behavioral, and emotional) using data from the National Longitudinal Study of Adolescent Health. Findings suggest that nonshared environmental factors (that is, environmental contexts and experiences that are unique to each sibling) account for approximately half of the variance in indicators of school disengagement when controlling for genetic influences, and that this variance increases as adolescents grow older and rely less on their immediate family. The present study contributes new evidence on the biosocial underpinnings of school engagement and highlights the importance of interventions targeting factors in the nonshared environment.


Radiocarbon constraints on the extent and evolution of the South Pacific glacial carbon pool.
During the last deglaciation, the opposing patterns of atmospheric CO2 and radiocarbon activities (Δ(14)C) suggest the release of (14)C-depleted CO2 from old carbon reservoirs. Although evidences point to the deep Pacific as a major reservoir of this (14)C-depleted carbon, its extent and evolution still need to be constrained. Here we use sediment cores retrieved along a South Pacific transect to reconstruct the spatio-temporal evolution of Δ(14)C over the last 30,000 years. In ∼2,500-3,600 m water depth, we find (14)C-depleted deep waters with a maximum glacial offset to atmospheric (14)C (ΔΔ(14)C=-1,000‰). Using a box model, we test the hypothesis that these low values might have been caused by an interaction of aging and hydrothermal CO2 influx. We observe a rejuvenation of circumpolar deep waters synchronous and potentially contributing to the initial deglacial rise in atmospheric CO2. These findings constrain parts of the glacial carbon pool to the deep South Pacific.


Synergizing radiation therapy and immunotherapy for curing incurable cancers. Opportunities and challenges.
The combination of radiation therapy and immunotherapy holds particular promise as a strategy for cancer therapeutics. Evidence suggests that immunotherapy is most beneficial alone when employed early in the disease process or in combination with standard therapies (eg, radiation) later in the disease process. Indeed, radiation may act synergistically with immunotherapy to enhance immune responses, inhibit immunosuppression, and/or alter the phenotype of tumor cells, thus rendering them more susceptible to immune-mediated killing. As monotherapies, both immunotherapy and radiation may be insufficient to eliminate tumor masses. However, following immunization with a cancer vaccine, the destruction of even a small percentage of tumor cells by radiation could result in crosspriming and presentation of tumor antigens to the immune system, thereby potentiating antitumor responses. Learning how to exploit radiation-induced changes to tumor-cell antigens, and how to induce effective immune responses to these cumulatively immunogenic stimuli, is an exciting frontier in cancer therapy research. This review examines mechanisms by which many forms of radiation therapy can induce or augment antitumor immune responses as well as preclinical systems demonstrating that immunotherapy can be effectively combined with radiation therapy. Finally, we review current clinical trials where standard-of-care radiation therapy is being combined with immunotherapy.


Hyperglycemia inhibits insulin activation of Akt/protein kinase B but not phosphatidylinositol 3-kinase in rat skeletal muscle.
Sustained hyperglycemia impairs insulin-stimulated glucose utilization in the skeletal muscle of both humans and experimental animals--a phenomenon referred to clinically as glucose toxicity. To study how this occurs, a model was developed in which hyperglycemia produces insulin resistance in vitro. Rat extensor digitorum longus muscles were preincubated for 4 h in Krebs-Henseleit solution containing glucose or glucose + insulin at various concentrations, after which insulin action was studied. Preincubation with 25 mmol/l glucose + insulin (10 mU/ml) led to a 70% decrease in the ability of insulin (10 mU/ml) to stimulate glucose incorporation into glycogen and a 30% decrease in 2-deoxyglucose (2-DG) uptake, compared with muscles incubated with 0 mmol/l glucose. Glucose incorporation into lipid and its oxidation to CO2 were marginally diminished, if at all. The alterations of glycogen synthesis and 2-DG uptake were first evident after 1 h and were maximal after 2 h of preincubation; they were not observed in muscles preincubated with 25 mmol/l glucose + insulin for 5 min. Preincubation for 4 h with 25 mmol/l glucose in the absence of insulin produced a similar although somewhat smaller decrease in insulin-stimulated glycogen synthesis; however, it did not alter 2-DG uptake, glucose oxidation to CO2, or incorporation into lipids. Studies of insulin signaling in the latter muscles revealed that activation of Akt/protein kinase B (PKB) was diminished by 60%, compared with that of muscles preincubated in a glucose-free medium; whereas activation of phosphatidylinositol (PI) 3-kinase, an upstream regulator of Akt/PKB in the insulin-signaling cascade, and of mitogen-activated protein (MAP) kinase, a parallel signal, was unaffected. Immunoblots demonstrated that this was not due to a change in Akt/PKB abundance. The results indicate that hyperglycemia-induced insulin resistance can be studied in rat skeletal muscle in vitro. They suggest that impairment of insulin action in these muscles is related to inhibition of Akt/PKB by events that do not affect PI 3-kinase.


The transcriptional modulator Ifrd1 controls PGC-1α expression under short-term adrenergic stimulation in brown adipocytes.
Sympathetic tone activates the function of classical brown adipocytes, which constitutively exist in the brown adipose tissue (BAT), and inducible brown adipocytes (so-called beige adipocytes), which sporadically reside within the white adipose tissue (WAT). Here we identified the transcriptional modulator interferon-related developmental regulator 1 (Ifrd1) as a negative regulator of thermogenic and mitochondrial gene expression in brown adipocytes. Ifrd1 expression was markedly induced by cold exposure and administration of CL-316243 (a β3 adrenergic agonist) in interscapular brown adipose and inguinal subcutaneous WATs, but not in epididymal visceral WAT, in vivo. Adrenergic stimulation also induced Ifrd1 expression in brown adipocytes in a cAMP responsive element binding protein-dependent manner in vitro. CL-316243 injection markedly elevated thermogenic and mitochondrial gene expression, including peroxisome proliferator-activated receptor γ coactivator 1α (Pgc1a) in the subcutaneous WAT of Ifrd1 knockout mice compared with gene expression in wild-type mice. Pgc1a promoter activity enhanced by the transcription factor specificity protein 1 (Sp1) was markedly repressed by co-introduction of Ifrd1 in brown adipocytes, whereas the repression was markedly prevented by the addition of trichostatin A, a histone deacetylase inhibitor. Moreover, adrenergic stimulation induced complex formation between Ifrd1, Sp1 and mSIN3B, which is a component of the SIN complex containing histone deacetylase, in brown adipocytes. These findings, therefore, suggest that Ifrd1 could be a pivotal negative regulator of sympathetic regulation of thermogenic and mitochondrial gene expression in brown adipocytes by interacting with Sp1 and the mSIN3 complex.


Somatosensory amplification, health anxiety, and alexithymia in generalized anxiety disorder.
The aim of this study was to study somatosensory amplification, health anxiety (hypochondriasis), and alexithymia among patients with generalized anxiety disorder (GAD) and to evaluate the association of these variables with the severity of GAD. Cross-sectional design was employed, and patients were recruited from the outpatient clinic of the psychiatry department of a multispecialty tertiary care medical institute in North India. The patients who were clinically diagnosed to have GAD by the two independent qualified psychiatrists were screened with Mini International Neuropsychiatry Interview to confirm the diagnosis. Forty patients with GAD meeting the inclusion criteria were assessed with GAD-7 scale, somatosensory amplification scale (SSAS), the Whiteley Index (WI) and Toronto alexithymia scale - 20 Hindi version (TAS-H-20). The mean scores of patients with GAD on SSAS, WI, TAS-H-20, and GAD-7 scale were 25.70 (SD-5.84), 7.75 (SD-3.30), 59.77 (SD- 8.63), and 13.37 (SD- 3.58), respectively. Half of the patients with GAD had significant health anxiety as defined by WI score of >7. Around 40% of GAD patients were alexithymic as defined with TAS-H-20 scores of >60. SSAS, WI, TAS-H-20 had a positive correlation with the severity of GAD as measured with GAD-7 scale. GAD patients have significant somatosensory amplification, health anxiety (hypochondriasis), and alexithymia. Accordingly, there is a need to develop effective psychological interventions focused on these factors in GAD.


Short Communication: Comparative Evaluation of Coformulated Injectable Combination Antiretroviral Therapy Regimens in Simian Immunodeficiency Virus-Infected Rhesus Macaques.
The use of nonhuman primate (NHP) models to study persistent residual virus and viral eradication strategies in combination antiretroviral therapy (cART)-treated individuals requires regimens that effectively suppress SIV replication to clinically relevant levels in macaques. We developed and evaluated two novel cART regimens in SIVmac239-infected rhesus macaques: (1) a "triple regimen" containing the nucleo(s/t)ide reverse transcriptase inhibitors emtricitabine (FTC) and tenofovir disoproxil fumarate [TDF, prodrug of tenofovir (TFV, PMPA)] with the integrase strand transfer inhibitor dolutegravir (DTG) (n = 3), or (2) a "quad regimen" containing the same three drugs plus the protease inhibitor darunavir (DRV) (n = 3), with each regimen coformulated for convenient administration by a single daily subcutaneous injection. Plasma drug concentrations were consistent across animals within the triple and quad regimen-treated groups, although DTG levels were lower in the quad regimen animals. Time to achieve plasma viral loads stably <30 viral RNA copies/ml ranged from 12 to 20 weeks of treatment between animals, and viral loads <30 viral RNA copies/ml plasma were maintained through 40 weeks of follow-up on cART. Notably, although we show virologic suppression and development of viral resistance in a separate cohort of SIV-infected animals treated with oral DRV monotherapy, the addition of DRV in the quad regimen did not confer an apparent virologic benefit during early treatment, hence the quad regimen-treated animals were switched to the triple regimen after 4 weeks. This coformulated triple cART regimen can be safely, conveniently, and sustainably administered to durably suppress SIV replication to clinically relevant levels in rhesus macaques.


Toward a new history and geography of human genes informed by ancient DNA.
Genetic information contains a record of the history of our species, and technological advances have transformed our ability to access this record. Many studies have used genome-wide data from populations today to learn about the peopling of the globe and subsequent adaptation to local conditions. Implicit in this research is the assumption that the geographic locations of people today are informative about the geographic locations of their ancestors in the distant past. However, it is now clear that long-range migration, admixture, and population replacement subsequent to the initial out-of-Africa expansion have altered the genetic structure of most of the world's human populations. In light of this we argue that it is time to critically reevaluate current models of the peopling of the globe, as well as the importance of natural selection in determining the geographic distribution of phenotypes. We specifically highlight the transformative potential of ancient DNA. By accessing the genetic make-up of populations living at archaeologically known times and places, ancient DNA makes it possible to directly track migrations and responses to natural selection.


High Resolution pH Measurements Using a Lab-on-Chip Sensor in Surface Waters of Northwest European Shelf Seas.
Increasing atmospheric CO₂ concentrations are resulting in a reduction in seawater pH, with potential detrimental consequences for marine organisms. Improved efforts are required to monitor the anthropogenically driven pH decrease in the context of natural pH variations. We present here a high resolution surface water pH data set obtained in summer 2011 in North West European Shelf Seas. The aim of our paper is to demonstrate the successful deployment of the pH sensor, and discuss the carbonate chemistry dynamics of surface waters of Northwest European Shelf Seas using pH and ancillary data. The pH measurements were undertaken using spectrophotometry with a Lab-on-Chip pH sensor connected to the underway seawater supply of the ship. The main processes controlling the pH distribution along the ship's transect, and their relative importance, were determined using a statistical approach. The pH sensor allowed 10 measurements h-1 with a precision of 0.001 pH units and a good agreement with pH calculated from a pair of discretely sampled carbonate variables dissolved inorganic carbon (DIC), total alkalinity (TA) and partial pressure of CO₂ (pCO₂) (e.g., pHDICpCO2). For this summer cruise, the biological activity formed the main control on the pH distribution along the cruise transect. This study highlights the importance of high quality and high resolution pH measurements for the assessment of carbonate chemistry dynamics in marine waters.


Transducin β-like protein 1 recruits nuclear factor κB to the target gene promoter for transcriptional activation.
Nuclear factor κB (NF-κB) signaling controls a wide range of cellular functions such as tumor progression and invasion by inducing gene expression. Upon stimulation, NF-κB is translocated to the nucleus and binds to its target gene promoters to activate transcription by recruiting transcription coactivators. Although significant progress has been made in understanding NF-κB-mediated transactivation, little is known about how NF-κB is recruited to its target gene promoters. Here, we report that transducin β-like protein 1 (TBL1) controls the expression of NF-κB target genes by directly binding with NF-κB and facilitating its recruitment to target gene promoters. Tumor necrosis factor alpha stimulation triggered the formation of an NF-κB and TBL1 complex and subsequent target gene promoter binding. Knockdown of TBL1 impaired the recruitment of NF-κB to its target gene promoters. Interestingly, analysis of the Oncomine database revealed that TBL1 mRNA levels were significantly higher in invasive breast cancer tissues than in breast adenocarcinoma tissue. Consistently, TBL1 knockdown significantly reduced the invasive potential of breast cancer cells by inhibiting NF-κB. Our results reveal a new mechanism for the regulation of NF-κB activation, with important implications for the development of novel strategies for cancer therapy by targeting NF-κB.


Societal reintegration after liver transplantation: findings in alcohol-related and non-alcohol-related transplant recipients.
To compare the degree of societal reintegration between alcohol-related and non-alcohol-related liver transplant recipients. Orthotopic liver transplantation (OLTX) is the treatment of choice for end-stage liver disease of various etiologies. Returning patients to society to lead active and productive lives is a key goal of OLTX. A questionnaire assessing societal reintegration was administered by phone to 84 alcoholic liver disease (ALD) OLTX recipients (ALDs) and 68 non-ALD OLTX recipients having undergone OLTX at a single-center urban not-for-profit teaching hospital. Sixty-eight non-ALD OLTX recipients, serving as the control group (Controls), were matched to the ALDs by age, sex, and length of follow-up. Participation levels were assessed in the following areas: employment, homemaking, academic study, support of others through financial and/or care-giving efforts, and involvement in social or community groups and activities. Seventy-nine percent of ALDs and 81% of Controls were male. Median age was 53 years for ALDs and 54 years for Controls. Median length of follow-up for both groups separately was 52 months. No significant differences were noted between ALDs and Controls in the proportion of employed individuals, homemakers, students, and supporters of others. Controls were significantly more likely than ALDs to be involved in structured social activities and routine volunteer work. Alcohol-related and non-alcohol-related OLTX recipients appear to return to society to lead similarly active and productive lives. ALD OLTX recipients appear less likely to be involved in structured social activities and routine volunteer work than non-ALD OLTX recipients.


Consumption of Whole Grains, Refined Cereals, and Legumes and Its Association With Colorectal Cancer Among Jordanians.
Background The role of whole grains, refined cereals, and legumes in preventing or initiating colorectal cancer (CRC) is still uncertain. The aim of this study is to examine the possible association between the consumption of whole grains, refined cereals, and legumes and the risk of developing CRC among Jordanian population. Methods A validated food frequency questionnaire was used to collect dietary data with regard to intake of whole grains, refined cereals, and legumes. A total of 220 diagnosed CRC participants and 281 CRC-free control participants matched by age, gender, occupation, and marital status were recruited. Logistic regression was used to estimate the odds of developing CRC in relation to the consumption of different types of whole grains, refined cereals, and legumes. Results The odds ratio (OR) for developing CRC among cases consumed refined wheat bread at all meals was 3.1 compared with controls (95% CI: 1.2-7.9, P-Trend = 0.001); whereas the OR associated with whole wheat bread was 0.44 (95% CI: 0.22-0.92, P-Trend = 0.001). The statistical evaluation for daily consumption of rice suggested a direct association with the risk of developing CRC, OR = 3.0 (95% CI: 0.27-33.4, P-Trend = 0.020). Weekly consumption of macaroni was associated with CRC with OR of 2.4 (95% CI: 1.1-5.3, P-Trend = 0.001). The consumption of corn, bulgur, lentils, and peas suggested a protective trend, although the trend was not statistically significant. Conclusion This study provides additional indicators of the protective role of whole grains and suggests a direct association between consumption of refined grains and higher possibility for developing CRC.


Rehabilitation service utilization and determinants among people with an intellectual disability: preliminary findings in Taiwan.
Improving rehabilitation services for people with intellectual disabilities (ID) remains an ongoing challenge in the public health system. The purpose of this article was to investigate the types of rehabilitation services used by people with ID and determine what factors predict resource utilization in Taiwan. Samples of 957 people with ID were recruited from the Taiwan National Disability Register in a cross-sectional study in 2001. The findings indicated that 24.5% of individuals with ID had received rehabilitation services in the past 7 months. The main types of services used were speech and communication therapy (50%), psychotherapy (32.1%), occupational therapy (30.3%) and physiotherapy (25.2%). Stepwise logistic regression was carried out for the utilization of rehabilitation services (yes/no). The model revealed that the following factors: (i) Major Illness Card holder, (ii) time spent in medical care, (iii) having a family physician, (iv) having illnesses, (v) age of ID individual, and (vi) gender of the main carer, were all significantly associated with the utilization of rehabilitation services. We should reorient the healthcare system to respond adequately to the health needs of rehabilitation service users and its determinants, and further research should focus on the effectiveness and efficiency of rehabilitation for people with ID in Taiwan.


Risk Factor Detection as a Metric of STARHS Performance for HIV Incidence Surveillance Among Female Sex Workers in Kigali, Rwanda.
The epidemiologic utility of STARHS hinges not only on producing accurate estimates of HIV incidence, but also on identifying risk factors for recent HIV infection. As part of an HIV seroincidence study, 800 Rwandan female sex workers (FSW) were HIV tested, with those testing positive further tested by BED-CEIA (BED) and AxSYM Avidity Index (Ax-AI) assays. A sample of HIV-negative (N=397) FSW were followed prospectively for HIV seroconversion. We compared estimates of risk factors for: 1) prevalent HIV infection; 2) recently acquired HIV infection (RI) based on three different STARHS classifications (BED alone, Ax-AI alone, BED/Ax-AI combined); and 3) prospectively observed seroconversion. There was mixed agreement in risk factors between methods. HSV-2 coinfection and recent STI treatment were associated with both prevalent HIV infection and all three measures of recent infection. A number of risk factors were associated only with prevalent infection, including widowhood, history of forced sex, regular alcohol consumption, prior imprisonment, and current breastfeeding. Number of sex partners in the last 3 months was associated with recent infection based on BED/Ax-AI combined, but not other STARHS-based recent infection outcomes or prevalent infection. Risk factor estimates for prospectively observed seroconversion differed in magnitude and direction from those for recent infection via STARHS. Differences in risk factor estimates by each method could reflect true differences in risk factors between the prevalent, recently, or newly infected populations, the effect of study interventions (among those followed prospectively), or assay misclassification. Similar investigations in other populations/settings are needed to further establish the epidemiologic utility of STARHS for identifying risk factors, in addition to incidence rate estimation.


Role of dosage-sensitive sex reversal, adrenal hypoplasia congenita, critical region on the X chromosome, gene 1 in protein kinase A- and protein kinase C-mediated regulation of the steroidogenic acute regulatory protein expression in mouse Leydig tumor cells: mechanism of action.
Dosage-sensitive sex reversal, adrenal hypoplasia congenita, critical region on the X chromosome, gene 1 (DAX-1) is an orphan nuclear receptor that has been demonstrated to be instrumental to the expression of the steroidogenic acute regulatory (StAR) protein that regulates steroid biosynthesis in steroidogenic cells. However, its mechanism of action remains obscure. The present investigation was aimed at exploring the molecular involvement of DAX-1 in protein kinase A (PKA)- and protein kinase C (PKC)-mediated regulation of StAR expression and its concomitant impact on steroid synthesis using MA-10 mouse Leydig tumor cells. We demonstrate that activation of the PKA and PKC pathways, by a cAMP analog dibutyryl (Bu)2cAMP [(Bu)2cAMP] and phorbol 12-myristate 13-acetate (PMA), respectively, markedly decreased DAX-1 expression, an event that was inversely correlated with StAR protein, StAR mRNA, and progesterone levels. Notably, the suppression of DAX-1 requires de novo transcription and translation, suggesting that the effect of DAX-1 in regulating StAR expression is dynamic. Chromatin immunoprecipitation studies revealed the association of DAX-1 with the proximal but not the distal region of the StAR promoter, and both (Bu)2cAMP and PMA decreased in vivo DAX-1-DNA interactions. EMSA and reporter gene analyses demonstrated the functional integrity of this interaction by showing that DAX-1 binds to a DNA hairpin at position -44/-20 bp of the mouse StAR promoter and that the binding of DAX-1 to this region decreases progesterone synthesis by impairing transcription of the StAR gene. In support of this, targeted silencing of endogenous DAX-1 elevated basal, (Bu)2cAMP-, and PMA-stimulated StAR expression and progesterone synthesis. Transrepression of the StAR gene by DAX-1 was tightly associated with expression of the nuclear receptors Nur77 and steroidogenic factor-1, demonstrating these factors negatively modulate the steroidogenic response. These findings provide insight into the molecular events by which DAX-1 influences the PKA and PKC signaling pathways involved in the regulation of the StAR protein and steroidogenesis in mouse Leydig tumor cells.


Identifying Crohn's disease signal from variome analysis.
After years of concentrated research efforts, the exact cause of Crohn's disease (CD) remains unknown. Its accurate diagnosis, however, helps in management and preventing the onset of disease. Genome-wide association studies have identified 241 CD loci, but these carry small log odds ratios and are thus diagnostically uninformative. Here, we describe a machine learning method-AVA,Dx (Analysis of Variation for Association with Disease)-that uses exonic variants from whole exome or genome sequencing data to extract CD signal and predict CD status. Using the person-specific coding variation in genes from a panel of only 111 individuals, we built disease-prediction models informative of previously undiscovered disease genes. By additionally accounting for batch effects, we were able to accurately predict CD status for thousands of previously unseen individuals from other panels. AVA,Dx highlighted known CD genes including NOD2 and new potential CD genes. AVA,Dx identified 16% (at strict cutoff) of CD patients at 99% precision and 58% of the patients (at default cutoff) with 82% precision in over 3000 individuals from separately sequenced panels. Larger training panels and additional features, including other types of genetic variants and environmental factors, e.g., human-associated microbiota, may improve model performance. However, the results presented here already position AVA,Dx as both an effective method for revealing pathogenesis pathways and as a CD risk analysis tool, which can improve clinical diagnostic time and accuracy. Links to the AVA,Dx Docker image and the BitBucket source code are at https://bromberglab.org/project/avadx/ .


Effect of varying hamstring tension on anterior cruciate ligament strain during in vitro impulsive knee flexion and compression loading.
The hamstring muscles are well positioned to limit both anterior tibial translation and anterior cruciate ligament strain during the knee flexion phase of a jump landing. We hypothesized that systematically increasing or decreasing hamstring tension during the knee flexion phase of a simulated jump landing would significantly affect peak relative strain in the anterior cruciate ligament. Ten cadaveric knees from four male and six female donors (mean age [and standard deviation] at the time of death, 60.3 +/- 23.6 years) were mounted in a custom fixture to initially position the specimen in 25 degrees of knee flexion and simulate axial impulsive loading averaging 1700 N to cause an increase in knee flexion. Quadriceps, hamstring, and gastrocnemius muscle forces were simulated with use of pretensioned linear springs, with the tension in the hamstrings arranged to be increased, held constant, decreased, at "baseline," or absent during knee flexion. Impulsive loading applied along the tibia and femur was monitored with use of triaxial load transducers, while uniaxial load cells monitored quadriceps and medial and lateral hamstring forces. Relative strain in the anterior cruciate ligament was measured with use of a differential variable reluctance transducer, and tibiofemoral kinematics were measured optoelectronically. For each specimen, anterior cruciate ligament strains were recorded over eighty impact trials: ten preconditioning trials, ten "baseline" trials involving decreasing hamstring tension performed before and after three sets of ten trials conducted with increasing hamstring tension, constant hamstring tension, or no hamstring tension. Peak relative strains in the anterior cruciate ligament were normalized for comparison across specimens. Increasing hamstring force during the knee flexion landing phase decreased the peak relative strain in the anterior cruciate ligament by >70% compared with the baseline condition (p = 0.005). Neither a constant hamstring muscle force nor the absence of a hamstring force significantly changed the peak strain in the anterior cruciate ligament relative to the baseline condition. Increasing hamstring muscle force during the knee flexion phase of a simulated jump landing significantly reduces the peak relative strain in the anterior cruciate ligament in vitro.


Functionalized-quantum-dot-liposome hybrids as multimodal nanoparticles for cancer.
Functionalized-quantum-dot-liposome (f-QD-L) hybrid nanoparticles are engineered by encapsulating poly(ethylene glycol)-coated QD in the internal aqueous phase of different lipid bilayer vesicles. f-QD-L maintain the QD fluorescence characteristics as confirmed by fluorescence spectroscopy, agarose gel electrophoresis, and confocal laser scanning microscopy. Cationic f-QD-L hybrids lead to dramatic improvements in cellular binding and internalization in tumor-cell monolayer cultures. Deeper penetration into three-dimensional multicellular spheroids is obtained for f-QD-L by modifying the lipid bilayer characteristics of the hybrid system. f-QD-L are injected intratumorally into solid tumor models leading to extensive fluorescent staining of tumor cells compared to injections of the f-QD alone. f-QD-L hybrid nanoparticles constitute a versatile tool for very efficient labeling of cells ex vivo and in vivo, particularly when long-term imaging and tracking of cells is sought. Moreover, f-QD-L offer many opportunities for the development of combinatory therapeutic and imaging (theranostic) modalities by incorporating both drug molecules and QD within the different compartments of a single vesicle.


Improvement of surgical wound classification following a targeted training program at a children's hospital.
Inaccurate assignment of surgical wound class (SWC) remains a challenge in perioperative documentation. The purpose of our intervention was to increase the accuracy of SWC through a targeted training program directed toward pediatric surgeons and nurses. A retrospective electronic medical record (EMR) chart review of 400 operations was performed according to NSQIP criteria during specified periods in 2014 and 2017, assessing SWC errors before and after a training program and posting of reference materials in operating rooms at a 165-bed children's hospital. After each operation, nurses confirmed SWC with the surgeon before recording the value in the EMR. Differences in proportions of misclassified SWC were evaluated with a chi-square test. Following the educational program, misclassified SWC improved from 70/200 (35.0%) to 18/200 (9.0%), p < 0.001. Misclassified SWC for appendectomies improved from 46/95 (48.4%) to 12/108 (11.1%), p < 0.001. Accurate SWC assignment in the EMR was improved by an educational program and posting of materials to aid assignment, as well as enhanced communication between surgeons and nurses at the conclusion of each operation. We present the first known attempt to list all pediatric surgery procedures according to SWC. Accurate SWC allows stratification of risks and more effective targeted interventions. Level III.


Regulation of hexokinase II activity and expression in human muscle by moderate exercise.
A single bout of exercise increases the rate of insulin-stimulated glucose uptake and metabolism in skeletal muscle. Exercise also increases insulin-stimulated glucose 6-phosphate in skeletal muscle, suggesting that exercise increases hexokinase activity. Within 3 h, exercise increases hexokinase II (HK II) mRNA and activity in skeletal muscle from rats. It is not known, however, if a single bout of moderate-intensity exercise increases HK II expression in humans. The present study was undertaken to answer this question. Six subjects had percutaneous biopsies of the vastus lateralis muscle before and 3 h after a single 3-h session of moderate-intensity aerobic (60% of maximal oxygen consumption) exercise. Glycogen synthase, HK I, and HK II activities as well as HK I and HK II mRNA content were determined from the muscle biopsy specimens. The fractional velocity of glycogen synthase was increased by 446 +/- 84% after exercise (P < 0.005). Hexokinase II activity in the soluble fraction of the homogenates increased from 1.2 +/- 0.4 to 4.5 +/- 1.6 pmol.min-1.microgram-1 (P < 0.05) but was unchanged in the particulate fraction (4.3 +/- 1.3 vs. 5.3 +/- 1.5). HK I activity in neither the soluble nor particulate fraction changed after exercise. Relative to a 28S rRNA control signal, HK II mRNA increased from 0.091 +/- 0.02 to 0.195 +/- 0.037 (P < 0.05), whereas HK I mRNA was unchanged (0.414 +/- 0.061 vs. 0.498 +/- 0.134, P < 0.20). The increase in HK II activity after moderate exercise in healthy subjects could be one factor responsible for the enhanced rate of insulin-stimulated glucose uptake seen after exercise.


Visuospatial functioning is associated with sleep disturbance and hallucinations in nondemented patients with Parkinson's disease.
Introduction: Cognitive impairment is a common symptom of Parkinson's disease (PD) associated with reduced quality of life and a more severe disease state. Previous research has shown an association between visuospatial dysfunction and worse disease course; however, it is not clear whether this is separable from executive dysfunction and/or dementia. This study sought to determine whether distinct cognitive factors could be measured in a large PD cohort, and if those factors were differentially associated with other PD-related features, specifically to provide insight into visuospatial dysfunction. Methods: Non-demented participants with PD from the Pacific Udall Center were enrolled (n = 197). Co-participants (n = 104) completed questionnaires when available. Principal components factor analysis (PCFA) was utilized to group the neuropsychological test scores into independent factors by considering those with big factor loading (≥.40). Linear and logistic regression analyses were performed to examine the relationship between the cognitive factors identified in the PCFA and other clinical features of PD. Results: Six factors were extracted from the PCFA: 1) executive/processing speed, 2) visual learning & memory/visuospatial, 3) auditory working memory, 4) contextual verbal memory, 5) semantic learning & memory, and 6) visuospatial. Motor severity (p = 0.001), mood (p < 0.001), and performance on activities of daily living scores (informant: p < 0.001, patient: p = 0.009) were primarily associated with frontal and executive factors. General sleep disturbance (p < 0.006) and hallucinations (p = 0.002) were primarily associated with visuospatial functioning and visual learning/memory. Conclusions: Motor symptoms, mood, and performance on activities of daily living were primarily associated with frontal/executive factors. Sleep disturbance and hallucinations were associated with visuospatial functioning and visual learning/memory only, over and above executive functioning and regardless of cognitive disease severity. These findings support that visuospatial function in PD may indicate a more severe disease course, and that symptom management should be guided accordingly.


Drugs that reverse disease transcriptomic signatures are more effective in a mouse model of dyslipidemia.
High-throughput omics have proven invaluable in studying human disease, and yet day-to-day clinical practice still relies on physiological, non-omic markers. The metabolic syndrome, for example, is diagnosed and monitored by blood and urine indices such as blood cholesterol levels. Nevertheless, the association between the molecular and the physiological manifestations of the disease, especially in response to treatment, has not been investigated in a systematic manner. To this end, we studied a mouse model of diet-induced dyslipidemia and atherosclerosis that was subject to various drug treatments relevant to the disease in question. Both physiological data and gene expression data (from the liver and white adipose) were analyzed and compared. We find that treatments that restore gene expression patterns to their norm are associated with the successful restoration of physiological markers to their baselines. This holds in a tissue-specific manner—treatments that reverse the transcriptomic signatures of the disease in a particular tissue are associated with positive physiological effects in that tissue. Further, treatments that introduce large non-restorative gene expression alterations are associated with unfavorable physiological outcomes. These results provide a sound basis to in silico methods that rely on omic metrics for drug repurposing and drug discovery by searching for compounds that reverse a disease's omic signatures. Moreover, they highlight the need to develop drugs that restore the global cellular state to its healthy norm rather than rectify particular disease phenotypes.


Neuroadhesive L1 coating attenuates acute microglial attachment to neural electrodes as revealed by live two-photon microscopy.
Implantable neural electrode technologies for chronic neural recordings can restore functional control to paralysis and limb loss victims through brain-machine interfaces. These probes, however, have high failure rates partly due to the biological responses to the probe which generate an inflammatory scar and subsequent neuronal cell death. L1 is a neuronal specific cell adhesion molecule and has been shown to minimize glial scar formation and promote electrode-neuron integration when covalently attached to the surface of neural probes. In this work, the acute microglial response to L1-coated neural probes was evaluated in vivo by implanting coated devices into the cortex of mice with fluorescently labeled microglia, and tracking microglial dynamics with multi-photon microscopy for the ensuing 6 h in order to understand L1's cellular mechanisms of action. Microglia became activated immediately after implantation, extending processes towards both L1-coated and uncoated control probes at similar velocities. After the processes made contact with the probes, microglial processes expanded to cover 47.7% of the control probes' surfaces. For L1-coated probes, however, there was a statistically significant 83% reduction in microglial surface coverage. This effect was sustained through the experiment. At 6 h post-implant, the radius of microglia activation was reduced for the L1 probes by 20%, shifting from 130.0 to 103.5 μm with the coating. Microglia as far as 270 μm from the implant site displayed significantly lower morphological characteristics of activation for the L1 group. These results suggest that the L1 surface treatment works in an acute setting by microglial mediated mechanisms.


Cytokinin signalling inhibitory fields provide robustness to phyllotaxis.
How biological systems generate reproducible patterns with high precision is a central question in science. The shoot apical meristem (SAM), a specialized tissue producing plant aerial organs, is a developmental system of choice to address this question. Organs are periodically initiated at the SAM at specific spatial positions and this spatiotemporal pattern defines phyllotaxis. Accumulation of the plant hormone auxin triggers organ initiation, whereas auxin depletion around organs generates inhibitory fields that are thought to be sufficient to maintain these patterns and their dynamics. Here we show that another type of hormone-based inhibitory fields, generated directly downstream of auxin by intercellular movement of the cytokinin signalling inhibitor ARABIDOPSIS HISTIDINE PHOSPHOTRANSFER PROTEIN 6 (AHP6), is involved in regulating phyllotactic patterns. We demonstrate that AHP6-based fields establish patterns of cytokinin signalling in the meristem that contribute to the robustness of phyllotaxis by imposing a temporal sequence on organ initiation. Our findings indicate that not one but two distinct hormone-based fields may be required for achieving temporal precision during formation of reiterative structures at the SAM, thus indicating an original mechanism for providing robustness to a dynamic developmental system.


Influenza immunisation uptake in carers.
In 2005 the United Kingdom departments of health added 'carers' to the list of people that should be offered seasonal influenza immunisation by their general practice. We surveyed a sample of carers registered for care assistance with the charity Crossroads Caring for Carers. Over half (58%) were not aware that they are eligible for free influenza immunisation. Young carers without a chronic disease, were least likely to be offered immunisation and least likely to be immunised.


Differential localization of myosin-II isozymes in human cultured cells and blood cells.
We used purified polyclonal antibodies to human cytoplasmic myosin-IIA and myosin-IIB directly labeled with fluorescent dyes to localize these myosin-II isozymes in HeLa cells, melanoma cells and blood cells. Both antibodies react strongly with myosin-II isozymes in HeLa cells, melanoma cells and blood eosinophils, but only anti-myosin-IIA antibodies stain platelets, lymphocytes, neutrophils and monocytes in smears of human blood. Both antibodies stain small spots along the stress fibers of interphase HeLa cells and melanoma cells, but double staining revealed that the detailed distributions of myosin-IIA and myosin-IIB differ. A low concentration of diffuse myosin-IIB is present in the cortex, both in lamellar regions around the periphery of the cell and over the free surface. Myosin-IIB is also concentrated in spots along perinuclear stress fibers. Myosin-IIA is absent from the cortex but is concentrated in spots along stress fibers located near the basal surface of cultured cells. This population of peripheral stress fibers is highly enriched in myosin-IIA relative to myosin-IIB, but both are found together in centrally located stress fibers. In prophase and metaphase both isozymes are concentrated in the cortex in small spots less than 04.micron in size, similar to those in stress fibers. As the chromosomes begin the separate at anaphase, most of the myosin-II spots become concentrated in the outer 0.7 micron of the equatorial cortex in 100% of cells. This concentration of myosin-II isozymes in the cleavage furrow is maintained until the daughter cells separate. The superimposition of these small spots concentrated in the cleavage furrow produces the intense, uniform staining observed in conventional micrographs of whole cells.


Beyond 'vulnerable groups': contexts and dynamics of vulnerability.
This paper reviews approaches to vulnerability in public health, introducing a series of 10 papers addressing vulnerability in health in Africa. We understand vulnerability as simultaneously a condition and a process. Social inequalities are manifest in and exacerbate three key dimensions of vulnerability: the initial level of wellbeing, the degree of exposure to risk, and the capacity to manage risk effectively. We stress the dynamic interactions linking material and social deprivation, poverty, powerlessness and ill health: risks or shocks and their health impacts are intimately interconnected and reinforce each other in a cycle which in the absence of effective interventions, increases vulnerability. An inductive process which does not begin with an a priori definition or measurement of 'vulnerability' and which does not assume the existence of fixed 'vulnerable groups' allowed us both to re-affirm core aspects of existing conceptual frameworks, and to engage in new ways with literature specifically addressing vulnerability and resilience at the population level as well as with literature - for example in ecology, and on the concept of frailty in research on aging - with which researchers on health and poverty in Africa may not be familiar. We invite conceptual and empirical work on vulnerability in complex systems frameworks. These perspectives emphasize contexts and nonlinear causality thus supporting analyses of vulnerability and resilience as both markers and emergent properties of dynamic interactions. We accept a working definition of vulnerability, and recognize that some definable groups of people are more likely than others to suffer harm from exposure to health risks. But we suggest that the real work - at both intellectual and policy/political levels - lies in understanding and responding to the dynamics, meanings and power relations underlying actual instances and processes of vulnerability and harm.


To co-author or not to co-author: how to write, publish, and negotiate issues of authorship with undergraduate research students.
This Teaching Resource emphasizes the value of publishing with undergraduates and may be particularly helpful to incoming faculty who are new to the process of working with students. Beyond simply extolling the virtues of undergraduate research, we examine how such deep learning experiences for students can translate into unique opportunities for the faculty to demonstrate devotion to both teaching and scholarship. Along with highlighting the reasons faculty should consider publishing with undergraduates, we identify the particular challenges that accompany this suggestion and discuss strategies for overcoming them. Our resource includes two decision trees for helping faculty determine whether publishing with undergraduates represents a reasonable and attainable goal and whether an undergraduate has earned authorship. Based on our experience at primarily undergraduate institutions, we provide a list of strategies that may facilitate writing with undergraduates and lead to certain milestones in the careers of both students and faculty.


Duplicated genes with split functions: independent roles of protocadherin15 orthologues in zebrafish hearing and vision.
In the sensory receptors of both the eye and the ear, specialized apical structures have evolved to detect environmental stimuli such as light and sound. Despite the morphological divergence of these specialized structures and differing transduction mechanisms, the receptors appear to rely in part on a shared group of genes for function. For example, mutations in Usher (USH) genes cause a syndrome of visual and acoustic-vestibular deficits in humans. Several of the affected genes have been identified, including the USH1F gene, which encodes protocadherin 15 (PCDH15). Pcdh15 mutant mice also have both auditory and vestibular defects, although visual defects are not evident. Here we show that zebrafish have two closely related pcdh15 genes that are required for receptor-cell function and morphology in the eye or ear. Mutations in pcdh15a cause deafness and vestibular dysfunction, presumably because hair bundles of inner-ear receptors are splayed. Vision, however, is not affected in pcdh15a mutants. By contrast, reduction of pcdh15b activity using antisense morpholino oligonucleotides causes a visual defect. Optokinetic and electroretinogram responses are reduced in pcdh15b morpholino-injected larvae. In electron micrographs, morphant photoreceptor outer segments are improperly arranged, positioned perpendicular to the retinal pigment epithelium and are clumped together. Our results suggest that both cadherins act within their respective transduction organelles: Pcdh15a is necessary for integrity of the stereociliary bundle, whereas Pcdh15b is required for alignment and interdigitation of photoreceptor outer segments with the pigment epithelium. We conclude that after a duplication of pcdh15, one gene retained an essential function in the ear and the other in the eye.


Modulating carnitine levels by targeting its biosynthesis pathway - selective inhibition of γ-butyrobetaine hydroxylase.
Carnitine is essential for fatty acid metabolism, but is associated with both health benefits and risks, especially heart diseases. We report the identification of potent, selective and cell active inhibitors of γ-butyrobetaine hydroxylase (BBOX), which catalyses the final step of carnitine biosynthesis in animals. A crystal structure of BBOX in complex with a lead inhibitor reveals that it binds in two modes, one of which adopts an unusual 'U-shape' conformation stabilised by inter- and intra-molecular π-stacking interactions. Conformational changes observed on binding of the inhibitor to BBOX likely reflect those occurring in catalysis; they also rationalise the inhibition of BBOX by high levels of its substrate γ-butyrobetaine (GBB), as observed both with isolated BBOX protein and in cellular studies.


Developmental regulation of human cortex transcription and its clinical relevance at single base resolution.
Transcriptome analysis of human brain provides fundamental insight into development and disease, but it largely relies on existing annotation. We sequenced transcriptomes of 72 prefrontal cortex samples across six life stages and identified 50,650 differentially expression regions (DERs) associated with developmental and aging, agnostic of annotation. While many DERs annotated to non-exonic sequence (41.1%), most were similarly regulated in cytosolic mRNA extracted from independent samples. The DERs were developmentally conserved across 16 brain regions and in the developing mouse cortex, and were expressed in diverse cell and tissue types. The DERs were further enriched for active chromatin marks and clinical risk for neurodevelopmental disorders such as schizophrenia. Lastly, we demonstrate quantitatively that these DERs associate with a changing neuronal phenotype related to differentiation and maturation. These data show conserved molecular signatures of transcriptional dynamics across brain development, have potential clinical relevance and highlight the incomplete annotation of the human brain transcriptome.


International prospective observational study on intracranial pressure in intensive care (ICU): the SYNAPSE-ICU study protocol.
Intracranial pressure (ICP) monitoring is commonly used in neurocritical care patients with acute brain injury (ABI). Practice about indications and use of ICP monitoring in patients with ABI remains, however, highly variable in high-income countries, while data on ICP monitoring in low and middle-income countries are scarce or inconsistent. The aim of the SYNAPSE-ICU study is to describe current practices of ICP monitoring using a worldwide sample and to quantify practice variations in ICP monitoring and management in neurocritical care ABI patients. The SYNAPSE-ICU study is a large international, prospective, observational cohort study. From March 2018 to March 2019, all patients fulfilling the following inclusion criteria will be recruited: age >18 years; diagnosis of ABI due to primary haemorrhagic stroke (subarachnoid haemorrhage or intracranial haemorrhage) or traumatic brain injury; Glasgow Coma Score (GCS) with no eye opening (Eyes response=1) and Motor score ≤5 (not following commands) at ICU admission, or neuro-worsening within the first 48 hours with no eye opening and a Motor score decreased to ≤5. Data related to clinical examination (GCS, pupil size and reactivity, Richmond Agitation-Sedation Scale score, neuroimaging) and to ICP interventions (Therapy Intensity Levels) will be recorded on admission, and at day 1, 3 and 7. The Glasgow Outcome Scale Extended (GOSE) will be collected at discharge from ICU and from hospital and at 6-month follow-up. The impact of ICP monitoring and ICP-driven therapy on GOSE will be analysed at both patient and ICU level. The study has been approved by the Ethics Committee 'Brianza' at the Azienda Socio Sanitaria Territoriale (ASST)-Monza (approval date: 21 November 2017). Each National Coordinator will notify the relevant ethics committee, in compliance with the local legislation and rules. Data will be made available to the scientific community by means of abstracts submitted to the European Society of Intensive Care Medicine annual conference and by scientific reports and original articles submitted to peer-reviewed journals. NCT03257904.


Suppression of Zeeman splitting of the energy levels of exciton-polariton condensates in semiconductor microcavities in an external magnetic field.
A key property of equilibrium exciton-polariton condensates in semiconductor microcavities is the suppression of the Zeeman splitting under a magnetic field. By studying magnetophotoluminescence spectra from a GaAs microcavity, we show experimentally that a similar effect occurs in a nonequilibrium polariton condensate arising from polariton parametric scattering. In this case, the quenching of Zeeman splitting is related to a phase synchronization of spin-up and spin-down polarized polariton condensates caused by a nonlinear coupling via the coherent pump state.


Receptor activator of NF-kappaB (RANK) ligand induces ectodomain shedding of RANK in murine RAW264.7 macrophages.
Osteoclastogenesis is a highly sophisticated process that involves a variety of membrane-bound proteins expressed in osteoblasts and osteoclast precursors. Over the past several years, proteolytic cleavage and release of the ectodomain of membrane-bound proteins, also referred to as ectodomain shedding, has emerged as an important posttranslational regulatory mechanism for modifying the function of cell surface proteins. In line with this notion, several membrane-bound molecules involved in osteoclastogenesis, including CSF-1R and receptor activator of NF-kappaB ligand (RANKL), are proteolytically cleaved and released from the cell surface. In this study, we investigated whether receptor activator of NF-kappaB (RANK), one of the most essential molecules in osteoclastogenesis, undergoes ectodomain shedding. The results showed that RANK is released in the form of a soluble monomeric protein and that TNF-alpha-converting enzyme is involved in this activity. We also identified potential cleavage sites in the juxtamembrane domain of RANK and found that rRANKL induces RANK shedding in a macrophage-like cell line RAW264.7 via TNFR-associated factor 6 and MAPK pathways. Furthermore, we found that RANKL-induced osteoclastogenesis is accelerated in TNF-alpha-converting enzyme-deficient osteoclast precursors. These observations suggest the potential involvement of ectodomain shedding in the regulation of RANK functions and may provide novel insights into the mechanisms of osteoclastogenesis.


Evidence for mid-Holocene rice domestication in the Americas.
The development of agriculture is one of humankind's most pivotal achievements. Questions about plant domestication and the origins of agriculture have engaged scholars for well over a century, with implications for understanding its legacy on global subsistence strategies, plant distribution, population health and the global methane budget. Rice is one of the most important crops to be domesticated globally, with both Asia (Oryza sativa L.) and Africa (Oryza glaberrima Steud.) discussed as primary centres of domestication. However, until now the pre-Columbian domestication of rice in the Americas has not been documented. Here we document the domestication of Oryza sp. wild rice by the mid-Holocene residents of the Monte Castelo shell mound starting at approximately 4,000 cal. yr BP, evidenced by increasingly larger rice husk phytoliths. Our data provide evidence for the domestication of wild rice in a region of the Amazon that was also probably the cradle of domestication of other major crops such as cassava (Manihot esculenta), peanut (Arachis hypogaea) and chilli pepper (Capsicum sp.). These results underline the role of wetlands as prime habitats for plant domestication worldwide.


The role of space charge formation in the generation of thermally stimulated current (TSC) spectroscopy data for a model amorphous drug system.
Thermally stimulated current (TSC) spectroscopy is attracting increasing attention as a means of materials characterization, particularly in terms of measuring slow relaxation processes in solid samples. However, wider use of the technique within the pharmaceutical field has been inhibited by difficulties associated with the interpretation of TSC data, particularly in terms of deconvoluting dipolar relaxation processes from charge distribution phenomena. Here, we present evidence that space charge and electrode contact effects may play a significant role in the generation of peaks that have thus far proved difficult to interpret. We also introduce the use of a stabilization temperature in order to control the space charge magnitude. We have studied amorphous indometacin as a model drug compound and have varied the measurement parameters (stabilization and polarization temperatures), interpreting the changes in spectral composition in terms of charge redistribution processes. More specifically, we suggested that charge drift and diffusion processes, charge injection from the electrodes and high activation energy charge redistribution processes may all contribute to the appearance of shoulders and 'spurious' peaks. We present recommendations for eliminating or reducing these effects that may allow more confident interpretation of TSC data.


A nitrogen starvation-induced dormant G0 state in fission yeast: the establishment from uncommitted G1 state and its delay for return to proliferation.
Fission yeast cells either remain in the mitotic cell cycle or exit to meiotic sporulation from an uncommitted G1 state dependent on the presence or absence of nitrogen source in the medium (Nurse and Bissett, 1981). We examined how heterothallic haploid cells, which cannot sporulate, behave under nitrogen-starvation for longer than 25 days at 26 degrees C. These cells were shown to enter a stable state (designated the dormant G0) with nearly full viability. Maintaining the dormant cells required glucose, suggesting that the cells remained metabolically active although cell division had ceased. They differed dramatically from mitotic and uncommitted G1 cells in heat resistance, and also in cytoplasmic and nuclear morphologies. After nitrogen replenishment, the initial responses of dormant G0 cells were investigated. The kinetics for reentry into the proliferative state were delayed considerably, and the changes in cell shape were enhanced particularly for those recovering from extended nitrogen starvation. A part of the delay could be accounted for by the duration of nuclear decondensation and cell elongation for the first cell division.