Radiography of perforating centerfire rifle wounds of the trunk.
All deaths resulting from perforating centerfire rifle wounds of the chest and abdomen, investigated by the Office of the Chief Medical Examiner for the Province of Alberta from 1988 to 1995. were reviewed retrospectively to determine whether the radiographic distribution of bullet fragments in such cases is a useful predictor of bullet trajectory. Study cases were limited to single gunshot wounds without surgical intervention or intermediate targets, and for which adequate radiography was available. Three pathologists individually viewed the radiographs on two separate occasions; wound locations were provided for the second viewing (Group 2). Differences in opinion regarding direction of fire were resolved by consensus review. A trauma radiologist independently made two sets of interpretations in the same way. Comparisons of these groups of interpretations were made with the actual bullet direction determined at autopsy. Of 21 cases included in the study, only three (14.3%) did not require consensus resolution in either group. Accuracy of pathologists' interpretation improved from 38.1% (8/21) to 76.2% (16/21) with provision of wound locations (p = 0.012). The radiologist achieved similar improvement, from 28.6% (6/21) to 47.6% (10/21). The rate of agreement between radiologist and pathologists increased from 42.9% (9/21) to 61.9% (13/21) between Groups 1 and 2. Both the pathologists and radiologist interpreted several cases the same way in both groups; of those cases interpreted differently, the second interpretation was occasionally incorrect after correct interpretation in Group 1. We conclude that bullet direction for perforating centerfire rifle wounds cannot be accurately determined from postmortem radiographs. When wound location is known, the ability to predict bullet direction improves but is still subject to error, including a lack of consistency between observers.


Nonpathological phalangeal fractures in cattle: 17 cases (2004-2020).
To characterize clinical findings and outcomes for cattle with nonpathological phalangeal fractures. 17 cattle with nonpathological phalangeal fractures. Medical records of a teaching hospital were reviewed to identify cattle treated for nonpathological phalangeal fracture between May 2004 and May 2020. Information extracted from the records of study-eligible animals included signalment, history, clinical and diagnostic imaging findings, treatment, and survival to hospital discharge. Long-term outcome was assessed by telephone communication with owners. 9 bulls and 8 heifers or cows (age range, 1 to 7 years) of various breeds and uses were evaluated. Five of the 9 bulls were bucking stock, which were overrepresented in the study population. Seven animals had 8 distal phalanx fractures; 10 animals had 11 proximal phalanx fractures. Eight animals were treated by application of a hoof block on the unaffected adjacent digit, 7 animals were treated with a distal limb (n = 6) or transfixation pin (1) cast in addition to a hoof block, 1 animal was treated with a hoof trim to elevate and alleviate weight bearing on the affected digit, and 1 animal was euthanized immediately after diagnosis. Sixteen animals survived to hospital discharge. Follow-up was obtained for 12 animals, of which 9 returned to functionable use and 3 were culled. Results suggested cattle with a nonpathological phalangeal fracture have a favorable prognosis for return to function following application of a hoof block to the unaffected adjacent digit with or without a distal limb cast.


Informal caregiving as a risk factor for type 2 diabetes in individuals with favourable and unfavourable psychosocial work environments: A longitudinal multi-cohort study.
To examine whether informal caregiving is associated with increased risk of type 2 diabetes (T2D), and whether job strain and social support at work modify the association. Individual participant's data were pooled from three cohort studies-the French GAZEL study, the Swedish Longitudinal Occupational Survey of Health (SLOSH) and the British Whitehall II study-a total of 21,243 study subjects. Informal caregiving was defined as unpaid care for a closely related person. Job strain was assessed using the demand-control model, and questions on co-worker and supervisor support were combined in a measure of social support at work. Incident T2D was ascertained using registry-based, clinically assessed and self-reported data. A total of 1058 participants developed T2D during the up to 10 years of follow-up. Neither informal caregiving (OR: 1.09, 95% CI: 0.92-1.30) nor high job strain (OR: 1.04, 95% CI: 0.86-1.26) were associated with T2D risk, whereas low social support at work was a risk factor for T2D (OR: 1.18, 95% CI: 1.02-1.37). Also, informal caregivers who were also exposed to low social support at work were at higher risk of T2D (OR: 1.40, 95% CI: 1.08-1.82) compared with those who were not informal caregivers and had high social support at work (multiplicative test for interaction, P=0.04; additive test for interaction, synergy index=10). Informal caregiving was not independently associated with T2D risk. However, low social support at work was a risk factor, and informal caregivers with low social support at work had even higher risks of T2D.


Measuring Absolute RNA Copy Numbers at High Temporal Resolution Reveals Transcriptome Kinetics in Development.
Transcript regulation is essential for cell function, and misregulation can lead to disease. Despite technologies to survey the transcriptome, we lack a comprehensive understanding of transcript kinetics, which limits quantitative biology. This is an acute challenge in embryonic development, where rapid changes in gene expression dictate cell fate decisions. By ultra-high-frequency sampling of Xenopus embryos and absolute normalization of sequence reads, we present smooth gene expression trajectories in absolute transcript numbers. During a developmental period approximating the first 8 weeks of human gestation, transcript kinetics vary by eight orders of magnitude. Ordering genes by expression dynamics, we find that "temporal synexpression" predicts common gene function. Remarkably, a single parameter, the characteristic timescale, can classify transcript kinetics globally and distinguish genes regulating development from those involved in cellular metabolism. Overall, our analysis provides unprecedented insight into the reorganization of maternal and embryonic transcripts and redefines our ability to perform quantitative biology.


Genetic variances of SNP loci for milk yield in dairy cattle.
Regression coefficients and genetic variances for 40,890 single nucleotide polymorphisms (SNPs) for milk yield were calculated using mixed model equations, with deregressed proof (DRP) as the dependent variable. Bulls were genotyped using the Illumina BovineSNP50 v2 BeadChip and SNPs were edited according the minor allele frequency (MAF) and high incidence of missing genotype. Evaluation was conducted in two rounds. In the preliminary round, the direct genetic values (DGVs) of all genotyped bulls (2,904) were computed and the absolute difference between the DGV and the input DRP of each bull was investigated. Bulls with an absolute difference greater than the mean absolute difference plus two standard deviations were eliminated from the data set prior to the final analysis (2,766 bulls remaining). SNP regression coefficients from the final analysis had a mean absolute value of 0.506 kg and a standard deviation of 0.409 kg. The SNP with the highest regression coefficient and genetic variance was ARSBFGLNGS4939 on chromosome 14. This SNP is located within the gene DGAT1 (diacylglycerol O-acyltransferase 1). Other SNPs with high regression coefficients and genetic variance are localised in proximity to DGAT1. The mean genetic variance of an individual SNP was 0.170, with a standard deviation of 0.384 and a mean heterozygosity of 0.372. The sum of genetic variances of all SNPs was only 6,968.8, probably because of the existence of genetic covariances between loci. The largest sum of genetic variances was on chromosome 14 (498.4, 7.15 % of the total). After the final analysis, the correlation between the DGV and the input DRP was 0.951 for all bulls. The variance of the predicted DGV was 98.11 % of the variance of the input estimated breeding value (EBV) and 63.65 % of the variance of the DRP.


Accuracy in the diagnosis of short-segment Barrett's esophagus: the role of endoscopic experience.
The diagnosis of Barrett's esophagus is based on histologic demonstration of specialized intestinal metaplasia. Experience may be important in the endoscopic recognition of Barrett's esophagus, including in regard to appropriate procurement of biopsy specimens. The aim of this study was to assess factors that may influence accuracy in the diagnosis of short-segment Barrett's esophagus (SSB). Endoscopy reports pertaining to procedures performed over a 1-year period that included esophageal biopsies because of suspected intestinal metaplasia were reviewed. Barrett's epithelium involving less than 2 cm of the distal esophagus was considered SSB; greater than 2 cm was considered long-segment (LSB). Endoscopists were regarded as "more experienced" if they had completed training more than 5 years earlier and "less experienced" if the time elapsed since the completion of training was less than 5 years. More and less experienced endoscopists both obtained esophageal biopsy specimens because of suspected Barrett's esophagus at the same rate (14%). Length of suspected Barrett's epithelium was not predicted by symptoms or demographic data. Endoscopically, patients with SSB had significantly fewer (64.2% vs. 90.8%) and smaller (2.9 +/- 0.1 vs. 3.5 +/- 0.2 cm) hiatal hernias compared with those with LSB (p < 0.05). Suspected SSB was histologically confirmed in 38.4% (True SSB), whereas LSB was confirmed in 75% (True SSB) (p < 0.05). More experienced endoscopists were significantly more likely to obtain histologic confirmation of SSB than less experienced endoscopists (48.6% vs. 29.5%; p = 0.02, nominal significance from univariate hypothesis testing; correction for multiple testing of data removed significance at the p < 0.05 level; OR = 2.26). With greater experience, an endoscopist is more likely to diagnose SSB. This may be due to more accurate procurement of adequate tissue samples, which thereby results in a greater yield of histopathologic diagnoses.


A variant TMPRSS2 isoform and ERG fusion product in prostate cancer with implications for molecular diagnosis.
Prostate cancer is the most commonly diagnosed cancer among men in the United States. Recently, fusion of TMPRSS2 with ETS family oncogenic transcription factors has been identified as a common molecular alteration in prostate cancer, where most often the rearrangement places ERG under the androgen-regulated transcriptional control of TMPRSS2. Here, we carried out rapid amplification of cDNA ends (RACE) on a prostate cancer specimen carrying an atypical aberration discovered by array-based comparative genomic hybridization (array CGH), suggesting an alternative fusion partner of ERG. We identified novel transcribed sequences fused to ERG, mapping 4 kb upstream of the TMPRSS2 start site. The sequences derive from an apparent second TMPRSS2 isoform, which we found also expressed in some prostate tumors, suggesting similar androgen-regulated control. In a reverse transcription-polymerase chain reaction (RT-PCR)-based survey of 63 prostate tumor specimens (54 primary and nine lymph node metastases), 44 (70%) cases expressed either the known or novel variant TMPRSS2-ERG fusion, 28 (44%) expressed both, 10 (16%) expressed only the known, and notably six (10%) expressed only the variant isoform fusion. In this specimen set, the presence of a TMPRSS2-ERG fusion showed no statistical association with tumor stage, Gleason grade or recurrence-free survival. Nonetheless, the discovery of a novel variant TMPRSS2 isoform-ERG fusion adds to the characterization of ETS-family rearrangements in prostate cancer, and has important implications for the accurate molecular diagnosis of TMPRSS2-ETS fusions.


Improved single particle localization accuracy with dual objective multifocal plane microscopy.
In single particle imaging applications, the number of photons detected from the fluorescent label plays a crucial role in the quantitative analysis of the acquired data. For example, in tracking experiments the localization accuracy of the labeled entity can be improved by collecting more photons from the labeled entity. Here, we report the development of dual objective multifocal plane microscopy (dMUM) for single particle studies. The new microscope configuration uses two opposing objective lenses, where one of the objectives is in an inverted position and the other objective is in an upright position. We show that dMUM has a higher photon collection efficiency when compared to standard microscopes. We demonstrate that fluorescent labels can be localized with better accuracy in 2D and 3D when imaged through dMUM than when imaged through a standard microscope. Analytical tools are introduced to estimate the nanoprobe location from dMUM images and to characterize the accuracy with which they can be determined.


The Expanding Field of Secondary Antibody Deficiency: Causes, Diagnosis, and Management.
Antibody deficiency or hypogammaglobulinemia can have primary or secondary etiologies. Primary antibody deficiency (PAD) is the result of intrinsic genetic defects, whereas secondary antibody deficiency may arise as a consequence of underlying conditions or medication use. On a global level, malnutrition, HIV, and malaria are major causes of secondary immunodeficiency. In this review we consider secondary antibody deficiency, for which common causes include hematological malignancies, such as chronic lymphocytic leukemia or multiple myeloma, and their treatment, protein-losing states, and side effects of a number of immunosuppressive agents and procedures involved in solid organ transplantation. Secondary antibody deficiency is not only much more common than PAD, but is also being increasingly recognized with the wider and more prolonged use of a growing list of agents targeting B cells. SAD may thus present to a broad range of specialties and is associated with an increased risk of infection. Early diagnosis and intervention is key to avoiding morbidity and mortality. Optimizing treatment requires careful clinical and laboratory assessment and may involve close monitoring of risk parameters, vaccination, antibiotic strategies, and in some patients, immunoglobulin replacement therapy (IgRT). This review discusses the rapidly evolving list of underlying causes of secondary antibody deficiency, specifically focusing on therapies targeting B cells, alongside recent advances in screening, biomarkers of risk for the development of secondary antibody deficiency, diagnosis, monitoring, and management.


Simple heterodyne laser interferometer with subnanometer periodic errors.
We describe a simple heterodyne laser interferometer that has subnanometer periodic errors and is applicable to industrial fields. Two spatially separated beams can reduce the periodic errors, and the use of a right-angle prism makes the optical configuration much simpler than previous interferometers. Moreover, the optical resolution can be enhanced by a factor of 2, because the phase change direction is opposite between reference and measurement signals. Experiments have demonstrated the periodic errors are less than 0.15 nm owing to the frequency mixing of the optical source. The improvements for reducing the frequency mixing of the optical system are also discussed.


Racial and sexual identity-related maltreatment among minority YMSM: prevalence, perceptions, and the association with emotional distress.
Bullying is a form of violence characterized as an aggressive behavior that is unprovoked and intended to cause harm. Prior studies have found that lesbian, gay, bisexual, and transgender (LGBT) youth experience high levels of bullying related to their sexuality and this harassment can lead to engagement in risk behaviors, depression, and suicide. Ethnic/racial minority young men who have sex with men (YMSM) may experience dual levels of stigma and maltreatment due to both their sexuality and their race. The aim of the current study was to assess the prevalence and perceptions of racial and sexual identity-based abuse among a sample of minority YMSM, and whether this maltreatment plays a role in the emotional distress of these youth. We found that overall 36% and 85% of participants experienced racial and sexuality-related bullying, respectively. There was a significant association between experiencing a high level of sexuality-related bullying and depressive symptomatology (p=0.03), having attempted suicide (p=0.03), and reporting parental abuse (p=0.05). We found no association between racial bullying and suicide attempts. In a multivariable logistic regression model, experiencing any racial bullying and high sexuality-related bullying were significant predictors of having a CES-D score ≥16; adjusted odds ratio (OR) 1.83 and 2.29, respectively. These findings contribute to the existing literature regarding the negative experiences and daily stressors facing LGBT youth with regard to both their minority status and LGBT identities. Future interventions for racial/ethnic minority YMSM should provide assistance to achieve a positive view of self that encompasses both their racial and sexual identities.


Disease associated with equine coronavirus infection and high case fatality rate.
Equine coronavirus (ECoV) is associated with clinical disease in adult horses. Outbreaks are associated with a low case fatality rate and a small number of animals with signs of encephalopathic disease are described. The aim of this study is to describe the epidemiological and clinical features of two outbreaks of ECoV infection that were associated with an high case fatality rate. 14 miniature horses and 1 miniature donkey testing fecal positive for ECoV from two related disease outbreaks. Retrospective study describing the epidemiological findings, clinicopathological findings, and fecal viral load from affected horses. In EcoV positive horses, 27% (4/15) of the animals died or were euthanized. Severe hyperammonemia (677 μmol/L, reference range ≤ 60 μmol/L) was identified in one animal with signs of encephalopathic disease that subsequently died. Fecal viral load (ECoV genome equivalents per gram of feces) was significantly higher in the nonsurvivors compared to animals that survived (P = .02). Equine coronavirus had a higher case fatality rate in this group of miniature horses than previously reported in other outbreaks of varying breeds. Hyperammonemia could contribute to signs of encephalopathic disease, and the fecal viral load might be of prognostic value in affected horses.


Layer-dependent electronic properties of phosphorene-like materials and phosphorene-based van der Waals heterostructures.
Black phosphorus is a layered semiconducting allotrope of phosphorus with high carrier mobility. Its monolayer form, phosphorene, is an extremely fashionable two-dimensional material which has promising potential in transistors, optoelectronics and electronics. However, phosphorene-like analogues, especially phosphorene-based heterostructures and their layer-controlled electronic properties, are rarely systematically investigated. In this paper, the layer-dependent structural and electronic properties of phosphorene-like materials, i.e., mono- and few-layer MXs (M = Sn, Ge; X = S, Se), are first studied via first-principles calculations, and then the band edge position of these MXs as well as mono- and few-layer phosphorene are aligned. It is revealed that van der Waals heterostructures with a Moiré superstructure formed by mutual coupling among MXs and among MXs and few-layer phosphorene are able to show type-I or type-II characteristics and a I-II or II-I transition can be induced by adjusting the number of layers. Our work is expected to yield a new family of phosphorene-based semiconductor heterostructures with tunable electronic properties through altering the number of layers of the composite.


Effects of dapagliflozin on blood pressure in hypertensive diabetic patients on renin-angiotensin system blockade.
Hypertension and type 2 diabetes mellitus (T2DM) are risk factors for cardiovascular disease. Dapagliflozin improves glycemic control and systolic blood pressure (SBP) in T2DM patients. This double-blind phase III study evaluated the effects of dapagliflozin on glycemic control and blood pressure in patients with inadequately controlled T2DM and hypertension, despite ongoing therapy with a renin-angiotensin system blocker. Patients were randomized to receive dapagliflozin 10 mg (n = 302) or placebo (n = 311) once daily for 12 weeks. Endpoints were change from baseline to week 12 in seated SBP and glycosylated hemoglobin (HbA1c); longitudinal repeated-measures analysis was performed. Additional endpoints included other hemodynamic measures, serum uric acid, fasting plasma glucose, body weight, blood lipids and heart rate. After 12 weeks, dapagliflozin-treated versus placebo-treated patients showed significant reductions in HbA1c (-0.6% vs -0.1%, p < 0.0001), mean seated SBP (-10.4 vs -7.3 mmHg, p = 0.0010) and mean 24 h ambulatory SBP (-9.6 vs -6.7 mmHg, p = 0.0043). Dapagliflozin also reduced body weight compared with placebo (-1.0 vs -0.3 kg). Dapagliflozin was well tolerated, with adverse events consistent with previous studies. Dapagliflozin improved glycemic control, and reduced SBP as well as body weight in patients with poorly controlled T2DM and hypertension.


Catch Handling and the Possible Migration of Anisakis Larvae in Herring, Clupea harengus.
The behavior of larvae of Anisakis sp. in North Sea herring (Clupea harengus) after capture was examined by a sensitive quantitative method (digestion). In one winter and one summer experiment, the mean numbers of worms in freshly caught herring were 0.06 and 0.09 (in double fillets), 0.19 and 0.24 (in double belly flaps), and 10.4 and 7.8 larvae (in viscera), respectively. In each experiment herrings were stored ungutted for up to 5 1/2 d on ice (0°C), in refrigerated/chilled sea water (-10°C), or in warm sea water (10°C), but no changes in the numbers of Anisakis larvae in the belly flaps and the fillets could be demonstrated. The present results show that Anisakis larvae are present in the flesh of herring already at capture, but no significant postmortem migration into the flesh could be demonstrated during storage. Thus, immediately gutting on board cannot eliminate or even reduce the risk from eating raw or inadequately processed herring.


Probing ion current in solid-electrolytes at the meso- and nanoscale.
We present experimental approaches to probe the ionic conductivity of solid electrolytes at the meso- and nanoscales. Silica ionogel based electrolytes have emerged as an important class of solid electrolytes because they maintain both fluidic and high-conductivity states at the nanoscale, but at the macroscale they are basically solid. Single mesopores in polymer films are shown to serve as templates for cast ionogels. The ionic conductivity of the ionogels was probed by two experimental approaches. In the first approach, the single-pore/ionogel membranes were placed between two chambers of a conductivity cell, in a set-up similar to that used for investigating liquid electrolytes. The second approach involved depositing contacts directly onto the membrane and measuring conductivity without the bulk solution present. Ionic conductivity determined by the two methods was in excellent agreement with macroscopic measurements, which suggested that the electrochemical properties of ionogel based electrolytes are preserved at the mesoscale, and ionogels can be useful in designing meso-scaled energy-storage devices.


Effect of obesity on survival of women with epithelial ovarian cancer: a systematic review and meta-analysis of observational studies.
Although obesity is shown to be a risk factor for epithelial ovarian cancer, its role as a prognostic factor has been remained inconclusive. In this study, available evidences on this matter to date have been assembled for a meta-analysis to determine the effect of obesity on the survival of patients with epithelial ovarian cancer. Eligible studies published up to December 2010 were searched using MEDLINE (PubMed), EMBASE, and Cochrane Central Register of Controlled Trials, and manual review of relevant bibliography to look for additional studies was done. Adjusted hazard ratios (HRs) from individual studies were pooled using a random-effects model. Ten cohort studies of 331 screened articles were included in the final analysis. The meta-analysis showed overweight or obesity at early adulthood to be associated with higher mortality among patients with ovarian cancer (HR, 1.60; 95% confidence interval, 1.10-2.34). Among patients with advanced ovarian cancer, premorbid obesity was associated with worse prognosis (HR, 1.45; 95% confidence interval, 1.09-1.93). However, there was no significant relationship between prognosis and obesity around the time of diagnosis. This study suggests a possible relationship between obesity at early adulthood and higher mortality among patients with ovarian cancer. Further studies are needed to elucidate the harmful effect of obesity on the survival of patients with ovarian cancer.


Reversal of the posttranslational modification on Chlamydomonas flagellar alpha-tubulin occurs during flagellar resorption.
We previously have shown that a posttranslational modification of alpha-tubulin takes place in the flagellum during Chlamydomonas flagellar assembly (L'Hernault, S. W., and J. L. Rosenbaum, 1983, J. Cell Biol., 97:258-263). In this report, we show that the posttranslationally modified alpha-3 tubulin is changed back to its unmodified alpha-1 precursor form during the microtubular disassembly that takes place during flagellar resorption. These data indicate that the addition and removal of a posttranslational modification on alpha-tubulin might be a control step in the assembly and disassembly of flagella.


[Trial end-point in chronic obstructive pulmonary disease (COPD): minimal clinically important difference].
The concept of the minimal clinically important difference (MCID) is intended to provide a measure of relevance for a statistically applied in patients with COPD. Clinically important differences are those differences relevant to the individual patient and important to the patient's life. However, people's difference in a diagnostic parameter perception of what is important vary. Furthermore, physicians may rate the significance of a particular marker and its difference which can be achieved by a pharmacological intervention differently from the patient. Thus, the major problem with defining an MCID for any measure is that the most important differences, which require the most subtle measures for an individual patient, are likely to have the least general application. Conversely, measures that can be generalised are unlikely to have much individual importance and will be very crude tools for an individual assessment. In medical trials both, statistical rigor and clinical relevance are generally required, and MCID is without doubt a key application tool defining treatment success or treatment failure. This paper gives an update on the concept of a minimal important difference of most relevant parameters in COPD treatment.


COVID-19: Rational discovery of the therapeutic potential of Melatonin as a SARS-CoV-2 main Protease Inhibitor.
The SARS-CoV-2 spread quickly across the globe. The World Health Organization (WHO) on March 11 declared COVID-19 a pandemic. The mortality rate, hospital disorders and incalculable economic and social damages, besides the unproven efficacy of the treatments evaluated against COVID-19, raised the need for immediate control of this disease. Therefore, the current study employed in silico tools to rationally identify new possible SARS-CoV-2 main protease (Mpro) inhibitors. That is an enzyme conserved among the coronavirus species; hence, the identification of an Mpro inhibitor is to make it a broad-spectrum drug. Molecular docking studies described the binding sites and the interaction energies of 74 Mpro-ligand complexes deposited in the Protein Data Bank (PDB). A structural similarity screening was carried out in order to identify possible Mpro ligands that show additional pharmacological properties against COVID-19. We identified 59 hit compounds and among them, melatonin stood out due to its prominent immunomodulatory and anti-inflammatory activities; it can reduce oxidative stress, defence cell mobility and efficiently combat the cytokine storm and sepsis. In addition, melatonin is an inhibitor of calmodulin, an essential intracellular component to maintain angiotensin-converting enzyme 2 (ACE-2) on the cell surface. Interestingly, one of the most promising hits in our docking study was melatonin. It revealed better interaction energy with Mpro compared to ligands in complexes from PDB. Consequently, melatonin can have response potential in early stages for its possible effects on ACE-2 and Mpro, although it is also promising in more severe stages of the disease for its action against hyper-inflammation. These results definitely do not confirm antiviral activity, but can rather be used as a basis for further preclinical and clinical trials.


Association of Autologous Tumor Lysate-Loaded Dendritic Cell Vaccination With Extension of Survival Among Patients With Newly Diagnosed and Recurrent Glioblastoma: A Phase 3 Prospective Externally Controlled Cohort Trial.
Glioblastoma is the most lethal primary brain cancer. Clinical outcomes for glioblastoma remain poor, and new treatments are needed. To investigate whether adding autologous tumor lysate-loaded dendritic cell vaccine (DCVax-L) to standard of care (SOC) extends survival among patients with glioblastoma. This phase 3, prospective, externally controlled nonrandomized trial compared overall survival (OS) in patients with newly diagnosed glioblastoma (nGBM) and recurrent glioblastoma (rGBM) treated with DCVax-L plus SOC vs contemporaneous matched external control patients treated with SOC. This international, multicenter trial was conducted at 94 sites in 4 countries from August 2007 to November 2015. Data analysis was conducted from October 2020 to September 2021. The active treatment was DCVax-L plus SOC temozolomide. The nGBM external control patients received SOC temozolomide and placebo; the rGBM external controls received approved rGBM therapies. The primary and secondary end points compared overall survival (OS) in nGBM and rGBM, respectively, with contemporaneous matched external control populations from the control groups of other formal randomized clinical trials. A total of 331 patients were enrolled in the trial, with 232 randomized to the DCVax-L group and 99 to the placebo group. Median OS (mOS) for the 232 patients with nGBM receiving DCVax-L was 19.3 (95% CI, 17.5-21.3) months from randomization (22.4 months from surgery) vs 16.5 (95% CI, 16.0-17.5) months from randomization in control patients (HR = 0.80; 98% CI, 0.00-0.94; P = .002). Survival at 48 months from randomization was 15.7% vs 9.9%, and at 60 months, it was 13.0% vs 5.7%. For 64 patients with rGBM receiving DCVax-L, mOS was 13.2 (95% CI, 9.7-16.8) months from relapse vs 7.8 (95% CI, 7.2-8.2) months among control patients (HR, 0.58; 98% CI, 0.00-0.76; P < .001). Survival at 24 and 30 months after recurrence was 20.7% vs 9.6% and 11.1% vs 5.1%, respectively. Survival was improved in patients with nGBM with methylated MGMT receiving DCVax-L compared with external control patients (HR, 0.74; 98% CI, 0.55-1.00; P = .03). In this study, adding DCVax-L to SOC resulted in clinically meaningful and statistically significant extension of survival for patients with both nGBM and rGBM compared with contemporaneous, matched external controls who received SOC alone. ClinicalTrials.gov Identifier: NCT00045968.


RNAi screening: new approaches, understandings, and organisms.
RNA interference (RNAi) leads to sequence-specific knockdown of gene function. The approach can be used in large-scale screens to interrogate function in various model organisms and an increasing number of other species. Genome-scale RNAi screens are routinely performed in cultured or primary cells or in vivo in organisms such as C. elegans. High-throughput RNAi screening is benefitting from the development of sophisticated new instrumentation and software tools for collecting and analyzing data, including high-content image data. The results of large-scale RNAi screens have already proved useful, leading to new understandings of gene function relevant to topics such as infection, cancer, obesity, and aging. Nevertheless, important caveats apply and should be taken into consideration when developing or interpreting RNAi screens. Some level of false discovery is inherent to high-throughput approaches and specific to RNAi screens, false discovery due to off-target effects (OTEs) of RNAi reagents remains a problem. The need to improve our ability to use RNAi to elucidate gene function at large scale and in additional systems continues to be addressed through improved RNAi library design, development of innovative computational and analysis tools and other approaches.


Changes in Cardiovascular Biomarkers With Breast Cancer Therapy and Associations With Cardiac Dysfunction.
Background We examined the longitudinal associations between changes in cardiovascular biomarkers and cancer therapy-related cardiac dysfunction (CTRCD) in patients with breast cancer treated with cardotoxic cancer therapy. Methods and Results Repeated measures of high-sensitivity cardiac troponin T (hs-cTnT), NT-proBNP (N-terminal pro-B-type natriuretic peptide), myeloperoxidase, placental growth factor, and growth differentiation factor 15 were assessed longitudinally in a prospective cohort of 323 patients treated with anthracyclines and/or trastuzumab followed over a maximum of 3.7 years with serial echocardiograms. CTRCD was defined as a ≥10% decline in left ventricular ejection fraction to a value <50%. Associations between changes in biomarkers and left ventricular ejection fraction were evaluated in repeated-measures linear regression models. Cox regression models assessed the associations between biomarkers and CTRCD. Early increases in all biomarkers occurred with anthracycline-based regimens. hs-cTnT levels >14 ng/L at anthracycline completion were associated with a 2-fold increased CTRCD risk (hazard ratio, 2.01; 95% CI, 1.00-4.06). There was a modest association between changes in NT-proBNP and left ventricular ejection fraction in the overall cohort; this was most pronounced with sequential anthracycline and trastuzumab (1.1% left ventricular ejection fraction decline [95% CI, -1.8 to -0.4] with each NT-proBNP doubling). Increases in NT-proBNP were also associated with CTRCD (hazard ratio per doubling, 1.56; 95% CI, 1.32-1.84). Increases in myeloperoxidase were associated with CTRCD in patients who received sequential anthracycline and trastuzumab (hazard ratio per doubling, 1.28; 95% CI, 1.04-1.58). Conclusions Cardiovascular biomarkers may play an important role in CTRCD risk prediction in patients with breast cancer who receive cardiotoxic cancer therapy, particularly in those treated with sequential anthracycline and trastuzumab therapy. Clinical Trial Registration URL: https://www.clinicaltrials.gov/. Unique identifier: NCT01173341.


Influence of rye flour enzymatic biotransformation on the antioxidant capacity and transepithelial transport of phenolic acids.
Phenolic acids have been reported to play a role on the antioxidant activity and other important biological activities. However, as most polyphenolics in food products are either bound to cellular matrices or present as free polymeric forms, the way they are absorbed has not been totally clear until now. Hydrolytic enzymes may act to increase functionalities in polyphenolic-rich foods, enhancing the bioaccessibility of phenolic compounds and minerals from whole grains. The aim of this study was to evaluate the action of tannin acyl hydrolase (tannase) on the total phenols, phenolic acid profile, antioxidant capacity and in vitro bioaccessibility of phenolic acids found in whole rye flour (RF). Besides increasing total phenols and the antioxidant capacity, tannase treatment increased the amounts of ferulic, sinapic and vanillic acids identified in RF, evidencing a new type of feruloyl esterase catalytic action of tannase. Vanillic and sinapic acids in tannase-treated whole rye flour (RFT) were higher than RF after in vitro gastrointestinal digestion, and higher amounts of transported vanillic acid through the Caco-2 monolayer were detected in RFT. However, the bioaccessibility and the transport efficiency of RF phenolic acids were higher than RFT. Underutilized crops like rye and rye-derived products may be an important source of phenolic acids. The tannase biotransformation, even influencing the total phenolics and antioxidant capacity of RF, did not increase the bioaccessibility of phenolic acids under the experimental conditions of this study.


"It pains me because as a woman you have to breastfeed your baby": decision-making about infant feeding among African women living with HIV in the UK.
UK guidance advises HIV-positive women to abstain from breast feeding. Although this eliminates the risk of postnatal vertical transmission of HIV, the impact of replacement feeding on mothers is often overlooked. This qualitative study examines, for the first time in the UK, decision-making about infant feeding among African women living with HIV. Between 2010 and 2011, we conducted semistructured interviews with 23 HIV-positive African women who were pregnant or had recently given birth. We recruited participants from three HIV antenatal clinics in London. Women highlighted the cultural importance of breast feeding in African communities and the social pressure to breast feed, also describing fears that replacement feeding would signify their HIV status. Participants had significant concerns about physical and psychological effects of replacement feeding on their child and felt their identity as good mothers was compromised by not breast feeding. However, almost all chose to refrain from breast feeding, driven by the desire to minimise vertical transmission risk. Participants' resilience was strengthened by financial assistance with replacement feeding, examples of healthy formula-fed children and support from partners, family, peers and professionals. The decision to avoid breast feeding came at considerable emotional cost to participants. Professionals should be aware of the difficulties encountered by HIV-positive women in refraining from breast feeding, especially those from migrant African communities where breast feeding is culturally normative. Appropriate financial and emotional support increases women's capacity to adhere to their infant-feeding decisions and may reduce the emotional impact.


Cranberry intervention in patients with prostate cancer prior to radical prostatectomy. Clinical, pathological and laboratory findings.
Recently, we described an inverse association between cranberry supplementation and serum prostate specific antigen (PSA) in patients with negative biopsy for prostate cancer (PCa) and chronic nonbacterial prostatitis. This double blind placebo controlled study evaluates the effects of cranberry consumption on PSA values and other markers in men with PCa before radical prostatectomy. Prior to surgery, 64 patients with prostate cancer were randomized to a cranberry or placebo group. The cranberry group (n=32) received a mean 30 days of 1500 mg cranberry fruit powder. The control group (n=32) took a similar amount of placebo. Selected blood/urine markers as well as free and total phenolics in urine were measured at baseline and on the day of surgery in both groups. Prostate tissue markers were evaluated after surgery. The serum PSA significantly decreased by 22.5% in the cranberry arm (n=31, P<0.05). A trend to down-regulation of urinary beta-microseminoprotein (MSMB) and serum gamma-glutamyltranspeptidase, as well as upregulation of IGF-1 was found after cranberry supplementation. There were no changes in prostate tissue markers or, composition and concentration of phenolics in urine. Daily consumption of a powdered cranberry fruit lowered serum PSA in patients with prostate cancer. The whole fruit contains constituents that may regulate the expression of androgen-responsive genes.


Whole exome sequencing identifies a mutation for a novel form of corneal intraepithelial dyskeratosis.
Corneal intraepithelial dyskeratosis is an extremely rare condition. The classical form, affecting Native American Haliwa-Saponi tribe members, is called hereditary benign intraepithelial dyskeratosis (HBID). Herein, we present a new form of corneal intraepithelial dyskeratosis for which we identified the causative gene by using deep sequencing technology. A seven member Caucasian French family with two corneal intraepithelial dyskeratosis affected individuals (6-year-old proband and his mother) was ascertained. The proband presented with bilateral complete corneal opacification and dyskeratosis. Palmoplantar hyperkeratosis and laryngeal dyskeratosis were associated with the phenotype. Histopathology studies of cornea and vocal cord biopsies showed dyskeratotic keratinisation. Quantitative PCR ruled out 4q35 duplication, classically described in HBID cases. Next generation sequencing with mean coverage of 50× using the Illumina Hi Seq and whole exome capture processing was performed. Sequence reads were aligned, and screened for single nucleotide variants and insertion/deletion calls. In-house pipeline filtering analyses and comparisons with available databases were performed. A novel missense mutation M77T was discovered for the gene NLRP1 which maps to chromosome 17p13.2. This was a de novo mutation in the proband's mother, following segregation in the family, and not found in 738 control DNA samples. NLRP1 expression was determined in adult corneal epithelium. The amino acid change was found to destabilise significantly the protein structure. We describe a new corneal intraepithelial dyskeratosis and how we identified its causative gene. The NLRP1 gene product is implicated in inflammation, autoimmune disorders, and caspase mediated apoptosis. NLRP1 polymorphisms are associated with various diseases.


Prevalence and Clinical Correlates of Poststroke Behavioral Dysexecutive Syndrome.
Background Behavioral dysexecutive syndrome (BDES) is a common phenomenon following stroke. To date, research has focused mainly on individual behavioral symptoms rather than a more comprehensive characterization of goal-directed behavior in stroke survivors. This cross-sectional study evaluated the prevalence and clinical correlates of BDES in Hong Kong stroke survivors. Methods and Results A total of 369 stroke survivors were recruited from a regional hospital at 3 months after their index stroke. Patients' demographic and clinical characteristics were extracted from a comprehensive stroke database. BDES was measured with the Chinese version of the Dysexecutive Questionnaire. Four neurocognitive batteries assessed domains of cognitive executive functions. The prevalence of BDES 3 months poststroke was 18.7%. At that time point, the Hospital Anxiety Depression Scale and Mini-Mental State Examination scores and the presence of depression were significant predictors of BDES in a multivariate logistic regression analysis. These parameters remained significant predictors of the Dysexecutive Questionnaire score in a linear stepwise regression analysis and together accounted for 28.5% of the variance. Current depression was predictive of the Dysexecutive Questionnaire score in patients with BDES, with a variance of 9.7%. Furthermore, compared with the non-BDES group, patients with BDES exhibited poor performance-based executive function in the Chinese version of the Frontal Assessment Battery and color trails, arrow, and category fluency tests. Conclusions Symptoms of anxiety, current depression, and global cognitive function may be independent predictors of the presence and severity of BDES 3 months poststroke. Stroke survivors with BDES exhibit poor executive functioning, including goal maintenance and semantic memory.


Unexpected four-membered over six-membered ring formation during the synthesis of azaheterocyclic phosphonates: experimental and theoretical evaluation.
The cyclization of functionalized aminophosphonates is studied on both experimental and theoretical grounds. In a recently described route to phosphono-beta-lactams [Stevens C. V.; Vekemans, W.; Moonen, K.; Rammeloo, T. Tetrahedron Lett. 2003, 44, 1619], it was found that starting from an ambident allylic anion only four-membered rings were formed without any trace of six-membered lactams. New anion trapping experiments revealed that the gamma-anion is highly reactive in intermolecular reactions. Ab initio calculations predict higher reaction barriers for the gamma-anion due to restricted rotation about the C-N bond and due to highly strained transition states during ring closure. The sodium or lithium counterion, explicit dimethyl ether solvent molecules, and bulk solvent effects were properly taken into account at various levels of theory.


Proteomic analysis of 14-3-3 zeta binding proteins in the mouse hippocampus.
14-3-3 proteins are ubiquitous molecular chaperones with important roles in brain development and neuronal function. Altered expression of 14-3-3 proteins has been reported in several neurologic and neurodegenerative disorders and identifying 14-3-3 binding proteins may provide important insights into the physiologic and pathophysiologic roles of these proteins. Particular interest has emerged on 14-3-3 zeta (ζ) in the setting of neuronal injury because reducing 14-3-3ζ levels triggers an endoplasmic reticulum stress-like response in neurons and increases vulnerability to excitotoxicity. Here we examined the subcellular distribution of 14-3-3ζ in the mouse hippocampus. We then used recombinant His-tagged 14-3-3ζ to pull-down interacting proteins from the mouse hippocampus followed by identification by liquid chromatography-mass spectrometry. 14-3-3ζ protein was present in the cytoplasm, microsomal compartment, nucleus and mitochondrial fractions of the mouse hippocampus. Recombinant 14-3-3ζ eluted 13 known 14-3-3 binding partners, including three other 14-3-3 isoforms, and 16 other proteins which have not previously been reported to bind 14-3-3ζ. The present study identifies potentially novel 14-3-3ζ binding proteins and contributes to defining the 14-3-3ζ interactome in the mouse brain.


Optical properties of single diatom frustules revealed by confocal microspectroscopy.
Optical properties of single diatom frustule valves from two different Coscinodiscus species (C. wailesii and C. centralis) are studied by transmission confocal hyperspectral imaging and numerical calculations. Light convergence, concentration, and trapping effects are observed and depend on both the wavelength and the valve orientation. These effects seem to occur independently of the incident light angle. From our results, a wavelength-dependent multifocal lens behavior can be explained by light diffraction related to the radial symmetry of the multiscaled 3D nanostructure.


A nearly complete foot from Dikika, Ethiopia and its implications for the ontogeny and function of Australopithecus afarensis.
The functional and evolutionary implications of primitive retentions in early hominin feet have been under debate since the discovery of Australopithecus afarensis. Ontogeny can provide insight into adult phenotypes, but juvenile early hominin foot fossils are exceptionally rare. We analyze a nearly complete, 3.32-million-year-old juvenile foot of A. afarensis (DIK-1-1f). We show that juvenile A. afarensis individuals already had many of the bipedal features found in adult specimens. However, they also had medial cuneiform traits associated with increased hallucal mobility and a more gracile calcaneal tuber, which is unexpected on the basis of known adult morphologies. Selection for traits functionally associated with juvenile pedal grasping may provide a new perspective on their retention in the more terrestrial adult A. afarensis.


Detection of mosaicism at blastocyst stage with the use of high-resolution next-generation sequencing.
A significant proportion of human preimplantation embryos produced during the course of in vitro fertilization (IVF) treatments contain two or more cytogenetically distinct cell lines. This phenomenon, known as chromosomal mosaicism, can involve the presence of cells with different types of aneuploidy in the absence of any normal cells or a mixture of euploid and abnormal cells. Although a high prevalence of mosaicism at the cleavage and blastocyst stages has been appreciated for two decades, the precise frequency of the phenomenon and its consequences for embryo viability have been difficult to quantify. Recent advances in genetic technologies, such as high-resolution next-generation sequencing, have allowed mosaicism to be detected with much greater sensitivity than earlier methods. The application of these techniques to trophectoderm biopsies, taken from embryos before transfer to the uterus, has provided insight into the clinical impact of mosaicism. Data from recent studies show that blastocysts associated with mosaic trophectoderm biopsy specimens implant less often than embryos with a chromosomally normal biopsy. In addition, the mosaic embryos that succeed in establishing a pregnancy are at a significantly higher risk of miscarriage. Because mosaic embryos are less likely to produce a viable pregnancy than their euploid counterparts, we suggest that they are given a lower priority for transfer to the uterus. However, because these embryos can sometimes produce successful pregnancies, it is important that they can be considered for transfer in the absence of fully euploid embryos and after appropriate patient counseling. Unlike aneuploidy of meiotic origin, mosaicism, which is caused by mitotic errors occurring after fertilization, does not increase with advancing maternal age. There may, however, be clinical, treatment, or patient-related factors that contribute to the risk of mosaicism occurring. This review discusses the validation of methods that permit the detection of chromosomal mosaicism in IVF embryos and findings of clinical relevance.


Progressive arterial stiffening in kidney transplant recipients.
Pulse wave velocity (PWV) is a marker of arterial stiffness. Data from general population and end-stage renal disease patients suggest that PWV increases with age by about 0.1 m/s for each year of life and that PWV increase is steeper after the fifth decade. Recent follow-up studies showed decrease of arterial stiffness during the first year after kidney transplantation (KTx). This study aimed to investigate PWV change in kidney transplant recipients (KTR) during long-term follow-up. Carotid-femoral PWV was measured at baseline (PWV1) and then after at least 24 months (range 24-34, mean ±SD 28±3) of follow-up (PWV2) in 61 KTR aged 46±12 years. Baseline PWV measurements were performed at 36±27 months (range 4-121) after KTx. Clinical and laboratory data were also analyzed. GFR was calculated with MDRD formula (eGFR). ∆PWV was calculated as (PWV2-PWV1)/PWV1. Significant increase in PWV (9.1±1.7 vs. 9.8±2.0; p<0.002) and pulse pressure (PP) (50±11 vs. 53±14; p<0.05) was found during follow-up, but systolic, diastolic and mean arterial pressure did not change. eGFR did not change during follow-up (55±16 vs. 56±16 ml/min/1.73 m2). PWV increase was observed in patients aged ≥50 years (9.8±1.8 vs. 10.8±1.9; p<0.01), while not in younger patients (8.5±1.5 vs. 8.9±1.5; p=0.14). Significant positive correlation was found between ∆PWV and duration of the follow-up period. The study revealed progressive arterial stiffening in kidney transplant recipients during long-term follow-up. Further studies are necessary to explain the pathophysiology of this phenomenon.


Treatment of Chiari I malformation in patients with and without syringomyelia: a consecutive series of 66 cases.
The authors describe the results of performing a standard posterior craniovertebral decompression and placement, if indicated, of a syringosubarachnoid shunt for the treatment of patients with Chiari I malformation with and without syringomyelia. This is a retrospectively analyzed consecutive series of 66 patients (mean patient age 15 years, range 1-53 years). The uniform posterior craniovertebral decompression consisted of a small suboccipital craniectomy, a C-1 laminectomy, microsurgical reduction of the cerebellar tonsils, and dural closure with a synthetic dural graft to increase the cerebrospinal fluid space at the craniocervical junction. The presence of a large syrinx, with significant thinning of the spinal cord tissue and obliteration of the spinal subarachnoid space, particularly when combined with syrinx-related symptoms, was an indication for the placement of a syringosubarachnoid shunt. In 32 patients Chiari I malformation alone was present, and 34 in patients it was present in combination with syringomyelia. Clinical findings included pain, neurological deficits, and spinal deformity. The presence of syringomyelia was significantly associated with the presence of scoliosis (odds ratio 74.4 [95% confidence interval 8.894-622.4]). All patients underwent a posterior craniovertebral decompression procedure. In 22 of the 34 patients with syringomyelia a syringosubarachnoid shunt was also placed. The mean follow-up period was 24 months (range 3-95 months). Excellent outcome was achieved in 54 patients (82%) and good outcome in 12 (18%). In no patient were symptoms unchanged or worse at follow-up examination, including four patients who initially required a second operation for persistent syringomyelia. Pain was more likely to resolve than sensory and motor deficits after decompressive surgery. Radiological examination revealed normalization of tonsillar position in all patients. The syrinx had disappeared in 15 cases, was decreased in size in 17, and remained unchanged in two. Posterior craniovertebral decompression and selective placement of a syringosubarachnoid shunt in patients with Chiari I malformation and syringomyelia is an effective and safe treatment. Primary placement of a shunt in the presence of a sufficiently large syrinx appears to be beneficial. The question of if and when to place a shunt, however, requires further, preferably prospective, investigation.


Influence of poststimulation potentiation and heart rate on the fetal lamb heart.
Isolated cardiac muscle techniques and studies of the chronically instrumented fetal lamb heart were employed to evaluate the ability of fetal myocardium to exhibit poststimulation potentiation. Isometric tension development and the response to paired electrical stimulation were significantly reduced in isolated fetal ventricular myocardium when compared to the adult (P less than 0.001). As in the adult, increasing stimulation frequency raised fetal isometric tension via an increase in the rate of rise of tension development in the presence of reduction in time-to-peak tension. In seven fetal lambs the left ventricle was chronically instrumented with endocardial ultrasonic crystals and a high-fidelity micromanometer. After a 2-wk recovery period, heart rate was increased by atrial pacing from an average control level of 150 to 300 beats/min. Left ventricular dP/dt increased progressively and then fell beyond a heart rate of 270/min. When comparable pre- and immediate postpacing beats were analyzed, a step-wise increase in the velocity of left ventricular shortening and the mean rate of circumferential fiber shortening was observed in association with an increase in the extent of shortening. Thus, increases in the frequency of contraction exert a significant positive inotropic effect on the fetal heart.


A multigene typing system for human adenoviruses reveals a new genotype in a collection of Swedish clinical isolates.
Human adenoviruses (HAdVs) are common pathogens that can cause respiratory, gastrointestinal, urogenital, and ocular infections. They are divided into seven species containing 85 genotypes. Straightforward typing systems might help epidemiological investigations. As homologous recombination frequently shapes the evolution of HAdVs, information on a single gene is seldom sufficient to allow accurate and precise typing, and complete genome-based methods are recommended. Even so, complete genome analyses are not always easy to perform for practical reasons, and in such cases a multigene system can provide considerably more information about the strain under investigation than single-gene-based methods. Here we present a rapid, generic, multigene typing system for HAdVs based on three main deterministic regions of these viruses. Three PCR systems were used to amplify the genes encoding the DNA polymerase, the penton base hypervariable Arg-Gly-Asp-containing loop, and the hexon loop 1 (hypervariable region 1-6). Using this system, we typed 281 clinical isolates, detected members of six out of seven HAdV species (Human mastadenovirus A-F), and could also detect not only divergent strains of established types but also a new recombinant strain with a previously unpublished combination of adenovirus genomes. This strain was accepted by the Human Adenovirus Working Group as a novel genotype: HAdV-86. Seven strains that could not be typed with sufficient accuracy were also investigated using a PCR based on part of the fiber gene. By analysis of corresponding sequences of the 86 known HAdV genotypes, we determined that the proposed typing system should be able to distinguish all non-recombinant types, and with additional fiber information, all known HAdV genotypes.


Next-generation technologies and data analytical approaches for epigenomics.
Epigenetics refers to the collection of heritable features that modulate the genome-environment interaction without being encoded in the actual DNA sequence. While being mitotically and sometimes even meiotically transmitted, epigenetic traits often demonstrate extensive flexibility. This allows cells to acquire diverse gene expression patterns during differentiation, but also to adapt to a changing environment. However, epigenetic alterations are not always beneficial to the organism, as they are, for example, frequently identified in human diseases such as cancer. Accurate and cost-efficient genome-scale profiling of epigenetic features is thus of major importance to pinpoint these "epimutations," for example, to monitor the epigenetic impact of environmental exposure. Over the last decade, the field of epigenetics has been revolutionized by several innovative "epigenomics" technologies exactly addressing this need. In this review, we discuss and compare widely used next-generation methods to assess DNA methylation and hydroxymethylation, noncoding RNA expression, histone modifications, and nucleosome positioning. Although recent methods are typically based on "second-generation" sequencing, we also pay attention to still commonly used array- and PCR-based methods, and look forward to the additional advantages of single-molecule sequencing. As the current bottleneck in epigenomics research is the analysis rather than generation of data, the basic difficulties and problem-solving strategies regarding data preprocessing and statistical analysis are introduced for the different technologies. Finally, we also consider the complications associated with epigenomic studies of species with yet unsequenced genomes and possible solutions.


Hepatic cannabinoid receptor type 1 mediates alcohol-induced regulation of bile acid enzyme genes expression via CREBH.
Bile acids concentration in liver is tightly regulated to prevent cell damage. Previous studies have demonstrated that deregulation of bile acid homeostasis can lead to cholestatic liver disease. Recently, we have shown that ER-bound transcription factor Crebh is a downstream effector of hepatic Cb1r signaling pathway. In this study, we have investigated the effect of alcohol exposure on hepatic bile acid homeostasis and elucidated the mediatory roles of Cb1r and Crebh in this process. We found that alcohol exposure or Cb1r-agonist 2-AG treatment increases hepatic bile acid synthesis and serum ALT, AST levels in vivo alongwith significant increase in Crebh gene expression and activation. Alcohol exposure activated Cb1r, Crebh, and perturbed bile acid homeostasis. Overexpression of Crebh increased the expression of key bile acid synthesis enzyme genes via direct binding of Crebh to their promoters, whereas Cb1r knockout and Crebh-knockdown mice were protected against alcohol-induced perturbation of bile acid homeostasis. Interestingly, insulin treatment protected against Cb1r-mediated Crebh-induced disruption of bile acid homeostasis. Furthermore, Crebh expression and activation was found to be markedly increased in insulin resistance conditions and Crebh knockdown in diabetic mice model (db/db) significantly reversed alcohol-induced disruption of bile acid homeostasis. Overall, our study demonstrates a novel regulatory mechanism of hepatic bile acid metabolism by alcohol via Cb1r-mediated activation of Crebh, and suggests that targeting Crebh can be of therapeutic potential in ameliorating alcohol-induced perturbation of bile acid homeostasis.


The 2008 Canadian Hypertension Education Program recommendations for the management of hypertension: part 2 - therapy.
To update the evidence-based recommendations for the prevention and management of hypertension in adults. For lifestyle and pharmacological interventions, evidence was preferentially reviewed from randomized controlled trials and systematic reviews of trials. Changes in cardiovascular morbidity and mortality were the primary outcomes of interest. However, for lifestyle interventions, blood pressure lowering was accepted as a primary outcome given the lack of long-term morbidity and mortality data in this field. Progression of kidney dysfunction was also accepted as a clinically relevant primary outcome among patients with chronic kidney disease. A Cochrane collaboration librarian conducted an independent MEDLINE search from 2006 to August 2007 to update the 2007 recommendations. To identify additional published studies, reference lists were reviewed and experts were contacted. All relevant articles were reviewed and appraised independently by content and methodological experts using prespecified levels of evidence. For lifestyle modifications to prevent and treat hypertension, restrict dietary sodium intake to less than 100 mmol/day (and 65 mmol/day to 100 mmol/day in hypertensive patients); perform 30 min to 60 min of aerobic exercise four to seven days per week; maintain a healthy body weight (body mass index 18.5 kg/m(2) to 24.9 kg/m(2)) and waist circumference (smaller than 102 cm for men and smaller than 88 cm for women); limit alcohol consumption to no more than 14 units per week in men or nine units per week in women; follow a diet that is reduced in saturated fat and cholesterol, and one that emphasizes fruits, vegetables and low-fat dairy products, dietary and soluble fibre, whole grains and protein from plant sources; and consider stress management in selected individuals with hypertension. For the pharmacological management of hypertension, treatment thresholds and targets should be predicated on by the patient's global atherosclerotic risk, target organ damage and comorbid conditions. Blood pressure should be decreased to lower than 140/90 mmHg in all patients, and to lower than 130/80 mmHg in those with diabetes mellitus or chronic kidney disease. Most patients will require more than one agent to achieve these target blood pressures. For adults without compelling indications for other agents, initial therapy should include thiazide diuretics. Other agents appropriate for first-line therapy for diastolic and/or systolic hypertension include angiotensin-converting enzyme (ACE) inhibitors (in nonblack patients), long-acting calcium channel blockers (CCBs), angiotensin receptor antagonists (ARBs) or beta-blockers (in those younger than 60 years of age). A combination of two first-line agents may also be considered for initial treatment of hypertension if systolic blood pressure is 20 mmHg above target or if diastolic blood pressure is 10 mmHg above target. Other agents appropriate for first-line therapy for isolated systolic hypertension include long-acting dihydropyridine CCBs or ARBs. In patients with angina, recent myocardial infarction or heart failure, beta-blockers and ACE inhibitors are recommended as first-line therapy; in patients with cerebrovascular disease, an ACE inhibitor/diuretic combination is preferred; in patients with proteinuric nondiabetic chronic kidney disease, ACE inhibitors are recommended; and in patients with diabetes mellitus, ACE inhibitors or ARBs (or, in patients without albuminuria, thiazides or dihydropyridine CCBs) are appropriate first-line therapies. All hypertensive patients with dyslipidemia should be treated using the thresholds, targets and agents outlined in the Canadian Cardiovascular Society position statement (recommendations for the diagnosis and treatment of dyslipidemia and prevention of cardiovascular disease). Selected high-risk patients with hypertension but who do not achieve thresholds for statin therapy according to the position paper should nonetheless receive statin therapy. Once blood pressure is controlled, acetylsalicylic acid therapy should be considered. All recommendations were graded according to strength of the evidence and voted on by the 57 members of the Canadian Hypertension Education Program Evidence-Based Recommendations Task Force. All recommendations reported here achieved at least 95% consensus. These guidelines will continue to be updated annually.


A novel online didactic curriculum helps improve knowledge acquisition among non-emergency medicine rotating residents.
Rotating residents represent a significant proportion of housestaff in academic emergency departments (EDs), yet they rarely receive targeted didactic education during their emergency medicine (EM) rotations. The goals of this study were: 1) to determine the effectiveness of an online didactic curriculum in improving EM knowledge among rotating residents and 2) to assess rotating resident satisfaction with this curriculum. The authors created an online lecture series of six EM subject areas targeted to rotating residents called the Northwestern University Rotating Resident Curriculum (NURRC). All rotating residents at the study site were eligible, written consent was obtained, and the study was approved by the institutional review board. Consenting participants were pretested with a 42-question multiple-choice examination and then randomized to two groups: one with access to the NURRC during the first 2 weeks of the rotation (experimental) and one without (control). Halfway through the rotation, all participants were post-tested with a different multiple-choice examination, and the controls were then granted NURRC access. The primary outcome was the difference between pretest and posttest scores (score delta). The t-test was used to compare mean scores, and a linear regression model was used to determine the association of NURRC access on score delta after adjustment for pretest type and resident type. A postintervention survey was administered at the end of the rotation to assess satisfaction with the NURRC and collect suggestions for improvement. Fifty-four rotating residents were enrolled: 29 in the experimental group and 25 in the control group. There was no significant difference in pretest scores between the two groups. Mean score delta was 17.3% in the experimental group and 1.6% in the control group, an absolute difference of 15.7% (95% confidence interval [CI]=10% to 22%). After adjustment for resident type and pretest type, the only variable positively associated with the primary outcome was NURRC access. Third-year and preliminary-year internal medicine (IM) residents demonstrated the greatest absolute improvement in score delta when granted NURRC access. Eighty percent of the participants responded to the satisfaction survey. Over 80% of the survey respondents approved of each component lecture and of the NURRC overall. After exposure to an online didactic curriculum, rotating residents demonstrated a significant increase in EM knowledge and reported a high level of satisfaction with the didactic program.


Large right atrial myxoma containing gastric heterotopia presenting with dyspnea and bilateral leg edema due to pulmonary emboli and cardiovascular obstruction: the first known report of gastric heterotopia in the cardiovascular system.
A 52-year-old male presented with progressive dyspnea, bilateral leg edema, and elevated central venous pressure due to a large right atrial myxoma that caused vascular obstruction and pulmonary emboli. The myxoma contained gastric heterotopia. Other unusual features of this atrial myxoma included its large size, right atrial location, and attachment to the right atrial wall. Although gastric heterotopia has been reported throughout the gastrointestinal tract, and occasionally in other organs, this is the first report of gastric heterotopia in the cardiovascular system. This report confirms and extends previous reports of glandular elements or enteric glands within atrial, or cardiac, myxomas. The clinical presentation of the currently reported patient is explained as follows: the elevated central venous pressure resulted from cardiovascular obstruction and the dyspnea from multiple pulmonary emboli due to the large atrial myxoma. In this case, the clinical presentation was not attributable to the gastric heterotopia. The association of gastric heterotopia with atrial myxoma may, however, be clinically important because of the propensity of gastric heterotopia in the gastrointestinal tract to produce complications. The reported association may provide clues to the histogenesis of these two entities.


Identification of transcription factor genes involved in anthocyanin biosynthesis in carrot (Daucus carota L.) using RNA-Seq.
Anthocyanins are water-soluble colored flavonoids present in multiple organs of various plant species including flowers, fruits, leaves, stems and roots. DNA-binding R2R3-MYB transcription factors, basic helix-loop-helix (bHLH) transcription factors, and WD40 repeat proteins are known to form MYB-bHLH-WD repeat (MBW) complexes, which activates the transcription of structural genes in the anthocyanin pathway. Although black cultivars of carrots (Daucus carota L.) can accumulate large quantities of anthocyanin in their storage roots, the regulatory genes responsible for their biosynthesis are not well characterized. The current study aimed to analyze global transcription profiles based on RNA sequencing (RNA-Seq), and mine MYB, bHLH and WD40 genes that may function as positive or negative regulators in the carrot anthocyanin biosynthesis pathways. RNA was isolated from differently colored calli, as well as tissue samples from taproots of various black carrot cultivars across the course of development, and gene expression levels of colored and non-colored tissue and callus samples were compared. The expression of 32 MYB, bHLH and WD40 genes were significantly correlated with anthocyanin content in black carrot taproot. Of those, 11 genes were consistently up- or downregulated in a purple color-specific manner across various calli and cultivar comparisons. The expression of 10 out of these 11 genes was validated using real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). The results of this study provide insights into regulatory genes that may be responsible for carrot anthocyanin biosynthesis, and suggest that future focus on them may help improve our overall understanding of the anthocyanin synthesis pathway.


Trade-off between accuracy and interpretability for predictive in silico modeling.
Accuracy concerns the ability of a model to make correct predictions, while interpretability concerns to what degree the model allows for human understanding. Models exhibiting the former property are many times more complex and opaque, while interpretable models may lack the necessary accuracy. The trade-off between accuracy and interpretability for predictive in silico modeling is investigated. A number of state-of-the-art methods for generating accurate models are compared with state-of-the-art methods for generating transparent models. Results on 16 biopharmaceutical classification tasks demonstrate that, although the opaque methods generally obtain higher accuracies than the transparent ones, one often only has to pay a quite limited penalty in terms of predictive performance when choosing an interpretable model.


Neurotoxic and pharmacologic studies on enantiomers of the N-methylated analog of cathinone (methcathinone): a new drug of abuse.
These studies evaluated neurotoxic and pharmacologic properties of the R(+) and S(-) enantiomers of methcathinone, a psychostimulant drug that has surfaced in the illicit drug market, primarily in the S(-) form. Neurotoxic potential toward brain dopamine (DA) and serotonin (5-HT) neurons was assessed by measuring DA and 5-HT axonal markers and by means of silver degeneration studies; pharmacologic effects were evaluated by measuring locomotor stimulation. Methcathinone produced dose-related neurotoxic and locomotor stimulant effects which were species- and enantiomer-dependent. In mice, although both enantiomers produced toxic effects on DA neurons, the R(+) enantiomer was more potent, and neither enantiomer produced long-term effects on 5-HT neurons. By contrast, in behavioral studies, both enantiomers increased mouse locomotor activity, but the S(-) enantiomer was more potent, which suggests that methcathinone's neurotoxic and locomotor stimulant effects may be separable. Additional studies were done with rats, because mice are often refractory to 5-HT neurotoxicity induced by amphetamines. In the rat, both enantiomers produced toxic effects on DA neurons, only S(-)-methcathinone produced toxic effects on 5-HT neurons, and both enantiomers produced comparable locomotor stimulant effects. Together, these results indicate that: 1) Methcathinone has the potential to damage DA and 5-HT neurons; 2) Methcathinone neurotoxicity is enantiomer and species dependent; 3) Methcathinone's neurotoxic and locomotor stimulant effects are dissociable in mice but not rats; and 4) N-methylation confers 5-HT toxic activity onto cathinone, the N-desmethyl derivative of methcathinone, which is known to lack 5-HT neurotoxic activity.


Trans-acting translational regulatory RNA binding proteins.
The canonical molecular machinery required for global mRNA translation and its control has been well defined, with distinct sets of proteins involved in the processes of translation initiation, elongation and termination. Additionally, noncanonical, trans-acting regulatory RNA-binding proteins (RBPs) are necessary to provide mRNA-specific translation, and these interact with 5' and 3' untranslated regions and coding regions of mRNA to regulate ribosome recruitment and transit. Recently it has also been demonstrated that trans-acting ribosomal proteins direct the translation of specific mRNAs. Importantly, it has been shown that subsets of RBPs often work in concert, forming distinct regulatory complexes upon different cellular perturbation, creating an RBP combinatorial code, which through the translation of specific subsets of mRNAs, dictate cell fate. With the development of new methodologies, a plethora of novel RNA binding proteins have recently been identified, although the function of many of these proteins within mRNA translation is unknown. In this review we will discuss these methodologies and their shortcomings when applied to the study of translation, which need to be addressed to enable a better understanding of trans-acting translational regulatory proteins. Moreover, we discuss the protein domains that are responsible for RNA binding as well as the RNA motifs to which they bind, and the role of trans-acting ribosomal proteins in directing the translation of specific mRNAs. This article is categorized under: RNA Interactions with Proteins and Other Molecules > RNA-Protein Complexes Translation > Translation Regulation Translation > Translation Mechanisms.


A20, ABIN-1/2, and CARD11 mutations and their prognostic value in gastrointestinal diffuse large B-cell lymphoma.
Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous group of aggressive lymphomas with the activated B-cell-like subtype characterized by constitutive NF-κB activation. Activating mutations of CARD11 and inactivating mutations of A20 are frequent events in DLBCL. However, the full extent of genetic alterations in the NF-κB pathway regulators and their potential prognostic value in DLBCL remain to be investigated. We investigated the genetic abnormalities of CARD11, A20, and ABIN-1/2/3 (the A20 binding inhibitor of NF-κB) and their clinicopathologic correlation in gastrointestinal DLBCL. The somatic mutation and copy number changes of CARD11, A20, and ABIN-1/2/3 were investigated in 71 gastrointestinal DLBCLs by PCR/sequencing, and interphase FISH/array comparative genomic hybridization, respectively. The mutations identified were functionally characterized by NF-κB reporter assays and immunoprecipitation experiments. Recurrent somatic mutations were found in CARD11 (10%), A20 (17%), ABIN-1 (4%), and ABIN-2 (3%), but not in ABIN-3. In comparison with the wild-type, all CARD11 mutants were potent NF-κB activators in vitro. On the basis of the destructive nature of the observed mutations, and the findings by reporter assays and immunoprecipitation studies, most if not all of the somatic mutations that were seen in A20, ABIN-1, and ABIN-2 could impair their normal functions. Among these genetic abnormalities, A20 somatic mutation was significantly associated with both poor overall survival and event-free survival. We show further evidence of NF-κB pathway genetic abnormalities in DLBCL, which are potentially valuable in the prognosis and design of future therapeutic strategies.


Granger causal time-dependent source connectivity in the somatosensory network.
Exploration of transient Granger causal interactions in neural sources of electrophysiological activities provides deeper insights into brain information processing mechanisms. However, the underlying neural patterns are confounded by time-dependent dynamics, non-stationarity and observational noise contamination. Here we investigate transient Granger causal interactions using source time-series of somatosensory evoked magnetoencephalographic (MEG) elicited by air puff stimulation of right index finger and recorded using 306-channel MEG from 21 healthy subjects. A new time-varying connectivity approach, combining renormalised partial directed coherence with state space modelling, is employed to estimate fast changing information flow among the sources. Source analysis confirmed that somatosensory evoked MEG was mainly generated from the contralateral primary somatosensory cortex (SI) and bilateral secondary somatosensory cortices (SII). Transient Granger causality shows a serial processing of somatosensory information, 1) from contralateral SI to contralateral SII, 2) from contralateral SI to ipsilateral SII, 3) from contralateral SII to contralateral SI, and 4) from contralateral SII to ipsilateral SII. These results are consistent with established anatomical connectivity between somatosensory regions and previous source modeling results, thereby providing empirical validation of the time-varying connectivity analysis. We argue that the suggested approach provides novel information regarding transient cortical dynamic connectivity, which previous approaches could not assess.


One-frame two-dimensional deflectometry for phase retrieval by addition of orthogonal fringe patterns.
Deflectometry is a well-known method to characterize pure phase objects by measuring the deformation of fringes. In principle, the retrieved magnitude is the partial derivative of the phase along the coordinate orthogonal to the fringes. In order to recover the phase it is necessary to know the derivatives in two orthogonal directions, which is usually achieved by rotating 90° the original fringes and acquiring a new deformed pattern. This "time-multiplexed" two-dimensional deflectometry is a time-consuming operation if the goal is to characterize phase objects in real time. In the present paper we propose a kind of two-dimensional deflectometry that allows acquisition of fringe patterns in two orthogonal directions in a single frame. The proposed procedure utilizes a two-dimensional ("additive") fringe pattern that allows the application of Takeda's method to each coordinate independently. The advantage of the method (with respect to the traditional one) is that it simplifies the setup and reduces the acquisition time. Validation experiments are presented.


Rapid start-up and efficient long-term nitritation of low strength ammonium wastewater with a sequencing batch reactor containing immobilized cells.
Major concerns about nitritation of low-strength ammonium wastewaters include low ammonium loading rates (ALRs) (usually below 0.2 kg/m³-d) and uncertainty with the long-term stability of the process. The purpose of this study was to test a sequencing batch reactor filled with cell-immobilized polyethylene glycol (PEG) pellets (∼2 mm in size) to see if it could achieve efficient and stable nitritation under various environmental conditions. The sequencing batch reactor (SBR) was fed with synthetic ammonium wastewater of 30 ± 2 mg-N/L and pH 8 ± 0.05, maintaining the dissolved oxygen (DO) concentration at 1.7 ± 0.2 mg/L and the temperature at 30 ± 1 °C. The reaction was easily converted to partial nitrification mode within a month by feeding a relatively high ammonium substrate (∼ 100 mg-N/L) in the beginning. We observed stable nitritation over 300 days with high ALRs (as high as ∼1.1 kg-N/m³-d), nitrite accumulation rates (mostly over 97%), and ammonium removal rates (mostly over 95%). DO was the major limiting substrate when the DO concentration was below ∼4 mg/L and the NH4⁺-N concentration was above ∼ 5 mg/L, giving an almost linear increase in the ammonium oxidation rate with the bulk DO increase. Low temperatures mainly affected the reaction rate, which could be compensated for by increasing the pellet volume (i.e. biomass). Our results demonstrated that an SBR filled with small cell-immobilized PEG pellets could achieve very efficient and stable nitritation of a low-strength ammonium wastewater.


The risk of subjective symptoms in mobile phone users in Poland--an epidemiological study.
To assess the type and incidence of subjective symptoms related to the use of mobile phones in Polish users. The study was conducted in 2005 using a questionnaire survey. Although it has been quite a long time, up to now, no such data have been published for Poland. The questionnaire consisted of 53 questions concerning sex, age, education, general health, characteristics of a mobile phone (hand-held, loud-speaking unit) as well as the habits associated with its use (frequency and duration of calls, text messages, etc.) and complaints associated with using a mobile phone. As many as 1800 questionnaires were sent. The response was obtained from 587 subjects aged 32.6 ± 11.3 (48.9% women, 51.1% men); the age did not differ significantly between men and women. The subjects owned a cell phone for an average of 3 years. Majority of the respondents used the phone intensively, i.e. daily (74%) or almost daily (20%). Headaches were reported significantly more often by the people who talked frequently and long in comparison with other users (63.2% of the subjects, p = 0.0029), just like the symptoms of fatigue (45%, p = 0.013). Also, the feeling of warmth around the ear and directly to the auricle was reported significantly more frequently by the intensive mobile phone users, compared with other mobile phone users (47.3%, p = 0.00004 vs. 44.6%, p = 0.00063, respectively). Most symptoms appeared during or immediately after a call and disappeared within 2 h after the call. Continuous headache, persisting for longer than 6 h since the end of a call, was reported by 26% of the subjects. Our results show that the mobile phone users may experience subjective symptoms, the intensity of which depends on the intensity of use of mobile phones.


Lifestylisation of the social: The government of diabetes care in Denmark.
Since the 1970s, the public authorities of many OECD countries have emphasised the need for preventing lifestyle diseases and promoting the vigour of their populations. Based on the Foucauldian analytics of dispositive, we critically address some of the normative implications of the preventive interventions in the area of type 2 diabetes care. Through an analysis of public health documents from 1981 to 2016, it is shown that the government of lifestyle was extended and institutionalised by a reform of the Danish public sector in 2007. Following the reform, rationalities of public health policies sought to prevent unhealthy lifestyles not only through individual behaviour but also through the social surroundings of citizens. In contrast to the claim that we are seeing a retraction of state responsibility and interventions in the area of public health, it is suggested that we are witnessing an expansion in state ambitions expressed through a lifestyle dispositive. These ambitions are less about transferring the responsibility to the individual and more about governing and mobilising the social relations and environments of type 2 diabetes patients and citizens in general to make the everyday choice of a healthy lifestyle easier.


A simulation study for the analysis of uncertain binary responses: application to first insemination success in beef cattle.
A simulation was carried out to investigate the methods of analyzing uncertain binary responses for success or failure at first insemination. A linear mixed model that included, herd, year, and month of mating as fixed effects; and unrelated service sire, sire and residual as random effects was used to generate binary data. Binary responses were assigned using the difference between days to calving and average gestation length. Females deviating from average gestation length lead to uncertain binary responses. Thus, the methods investigated were the following: (1) a threshold model fitted to certain (no uncertainty) binary data (M1); (2) a threshold model fitted to uncertain binary data ignoring uncertainty (M2); and (3) analysis of uncertain binary data, accounting for uncertainty from day 16 to 26 (M3) or from day 14 to 28 (M4) after introduction of the bull, using a threshold model with fuzzy logic classification. There was virtually no difference between point estimates obtained from M1, M3, and M4 with true values. When uncertain binary data were analyzed ignoring uncertainty (M2), sire variance and heritability were underestimated by 22 and 24%, respectively. Thus, for noisy binary data, a threshold model contemplating uncertainty is needed to avoid bias when estimating genetic parameters.


Rfc5, a small subunit of replication factor C complex, couples DNA replication and mitosis in budding yeast.
The inhibition of DNA synthesis prevents mitotic entry through the action of the S phase checkpoint. In the yeast Saccharomyces cerevisiae, an essential protein kinase, Spk1/Mec2/Rad53/Sad1, controls the coupling of S phase to mitosis. In an attempt to identify genes that genetically interact with Spk1, we have isolated a temperature-sensitive mutation, rfc5-1, that can be suppressed by overexpression of SPK1. The RFC5 gene encodes a small subunit of replication factor C complex. At the restrictive temperature, rfc5-1 mutant cells entered mitosis with unevenly separated or fragmented chromosomes, resulting in loss of viability. Thus, the rfc5 mutation defective for DNA replication is also impaired in the S phase checkpoint. Overexpression of POL30, which encodes the proliferating cell nuclear antigen, suppressed the replication defect of the rfc5 mutant but not its checkpoint defect. Taken together, these results suggested that replication factor C has a direct role in sensing the state of DNA replication and transmitting the signal to the checkpoint machinery.


Self-cleaning properties in engineered sensors for dopamine electroanalytical detection.
Fouling and passivation are the major drawbacks for a wide applicability of electroanalytical sensors based on nanomaterials, especially in biomedical and environmental fields. The production of highly engineered devices, designed ad hoc for specific applications, is the key factor in the direction of overcoming the problem and accessing effective sensors. Here, the fine-tuning of the system, composed of a highly ordered distribution of silver nanoparticles between a bottom silica and a top titania layer, confers multifunctional properties to the device for a biomedical complex challenge: dopamine detection. The crucial importance of each component towards a robust and efficient electroanalytical system is studied. The total recovery of the electrode performance after a simple UV-A cleaning step (self-cleaning), due to the photoactive interface and the aging resistance, is deeply investigated.


Structure and expression of a complementary DNA for the nuclear coded precursor of human mitochondrial ornithine transcarbamylase.
Most mitochondrial proteins are encoded in the nucleus and are translated on free cytoplasmic ribosomes as larger precursors containing amino-terminal "leader" sequences, which are removed after the precursors are taken up by mitochondria. We have deduced the complete primary structure of the precursor of a human mitochondrial matrix enzyme, ornithine transcarbamylase (OTC), from the nucleotide sequence of cloned complementary DNA. The amino-terminal leader peptide of OTC is 32 amino acids in length and contains four arginines but no acidic residues. Cleavage of the leader peptide from the "mature" protein occurs between glutamine and asparagine residues. The sequence of mature human OTC resembles that of the subunits of both OTC and aspartate transcarbamylase from Escherichia coli. The biological activity of the cloned OTC complementary DNA was tested by joining it with SV40 (an animal virus) regulatory elements and transfecting cultured HeLa cells, which do not normally express OTC. Both the precursor and mature forms of the OTC subunit were identified; in stable transformants, enzymatic activity was also detected.


Integration of metabolomics and in vitro metabolism assays for investigating the stereoselective transformation of triadimefon in rainbow trout.
Triadimefon is a systemic agricultural fungicide of the triazole class whose major metabolite, triadimenol, also a commercial fungicide, provides the majority of the actual fungicidal activity, i.e., inhibition of steroid demethylation. Both chemicals are chiral: triadimefon has one chiral center with two enantiomers while its enzymatic reduction to triadimenol produces a second chiral center and two diastereomers with two enantiomers each. All six stereoisomers of the two fungicides were separated from each other using a chiral BGB-172 column on a GC-MS system so as to follow stereospecificity in metabolism by rainbow trout hepatic microsomes. In these microsomes the S-(+) enantiomer of triadimefon was transformed to triadimenol 27% faster than the R-(-) enantiomer, forming the four triadimenol stereoisomers at rates different from each other. The most fungi-toxic stereoisomer (1S,2R) was produced at the slowest rate; it was detectable after 8 h, but below the level of method quantitation. The triadimenol stereoisomer ratio pattern produced by the trout microsomes was very different from that of the commercial triadimenol standard, in which the most rat-toxic pair of enantiomers (known as "Diastereomer A") is about 85% of the total stereoisomer composition. The trout microsomes produced only about 4% of "Diastereomer A". Complementary metabolomic studies with NMR showed that exposure of the separate triadimefon enantiomers and the racemate to rainbow trout for 48 h resulted in different metabolic profiles in the trout liver extracts, i.e., different endogenous metabolite patterns that indicated differences in effects of the two enantiomers.


Family history of cancer and cancer risks in women with BRCA1 or BRCA2 mutations.
Women who carry a deleterious mutation in BRCA1 or BRCA2 have high lifetime risks of breast and ovarian cancers. However, the influence of a family history of these cancers on these risks in women with BRCA mutations is unclear. We calculated cancer incidence rates for a multinational cohort comprising 3011 women with BRCA1 or BRCA2 mutations who were followed up for a mean of 3.9 years, during which time 243 incident breast or ovarian cancers were recorded. The 10-year cumulative risks of breast cancer were 18.1% (95% confidence interval [CI] = 13.3% to 22.8%) for women with a BRCA1 mutation and 15.2% (95% CI = 9.1% to 21.2%) for women with a BRCA2 mutation. Among women with a BRCA1 mutation, the risk of breast cancer increased by 1.2-fold for each first-degree relative with breast cancer before age 50 years (hazard ratio [HR] = 1.21; 95% confidence interval [CI] = 0.94 to 1.57) and the risk of ovarian cancer increased by 1.6 fold for each first- or second-degree relative with ovarian cancer (HR = 1.61; 95% CI = 1.21 to 2.14). Among women with a BRCA2 mutation, the risk of breast cancer increased by 1.7-fold for each first-degree relative younger than 50 years with breast cancer (HR = 1.67; 95% CI = 1.04 to 2.07).


Reduction in breast cancer mortality from organized service screening with mammography: 1. Further confirmation with extended data.
In an earlier publication, our evaluation of data from breast cancer screening programs in seven Swedish counties suggested a 40% reduction in incidence-based breast cancer mortality among women actually screened. In the current study, we expand the previous analysis from seven counties to 13 large areas within nine counties, including six of the original counties and seven additional areas, examine a longer period of follow-up (20-44 years), apply new analytic methods for the evaluation of incidence-based breast cancer mortality, and estimate the number needed to screen to save one life. Data from six of the original counties (one being excluded as it does not yet have 10 years of follow-up after the initiation of screening), with increased follow-up, and seven additional large areas, within three counties, representing approximately 45% of Swedish women, provide information about age at diagnosis, age at death, and screening history for 542,187 women in the prescreening and 566,423 women in the screening epochs. Regardless of year of diagnosis, there were a total of 6,231 deaths due to breast cancer in the period of study as a whole. Of these, 4,778 were incidence-based deaths in the two epochs, i.e., death among cases diagnosed within either the prescreening or screening period. Data were analyzed using Poisson regression and adjusted, when necessary, for self-selection bias, contemporaneous changes in incidence, and changes in mortality independent of screening. Attendance was uniformly high, averaging 75% in the screening epochs. Recall rates for assessment varied from 4% to 5% at the first round of screening and approximately 3% at later rounds. Detection rates averaged five breast cancers per 1,000 women screened in the first round, and four breast cancers per 1,000 women screened in subsequent rounds. There was a significant 45% reduction in incidence-based breast cancer mortality among screened women in the screening epoch relative to incidence-based breast cancer mortality in the prescreening epoch (relative risk, 0.55; 95% confidence intervals, 0.51-0.59). After adjusting for self-selection bias, there still was a significant 43% reduction in incidence-based breast cancer mortality associated with screening (relative risk, 0.57; 95% confidence intervals, 0.53-0.62). These results indicate a reduction in breast cancer mortality of between 40% and 45% in association with screening, after adjustment for self-selection bias. These results were obtained with modest human costs: the number needed to screen to save one life was estimated as 472.


Improved specificity of ICA assays in the Fourth International Immunology of Diabetes Serum Exchange Workshop.
The goal of the Fourth International Workshop for Standardization of ICA Measurements was to determine the specificity of ICA assays and their ability to distinguish between control sera (n = 57) and sera from IDDM-related individuals--representing relatives of IDDM patients (n = 21), healthy individuals who later developed IDDM (n = 8), or newly diagnosed IDDM patients (n = 23). Results from 28 laboratories were analyzed. The mean specificity (percentage of control sera reported as negative) among 27 laboratories was 91%, including 6 laboratories with 100% specificity. Nevertheless, 78% of laboratories found at least one control sample > 0 JDF U. Among samples from first-degree relatives, the mean concordance was 86%, including three sera found negative (0 JDF U) by all laboratories. Among individuals who later developed diabetes, the mean concordance was 93%, with two sera found positive by 100% of laboratories. In sera from newly diagnosed IDDM patients, the mean concordance was 82%. Three sera were found positive and one serum negative by all laboratories. The JDF U of the sera considered to be positive were significantly greater than each laboratory's average for the controls. In conclusion, the results from laboratories participating in the Fourth International ICA Workshop demonstrated excellent specificity, good concordance, and an ability to separate control sera from defined, IDDM-related subjects.