Notch Signaling Activates Stem Cell Properties of Müller Glia through Transcriptional Regulation and Skp2-mediated Degradation of p27Kip1.
Müller glia (MG), the sole glial cells generated by retinal progenitors, have emerged as a viable cellular target for therapeutic regeneration in degenerative blinding diseases, as they possess dormant stem cell properties. However, the mammalian MG does not display the neurogenic potential of their lower vertebrate counterparts, precluding their practical clinical use. The answer to this barrier may be found in two interlinked processes underlying the neurogenic potential, i.e., the activation of the dormant stem cell properties of MG and their differentiation along the neuronal lineage. Here, we have focused on the former and examined Notch signaling-mediated activation of MG. We demonstrate that one of the targets of Notch signaling is the cyclin-dependent kinase inhibitor (CKI), p27Kip1, which is highly expressed in quiescent MG. Notch signaling facilitates the activation of MG by inhibiting p27Kip1 expression. This is likely achieved through the Notch- p27Kip1 and Notch-Skp2-p27Kip1 axes, the former inhibiting the expression of p27Kip1 transcripts and the latter levels of p27Kip1 proteins by Skp2-mediated proteasomal degradation. Thus, Notch signaling may facilitate re-entry of MG into the cell cycle by inhibiting p27Kip1 expression both transcriptionally and post-translationally.


Bulk phase behavior and interfacial properties of binary mixtures of Lennard-Jones chains and solvent: a density functional theory study.
A Density Functional Theory study is performed to analyze both bulk and interfacial properties of solvent-polymer binary mixtures. The effects of increasing polymer chain length on the bulk phase diagram morphology and interfacial tension are presented and compared to the prior simulation results. Good agreement between simulation and Density Functional Theory is found, including its ability to reproduce the density inversion phenomenon for highly asymmetric solvent-polymer binary mixtures. The data on the interfacial tensions is used to compute contact angles of the mixture at a planar wall, with particular focus on the wetting transition. The dependence of the wetting temperature on the polymer chain length and the mixture composition is analyzed in detail.


A comparison of episodic heart-rate patterns in the fetus and newborn.
Fourier analysis was applied to episodes of high and low heart-rate variation in the near-term fetus. The spectral densities of the variations in pulse intervals from 1 to 7 cycles/min, were almost identical with those found by De Haan et al. (1977) in the newborn infant during both active and quiet sleep. A computer programme, designed to separate low and high heart-rate variation episodes in the fetus, identified seven of the eight quiet sleep episodes in four infants. None of the other neonatal behavioural states had low heart-rate variation. It was concluded that, with other published data, there is increasing, although indirect, evidence that low heart-rate variation episodes indicate quiet sleep in the human fetus near term.


Role of echocardiography in differential diagnosis of broad complex tachycardia.
It is not always easy to distinguish between supraventricular tachycardia with aberration and ventricular tachycardia by electrocardiographic analysis alone. M mode echocardiography can often help by providing direct or indirect evidence of the relation between atrial and ventricular contraction. Sixteen consecutive patients with spontaneous sustained broad QRS complex tachycardia with heart rates of 120-225 beats/minute were examined. Echocardiographic evidence of 1:1 conduction was seen in three cases and 2:1 atrioventricular conduction in one (all four had supraventricular tachycardia, confirmed by intracardiac electrocardiography in three). Evidence of retrograde block was seen in 12 (all had ventricular tachycardia, with electrophysiological confirmation in 10). Satisfactory views of the mitral valve were obtained in all patients. Patients with ventricular tachycardia had a variable mitral valve opening time (range 42-110%) compared with those who had supraventricular tachycardia (9-15%). Aortic root and left atrial views gave direct evidence of atrial contraction in three cases, and subcostal right atrial wall views were diagnostic in four of five cases. Seven patients with ventricular tachycardia had been wrongly diagnosed elsewhere as having supraventricular tachycardia. This study confirms that echocardiography is a simple and rapid aid to accurate diagnosis in patients with broad QRS complex tachycardia.


Electroluminescent Pressure-Sensing Displays.
Simultaneous sensing and visualization of pressure provides a useful platform to obtain information about a pressurizing object, but the fabrication of such interactive displays at the single-device level remains challenging. Here, we present a pressure responsive electroluminescent (EL) display that allows for both sensing and visualization of pressure. Our device is based on a two-terminal capacitor with six constituent layers: top electrode/insulator/hole injection layer/emissive layer/electron transport layer/bottom electrode. Light emission upon exposure to an alternating current field between two electrodes is controlled by the capacitance change of the insulator arising from the pressure applied on top. Besides capacitive pressure sensing, our EL display allows for direct visualization of the static and dynamic information of position, shape, and size of a pressurizing object on a single-device platform. Monitoring the pressurized area of an elastomeric hemisphere on a device by EL enables quantitative estimation of the Young's modulus of the elastomer, offering a new and facile characterization method for the mechanical properties of soft materials.


Proteomics investigation on aristolochic acid nephropathy: a case study on rat kidney tissues.
Prolonged intake of aristolochic acid (AA) has been shown to be associated with the development of certain renal disorders. Renal tubular atrophy and interstitial fibrosis are the early symptoms of AA nephropathy. The symptoms were observed in rats that were dosed with AA at a dosage of 10 mg/kg/day for 1 month. Apart from the renal tubular atrophy and interstitial fibrosis, AA-DNA adducts were detected in the rat kidney tissue. Differentiated proteins were identified in the kidney tissues from proteomics investigations. The upregulated proteins identified included ornithine aminotransferase, sorbitol dehydrogenase, actin, aspartoacylase, 3-hydroxyisobutyrate dehydrogenase, and peroxiredoxin-1. Downregulated proteins such as ATP synthase subunit β, glutamate dehydrogenase 1, regucalcin, glutamate-cysteine ligase regulatory subunit, dihydropteridine reductase, hydroxyacyl-coenzyme A dehydrogenase, voltage-dependent anion-selective channel protein 1, prohibitin, and adenylate kinase isoenzyme 4 were also identified. Several identified protein markers were found to have biological and medical significance.


Pillar[5]arene-Based Fluorescent Supramolecular Polymers Without Conventional Chromophores.
Fluorescent supramolecular polymers have garnered significant attention due to their successful integration of supramolecular polymers and fluorescence, offering vast potential for applications in sensing, imaging, optoelectronics, and photonics. In this study, we present a novel supramolecular polymer based on P5-OH, derived from mono-substituted pillararene macrocycles. Notably, these formed supramolecular polymeric aggregates exhibit a prominent blue emission, representing a rare instance of fluorescent polymers devoid of conventional chromophores. Furthermore, through the modification of alkyl chain ending groups attached to pillar[5]arenes, slight shifts in the emission peak could be observed. This research expands the scope of functional supramolecular polymeric systems utilizing pillararenes, providing valuable insights for the design of innovative luminescent materials and optical devices.


Cosurfactant effect of a semifluorinated alkane at a fluorocarbon/water interface: impact on the stabilization of fluorocarbon-in-water emulsions.
Previous work has demonstrated that semifluorinated alkanes CnF2n+1CmH2m+1 (FnHm diblocks), when used in conjunction with phospholipids, strongly stabilize fluorocarbon (FC)-in-water emulsions destined to be used as oxygen carriers. Although the presence of FnHm diblocks in the emulsion's interfacial phospholipid film was suggested to account for the observed stabilization, no direct proof of the diblock's location has been provided so far. We now report definite experimental evidence of the diblock's presence at the interfacial film, both on a macroscopic level by investigating the FC/water interface using the pendant drop method and directly on emulsions by monitoring their stability for various phospholipid chain lengths. We first establish that F8H16 has a strong cosurfactant effect with phospholipids [dimyristoylphosphatidylcholine (DMPC), dilaurylphosphatidylcholine (DLPC), dioctanoylphosphatidylcholine (PCL8)] at a perfluorooctyl bromide (PFOB)/water interface, as evidenced by a dramatic F8H16-concentration-dependent decrease of the interfacial tension. Where FC emulsions are concerned, we show that the stabilization effect, which consists of a decrease of the rate of molecular diffusion of the FC, depends strongly on the length of the phospholipid's fatty chain as compared to the length of the hydrocarbon segment, Hm, of the diblock. Stabilization is maximized when the Hm length is similar to that of the phospholipid's fatty chains. A strong mismatch between Hm and the phospholipid chain length can actually destabilize the emulsion. A different destabilization mechanism is then at work: coalescence. The presence of F8H16 at the interfacial film is further supported by the fact that perfluorodecyl bromide, a heavy analogue of PFOB that stabilizes PFOB emulsions by lowering the solubility and diffusibility of the emulsion's dispersed FC phase, exercises its stabilizing effect similarly for all the phospholipids investigated.


Evaluating patient acceptability of a culturally focused psychiatric consultation intervention for Latino Americans with depression.
Significant disparities exist in both access to and quality of mental health care for Latino Americans with depression compared to Caucasians, resulting in a greater burden of disability in this underserved population. Our aim is to evaluate participant acceptability of a Culturally Focused Psychiatric (CFP) consultation program for depressed Latino Americans. Latino American adult primary care patients endorsing depressive symptoms on a screening questionnaire were targeted in their primary care clinic. The intervention addressed participants' depressive symptoms using culturally adapted clinical assessments and toolkits. Acceptability was evaluated using a treatment satisfaction scale and in-depth semi-structured interviews. Overall, 85% of participants responded positively to all questions of the satisfaction scale. In in-depth interviews, the vast majority of participants reported the program met expectations, all stated providers were culturally sensitive, and most stated recommendations were culturally sensitive. The CFP program was found to be acceptable to a group of depressed Latino American primary care patients. Further research is needed to evaluate if the CFP intervention can improve depressive symptoms and outcomes.


Identification of a cytoplasm to vacuole targeting determinant in aminopeptidase I.
Aminopeptidase I (API) is a soluble leucine aminopeptidase resident in the yeast vacuole (Frey, J., and K.H. Rohm. 1978. Biochim. Biophys. Acta. 527:31-41). The precursor form of API contains an amino-terminal 45-amino acid propeptide, which is removed by proteinase B (PrB) upon entry into the vacuole. The propeptide of API lacks a consensus signal sequence and it has been demonstrated that vacuolar localization of API is independent of the secretory pathway (Klionsky, D.J., R. Cueva, and D.S. Yaver. 1992. J. Cell Biol. 119:287-299). The predicted secondary structure for the API propeptide is composed of an amphipathic alpha-helix followed by a beta-turn and another alpha-helix, forming a helix-turn-helix structure. With the use of mutational analysis, we determined that the API propeptide is essential for proper transport into the vacuole. Deletion of the entire propeptide from the API molecule resulted in accumulation of a mature-sized protein in the cytosol. A more detailed examination using random mutagenesis and a series of smaller deletions throughout the propeptide revealed that API localization is severely affected by alterations within the predicted first alpha-helix. In vitro studies indicate that mutations in this predicted helix prevent productive binding interactions from taking place. In contrast, vacuolar import is relatively insensitive to alterations in the second predicted helix of the propeptide. Examination of API folding revealed that mutations that affect entry into the vacuole did not affect the structure of API. These data indicate that the API propeptide serves as a vacuolar targeting determinant at a critical step along the cytoplasm to vacuole targeting pathway.


Analysis of agonist function at fusion proteins between the IP prostanoid receptor and cognate, unnatural and chimaeric G-proteins.
Direct measures of G-protein activation based on guanine nucleotide exchange and hydrolysis are frequently impossible to monitor for receptors which interact predominantly with G(s)alpha. An isolated FLAG (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys)-epitope-tagged human IP prostanoid receptor and fusion proteins generated between this form of the receptor and the alpha subunits of its cognate G-protein G(s), G(i1), a G-protein which it fails to activate in co-expression studies, and a chimaeric G(i1)-G(s)6 (a form of G(i1) in which the C-terminal six amino acids were replaced with the equivalent sequence of G(s)) were stably expressed in HEK293 cells. These were detected by [(3)H]ligand-binding studies and by immunoblotting with both an anti-FLAG antibody and with appropriate antisera to the G-proteins. Each construct displayed similar affinity to bind the agonist iloprost. Iloprost stimulated adenylate cyclase activity in clones expressing both IP prostanoid receptor and the IP prostanoid receptor-G(s)alpha fusion protein, and both constructs were shown to interact with and activate endogenously expressed G(s)alpha. Addition of iloprost to membranes of cells expressing the isolated receptor resulted in a small stimulation of high-affinity GTPase activity. Iloprost produced no stimulation of GTPase activity which could be attributed to the IP prostanoid receptor-G(i1)alpha fusion. However, the fusion proteins containing either G(s)alpha or G(i1)-G(s)6alpha produced substantially greater stimulation of GTPase activity than the isolated IP prostanoid receptor. Treatment of cells expressing the IP prostanoid receptor-G(i1)-G(s)6alpha fusion protein with a combination of cholera and pertussis toxins allowed direct measurement of agonist activation of the receptor-linked G-protein. Normalization of such results for levels of expression of the IP prostanoid receptor constructs demonstrated a 5-fold higher stimulation of GTPase activity when using the G(s)alpha-containing fusion protein and a 9-fold improvement when using the fusion protein containing G(i1)-G(s)6alpha to detect G-protein activation compared with expression of the isolated receptor.


[Several subclinical forms of epileptic dysphoria and their ambulatory treatment].
As a result of a clinico-biochemical study covering 58 epileptic patients with dysphoric states, the authors identified 2 clinical groups with the characteristic clinical features and biochemical profile. On the basis of the data obtained, a definite tactics of the treatment for dysphoric states is proposed. Multiple modality treatment with the use of anticonvulsants, neuroleptics, antidepressants and psychostimulants is recommended.


Effects of nurse-to-patient ratio legislation on nurse staffing and patient mortality, readmissions, and length of stay: a prospective study in a panel of hospitals.
Substantial evidence indicates that patient outcomes are more favourable in hospitals with better nurse staffing. One policy designed to achieve better staffing is minimum nurse-to-patient ratio mandates, but such policies have rarely been implemented or evaluated. In 2016, Queensland (Australia) implemented minimum nurse-to-patient ratios in selected hospitals. We aimed to assess the effects of this policy on staffing levels and patient outcomes and whether both were associated. For this prospective panel study, we compared Queensland hospitals subject to the ratio policy (27 intervention hospitals) and those that discharged similar patients but were not subject to ratios (28 comparison hospitals) at two timepoints: before implementation of ratios (baseline) and 2 years after implementation (post-implementation). We used standardised Queensland Hospital Admitted Patient Data, linked with death records, to obtain data on patient characteristics and outcomes (30-day mortality, 7-day readmissions, and length of stay [LOS]) for medical-surgical patients and survey data from 17 010 medical-surgical nurses in the study hospitals before and after policy implementation. Survey data from nurses were used to measure nurse staffing and, after linking with standardised patient data, to estimate the differential change in outcomes between patients in intervention and comparison hospitals, and determine whether nurse staffing changes were related to it. We included 231 902 patients (142 986 in intervention hospitals and 88 916 in comparison hospitals) assessed at baseline (2016) and 257 253 patients (160 167 in intervention hospitals and 97 086 in comparison hospitals) assessed in the post-implementation period (2018). After implementation, mortality rates were not significantly higher than at baseline in comparison hospitals (adjusted odds ratio [OR] 1·07, 95% CI 0·97-1·17, p=0·18), but were significantly lower than at baseline in intervention hospitals (0·89, 0·84-0·95, p=0·0003). From baseline to post-implementation, readmissions increased in comparison hospitals (1·06, 1·01-1·12, p=0·015), but not in intervention hospitals (1·00, 0·95-1·04, p=0·92). Although LOS decreased in both groups post-implementation, the reduction was more pronounced in intervention hospitals than in comparison hospitals (adjusted incident rate ratio [IRR] 0·95, 95% CI 0·92-0·99, p=0·010). Staffing changed in hospitals from baseline to post-implementation: of the 36 hospitals with reliable staffing measures, 30 (83%) had more than 4·5 patients per nurse at baseline, with the number decreasing to 21 (58%) post-implementation. The majority of change was at intervention hospitals, and staffing improvements by one patient per nurse produced reductions in mortality (OR 0·93, 95% CI 0·86-0·99, p=0·045), readmissions (0·93, 0·89-0·97, p<0·0001), and LOS (IRR 0·97, 0·94-0·99, p=0·035). In addition to producing better outcomes, the costs avoided due to fewer readmissions and shorter LOS were more than twice the cost of the additional nurse staffing. Minimum nurse-to-patient ratio policies are a feasible approach to improve nurse staffing and patient outcomes with good return on investment. Queensland Health, National Institutes of Health, National Institute of Nursing Research.


Histone macroH2A1 is concentrated in the inactive X chromosome of female preimplantation mouse embryos.
MacroH2As are core histone proteins with a hybrid structure consisting of a domain that closely resembles a full-length histone H2A followed by a large nonhistone domain. We recently showed that one of the macroH2A subtypes, macroH2A1.2, is concentrated in the inactive X chromosome in adult female mammals. Here we examine the timing of the association of macroH2A1.2 with the inactive X chromosome during preimplantation mouse development in order to assess the possibility that macroH2A1 participates in the initiation of X inactivation. The association of macroH2A1.2 with one of the X chromosomes was observed in 50% of blastocysts, occurring mostly, if not exclusively, in extraembryonic cells as was expected from previous studies, which indicated that X inactivation in embryonic lineages happens after implantation. Examination of earlier embryonic stages indicates that the association of macroH2A1 with the inactive X chromosome begins between the 8- and 16-cell stages. Of the changes that are known to happen during X inactivation in preimplantation embryos, the accumulation of macroH2A1 appears to be the earliest marker of the inactive X chromosome and is the only change that has been shown to occur during the period when transcriptional silencing is initiated.


Classification of Hemilabile Ligands Using Machine Learning.
Hemilabile ligands have the capacity to partially disengage from a metal center, providing a strategy to balance stability and reactivity in catalysis, but they are not straightforward to identify. We identify ligands in the Cambridge Structural Database that have been crystallized with distinct denticities and are thus identifiable as hemilabile ligands. We implement a semi-supervised learning approach using a label-spreading algorithm to augment a small negative set that is supported by heuristic rules of ligand and metal co-occurrence. We show that a heuristic based on coordinating atom identity alone is not sufficient to identify whether a ligand is hemilabile, and our trained machine-learning classification models are instead needed to predict whether a bi-, tri-, or tetradentate ligand is hemilabile with high accuracy and precision. Feature importance analysis of our models shows that the second, third, and fourth coordination spheres all play important roles in ligand hemilability.


Self-reported experiences of everyday discrimination are associated with elevated C-reactive protein levels in older African-American adults.
Self-reported experiences of "everyday" discrimination have been linked to indices of cardiovascular disease and overall mortality and findings have been particularly pronounced for African-American populations. However, the biological mechanisms underlying these associations remain unclear. C-reactive protein (CRP), a marker of inflammation, is a known correlate of cardiovascular and other health outcomes and has also been linked to several psychosocial processes. To our knowledge, no studies have examined the association between experiences of discrimination and CRP. We examined the cross-sectional association between self-reported experiences of discrimination and CRP in a sample of 296 older African-American adults (70% female, mean age=73.1). Experiences of discrimination were assessed with the 9-item everyday discrimination scale and CRP was assayed from blood samples. In linear regression models adjusted for age, sex and education, experiences of discrimination were associated with higher levels of CRP (B=.10, p=.03). This association remained significant after additional adjustments for depressive symptoms (B=.10, p=.04), smoking, and chronic health conditions (heart disease, diabetes, hypertension) that might influence inflammation (B=.11, p=.02). However, results were attenuated when body mass index (BMI) was added to the model (B=.09, p=.07). In conclusion, self-reported experiences of everyday discrimination are associated with higher levels of CRP in older African-American adults, although this association is not completely independent of BMI.


Subcellular localization of the enzymes that dephosphorylate myo-inositol polyphosphates in human platelets.
The phosphatase-induced hydrolysis of [3H]inositol 1,4-bisphosphate [Ins(1,4)P2)] and [3H]inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] was studied in platelet subcellular fractions. The activity that hydrolyses Ins(1,4)P2 is cytosolic, whereas the activity that hydrolyses Ins(1,4,5)P3 is present in both particulate and cytosolic fractions. The cytosolic Ins(1,4)P2 phosphatase hydrolyses the 1-phosphate of Ins(1,4)P2, whereas the cytosolic and membrane-bound Ins(1,4,5)P3 phosphatases hydrolyse the 5-phosphate of Ins(1,4,5)P3. In the presence of ATP, it is possible to observe a cytosolic Ins(1,4,5)P3 3-kinase that phosphorylates Ins(1,4,5)P3 to inositol 1,3,4,5-tetrakisphosphate. Apparent Km values for the particulate and the cytosolic Ins(1,4,5)P3 phosphatases are 100 microM and 40 microM respectively. A large proportion of the membrane-associated Ins(1,4,5)P3 phosphatase can be extracted with 1 M-NaCl, and the Mr of this enzyme, as determined by hydrodynamic studies, is 49,000, whereas that of the cytosolic enzyme is 59,000. The Km values for the cytosolic Ins(1,4)P2 phosphatase is 40 microM; this enzyme has an Mr of 49,000. The highest specific activity of the Ins(1,4,5)P3 phosphatase is present in a highly purified plasma-membrane fraction.


Demonstrating polymorphic miRNA-mediated gene regulation in vivo: application to the g+6223G->A mutation of Texel sheep.
We herein describe the development of a biochemical method to evaluate the effect of single nucleotide polymorphisms (SNPs) in target genes on their regulation by microRNAs in vivo. The method is based on the detection of allelic imbalance in RNAs coimmunoprecipitated with AGO proteins from tissues of heterozygous individuals. We characterize the performances of our approach using a model system in a cell culture, and then apply it successfully to prove that the 3'UTR g+6223G-->A mutation operates by promoting RISC-dependent down-regulation of myostatin (MSTN) in skeletal muscle of Texel sheep.


Factors Associated With COVID-19 Vaccine Hesitancy in Rheumatology Outpatients in New York City.
The aim of this study was to measure COVID-19 vaccine hesitancy among rheumatology outpatients from an early COVID-19 "hotspot" during the initial period of vaccine availability. In March 2021, a Web-based survey was sent to 7505 adults seen at a Rheumatology Division in New York City. We evaluated characteristics associated with 3 categories of COVID-19 vaccination status: declined, undecided, and willing/already received. We used multinomial logistic regression models to calculate relative risk ratios assessing predictors of vaccination status. Among 2384 (32%) respondents (80% female, 87% White, 59% with systemic rheumatic disease), 2240 (94.0%) were willing/already received COVID-19 vaccination, 88 (3.7%) were undecided, and 56 (2.3%) declined. Compared with those willing/already vaccinated, those declining or undecided were younger, more likely identified as Black or Hispanic/Latinx, and had lower household income and educational attainment. Immunosuppressive medication use did not differ among groups. After multivariable adjustment, every 1-year increase in age was associated with a 0.96 lower relative risk of declining or being undecided versus willing/already vaccinated. Respondents identifying as Black versus White had a higher relative risk ratio of being undecided (4.29 [95% confidence interval, 1.96-9.36]), as did those identifying as Hispanic/Latinx versus non-Hispanic/non-Latinx (2.81 [95% confidence interval, 1.29-6.09]). Those declining vaccination were least likely to believe in general vaccine importance or the safety and efficacy of the COVID-19 vaccine. Among rheumatology patients in New York City with and without systemic rheumatic disease, COVID-19 vaccine uptake was high after its initial availability. Sociodemographic but not medication-related factors were associated with vaccine hesitancy; these findings can inform future rheumatology vaccination programs.


Intramacrophagic Mycobacterium avium bacilli are coated by a multiple lamellar structure: freeze fracture analysis of infected mouse liver.
We used freeze fracture electron microscopy to study the fine structure of Mycobacterium avium inside phagosomes of murine macrophages. M. avium-susceptible C57BL/6 mice were infected with M. avium by intraperitoneal inoculation of 10(8) viable bacilli. We studied the microanatomy of the mycobacteria in 3-month infections of mice, a situation in which bacillary multiplication is extensive. In these samples, freeze fracture revealed that intraphagosomal bacilli were surrounded by a multilamellar coat that was apposed to the cell wall. In thin sections, in contrast, the area corresponding to the coat showed no substructure and was electron transparent (the so-called electron-transparent zone that has been previously reported by others). The multiple lamellae resembled an onionlike assembly that was inserted in between the mycobacterial wall outer surface and the phagosomal membrane. Each lamella of the M. avium coat was made up of parallel straight fibrils with a width of 5 nm. A variable number of lamellae, sometimes up to 10 or more elements, coated individual bacilli. The multilamellar coat was absent around both extracellular M. avium and intramacrophagic M. avium after short-term (45-min) inoculation of mice. The supramolecular organization of the M. avium lamellar coat as viewed here by freeze fracture is similar to that of purified mycoside C (P. Draper, J. Gen. Microbiol. 83:431-433, 1974; K.-S. Kim, M.R.J. Salton, and L. Barksdale, J. Bacteriol. 125:739-743, 1976), a mycobacterial component currently known as glycopeptidolipid (W.W. Barrow and P.J. Brennan, J. Bacteriol. 150:381-384, 1982). We conclude that M. avium bacilli growing in macrophages are surrounded by multilamellar capsulelike structures that contain glycopeptidolipid molecules.


Serum Nestorone and ethinyl estradiol levels, and ovulation inhibition in women using three different dosage combinations of a Nestorone progestogen-ethinyl estradiol contraceptive vaginal ring on a bleeding-signaled regimen.
This trial tested the hypothesis that menstrually signaled use of contraceptive vaginal rings ("rings") releasing low-dose combinations of Nestorone (NES) and ethinyl estradiol (EE) would reliably suppress luteal activity and ovulation, and prevent unintended pregnancy, while controlling the incidence of menstrual bleeding episodes and bleeding days. Nestorone/ethinyl estradiol rings releasing 50/10, 50/20 and 150/15 mug/day were studied through 6 months. A ring was to be used continuously, until its removal was signaled by menstrual bleeding. Reinsertion was required 96 h after removal. Serum for NES, EE and progesterone were collected and assayed, and vaginal ultrasound scans were performed in three 5-week periods to examine luteal activity, follicular growth, ovulation and their correlates. In 10 subjects using the 150/15 ring, six samples were drawn in the 24-h period after ring removal to examine serum levels of NES and EE. One hundred sixty subjects at three doses provided blood samples. Median serum concentrations of NES and EE demonstrated dose ratios slightly below the nominal dose ratios expected. Serum NES concentrations declined 19-22% from weeks 3 to 25. Changes in EE levels depended on dose. Nestorone levels fell 81% by 24 h after ring removal and EE levels fell by 50%. Luteal activity was completely suppressed in 94-95% of cycles and in 90% of subjects. Three pregnancies occurred in subjects participating in this serum sampling study. Satisfactory serum levels of NES and EE, and a high level of ovulation suppression were achieved. Irregular ring use, however, permitted pregnancies to occur.


Human-mediated dispersal of seeds over long distances.
Human activities have fundamental impacts on the distribution of species through altered land use, but also directly by dispersal of propagules. Rare long-distance dispersal events have a disproportionate importance for the spread of species including invasions. While it is widely accepted that humans may act as vectors of long-distance dispersal, there are few studies that quantify this process. We studied in detail a mechanism of human-mediated dispersal (HMD). For two plant species we measured, over a wide range of distances, how many seeds are carried by humans on shoes. While over half of the seeds fell off within 5m, seeds were regularly still attached to shoes after 5 km. Semi-mechanistic models were fitted, and these suggested that long-distance dispersal on shoes is facilitated by decreasing seed detachment probability with distance. Mechanistic modelling showed that the primary vector, wind, was less important as an agent of long-distance dispersal, dispersing seeds less than 250 m. Full dispersal kernels were derived by combining the models for primary dispersal by wind and secondary dispersal by humans. These suggest that walking humans can disperse seeds to very long distances, up to at least 10 km, and provide some of the first quantified dispersal kernels for HMD.


Cell-connectivity-guided trajectory inference from single-cell data.
Single-cell RNA-sequencing enables cell-level investigation of cell differentiation, which can be modelled using trajectory inference methods. While tremendous effort has been put into designing these methods, inferring accurate trajectories automatically remains difficult. Therefore, the standard approach involves testing different trajectory inference methods and picking the trajectory giving the most biologically sensible model. As the default parameters are often suboptimal, their tuning requires methodological expertise. We introduce Totem, an open-source, easy-to-use R package designed to facilitate inference of tree-shaped trajectories from single-cell data. Totem generates a large number of clustering results, estimates their topologies as minimum spanning trees, and uses them to measure the connectivity of the cells. Besides automatic selection of an appropriate trajectory, cell connectivity enables to visually pinpoint branching points and milestones relevant to the trajectory. Furthermore, testing different trajectories with Totem is fast, easy, and does not require in-depth methodological knowledge. Totem is available as an R package at https://github.com/elolab/Totem.


The generalization of prior uncertainty during reaching.
Bayesian statistics defines how new information, given by a likelihood, should be combined with previously acquired information, given by a prior distribution. Many experiments have shown that humans make use of such priors in cognitive, perceptual, and motor tasks, but where do priors come from? As people never experience the same situation twice, they can only construct priors by generalizing from similar past experiences. Here we examine the generalization of priors over stochastic visuomotor perturbations in reaching experiments. In particular, we look into how the first two moments of the prior--the mean and variance (uncertainty)--generalize. We find that uncertainty appears to generalize differently from the mean of the prior, and an interesting asymmetry arises when the mean and the uncertainty are manipulated simultaneously.


Right TKR Patients Treated with Enhanced Pain and Rehabilitation Protocols Can Drive at 2 Weeks.
An important milestone in the recovery following total knee replacement (TKR) is the ability to return to driving. With advances in pain control and the widespread introduction of rapid rehab programs, we hypothesized that the ability to drive would also return sooner than had been traditionally observed. In our group of consecutive right TKR patients, using a driving simulator, we showed that at the 2-week mark, 36 of the 40 patients tested had returned to their preoperative driving capabilities and the other 4 had reached baseline at 3 weeks. While the eventual decision to return to driving is complex and dependent on many factors, we conclude that one of the benefits of enhanced pain and rehab protocols is that patients undergoing right TKR can return to driving in most instances at the 2-week mark rather than the traditional 6-week mark.


Genetic determinants of acute asthma therapy response in children with moderate-to-severe asthma exacerbations.
We documented inter-individual variability in the response to acute asthma therapy in children, attributed in part to five clinical factors (oxygen saturation, asthma severity score, virus detection, fever, symptoms between exacerbations; DOORWAY study). The contribution of genetic determinants of failure of acute asthma management have not been elucidated. We aim to determine single nucleotide polymorphisms (SNP) associated with emergency department (ED) management failure in children. A prospective cohort of 591 Caucasian children aged 1-17 years with moderate-to-severe asthma managed with standardized protocol were included. We examined 53 SNPs previously associated with asthma development, phenotypes, or bronchodilator or corticosteroids response. Associations between SNPs and management failure (hospitalization, active asthma management ≥8 h in ED, or a return visit within 72 h for one of two previous criteria) were examined using logistic regression, adjusting for the five clinical predictors of management failure. Four-hundred ninety-one subjects had complete clinical data and usable DNA samples. While controlling for clinical determinants, rs295137 in SPATS2L (OR = 1.77, 95%CI: 1.17, 2.68) was significantly associated with increased odds of ED management failure. Two SNPs in IL33 were associated with decreased odds of ED management failure: rs7037276 (OR = 0.55, 95%CI: 0.33, 0.90), and rs1342326 (OR = 0.52, 95%CI: 0.32, 0.86). The addition of these three SNPs to the clinical predictors significantly improved the model's predictive performance (P < 0.0004). Three SNPs were significantly associated with ED management failure in addition to clinical predictors, contributing to inter-individual variability. None has been previously associated with treatment response to acute asthma management.


The calcium-sensing receptor and insulin secretion: a role outside systemic control 15 years on.
In the 15 years since the identification and characterisation of the extracellular calcium-sensing receptor (CaR), it has become increasingly apparent that this cationic binding receptor is found in many tissues, not associated with the control of plasma calcium. One of these tissues is the pancreatic islet where insulin secretion provides the basis of energy regulation. It seems inherently unlikely that the islet responds to alterations in systemic calcium and a more plausible and intriguing possibility is that the CaR mediates cell-to-cell communication through local increases in the concentration of extracellular Ca(2+), co-released with insulin. This short article explores this possibility and suggests that this novel mechanism of cell communication, along with direct coupling via gap junctions and other local paracrine regulators helps explain why the glucose responsiveness of the intact islet is greater than the sum of the composite parts in isolation.


[Hand arthropathy of professional origin: milkingman hand].
The aim of this case-control study was to know possible correlates between distinctive pattern of hand arthrosis and length manual milking. Between January 1990 and January 1996, we retrospectively reviewed the patients with symptomatic arthrosis in whose hands was identified this pattern: degenerative disease of distal interphalangeal joints with radial deviation and flexion of distal phalanges in association to arthrosis involving metacarpophalangeal and interphalangeal joints of the thumb. The first 88 cases of hand arthrosis without this clinical and radiographic pattern make the control group. Patients suffering inflammatory rheumatic diseases were excluded. 35 patients with this special pattern of hand arthrosis were recruited. The majority of patients were women (94.3%) and coming from rural environment (94.3%). 32 patients had performed daily manual milking over a period of 20 years, but in three patients the pattern was not related to this activity (odds ratio = 928: p < 0.001). Symptoms were more frequent in the control-group (57% versus 26%; p < 0.04), in which no characteristic pattern of alignment abnormality was found. A typical pattern of hand arthrosis was associated with prolonged manual milking: degenerative disease of metacarpophalangeal and interphalangeal joints of thumb, radial deviation associated with flexion of distal phalanges of second at fourth fingers and arthrosis involving their distal interphalangeal joints, often linked to sight ulnar deviation of middle phalanges.


Single-molecule height measurements on microsomal cytochrome P450 in nanometer-scale phospholipid bilayer disks.
The architecture of membrane proteins in their native environment of the phospholipid bilayer is critical for understanding physiological function, but has been difficult to realize experimentally. In this communication we describe the incorporation of a membrane-anchored protein into a supported phospholipid bilayer. Cytochrome P450 2B4 solubilized and purified from the hepatic endoplasmic reticulum was incorporated into phospholipid bilayer nanostructures and oriented on a surface for visualization by atomic force microscopy. Individual P450 molecules were observed protruding from the bilayer surface. Problems associated with deformation of the protein by the atomic force microscopy probe were avoided by analyzing force-dependent height measurements to quantitate the height of the protein above the bilayer surface. Measurements of the atomic force microscopy cantilever deflection as a function of probe-sample separation reveal that the top of the P450 opposite the N-terminal membrane anchor region sits 3.5 nanometers above the phospholipid-water boundary. Models of the orientation of the enzyme are presented and discussed in relation to membrane interactions and interaction with cytochrome P450 reductase.


Short-Term CO2 Treatment of Harvested Grapes (Vitis vinifera L., cv. Trebbiano) before Partial Dehydration Affects Berry Secondary Metabolism and the Aromatic Profile of the Resulting Wine.
High CO2 concentrations applied to harvested horticultural products can modify primary and secondary metabolism. This work reports the metabolic responses to short-term CO2 treatments of white-skinned grapes (cv Trebbiano) undergoing postharvest partial dehydration. The influence of CO2 treatments on the aroma profile of the derived sweet wine was also assessed. Harvested grapes were treated with gaseous CO2 (30%) or air (control) for 24 h and then dehydrated (about 45% of weight loss) before vinification. Lipophilic and phenolic compounds of grape skin and the wine aroma profile were analyzed. In CO2-treated berries, the lipophilic and phenolic compounds decreased at a reduced and faster rate, respectively, during dehydration. Aroma profile of wine from CO2-treated grapes showed a slight but significantly higher content of glycosylated C13 and terpene compounds, and a decrease/absence of free acids, vanillin derivates and other phenol volatiles. The higher content of volatile alcohols in wine from treated berries suggests that the alcoholic fermentation was triggered. CO2 application before the withering process of Trebbiano grapes affects the aroma profile of the resulting wine by altering the free:glycosylated volatiles ratio. This study provides information on the possible use of CO2 as metabolic elicitor to modulate the aroma profile of the resulting wines obtained after grape dehydration.


Academic entitlement and academic performance in graduating pharmacy students.
To determine a measurable definition of academic entitlement, measure academic entitlement in graduating doctor of pharmacy (PharmD) students, and compare the academic performance between students identified as more or less academically entitled. Graduating students at a private health sciences institution were asked to complete an electronic survey instrument that included demographic data, academic performance, and 2 validated academic entitlement instruments. One hundred forty-one of 243 students completed the survey instrument. Fourteen (10%) students scored greater than the median total points possible on 1 or both of the academic entitlement instruments and were categorized as more academically entitled. Less academically entitled students required fewer reassessments and less remediation than more academically entitled students. The highest scoring academic entitlement items related to student perception of what professors should do for them. Graduating pharmacy students with lower levels of academic entitlement were more academically successful than more academically entitled students. Moving from an expert opinion approach to evidence-based decision-making in the area of academic entitlement will allow pharmacy educators to identify interventions that will decrease academic entitlement and increase academic success in pharmacy students.


Postprandial lipemia after short-term variation in dietary fat and carbohydrate.
Replacement of dietary fat with carbohydrate may not reduce the overall risk of coronary heart disease (CHD), because this elevates plasma triacylglycerol (TAG) concentrations. The lipoproteinemic effects of a high-carbohydrate diet are likely to be more marked shortly after the initiation of such a diet than after longer periods of intervention during which adaptive processes may counteract the initial effects. Therefore, we studied the postprandial responses to a standard meal after 3-day dietary intervention periods. An additional objective was to establish a model for future study of the mechanisms involved. Nine normolipidemic men consumed the meal (1.2 g fat, 1.1 g carbohydrate, and 0.2 g protein per 1 kg body mass) after 3 days on a high-carbohydrate diet (68% +/- 3% energy from carbohydrate, mean +/- SD) and also after 3 days on an isoenergetic high-fat diet (66% +/- 5% energy). Venous blood samples were obtained from fasted subjects and for 6 hours after the meal. In the fasted state, TAG was higher after the high-carbohydrate diet (1.18 +/- 0.18 v0.62 +/- 0.09 mmol/L, mean +/- SEM, P = .02) and high-density lipoprotein (HDL) cholesterol was lower (1.01 +/- 0.08 v 1.10 +/- 0.09 mmol/L, P = .002). The area under the plasma TAG concentration versus time curve was 42% +/- 7% higher after the high-carbohydrate diet (P = .003). After the high-carbohydrate diet, the postprandial insulin response did not differ between trials, but glucose and 3-hydroxybutyrate responses were lower (P = .009 and P = .02, respectively) and the lactate response was higher (P = .001). Plasma nonesterified fatty acids (NEFAs) were lower after the high-carbohydrate diet in the fasted state and for 4 hours postprandially, but were higher thereafter (interaction of time x trial, P = .001). These results indicate that compared with a high-fat diet, the plasma TAG response to a standard high-fat meal is markedly higher after a few days on a high-carbohydrate diet, with major differences in the associated metabolic milieu. The magnitude of these changes and the rapidity with which they developed suggest that this model may be attractive for future studies of the underlying mechanisms.


[Laterofacial malformations (maxillomandibular neurocristopathies) associated with anomalies of the brain stem and cranial nerves].
Neurological and cerebral computed tomography examinations were conducted in 25 infants and young children with uni- or bilateral maxillomandibular neurocristopathies (Goldenhar's, Franceschetti's first arch syndromes or transient forms). Taking all lesions into account, 18 children (82%) presented morphological and motor anomalies of the brain stem (mesencephalus, metencephalus) and its corresponding cranial nerves: four had morphological anomalies of the cerebellum and pons; four presented disorders of bulbopontine control (heart, lungs, deglutition); fifteen demonstrated sensory, sensorial, and motor deficiencies of the paired cranial nerves III, V, VI, VII, and VIII. The high frequency of ocular malformations in this series is emphasized: there were three cases of cryptophthalmia and one case of bilateral iris coloboma. These findings are in agreement with recent acquisitions in the field of cephalic developmental biology: the cephalic neural crest cells, after migration from the mesencephalorhombencephalic neural fold, contribute to the latero-facial and ocular organogenesis. The unity of origin of the face and brain provides an explanation for the predictive role of the former in case of malformations of the brain and its cranial nerves.


Validation of the American College of Foot and Ankle Surgeons Scoring Scales.
The American College of Foot and Ankle Surgeons (ACFAS) assembled a task force to develop a scoring scale that could be used by the membership and practitioners-at-large. The original publication that introduced the scale focused primarily on use of the scale and provided only brief background on the development of the health measurement instrument. Concerns regarding the validity and reliability of the scale were raised within the professional community, and ACFAS assembled a task force to address these concerns. The purpose of this article is to address the issues raised by reporting the detailed methods used in the development of the ACFAS Scoring Scales. The authors who constitute this task force reviewed the body of work previously conducted and applied standards that serve to evaluate the scoring scale for: 1) validity, 2) reliability, and 3) sensitivity to change. The results showed that a systematic and comprehensive approach was used in the development of the scoring scales, and the task force concluded that the statistical methods and instrument development process for all 4 modules of the scoring scales were conducted in an appropriate manner. Furthermore, modules 1 and 2 have been rigorously assessed and the elements of these modules have been shown to meet standards for validity, reliability, and sensitivity to change.


Cytogenetic analysis of the cSOD microregion in Drosophila melanogaster.
This report describes the genetic organization of a euchromatic region on the third chromosome of Drosophila melanogaster extending cytologically from 68A2 to C1, an interval comprising 10 or 11 polytene chromosome bands. The gene for cytoplasmic superoxide dismutase (cSOD) maps within this interval, as does low xanthine dehydrogenase (lxd).--Recessive lethal mutations were generated within the region by ethyl methanesulfonate mutagenesis and by hybrid dysgenesis. These lethals fall into 11 functional groups, which were partially ordered by complementation with deletions having breakpoints within the region. The distribution of dysgenesis-induced mutations in the region is highly nonrandom, the majority being within a single group. The mutability of this gene is comparable to that of singed (sn), a documented "hot-spot" for P-element insertion.--One of the EMS-induced lethals, l-108, fulfills biochemical criteria expected of a hypomorphic allele of cSOD. To our knowledge this is the first such allele recovered of this gene, and it should prove very useful in an analysis of the in vivo function of cytoplasmic SOD. Indeed, it has been demonstrated that cSOD is almost certainly a vital gene.


A syringe-like love dart injects male accessory gland products in a tropical hermaphrodite.
Sexual conflict shapes the evolution of many behaviours and processes involved in reproduction. Nearly all evidence supporting this comes from species where the sexes are separated. However, a substantial proportion of animals and most plants are hermaphroditic, and theoretical work predicts that sexual conflict plays an important role even when the sexes are joined within one individual. This seems to have resulted in bizarre mating systems, sophisticated sperm packaging and complex reproductive morphologies. By far the best-known example of such a strategy in hermaphrodites is the shooting of so-called love-darts in land snails. All known love darts carry a gland product on their outside and enter this into the partner's hemolymph by stabbing. Here, we show that species of the snail genus Everettia possess a syringe-like dart that serves as a real injection needle. Their dart is round in cross-section, contains numerous channels, and has perforations along its side. Histology and electron microscopy show that these holes connect to the channels inside the dart and run all the way up to the elaborate mucus glands that are attached to the dart sac. This is the first report on a love dart that is used as a syringe to directly inject the gland product into the partner's hemolymph. Although the exact use and function of this dart remains to be demonstrated, this clearly adds to the complexity of the evolution of reproductive strategies in hermaphrodites in general. Moreover, the perforations on the outside of the love dart resemble features of other injection devices, thus uncovering common design and repeated evolution of such features in animals.


Hepatoprotective effect of Opuntia microdasys (Lehm.) Pfeiff flowers against diabetes type II induced in rats.
Opuntia sp. has long been used as a folk medicine to treat hepatitis and diabetes in Sicile (Italy). To extract the polyphenols from the flower of Opuntia microdasys Lehm. at post flowring stage and evaluate the antidiabetic activity in vitro and in vivo. The hepatoprotective activity of Opuntia microdasys aqueous flowers extract at post flowering stage (OFP) has been tested for their antidiabetic activity. On fructose-alloxan induced diabete in rat model, evaluating the inhibitory effects of OFP on some carbohydrate metabolizing enzymes, pancreatic α-amylase and intestinal α-glucosidase activities in vitro. The OFP extract showed inhibitory activity against α-glucosidase (IC50=0.17±0.012mg/ml) and α-amylase (IC50=2.55±0.41mg/ml). The inhibitory potential of OFP extract on these enzymes suggests a positive and probable role of this extract in the management and treatment of diabetes mellitus, particularly, for type 2. Oral administration of the OFP at 200mg/kg to diabetic male rats for 28days demonstrated a significant protective effect by lowering the levels of glucose (123.21±1.38mg/dL) and hepatic marker enzymes (AST, ALT, LDH, γ-GT, BT, PAL, TC, LDL-C, HDL-C and TG). OFP attenuated oxidative stress by decreasing the SOD, CAT, GPX activity and the levels of PC and MDA in the liver and restored the histological architecture of the rat liver. OFP has protective effects on the protection of liver, thereby reducing some of the causes of diabetes in experimental animals.


Effects of linagliptin vs glimepiride on cognitive performance in type 2 diabetes: results of the randomised double-blind, active-controlled CAROLINA-COGNITION study.
Type 2 diabetes, particularly with concomitant CVD, is associated with an increased risk of cognitive impairment. We assessed the effect on accelerated cognitive decline (ACD) of the DPP-4 inhibitor linagliptin vs the sulfonylurea glimepiride in individuals with type 2 diabetes. The CAROLINA-COGNITION study was part of the randomised, double-blind, active-controlled CAROLINA trial that evaluated the cardiovascular safety of linagliptin vs glimepiride in individuals with age ≥40 and ≤85 years and HbA1c 48-69 mmol/mol (6.5-8.5%) receiving standard care, excluding insulin therapy. Participants were randomised 1:1 using an interactive telephone- and web-based system and treatment assignment was determined by a computer-generated random sequence with stratification by center. The primary cognitive outcome was occurrence of ACD at end of follow-up, defined as a regression-based index score ≤16th percentile on either the Mini-Mental State Examination (MMSE) or a composite measure of attention and executive functioning, in participants with a baseline MMSE score ≥24. Prespecified additional analyses included effects on ACD at week 160, in subgroups (sex, age, race, ethnicity, depressive symptoms, cardiovascular risk, duration of type 2 diabetes, albuminuria), and absolute changes in cognitive performance. Participants, caregivers, and people involved in measurements, examinations or adjudication, were all masked to treatment assignment. Of 6033 participants recruited from hospital and primary care sites, 3163 (38.0% female, mean age/diabetes duration 64/7.6 years, MMSE score 28.5, HbA1c 54 mmol/mol [7.1%]) represent the CAROLINA-COGNITION cohort. Over median 6.1 years, ACD occurred in 27.8% (449/1618, linagliptin) vs 27.6% (426/1545, glimepiride), OR 1.01 (95% CI 0.86, 1.18). Also, no differences in ACD were observed at week 160 (OR 1.07 [0.91, 1.25]), between treatments across subgroups, or for absolute cognitive changes. In a large, international outcome trial in people with relatively early type 2 diabetes at elevated cardiovascular risk, no difference in risk for ACD was observed between linagliptin and glimepiride over 6.1 years. This study was sponsored by Boehringer Ingelheim. ClinicalTrials.gov NCT01243424.


Discovery of the First Selective M4 Muscarinic Acetylcholine Receptor Antagonists with in Vivo Antiparkinsonian and Antidystonic Efficacy.
Nonselective antagonists of muscarinic acetylcholine receptors (mAChRs) that broadly inhibit all five mAChR subtypes provide an efficacious treatment for some movement disorders, including Parkinson's disease and dystonia. Despite their efficacy in these and other central nervous system disorders, antimuscarinic therapy has limited utility due to severe adverse effects that often limit their tolerability by patients. Recent advances in understanding the roles that each mAChR subtype plays in disease pathology suggest that highly selective ligands for individual subtypes may underlie the antiparkinsonian and antidystonic efficacy observed with the use of nonselective antimuscarinic therapeutics. Our recent work has indicated that the M4 muscarinic acetylcholine receptor has several important roles in opposing aberrant neurotransmitter release, intracellular signaling pathways, and brain circuits associated with movement disorders. This raises the possibility that selective antagonists of M4 may recapitulate the efficacy of nonselective antimuscarinic therapeutics and may decrease or eliminate the adverse effects associated with these drugs. However, this has not been directly tested due to lack of selective antagonists of M4. Here, we utilize genetic mAChR knockout animals in combination with nonselective mAChR antagonists to confirm that the M4 receptor activation is required for the locomotor-stimulating and antiparkinsonian efficacy in rodent models. We also report the synthesis, discovery, and characterization of the first-in-class selective M4 antagonists VU6013720, VU6021302, and VU6021625 and confirm that these optimized compounds have antiparkinsonian and antidystonic efficacy in pharmacological and genetic models of movement disorders.


Exposure to childhood infections and risk of Epstein-Barr virus--defined Hodgkin's lymphoma in women.
The role of Epstein-Barr virus (EBV) in Hodgkin's lymphoma (HL) etiology remains unresolved as EBV is detected in only some HL tumors and few studies have tried to reconcile its presence with factors suggesting viral etiology (e.g., childhood social class, infection history). In a population-based case-control study of San Francisco Bay area women, we analyzed interview data by tumor EBV status. Among 211 young adult cases, EBV-positive HL (11%) was associated with a single vs. shared bedroom at age 11 (OR = 4.0, 95% CI 1.1-14.4); risk was decreased for common childhood infections (OR = 0.3, 95% CI 0.1-1.0), including measles before age 10, but not with prior infectious mononucleosis (IM), which is delayed EBV infection. No study factors affected risk of young adult EBV-negative HL. Among 57 older adult cases, EBV-positive HL (23%) was unrelated to study factors; EBV-negative HL was associated with a single bedroom at age 11 (OR = 3.6, 95% CI 1.5-9.1) and IM in family members (OR = 3.1, 95% CI 1.1-9.0). Thus, delayed exposure to infection may increase risk of EBV-positive HL in young adults, but risk patterns differ in younger and older women for both EBV-positive and -negative HL. Late EBV infection does not appear relevant to risk, suggesting that other pathogens impact HL etiology in affluent female populations. Inconsistency of findings with prior studies may reflect failure of study risk factors to proxy meaningful exposures, risk differences by gender, or selection or misclassification bias. Null findings for EBV-negative HL indicate that etiologic models should be reconsidered for this common form.


Pulmonary function after bone marrow transplantation for chronic myeloid leukaemia.
Pulmonary function was measured before and at intervals after treatment in 44 patients who received a bone marrow transplant for chronic myeloid leukaemia in the chronic phase. All patients were treated with cytotoxic drugs, total body irradiation, and post-graft immunosuppression. Thirty four patients surviving for 12 months were followed at three monthly intervals and 16 patients for 24 months. Fifteen patients received unmanipulated donor marrow cells and 29 patients received donor marrow cells depleted of lymphocytes ex vivo with the monoclonal antibody Campath-1. The 21 patients treated early in this study received 10 Gy of total body irradiation whereas the 23 patients treated more recently, who were all T lymphocyte depleted, received 12 Gy. Pretransplant lung function for the group was normal and was similar in survivors (n = 34) and nonsurvivors (n = 10), and in smokers (n = 8) and non-smokers (n = 36). (Carbon monoxide transfer factor--TLCO) was under 75% of predicted normal in nine patients before transplantation. TLCO, carbon monoxide transfer coefficient (KCO), FEV1, and vital capacity (VC) values were lower 6 and 12 months after bone marrow transplant than initially. The greatest decline was in TLCO, from an initial value of 89% to 66% at 6 and 70% at 12 months. The 16 longer term survivors showed significant recovery of function between 6 and 24 months after bone marrow transplant for TLCO, KCO, and VC, the increase ranging from 6.3% to 7.3% predicted. Airflow obstruction (FEV1/VC ratio less than 70%) developed in one patient. The major factors associated with deterioration in pulmonary function at 6 and 12 months after transplantation in the 34 survivors (stepwise multiple regression analysis) were (a) transplantation with T cell depleted donor marrow (p less than 0.005) and higher total body irradiation dose (p less than 0.02) with a fall in KCO and an increase in the FEV1/VC ratio; (b) chronic graft versus host disease with a fall in VC (p less than 0.01); and less fall in KCO (p less than 0.01); and (c) acute graft versus host disease with a fall in FEV1 (p less than 0.01). It is considered that most patients who survive the short term risks of bone marrow transplant have only minor long term impairment of pulmonary function.


[Suppressive Effects of Extract of Cedar Wood on Heat-induced Expression of Cellular Heat Shock Protein].
In recent years, highly antimicrobial properties of cedar heartwood essential oil against the wood-rotting fungi and pathogenic fungi have been reported in several papers. Antimicrobial properties against oral bacteria by hinokitiol contained in Thujopsis have been also extensively studied. The relation of naturally derived components and human immune system has been studied in some previous papers. In the present study, we focused on Japanese cedar, which has the widest artificial afforestation site in the country among various tree species. Extract oil was obtained from mixture of sapwood and heartwood of about 40-year cedar grown in Oguni, Kumamoto, Japan. We examined the influence of extract components from Japanese cedar woods on the expression of heat shock protein 70 (Hsp70) during heating, and on the micronucleus formation induced by the treatment of bleomycin as a DNA damaging agent. Cell lines used in this study were human fetal glial cells (SVGp12) and human glioma cells (MO54). Remarkable suppression of the Hsp70 expression induced by heating at 43°C was detected by the treatment of cedar extract in both SVGp12 and MO54 cells. We also found that cedar extract had an inhibitory tendency to reduce the micronucleus formation induced by bleomycin. From these results, the extract components from Japanese cedar woods would have an inhibitory effect of the stress response as a suppression of the heat-induced Hsp70 expression, and might have a reductive effect on carcinogenicity.


Femoral structure and stiffness in patients with femoral neck fracture.
Bone morphological characteristics may relate to the risk of hip fracture. We applied finite element modeling to radiologic data for two groups of women in vivo to address two questions: (a) Do individuals who have just sustained a femoral neck fracture exhibit reduced three-dimensional structural stiffness? and (b) Are victims of hip fracture disproportionately more susceptible to loads sustained in a fall than to stance-type loads? Ten white women (age: 64-76 years) who had just sustained a femoral neck fracture and 18 female volunteers (age: 65-76 years), matched as groups for race, age. and body mass index, were evaluated. From quantitative computed tomography scans, femoral morphometric and volumetric cancellous density measurements were obtained and a finite element model was constructed. Two load conditions were simulated: single-stance phase and lateral fall. Global stiffness values were determined for each model. The cancellous bone density was significantly lower at the femoral neck and the femoral neck and head diameters were significantly larger in the women in the fracture group than in those in the control group. The stiffness of the proximal femur did not differ significantly between the groups for either load condition. An apparently linear relationship was found for stiffness at stance load compared with stiffness at fall load (r = 0.84, p < 0.001). and slopes did not differ significantly between the groups. Although cancellous density was reduced at the fracture site in patients with femoral neck fractures. this did not result in a reduction in the predicted bone stiffness. Previous studies have established a very strong relationship between the stiffness and strength of bone. Since these modeling methods were thoroughly validated ex vivo, we conclude that although decreased bone density at the femoral neck may predict where fracture initiates, the risk of hip fracture per se may be more strongly dependent on issues such as the risk of falling and fall biomechanics than on the structural characteristics of bone.


Adipocyte inflammation is essential for healthy adipose tissue expansion and remodeling.
Chronic inflammation constitutes an important link between obesity and its pathophysiological sequelae. In contrast to the belief that inflammatory signals exert a fundamentally negative impact on metabolism, we show that proinflammatory signaling in the adipocyte is in fact required for proper adipose tissue remodeling and expansion. Three mouse models with an adipose tissue-specific reduction in proinflammatory potential were generated that display a reduced capacity for adipogenesis in vivo, while the differentiation potential is unaltered in vitro. Upon high-fat-diet exposure, the expansion of visceral adipose tissue is prominently affected. This is associated with decreased intestinal barrier function, increased hepatic steatosis, and metabolic dysfunction. An impaired local proinflammatory response in the adipocyte leads to increased ectopic lipid accumulation, glucose intolerance, and systemic inflammation. Adipose tissue inflammation is therefore an adaptive response that enables safe storage of excess nutrients and contributes to a visceral depot barrier that effectively filters gut-derived endotoxin.


Asymmetries of the balanced SSFP profile. Part I: theory and observation.
The signal in balanced steady-state free precession has a strong sensitivity to off-resonance, which is typically described in terms of a signal "profile" over a range of frequencies. This profile has a well-known form for homogeneous media with a single T(1), T(2), and resonance frequency, which is symmetric about the on-resonance frequency. However, a straightforward extension to this established signal model predicts that the profile may become asymmetric in the presence of inhomogeneous frequency content, as would be expected to happen in tissue due to microstructural boundaries, compartments, and chemical shift. The presence of asymmetries in the balanced steady-state free precession profile may therefore provide a marker of tissue integrity. This manuscript describes the theory behind balanced steady-state free precession asymmetries, a method for detecting these effects, and the first measurements of balanced steady-state free precession asymmetries in tissue. Asymmetries are found in gray matter, white matter, and muscle, with excellent reproducibility. A companion paper considers the large white matter asymmetries in more detail.


Use of an Internet portal to improve community-based pediatric ADHD care: a cluster randomized trial.
To determine the effectiveness of a quality improvement program to improve pediatricians' adherence to existing, evidence-based, attention-deficit/hyperactivity disorder (ADHD) practice guidelines. Forty-nine community-based pediatricians at 8 practices participated in a cluster-randomized trial. Practices were matched according to the numbers of pediatricians and the proportions of patients receiving Medicaid. The medical charts for a random sample of patients with ADHD for each of the participating pediatricians were examined at baseline and 6 months. All practices participated in 4 sessions of training, including didactic lectures and office flow modification workshops. Practices were then given access to an ADHD Internet portal that allowed parents, teachers, and pediatricians to input information (eg, rating scales) about patients, after which information was scored, interpreted, and formatted in a report style that was helpful for assessment and treatment of patients with ADHD. Physicians evaluated their practice behaviors quarterly and addressed underperforming areas. Pediatricians in the intervention group, compared with those in the control group, demonstrated significantly higher rates of many American Academy of Pediatrics-recommended ADHD care practices, including collection of parent (Cohen's d = 0.69) and teacher (d = 0.68) rating scales for assessment of children with ADHD, use of Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, criteria (d = 0.85), and use of teacher rating scales to monitor treatment responses (d = 1.01). A quality improvement intervention that can be widely disseminated by using Internet-based information technology significantly improved the quality of ADHD care in community-based pediatric settings.


Anti-Coronavirus Disease 2019 Vaccines: Need for Informed Consent.
Vaccines are among some of the most efficacious medical and public health methods ever employed to contain a pandemic, in addition to providing protective and preventive measures. Evaluation of vaccineassociated adverse events through experimentation and empirical evidence is an integral part ofthoroughly assessing the safety of vaccines before authorization of their widespread use. History has highlighted the importance of continuous search for possible vaccine-related adverse effects and vaccine-induced immunogenicity long afterlicensure, suggesting that a primary concern with new vaccines is not only efficacy but also safety, particularly overthe long term. Many of the various anti-COVID-19 vaccines have used different types of technology, with some being introduced for the first time or rushed shortly into testing, bypassing animal experimentations. They have been adopted for use through emergency use authorizations, leading to a less than optimal collection of broad data on safety, immunogenicity, effectiveness, and time span of protection, as well as short follow-up of few months, despite many infectious disease experts arguing thatit takes 10 years to develop a vaccine. Given the valid concerns on well-recognized short-term and long-term safety issues, such as antibody-dependent enhancement and other processes like molecular mimicry and potential genomic transformation, the experimental nature of the vaccination process, the limited shortterm follow-up in the main trials, and the dismissal by law of pharma companies and health care providers from any medico-legal responsibilities, the application of an informed consent should become not only a necessity but also mandatory by law in accordance with all declarations on human rights. Such information should be provided to every potential recipient in the form of an official written digital consent prior to the registration for or the receipt of the vaccine.


A comparison of the EU regulatory approach to directed mutagenesis with that of other jurisdictions, consequences for international trade and potential steps forward.
A special regulatory regime applies to products of recombinant nucleic acid modifications. A ruling from the European Court of Justice has interpreted this regulatory regime in a way that it also applies to emerging mutagenesis techniques. Elsewhere regulatory progress is also ongoing. In 2015, Argentina launched a regulatory framework, followed by Chile in 2017 and recently Brazil and Colombia. In March 2018, the USDA announced that it will not regulate genome-edited plants differently if they could have also been developed through traditional breeding. Canada has an altogether different approach with their Plants with Novel Traits regulations. Australia is currently reviewing its Gene Technology Act. This article illustrates the deviation of the European Union's (EU's) approach from the one of most of the other countries studied here. Whereas the EU does not implement a case-by-case approach, this approach is taken by several other jurisdictions. Also, the EU court ruling adheres to a process-based approach while most other countries have a stronger emphasis on the regulation of the resulting product. It is concluded that, unless a functioning identity preservation system for products of directed mutagenesis can be established, the deviation results in a risk of asynchronous approvals and disruptions in international trade.


The effect of extended family living on the mental health of three generations within two Asian communities.
A study by Shah & Sonuga-Barke (1995) identified a relationship between family structure and the mental health of Pakistani Muslim mothers and their children. Children in extended families fared better, but their mothers fared worse than their nuclear family counterparts. The present study replicates and extends this study by exploring the impact of nuclear and extended family living on the mental health of three generations (children, mothers and grandmothers) in British Hindu as well as Muslim communities. 44 Muslim and 42 Hindu families participated in the study. The mental health of mothers and grandmothers and the behavioural problems of children (aged 5-11) were examined. Both mothers and grandmothers completed the Hospital Anxiety and Depression Scale. The children's behavioural adjustment was rated by their teachers using the Rutter Scale. Other relevant variables such as acculturation levels were also measured. Children and grandmothers were better adjusted in extended families than nuclear families. In contrast, mothers were better adjusted in nuclear families. This interaction between family type and generation was evident in both Muslim and Hindu families and did not appear to be mediated by other variables such as acculturation. Furthermore, mothers' and childrens' adjustment was significantly correlated with grandmothers', but not mothers', mental health in extended families (although not in nuclear families). These results provide further evidence for the link between family structure and mental health in Asian communities. They also challenge some of the assumptions about maternal mental health, its effects on child adjustment and its links to systems of social support. In extended families where social support was likely to be most available mothers were at greatest risk, while their children profited and this advantage seemed to be linked to the grandmaternal presence.


Comparison of aspirin and benorylate in the treatment of rheumatoid arthritis.
In a double-blind between-patient study of aspirin and benorylate carried out in 72 outpatients with rheumatoid arthritis, benorylate 4 g twice daily was shown to be an effective analgesic and anti-inflammatory drug, its effects being indistinguishable from those of aspirin 1.2 g four times daily. Compared with the pretreatment values both drugs produced a statistically significant improvement (P < 0.01) in functional grade, overall pain, articular index, and grip strength at the end of the first and second weeks. The overall incidence of side effects was less with benorylate, though this difference was not significant at the 5% level.


Polyacetylenes from Oplopanax horridus and Panax ginseng: Relationship between Structure and PPARγ Activation.
Oplopanax horridus and Panax ginseng are members of the plant family Araliaceae, which is rich in structurally diverse polyacetylenes. In this work, we isolated and determined structures of 23 aliphatic C17 and C18 polyacetylenes, of which five are new compounds. Polyacetylenes have a suitable scaffold for binding to PPARγ, a ligand-activated transcription factor involved in metabolic regulation. Using a reporter gene assay, their potential was investigated to activate PPARγ. The majority of the polyacetylenes showed at least some PPARγ activity, among which oplopantriol B 18-acetate (1) and oplopantriol B (2) were the most potent partial PPARγ activators. By employing in silico molecular docking and comparing the activities of structural analogues, features are described that are involved in PPARγ activation, as well as in cytotoxicity. It was found that the type of C-1 to C-2 bond, the polarity of the terminal alkyl chain, and the backbone flexibility can impact bioactivity of polyacetylenes, while diol structures with a C-1 to C-2 double bond showed enhanced cytotoxicity. Since PPARγ activators have antidiabetic and anti-inflammatory properties, the present results may help explain some of the beneficial effects observed in the traditional use of O. horridus extracts. Additionally, they might guide the polyacetylene-based design of future PPARγ partial agonists.


Beetroot juice is more beneficial than sodium nitrate for attenuating muscle pain after strenuous eccentric-bias exercise.
The aim of this study was to compare the effects of beetroot juice (BTJ) and a nitrate only drink (sodium nitrate; SN) on indices of exercise-induced muscle damage (EIMD). Thirty recreationally active males consumed either BTJ (n = 10), a nitrate-matched SN drink (n = 10), or an isocaloric placebo (PLA; n = 10) immediately and at 24 and 48 h after performing 100 drop jumps. To assess muscle damage, maximal isometric voluntary contractions (MIVCs), countermovement jumps (CMJs), pressure-pain threshold (PPT), creatine kinase (CK), and high-sensitivity C-reactive protein (hs-CRP) were measured before, immediately after and at 24, 48, and 72 h following the drop jumps. BTJ and SN increased serum nitric oxide, which peaked at 2 h post-ingestion (136 ± 78 and 189 ± 79 μmol·L-1, respectively). PPT decreased in all groups postexercise (P = 0.001), but was attenuated with BTJ compared with SN and PLA (P = 0.043). PPT was 104% ± 26% of baseline values at 72 h after BTJ, 94% ± 16% after SN, and 91% ± 19% after PLA. MIVC and CMJ were reduced following exercise (-15% to 25%) and did not recover to baseline by 72 h in all groups; however, no group differences were observed (P > 0.05). Serum CK increased after exercise but no group differences were present (P > 0.05). hsCRP levels were unaltered by the exercise protocol (P > 0.05). These data suggest that BTJ supplementation is more effective than SN for attenuating muscle pain associated with EIMD, and that any analgesic effects are likely due to phytonutrients in BTJ other than nitrate, or interactions between them.


Exercise in the Treatment of Addiction: A Systematic Literature Review.
Exercise has been recognized as a promising and emerging treatment for individuals recovering from addiction. The purpose of this article was to systematically review scientific studies using exercise as a means to improve, sustain, or treat addictions, and to provide suggestions for the future use of exercise as a treatment method for addiction. Using PRISMA guidelines, a database search was conducted for articles that tested the impact of exercise interventions on addiction-related outcomes. To be included, peer-reviewed experimental design studies had to use human subjects to investigate the relationship between exercise and the treatment of or recovery from addiction. Garrard's Matrix Method was used to extract data from reviewed articles (n = 53). Nearly three quarters of the studies reviewed documented a significant change in addiction-related outcomes (e.g., more days abstinent, reduced cravings) in response to exercise exposure, particularly while someone was receiving treatment at an in or outpatient clinic. Many studies investigated the effect of acute bouts of exercise on nicotine dependence, and many studies had small sample sizes, leaving room for future research on how exercise might benefit people recovering from substance and process addictions. Results affirm that exercise can be a helpful aspect of addiction treatment. Future researchers should investigate different exercise settings (e.g., group-based exercise vs individual) and explore exercise maintenance and the long-term outcomes following discharge from treatment facilities.


Resonance of cortico-cortical connections of the motor system with the observation of goal directed grasping movements.
Goal directed movements require the activation of parietal, premotor and primary motor areas. In monkeys, neurons of these areas become active also during the observation of movements performed by others, especially for coding the goal of the action (mirror system). Using bifocal transcranial magnetic stimulation (TMS) in healthy subjects, we tested whether the observation of goal directed reach to grasp actions may lead to specific changes in the short-latency connections linking key areas of the mirror system, such as the anterior intraparietal cortex (AIP) and the ventral premotor cortex (PMv), with the primary motor cortex (M1). We found that AIP-M1 and PMv-M1 cortico-cortical interactions were specifically activated when observing successful reaching to grasp goal directed actions, in which the hand posture was congruent with the goal of the action performed by the actor. On the other hand they were not modified when the same goal directed actions were performed wrongly with an inappropriate grasping posture. A similar profile of excitability was observed when testing specific intracortical facilitatory circuits in M1 (I(2)-waves), known to reflect the activity in cortico-cortical pathways transmitting information from PMv. We conclude that the simple observation of others' goal directed actions is able to induce specific neurophysiological changes in some cortico-cortical circuits of the human motor system.


Atrial flutter in the fetus and young infant: an association with accessory connections.
To highlight the association between atrial flutter and accessory connections in the fetus and young infant. A retrospective review from January 1985 to January 1990. Fetuses, neonates, and young infants with atrial flutter. Four fetuses and five infants presented with atrial flutter. In two fetuses and one infant sinus rhythm returned spontaneously. The other six required cardioversion. Three of them developed orthodromic atrioventricular re-entry tachycardia and each had evidence of an accessory connection. Because atrial flutter in the fetus and neonate is rare, the high incidence of accessory connections in this group points to a possible aetiology of "idiopathic" atrial flutter in this age group.


p53, a target of estrogen receptor (ER) α, modulates DNA damage-induced growth suppression in ER-positive breast cancer cells.
In response to genotoxic stress, the p53 tumor suppressor induces target genes for cell cycle arrest, apoptosis, and DNA repair. Although p53 is the most commonly mutated gene in all human cancers, it is only mutated in about 20% of breast cancers. 70% of all breast cancer cases are estrogen receptor (ER)-positive and express ERα. ER-positive breast cancer generally indicates good patient prognosis and treatment responsiveness with antiestrogens, such as tamoxifen. However, ER-positive breast cancer patients can experience loss or a reduction in ERα, which is associated with aggressive tumor growth, increased invasiveness, poor prognosis, and loss of p53 function. Consistent with this, we found that p53 is a target gene of ERα. Specifically, we found that knockdown of ERα decreases expression of p53 and its downstream targets, MDM2 and p21. In addition, we found that ERα activates p53 transcription via binding to estrogen response element half-sites within the p53 promoter. Moreover, we found that loss of ERα desensitizes, whereas ectopic expression of ERα sensitizes, breast cancer cells to DNA damage-induced growth suppression in a p53-dependent manner. Altogether, this study provides an insight into a feedback loop between ERα and p53 and a biological role of p53 in the DNA damage response in ER-positive breast cancers.


The influence of substituting dietary peptide-bound with free amino acids on nitrogen metabolism and acid-base balance of broiler chickens depends on asparagine and glutamine supply.
Reducing dietary crude protein (CP) concentration while maintaining adequate amino acid (AA) supply by free AA inclusion can contribute to attenuate the negative environmental effects of animal farming. This study investigated upper limits of dietary free AA inclusions without undesirable effects including the dependence on asparagine (Asn) and glutamine (Gln) supply. Ten broilers were allocated to sixty-three metabolism units each and offered nine experimental diets from day (d) 7-21 (n 7). One diet (167 g CP/kg) contained 80 g soya protein isolate (SPI)/kg. In the other diets, 25, 50, 75 and 100 % of the digestible AA from SPI were substituted with free AA. Digestible Asn+aspartic acid (Asp) and Gln+glutamic acid (Glu) were substituted with Asp/Glu or 50/50 mixes of Asp/Asn and Glu/Gln, respectively. Total excreta were collected from d 11-14 and from d 18-21. Growth and nitrogen accretion were unaffected by 25 and 50 % substitution without and with free Asn/Gln, respectively, but decreased at higher substitution (P ≤ 0·024). Circulating concentrations of Asp, Glu and Gln were unaffected by treatment, while Asn decreased at substitution higher than 50 % when Asn/Gln were not provided (P ≤ 0·005). Blood gas analysis on d 21 indicated a compensated metabolic acidosis at substitution higher than 50 and 75 % without and with free Asn/Gln, respectively (P ≤ 0·017). Results suggest that adding Asn/Gln increased an upper limit for proportion of dietary free AA from 10 to 19 % of dietary CP and enabled higher free AA inclusion without affecting the acid-base balance.


Unusual peaks and baseline shifts in capillary electrophoresis.
Emersion peaks, baseline shifts, and spontaneous-marker peaks in CE are documented. These baseline perturbations are observed in a simple CE system that does not employ sample injection and with sodium benzoate as the running electrolyte. Emersion peaks are attributed to the preferential adsorption of electrolyte at the air-solution interface formed upon emersion of the capillary inlet. Baseline shifts are believed to be caused by field-dependent changes in electrolyte concentration entering the capillary. These shifts may be accompanied by spontaneous-marker peaks, which may be caused by diffusion into or out of the capillary inlet between electrophoretic experiments. All baseline perturbations are detected as a change in UV absorbance signal and are transported by electroosmotic flow.


α-Diketone formation accompanied by oxidation of sulfur functional group by the reaction of o-alkynylarenesulfoxide with iodine.
The reaction of o-alkynylarenesulfoxide with iodine was investigated in detail, revealing functionalities of the formation of α-diketones with sulfenyl, sulfinyl, and sulfonyl. Additives can change the ratio of products to give medium-to-excellent yields. Results show that water is taken into only sulfonyl compound and that other oxygen atoms constructed in the products are presumably derived from sulfoxide of the starting material and molecular oxygen.


Loneliness and the Social Brain: How Perceived Social Isolation Impairs Human Interactions.
Loneliness is a painful condition associated with increased risk for premature mortality. The formation of new, positive social relationships can alleviate feelings of loneliness, but requires rapid trustworthiness decisions during initial encounters and it is still unclear how loneliness hinders interpersonal trust. Here, a multimodal approach including behavioral, psychophysiological, hormonal, and neuroimaging measurements is used to probe a trust-based mechanism underlying impaired social interactions in loneliness. Pre-stratified healthy individuals with high loneliness scores (n = 42 out of a screened sample of 3678 adults) show reduced oxytocinergic and affective responsiveness to a positive conversation, report less interpersonal trust, and prefer larger social distances compared to controls (n = 40). Moreover, lonely individuals are rated as less trustworthy compared to controls and identified by the blinded confederate better than chance. During initial trust decisions, lonely individuals exhibit attenuated limbic and striatal activation and blunted functional connectivity between the anterior insula and occipitoparietal regions, which correlates with the diminished affective responsiveness to the positive social interaction. This neural response pattern is not mediated by loneliness-associated psychological symptoms. Thus, the results indicate compromised integration of trust-related information as a shared neurobiological component in loneliness, yielding a reciprocally reinforced trust bias in social dyads.


Targeting dsRNA-specific single-chain Fv antibody fragments to different cellular locations in Nicotiana tabacum L.
Expression of antibodies or antibody fragments in plants is a useful tool for producing active antibody derivatives for diagnostic or pharmaceutical purposes as well as for immunomodulation. We investigated the effect of cellular expression site on the stability and yield of double-stranded RNA (dsRNA)-specific single-chain Fv-fragments (scFv) in transgenic tobacco. Two antibodies (J2 and P6) belonging to the V23(J558) heavy chain variable gene family but differing in the light chain variable domain were used. scFvs were targeted to the cytoplasm - with or without anchoring them in the plasma membrane -, into the endoplasmic reticulum (ER) and to the apoplast. Although high mRNA concentrations were detected in all cases, scFv proteins accumulated only when scFvs were made ER-resident by appropriate signal sequences. When the ER retention signal was removed to allow scFv-secretion to the apoplast, no scFv-proteins were detected. Despite the strong homology of the VH-sequences of J2 and P6 antibodies, only P6 provided a stable scFv scaffold for intracytoplasmic expression. J2-scFv could not be stabilised either by adding a C-terminal stabilisation signal or by anchoring the protein on the cytoplasmic side of the plasma membrane (PM). It was found that dsRNA-specific J2-scFvs are active in vivo and enhance Potato Virus Y induced symptoms in infected tobacco. This is the first report describing the expression and biological effect of RNA-specific antibodies in plants.


Validation of Indian Council of Medical Research Neurocognitive Tool Box in Diagnosis of Mild Cognitive Impairment in India: Lessons from a Harmonization Process in a Linguistically Diverse Society.
In a linguistically diverse country such as India, challenges remain with regard to diagnosis of early cognitive decline among the elderly, with no prior attempts made to simultaneously validate a comprehensive battery of tests across domains in multiple languages. This study aimed to determine the utility of the Indian Council of Medical Research-Neurocognitive Tool Box (ICMR-NCTB) in the diagnosis of mild cognitive impairment (MCI) and its vascular subtype (VaMCI) in 5 Indian languages. Literate subjects from 5 centers across the country were recruited using a uniform process, and all subjects were classified based on clinical evaluations and a gold standard test protocol into normal cognition, MCI, and VaMCI. Following adaptation and harmonization of the ICMR-NCTB across 5 different Indian languages into a composite Z score, its test performance against standards, including sensitivity and specificity of the instrument as well as of its subcomponents in diagnosis of MCI, was evaluated in age and education unmatched and matched groups. Variability in sensitivity-specificity estimates was noted between languages when a total of 991 controls and 205 patients with MCI (157 MCI and 48 VaMCI) were compared due to a significant impact of age, education, and language. Data from a total of 506 controls, 144 patients with MCI, and 46 patients with VaMCI who were age- and education-matched were compared. Post hoc analysis after correction for multiple comparisons revealed better performance in controls relative to all-cause MCI. An optimum composite Z-score of -0.541 achieved a sensitivity of 81.1% and a specificity of 88.8% for diagnosis of all-cause MCI, with a high specificity for diagnosis of VaMCI. Using combinations of multiple-domain 2 test subcomponents retained a sensitivity and specificity of >80% for diagnosis of MCI. The ICMR-NCTB is a "first of its kind" approach at harmonizing neuropsychological tests across 5 Indian languages for the diagnosis of MCI due to vascular and other etiologies. Utilizing multiple-domain subcomponents also retains the validity of this instrument, making it a valuable tool in MCI research in multilingual settings.


Dyadic parent-child interaction during early childhood: contributions of parental and child personality traits.
Individual differences in personality play a major role for functioning in a multitude of important life domains, including one's interpersonal relationships. The present study examined the effects of parental personality and child temperament traits on dyadic parent-child interaction during early childhood. Participants were a community sample of 145 mothers, 145 fathers, and their 3- to 6-year-old children. Child traits were assessed using standardized laboratory paradigms, parents reported on their own traits, and parent-child interaction was assessed observationally. Parental positive emotionality, negative emotionality, and constraint subtraits were related to parental responsiveness; the number and type of parental bids and the quality of parental responsiveness were also a function of child positive emotionality and effortful control subtraits and, for mothers, child negative emotionality subtraits. Child traits were related to their own interaction behaviors; children higher on positive emotionality subtraits made more social bids, and children higher on effortful control subtraits made more influence attempts and fewer negative bids; child positive emotionality and effortful control subtraits were associated with higher quality child responsiveness. Findings speak to coherence in personality constructs across the life span, with comparable traits measured in adults and early childhood-aged children demonstrating remarkably consistent effects on dyadic interaction behavior.


Antigenic modulation of measles subacute sclerosing panencephalitis virus in a persistently infected rat glioma cell line by monoclonal anti-haemagglutinin antibodies.
Rat glioma C6 cells persistently infected with measles subacute sclerosing panencephalitis (SSPE) virus (C6/SSPE) express the viral membrane proteins haemagglutinin (HA) and F on their cell surface as well as the intracellular proteins N, P and M. Previously we have shown that the addition of a polyclonal antibody against the HA antigen to the growth medium of C6/SSPE cells led to a gradual loss of all viral antigens. Here we show that the addition of a monoclonal antibody (MAb K83) leads only to a transient decrease in viral antigens during the first three passages. After the third passage viral antigens start to increase and after five passages they produce more antigens than at the premodulation level. At this point of the MAb treatment, MAb K83 no longer recognized the HA antigen on the surface of the cells and in virus particles produced by these cells in contrast with polyclonal antibodies or other MAbs against the HA antigen. The results suggest that specific variants of the SSPE virus with an altered HA antigen were selected by the MAb treatment.


Long-acting beta 2-adrenoceptor agonists: a new perspective in the treatment of asthma.
New long-acting beta 2-adrenoceptor agonists, formoterol and salmeterol, may soon appear in several European countries for treatment of asthma. This review examines currently available information and compares the basic pharmacology and describes the clinical effects of these new drugs. The long duration of bronchodilation seen in clinical studies seems to be similar, whereas in isolated tissues there might be a difference in the binding characteristics to the beta 2-adrenoceptor. Long-acting beta 2-agonists could have an inhibitory effect on inflammatory events related to asthma, but the clinical relevance of these effects is not clear at present. Long-term studies up to one year with both new drugs have not shown any unexpected side-effects, and no tachyphylaxis to beta-adrenoceptor stimulation has been reported. Patients appear to strongly prefer the new drugs compared to the short-acting beta 2-agonists. The potential place for these drugs in the treatment of asthma is discussed and some pitfalls pointed out. It is likely that the long-acting beta 2-agonists will be beneficial to many asthmatic patients.