Higher numbers of memory B-cells and Th2-cytokine skewing in high responders to hepatitis B vaccination.
In the present study, differences in hepatitis B surface antigen (HBsAg)-specific memory B-cell responses between low and high responders to hepatitis B vaccine (HepB), based on levels of antibodies against HBsAg (anti-HBs), were determined. In addition, HBsAg specific T-cell responses between high (anti-HBs level >20,000 IU/L) and low (anti-HBs level <1500 IU/L) responders were compared. Numbers of HBsAg-specific B-cells, plasma immunoglobulin G (Ig) levels, and T-cell cytokine concentrations were measured in low and high responders directly before and one month after the second booster vaccination. In advance, an Enzyme-linked Immunosorbent Spot (ELISpot) Assay was optimized for the determination of HBsAg-specific B-cell responses. The number of HBsAg-specific B-cells was significantly higher (p<0.01) in the high responder group compared to the low responder group after a booster vaccination with HepB. In addition, the plasma IgG levels and numbers of HBsAg-specific B-cells were significantly correlated (RS=0.66, p<0.01). The HBsAg-specific Th1 cell response showed the same values in the low and high responder group and did not change by the booster vaccination with HepB. However, a significant correlation (RS=0.6975, p=0.007) between the IL-13 levels and the plasma IgG levels post-booster was found. Subsequently, the IL-13 level in the high-responder group post-booster was significantly higher compared to the low-responder group. Since activation of the B-cell response after vaccination is induced by Th2 cells and IL-13 is produced by these cells, we conclude that the difference in HBsAg-specific Th2 cells is involved in determining the differences in anti-HBs level and memory B-cell numbers between low and high responders.


Propagule pressure and colony social organization are associated with the successful invasion and rapid range expansion of fire ants in China.
We characterized patterns of genetic variation in populations of the fire ant Solenopsis invicta in China using mitochondrial DNA sequences and nuclear microsatellite loci to test predictions as to how propagule pressure and subsequent dispersal following establishment jointly shape the invasion success of this ant in this recently invaded area. Fire ants in Wuchuan (Guangdong Province) are genetically differentiated from those found in other large infested areas of China. The immediate source of ants in Wuchuan appears to be somewhere near Texas, which ranks first among the southern USA infested states in the exportation of goods to China. Most colonies from spatially distant, outlying areas in China are genetically similar to one another and appear to share a common source (Wuchuan, Guangdong Province), suggesting that long-distance jump dispersal has been a prevalent means of recent spread of fire ants in China. Furthermore, most colonies at outlier sites are of the polygyne social form (featuring multiple egg-laying queens per nest), reinforcing the important role of this social form in the successful invasion of new areas and subsequent range expansion following invasion. Several analyses consistently revealed characteristic signatures of genetic bottlenecks for S. invicta populations in China. The results of this study highlight the invasive potential of this pest ant, suggest that the magnitude of international trade may serve as a predictor of propagule pressure and indicate that rates and patterns of subsequent range expansion are partly determined by the interplay between species traits and the trade and transportation networks.


Haemodynamics in children during rest and exercise: methods and normal values.
Measuring haemodynamic performance in children is either invasive, and thus unacceptable, or noninvasive when the measured variable is often remote from the true variable. Measuring only maximum performance variables relies too heavily on motivation, especially in disease groups. We describe a method for the measurement of haemodynamic performance using respiratory mass spectrometry during rest, exercise and recovery therefrom. One hundred and six healthy children (55 male, 51 female) aged 8-16.9 yrs underwent an identical exercise protocol. Following studies at rest, they initially bicycled at 25 W x m(-2), increasing every 3 min by 15 W x m(-2) until exhaustion, after which measurements were made during recovery. Effective pulmonary blood flow, stroke volume, oxygen consumption, arteriovenous oxygen difference and functional residual capacity (FRC) together with estimates of pulmonary capillary blood volume and transit time were assessed at every exercise stage using inert gas rebreathing techniques. Haemodynamic performance is highly dependent on surface area, age, gender and pubertal stage. Many parameters, for example transfer factor, demonstrate pubertal stage-dependent differences at identical workloads even after correction for size. Females have a lower capillary blood volume at rest compared to age and size-matched males, but it is equalized during exercise. FRC unexpectedly rose with exercise, and peak exercise was associated with a falling stroke volume in 91% (95% CI 84-96%) of children, a possible demonstration of Starling's law of the heart. Oxygen pulse (oxygen consumption/cardiac frequency) is a very poor marker for pulmonary blood flow. Normal values are provided for all haemodynamic parameters for rest and every exercise stage for all subgroups of children. This should allow accurate comparison of normal and disease groups in future.


Perceptions of upper-body problems during recovery from breast cancer treatment.
Despite improved recognition recently, restrictions in upper-body movement continue to cause impairment and distress for many women long after breast cancer treatment. The purpose of this research is to investigate this issue through the perceptions of breast cancer survivors in the context of their everyday lives. Twenty-four women recruited from a private breast clinic in south-eastern Queensland, Australia, participated in a qualitative study. Discussion groups comprised women treated for breast cancer within the previous 18 months. Discussions centred on experiences of physical difficulties, follow-up support, arm lymphoedema and exercise therapy during the women's recoveries. Returning to normal activities for women after breast surgery was felt to take longer than either the women's or their physicians' expectations. Many women reported difficulties in upper-body tasks, which worsened simple everyday responsibilities. The physical impact leads to psychological strain, as the women are constantly reminded of their illness and the possibility they may never return to their full capacity. These upper-body difficulties may include discomfort while driving and sleeping, posture disturbances, reduced employability in physical work, and decreased ability to do housework and gardening. Having lymphoedema or the threat of developing it was very distressing for most women. The potential preventive role of physiotherapy-led exercises to prevent further decline and improve function was strongly emphasised during these discussions. Clinicians need to recognise that it is very common for women with breast cancer to experience upper-body morbidity long after their treatment, and consequently every effort to enhance recovery and avoid further deterioration in function is required.


Economic change and sex-specific cardiovascular mortality in Britain 1955-1976.
Analyses directed toward recent declines in cardiovascular disease mortality rates have typically focused on alterations in important physiological and behavioral risk factors resulting from lifestyle changes and medical advances. In this study, a multivariate model of the impact of more fundamental changes in the socioeconomic and bio-physical environments has been developed and applied to cardiovascular disease mortality rates, by sex, in England and Wales and Scotland during 1955-1976. The predictive model includes factors associated with (1) long-term growth in the economy, (2) deleterious behavioral risk factors loosely associated with economic growth--especially cigarette consumption per capita, (3) economic instability--especially recession as indicated by factors related to unemployment, income loss, and recessional declines in average weekly hours worked in manufacturing industries, (4) health care, and (5) physical environmental disturbances--especially very cold temperatures. This model proves to be an excellent instrument for the statistical explanation of trends and fluctuations in CVD mortality rates for both sexes and both regions in Britain in the post-War period. In general, the overall exponential rate of economic growth is found to be the most powerful factor in the long-term decline in CVD mortality rates. Similarly, disturbances to the national and regional economic situations--especially recessions--have regularly been associated with elevated death rates for all populations observed. Cigarette and unusually high spirits consumption, as well as particularly cold winter temperatures, have also had important deleterious effects on CVD mortality. The proportion of government expenditures devoted to health care is associated with a reduction in CVD mortality in England and Wales.


The cancer registry in the monitoring of cervical screening practice: the neglected value of the Pap smear in patients diagnosed with CIN3 on biopsy.
Patients diagnosed with CIN3 on biopsy without a Pap smear are an inevitable correlate of the extensive utilization of colposcopy as a basic screening test. In a series of 248 CIN3 cases diagnosed on biopsy and notified to the Romagna Cancer Registry between 1986 and 1993, we evaluated the independent association between the Pap smear result and (1) the final histology diagnosis, (2) the first choice treatment, and (3) the appropriateness of hysterectomy. The patients were grouped into the following categories: (1) no smear, ie colposcopy only (n = 40 or 16%), (2) CIN1, CIN2, LGSIL, and minor changes, (3) CIN3 and HGSIL, and (4) carcinoma. In multiple logistic regression analysis, patients without a Pap smear were used as a reference category. Patients with smears reported as CIN1-2/LGSIL or less as well as CIN3/HGSIL had a significantly lower probability of invasive squamous carcinoma (odds ratio = 0.26 for both categories) compared with those not undergoing cytology. The Pap smear result was not a determinant of the first choice treatment. Hysterectomy had a greater probability of being unnecessary if performed in patients with smears reported as showing CIN3/HGSIL (odds ratio = 5.05) or less. In conclusion, this registry-based study demonstrated that the Pap smear had an independent but neglected value in the assessment of cases reported as CIN3 on colposcopy-guided biopsy.


Intracellular HMGB1 as a novel tumor suppressor of pancreatic cancer.
Pancreatic ductal adenocarcinoma (PDAC) driven by oncogenic K-Ras remains among the most lethal human cancers despite recent advances in modern medicine. The pathogenesis of PDAC is partly attributable to intrinsic chromosome instability and extrinsic inflammation activation. However, the molecular link between these two events in pancreatic tumorigenesis has not yet been fully established. Here, we show that intracellular high mobility group box 1 (HMGB1) remarkably suppresses oncogenic K-Ras-driven pancreatic tumorigenesis by inhibiting chromosome instability-mediated pro-inflammatory nucleosome release. Conditional genetic ablation of either single or both alleles of HMGB1 in the pancreas renders mice extremely sensitive to oncogenic K-Ras-driven initiation of precursor lesions at birth, including pancreatic intraepithelial neoplasms, intraductal papillary mucinous neoplasms, and mucinous cystic neoplasms. Loss of HMGB1 in the pancreas is associated with oxidative DNA damage and chromosomal instability characterized by chromosome rearrangements and telomere abnormalities. These lead to inflammatory nucleosome release and propagate K-Ras-driven pancreatic tumorigenesis. Extracellular nucleosomes promote interleukin 6 (IL-6) secretion by infiltrating macrophages/neutrophils and enhance oncogenic K-Ras signaling activation in pancreatic lesions. Neutralizing antibodies to IL-6 or histone H3 or knockout of the receptor for advanced glycation end products all limit K-Ras signaling activation, prevent cancer development and metastasis/invasion, and prolong animal survival in Pdx1-Cre;K-RasG12D/+;Hmgb1-/- mice. Pharmacological inhibition of HMGB1 loss by glycyrrhizin limits oncogenic K-Ras-driven tumorigenesis in mice under inflammatory conditions. Diminished nuclear and total cellular expression of HMGB1 in PDAC patients correlates with poor overall survival, supporting intracellular HMGB1 as a novel tumor suppressor with prognostic and therapeutic relevance in PDAC.


The redox state of the [2Fe-2S] clusters in SoxR protein regulates its activity as a transcription factor.
SoxR protein is a redox-responsive transcription factor that governs a regulon of oxidative stress and antibiotic resistance genes in Escherichia coli. Purified SoxR contains oxidized [2Fe-2S] clusters and stimulates in vitro transcription of its target gene soxS up to 100-fold. SoxR transcriptional activity, but not DNA binding, is completely dependent on the [2Fe-2S] clusters; apo-SoxR prepared in vitro binds the soxS promoter with unchanged affinity but does not have transcription activity. Thus, modulation of the SoxR [2Fe-2S] clusters was proposed to control the protein's function in transcription. Here, we provide evidence that SoxR with reduced [2Fe-2S] clusters is inactive. Redox titration of purified SoxR revealed a midpoint potential of -285 +/- 10 mV (pH 7.6). In vitro transcription assays showed that SoxR was inactivated when the [2Fe-2S] cluster was reduced (-380 mV), and full activity was restored upon reoxidation (+100 mV). The results suggest that one-electron oxidation and reduction of the [2Fe-2S] cluster regulate SoxR transcriptional activity.


Biomimetics for early stage biofouling prevention: templates from insect cuticles.
A biomimetic antifouling material study was carried out utilising superhydrophobic cicada and dragonfly wings replicated with a polymer (epoxy resin). They were tested in a marine biofouling study for up to 1 week in addition to biofouling assays of protein, carbohydrate and DNA absorption. The materials were compared against a commercial antifouling paint and a polymeric smooth surface constituting a control sample. The replicated surfaces demonstrated superior antifouling properties in comparison to the control and similar efficiency in DNA (10% reduction), protein and carbohydrate adsorption (15%) to the commercial anti-fouling paint. As the fabricated surfaces have roughness at the nanometre scale it is probable that the low adsorption properties, at least in the early stages, may be related to air trapped at the surface. Interestingly the most disordered replicated surface (dragonfly wing replicate) demonstrated the lowest values of absorption.


Capitalizing strategic planning costs to recognize future value.
As healthcare organizations devote more resources to strategic planning, financial managers should consider capitalizing, rather than expensing, planning costs. Traditionally, healthcare organizations have absorbed these costs in the year a plan is developed. However, a strategic plan may be viewed as an intangible asset that provides the organization with future benefits. Therefore, its costs can be amortized over the plan's life.


Organ distribution of thorium in thorium workers: good agreement with new models of the International Commission on Radiological Protection.
Higher than environmental levels of (232)Th have been found in autopsy samples of lungs and other organs from four former employees of a thorium refinery. Working periods of the subjects ranged from 3 to 24 years, and times from end of work to death ranged from 6 to 31 years. Examination of the distribution of thorium among the organs showed that concentrations in the lung relative to pulmonary lymph nodes, bone or liver were much higher calculated from the dosimetric models in Publication 30 of the International Commission on Radiological Protection (ICRP). Much better agreement was found with more recently proposed models in Publications 68 and 69 of the ICRP. Radiation doses estimated from the amounts of thorium in the autopsy samples were compatible with health studies that found no significant difference in mortality from that of the general population of men in the U.S.


Sub1A is an ethylene-response-factor-like gene that confers submergence tolerance to rice.
Most Oryza sativa cultivars die within a week of complete submergence--a major constraint to rice production in south and southeast Asia that causes annual losses of over US 1 billion dollars and affects disproportionately the poorest farmers in the world. A few cultivars, such as the O. sativa ssp. indica cultivar FR13A, are highly tolerant and survive up to two weeks of complete submergence owing to a major quantitative trait locus designated Submergence 1 (Sub1) near the centromere of chromosome 9 (refs 3, 4, 5-6). Here we describe the identification of a cluster of three genes at the Sub1 locus, encoding putative ethylene response factors. Two of these genes, Sub1B and Sub1C, are invariably present in the Sub1 region of all rice accessions analysed. In contrast, the presence of Sub1A is variable. A survey identified two alleles within those indica varieties that possess this gene: a tolerance-specific allele named Sub1A-1 and an intolerance-specific allele named Sub1A-2. Overexpression of Sub1A-1 in a submergence-intolerant O. sativa ssp. japonica conferred enhanced tolerance to the plants, downregulation of Sub1C and upregulation of Alcohol dehydrogenase 1 (Adh1), indicating that Sub1A-1 is a primary determinant of submergence tolerance. The FR13A Sub1 locus was introgressed into a widely grown Asian rice cultivar using marker-assisted selection. The new variety maintains the high yield and other agronomic properties of the recurrent parent and is tolerant to submergence. Cultivation of this variety is expected to provide protection against damaging floods and increase crop security for farmers.


A retrospective study of risk and prognostic factors in relation to lower respiratory tract infection in elderly lung cancer patients.
Lung cancer (LCa) is one of the most common and deadly malignancies in elderly patients. During the course of the disease, these patients frequently present with lower respiratory tract infection. Therefore, this study aims to investigate the clinical features of lower respiratory tract infection in elderly LCa patients and evaluate the impact on overall survival rate. Clinical and laboratory data were analyzed retrospectively for a total of 1936 patients that were over 60-years-old. Patients were classified into three groups based on pulmonary diseases: Group 1, lung cancer (LCa); Group 2, chronic obstructive pulmonary disease (COPD); and Group 3, other medical diseases without pulmonary problems (OMD). Univariate and multivariate analysis were used to evaluate related risk factors of infections and prognostic factors. The infection rate of the LCa group (46.25%) was significantly higher than the COPD (31.40%) and OMD (23.33%) groups. Polymicrobial infections were most prevalent in the LCa group (28.75%), which far exceeded the prevalence in COPD (11.05%) and OMD (4.44%) groups. In LCa patients, the most frequent pathogens were Gram-negative bacteria (44.87%), followed by fungi (34.62%) and Gram-positive bacteria (20.51%), the major pattern of polymicrobial infections was mixed Gram-negative bacteria and fungi (43.48%). Multivariate analysis revealed that COPD, pleural effusion, anatomical type, low cellular immune function, and length of hospital stay were related risk factors of lower respiratory tract infection in elderly LCa patients. A multivariate Cox proportional hazards regression model revealed that age, stage of TNM, surgical resection, antitumor therapy, lower respiratory tract infection, COPD, and pleural effusion were independent prognostic factors for cancer-related death. Patients who received effective antimicrobial treatment had a better outcome than those who did not respond to antimicrobial drugs (HR = 0.458, P < 0.05). Understanding lower respiratory tract infection in elderly LCa patients is vital if we are to set up corresponding measures and to target effective antimicrobial treatment.


A quantitative light microscopic analysis and ultrastructural description of cholecystokinin-like immunoreactivity in the spinal trigeminal nucleus of the rat.
The spinal trigeminal nucleus is involved in orofacial sensory transmission. Cholecystokinin octapeptide has been identified in axons in this nucleus and appears to play a role in the transmission of orofacial sensation from the trigeminal ganglia to the spinal trigeminal nucleus. Although cholecystokinin has been reported in axonal processes within the spinal trigeminal nucleus at the light microscopic level, nothing is known about the synaptic relationships of these cholecystokinin axons. The goals of this study were to quantitatively determine the volume fraction of cholecystokinin-like immunoreactive cell bodies and fibers in the three subnuclei of the spinal trigeminal nucleus, to provide the first ultrastructural description of cholecystokinin-like immunoreactive processes within these subnuclei and to analyse the synaptic relationships of cholecystokinin-like immunoreactive processes within the spinal trigeminal nucleus neuropil. Cholecystokinin-like immunoreactivity was localized by the peroxidase-antiperoxidase method or the peroxidase labeled, avidin-biotin technique and quantified at the light microscopic level by point counting. Immunoreactive fibers were present in all three subnuclei, but the greatest volume fraction of immunoreactive axons was obtained in laminae I and II of the nucleus caudalis. No immunoreactive cell bodies were evident in any of the subnuclei. The majority of immunoreactive profiles in all three subnuclei were identified ultrastructurally as axon terminals that contained both small and medium sized agranular vesicles and infrequently, large dense core vesicles. These immunoreactive terminals were usually found in close contact with non-immunoreactive dendrites with which they were observed to form asymmetric synapses. Immunoreactive terminals were occasionally observed to contact the cell bodies of large non-immunoreactive neurons on the border of laminae I and II in the nucleus caudalis. These results indicate that cholecystokinin-like immunoreactive processes are present throughout the spinal trigeminal nucleus, and in nucleus caudalis show a distribution similar to that reported for the spinal cord dorsal horn. Immunoreactive axons make synaptic contact with both the dendrites and perikarya of spinal trigeminal nucleus neurons. No axoaxonic synapses were observed. These findings suggest that cholecystokinin plays an important role in spinal trigeminal nucleus function. The possible colocalization of cholecystokinin and substance P in the spinal trigeminal nucleus, and the possible role of cholecystokinin in attenuating the action of opioids in the spinal trigeminal nucleus are also discussed.


Identifiers of predominantly Spanish-speaking children with language impairment.
The purpose of this study was to identify a set of measures that would discriminate 31 predominantly Spanish-speaking children with normal language (NL children) from 31 children with language impairment (LI children). The LI children were identified as such by experienced, bilingual (Spanish/English), ASHA-certified, speech-language pathologists who were currently seeing the children in their caseloads. Children ranged in age from 5 to 7 years and were matched for age, gender, and school. Additionally, nonverbal cognitive measures assured that they did not differ significantly intellectually. Measures of vocabulary, novel bound-morpheme learning skills, and language form were randomly administered to all children. Further, parents responded to questions about their perceptions of their children's speech and language skills and family history of speech and language problems. A stepwise discriminant analysis indicated that 4 measures discriminated the groups of children with a sensitivity of 91.3% and a specificity of 100% (p < .0001): parental report of the child's speech and language skills, number of errors per T-unit, mean length per T-unit, and family history of speech and language problems. A second discriminant analysis indicated that the sensitivity and specificity could be maintained when only the first 2 measures were included. Confirmatory discriminant analyses of the 2- and 4-measure models indicated that the discriminant accuracy was stable on an independent sample.


Application of OECI accreditation program to cancer institutes of the Alleanza Contro il Cancro Italian Network: a special project supported by the Italian Ministry of Health.
The Ministry of Health of the Italian government requires for Italian Comprehensive Cancer Centers, called Istituti di Ricovero e Cura a Carattere Scientifico (IRCCS), to be certified according to international models. In this perspective, considering of particular interest the recent model for accreditation of cancer centers developed by the Organisation of European Cancer Institutes (OECI), the network of Italian IRCCS participating in Alleanza Contro il Cancro activated a project, supported in 2010 by the Ministry of Health, with the primary objective to verify the applicability of the OECI model to the Italian network of IRCCS. All 11 cancer IRCCS took part in the project. We describe in detail the activities performed, the results reached, and the impact of the initiative on the National Health Service. The initiative was concluded in November 2015 with the certification of all the Italian cancer IRCCS. Italy is the only European country that has applied the modern and tailored accreditation model of OECI to all their cancer centers.


Differential investment into testes and sperm production in alternative male reproductive tactics of the African striped mouse (Rhabdomys pumilio).
Males that follow alternative reproductive tactics might differ in their investment into testis development and sperm production. The resource-allocation hypothesis predicts that males following a sneaker tactic should invest more into sperm production than dominant territorial males which should invest more into mate guarding. This hypothesis is supported by studies in species where individual males cannot switch between tactics (fixed tactics). Here we present the first data for a species where males can switch between tactics (plastic tactics). We studied African striped mice (Rhabdomys pumilio) in captivity, mimicking three tactics observed in the field: philopatric group-living males, singly-housed males representing roaming males, and group-living breeding males. We measured quantitative and qualitative reproductive traits, as well as serum and testis hormone concentrations. We found no support for the resource-allocation hypothesis, since breeding and singly-housed males invested similarly in testes and sperm. However, philopatric males had significantly smaller testes and epididymides, lower sperm counts, lower testosterone and higher corticosterone levels than males of the two other tactics. Philopatric males did not reach a larger body mass than singly-housed males with well developed reproductive traits, indicating that they did not trade investment in sperm production against growth. Interestingly, testis testosterone concentrations of philopatric males did not differ from those of other males. Our data suggest that philopatric males are reproductively suppressed by the breeding male, but might be ready to increase their serum testosterone levels when social and environmental conditions allow for this physiological switch accompanying the behavioral switch between tactics.


Socioeconomic Status, Family Processes, and Individual Development.
Research during the past decade shows that social class or socioeconomic status (SES) is related to satisfaction and stability in romantic unions, the quality of parent-child relationships, and a range of developmental outcomes for adults and children. This review focuses on evidence regarding potential mechanisms proposed to account for these associations. Research findings reported during the past decade demonstrate support for an interactionist model of the relationship between SES and family life, which incorporates assumptions from both the social causation and social selection perspectives. The review concludes with recommendations for future research on SES, family processes and individual development in terms of important theoretical and methodological issues yet to be addressed.


A new measure for the revised reinforcement sensitivity theory: psychometric criteria and genetic validation.
Jeffrey Gray's Reinforcement Sensitivity Theory (RST) represents one of the most influential biologically-based personality theories describing individual differences in approach and avoidance tendencies. The most prominent self-report inventory to measure individual differences in approach and avoidance behavior to date is the BIS/BAS scale by Carver and White (1994). As Gray and McNaughton (2000) revised the RST after its initial formulation in the 1970/80s, and given the Carver and White measure is based on the initial conceptualization of RST, there is a growing need for self-report inventories measuring individual differences in the revised behavioral inhibition system (BIS), behavioral activation system (BAS) and the fight, flight, freezing system (FFFS). Therefore, in this paper we present a new questionnaire measuring individual differences in the revised constructs of the BIS, BAS and FFFS in N = 1814 participants (German sample). An English translated version of the new measure is also presented and tested in N = 299 English language participants. A large number of German participants (N = 1090) also filled in the BIS/BAS scales by Carver and White (1994) and the correlations between these measures are presented. Finally, this same subgroup of participants provided buccal swaps for the investigation of the arginine vasopressin receptor 1a (AVPR1a) gene. Here, a functional genetic polymorphism (rs11174811) on the AVPR1a gene was shown to be associated with individual differences in both the revised BIS and classic BIS dimensions.


3D Printed Customizable Microsampling Devices for Neuroscience Applications.
Multifunctional devices that incorporate chemical or physical measurements combined with ways to manipulate brain tissue via drug delivery, electrical stimulation, or light for optogenetics are desired by neuroscientists. The next generation in vivo brain devices will likely utilize the extensive flexibility and rapid processing of 3D printing. This Perspective demonstrates how close we are to this reality for advanced neuroscience measurements. 3D printing provides the opportunity to improve microsampling-based devices in ways that have not been previously available. Not only can 3D printing be used for actual device creation, but it can also allow printing of peripheral objects necessary to assemble functional devices. The most probable 3D printing set up for microsampling devices with appropriate nm to μm feature size will likely require 2-photon polymerization-based printers. This Perspective describes the advantages and challenges for 3D printing of microsampling devices as an initial step to meet the next generation device needs of neuroscientists.


Peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha) stimulates VLDL assembly through activation of cell death-inducing DFFA-like effector B (CideB).
Abnormalities in very low density lipoprotein (VLDL) assembly and secretion impact intrahepatic lipid homeostasis, plasma lipoprotein profile, and energy metabolism of distal peripheral tissues. We have evaluated the role of the transcriptional coactivator, the peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha), in VLDL assembly and secretion. PGC-1alpha overexpression in HepG2 cells led to diminished rates of triglyceride (TG) synthesis but strongly stimulated VLDL-TG secretion, markedly increasing the efficiency of secretion of newly synthesized TG. PGC-1alpha overexpression increased the rate of secretion of apoB100 and promoted secretion of larger, less dense VLDL particles. PGC-1alpha overexpression in intact mouse liver also stimulated rates of VLDL TG secretion and attenuated hepatic TG accumulation resulting from high fat diet feeding. To determine the molecular mechanisms mediating the effect of PGC-1alpha on VLDL assembly, we evaluated the expression of several candidate mediators known to be involved in VLDL assembly or hepatic lipid homeostasis. Cell death-inducing DFFA-like effector B (CideB) expression was greatly induced by PGC-1alpha, and siRNA against CideB reversed the effects of PGC-1alpha on the secretion of TG and VLDL-sized particles by HepG2 cells, indicating that CideB is a critical mediator of stimulatory effects of PGC-1alpha on VLDL secretion. Collectively, these data suggest that PGC-1alpha plays an important role in partitioning cytoplasmic TG toward the VLDL secretory compartments and promoting VLDL secretion via transcriptional induction of CideB.


Selenium Bioaccessibility and Speciation in Selenium-Enriched Lettuce: Investigation of the Selenocompounds Liberated after in Vitro Simulated Human Digestion Using Two-Dimensional HPLC-ICP-MS.
The evaluation of selenium-enriched vegetables as potential dietary sources of selenium, an essential element for humans, requires an assessment of the plant's accumulation ability as well as of the bioaccessibility and speciation of the accumulated selenium, which influence its biological effects in humans. Lettuce hydroponically grown at three selenite (SeVI)/selenate (SeIV) amendment levels was characterized accordingly. Selenium accumulation in lettuce leaves was greatest with Se(VI) amendment, whereas bioaccessibility was 70% on average in both cases. Selenium speciation in gastrointestinal hydrolysates, characterized by anion and cation exchange HPLC-ICP-MS, showed that Se(IV) was largely biotransformed into organoselenium metabolites, with selenomethionine accounting for 1/3 of the total detected species, whereas Se(VI) was incorporated as such in the edible portion of the plant, with only a small fraction (∼20%) converted into organic species. Taking into account both nutritional quality and safety, the Se(IV)-enriched lettuce appeared more favorable as a potential selenium source for human consumption.


Blood flow suppresses vascular Notch signalling via dll4 and is required for angiogenesis in response to hypoxic signalling.
The contribution of blood flow to angiogenesis is incompletely understood. We examined the effect of blood flow on Notch signalling in the vasculature of zebrafish embryos, and whether blood flow regulates angiogenesis in zebrafish with constitutively up-regulated hypoxic signalling. Developing zebrafish (Danio rerio) embryos survive via diffusion in the absence of circulation induced by knockdown of cardiac troponin T2 or chemical cardiac cessation. The absence of blood flow increased vascular Notch signalling in 48 h post-fertilization old embryos via up-regulation of the Notch ligand dll4. Despite this, patterning of the intersegmental vessels is not affected by absent blood flow. We therefore examined homozygous vhl mutant zebrafish that have constitutively up-regulated hypoxic signalling. These display excessive and aberrant angiogenesis from 72 h post-fertilization, with significantly increased endothelial number, vessel diameter, and length. The absence of blood flow abolished these effects, though normal vessel patterning was preserved. We show that blood flow suppresses vascular Notch signalling via down-regulation of dll4. We have also shown that blood flow is required for angiogenesis in response to hypoxic signalling but is not required for normal vessel patterning. These data indicate important differences in hypoxia-driven vs. developmental angiogenesis.


Clean indoor air laws, cigarette excise taxes, and smoking: Results from the current population survey-tobacco use supplement, 2003-2011.
There was an increase in the number and coverage of state and local clean indoor air laws in the US during the past fifteen years. These laws coincided with increases in federal, state, and local cigarette excise taxes. In light of these changes, the objective of this study was to examine the association between clean indoor air laws, cigarette excise taxes and smoking patterns between 2003 and 2011. Using data on 62,165 adult participants in the 2003 and 2010/2011 Current Population Survey-Tobacco Use Supplement who reported smoking cigarettes in the past year, we examined the association of state and county workplace, bar, and restaurant clean indoor air laws and cigarette excise taxes with quitting and current every-day smoking. Between 2003 and 2011, quitting increased and daily smoking among those who continued to smoke decreased significantly. Participants living in states and counties with higher excise taxes and more comprehensive clean indoor air laws had a higher likelihood of quitting and lower likelihood of everyday smoking. Based on the assumption of no uncontrolled confounding, changes in taxes and laws accounted for 64.8% of the increase in smoking cessation and all of the reduction in everyday smoking. Implementation of state and county-level clean indoor air laws and cigarette taxes appears to have achieved the intended goal of encouraging smokers to either quit or reduce their frequency of smoking.


Size and surface functionalization of iron oxide nanoparticles influence the composition and dynamic nature of their protein corona.
Nanoparticles (NPs) adsorb proteins when in the biological matrix, and the resulted protein corona could affect NP-cell interactions. The corona has a dynamic nature with the adsorbed proteins constantly exchanging with the free proteins in the matrix at various rates. The rapidly exchanging proteins compose the soft corona, which responds more dynamically to environment changes than the hard corona established by the ones with slow exchange rates. In the present study, the corona formed on the superparamagnetic iron oxide NPs (SPIONs) in human serum was studied by flow field-flow fractionation and ultracentrifugation, which rapidly differentiated the corona proteins based on their exchange rates. By varying the surface hydrophobicity of the SPIONs with a core size around 10 nm, we found out that, the more hydrophobic surface ligand attracted proteins with higher surface hydrophobicity and formed a more dynamic corona with a larger portion of the involved proteins with fast exchange rates. Increasing the core diameter of the SPIONs but keeping the surface ligand the same could also result in a more dynamic corona. A brief investigation of the effect on the cellular uptake of SPIONs using one selected corona protein, transferrin, was conducted. The result showed that, only the stably bound transferrin could significantly enhance cellular uptake, while transferrin bound in a dynamic nature had negligible impact. Our study has led to a better understanding of the relationship between the particle properties and the dynamic nature of the corona, which can help with design of nanomaterials with higher biocompatibility and higher efficacy in biosystems for biomedical applications.


Highlights from the 58th meeting of the American Society of Haematology, 1-6 December 2016, San Diego, USA.
The recent 58th Annual American Society of Haematology (ASH) meeting held in San Diego shed light on the usual mixture of groundbreaking basic and translational science and the recent practice-changing clinical trials. Recurrent themes this year were the use of recent next-generation sequencing (NGS) techniques to perfect prognostic stratification and disease monitoring. Newer prospects on the role of metabolism in normal and malignant haemopoiesis and mature data on long-awaited trials on immunotherapy and CAR-T cells in lymphoid neoplasms were also discussed.


Ancestral hemoglobins in Archaea.
Hemoglobins are ubiquitous in Eukarya and Bacteria but, until now, have not been found in Archaea. A phylogenetic analysis of the recently revealed microbial family of globin-coupled heme-based sensors suggests that these sensors descended from an ancient globin-only progenitor, or a protoglobin (Pgb). Here, we report the discovery and characterization of two Pgbs from the Archaea: ApPgb from the obligately aerobic hyperthermophile Aeropyrum pernix, and MaPgb from the strictly anaerobic methanogen Methanosarcina acetivorans. Both ApPgb and MaPgb bind molecular oxygen, nitric oxide, and carbon monoxide by means of a heme moiety that is coordinated to the protein through the F8 histidine (histidine 120). We postulate that these archaeal globins are the ancestors of contemporary hemoglobins.


The international anorectal physiology working group (IAPWG) recommendations: Standardized testing protocol and the London classification for disorders of anorectal function.
This manuscript summarizes consensus reached by the International Anorectal Physiology Working Group (IAPWG) for the performance, terminology used, and interpretation of anorectal function testing including anorectal manometry (focused on high-resolution manometry), the rectal sensory test, and the balloon expulsion test. Based on these measurements, a classification system for disorders of anorectal function is proposed. Twenty-nine working group members (clinicians/academics in the field of gastroenterology, coloproctology, and gastrointestinal physiology) were invited to six face-to-face and three remote meetings to derive consensus between 2014 and 2018. The IAPWG protocol for the performance of anorectal function testing recommends a standardized sequence of maneuvers to test rectoanal reflexes, anal tone and contractility, rectoanal coordination, and rectal sensation. Major findings not seen in healthy controls defined by the classification are as follows: rectoanal areflexia, anal hypotension and hypocontractility, rectal hyposensitivity, and hypersensitivity. Minor and inconclusive findings that can be present in health and require additional information prior to diagnosis include anal hypertension and dyssynergia. This framework introduces the IAPWG protocol and the London classification for disorders of anorectal function based on objective physiological measurement. The use of a common language to describe results of diagnostic tests, standard operating procedures, and a consensus classification system is designed to bring much-needed standardization to these techniques.


Inhibition of endotoxin-induced macrophage chemokine production by VIP and PACAP in vitro and in vivo.
Inflammatory chemokines recruit immune cells which initiate and maintain the inflammatory response. Although such a response is necessary for the elimination of the antigen, the inflammation has to be eventually resolved. Peptides such as vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP), released following antigenic stimulation, contribute to the termination of an inflammatory response primarily by inhibiting the production of proinflammatory cytokines. Here we investigated the effects of VIP and PACAP on chemokine production. We report that VIP and PACAP inhibit the expression of the macrophage-derived CXC chemokines MIP-2 and KC (IL-8), and of the CC chemokines MIP-1a, MIP-1b, MCP-1 and RANTES in vivo and in vitro. The decrease of chemokine gene expression correlates with an inhibitory effect of VIP/PACAP on NFkB binding. In an in vivo model of acute peritonitis, the inhibition of chemokine production by VIP/PACAP leads to a significant reduction in the recruitment of PMNs, macrophages and lymphocytes into the peritoneal cavity. These findings support the proposed role of VIP and PACAP as key endogenous anti-inflammatory agents, and describe a novel mechanism, i.e., the inhibition of the production of macrophage-derived chemokines.


Emergent surgical treatment of radiation-induced enteropathies for patients with urogynecological and colorectal carcinomas.
The aim of this study is to describe our 10-year experience in patients with urogynecological and colorectal carcinomas with radiation enteropathy treated surgically as an emergency, and to reassess symptoms and mortality. The study included 17 patients receiving emergency surgery for complications of radiotherapy. Data about the patients and outcomes of the treatment alternatives used were retrospectively analyzed. Of 17 patients, nine had colorectal cancer, six had gynecological cancer, and two had cancer of the urinary system. As an emergency, 12 patients had ileus only, one patient had intestinal fistulae and ileus, two patients had bleeding and ileus and two patients had perforation on admission. Seven patients underwent resection and anastomosis, two patients intestinal by-pass, four patients resection and ostomy and four patients bridectomy. Morbidity (75% for early complications and 100% for late complications) and mortality in the early postoperative period (25%) were higher in the patients undergoing bridectomy than in the patients undergoing other surgical methods. The rate of early and late complications (71.4% and 66.6%, respectively) was lower in the patients undergoing resection-anastomosis with a higher of quality life. Only 11 patients survived during a long follow-up period (64.7%). As the postoperative complication rate, overall and operative mortality of patients treated for radiation enteropaties as emergent surgery are high, specialists following this group of patients may favor removal of the pathologic tissue to avoid complications in the early and late postoperative period.


[Ocular manifestations of cancer].
Cancer may affect the eye and orbit as a direct result of metastatic neoplastic infiltration, compression, or circulating antibodies involving paraneoplastic retinal degeneration. A metastatic tumor to the uvea is the most common form of an intraocular metastatic process. The choroid is the most common site for uveal metastasis; metastases to the ciliary body, iris, retina, optic disk, and vitreous are rare. Approximately one-third of patients have no history of primary cancer at the time of ocular diagnosis. Breast and lung carcinomas for women and lung and gastrointestinal carcinomas for men most commonly metastasize to the eye and orbit. The short-term prognosis for vision is usually good after an individualized therapeutic approach (chemotherapy, hormonal therapy, external beam radiotherapy, or plaque radiotherapy), but the systemic prognosis is poor. The visual paraneoplastic syndromes encompass several distinct clinical and pathological entities including carcinoma-associated retinopathy (CAR), melanoma-associated retinopathy (MAR), and bilateral diffuse melanocytic uveal proliferation (BDUMP). The CAR syndrome affects photoreceptors, MAR is thought to affect bipolar cell function, and BDUMP targets the uveal tract. Identification of circulating antibodies against retinal proteins (recovering, 23-kDa retinal protein; 46-kDa and 60-kDa retinal proteins) serves to recognize the paraneoplastic nature of the patient's symptoms, which frequently develop before the cancer is diagnosed. Anecdotal therapeutic responses are described after systemic steroids, immunoglobulin injection, and plasmapheresis. Recognition of their visual symptoms and ocular findings should alert the ophthalmologist to the possibility of cancer and systemic evaluation should be pursued.


De novo mutation rates at the single-mutation resolution in a human HBB gene region associated with adaptation and genetic disease.
Although it is known that the mutation rate varies across the genome, previous estimates were based on averaging across various numbers of positions. Here, we describe a method to measure the origination rates of target mutations at target base positions and apply it to a 6-bp region in the human hemoglobin subunit beta (HBB) gene and to the identical, paralogous hemoglobin subunit delta (HBD) region in sperm cells from both African and European donors. The HBB region of interest (ROI) includes the site of the hemoglobin S (HbS) mutation, which protects against malaria, is common in Africa, and has served as a classic example of adaptation by random mutation and natural selection. We found a significant correspondence between de novo mutation rates and past observations of alleles in carriers, showing that mutation rates vary substantially in a mutation-specific manner that contributes to the site frequency spectrum. We also found that the overall point mutation rate is significantly higher in Africans than in Europeans in the HBB region studied. Finally, the rate of the 20A→T mutation, called the "HbS mutation" when it appears in HBB, is significantly higher than expected from the genome-wide average for this mutation type. Nine instances were observed in the African HBB ROI, where it is of adaptive significance, representing at least three independent originations; no instances were observed elsewhere. Further studies will be needed to examine mutation rates at the single-mutation resolution across these and other loci and organisms and to uncover the molecular mechanisms responsible.


Impact of three annual tobacco tax rises on tobacco sales in remote Australian Aboriginal community stores.
There is strong evidence from many settings that tobacco tax rises which increase prices reduce tobacco consumption, but only limited evidence from Indigenous settings. We analysed 3 years (2016-2018) of weekly sales data from 32 stores in remote Aboriginal communities. We used interrupted time series analysis to estimate the immediate impact of the price rice following annual 12.5% tobacco tax rises on sales on (A) stick equivalents of tobacco and (B) fruit and vegetables (kg) per $A1000 of grocery sales, and on the trend in sales between price rises. We detected 5.8% and 8.2% immediate declines in tobacco sales following the price rises associated with annual 12.5% tax rises in 2016 and 2018, and a non-significant decline (1.6%) following the 2017 tax rise. Decreased sales were mainly driven by declines in mainstream and premium factory-made cigarettes. Fruit and vegetable sales did not change at the time of tobacco price rises. For the first time, we demonstrated evidence of price-sensitivity and the immediate impact of price rises from tobacco tax rises on tobacco sales in remote Aboriginal communities. We acknowledge that Australia already has very high tobacco taxation and prices, but recommend further increases to the taxation of roll-your-own (RYO) tobacco to prevent smokers and industry using cheaper RYO cigarettes to undermine this impact of high tobacco taxes and prices.


Subcellular localization and molecular topology of the dopamine transporter in the striatum and substantia nigra.
Plasma membrane transporters remove neurotransmitters from the extracellular space and have been postulated to terminate synaptic activity. Their specific roles in synaptic and nonsynaptic neurotransmission at a cellular level, however, remain unclear. We have determined the subcellular location of the dopamine transporter (DAT) by immunoperoxidase and immunogold electron microscopy, using monoclonal antibodies to both the N-terminus and the second extracellular loop. The two DAT epitopes were found on opposite faces of cellular and intracellular membranes, providing confirmation of the predicted molecular topology of DAT. In the striatum, DAT was localized in the plasma membrane of axons and terminals. Double immunocytochemistry demonstrated DAT colocalization with two other markers of nigrostriatal terminals, tyrosine hydroxylase and D2 dopamine receptors. The latter was thus demonstrated to be an autoreceptor. Labeled striatal terminals formed symmetrical synapses with spines, dendrites, and perikarya. DAT was not identified within any synaptic active zones, however, even using serial section analysis. These results suggest that striatal dopamine reuptake may occur outside of synaptic specializations once dopamine diffuses from the synaptic cleft. In the substantia nigra, DAT appears to be specifically transported into dendrites, where it can be found in smooth endoplasmic reticulum, plasma membrane, and pre- and postsynaptic active zones. These localizations suggest that DAT modulates the intracellular and extracellular dopamine levels of nigral dendrites. Within the perikarya of pars compacta neurons, DAT was localized primarily to rough and smooth endoplasmic reticulum, Golgi complex, and multivesicular bodies, identifying probable sites of synthesis, modification, transport, and degradation.


Periodic mesoporous dendrisilicas.
We report the synthesis of a new class of materials called periodic mesoporous dendrisilicas. They are prepared from dendrimers with trialkoxysilyl groups at the outmost shell. Dendrimers of various cores and numbers of shells are used. The dendrisilica synthesis is based on acid- or base-catalyzed hydrolysis of the trialkoxysilyl groups and subsequent template-directed condensation of the dendrimers into an ordered template-dendrisilica nanocomposite. The template can be removed with organic solvents, producing a periodic mesoporous dendrisilica whose pore walls consist of interconnected dendrimer building blocks.


Positive outcomes following Autologous Matrix-Induced Chondrogenesis (AMIC) in the treatment of retropatellar chondral lesions: a retrospective analysis of a patient registry.
The patellofemoral joint is a challenging environment for treating chondral defects. Among the surgical options for the treatment of chondral defects, the single-stage Autologous Matrix-Induced Chondrogenesis (AMIC) procedure uses a porcine collagen I/III membrane to enhance bone-marrow stimulation. However, longer term outcomes data are rare for this specific indication. In order to provide real-world information, an ongoing registry has been established to record patient data and outcomes when AMIC is used to treat chondral and osteochondral lesions. Patient data were retrieved from an ongoing, prospective, multisite registry of patients who had undergone AMIC treatment of chondral defects. We identified 64 patients who had undergone AMIC for patellofemoral chondral defects and for whom pre-operative and at least 1 post-operative score were available were included in this retrospective data analysis. Outcomes were assessed via the KOOS, VAS pain, and the Lysholm scores. Outcomes at the post-operative time-points were analysed using a factorial ANOVA with post-hoc testing while linear regression was used to assess associations between the change in the Lysholm score and lesion size. There was a significant improvement in Lysholm, VAS pain, and KOOS scores from pre-operative to the 1st year post-operative (p < 0.001), and this was maintained during the follow-up. The forces exerted on the patellofemoral joint make this a challenging scenario for chondral repair. Our data demonstrates that the AMIC procedure with a collagen I/III membrane is an effective treatment for retropatellar cartilage lesions, and provides reliable results, with decreased pain and improved function. Importantly, these improvements were maintained through the follow-up period.


Somatic variants for seed and fruit set in grapevine.
Grapevine reproductive development has direct implications on yield. It also impacts on berry and wine quality by affecting traits like seedlessness, berry and bunch size, cluster compactness and berry skin to pulp ratio. Seasonal fluctuations in yield, fruit composition and wine attributes, which are largely driven by climatic factors, are major challenges for worldwide table grape and wine industry. Accordingly, a better understanding of reproductive processes such as gamete development, fertilization, seed and fruit set is of paramount relevance for managing yield and quality. With the aim of providing new insights into this field, we searched for clones with contrasting seed content in two germplasm collections. We identified eight variant pairs that seemingly differ only in seed-related characteristics while showing identical genotype when tested with the GrapeReSeq_Illumina_20K_SNP_chip and several microsatellites. We performed multi-year observations on seed and fruit set deriving from different pollination treatments, with special emphasis on the pair composed by Sangiovese and its seedless variant locally named Corinto Nero. The pollen of Corinto Nero failed to germinate in vitro and gave poor berry set when used to pollinate other varieties. Most berries from both open- and cross-pollinated Corinto Nero inflorescences did not contain seeds. The genetic analysis of seedlings derived from occasional Corinto Nero normal seeds revealed that the few Corinto Nero functional gametes are mostly unreduced. Moreover, three genotypes, including Sangiovese and Corinto Nero, were unexpectedly found to develop fruits without pollen contribution and occasionally showed normal-like seeds. Five missense single nucleotide polymorphisms were identified between Corinto Nero and Sangiovese from transcriptomic data. Our observations allowed us to attribute a seedlessness type to some variants for which it was not documented in the literature. Interestingly, the VvAGL11 mutation responsible for Sultanina stenospermocarpy was also discovered in a seedless mutant of Gouais Blanc. We suggest that Corinto Nero parthenocarpy is driven by pollen and/or embryo sac defects, and both events likely arise from meiotic anomalies. The single nucleotide polymorphisms identified between Sangiovese and Corinto Nero are suitable for testing as traceability markers for propagated material and as functional candidates for the seedless phenotype.


Dibenzo crown ether layer formation on muscovite mica.
Stable layers of crown ethers were grown on muscovite mica using the potassium-crown ether interaction. The multilayers were grown from solution and from the vapor phase and were analyzed with atomic force microscopy (AFM), matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, and surface X-ray diffraction (SXRD). The results show that the first molecular layer of the three investigated dibenzo crown ethers is more rigid than the second because of the strong interaction of the first molecular layer with the potassium ions on the surface of muscovite mica. SXRD measurements revealed that for all of the investigated dibenzo crown ethers the first molecule lies relatively flat whereas the second lies more upright. The SXRD measurements further revealed that the molecules of the first layer of dibenzo-15-crown-5 are on top of a potassium atom, showing that the binding mechanism of this layer is indeed of the coordination complex form. The AFM and SXRD data are in good agreement, and the combination of these techniques is therefore a powerful way to determine the molecular orientation at surfaces.


Intermetallic germanides with non-centrosymmetric structures derived from the Yb3Rh4Sn13 type.
New germanides with composition RE3Pt4Ge13 (RE = Y, Pr, Sm, Gd, Tb, Tm) have been prepared by high-pressure, high-temperature synthesis. Their crystal structures have been refined, and the relationship of this new rhombohedral and monoclinic structure types with the primitive cubic Yb3Rh4Sn13 prototype is discussed. Band structure calculations within density functional theory confirm the distorted rhombohedral and monoclinic structural arrangements to be energetically more favorable than the simple cubic one. X-ray absorption spectroscopy and magnetic susceptibility measurements indicate that the RE-atoms are in the +3 oxidation state in all studied compounds.


Effect of growth conditions on the formation of the relaxation complex of supercoiled ColE1 deoxyribonucleic acid and protein in Escherichia coli.
Colicinogenic factor E1 (ColE1) is present in Escherichia coli strain JC411 (ColE1) cells to the extent of about 24 copies per cell. This number does not appear to vary in situations which give rise to twofold differences in the amount of chromosomal deoxyribonucleic acid (DNA) present per cell. If cells are grown in the absence of glucose, approximately 80% of the ColE1 molecules can be isolated as strand-specific DNA-protein relaxation complexes. When glucose is present in the medium, only about 30% of the plasmid molecules can be isolated as relaxation complexes. Medium shift experiments in which glucose was removed from the medium indicate that within 15 min after the shift the majority (>60%) of the plasmid can be isolated as relaxation complex. This rapid shift to the complexed state is accompanied by a two- to threefold increase in the rate of plasmid replication. The burst of replication and the shift to the complexed state are both inhibited by the presence of chloramphenicol. Inhibition of protein synthesis in log cultures by the addition of chloramphenicol or amino acid starvation allows ColE1 DNA to continue replicating long after chromosomal replication has ceased. Under these conditions, noncomplexed plasmid DNA accumulates while the amount of DNA that can be isolated in the complexed state remains constant at the level that existed prior to treatment. In the presence of chloramphenicol, there appears to be a random dissociation and association of ColE1 DNA and "relaxation protein" during or between rounds of replication.


Effect of monensin withdrawal on intake, digestion, and ruminal fermentation parameters by Bos taurus indicus and Bos taurus taurus steers consuming bermudagrass hay.
Effects of monensin withdrawal and cattle subspecies on the utilization of bermudagrass hay (14.3% CP, 72.3% NDF, and 36.9% ADF) were evaluated using ruminally cannulated steers (5 [BI] and 5 [BT]). Subspecies were concurrently subjected to a 2-period, 2-treatment crossover design. Treatments consisted of either 0 mg·steer·d-1 monensin with no previous monensin feeding (CON) or withdrawal from 200 mg·steer·d-1 monensin (MON) fed individually in 0.91 kg dried distillers' grains with solubles for 42 d. Withdrawal was evaluated for a 28-d period. Ruminal fluid was collected 2 h after feeding on d 0, 1, 4, 7, 14, and 21 after withdrawal for determination of pH, VFA, ruminal NH-N (RAN), rate of NH production, and CH production rate. Hay, ort, and fecal grab samples were collected d 23 through 28 after withdrawal for determination of intake and digestion. No subspecies × monensin, subspecies × day, or subspecies × monensin × day interactions were observed ( ≥ 0.11). An effect of day after monensin withdrawal was observed ( < 0.01) for total VFA concentration, with an increase following withdrawal followed by a decrease and then stabilization. Monensin × day after monensin withdrawal interactions ( ≤ 0.01) were observed for the acetate:propionate (A:P) ratio and molar percent of acetate and propionate. There was a decrease in molar percent of propionate between d 1 and 4 from 19.1 to 18.0; however, it remained greater ( ≤ 0.10) for MON than CON through d 7. Withdrawal increased molar percent of acetate from 68.3 to 69.8 between d 0 and 4 for MON steers. The A:P ratio was less ( ≤ 0.01) on d 0 for MON than for CON (3.4 vs. 4.0), but by d 4, it increased to 3.8 and was not different ( = 0.14) from CON. By d 14, no differences ( ≥ 0.88) remained for acetate, propionate, or the A:P ratio. After monensin withdrawal, monensin reduced ( < 0.01) RAN by 12.3% (2.09 vs. 1.83 m for CON and MON, respectively). Monensin withdrawal and cattle subspecies had no effect ( ≥ 0.23) on rate of NH production or CH production rate. Monensin withdrawal had no effect ( ≥ 0.45) on intake or digestibility parameters. Greater forage OM intake ( = 0.09; 21.2 vs. 19.2 g/kg BW) and OM digestibility ( < 0.01; 72.4 vs. 63.0%) resulted in greater ( < 0.01) total digestible OM intake (16.8 vs. 13.2 g/kg BW) in BT steers than in BI steers. These results suggest that BT steers are better able to utilize bermudagrass hay than BI steers. Upon monensin withdrawal, steers previously fed monensin continue to have a reduced A:P ratio for at least 7 d.


Proapoptotic protein Bim regulates the suppressive function of Treg cells.
Regulatory T (Treg) cells are a special immunosuppressive subset of cluster of differentiation 4-positive (CD4+)-T lymphocytes and play a pivotal role in the establishment of immune homeostasis in vivo (Zhang et al., 2021). The transcription factor forkhead box protein P3 (Foxp3) is the master marker of Treg cells, which is highly expressed in Treg cells and is also essential for their suppressive function (Hori et al., 2003). In addition to Foxp3, other regulators of Treg cells have been discovered (Wu et al., 2017, 2022; Wu and Sun, 2023a, 2023b); however, a deeper understanding of the regulation of these cells is required.


[Treatment of chronic osteoarthritis pain: effectivity and safety of a 7 day matrix patch with a low dose buprenorphine].
Patients with osteoarthritis often suffer from chronic pain. If pain treatment with NSAIDS and coxibes is no longer indicated, a constant and user friendly opioid analgesia can be achieved with a low dose buprenorphine patch being applicated using an interval of 7 days. The use of this matrix patch was evaluated in a multicenter observational study on 4263 patients in clinical practice. During treatment a significant decrease of mean pain intensity on a 11-point scale could be observed from 6.9 points before using the patch to 2.9 points at the end of observation. Further effects were a decrease of additional analgetic medication and an improvement of aspects of life quality, e.g. mobility and quality of sleep. Only in 4.5% of the patients adverse effects were observed, reflecting the expected range of adverse effects of opioids. Thus it could be demonstrated that the use of the transdermal patch is an effective, user friendly and safe way of chronic pain relief for osteoarthritis patients.


Transcriptome Analysis Identifies Multifaceted Regulatory Mechanisms Dictating a Genetic Switch from Neuronal Network Establishment to Maintenance During Postnatal Prefrontal Cortex Development.
The prefrontal cortex (PFC) is one of the latest brain regions to mature, which allows the acquisition of complex cognitive abilities through experience. To unravel the underlying gene expression changes during postnatal development, we performed RNA-sequencing (RNA-seq) in the rat medial PFC (mPFC) at five developmental time points from infancy to adulthood, and analyzed the differential expression of protein-coding genes, long intergenic noncoding RNAs (lincRNAs), and alternative exons. We showed that most expression changes occur in infancy, and that the number of differentially expressed genes reduces toward adulthood. We observed 137 differentially expressed lincRNAs and 796 genes showing alternative exon usage during postnatal development. Importantly, we detected a genetic switch from neuronal network establishment in infancy to maintenance of neural networks in adulthood based on gene expression dynamics, involving changes in protein-coding and lincRNA gene expression as well as alternative exon usage. Our gene expression datasets provide insights into the multifaceted transcriptional regulation of the developing PFC. They can be used to study the basic developmental processes of the mPFC and to understand the mechanisms of neurodevelopmental and neuropsychiatric disorders. Our study provides an important contribution to the ongoing efforts to complete the "brain map", and to the understanding of PFC development.


Regulatory status of caffeine in the United States.
This article summarizes the history of the regulation of caffeine, a key component of caffeine-containing energy drinks and other caffeine-containing energy products, in the United States. Caffeine as an ingredient in food has been regulated by the US Food and Drug Administration (FDA) since 1958, when the Food Additives Amendment to the Federal Food, Drug and Cosmetic Act was enacted. It is listed as a substance that is generally recognized as safe by experts for its intended use in cola-type beverages at levels not to exceed 200 parts per million. Here, the history of FDA evaluations of the safe use of, as well as consumer exposure to, caffeine in food in the United States is outlined. Finally, the FDA's current concerns about caffeine and caffeine-containing energy products are reported, along with the current activities to address those concerns.


Examining an Australian physical activity and nutrition intervention using RE-AIM.
Translating evidence-based interventions into community practice is vital to health promotion. This study used the RE-AIM framework to evaluate the larger dissemination of the ManUp intervention, an intervention which utilized interactive web-based technologies to improve the physical activity and nutrition behaviors of residents in Central Queensland, Australia. Data were collected for each RE-AIM measure (Reach, Effectiveness, Adoption, Implementation, Maintenance) using (i) computer-assisted telephone interview survey (N = 312) with adults (18 years and over) from Central Queensland, (ii) interviews with key stakeholders from local organizations (n = 12) and (iii) examination of project-related statistics and findings. In terms of Reach, 47% of participants were aware of the intervention; Effectiveness, there were no significant differences between physical activity and healthy nutrition levels in those aware and unaware; Adoption, 73 participants registered for the intervention and 25% of organizations adopted some part of the intervention; Implementation, 26% of participants initially logged onto the website, 29 and 17% started the web-based physical activity and nutrition challenges, 33% of organizations implemented the intervention, 42% considered implementation and 25% reported difficulties; Maintenance, an average of 0.57 logins and 1.35 entries per week during the 12 week dissemination and 0.27 logins and 0.63 entries per week during the 9-month follow-up were achieved, 22 and 0% of participants completed the web-based physical activity and nutrition challenges and 33.3% of organizations intended to continue utilizing components of the intervention. While this intervention demonstrated good reach, effectiveness, adoption and implementation warrant further investigation.


COVID-19 handling report for pre-case, case (pre-hospital and hospital), and post-case phases in the elderly as vulnerable populations in 6 Asia Pacific countries.
In this current COVID-19 pandemic, the elderly (60 years and over) are more vulnerable populations to be infected and become victims. In a disaster cycle, the various parts are usually divided into three stages, consisting of the pre-impact stage, the trans-impact stage, and the post-impact stage. It is necessary to explain how to handle the COVID-19 disaster for the elderly at each step (explain the meaning of pre-case, case (pre-hospital and hospital), and post-case phases, respectively). This paper presents the handling of COVID-19 for elderly in pre-case, case, and post-case phases in six Asia-Pacific countries (Indonesia, Thailand, Singapore, Malaysia, Vietnam, and Japan). The data and information come from COVID-19 official websites of each country, including information from World Health Organization (WHO), United States Centers for Disease Control and Prevention (CDC), mass media, and professional associations. The handling of COVID-19 in the pre-case phase has been done correctly for the elderly, especially in Indonesia, Japan, Thailand, and Singapore. In the case phase (pre-hospital and hospital), only Indonesia, Japan, and Thailand have followed special handling protocols for the elderly, particularly for those who have comorbidities and respiratory diseases. For the post-case phase, all countries have the same treatment protocol for all age groups, with none specific for the elderly.


Clinical trials of pneumonia management assess heterogeneous outcomes and measurement instruments.
To inform clinical practice guidelines, randomized controlled trials (RCTs) of the management of pneumonia need to address the outcomes that are most important to patients and health professionals using consistent instruments, to enable results to be compared, contrasted, and combined as appropriate. This systematic review describes the outcomes reported in clinical trials of pneumonia management and the instruments used to measure these outcomes. Based on a prospective protocol, we searched MEDLINE/PubMed, Cochrane CENTRAL and clinical trial registries for ongoing or completed clinical trials evaluating pneumonia management in adults in any clinical setting. We grouped reported outcomes thematically and classified them following the COMET Initiative's taxonomy. We describe instruments used for assessing each outcome. We found 280 eligible RCTs of which 115 (41.1%) enrolled critically ill patients and 165 (58.9%) predominantly noncritically ill patients. We identified 43 distinct outcomes and 108 measurement instruments, excluding nonvalidated scores and questionnaires. Almost all trials reported clinical/physiological outcomes (97.5%). Safety (63.2%), mortality (56.4%), resource use (48.6%) and life impact (11.8%) outcomes were less frequently addressed. The most frequently reported outcomes were treatment success (60.7%), mortality (56.4%) and adverse events (41.1%). There was significant variation in the selection of measurement instruments, with approximately two-thirds used in less than 10 of the 280 RCTs. None of the patient-reported outcomes were used in 10 or more RCTs. This review reveals significant variation in outcomes and measurement instruments reported in clinical trials of pneumonia management. Outcomes that are important to patients and health professionals are often omitted. Our findings support the need for a rigorous core outcome set, such as that being developed by the European Respiratory Society.


Evaluation of carotid artery outward remodeling by T1-weighted magnetic resonance imaging in carotid endarterectomy and stenting.
We evaluated carotid artery outward remodeling and plaque relative signal intensity (rSI) using T1-weighted magnetic resonance imaging (T1-MRI) to investigate their clinical significance in carotid revascularization. From 86 patients undergoing carotid endarterectomy (CEA) or carotid artery stenting (CAS), 88 lesions (51 lesions treated with CEA and 37 lesions treated with CAS) were analyzed retrospectively. We evaluated the preoperative carotid artery remodeling index (CRI), determined by a ratio of the external cross-sectional vessel area at maximum stenosis and the reference cross-sectional vessel area at the distal portion of the internal carotid artery, and the plaque rSI, which is quantified as the ratio between the signal intensities of plaque and adjacent muscle using T1-MRI. We divided carotid lesions into four groups using the median values of CRI and rSI: L/L (CRI < 1.8, rSI < 2.5), H/L (CRI ≥ 1.8, rSI < 2.5), L/H (CRI < 1.8, rSI ≥ 2.5), and H/H (CRI ≥ 1.8, rSI ≥ 2.5). The primary end point was detection of acute ipsilateral ischemia on diffusion-weighted imaging (DWI) within 72 hours of treatment. Mean CRI and rSI were significantly higher in lesions treated with CEA than in those treated with CAS. Postoperative DWI abnormalities were observed in 4 CEA cases (7.8%) and 10 CAS cases (27.0%) (P = .01). In the CAS group, the frequency of DWI abnormalities was 5.5% for the L/L, 40.0% for the H/L and L/H, and 55.5% for the H/H group (P = .009). Multivariate analysis showed that the degree of stenosis and H/H lesion were independent risk factors for cerebral embolism. No correlation was found between plaque parameters and postoperative DWI findings in the CEA group. CRI and rSI provide complementary information for the prediction of high-risk plaques associated with CAS but not with CEA. Preoperative evaluation with T1-MRI facilitates the selection of a treatment strategy for carotid artery stenosis.


Impact of targeting Kt instead of Kt/V.
Patients must receive an adequate dialysis dose in each hemodialysis (HD) session. Ionic dialysance (ID) enables the dialysis dose to be monitored in each session. The aim of this study was to compare the achievement of Kt versus eKt/V values and to analyse the main impediments to reaching the dialysis dose. Of 5316 patients from 54 Fresenius Medical Care centers in Spain undergoing their usual HD regime, 3275 received ID and were included in the study. The minimum prescribed dose of eKt/V was reached in 91.2% of the patients, while the minimum recommended dose of Kt was reached in only 66.8%. Patients not receiving the minimum Kt dose were older, had spent 7 months less on dialysis, had a dialysis duration of 6 min less, had 5.7 kg more of body weight and Qb was 47 mL/min lower. The target Kt was not reached by 62% of patients with catheters and by 37% of women. With each quintile increase of body weight, eKt/V decreased and Kt increased. Of patients with a body weight >80 kg, 1.4%, mostly men, reached the target Kt but not prescribed eKt/V. The impact of monitoring the dose with Kt instead of Kt/V is that identifies 25.8% of patients who did not reach the minimum Kt while achieving Kt/V. The main impediments to achieving an adequate dialysis dose were catheter use, female sex, advanced age, greater body weight, shorter dialysis time and lower Qb.


[Metabolic activity of neutrophilic granulocytes and possible ways of its correction in patients with acute coronary syndrome].
The present study aimed to investigate the metabolic activity of neutrocytes and the action of corvitin on the level of superoxide anion and myeloperoxidases of cells in vitro with the calculation of index of consumption of myeloperoxidase in patients with ST-elevation acute coronary syndrome. Patient were divided into 2 groups according to the level of superoxide anion. Group 1 included the patients (68%) with the initially low level of superoxide anion, and adding of corvitin to the cells of such patients promoted normalization of this index. In this group we observed also neutrocytosis, low index of consumption of myeloperoxidase and a high level of this enzyme in general population of neutrocytes. Group 2 included patients (32%) with initially normal level of superoxide anion. In this group, corvitin did not influence substantially this factor. Such patients had a level ofmyeloperoxidase within control values and the index of consumption of this enzyme was also within control values. The analysis of hospital period showed that the patients of group 1 had a higher frequency of ventricular tachycardia/ventricular fibrillation, paroxysms of atrial fibrillation, bundle-branch blocks and worsening of the kidney function. We suppose that a low level of superoxide anion in neutrocytes play a major role in the development of complications in patients with acute coronary syndrome. An intravenous administration of corvitin was effective in restoring the metabolic activity of neutrocytes.


Genetic predisposition and parameters of malignant progression in K14-HPV16 transgenic mice.
Reproducible multi-stage progression to invasive squamous carcinoma of the epidermis has been achieved in transgenic mice expressing the HPV16 early-region genes, including the E6/E7 oncogenes, under the control of the human keratin-14 promoter/enhancer. Although 100% of K14-HPV16 transgenic animals develop hyperplastic and/or dysplastic lesions in several inbred backgrounds, including C57BL/6, BALB/c, and SSIN/SENCAR, only mice backcrossed into the FVB/n background progress to malignant squamous cell carcinomas of two pathological grades, well differentiated and moderate/poorly differentiated (WDSC or MPDSC, respectively), each displaying characteristic patterns of malignant behavior. WDSCs typically arise within the epidermis of the ear and invade deeply into the underlying dermis but fail to metastasize, whereas MPDSCs develop on the chest and truncal skin and invariably metastasize to regional lymph nodes. The transition to the malignant state, in 21% of FVB/n transgenic mice, is characterized by alteration of the repertoire of keratin intermediate filament proteins expressed within neoplastic epidermis, such that WDSCs maintain expression of keratins common to terminally differentiating stratified keratinocytes (K10), whereas MPDSCs are distinguished from WDSCs by activation of embryonic and mucosal keratins (K13, K8, and K19). Precursor hyperplastic and dysplastic lesions are characterized by a progressively increased proliferative index, striking morphological alterations in keratinocyte cell-cell and cell-matrix interactions, and extensive remodeling of the underlying dermal stroma. Remarkably, this extensive stromal remodeling, which may facilitate both angiogenesis and eventual tumor cell invasion, develops early at the dysplastic stage in all animals well before malignant conversion.


Dps protects cells against multiple stresses during stationary phase.
Dps, the nonspecific DNA-binding protein from starved cells, is the most abundant protein in stationary-phase Escherichia coli. Dps homologs are found throughout the bacteria and in at least one archaeal species. Dps has been shown to protect cells from oxidative stress during exponential-phase growth. During stationary phase, Dps organizes the chromosome into a highly ordered, stable nucleoprotein complex called the biocrystal. We show here that Dps is required for long-term stationary-phase viability under competitive conditions and that dps mutants have altered lag phases compared to wild-type cells. We also show that during stationary phase Dps protects the cell not only from oxidative stress but also from UV and gamma irradiation, iron and copper toxicity, thermal stress, and acid and base shock. The protective roles of Dps are most likely achieved through a combination of functions associated with the protein-DNA binding and chromosome compaction, metal chelation, ferroxidase activity, and regulation of gene expression.


Image analysis of neurofilament immunoreactivity in human spiral ganglion.
Computer-assisted image analysis was used to study the immunoreactivity to NF-L, NF-M and NF-H in human spiral ganglion cells. The concentrations, represented by the relative mean grey of NF-L, NF-M and NF-H, were calculated and compared. The cell-area mean-grey correlations for NF-L, NF-M and NF-H were analysed and calculated, showing that NF-M is more specific to the larger cells (type 1?) and NF-H is more specific for the smaller cells (type 2?), while NF-L is non-specific for cell size. These findings confirm several previous assumptions by providing a quantitative basis. We conclude that image analysis is a useful-even essential-tool for the analysis of immunostained temporal bone sections.


Pathophysiology of bone metastases: how this knowledge may lead to therapeutic intervention.
Bone metastases are common in many advanced cancers and are a clinically relevant source of skeletal morbidity. The bone mineral matrix contains numerous growth factors that are released during normal bone remodeling, providing a fertile microenvironment for tumor cell colonization and proliferation. Tumor cells then release a variety of growth factors that promote bone resorption and increase the risk of skeletal complications. Bisphosphonates are potent inhibitors of osteoclast activity that have demonstrated efficacy in the treatment of bone metastases. Bisphosphonates bind avidly to the bone matrix, are released during bone resorption, and are subsequently internalized by osteoclasts, where they interfere with biochemical pathways and induce osteoclast apoptosis. Bisphosphonates also antagonize osteoclastogenesis and promote the differentiation of osteoblasts. As a result, bisphosphonates inhibit tumor-induced osteolysis and reduce skeletal morbidity. Furthermore, preclinical studies suggest that bisphosphonates possess antitumor activity and can inhibit proliferation and induce apoptosis of tumor cell lines. In addition, zoledronic acid, a new-generation bisphosphonate, appears to inhibit tumor cell invasion of the extracellular matrix. These data suggest that zoledronic acid and other bisphosphonates may play a role in the reduction of skeletal tumor burden and the prevention of bone metastasis.


The use of povidone-iodine in ophthalmology.
Povidone-iodine (PVI) is a disinfectant and antiseptic agent used for preoperative preparation of the skin and mucous membranes, as well as for the treatment of contaminated wounds. Currently regimens for prophylaxis against postsurgical endophthalmitis are being modified, including a total withdrawal of antibiotics in intravitreal injections. The aim of this study was to evaluate the use of PVI in ophthalmology. As a result of its broad spectrum of microbicidal activity, PVI is routinely used in ophthalmic surgery. However, various protocols are applied worldwide and within different procedures. Additional indications include prophylaxis against ophthalmia neonatorum, acute conjunctivitis, adenoviral conjunctivitis, bacterial keratitis or corneal ulcer, endophthalmitis, giant fornix syndrome, and antimicrobial prophylaxis in patients with Boston type I keratoprosthesis. Despite the introduction of new antiseptics in surgery, PVI is still the preeminent antiseptic measure in ophthalmology. Its use is economically reasonable. There have been no reports of resistance to PVI or anaphylaxis with topical ophthalmic use. Furthermore, it does not induce resistance or cross-grztance to antibiotics. With these advantages the range of indications for topical use of antibiotics might decrease, with PVI as the sole perioperative antiseptic measure. Additional studies are required to assess the optimal timing, concentration and exposure time within different ophthalmic procedures.


Effect of naturally occurring triterpenoids ursolic acid and glycyrrhizic acid on the cell-mediated immune responses of metastatic tumor-bearing animals.
Effect of terpenoids, ursolic acid and glycyrrhizic acid on the cell-mediated immune response was studied in metastatic tumor-bearing C57BL/6 mice. Intraperitoneal administration of ursolic acid and glycyrrhizic acid (50 mumoles/Kg body wt for 5 consecutive days) was found to produce increased natural killer cell activity (NK-activity) in metastatic tumor-bearing animals. In the glycyrrhizic acid- and ursolic acid-treated groups the peak activity was observed on the 4(th) day (60.2% and 43% cell lysis respectively). Whereas in control animals the maximum cell lysis was obtained only on 16(th) day (25% cell lysis). Administration of terpenoids clearly enhanced the antibody dependent cell mediated cytotoxicity (ADCC). In tumor-bearing control animals maximum cell lysis was obtained only on the 16(th) day (20% cell lysis). But in the case of glycyrrhizic acid- and ursolic acid-treated groups, the maximum lysis was obtained on the 12(th) day. and it was 47% and 32.5% cell lysis, respectively. Intraperitoneal administration of these 2 terpenoids was also found to enhance antibody-dependent complement-mediated cytotoxicity (ACC) in metastatic tumor-bearing animals. The elevated level of GM-CSF in tumor-alone treated control animals (37.9 +/- 1.1 pg/ml) was reduced by the treatment with glycyrrhizic acid (20.3 pg/ml) and ursolic acid (22.5 pg/ml). The highly elevated level of IL-6 (370.1 pg/ml) in control animals was also reduced by the treatment of glycyrrhizic acid (313 pg/ml) and ursolic acid (299 pg/ml). The level of IL-2 was enhanced by the treatment with glycyrrhizic acid (37.9 pg/ml) and ursolic acid (35.9) compared to untreated tumor-bearing control animals (24.9 pg/ml).


[Electromyographic characteristics of the effectiveness of puncture reflexotherapy in vertebral osteochondrosis with neurologic syndromes].
Results of an analysis of myoelectrogenesis changes that occurred in the course of treating 240 patients with neurological syndromes of vertebral osteochondrosis are presented. The patients were given acupuncture and electro-ionic (microiontophoresis of medicinal substances into the acupuncture sites) reflexotherapy. The efficacy of these methods in the treatment of the reflectory and compression syndromes is analyzed, and so is the informative value of total electromyography in estimating the effectiveness of the treatment being given.


Variation at the TERT locus and predisposition for cancer.
Telomerase and the control of telomere length are intimately linked to the process of tumourigenesis in humans. Here I review the evidence that variation at the 5p15.33 locus, which contains the TERT gene (encoding the catalytic subunit of telomerase), might play a role in the determination of cancer risk. Mutations in the coding regions of TERT can affect telomerase activity and telomere length, and create severe clinical phenotypes, including bone marrow failure syndromes and a substantive increase in cancer frequency. Variants within the TERT gene have been associated with increased risk of haematological malignancies, including myelodysplastic syndrome and acute myeloid leukaemia as well as chronic lymphocytic leukaemia. Furthermore, there is good evidence from a number of independent genome-wide association studies to implicate variants at the 5p15.33 locus in cancer risk at several different sites: lung cancer, basal cell carcinoma and pancreatic cancer show strong associations, while bladder, prostate and cervical cancer and glioma also show risk alleles in this region. Thus, multiple independent lines of evidence have implicated variation in the TERT gene as a risk factor for cancer. The mechanistic basis of these risk variants is yet to be established; however, the basic biology suggests that telomere length control is a tantalising candidate mechanism underlying cancer risk.


Further analysis of Marek's disease virus horizontal transmission confirms that U(L)44 (gC) and U(L)13 protein kinase activity are essential, while U(S)2 is nonessential.
Marek's disease virus (MDV) causes a devastating disease in chickens characterized by the development of lymphoblastoid tumors in multiple organs and is transmitted from the skin of infected chickens. We have previously reported that the U(S)2, U(L)44 (glycoprotein C [gC]), and U(L)13 genes are essential for horizontal transmission of MDV in gain-of-function studies using an a priori spread-deficient virus that was based on an infectious clone from the highly virulent RB-1B virus (pRB-1B). To precisely determine the importance of each individual gene in the process of chicken-to-chicken transmission, we used the transmission-restored clone that readily transmits horizontally and mutated each individual gene in loss-of-function experiments. Two independent U(S)2-negative mutants transmitted horizontally, eliminating U(S)2 as being essential for the process. In contrast, the absence of gC expression or mutating the invariant lysine essential for U(L)13 kinase activity abolished horizontal spread of MDV between chickens.


DNA in ancient bone - where is it located and how should we extract it?
Despite the widespread use of bones in ancient DNA (aDNA) studies, relatively little concrete information exists in regard to how the DNA in mineralised collagen degrades, or where it survives in the material's architecture. While, at the macrostructural level, physical exclusion of microbes and other external contaminants may be an important feature, and, at the ultrastructural level, the adsorption of DNA to hydroxyapatite and/or binding of DNA to Type I collagen may stabilise the DNA, the relative contribution of each, and what other factors may be relevant, are unclear. There is considerable variation in the quality of DNA retrieved from bones and teeth. This is in part due to various environmental factors such as temperature, proximity to free water or oxygen, pH, salt content, and exposure to radiation, all of which increase the rate of DNA decay. For example, bone specimens from sites at high latitudes usually yield better quality DNA than samples from temperate regions, which in turn yield better results than samples from tropical regions. However, this is not always the case, and rates of success of DNA recovery from apparently similar sites are often strikingly different. The question arises as to whether this may be due to post-collection preservation or just an artefact of the extraction methods used in these different studies? In an attempt to resolve these questions, we examine the efficacy of DNA extraction methods, and the quality and quantity of DNA recovered from both artificially degraded, and genuinely ancient, but well preserved, bones. In doing so we offer hypotheses relevant to the DNA degradation process itself, and to where and how the DNA is actually preserved in ancient bone.


Antipsychotic Use and Stroke: A Retrospective Comparative Study in a Non-Elderly Population.
To evaluate stroke risk among users of typical antipsychotics compared to users of atypical antipsychotics in a non-elderly and non-demented US population. New users of antipsychotics aged 18-64 years without dementia were identified via electronic health care data from 13 health plans participating in the Sentinel System from January 2001 to September 2015. The risk of hospitalized stroke events, identified via ICD-9-CM diagnostic criteria, was compared between typical and atypical antipsychotic users using 1:1 matching on propensity score. Adjusted hazard ratios (HRs) and 95% CIs during the entire follow-up period and during 1- to 15-day and 16- to 90-day risk windows were estimated. The risk associated with haloperidol use was estimated separately. A total of 45,495 typical antipsychotic users were matched 1:1 to atypical antipsychotic users. While unmatched HRs suggest an increased stroke risk among typical antipsychotic users compared to atypical antipsychotic users, no increased risk was observed after matching during the entire follow-up period (HR = 0.87; 95% CI, 0.54-1.41), the 1- to 15-day risk window (HR = 1.16; 95% CI, 0.41-3.32), or the 16- to 90-day risk window (HR = 0.52; 95% CI, 0.20-1.36). The adjusted HR for haloperidol was 1.31 (95% CI, 0.54-3.21). These findings were not suggestive of an increased stroke risk in typical antipsychotic users compared to atypical antipsychotic users in a non-elderly and non-demented population.


Adjudin improves beta cell maturation, hepatic glucose uptake and glucose homeostasis.
Recovering functional beta cell mass is a promising approach for future diabetes therapies. The aim of the present study is to investigate the effects of adjudin, a small molecule identified in a beta cell screen using zebrafish, on pancreatic beta cells and diabetes conditions in mice and human spheroids. In zebrafish, insulin expression was examined by bioluminescence and quantitative real-time PCR (qPCR), glucose levels were examined by direct measurements and distribution using a fluorescent glucose analogue, and calcium activity in beta cells was analysed by in vivo live imaging. Pancreatic islets of wild-type postnatal day 0 (P0) and 3-month-old (adult) mice, as well as adult db/db mice (i.e. BKS(D)-Leprdb/JOrlRj), were cultured in vitro and analysed by qPCR, glucose stimulated insulin secretion and whole mount staining. RNA-seq was performed for islets of P0 and db/db mice. For in vivo assessment, db/db mice were treated with adjudin and subjected to analysis of metabolic variables and islet cells. Glucose consumption was examined in primary human hepatocyte spheroids. Adjudin treatment increased insulin expression and calcium response to glucose in beta cells and decreased glucose levels after beta cell ablation in zebrafish. Adjudin led to improved beta cell function, decreased beta cell proliferation and glucose responsive insulin secretion by decreasing basal insulin secretion in in vitro cultured newborn mouse islets. RNA-seq of P0 islets indicated that adjudin treatment resulted in increased glucose metabolism and mitochondrial function, as well as downstream signalling pathways involved in insulin secretion. In islets from db/db mice cultured in vitro, adjudin treatment strengthened beta cell identity and insulin secretion. RNA-seq of db/db islets indicated adjudin-upregulated genes associated with insulin secretion, membrane ion channel activity and exocytosis. Moreover, adjudin promoted glucose uptake in the liver of zebrafish in an insulin-independent manner, and similarly promoted glucose consumption in primary human hepatocyte spheroids with insulin resistance. In vivo studies using db/db mice revealed reduced nonfasting blood glucose, improved glucose tolerance and strengthened beta cell identity after adjudin treatment. Adjudin promoted functional maturation of immature islets, improved function of dysfunctional islets, stimulated glucose uptake in liver and improved glucose homeostasis in db/db mice. Thus, the multifunctional drug adjudin, previously studied in various contexts and conditions, also shows promise in the management of diabetic states. Raw and processed RNA-seq data for this study have been deposited in the Gene Expression Omnibus under accession number GSE235398 ( https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE235398 ).