Influence of regulatory measures on the rate of spontaneous adverse drug reaction reporting in Italy.
The reporting of adverse drug reactions (ADRs) is the mainstay of post-marketing surveillance systems. Under-reporting and selective reporting are considered the main limitations of a spontaneous reporting-based pharmacovigilance system. However, excessive reporting induced by external events may also impair signal detection by increasing the noise level. The aim of this study was to examine the influence of regulatory measures and other external factors on the rate of ADR reporting in Italy, focusing on four situations occurring in the last 10 years: ACE inhibitor-induced cough; HMG-CoA reductase inhibitors ('statins') and rhabdomyolysis; nimesulide and hepatic toxicity; and cyclo-oxygenase (COX)-2 selective inhibitors ('coxibs') and increase in cardiovascular risk. The study was based on data from spontaneous reporting in six Italian regions collected from January 1995 to December 2005. We analysed a 10-year period as a reasonable time interval around the four situations of interest, highlighting the influence of regulatory measures on the rate of ADR reporting (number of reports per million inhabitants). Chi-squared tests were used to assess the statistical significance of any changes in ADR reporting. Drug sales data were also studied to examine possible changes in drug use. Sales data were expressed as daily defined dose per 1000 inhabitants per day. ACE inhibitors: a 5-fold increase in the reporting rate of ACE inhibitor-induced cough was observed in 1998 and 1999 following a restriction on reimbursement for angiotensin receptor blockers introduced in 1998 and removed at the end of 1999. Statins: after the withdrawal of cerivastatin in 2001, the ADR reporting rate increased more than 4-fold, with musculoskeletal ADRs representing about 60% of all the ADRs reported in that year, and progressively decreased in the following years. Nimesulide: an increase in hepatic ADR reporting was observed after withdrawal of the drug from the Finnish and Spanish markets in 2002. Coxibs: no important changes in the rate of cardiovascular events reporting in the period 2000-4 were observed. In 2005, after the withdrawal of rofecoxib in September 2004, both the ADR reporting rate and sales of the drug decreased drastically. Our data suggest that spontaneous ADR reporting can be influenced in different ways by external events. Our data emphasize the need for educational initiatives aimed at increasing the doctor's and patient's awareness of the usefulness and the limitations of spontaneous reporting in the pharmacovigilance system. Such initiatives should use appropriate risk communication strategies in order to avoid unnecessary alarm, which could cause unjustified interruption of therapies or misplaced confidence in new drugs.


[Intracellular accumulation and antitumor activity of various nanosomal formulations of paclitaxel].
Paclitaxel-loaded poly(lactic-glycolic) copolymer nanoparticles have been prepared using a precipitation technique. The cytotoxic activity of nanosomal paclitaxel was studied on the model of highly resistant cell line Jurkat WT (human T-cell leukemia) using various biochemical assays. It is found that the inhibitory concentration (IC50) for the experimental formulation of paclitaxel falls within 10(-4)-10(-6) M. Accumulation of nanoparticles in the highly resistant Jurkat/WT cells was revealed by fluorescence microscopy.


Bone Strength of the Calcaneus Is Associated with Dietary Calcium Intake in Older Japanese Men, but Not Women.
The relationship between calcium intake and bone strength in older Asian individuals, including Japanese, is controversial; therefore, we herein investigated this relationship in older Japanese populations. We performed a cross-sectional analysis of 314 participants older than 65 years who voluntarily participated in a medical examination and responded to questionnaires. The osteo-sono assessment index (OSI) measured at the right calcaneus using a quantitative ultrasonic device was used as an indicator of bone strength. The daily dietary intake of calcium was assessed using a brief-type self-administered diet history questionnaire. A two-way analysis of covariance revealed a significant interaction between sex and calcium intake on the OSI (p < 0.01). A multiple regression analysis showed a positive correlation between calcium intake and the OSI in males (p < 0.01), but not females (p = 0.27). In females, grip strength divided by body weight positively correlated with the OSI (p = 0.04). The present results suggest that a higher calcium intake contributes to bone strength in older Japanese males. Although a higher grip strength may contribute to bone strength in females, the potential of estrogen as a confounding factor needs to be considered.


Towards a global cancer knowledge network: dissecting the current international cancer genomic sequencing landscape.
While next generation sequencing has enhanced our understanding of the biological basis of malignancy, current knowledge on global practices for sequencing cancer samples is limited. To address this deficiency, we developed a survey to provide a snapshot of current sequencing activities globally, identify barriers to data sharing and use this information to develop sustainable solutions for the cancer research community. A multi-item survey was conducted assessing demographics, clinical data collection, genomic platforms, privacy/ethics concerns, funding sources and data sharing barriers for sequencing initiatives globally. Additionally, respondents were asked as to provide the primary intent of their initiative (clinical diagnostic, research or combination). Of 107 initiatives invited to participate, 59 responded (response rate = 55%). Whole exome sequencing (P = 0.03) and whole genome sequencing (P = 0.01) were utilized less frequently in clinical diagnostic than in research initiatives. Procedures to identify cancer-specific variants were heterogeneous, with bioinformatics pipelines employing different mutation calling/variant annotation algorithms. Measurement of treatment efficacy varied amongst initiatives, with time on treatment (57%) and RECIST (53%) being the most common; however, other parameters were also employed. Whilst 72% of initiatives indicated data sharing, its scope varied, with a number of restrictions in place (e.g. transfer of raw data). The largest perceived barriers to data harmonization were the lack of financial support (P < 0.01) and bioinformatics concerns (e.g. lack of interoperability) (P = 0.02). Capturing clinical data was more likely to be perceived as a barrier to data sharing by larger initiatives than by smaller initiatives (P = 0.01). These results identify the main barriers, as perceived by the cancer sequencing community, to effective sharing of cancer genomic and clinical data. They highlight the need for greater harmonization of technical, ethical and data capture processes in cancer sample sequencing worldwide, in order to support effective and responsible data sharing for the benefit of patients.


Trends in sinusitis research: a systematic review of extramural funding.
Innovation represents a core value of the American Rhinologic Society (ARS), with multiple efforts to promote research in the advancement rhinologic care. We therefore sought to identify trends in extramural sinusitis funding and underutilized sources of support to facilitate future efforts. A systematic review of the National Institutes of Health (NIH) Research Portfolio Online Tools (RePORTER) database (fiscal year 1993 to 2017) was completed with the search strategy: ("chronic sinusitis" OR rhinosinusitis). All identified studies were accepted for review, with comparison to ARS membership rolls to identify studies supported by ARS investigators. Foundation awards were surveyed to identify and characterize additional sources of support. The systematic review identified 958 projects receiving NIH funding, of which 120 remain active. The percentage of sinusitis-related awards and total funding relative to all NIH awards increased over the past 10 years (2006 to 2016) from 0.06% (8 / 9128) and 0.09% ($2,151,152 / $3,358,338,602) to 0.87% (86 / 9540) and 0.90% ($37,201,095 / $4,300,145,614). Among active studies, 9 investigators maintain membership in the ARS and serve as principal investigator or project leader in 12 (10%) studies. ARS investigators received the greatest number of awards from the National Institute on Deafness and Other Communication Disrders (n = 8,66.7%), while only receiving 2.2% of awarded funding from the National Institute of Allergy and Infectious Diseases ($607,500/$26,873,022), the largest source of awards for sinusitis research. Support for sinusitis research is significantly growing, with the largest source of active funding not being fully utilized by members of the ARS. Further efforts to promote funding priorities among extramural sources is necessary to facilitate increased funding for ARS member initiatives.


Local increases in mechanical tension shape compartment boundaries by biasing cell intercalations.
Mechanical forces play important roles during tissue organization in developing animals. Many tissues are organized into adjacent, nonmixing groups of cells termed compartments. Boundaries between compartments display a straight morphology and are associated with signaling centers that are important for tissue growth and patterning. Local increases in mechanical tension at cell junctions along compartment boundaries have recently been shown to prevent cell mixing and to maintain straight boundaries. The cellular mechanisms by which local increases in mechanical tension prevent cell mixing at compartment boundaries, however, remain poorly understood. Here, we have used live imaging and quantitative image analysis to determine cellular dynamics at and near the anteroposterior compartment boundaries of the Drosophila pupal abdominal epidermis. We show that cell mixing within compartments involves multiple cell intercalations. Frequency and orientation of cell intercalations are unchanged along the compartment boundaries; rather, an asymmetry in the shrinkage of junctions during intercalation is biased, resulting in cell rearrangements that suppress cell mixing. Simulations of tissue growth show that local increases in mechanical tension can account for this bias in junctional shrinkage. We conclude that local increases in mechanical tension maintain cell populations separate by influencing junctional rearrangements during cell intercalation.


Morally Problematic Situations Encountered by Adults Living With Rare Diseases.
Rare diseases are generally poorly understood from scientific and medical standpoints due, to their complexity and low prevalence. As a result, individuals living with rare diseases struggle to obtain timely diagnoses and suitable care. These clinical difficulties add to the physical and psychological impacts of living with chronic and often severe medical conditions. From the standpoint of pragmatist ethics, the morally problematic situations that adults living with rare diseases experience matter crucially. However, there is little known about these experiences. A survey study was conducted with 121 adults living with rare diseases in Québec, Canada, to identify morally problematic situations encountered in the healthcare system and everyday life as part of a participatory action research project. Morally problematic situations elicited internal tensions and constraints to agency. Adults living with rare diseases experienced morally problematic situations of stigmatization, disbelief, and sometimes abuse in the healthcare system. These situations were compounded by diagnostic delays, inadequate care, and suboptimal follow-up, and led some individuals to opt-out of medical care. In their personal lives, these individuals sometimes found themselves in situations of physical and financial dependency. They often also had to give up professional occupations, academic training, or life projects. Adults living with rare diseases experience important morally problematic situations navigating the healthcare system and their everyday lives, some of which could be alleviated through interventions developed through future participatory action research.


Protection of Fecal Microbiota Transplantation in a Mouse Model of Multiple Sclerosis.
Given the growing evidence of a link between gut microbiota (GM) dysbiosis and multiple sclerosis (MS), fecal microbiota transplantation (FMT), aimed at rebuilding GM, has been proposed as a new therapeutic approach to MS treatment. To evaluate the viability of FMT for MS treatment and its impact on MS pathology, we tested FMT in mice with experimental autoimmune encephalomyelitis (EAE), a mouse model of MS. We provide evidence that FMT can rectify altered GM to some extent with a therapeutic effect on EAE. We also found that FMT led to reduced activation of microglia and astrocytes and conferred protection on the blood-brain barrier (BBB), myelin, and axons in EAE. Taken together, our data suggest that FMT, as a GM-based therapy, has the potential to be an effective treatment for MS.


Active kallikrein response to changes in sodium-chloride intake in essential hypertensive patients.
To evaluate the behavior of active kallikrein excretion in salt-sensitive and salt-resistant hypertensive patients during changes in sodium-chloride (NaCl) intake, 61 male, nonobese, nondiabetic outpatients affected by uncomplicated essential hypertension were given a diet that contained 140 mmol NaCl per day for 2 wk. Patients then received either a low- (20 mmol NaCl/day) or a high- (320 mmol NaCl/day) sodium diet for 2 wk, according to a randomized, double-blind, cross-over protocol. Hypertensive patients were classified as salt sensitive when their diastolic blood pressure rose by at least 10 mm Hg after the high-sodium diet, and decreased by at least 10 mm Hg after the low-sodium diet, considering as baseline blood pressure values those that were taken at the end of the 140 mmol NaCl/day intake period. The remaining patients were classified as salt resistant or, when diastolic blood pressure increased by 10 mm Hg or more after low-sodium intake, as counter-regulating. Twenty-three patients were therefore classified as salt sensitive, 28 as salt resistant, and 10 as counter-regulating. The baseline active kallikrein excretion was significantly lower (P < 0.0001) in salt-sensitive (0.62 +/- 0.31 U/24 h) patients than in salt-resistant (1.39 +/- 0.44 U/24 h) and counter-regulating patients (1.27 +/- 0.38 U/24 h). Surprisingly, the kallikrein response to changes in sodium intake was similar in all subgroups, although enzyme excretion was always at the lowest level in salt-sensitive hypertensive patients. This latter group also showed the highest plasma atrial natriuretic peptide levels (28.2 +/- 8.5 fmol/mL, P < 0.0001 versus salt-resistant and counter-regulating patients), and the greatest peptide increment with sodium load (P < 0.0001 versus salt-resistant and counter-regulating patients). Counter-regulating patients showed the steepest increase in plasma renin activity (from 0.24 +/- 0.18 to 0.83 +/- 0.21 ng/L per s, P < 0.001) and decrease of plasma atrial natriuretic peptide (from 26.1 +/- 6.3 to 6.8 +/- 3.1 fmol/mL, P < 0.001) when switched from a high to a low-sodium intake. In conclusion, salt-sensitive hypertensive patients excrete less active kallikrein than do salt-resistant and counter-regulating patients, but maintain a normal enzyme response to changes in dietary sodium intake. The exaggerated response of atrial natriuretic peptide to high-sodium intake that was observed in the same patients could be compensating for an impaired renal capability to excrete a sodium load.


Effect of sodium benzoate on polymorphonuclear leukocyte function.
Sodium benzoate is widely used as a food preservative and reputed to be a scavenger of hydroxyl radical (.OH). The effects of sodium benzoate on the function of polymorphonuclear leukocytes (PMN) stimulated by S. aureus or the chemical agent, phorbol myristate acetate (PMA) were examined in vitro using assays of chemiluminescence (CL), total bactericidal activity, intracellular recovery of bacteria, as well as release of lactate dehydrogenase (LDH), lysozyme and superoxide anion (O2(-)). Sodium benzoate decreased chemiluminescence, superoxide anion and lysozyme release by PMN stimulated with S. aureus but did not similarly affect these responses of PMN to the chemical agent, PMA. The ability of PMN to kill S. aureus was also impaired by sodium benzoate and associated with a reduced number of intracellular bacteria recovered after 90 minutes incubation. LDH release from PMN was not demonstrable at concentrations of sodium benzoate below 100 mM indicating that damage can not account for these findings. The underlying mechanism of the altered bactericidal function of PMN treated with sodium benzoate appears to be a result of decreased uptake of S. aureus.


Benchmarking B cell epitope prediction: underperformance of existing methods.
Sequence profiling is used routinely to predict the location of B-cell epitopes. In the postgenomic era, the need for reliable epitope prediction is clear. We assessed 484 amino acid propensity scales in combination with ranges of plotting parameters to examine exhaustively the correlation of peaks and epitope location within 50 proteins mapped for polyclonal responses. After examining more than 10(6) combinations, we found that even the best set of scales and parameters performed only marginally better than random. Our results confirm the null hypothesis: Single-scale amino acid propensity profiles cannot be used to predict epitope location reliably. The implication for studies using such methods is obvious.


Isolation and characterization of the mating-type idiomorphs from the wheat septoria leaf blotch fungus Mycosphaerella graminicola.
Both mating-type loci from the wheat septoria leaf blotch pathogen Mycosphaerella graminicola have been cloned and sequenced. The MAT1-2 gene was identified by screening a genomic library from the MAT1-2 isolate IPO94269 with a heterologous probe from Tapesia yallundae. The MAT1-2 idiomorph is 2772 bp and contains a single gene encoding a putative high-mobility-group protein of 394 amino acids. The opposite idiomorph was obtained from isolate IPO323, which has the complementary mating type, by long-range PCR using primers derived from sequences flanking the MAT1-2 idiomorph. The MAT1-1 locus is 2839 bp in size and contains a single open reading frame encoding a putative alpha1-domain protein of 297 amino acids. Within the nonidiomorphic sequences, homology was found with palI, encoding a membrane receptor from Aspergillus nidulans, and a gene encoding a putative component of the anaphase-promoting complex from Schizosaccharomyces pombe and a DNA-(apurinic or apyrimidinic) lyase from S. pombe. For each of the MAT genes specific primers were designed and tested on an F1 mapping population that was generated from a cross between IPO323 and IPO94269. An absolute correlation was found between the amplified allele-specific fragments and the mating type as determined by backcrosses of each F1 progeny isolate to the parental isolates. The primers were also used to screen a collection of field isolates in a multiplex PCR. An equal distribution of MAT1-1 and MAT1-2 alleles was found for most geographic origins examined.


Differential effect of botulinal toxin on esophageal motor function in infants.
The effect of Clostridium botulinum toxin on esophageal motor function was studied in four infants (ages 5-9 months) with confirmed infant botulism. Esophageal motility studies using a perfused catheter assembly were performed during the acute phase in all patients, and during the recovery phase in one patient. Motor function of the proximal esophagus and upper esophageal sphincter was abnormal in each, while motor function of the distal esophagus and lower esophageal sphincter was normal. Mean lower esophageal sphincter pressure was 24 mm Hg for the group (normal: 15-30 mm Hg). Sequential studies of proximal esophageal motility in one infant revealed a return of normal motor function which correlated with recovery of peripheral muscle strength and gag reflexes. C. botulinum toxin impairs proximal, but not distal, esophageal motor function in the infant botulism model. This effect appears to be consistent with the known action of the toxin on synaptic acetylcholine release and current concepts regarding distribution of cholinergic and noncholinergic neurotransmitter receptors in the esophagus.


Informal Caregiving Relationships in Psychosis: Reviewing the Impact of Patient Violence on Caregivers.
A modest association can be found between people with a schizophrenia spectrum diagnosis (psychosis) and perpetrating acts of violence. When a person with psychosis does engage in violence, it is their informal carers, when compared to those from the general population, who are more likely to be the targets, and violence will often occur within the family home. Despite the importance of carer support for improving patient outcomes, our understanding of how carers are impacted by patient initiated violence in psychosis remains limited. This paper reviews literature documenting the effects of patient-initiated violence in psychosis on carer functioning. The review comprised searches of Medline, PsychInfo, Embase, and Web of Science databases and the hand searches of reference lists from relevant published papers. The review was limited to English language publications from inception to 11th September 2017, and where carer experiences following reports of violence from patients with psychosis were specifically recorded. Data from 20 papers using mixed methodologies were reviewed. Patient violence in psychosis was linked to poorer carer outcomes, including carer reports of burden, trauma, fear, and helplessness. There is, however, a significant need for further studies to systematically quantify the impact and correlates of patient initiated violence on psychosis caregivers, and improve prevention.


Effects of probiotic supplementation over 5 months on routine haematology and clinical chemistry measures in healthy active adults.
Use of probiotic-containing foods and probiotic supplements is increasing; however, few studies document safety and tolerability in conjunction with defined clinical end points. This paper reports the effects of 150 days of supplementation with either a single- (Bifidobacterium animalis subsp. lactis Bl-04) or a double-strain (Lactobacillus acidophilus NCFM and Bifidobacterium animalis subsp. lactis Bi-07) probiotic on routine haematology and clinical chemistry measures in healthy active adults. Pre- to post-intervention changes in laboratory measures were determined and compared between supplement and placebo groups. Overall there were few differences in routine haematology and clinical chemistry measures between supplement and placebo groups post-intervention. Exceptions included plasma calcium (P=0.03) and urea (P=0.015); however, observed changes were small and within assay-specific laboratory reference ranges. These data provide evidence supporting the use of these probiotic supplements over a period of 5 months in healthy active adults without obvious safety or tolerability issues.


Applying a research ethics committee approach to a medical practice controversy: the case of the selective COX-2 inhibitor rofecoxib.
The new class of anti-inflammatory drugs, the COX-2 inhibitors, have been commercially successful to the point of market dominance within a short time of their launch. They attract a price premium on the basis that they are associated with fewer adverse gastric events than traditional anti-inflammatory drugs. This marketing continues even though a pivotal safety study with one of the COX-2 inhibitors, rofecoxib, showed a significant increase in myocardial infarction with rofecoxib use compared with a traditional anti-inflammatory drug. This finding has led to a series of publications containing pooled analyses of existing data that both support and refute the possibility of increased cardiovascular risk with COX-2 inhibitors. These medical journal publications have served to obfuscate rather than provide guidance for medical practitioners. Consideration of a research ethics committee approach to this issue suggests that it would deal with the controversy in a straightforward manner-namely, it would simply inform research participants of the trial results with rofecoxib. The certainty of this research ethics committee approach raises the issue of whether it should be applied in normal medical practice outside of the research environment. A consideration of the legal tests for disclosure of information suggests that therapeutic medical practice should mirror that within the research environment, in this case.


Passively transferred myasthenia gravis: protection of mouse endplates by Fab fragments from human myasthenic IgG.
Using the mouse passive transfer model, the effects of purified human myasthenic immunoglobulin G and of the monovalent Fab fragment on neuromuscular junctions were investigated. Treatment with IgG markedly reduced amplitudes of miniature endplate potentials. When Fab fragments were transferred alone or with subsequent addition of IgG no neuromuscular transmission block was induced. Myasthentic IgG and Fab were specifically demonstrated at the neuromuscular junctions by immunohistochemistry. On electronmicroscopy endplate structure was normal in transfer experiments using IgG for up to 30 days. It is suggested that Fab fragments bind to acetylcholine receptors without affecting transmission and protect them from the attack of complete IgG antibodies.


Presence of intestinal intraepithelial lymphocytes in mice with severe combined immunodeficiency disease.
The murine intestinal epithelium contains a heterogeneous population of intraepithelial leukocytes (IEL) most of which are granulated, Thy-1-CD5-CD8+. In order to assess the lineage relationship of this subgroup of IEL to peripheral T cells, we examined IEL in mice with the severe combined immunodeficiency (scid/scid) mutation, which lack T and B cells in peripheral lymphoid tissues. Electron and light microscopy showed that the intestine from scid/scid mice had granulated IEL similar to IEL in normal C.B-17 mice. Flow cytometry of isolated IEL stained with monoclonal antibodies against Thy-1, CD3, CD4, CD5 and CD8 showed that scid/scid mice IEL contained cells with the Thy-1-CD4-CD5-CD8+ phenotype. Immunohistochemical staining of IEL in tissue sections with antibodies to Thy-1 and CD8 confirmed that the Thy-1-CD8+ cells were in the intestinal epithelium. These scid/scid IEL also lacked CD3 expression and mRNA for the V gamma 7 V region gene of the gamma T cell receptor. We conclude that scid/scid mice contain precursors for IEL that can differentiate into a granulated Thy-1-CD5-CD8+ IEL in the intestine. The absence of CD8+ peripheral T cells in these mice suggests that these IEL differ from classical T cells in their ability to differentiate and express CD8 and do not require T cell receptor expression for their localization to the intestine.


Adherence to Report and Patient Perception of an Interactive App for Managing Symptoms During Radiotherapy for Prostate Cancer: Descriptive Study of Logged and Interview Data.
Patients undergoing radiotherapy for prostate cancer experience symptoms related to both the cancer itself and its treatment, and it is evident that patients with prostate cancer have unmet supportive care needs related to their disease. Over the past decade, there has been an increase in the amount of research within the field of mobile health and the use of apps as tools for managing illness. The main challenge is to develop a mobile technology to its full potential of being interactive in real time. The interactive app Interaktor, which aims to identify and manage symptoms in real time includes (1) a function for patients' assessment of the occurrence, frequency, and distress of symptoms; (2) a connection to a monitoring Web interface; (3) a risk assessment model that sends alerts via text message to health care providers; (4) continuous access to evidence-based self-care advice and links to relevant websites for more information; and (5) graphs for the patients and health care providers to view the history of symptom reporting. The aim of the study was to investigate user behavior, adherence to reporting, and the patients' experiences of using Interaktor during radiotherapy for localized advanced prostate cancer. The patients were instructed to report daily during the time of treatment and then for an additional 3 weeks. Logged data from patients' use of the app were analyzed with descriptive statistics. Interview data about experiences of using the app were analyzed with content analysis. A total of 66 patients participated in the study. Logged data showed that adherence to daily reporting of symptoms was high (87%). The patients used all the symptoms included in the app. Of the reports, 15.6% generated alerts to the health care providers. Overall, the patients found that it was easy and not particularly time-consuming to send a daily report, and many described it as becoming a routine. Reporting symptoms facilitated reflection on their symptoms and gave them a sense of security. Few technological problems were reported. The use of Interaktor increased patients' sense of security and their reflections on their own well-being and thereby served as a supportive tool for the self-management of symptoms during treatment of prostate cancer. Some further development of the app's content might be beneficial for future use.


ATR-p53 restricts homologous recombination in response to replicative stress but does not limit DNA interstrand crosslink repair in lung cancer cells.
Homologous recombination (HR) is required for the restart of collapsed DNA replication forks and error-free repair of DNA double-strand breaks (DSB). However, unscheduled or hyperactive HR may lead to genomic instability and promote cancer development. The cellular factors that restrict HR processes in mammalian cells are only beginning to be elucidated. The tumor suppressor p53 has been implicated in the suppression of HR though it has remained unclear why p53, as the guardian of the genome, would impair an error-free repair process. Here, we show for the first time that p53 downregulates foci formation of the RAD51 recombinase in response to replicative stress in H1299 lung cancer cells in a manner that is independent of its role as a transcription factor. We find that this downregulation of HR is not only completely dependent on the binding site of p53 with replication protein A but also the ATR/ATM serine 15 phosphorylation site. Genetic analysis suggests that ATR but not ATM kinase modulates p53's function in HR. The suppression of HR by p53 can be bypassed under experimental conditions that cause DSB either directly or indirectly, in line with p53's role as a guardian of the genome. As a result, transactivation-inactive p53 does not compromise the resistance of H1299 cells to the interstrand crosslinking agent mitomycin C. Altogether, our data support a model in which p53 plays an anti-recombinogenic role in the ATR-dependent mammalian replication checkpoint but does not impair a cell's ability to use HR for the removal of DSB induced by cytotoxic agents.


Chromosome abnormalities associated with salivary gland epithelial cell lines transformed in vitro and in vivo with evidence of a role for genetic imbalance in transformation.
Chromosomal abnormalities associated with five in vitro-transformed male mouse salivary gland epithelial cell lines were compared with those in three cell lines derived from in vivo-induced tumors. All cell lines were hypotetraploid. Structural chromosome abnormalities were found in all cell lines, but no consistent aberration was detected. Nevertheless, losses of chromosomes 1, 4, 7, 9, and 14 were observed in all of the in vitro-transformed cell lines. With the possible exception of chromosome 1, the same chromosome losses were noted in the in vivo-transformed cell lines. In addition, a consistent feature of both in vitro- and in vivo-transformed cell lines was the presence of double minute chromosomes and homogeneously staining regions. Where both of these chromosome types were present in the same cell line, they were mutually exclusive. The Y chromosome was absent in nearly all of the cell lines. These findings are consistent with the view that, in salivary gland epithelium, the malignant phenotype may result from a genetic imbalance caused by specific chromosome losses from tetraploid cells.


First isolation of Leptospira fainei serovar Hurstbridge from two human patients with Weil's syndrome.
Leptospira fainei serovar Hurstbridge is a recently discovered Leptospira species and so far it has only been cultured from animal sources. Based on positive serology and positive PCR for L. fainei among patients suspected of having leptospirosis, a role in human disease seems likely. This study describes two patients with Weil's disease from whom L. fainei was cultured. A local source of the infections was suspected, as these two patients resided in the same area of Denmark, were hospitalised approximately at the same time and had not been travelling recently. The Leptospira species was determined by serology, PCR and sequencing of bacterial DNA. One patient developed autoimmune hepatitis in the course of the L. fainei infection and was treated with both antibiotics and immunosuppression with good effect. The other patient had a self-limiting disease and did not receive any treatment.


Theorizing the Role of Dopaminergic Polymorphic Risk Alleles with Intermittent Explosive Disorder (IED), Violent/Aggressive Behavior and Addiction: Justification of Genetic Addiction Risk Severity (GARS) Testing.
Scientific studies have provided evidence that there is a relationship between violent and aggressive behaviors and addictions. Genes involved with the reward system, specifically the brain reward cascade (BRC), appear to be associated with various addictions and impulsive, aggressive, and violent behaviors. In our previous research, we examined the Taq A1 allele (variant D2 dopamine receptor gene) and the DAT-40 base repeat (a variant of the dopamine transporter gene) in 11 Caucasian boys at the Brown School in San Marcus, Texas, diagnosed with intermittent explosive disorder. Thirty supernormal controls were screened to exclude several reward-deficit behaviors, including pathological violence, and genotyped for the DRD2 gene. Additionally, 91 controls were screened to exclude ADHD, pathological violence, alcoholism, drug dependence, and tobacco abuse, and their results were compared with DAT1 genotype results. In the schoolboys vs. supercontrols, there was a significant association with the D2 variant and a trend with the dopamine transporter variant. Results support our hypothesis and the involvement of at least two gene risk alleles with adolescent violent/aggressive behaviors. This study and the research presented in this paper suggest that violent/aggressive behaviors are associated with a greater risk of addiction, mediated via various genes linked to the BRC. This review provides a contributory analysis of how gene polymorphisms, especially those related to the brain reward circuitry, are associated with violent behaviors.


Effect of a hospital policy of not accepting free infant formula on in-hospital formula supplementation rates and breast-feeding duration.
To investigate the effect of public hospitals in Hong Kong not accepting free infant formula from manufacturers on in-hospital formula supplementation rates and breast-feeding duration. Prospective cohort study. In-patient postnatal units of four public hospitals in Hong Kong. Two cohorts of breast-feeding mother-infant pairs (n 2560). Cohort 1 (n 1320) was recruited before implementation of the policy to stop accepting free infant formula and cohort 2 (n 1240) was recruited after policy implementation. Participants were followed prospectively for 12 months or until they stopped breast-feeding. The mean number of formula supplements given to infants in the first 24 h was 2·70 (sd 3·11) in cohort 1 and 1·17 (sd 1·94) in cohort 2 (P<0·001). The proportion of infants who were exclusively breast-fed during the hospital stay increased from 17·7 % in cohort 1 to 41·3 % in cohort 2 (P<0·001) and the risk of breast-feeding cessation was significantly lower in cohort 2 (hazard ratio=0·81; 95 % CI 0·73, 0·90). Participants who non-exclusively breast-fed during the hospital stay had a significantly higher risk of stopping any or exclusive breast-feeding. Higher levels of formula supplementation also increased the risk of breast-feeding cessation in a dose-response pattern. After implementation of a hospital policy to pay market price for infant formula, rates of in-hospital formula supplementation were reduced and the rates of in-hospital exclusive breast-feeding and breast-feeding duration increased.


Toxic effects of T-2 toxin on reproductive system in male mice.
The present study was conducted to evaluate the effects of T-2 toxin on semen quality, fertility and serum testosterone concentration in mice. Adult male mice were mated with sexually mature untreated female mice after being exposed to intraperitoneal injection of T-2 toxin at 0, 5, 10 or 15 mg/kg body weight daily for 7 successive days. Semen quality, serum testosterone concentration and fertility of treated mice were assessed. The results showed that the number of abnormal spermatozoa increased significantly and a significant decrease in spermatozoa with integrated acrosome was observed in males treated with T-2 toxin at all doses, As well, the amount of live spermatozoa decreased significantly in mice treated with 10 and 15 mg/kg body weight T-2 toxin. Low pregnancy rate and high fetal resorption rate were observed when females were mated with T-2 toxin-exposed males. Testicular and cauda epididymal sperm counts, efficiency of sperm production and serum testosterone concentration were significantly reduced in mice treated with T-2 toxin at all doses in a dose-dependent manner. In conclusion, these findings indicated that T-2 toxin presented toxic effects on reproductive system of adult male mice.


Selected acculturation factors and birth defects in the National Birth Defects Prevention Study, 1997-2011.
Acculturation has been examined with respect to various pregnancy adverse outcomes, including birth defects. Given the mixed and limited findings on the association between nativity and birth defects, we sought to further explore parental nativity and years lived in the U.S. across a range of defects. Data from the National Birth Defects Prevention Study were used for this analysis. Infants with one of 46 major isolated birth defects (30 noncardiac/16 cardiac conditions) and infants without birth defects (controls) born during 1997-2011 were included. We examined parental nativity (foreign-born mothers, fathers, and both parents combined compared to a referent of both U.S.-born parents) and the number of years lived in the U.S. (≤5/6+ years). Descriptive statistics and logistic regression analyses were performed to estimate crude/adjusted odds ratios and 95% confidence intervals. Compared to U.S.-born mothers, foreign-born mothers tended to be older (25+ years), of Hispanic or Other race/ethnicity and were less likely to have reported drinking, smoking, illicit drug use, or having taken folic acid. In the adjusted analysis, seven findings among both parents reporting a foreign-birth were significant, including an increased association with spina bifida, anotia/microtia, and diaphragmatic hernia (aORs range: 1.3-1.7), and a reduced association with craniosynostosis and gastroschisis (aORs = 0.7). A generally protective effect was observed among foreign-born subjects living in the U.S. ≤5 years. We found that nativity was associated with some selected isolated defects, although the direction of effect varied by phenotype and by a number of years residing in the U.S.


Phosphorylation of Glutamine Synthetase on Threonine 301 Contributes to Its Inactivation During Epilepsy.
The astrocyte-specific enzyme glutamine synthetase (GS), which catalyzes the amidation of glutamate to glutamine, plays an essential role in supporting neurotransmission and in limiting NH4 + toxicity. Accordingly, deficits in GS activity contribute to epilepsy and neurodegeneration. Despite its central role in brain physiology, the mechanisms that regulate GS activity are poorly defined. Here, we demonstrate that GS is directly phosphorylated on threonine residue 301 (T301) within the enzyme's active site by cAMP-dependent protein kinase (PKA). Phosphorylation of T301 leads to a dramatic decrease in glutamine synthesis. Enhanced T301 phosphorylation was evident in a mouse model of epilepsy, which may contribute to the decreased GS activity seen during this trauma. Thus, our results highlight a novel molecular mechanism that determines GS activity under both normal and pathological conditions.


Essential trace metals and heavy metals in newly diagnosed schizophrenic patients and those on anti-psychotic medication.
Nutritional deprivation in the early stage of life increases the risk of developing schizophrenia. Oxidative stress, disturbed thinking and irrational behavior which are common to schizophrenic patients may be a result of changes in the levels of certain trace metals. Twenty (20) healthy volunteers and a total of thirty-five (35) schizophrenic patients consisting of 20 on antipsychotic drugs for at least 2 weeks and 15 newly diagnosed but not taking antipsychotic drugs were considered. The plasma levels of trace metals were analyzed using atomic absorption spectrophotometer. Fe and Se were significantly reduced in newly diagnosed and medicated-schizophrenic patients compared with controls. Pb, Cd and Cr were significantly raised in newly diagnosed drug free schizophrenic patients compared with controls. While Cr and Cd were significantly raised in schizophrenic patients on treatment compared with the controls. Levels of certain nutritionally essential trace metals (Fe and Se) were reduced while levels of certain heavy metals (Pb, Cr and Cd) were raised in schizophrenic patients.


Capillary refill time is an unreliable indicator of cardiovascular status in term neonates.
Decisions regarding the need for volume replacement in neonates often are made in the immediate newborn period. Capillary refill time (CRT) is used as an indicator of circulatory status; however, recent data show that CRT varies considerably with age, ambient and skin temperature, anatomical site of measurement, and duration of pressure. The purpose of this study was to (1) examine the relationship between CRT and heart rate (HR) and blood pressure (BP) in term neonates, and (2) evaluate the differences among CRT values measured at 3 body sites and with varying duration of cutaneous pressure. This was a prospective, cross-sectional, correlational study. Subjects Forty-two appropriate-weight-for-gestational-age (AGA) neonates with birthweights, (M = 3407; SD = +/- 540 g), gestational ages (M = 39 weeks; SD = +/- 1 week), and sex (21 males, 21 females). Infants had no history of perinatal distress or maternal chorioamnionitis. Each neonate was studied prospectively 1 to 4 hours after birth. The infants were clothed with only a diaper and evaluated on a radiant warmer bed set to achieve an axillary temperature of 36.5 degrees to 37.0 degrees C. Capillary refill time was measured with a digital stopwatch at 3 sites: volar surface of finger (F), plantar surface of heel (H), and lower sternum (St), using brief (1- to 2-second) and extended (3- to 4-second) pressure. Heart rate was auscultated and counted for 60 seconds, and BP was measured by oscillometry. Relationships among variables were assessed by Pearson correlation coefficient, analysis of variance, and multiple regression analysis. The Bonferroni correction for multiple comparisons was applied. Capillary refill time, blood pressure, and heart rate. There was no significant site variation for CRT for either brief (2.4 +/- 0.6 to 2.9 +/- 1.0 seconds) or extended (3.8 +/- 0.8 to 4.3 +/- 0.8 seconds) pressure. However, regardless of site, CRT was greater when extended versus brief pressure was used (P < 0.001). There were no significant correlations between HR and CRT. There was a moderate, direct relationship between BP and CRT observed in the following anatomic sites: (1) sternum/extended pressure and systolic BP (SBP), diastolic BP, and mean BP (r = 0.35, P = 0.02; r = 0.49, P = 0.001; and r = 0.43, P = 0.005, respectively); (2) sternum/brief pressure and SBP (r = 0.31, P = 0.05); and (3) finger/extended pressure and SBP (r = 0.30, P = 0.05). An unanticipated moderate, direct correlation between BP and CRT was observed; prolongation of CRT occurred with elevated blood pressure. This finding may have been secondary to increased circulating vasoactive substances in the newborn period; measurement of these substances was beyond the scope of this study. In addition, CRT was highly dependent on the duration of cutaneous pressure, regardless of the site. These 2 findings indicate that CRT may be an unreliable indicator of cardiovascular status in the term neonate during the first 4 hours after birth.


A PKM2 signature in the failing heart.
A salient feature of the failing heart is metabolic remodeling towards predominant glucose metabolism and activation of the fetal gene program. Sunitinib is a multitargeted receptor tyrosine kinase inhibitor used for the treatment of highly vascularized tumors. In diabetic patients, sunitinib significantly decreases blood glucose. However, a considerable proportion of sunitinib-treated patients develop cardiac dysfunction or failure. We asked whether sunitinib treatment results in shift towards glycolysis in the heart. Glucose uptake by the heart was increased fivefold in mice treated with sunitinib. Transcript analysis by qPCR revealed an induction of genes associated with glycolysis and reactivation of the fetal gene program. Additionally, we observed a shift in the enzyme pyruvate kinase from the adult M1 (PKM1) isoform to the fetal M2 (PKM2) isoform, a hallmark of the Warburg Effect. This novel observation led us to examine whether a similar shift occurs in human heart failure. Examination of tissue from patients with heart failure similarly displayed an induction of PKM2. Moreover, this phenomenon was partially reversed following mechanical unloading. We propose that pyruvate kinase isoform switching represents a novel feature of the fetal gene program in the failing heart.


Antimicrobial resistance among lower respiratory tract isolates of Haemophilus influenzae: results of a 1992-93 western Europe and USA collaborative surveillance study. The Alexander Project Collaborative Group.
During 1992 and 1993, 2718 respiratory tract isolates of Haemophilus influenzae were obtained from two study centres in each of five West European countries and five study centres in the USA. beta-Lactamase production was assessed and MICs of 14 antimicrobial agents determined in a single co-ordinating laboratory using a broth microdilution method in Mueller-Hinton-lysed horse blood medium. The prevalence of strains producing beta-lactamase varied between 0 and 37.9%. In general, the highest prevalence was in study centres from Spain and the USA with slightly lower rates observed in France and the UK. Only a single confirmed beta-lactamase-negative, ampicillin resistant strain was recovered during the entire study. Erythromycin resistance, defined as an MIC of > or = 4.0 mg/L, was noted in 57.5% of isolates. Among the other antimicrobials tested, resistance rates > or = 1.0% were observed only with cefaclor (3.7%), chloramphenicol (1.4%) and co-trimoxazole (2.5%). In no case, was the prevalence of resistance or beta-lactamase production significantly greater in 1993 than in 1992.


A novel subviral agent associated with a geminivirus: the first report of a DNA satellite.
Numerous plant RNA viruses have associated with them satellite (sat) RNAs that have little or no nucleotide sequence similarity to either the viral or host genomes but are completely dependent on the helper virus for replication. We report here on the discovery of a 682-nt circular DNA satellite associated with tomato leaf curl geminivirus (TLCV) infection in northern Australia. This is the first demonstration that satellite molecules are not limited to RNA viral systems. The DNA satellite (TLCV sat-DNA) is strictly dependent for replication on the helper virus replication-associated protein and is encapsidated by TLCV coat protein. It has no significant open reading frames, and it shows no significant sequence similarity to the 2766-nt helper-virus genome except for two short motifs present in separate putative stem-loop structures: TAATATTAC, which is universally conserved in all geminiviruses, and AATCGGTGTC, which is identical to a putative replication-associated protein binding motif in TLCV. Replication of TLCV sat-DNA is also supported by other taxonomically distinct geminiviruses, including tomato yellow leaf curl virus, African cassava mosaic virus, and beet curly top virus. Therefore, this unique DNA satellite does not appear to strictly conform with the requirements that dictate the specificity of interaction of geminiviral replication-associated proteins with their cognate origins as predicted by the current model of geminivirus replication.


Under-use of emergency contraception for victims of sexual assault.
Approximately 700,000 women in the reproductive age group are victims of sexual assault in the United States per year. Between 1% and 5% of sexual assaults result in pregnancy, for a total of 32,000 pregnancies per year. Of these, 14,000 are aborted because of incest or rape. To determine the percent of emergency departments in the state of Pennsylvania offering routine counseling and provision of emergency contraception to victims of sexual assault. Secondary objectives were to compare provision practices for Catholic versus non-Catholic hospitals, and to compare these practices with other services, such as sexually transmitted disease prophylaxis and sexual assault counseling. A 15-item survey instrument was designed to determine the volume of sexual assault patients seen per year, routinely offered services, and emergency contraception protocols. Three telephone callers administered surveys, using a pre-designed script for each call. Of the 165 eligible hospitals, 125 (76%) replied. Less than half (42%) of all hospitals routinely offer emergency contraception counseling, and 16% of the hospitals did not offer any counseling regarding emergency contraception. Provision of emergency contraception to victims of sexual assault is inconsistent and insufficient. It is important that sexual assault patients not be further victimized by a system that fails to meet their needs.


Workplace bullying and mental distress - a prospective study of Norwegian employees.
Using a prospective design, the objective of this study was to determine the relationship between workplace bullying and mental distress. Altogether, 1971 Norwegian employees, recruited from 20 organizations, answered questions regarding workplace bullying and mental distress at both baseline and follow-up. Baseline data were gathered between 2004-2006, and follow-up data were gathered between 2006-2009. The time-lag between baseline and follow-up was approximately two years for all the respondents in all the organizations. The factors measured in the study were individual characteristics, mental distress measured with the Hopkins Symptom Checklist (HSCL-10), self-reported workplace bullying measured with a single item from the General Nordic Questionnaire for Psychological and Social Factors at Work (QPSNordic) and job demands and job control assessed by QPSNordic. A multiple linear regression analysis adjusted for mental distress, sex, age, job demands and job control at baseline [β=0.05, 95% confidence interval (95% CI) 0.03-0.17] and a repeated measures ANOVA adjusted for sex and age [F(3,1965)=38.37; partial η (2)=0.06] showed that workplace bullying predicted mental distress. Furthermore, a multiple binary logistic regression analysis adjusted for bullying, sex, age, job demands and job control at baseline [odds ratio (OR) 2.30, 95% CI 1.43-3.69] showed that mental distress was a predictor of bullying. We found support for the notion that self-reported workplace bullying is a predictor of mental distress two years later. Bullying had an independent effect on mental distress after adjusting for job demands and job control. Mental distress was also found to be a predictor of bullying, indicating that the reverse relationship is also important.


Pressure-Regulated Gene Expression and Enzymatic Activity of the Two Periplasmic Nitrate Reductases in the Deep-Sea Bacterium Shewanella piezotolerans WP3.
Shewanella species are widely distributed in marine environments, from the shallow coasts to the deepest sea bottom. Most Shewanella species possess two isoforms of periplasmic nitrate reductases (NAP-α and NAP-β) and are able to generate energy through nitrate reduction. However, the contributions of the two NAP systems to bacterial deep-sea adaptation remain unclear. In this study, we found that the deep-sea denitrifier Shewanella piezotolerans WP3 was capable of performing nitrate respiration under high hydrostatic pressure (HHP) conditions. In the wild-type strain, NAP-β played a dominant role and was induced by both the substrate and an elevated pressure, whereas NAP-α was constitutively expressed at a relatively lower level. Genetic studies showed that each NAP system alone was sufficient to fully sustain nitrate-dependent growth and that both NAP systems exhibited substrate and pressure inducible expression patterns when the other set was absent. Biochemical assays further demonstrated that NAP-α had a higher tolerance to elevated pressure. Collectively, we report for the first time the distinct properties and contributions of the two NAP systems to nitrate reduction under different pressure conditions. The results will shed light on the mechanisms of bacterial HHP adaptation and nitrogen cycling in the deep-sea environment.


Gastrointestinal protective effect of dietary spices during ethanol-induced oxidant stress in experimental rats.
Spices are traditionally known to have digestive stimulant action and to cure digestive disorders. In this study, the protective effect of dietary spices with respect to activities of antioxidant enzymes in gastric and intestinal mucosa was examined. Groups of Wistar rats were fed for 8 weeks with diets containing black pepper (0.5%), piperine (0.02%), red pepper (3.0%), capsaicin (0.01%), and ginger (0.05%). All these spices significantly enhanced the activities of antioxidant enzymes--superoxide dismutase, catalase, glutathione reductase, and glutathione-S-transferase--in both gastric and intestinal mucosa, suggesting a gastrointestinal protective role for these spices. In a separate study, these dietary spices were found to alleviate the diminished activities of antioxidant enzymes in gastric and intestinal mucosa under conditions of ethanol-induced oxidative stress. The gastroprotective effect of the spices was also reflected in their positive effect on mucosal glycoproteins, thereby lowering mucosal injury. The amelioration of the ethanol-induced decrease in the activities of antioxidant enzymes in gastric and intestinal mucosa by dietary spices suggests their beneficial gastrointestinal protective role. This is the first report on the gastrointestinal protective potential of dietary spices.


DC-SIGN interactions with human immunodeficiency virus type 1 and 2 and simian immunodeficiency virus.
Dendritic cells (DCs) efficiently bind and transmit human immunodeficiency virus (HIV) to cocultured T cells and so may play an important role in HIV transmission. DC-SIGN, a novel C-type lectin that is expressed in DCs, has recently been shown to bind R5 HIV type 1 (HIV-1) strains and a laboratory-adapted X4 strain. To characterize the interaction of DC-SIGN with primate lentiviruses, we investigated the structural determinants of DC-SIGN required for virus binding and transmission to permissive cells. We constructed a panel of DC-SIGN mutants and established conditions which allowed comparable cell surface expression of all mutants. We found that R5, X4, and R5X4 HIV-1 isolates as well as simian immunodeficiency and HIV-2 strains bound to DC-SIGN and could be transmitted to CD4/coreceptor-positive cell types. DC-SIGN contains a single N-linked carbohydrate chain that is important for efficient cell surface expression but is not required for DC-SIGN-mediated virus binding and transmission. In contrast, C-terminal deletions removing either the lectin binding domain or the repeat region abrogated DC-SIGN function. Trypsin-EDTA treatment inhibited DC-SIGN mediated infection, indicating that virus was maintained at the surface of the DC-SIGN-expressing cells used in this study. Finally, quantitative fluorescence-activated cell sorting analysis of AU1-tagged DC-SIGN revealed that the efficiency of virus transmission was strongly affected by variations in DC-SIGN expression levels. Thus, variations in DC-SIGN expression levels on DCs could greatly affect the susceptibility of human individuals to HIV infection.


2019 John Charnley Award: Increased risk of prosthetic joint infection following primary total knee and hip arthroplasty with the use of alternative antibiotics to cefazolin: the value of allergy testing for antibiotic prophylaxis.
The aims of this study were to characterize antibiotic choices for perioperative total knee arthroplasty (TKA) and total hip arthroplasty (THA) prophylaxis, assess antibiotic allergy testing efficacy, and determine rates of prosthetic joint infection (PJI) based on perioperative antibiotic regimen. We evaluated all patients undergoing primary TKA or THA at a single academic institution between January 2004 and May 2017, yielding 29 695 arthroplasties (22 705 patients), with 3411 arthroplasties in 2576 patients (11.5%) having undergone preoperative allergy testing. A series of institutional databases were combined to identify allergy consultation outcomes, perioperative antibiotic regimen, and infection-free survivorship until final follow-up. Among 2576 allergy-tested patients, 2493 patients (97%) were cleared to use cephalosporins. For the entire cohort, 28 174 arthroplasties (94.9%) received cefazolin and 1521 (5.1%) received non-cefazolin antibiotics. Infection-free survivorship was significantly higher among arthroplasties receiving cefazolin compared with non-cefazolin antibiotics, with 0.06% higher survival free of infection at one month, 0.56% at two months, 0.61% at one year, and 1.19% at ten years (p < 0.001). Overall, the risk of PJI was 32% lower in patients treated with cefazolin after adjusting for the American Society of Anesthesiologists (ASA) classification, joint arthroplasty (TKA or THA), and body mass index (BMI; p < 0.001). The number needed to treat with cefazolin to prevent one PJI was 164 patients at one year and 84 patients at ten years. Therefore, potentially 6098 PJIs could be prevented by one year and 11 905 by ten years in a cohort of 1 000 000 primary TKA and THA patients. PJI rates are significantly higher when non-cefazolin antibiotics are used for perioperative TKA and THA prophylaxis, highlighting the positive impact of preoperative antibiotic allergy testing to increase cefazolin usage. Given the low rate of true penicillin allergy positivity, and the readily modifiable risk factor that antibiotic choice provides, we recommend perioperative testing and clearance for all patients presenting with penicillin and cephalosporin allergies. Cite this article: Bone Joint J 2019;101-B(6 Supple B):9-15.


Effect of placement technique on the push-out bond strength of calcium-silicate based cements.
The purpose of this study was to evaluate the effects of ultrasonic and manual placement techniques on the push-out bond strength of Biodentine and MTA with and without calcium chloride. One hundred and twenty mid-root slices from forty freshly extracted single-rooted human mandibular premolar teeth were instrumented and randomly divided into six groups (n=20) according to the filling material and placement technique applied, as follows: G1: MTA-manual compaction, G2: Biodentine-manual compaction, G3: MTA+5% CaCl2-manual compaction, G4: MTA-ultrasonic activation, G5: Biodentine-ultrasonic activation, G6: MTA+5% CaCl2-ultrasonic activation. The push-out bond strengths were measured using an Instron testing machine. Data were analyzed using twoway analysis of variance (ANOVA) with Bonferroni correction. The ultrasonic activation significantly enhanced the bond strength values of the materials. Biodentine presented higher bond strength values than that of MTA groups. The addition of CaCl2 to MTA did not improve the bond strength of the material.


Backtracking of colloids: a multiparticle collision dynamics simulation study.
The role of sound in the dynamics of mesoscale systems is typically neglected, since frequently the associated time scales are much smaller than all the other time scales of interest. However, for sufficiently small objects embedded in a solvent with a sufficiently small sound velocity, sound can play a crucial role. In particular, behavior resembling viscoelasticity has been theoretically predicted for nonviscoelastic fluids. This effect is due to the interference of the propagation of sound waves caused by the solute particle's motion and hydrodynamic vortex formation. We demonstrate this effect, known as backtracking, in computer simulations employing the method of multiparticle collision dynamics. We systematically study the influence of sound on the dynamics of the solute particle, and find that it disappears in the long-time limit. Thus, we confirm that sonic effects at the single-particle level can be neglected at sufficiently long times.


Functional characterization and subcellular localization of the 16K cysteine-rich suppressor of gene silencing protein of tobacco rattle virus.
The 16 kDa cysteine-rich protein (16K) of tobacco rattle virus (TRV) is known to partially suppress RNA silencing in Drosophila cells. In this study, we show that 16K suppresses RNA silencing in green fluorescent protein (GFP)-transgenic Nicotiana benthamiana plants using an Agrobacterium-mediated transient assay. 16K slightly reduced the accumulation of short interfering RNAs (siRNA) of GFP, suggesting that the protein may interfere with the initiation and/or maintenance of RNA silencing. Deletion of either the N- or C-terminal part of 16K indicated that the entire 16K open reading frame (ORF) is necessary for its silencing suppression function. Pentapeptide insertion scanning mutagenesis (PSM) revealed that only two short regions of 16K tolerated five extra amino acid insertions without considerable reduction in its silencing suppression function. The tolerant regions coincide with sequence variability between tobravirus cysteine-rich proteins, indicating a strong functional and/or structural conservation of TRV 16K. Confocal laser scanning microscopy of transiently expressed 16K fusions to red fluorescent protein (RFP) revealed a predominant cytoplasmic localization and, in addition, a nuclear localization. In contrast, fusions of RFP with the N-terminal region of 16K localized exclusively to the cytoplasm, whereas fusions between RFP and the C-terminal region of 16K displayed an exclusive nuclear localization. Further analysis of 16K-derived peptide fusions demonstrated that the 16K C-terminal region contained at least two functional bipartite nuclear localization signals which were independently capable of nuclear targeting.


Improvement of the gas cluster ion beam-(GCIB)-based molecular secondary ion mass spectroscopy (SIMS) depth profile with O2(+) cosputtering.
Over the last decade, cluster ion beams have displayed their capability to analyze organic materials and biological specimens. Compared with atomic ion beams, cluster ion beams non-linearly enhance the sputter yield, suppress damage accumulation and generate high mass fragments during sputtering. These properties allow successful Secondary Ion Mass Spectroscopy (SIMS) analysis of soft materials beyond the static limit. Because the intensity of high mass molecular ions is intrinsically low, enhancing the intensity of these secondary ions while preserving the sample in its original state is the key to highly sensitive molecular depth profiles. In this work, bulk poly(ethylene terephthalate) (PET) was used as a model material and analyzed using Time-of-Flight SIMS (ToF-SIMS) with a pulsed Bi3(2+) primary ion. The optimized hardware of a 10 kV Ar2500(+) Gas Cluster Ion Beam (GCIB) with a low kinetic energy (200-500 V) oxygen ion (O2(+)) as a cosputter beam was employed for generating depth profiles and for examining the effect of beam parameters. The results were then quantitatively analyzed using an established erosion model. It was found that the ion intensity of the PET monomer ([M + H](+)) and its large molecular fragment ([M - C2H4O + H](+)) steadily declined during single GCIB sputtering, with distortion of the distribution information. However, under an optimized GCIB-O2(+) cosputter, the secondary ion intensity quickly reached a steady state and retained >95% intensity with respect to the pristine surface, although the damage cross-section was larger than that of single GCIB sputtering. This improvement was due to the oxidation of molecules and the formation of -OH groups that serve as proton donors to particles emitted from the surface. As a result, the ionization yield was enhanced and damage to the chemical structure was masked. Although O2(+) is known to alter the chemical structure and cause damage accumulation, the concurrently used GCIB could sufficiently remove the surface layer and allow the damage to be masked by the enhanced ionization yield when the ion-solid interaction volume was kept shallow with a low O2(+) energy. This low O2(+) energy (200 V) cosputtering also produced a smoother surface than a single GCIB. Because the oxidized species were produced by O2(+) and removed by GCIB simultaneously, a sufficiently high O2(+) current density was required to produce adequate enhancements. Therefore, it was found that 10 kV with 2 × 10(-6) A per cm(2) Ar2500(+) and 200 V with 3.2 × 10(-4) A per cm(2) O2(+) produced the best profile.


[The pulmonary microangiopathy in patients with tuberculosis coexisting with diabetes mellitus].
There were done histologic, electron-microscopic investigations of the lungs preparations in 47 patients with destructive tuberculosis (DT) and diabetes mellitus, the operations were performed, the signs of the disseminated microangiopathy were revealed. This complication negatively influence the DT course before and after the operation, and the final result of operation also.


Feature-based registration of retinal images.
Registration of retinal images taken at different times frequently is required to measure changes caused by disease or to document retinal location of visual stimuli. Cross-correlation has been used previously for such registration, but it is computationally intensive. We have modified a faster algorithm, sequential similarity detection (SSD), to use only the portion of the template that contains retinal vessels. When compared to standard SSD and cross-correlation, this modification improves the reliability of detection for a variety of retinal imaging modalities. The improved reliability enables implementation of a two-stage registration strategy that further decreases the amount of computation and increases the speed of registration.


Cross-inhibitory activity of cereal protein inhibitors against alpha-amylases and xylanases.
The purification and characterisation of a xylanase inhibitor (XIP-I) from wheat was reported previously. In our current work, XIP-I is also demonstrated to have the capacity to inhibit the two barley alpha-amylase isozymes (AMY1 and AMY2). XIP-I completely inhibited the activity of AMY1 and AMY2 towards insoluble Blue Starch and a soluble hepta-oligosaccharide derivative. A ternary complex was formed between insoluble starch, a catalytically inactive mutant of AMY1 (D180A), and XIP-I, suggesting that the substrate-XIP-I interaction is necessary for inhibition of barley alpha-amylases. K(i) values for alpha-amylase inhibition, however, could not be calculated due to the nonlinear nature of the inhibition pattern. Furthermore, surface plasmon resonance and gel electrophoresis did not indicate interaction between XIP-I and the alpha-amylases. The inhibition was abolished by CaCl(2), indicating that the driving force for the interaction is different from that of complexation between the barley alpha-amylase/subtilisin inhibitor (BASI) and AMY2. This is the first report of a proteinaceous inhibitor of AMY1. BASI, in addition, was demonstrated to partially inhibit the endo-1,4-beta-D-xylanase from Aspergillus niger (XylA) of glycoside hydrolase family 11. Taken together, the data demonstrate for the first time the dual target enzyme specificity of BASI and XIP-I inhibitors for xylanase and alpha-amylase.


Endoscopic Endonasal Approach to the Optic Canal: Anatomic Considerations and Surgical Relevance.
Increasing use of endoscopic endonasal surgery for suprasellar lesions with extension into the optic canal (OC) has necessitated a better endonasal description of the OC. To identify the osseous OC transcranially and then investigate its anatomic relationship to the key endonasal intrasphenoidal landmarks. We also aimed to determine and describe the technical nuances for safely opening the falciform ligament and intracanalicular dura (surrounding the optic nerve [ON]) endonasally. Ten fresh human head silicon-injected specimens underwent an endoscopic transtuberculum/transplanum approach followed by 2-piece orbitozygomatic craniotomy to allow identification of 20 OCs. After completing up to 270° of endonasal bony decompression of the OC, a dural incision started at the sella and continued superiorly across the superior intercavernous sinus. Subsequently the dural opening was extended anterolaterally across the dura of the prechiasmatic sulcus, limbus sphenoidale, and planum. Endonasally, the length of the osseous OC was approximately 6 mm and equivalent to the length of the lateral opticocarotid recess, as measured anteroposteriorly. The ophthalmic artery arose from the supraclinoidal carotid artery at approximately 2.5 mm from the medial osseous OC entrance. Transcranial correlation of the endonasal dural incision confirmed medial detachment of the falciform ligament and exposure of the preforaminal ON. The lateral opticocarotid recess allows distinction of the preforaminal ON, roofed by the falciform ligament from the intracanalicular segment in the osseous OC. This facilitates the preoperative surgical strategy regarding the extent of OC decompression and dural opening. Extensive endonasal decompression of the OC and division of the falciform ligament is feasible.


The Rtf1 component of the Paf1 transcriptional elongation complex is required for ubiquitination of histone H2B.
In yeast cells, the Rtf1 and Paf1 components of the Paf1 transcriptional elongation complex are important for recruitment of Set1, the histone H3-lysine 4 (H3-Lys4) methylase, to a highly localized domain at the 5' portion of active mRNA coding regions. Here, we show that Rtf1 is essential for global methylation of H3-Lys4 and H3-Lys79, but not H3-Lys36. This role of Rtf1 resembles that of Rad6, which mediates ubiquitination of histone H2B at lysine 123. Indeed, Rtf1 is required for H2B ubiquitination, suggesting that its effects on H3-Lys4 and H3-Lys79 methylation are an indirect consequence of its effect on H2B ubiquitination. Rtf1 is important for telomeric silencing, with loss of H3-Lys4 and H3-Lys79 methylation synergistically reducing Sir2 association with telomeric DNA. Dot1, the H3-Lys79 methylase, associates with transcriptionally active genes, but unlike the association of Set1 and Set2 (the H3-Lys36 methylase), this association is largely independent of Rtf1. We suggest that Rtf1 affects genome-wide ubiquitination of H2B by a mechanism that is distinct from its function as a transcriptional elongation factor.


Abdominal Hysterectomy: Reduced Risk of Surgical Site Infection Associated with Robotic and Laparoscopic Technique.
Hysterectomy is one of the most common procedures performed in the United States. New techniques utilizing laparoscopic and robotic technology are becoming increasingly common. It is unknown if these minimally invasive surgical techniques alter the risk of surgical site infections (SSI). We performed a retrospective review of all patients undergoing abdominal hysterectomy at our institution between January 2011 and June 2013. International Classification of Diseases, Ninth edition (ICD-9) codes and chart review were used to identify patients undergoing hysterectomy by open, laparoscopic, or robotic approach and to identify patients who developed SSI subsequently. Chi-square and analysis of variance (ANOVA) tests were used to identify univariate risk factors and logistic regression was used to perform multivariable analysis. During this time period, 986 patients were identified who had undergone abdominal hysterectomy, with 433 receiving open technique (44%), 116 laparoscopic (12%), 407 robotic (41%), and 30 cases that were converted from minimally invasive to open (3%). Patients undergoing laparoscopic-assisted hysterectomy were significantly younger and had lower body mass index (BMI) and American Society of Anesthesiologists (ASA) scores than those undergoing open or robotic hysterectomy. There were no significant differences between patients undergoing open versus robotic hysterectomy. The post-operative hospital stay was significantly longer for open procedures compared with those using laparoscopic or robotic techniques (5.1, 1.7, and 1.6 d, respectively; p<0.0001). The overall rate of SSI after all hysterectomy procedures was 4.2%. More SSI occurred in open cases (6.5%) than laparoscopic (0%) or robotic (2.2%) (p<0.0001). Cases converted to open also had an increased rate of SSI (13.3%). In both univariate and multivariable analyses, open technique, wound class of III/IV, age greater than 75 y, and morbid obesity were all associated with increased risk of SSI. Laparoscopic and robotic hysterectomies were associated with a significantly lower risk of SSI and shorter hospital stays. Body mass index, advanced age, and wound class were also independent risk factors for SSI.


Response of soil dissolved organic matter to microplastic addition in Chinese loess soil.
Plastic debris is accumulating in agricultural land due to the increased use of plastic mulches, which is causing serious environmental problems, especially for biochemical and physical properties of the soil. Dissolved organic matter (DOM) plays a central role in driving soil biogeochemistry, but little information is available on the effects of plastic residues, especially microplastic, on soil DOM. We conducted a soil-incubation experiment in a climate-controlled chamber with three levels of microplastic added to loess soil collected from the Loess Plateau in China: 0% (control, CK), 7% (M1) and 28% (M2) (w/w). We analysed the soil contents of dissolved organic carbon (DOC), dissolved organic nitrogen (DON), NH4+, NO3-, dissolved organic phosphorus (DOP), and PO43- and the activities of fluorescein diacetate hydrolase (FDAse) and phenol oxidase. The higher level of microplastic addition significantly increased the nutrient contents of the DOM solution. The lower level of addition had no significant effect on the DOM solution during the first seven days, but the rate of DOM decomposition decreased in M1 between days 7 and 30, which increased the nutrient contents. The microplastic facilitated the accumulation of high-molecular-weight humic-like material between days 7 and 30. The DOM solutions were mainly comprised of high-molecular-weight humic-like material in CK and M1 and of high-molecular-weight humic-like material and tyrosine-like material in M2. The Microplastic stimulated the activities of both enzymes. Microplastic addition thus stimulated enzymatic activity, activated pools of organic C, N, and P, and was beneficial for the accumulation of dissolved organic C, N and P.


Improving efficiency in meat production.
Selective breeding and improved nutritional management over the past 20-30 years has resulted in dramatic improvements in growth efficiency for pigs and poultry, particularly lean tissue growth. However, this has been achieved using high-quality feed ingredients, such as wheat and soya that are also used for human consumption and more recently biofuels production. Ruminants on the other hand are less efficient, but are normally fed poorer quality ingredients that cannot be digested by human subjects, such as grass or silage. The challenges therefore are to: (i) maintain the current efficiency of growth of pigs and poultry, but using more ingredients not needed to feed the increasing human population or for the production of biofuels; (ii) improve the efficiency of growth in ruminants; (iii) at the same time produce animal products (meat, milk and eggs) of equal or improved quality. This review will describe the use of: (a) enzyme additives for animal feeds, to improve feed digestibility; (b) known growth promoting agents, such as growth hormone, β-agonists and anabolic steroids, currently banned in the European Union but used in other parts of the world; (c) recent transcriptomic studies into molecular mechanisms for improved growth efficiency via low residual feed intake. In doing so, the use of genetic manipulation in animals will also be discussed.


Estimation and interpretation of fermentation in the gut: coupling results from a 24 h batch in vitro system with fecal measurements from a human intervention feeding study using fructo-oligosaccharides, inulin, gum acacia, and pea fiber.
Gut bacteria ferment fiber at different rates to primarily short chain fatty acids (SCFA) and gas while proteins are metabolized to SCFA, branched chain fatty acids (BCFA), gas, and undesirable metabolites. Large volumes of gas produced in vivo may contribute to bloating and flatulence in an individual. The objectives of this trial were to (1) compare the in vitro fermentation profiles of fructo-oligosaccharides (FOS), inulin, gum acacia, and pea fiber alone or blended using a 24 h batch model and (2) relate these findings to a human study that fed enteral formula fortified with fiber blend (FB) or no fiber (FF). The in vitro fermentation of the fiber blend resulted in a delayed pH decrease and gas and SCFA production compared to the FOS and inulin. Human samples had higher SCFA on FB compared to FF (p = 0.029). BCFA were not different between formulas. By using a blend of fibers, we observed a slower fermentation in vitro but still increased fecal SCFA when fed to human subjects.


A Method of Locating the Dehiscence during Middle Fossa Approach for Superior Semicircular Canal Dehiscence Surgery.
Objectives Superior semicircular canal dehiscence (SSCD) results from a defect in the middle cranial fossa floor. One challenge during SSCD repair is the lack of a consistent landmark. This study proposes a reference point above the external auditory canal at the level of the zygoma as the inferior craniectomy edge during surgery. Design This is a retrospective review of patients with SSCD. Setting/Participants A total of 72 cases of SSCD in 60 patients were repaired via a middle fossa approach at a single institution. Main Outcome Measures The distance from the proposed reference point to the dehiscence was statistically analyzed using Shapiro-Wilk's goodness-of-fit test and Student's t -test. Results Average distance for all patients was 28.84 ± 2.22 mm (range: 22.96-33.43). Average distance for females was 29.08 mm (range: 24.56-33.43) versus 28.26 mm (range: 22.96-32.36) for males. There was no difference in distance by sex ( p = 0.174). The distance measurements followed a normal distribution with 95% of the patients between 24.49 and 33.10 mm. Conclusion This study analyzed a potential reference point during a middle fossa approach for SSCD surgery. The distance from this reference point to the SSCD was found to be consistent and may serve as a readily identifiable landmark in localizing the dehiscence.


RIblast: an ultrafast RNA-RNA interaction prediction system based on a seed-and-extension approach.
LncRNAs play important roles in various biological processes. Although more than 58 000 human lncRNA genes have been discovered, most known lncRNAs are still poorly characterized. One approach to understanding the functions of lncRNAs is the detection of the interacting RNA target of each lncRNA. Because experimental detections of comprehensive lncRNA-RNA interactions are difficult, computational prediction of lncRNA-RNA interactions is an indispensable technique. However, the high computational costs of existing RNA-RNA interaction prediction tools prevent their application to large-scale lncRNA datasets. Here, we present 'RIblast', an ultrafast RNA-RNA interaction prediction method based on the seed-and-extension approach. RIblast discovers seed regions using suffix arrays and subsequently extends seed regions based on an RNA secondary structure energy model. Computational experiments indicate that RIblast achieves a level of prediction accuracy similar to those of existing programs, but at speeds over 64 times faster than existing programs. The source code of RIblast is freely available at https://github.com/fukunagatsu/RIblast . firstname.lastname@example.com or email@example.com. Supplementary data are available at Bioinformatics online.


[An experimental study of the morphogenetic properties of several phenols inducing phenocopies of vitiligo].
In an experiment with black guinea pigs a phenocopy of vitiligo was obtained by means of the exposure of the skin to the action of p-tert-butyl-phenol (PTBP) or to catechol (C). Two other compounds: 2,2-dihydroxy-diphenylpropane (DDP) and 2, 4, 6-tri-tert-butyl-phenol (TTBP) exerted a hypopigmentary effect. PTBP and C depigmented the skin but caused no preceding inflammation. Spots of depigmented (white) skin and hairs were surrounded by a zone of hyperpigmentation. Leukoderma proved to be stable, in some cases irreversible, and exhibited a tendency to progressing and spontaneous dissemination. In the experiments with Drosophila melanogaster both PTBP and C were observed to possess a morphogenetic capacity: they induced a change in the puparium colour (C), as well as of the colour of body and wings of adult flies, inducing also disturbances of wing development practically in 100% of individuals to which these substances were administrated with food at the larval stage. None of the four preparations tested exerted any mutagenic effect in cells of imaginal buds of Drosophila melanogaster.


Multipotential human adipose-derived stromal stem cells exhibit a perivascular phenotype in vitro and in vivo.
Mesenchymal stem-like cells identified in different tissues reside in a perivascular niche. In the present study, we investigated the putative niche of adipose-derived stromal/stem cells (ASCs) using markers, associated with mesenchymal and perivascular cells, including STRO-1, CD146, and 3G5. Immunofluorescence staining of human adipose tissue sections, revealed that STRO-1 and 3G5 co-localized with CD146 to the perivascular regions of blood vessels. FACS was used to determine the capacity of the CD146, 3G5, and STRO-1 specific monoclonal antibodies to isolate clonogenic ASCs from disassociated human adipose tissue. Clonogenic fibroblastic colonies (CFU-F) were found to be enriched in those cell fractions selected with either STRO-1, CD146, or 3G5. Flow cytometric analysis revealed that cultured ASCs exhibited similar phenotypic profiles in relation to their expression of cell surface markers associated with stromal cells (CD44, CD90, CD105, CD106, CD146, CD166, STRO-1, alkaline phosphatase), endothelial cells (CD31, CD105, CD106, CD146, CD166), haematopoietic cells (CD14, CD31, CD45), and perivascular cells (3G5, STRO-1, CD146). The immunoselected ASCs populations maintained their characteristic multipotential properties as shown by their capacity to form Alizarin Red positive mineralized deposits, Oil Red O positive lipid droplets, and Alcian Blue positive proteoglycan-rich matrix in vitro. Furthermore, ASCs cultures established from either STRO-1, 3G5, or CD146 selected cell populations, were all capable of forming ectopic bone when transplanted subcutaneously into NOD/SCID mice. The findings presented here, describe a multipotential stem cell population within adult human adipose tissue, which appear to be intimately associated with perivascular cells surrounding the blood vessels.


Quality-adjusted cost of care: a meaningful way to measure growth in innovation cost versus the value of health gains.
Technology drives both health care spending and health improvement. Yet policy makers rarely see measures of cost growth that account for both effects. To fill this gap, we present the quality-adjusted cost of care, which illustrates cost growth net of growth in the value of health improvements, measured as survival gains multiplied by the value of survival. We applied the quality-adjusted cost of care to two cases. For colorectal cancer, drug cost per patient increased by $34,493 between 1998 and 2005 as a result of new drug launches, but value from offsetting health improvements netted a modest $1,377 increase in quality-adjusted cost of care. For multiple myeloma, new therapies increased treatment cost by $72,937 between 2004 and 2009, but offsetting health benefits lowered overall quality-adjusted cost of care by $67,863. However, patients with multiple myeloma on established first-line therapies saw costs rise without corresponding benefits. All three examples document rapid cost growth, but they provide starkly different answers to the question of whether society got what it paid for.


Structural and Barrier Properties of Compatibilized PE/PA6 Multinanolayer Films.
The barrier performance and structural lightening of organic materials are increasingly desired and constitute a major challenge for manufacturers, particularly for transport and packaging. A promising technique which tends to emerge in recent years is that of multinanolayer coextrusion. The advantage is that it can produce multilayers made of thousands of very thin layers, leading to new properties due to crystalline morphology changes induced by confinement. This paper is focusing on the study of multinanolayered films with alternated polyethylene (PE), compatibilizer (PEgMA) and polyamide 6 (PA6) layers and made by a forced assembly coextrusion process equipped with layer multiplying elements (LME). PE/PA6 multilayer films consisting of 5 to 2049 layers (respectively 0 to 9 LME) were successfully obtained with well-organized multilayered structure. The evolution of the morphology and the microstructure of these two semi-crystalline polymers, when the thickness of each polymer layer decreases from micro-scale to nano-scale, was correlated to the water and gas transport properties of the PE/PA multilayers. The expected improvement of barrier properties was limited due to the on-edge orientation of crystals in very thin PE and PA6 layers. Despite this change of crystalline morphology, a slight improvement of the gas barrier properties was shown by comparing experimental results with permeabilities predicted on the basis of a serial model developed by considering a PE/PA6 interphase. This interphase observed by TEM images and the on-edge crystal orientation in multilayers were evidenced from mechanical properties showing an increase of the stiffness and the strength.


Failure of highly purified lysyl hydroxylase to hydroxylate lysyl residues in the non-helical regions of collagen.
The activity of highly purified lysyl hydroxylase towards lysyl residues within both the helical and the N-terminal non-helical telopeptide regions of chick type I collagen has been examined. The peptides alpha 1(I)-CB1 and alpha 2(I)-CB1, isolated from protocollagen following CNBr digestion and containing the N-terminal telopeptidyl lysyl residues, failed themselves to act as substrates. With protocollagen as substrate, analysis of products obtained following bacterial collagenase digestion of the reaction mixture showed that overall 37% hydroxylation of lysyl residues within the helical region of collagen had been obtained, which may be maximal. No hydroxylation, however, of the single lysyl residue in either alpha 1(I)-CB1 or alpha 2(I)-CB1, isolated following CNBr digestion of the reaction mixture, was observed, despite the known susceptibility of these residues to hydroxylation. These findings provide strong circumstantial evidence for the suggestion that a lysyl hydroxylase specific for the telopeptidyl residues and distinct from that active towards lysyl residues in the helical portion of the molecule may exist [Barnes, Constable, Morton & Royce (1974) Biochem. J. 139, 461-468].


Cognitive process scores associated with self-reported behavioral dysfunction on the Frontal Systems Behavior Scale (FrSBe) in chronic traumatic brain injury.
Introduction: Individuals with traumatic brain injury (TBI) often experience long-term cognitive impairment that affects daily functioning, but neuropsychological tests are often not associated with functional outcomes in chronic TBI. To identify better neuropsychological test predictors of functional outcomes, we identified process-oriented neuropsychological test scores associated with behavioral dysfunction in chronic TBI.Method: Adults (N = 65, age = 49.9 years, education = 14.6 years, 80% male, 97% white) with a > 6-month (median = 55 months) history of TBI (31% complicated mild, 20% moderate, 49% severe) completed the self-reported Frontal Systems Behavior Scale (FrSBe) and a battery of neuropsychological tests. Cognitive scores significantly associated with FrSBe domain scores (Apathy, Disinhibition, Executive Dysfunction) were included in hierarchical linear regression analyses.Results: Pearson correlations were significant (|r| = .25 - .41, p < .05) between FrSBe Disinhibition and Executive Dysfunction domains and numerous cognitive scores, particularly cognitive process scores, including California Verbal Learning Test II (CVLT-II) and Letter (FAS) Fluency Intrusions and Repetitions. In the regression analyses, gender, injury severity, premorbid intelligence, time since injury, self-awareness, and depression status accounted for less than 1% (Disinhibition) and 6.1% (Executive Dysfunction) of the total adjusted variance in respective FrSBe domains. Cognitive process scores were the strongest factors associated with behavioral dysfunction, explaining an additional 22.9% of the total adjusted variance in Disinhibition [F(3, 50) = 6.28, p = .001]. For Executive Dysfunction, traditional scores explained 2.6% added variance on top of the base model, with cognitive process scores explaining an additional 17.2% [F(4, 48) = 6.79, p = .003] of the total adjusted variance.Conclusions: Less frequently utilized cognitive process scores from well-known neuropsychological tests may be better predictors of behavioral dysfunction in chronic TBI, as measured by the FrSBe. Future prospective study is required to validate these findings and test the sensitivity and specificity of these cognitive process scores for identifying risk for behavioral dysfunction.


Low density lipoprotein receptor and mRNA expression in human colorectal cancer.
We have previously show that, 63.3% of colorectal cancers did not express the Low Density Lipoprotein Receptor (LDLR). We now report the findings of a study on the expression of LDLR and its mRNA in neoplastic tissue specimens of human colorectal cancer (CRC), carried out to verify whether the absence of the LDLR in these tumours is reflected by the absence of its transcript. 32 patients (10 females and 22 males) operated for CRC were included in the study. The LDLR levels were evaluated by immunoenzymatic assay. The LDLR-mRNA, reverse-transcribed and then amplified by polymerase chain reaction, was detected by chromatography. The LDLR protein was present in 12 out of the 32 patients. LDLR-mRNA expression was detected in 17 out of the 32 patients. The absence of the LDLR protein was reflected by the absence of its transcript in 13 out of the 20 tumours; The LDLR mRNA levels were significantly higher in the tumours that did not expres LDLR. The variable expression of the LDLR protein and LDLR mRNA in CRC detected in this study suggests the possibility that different therapeutic strategies may be indicated for these tumours.


Simultaneous screening and target analytical approach by gas chromatography-quadrupole-mass spectrometry for pesticide residues in fruits and vegetables.
A full-scan GC/quadrupole/MS method has been developed to perform large-scale screenings of pesticides and simultaneous quantification of 95 target compounds in a single run of 21 min. The screening method was performed by using a deconvolution of the spectrum of the full-scan data files acquired under a retention time locked method. The identification performance of the screening method was evaluated in eight different food matrixes at three different concentrations. The system was equipped with a programmable temperature vaporizing inlet, allowing 10 microL injections. The LOQ in the full-scan mode and linearity were studied for four different matrixes. Correlation coefficients > 0.99 were achieved in all cases, and the LOD was < 20 microg/kg for 80% of the studied pesticides. Maintenance of the system was reduced by the use of a QuickSwap device that provided backflush capabilities by reversing column flow immediately after elution of the last compound of interest. The combined screening and target method was used in the analysis of more than 100 food samples, including a carrot sample from the European Proficiency Test FV 10, with good results.


Beyond inhibition: a dual-process perspective to renew the exploration of binge drinking.
Binge drinking is a widespread alcohol-consumption pattern in youth and is linked to cognitive consequences, mostly for executive functions. However, other crucial factors remain less explored in binge drinking and notably the emotional-automatic processes. Dual-process model postulates that addictive disorders are not only due to impaired reflective system (involved in deliberate behaviors), but rather to an imbalance between under-activated reflective system and over-activated affective-automatic one (involved in impulsive behaviors). This proposal has been confirmed in alcohol-dependence, but has not been tested in binge drinking. The observation of comparable impairments in binge drinking and alcohol-dependence led to the "continuum hypothesis," suggesting similar deficits across different alcohol-related disorders. In this perspective, applying the dual-process model to binge drinking might renew the understanding of this continuum hypothesis. A three-axes research agenda will be proposed, exploring: (1) the affective-automatic system in binge drinking; (2) the systems' interactions and imbalance in binge drinking; (3) the evolution of this imbalance in the transition between binge drinking and alcohol-dependence.


The Arp2/3 Complex Is Essential for Distinct Stages of Spine Synapse Maturation, Including Synapse Unsilencing.
Dendritic filopodia are actin-rich structures that are thought to contribute to early spine synapse formation; however, the actin regulatory proteins important for early synaptogenesis are poorly defined. Using organotypic hippocampal slice cultures and primary neuron hippocampal cultures from Arp2/3 conditional knock-out mice, we analyze the roles of the Arp2/3 complex, an actin regulator that creates branched actin networks, and demonstrate it is essential for distinct stages of both structural and functional maturation of excitatory spine synapses. Our data show that initially the Arp2/3 complex inhibits the formation of dendritic filopodia but that later during development, the Arp2/3 complex drives the morphological maturation from filopodia to typical spine morphology. Furthermore, we demonstrate that although the Arp2/3 complex is not required for key spine maturation steps, such as presynaptic contact and recruitment of MAGUK (membrane-associated guanylate kinase) scaffolding proteins or NMDA receptors, it is necessary for the recruitment of AMPA receptors. This latter process, also known as synapse unsilencing, is a final and essential step in the neurodevelopment of excitatory postsynaptic synaptogenesis, setting the stage for neuronal interconnectivity. These findings provide the first evidence that the Arp2/3 complex is directly involved in functional maturation of dendritic spines during the developmental period of spinogenesis. Excitatory spine synapse formation (spinogenesis) is a poorly understood yet pivotal period of neurodevelopment that occurs within 2-3 weeks after birth. Neurodevelopmental disorders such as intellectual disability and autism are characterized by abnormal spine structure, which may arise from abnormal excitatory synaptogenesis. The initial stage of spinogenesis is thought to begin with the emergence of actin-rich dendritic filopodia that initiate contact with presynaptic axonal boutons. However, it remains enigmatic how actin cytoskeletal regulation directs dendritic filopodial emergence or their subsequent maturation into dendritic spines during development and on into adulthood. In this study, we provide the first evidence that the Arp2/3 complex, a key actin nucleator, is involved in distinct stages of spine formation and is required for synapse unsilencing.