Measuring anger in patients experiencing chronic pain - A systematic review.
Anger is prevalent in chronic pain and has been associated with pain perception, disability, behavior and treatment outcome. Objectives were (1) to survey in the context of chronic pain the application (and omission) of validated anger self-report instruments, (2) to discuss the instruments found in the context of emotion theories and (3) to identify a possible instrument preference. A systematic search of textbooks and review articles was first performed on validated instruments designed to measure the cognitive, the motivational and the subjective feeling component of anger. Thereafter, a systematic review aimed at finding chronic pain studies from 2005 to 2019 reporting on these instruments. Textbooks and reviews listed 16 validated self-report anger measurement instruments. 28 papers applying four of these were identified and two new instruments were additionally detected. The State-Trait Anger Expression (STAXI) and its precursors were most commonly used. Studies on chronic low back pain patients prevailed. In conclusion, anger in chronic pain patients is reliably measurable at low cost with self-report tools. The STAXI-II qualifies best for this purpose based on its extensive validation history. The majority of instruments lack sufficient theoretical and psychometric adequacy. A more detailed exploration of the cognitive anger component in chronic pain patients in future research is recommended.


TGF-β activation and function in immunity.
The cytokine TGF-β plays an integral role in regulating immune responses. TGF-β has pleiotropic effects on adaptive immunity, especially in the regulation of effector and regulatory CD4(+) T cell responses. Many immune and nonimmune cells can produce TGF-β, but it is always produced as an inactive complex that must be activated to exert functional effects. Thus, activation of latent TGF-β provides a crucial layer of regulation that controls TGF-β function. In this review, we highlight some of the important functional roles for TGF-β in immunity, focusing on its context-specific roles in either dampening or promoting T cell responses. We also describe how activation of TGF-β controls its function in the immune system, with a focus on the key roles for members of the integrin family in this process.


One-pot stapling of interchain disulfides of antibodies using an isobutylene motif.
Monoclonal antibodies have emerged as an important class of therapeutics in oncological and autoimmune diseases due to their several attractive properties, such as high binding affinity and specificity. However, it has recently become clear that antibodies recovered from serum show a significantly decreased potency owing to various reasons, including deamidation, oxidation, fragment antigen binding (Fab) exchange, and disulfide shuffling. Fab exchange and disulfide shuffling result because of the instability of disulfides in serum. Herein, we reported a 'one-pot' stapling strategy using isobutylene motifs to stabilise the interchain disulfides of antibodies. This general method was applied to a Fab fragment of the anti-HER2 antibody. The stapled Fab was completely stable in the presence of biological thiols. The approach was further applied to two different full-length IgGs, trastuzumab and rituximab, under mild and biocompatible conditions. The binding affinity of the antibody was enhanced, relative to its native form, after being stapled. The stapled structure maintained its effector functions and behaved similarly to its native form in vivo. This work provides a straightforward and scalable method for the stabilisation of antibodies in various formats.


Localization of a circadian pacemaker in the eye of a mollusc, bulla.
The eye of the marine mollusc Bulla contains a circadian pacemaker which, along with critical entrainment pathways, is located among a small group of neurons at the base of the retina. Long-term intracellular recording from cells of the organized photoreceptor layer, which constitutes most of the retinal volume, indicates that these cells are not involved in generating the rhythm since rhythmic changes in membrane potential were not observed. In addition, surgical removal of the entire photoreceptor layer does not alter the period of the circadian rhythm and does not prevent phase shifts by light pulses.


Parental encouragement of dieting promotes daughters' early dieting.
Dieting to lose weight is common among female adolescents. This research investigated the association between maternal and paternal encouragement to diet and their daughters' self-reported "early dieting" (prior to age 11 y) and adolescent dieting (between 11 y and 15 y), and how parental encouragement to diet is related to changes in daughters' BMI percentiles. Participants in this study were 174 non-Hispanic white girls and their parents, assessed when daughters were 9-, 11-, 13-, and 15 y. The Parent Encouragement of Child Weight Loss Scale was used to measure encouragement to diet. Logistic regression was used to examine the relationship between parental encouragement to diet and daughters' reports of dieting by 11 y and by 15 y, adjusting for daughters' weight status at baseline. Compared with girls whose mothers didn't encourage dieting, girls who were encouraged to diet were twice as likely to diet by 11 y; girls who were encouraged by their fathers were also twice as likely to diet by 11 y. Girls who were encouraged to diet by both parents were 8 times more likely to report early dieting than girls who were not. Neither maternal nor paternal encouragement predicted the emergence of dieting during adolescence. Girls who dieted and had parental encouragement to do so had increases in BMI percentile from 9 y to 15 y. Findings reveal that parental encouragement to diet may be counterproductive and that parents need alternative approaches to promote healthy patterns of intake and growth among young girls.


Increase in PKCα Activity during Heart Failure Despite the Stimulation of PKCα Braking Mechanism.
Rationale: Heart failure (HF) is marked by dampened cardiac contractility. A mild therapeutic target that improves contractile function without desensitizing the β-adrenergic system during HF may improve cardiac contractility and potentially survival. Inhibiting protein kinase C α (PKCα) activity may fit the criteria of a therapeutic target with milder systemic effects that still boosts contractility in HF patients. PKCα activity has been observed to increase during HF. This increase in PKCα activity is perplexing because it is also accompanied by up-regulation of a molecular braking mechanism. Objective: I aim to explore how PKCα activity can be increased and maintained during HF despite the presence of a molecular braking mechanism. Methods and Results: Using a computational approach, I show that the local diacylglycerol (DAG) signaling is regulated through a two-compartment signaling system in cardiomyocytes. These results imply that after massive myocardial infarction (MI), local homeostasis of DAG signaling is disrupted. The loss of this balance leads to prolonged activation of PKCα, a key molecular target linked to LV remodeling and dysfunctional filling and ejection in the mammalian heart. This study also proposes an explanation for how DAG homeostasis is regulated during normal systolic and diastolic cardiac function. Conclusions: I developed a novel two-compartment computational model for regulating DAG homeostasis during Ang II-induced heart failure. This model provides a promising tool with which to study mechanisms of DAG signaling regulation during heart failure. The model can also aid in identification of novel therapeutic targets with the aim of improving the quality of life for heart failure patients.


A conservative single visit reattachment of fractured crown fragment.
Injury of anterior teeth is a relatively common event that mainly affects children and adolescents. Dentists are confronted with managing dental trauma and restoring fractured teeth on a regular basis. Hence the techniques that speed and simplify the treatment, restore esthetics and improve long term success rate are considered of potential value. If an intact tooth fragment is present after trauma, immediate attachment of the incisal edge is a conservative yet, simple and aesthetic alternative. Fracture reattachment possesses challenging conservative and economically viable procedure within a single visit. The authors wuld report a case of fractured maxillary central incisor using fragment reattachment.


Predicting alcohol use disorder remission: a longitudinal multimodal multi-featured machine learning approach.
Predictive models for recovering from alcohol use disorder (AUD) and identifying related predisposition biomarkers can have a tremendous impact on addiction treatment outcomes and cost reduction. Our sample (N = 1376) included individuals of European (EA) and African (AA) ancestry from the Collaborative Study on the Genetics of Alcoholism (COGA) who were initially assessed as having AUD (DSM-5) and reassessed years later as either having AUD or in remission. To predict this difference in AUD recovery status, we analyzed the initial data using multimodal, multi-features machine learning applications including EEG source-level functional brain connectivity, Polygenic Risk Scores (PRS), medications, and demographic information. Sex and ancestry age-matched stratified analyses were performed with supervised linear Support Vector Machine application and were calculated twice, once when the ancestry was defined by self-report and once defined by genetic data. Multifeatured prediction models achieved higher accuracy scores than models based on a single domain and higher scores in male models when the ancestry was based on genetic data. The AA male group model with PRS, EEG functional connectivity, marital and employment status features achieved the highest accuracy of 86.04%. Several discriminative features were identified, including collections of PRS related to neuroticism, depression, aggression, years of education, and alcohol consumption phenotypes. Other discriminated features included being married, employed, medication, lower default mode network and fusiform connectivity, and higher insula connectivity. Results highlight the importance of increasing genetic homogeneity of analyzed groups, identifying sex, and ancestry-specific features to increase prediction scores revealing biomarkers related to AUD remission.


Validation of an ultra-fast UPLC-UV method for the separation of antituberculosis tablets.
A simple method using ultra performance LC (UPLC) coupled with UV detection was developed and validated for the determination of antituberculosis drugs in combined dosage form, i. e. isoniazid (ISN), pyrazinamide (PYR) and rifampicin (RIF). Drugs were separated on a short column (2.1 mm x 50 mm) packed with 1.7 mum particles, using an elution gradient procedure. At 30 degrees C, less than 2 min was necessary for the complete separation of the three antituberculosis drugs, while the original USP method was performed in 15 min. Further improvements were obtained with the combination of UPLC and high temperature (up to 90 degrees C), namely HT-UPLC, which allows the application of higher mobile phase flow rates. Therefore, the separation of ISN, PYR and RIF was performed in less than 1 min. After validation (selectivity, trueness, precision and accuracy), both methods (UPLC and HT-UPLC) have proven suitable for the routine quality control analysis of antituberculosis drugs in combined dosage form. Additionally, a large number of samples per day can be analysed due to the short analysis times.


Bullying and victimization in early adolescence: relations to social information processing patterns.
There is a gap in the literature on the social information processing (SIP) patterns of adolescents exposed to victimization in school. Therefore, we examine the SIP patterns of young adolescents characterized by their teachers and by their own reports as victims, bullies, bullies/victims, and neither bullies nor victims. The 105 adolescents participating in this study were asked to respond to hypothetical social scenarios in which a protagonist is either rebuffed or provoked by peers. The scenarios were ambiguous in nature and thus could have been processed in different ways. Indeed, distinctive processing patterns were found for each of these groups: victims tended to avoid challenging social situations while expecting others to be purposefully hostile or ignoring; bullies tended to interpret others as purposefully hostile and stated their desire to retaliate; bullies/victims showed patterns more similar to those of the bullies than the victims; and those who were neither victims nor bullies tended to view the same challenging social situations as non hostile and more likely to end well for them. We conclude by discussing the theoretical and practical implications of these findings.


Preliminary experience with high-dose cisplatin, reduced glutathione and natural interferon-alpha in dacarbazine-resistant malignant melanoma.
The incidence of malignant melanoma is rapidly increasing in many countries, and when this disease has reached advanced stages, standard therapies have little impact. Dacarbazine (DTIC) is the most effective chemotherapeutic agent with an overall response rate of 20-25%, but durable responses are uncommon. Interesting results with the use of cisplatin (CDDP) have been reported in DTIC-resistant melanoma. Moreover, malignant melanoma is an immunogenic tumor and a potential target for biological response modifier (BRM) therapies. The aim of the present study was to evaluate the efficacy and tolerability of a chemo-immunotherapeutic regimen including high-dose CDDP combined with glutathione (GSH) to limit platinum-related toxicity, and natura interferon-alpha (IFN-alpha) in patients with DTIC-resistant metastatic melanoma. The treatment schedule included GSH 1,500 mg/m2 i.v. and CDDP 40 mg/m2 i.v. for 4 consecutive days every 3 weeks, with a maximum of 6 courses, and IFN-alpha 3 MIU i.m. 3 times a week, continuative for a maximum of 12 months. Twelve patients were enrolled in this phase II trial. Accrual was stopped due to treatment-related toxicity. Ten patients were evaluable for response; there were 2 partial responses, lasting 5+ and 9+ months, respectively, and 2 cases of stable disease, lasting 3+ and 8+ months. None of these patients completed the therapeutic program due to treatment-related side effects. This regimen seems to be only partially active in DTIC-resistant metastatic melanoma. Hematologic and non-hematologic (nausea and vomiting, peripheral neurotoxicity, and asthenia) side effects are significant and GSH is not effective in limiting CDDP-related neurotoxicity in pretreated patients. Therefore, there is no indication to employ this regimen as second-line treatment in metastatic melanoma and these disappointing results highlight the urgent need for new therapeutic approaches.


nextPYP: a comprehensive and scalable platform for characterizing protein variability in situ using single-particle cryo-electron tomography.
Single-particle cryo-electron tomography is an emerging technique capable of determining the structure of proteins imaged within the native context of cells at molecular resolution. While high-throughput techniques for sample preparation and tilt-series acquisition are beginning to provide sufficient data to allow structural studies of proteins at physiological concentrations, the complex data analysis pipeline and the demanding storage and computational requirements pose major barriers for the development and broader adoption of this technology. Here, we present a scalable, end-to-end framework for single-particle cryo-electron tomography data analysis from on-the-fly pre-processing of tilt series to high-resolution refinement and classification, which allows efficient analysis and visualization of datasets with hundreds of tilt series and hundreds of thousands of particles. We validate our approach using in vitro and cellular datasets, demonstrating its effectiveness at achieving high-resolution and revealing conformational heterogeneity in situ. The framework is made available through an intuitive and easy-to-use computer application, nextPYP ( http://nextpyp.app ).


Voluntary breathing influences corticospinal excitability of nonrespiratory finger muscles.
The present study aimed to investigate neurophysiologic mechanisms mediating the newly discovered phenomenon of respiratory-motor interactions and to explore its potential clinical application for motor recovery. First, young and healthy subjects were instructed to breathe normally (NORM); to exhale (OUT) or inhale (IN) as fast as possible in a self-paced manner; or to voluntarily hold breath (HOLD). In experiment 1 (n = 14), transcranial magnetic stimulation (TMS) was applied during 10% maximal voluntary contraction (MVC) finger flexion force production or at rest. The motor-evoked potentials (MEPs) were recorded from flexor digitorum superficialis (FDS), extensor digitorum communis (EDC), and abductor digiti minimi (ADM) muscles. Similarly, in experiment 2 (n = 11), electrical stimulation (ES) was applied to FDS or EDC during the described four breathing conditions while subjects maintained 10%MVC of finger flexion or extension and at rest. In the exploratory clinical experiments (experiment 3), four patients with chronic neurological disorders (three strokes, one traumatic brain injury) received a 30-min session of breathing-controlled ES to the impaired EDC. In experiment 1, the EDC MEP magnitudes increased significantly during IN and OUT at both 10%MVC and rest; the FDS MEPs were enhanced only at 10%MVC, whereas the ADM MEP increased only during OUT, compared with NORM for both at rest and 10%MVC. No difference was found between NORM and HOLD for all three muscles. In experiment 2, when FDS was stimulated, force response was enhanced during both IN and OUT, but only at 10%MVC. When EDC was stimulated, force response increased at both 10%MVC and rest, only during IN, but not OUT. The averaged response latency was 83 ms for the finger extensors and 79 ms for the finger flexors. After a 30-min intervention of ES to EDC triggered by forced inspiration in experiment 3, we observed a significant reduction in finger flexor spasticity. The spasticity reduction lasted for ≥ 4 wk in all four patients. TMS and ES data, collectively, support the phenomenon that there is an overall respiration-related enhancement on the motor system, with a strong inspiration-finger extension coupling during voluntary breathing. As such, breathing-controlled electrical stimulation (i.e., stimulation to finger extensors delivered during the voluntary inspiratory phase) could be applied for enhancing finger extension strength and finger flexor spasticity reduction in poststroke patients.


Möbius syndrome in a neonate after mifepristone and misoprostol elective abortion failure.
To report a case of a child born with Möbius syndrome following exposure in utero to mifepristone and misoprostol for elective abortion. In the seventh week of pregnancy, a woman was administered mifepristone 600 mg and, 2 days later, misoprostol 400 mug for abortion. One month later, despite significant metrorrhagia, an ultrasound examination showed ongoing gestation. At 33 weeks and 3 days of gestation, the woman gave birth to a male with left facial palsy, microretrognathia, and axial hypotonia related to Möbius syndrome. Möbius syndrome is characterized by unilateral or bilateral palsy of the abducens (VI) and facial (VII) cranial nerves. Other cranial nerves (eg, the hypoglossal [XII]), craniofacial or orofacial anomalies, and limb malformations are often associated. The etiology of the Möbius syndrome remains largely unknown and probably involves multiple factors. The most likely etiological hypothesis is disruption of the developing vascular system, with transient ischemia (particularly in the vertebral arteries) and fetal hypoxia. A teratogenic cause of Möbius syndrome has been suggested. The critical period for the development of Möbius syndrome following teratogen exposure appears to be 5-8 weeks of gestation. To date, mifepristone alone does not appear to have induced Möbius syndrome. In contrast, oral or vaginal misoprostol administration can lead to a significant increase in Doppler-measured uterine artery resistance and may induce uterine contractions. If these occur during the critical embryonic period, they may cause flexion in the areas of the sixth and seventh cranial nerves and decreased blood flow. Ineffective use of mifepristone and misoprostol in the first trimester of pregnancy may be associated with a risk of Möbius syndrome, primarily due to misoprostol activity. Women with ongoing pregnancy after failed abortion with misoprostol administration should be informed of this risk.


Sexual Dimorphism in Innate Immunity.
Sexual dimorphisms account for differences in clinical manifestations or incidence of infectious or autoimmune diseases and malignancy between females and males. Females develop enhanced innate and adaptive immune responses than males and are less susceptible to many infections of bacterial, viral, parasitic, and fungal origin and malignancies but in contrast, they are more prone to develop autoimmune diseases. The higher susceptibility to infections in males is observed from birth to adulthood, suggesting that sex chromosomes and not sex hormones have a major role in sexual dimorphism in innate immunity. Sex-based regulation of immune responses ultimately contributes to age-related disease development and life expectancy. Differences between males and females have been described in the expression of pattern recognition receptors of the innate immune response and in the functional responses of phagocytes and antigen presenting cells. Different factors have been shown to account for the sex-based disparity in immune responses, including genetic factors and hormonal mediators, which contribute independently to dimorphism in the innate immune response. For instance, several genes encoding for innate immune molecules are located on the X chromosome. In addition, estrogen and/or testosterone have been reported to modulate the differentiation, maturation, lifespan, and effector functions of innate immune cells, including neutrophils, macrophages, natural killer cells, and dendritic cells. In this review, we will focus on differences between males and females in innate immunity, which represents the first line of defense against pathogens and plays a fundamental role in the activation, regulation, and orientation of the adaptive immune response.


Dual inhibition of CDK4/Rb and PI3K/AKT/mTOR pathways by ON123300 induces synthetic lethality in mantle cell lymphomas.
This study describes the characterization of a novel kinase inhibitor, ON123300, which inhibits CDK4/6 (cyclin-dependent kinases 4 and 6) and phosphatidylinositol 3 kinase-δ (PI3K-δ) and exhibits potent activity against mantle cell lymphomas (MCLs) both in vitro and in vivo. We examined the effects of PD0332991 and ON123300 on cell cycle progression, modulation of the retinoblastoma (Rb) and PI3K/AKT pathways, and the induction of apoptosis in MCL cell lines and patient-derived samples. When Granta 519 and Z138C cells were incubated with PD0332991 and ON123300, both compounds were equally efficient in their ability to inhibit the phosphorylation of Rb family proteins. However, only ON123300 inhibited the phosphorylation of proteins associated with the PI3K/AKT pathway. Cells treated with PD0332991 rapidly accumulated in the G0/G1 phase of cell cycle as a function of increasing concentration. Although ON123300-treated cells arrested similarly at lower concentrations, higher concentrations resulted in the induction of apoptosis, which was not observed in PD0332991-treated samples. Mouse xenograft assays also showed a strong inhibition of MCL tumor growth in ON123300-treated animals. Finally, treatment of ibrutinib-sensitive and -resistant patient-derived MCLs with ON123300 also triggered apoptosis and inhibition of the Rb and PI3K/AKT pathways, suggesting that this compound might be an effective agent in MCL, including ibrutinib-resistant forms of the disease.


Analysis of >100,000 Cardiovascular Surgeries Performed at the Heart Institute and a New Era of Outcomes.
The current challenge of cardiovascular surgery (CVS) is to improve the outcomes in increasingly severe patients. In this respect, continuous quality improvement (CQI) programs have had an impact on outcomes. To assess the evolution of the incidence and mortality due to CVS, as well as the current outcomes of the Hospital das Clínicas Heart Institute of the University of São Paulo Medical School (InCor-HCFMUSP). An outcome analysis of CVSs performed at the InCor, between January 1984 and June 2019. We observed the surgical volume and mortality rates in 5 time periods: 1st (1984-1989), 2nd (1990-1999), 3rd (2000-2007), 4th (2008-2015) and 5th (2016-2019). The CQI program was implemented between 2015 and 2016. The analysis included the total number of surgeries and the evolution of the most frequent procedures. A total of 105,599 CCVs were performed, with an annual mean of 2,964 procedures and mortality of 5,63%. When comparing the 4th and the 5th periods, the average global volume of surgeries was increased from 2,943 to 3,139 (p = 0.368), bypass graft (CABG), from 638 to 597 (p = 0.214), heart valve surgery, from 372 to 465 (p = 0.201), and congenital heart disease surgery, from 530 to 615 (p = 0.125). The average global mortality went from 7.8% to 5% (p < 0.0001); in CABG surgery, from 5.8% to 3.1% (p < 0.0001); in heart valve surgery, from 14% to 7.5% (p < 0.0001) and in congenital heart disease surgery, from 12.1% to 9.6% (p < 0.0001). In spite of a recent trend towards increased surgical volume, there was a significant decrease in operative mortality in the groups studied. After the implementation of the CQI program, the mortality rates were closer to international standards.


[Chronic heart failure: structural and functional deficit of our health policy?].
The problems concerning chronic heart failure can be summarized in 4 paradoxes which concern epidemiology, diagnosis, therapy, and financing respectively. Paradox I: The mortality due to chronic heart failure continues to increase worldwide despite a slow but significant decrease in mortality due to acute coronary syndromes. Paradox II: The clinical manifestations of chronic heart failure correlate poorly with the underlying pathophysiological progression. Paradox III: There is a striking discordance between the perception of evidence-based guidelines by the primary care physician and the actual reality in his/her private medical practice. Paradox IV: The inevitable increase in financial cost contrasts sharply with the many desperate attempts for cost reduction by the government. Heart Failure Clinic. Since the introduction of the first heart failure clinics in Sweden in 1990, numerous studies in various countries have emphasized the medical-cardiological and economical benefit of such organizations, mainly as a result of a substantial reduction of more than 40% in hospitalization. Moreover, a more central role is attributed to the primary care general practicioner.


Characteristics of dielectric-band modified single-cell photonic crystal lasers.
We demonstrate new types of dielectric-band photonic crystal lasers in a two-dimensional modified single-cell cavity with enlarged air holes. Finite-difference time-domain simulations performed in real and Fourier spaces show that the dielectric-band cavity modes originating from the first band edge point in the dielectric band have mode patterns that are distinguishable from conventional air-band cavity modes. In our experiment, the observed multimode lasing peaks are identified as the hexapole and the monopole dielectric-band cavity modes through the spectral positions and mode images. The thresholds of these lasers are measured as approximately 340 microW and approximately 450 microW, respectively, at room temperature. In addition, using the simulation based on the actual fabricated structures, quality factors and mode volumes are computed as 4900 and 1.09 (lambda/n)3 for the hexapole mode, and 4300 and 2.27 (lambda/n)3 for the monopole mode, respectively.


Autoinflammatory genes and susceptibility to psoriatic juvenile idiopathic arthritis.
To investigate the association of NLRP3, NOD2, MEFV, and PSTPIP1, genes that cause 4 of the autoinflammatory hereditary periodic fever syndromes (HPFS), with juvenile idiopathic arthritis (JIA). Fifty-one single-nucleotide polymorphisms (SNPs) across the 4 loci were investigated using MassArray genotyping in 950 Caucasian patients with JIA living in the UK and 728 ethnically matched healthy controls. Prior to Bonferroni correction for multiple testing, significant genotype associations between 6 SNPs in MEFV and JIA were observed and, in subgroup analysis, associations between 12 SNPs across all 4 loci and the subgroup of patients with psoriatic JIA were found. After Bonferroni correction for multiple testing, 2 genotype associations remained significant in the subgroup of patients with psoriatic JIA (MEFV SNP rs224204 [corrected P = 0.025] and NLRP3 SNP rs3806265 [corrected P = 0.04]). These findings support the use of monogenic loci as candidates for investigating the genetic component of complex disease and provide preliminary evidence of association between SNPs in autoinflammatory genes and psoriatic JIA. Our findings raise the interesting possibility of a shared disease mechanism between the HPFS and psoriatic JIA, potentially involving abnormal production of interleukin-1beta.


lontophoresis for Treatment of Shinsplints*.
It was hypothesized that shinsplints are the result of muscle inflammation induced by overuse; thus, as an inflammatory response, the application of an antiinflammatory medication to the involved muscles would eliminate the pain of shinsplints. The application of hydrocortisone by iontophoresis was effective in the elimination of shinsplint pain in 18 cases of shinsplints involving four male and eight female athletes. J Orthop Sports Phys Ther 1982;3(4):183-185.


Hydrogen bonding and C-H...X interactions in two triclinic phases of 4-carboxyquinolinium chloride monohydrate.
The title substance, C10H8NO2+.Cl-.H2O, crystallized in the centrosymmetric space group P1 in two phases, alpha and beta, each with one organic cation, one Cl- ion and one water molecule in the asymmetric unit. The principle structural difference in the two asymmetric units lies in the relative orientation of the water molecule. Three hydrogen bonds in the alpha phase have donor-acceptor distances (N...Cl or O...Cl) ranging from 3.052 (1) to 3.189 (2) A, while one has an O...O distance of 2.603 (3) A. Three hydrogen bonds in the beta phase have donor-acceptor distances (N...Cl or O...Cl) ranging from 3.044 (2) to 3.206 (2) A, while two have O...O distances of 2.586 (2) and 3.147 (3) A. The H atoms in all these hydrogen bonds are ordered. Each of these phases has five leading C-H...X interactions, for which the H...X distances are less than, or at most slightly greater than, the corresponding van der Waals radius sums. Taken together, these hydrogen bonds and C-H...X interactions form a three-dimensional network of interactions in each phase. The dihedral angle between the best-fit quinoline core plane and the carboxyl group plane is 25.9 (1) degree for the alpha phase and 20.0 (1) degree for the beta phase.


Neural correlates of metacognition: A critical perspective on current tasks.
Humans have a remarkable ability to reflect upon their behavior and mental processes, a capacity known as metacognition. Recent neurophysiological experiments have attempted to elucidate the neural correlates of metacognition in other species. Despite this increased attention, there is still no operational definition of metacognition and the ability of behavioral tasks to reflect metacognition is the subject of debate. The most widely used task for studying metacognition in animals, the uncertain-option task, has been criticized because it can be solved by simple associative mechanisms. Here we propose a broad perspective that generalizes those critiques to another task, post-decision wagering. Moreover, we extend this critical view to account for recent neurophysiological evidence. We argue these tasks are simple enough that any animal could solve them using very simple mechanisms such as sensory-motor associations. In this case, it is impossible to know whether all animals are metacognitive, or if the tasks are simply not appropriate. Therefore, we suggest using better defined concepts until a suitable task for metacognition is available.


The effects of in vivo antibiotics on neutrophil (PMN) activity in rabbits with peritonitis.
Antibiotics play an important role in helping the host fight infection; however, the direct cellular effect of antibiotics on polymorphonuclear cells remains undefined. Adherence, chemotaxis, phagocytosis, and superoxide anion production are important steps in the cascade of events initiated by the polymorphonucleocyte in bacterial killing. Previous studies have shown inhibition as well as stimulation of neutrophil antibacterial therapy by antibiotics. Peritoneal and blood polymorphonuclear neutrophils (PMN) respond differently to peritonitis and to external agents. The purpose of this study was to investigate the effects of in vivo clindamycin and netilmicin on infected rabbit peritoneal and blood polymorphonuclear adhesiveness, phagocytosis, chemotaxis, and superoxide anion production. Peritoneal and blood PMNs were obtained from rabbits which had undergone appendiceal devascularization 18 hr earlier: antibiotics were administered intramuscularly 1 hr prior to appendectomy and every 8 hr postoperatively for 5 days; these PMNs were compared to infected rabbits which did not receive antibiotics. Clindamycin and netilmicin in vivo cause significant inhibition of phagocytosis, peritoneal adhesiveness, and, when used in combination, blood adhesiveness and peritoneal superoxide anion production. No effects were seen on chemotaxis. Based on this data we conclude that antibiotics, while vitally important in fighting infections, may in and of themselves be agents of immunosuppression at the cellular level.


Rediscovery of an unusual chytridiaceous fungus new to the order Rhizophydiales.
The newest order in Chytridiomycota, Rhizophydiales, shows remarkable genetic divergence and zoospore ultrastructural diversity. From submersed mud from a marsh near an oligotrophic lake in the Patagonia region of Argentina, we isolated a previously undescribed chytrid in Rhizophydiales with unique 28S rRNA and ITS1-5.8S-ITS2 sequences and a distinctive zoospore and thallus. In a combined LSU + 5.8S phylogeny of representative taxa in the order, the nearest relative to this new species is Rhizophlyctis harderi, and their zoospores share several similar character states. A new genus is erected for this chytrid, the first to be brought into pure culture from this region.


Molecular basis for inner kinetochore configuration through RWD domain-peptide interactions.
Kinetochores are dynamic cellular structures that connect chromosomes to microtubules. They form from multi-protein assemblies that are evolutionarily conserved between yeasts and humans. One of these assemblies-COMA-consists of subunits Ame1CENP-U, Ctf19CENP-P, Mcm21CENP-O and Okp1CENP-Q A description of COMA molecular organization has so far been missing. We defined the subunit topology of COMA, bound with inner kinetochore proteins Nkp1 and Nkp2, from the yeast Kluyveromyces lactis, with nanoflow electrospray ionization mass spectrometry, and mapped intermolecular contacts with hydrogen-deuterium exchange coupled to mass spectrometry. Our data suggest that the essential Okp1 subunit is a multi-segmented nexus with distinct binding sites for Ame1, Nkp1-Nkp2 and Ctf19-Mcm21. Our crystal structure of the Ctf19-Mcm21 RWD domains bound with Okp1 shows the molecular contacts of this important inner kinetochore joint. The Ctf19-Mcm21 binding motif in Okp1 configures a branch of mitotic inner kinetochores, by tethering Ctf19-Mcm21 and Chl4CENP-N-Iml3CENP-L Absence of this motif results in dependence on the mitotic checkpoint for viability.


Inappropriate behaviours in a dental training environment: pilot of a UK-wide questionnaire.
Incivility and inappropriate behaviour in the workplace are topics of growing interest due to their impact on patient care and safety. Several surveys and campaigns have emerged highlighting the existence of a problem. However, the true scale is difficult to ascertain. The aim of this study is to determine the existence of inappropriate behaviours within the UK dental training environment.An anonymous pilot questionnaire was distributed across multiple platforms reaching out to dental professionals within training environments, inviting responses between July 2022 and October 2022. A total of 215 responses were received. The vast majority (73.2%) felt that inappropriate behaviour is a problem within UK dental training. Senior colleagues were identified as perpetrators in 88% of responses. Most respondents (66%) reported feeling uncomfortable raising the issue, and when raised, 30% felt unsupported. Only 9% felt confident that action was taken after the issue was reported. Belittling was experienced and witnessed most commonly.The feedback received reveals the existence of inappropriate behaviours within dental training environments. Qualitative feedback indicates that if left unaddressed, the impact of such behaviour may persist long-term. Further research is required to address this issue, improve dental training conditions and job satisfaction.


Integrated and High-Throughput Approach for Sensitive Analysis of Tyrosine Phosphoproteome.
Tyrosine phosphorylation (pTyr) regulates various signaling pathways under normal and cancerous states. Due to their low abundance and transient and dynamic natures, systematic profiling of pTyr sites is challenging. Antibody and engineered binding domain-based approaches have been well applied to pTyr peptide enrichment. However, traditional methods have the disadvantage of a long sample preparation process, which makes them unsuitable for processing limited amount of samples, especially in a high-throughput manner. In this study we developed a 96-well microplate-based approach to integrate all the sample preparation steps starting from cell culture to MS-compatible pTyr peptide enrichment in three consecutive 96-well microplates. By assembling an engineered SH2 domain onto a microplate, nonspecific adsorption of phosphopeptides is greatly reduced, which allows us to remove the Ti-IMAC purification and three C18 desalting steps (after digestion, pTyr enrichment, and Ti-IMAC purification) and, therefore, greatly simplifies the entire pTyr peptide enrichment workflow, especially when processing a large number of samples. Starting with 96-well microplate-cultured, pervanadate-stimulated cells, our approach could enrich 21% more pTyr sites than the traditional serial pTyr enrichment approach and showed good sensitivity and reproducibility in the range of 200 ng to 200 μg peptides. Importantly, we applied this approach to profile tyrosine kinase inhibitor-mediated EGFR signaling pathway and could well differentiate the distinct response of different pTyr sites. Collectively, the integrated 96-well microplate-based approach is valuable for profiling pTyr sites from limited biological samples and in a high-throughput manner.


Self-assembly of colloidal one-dimensional nanocrystals.
The ability of nanoscopic materials to self-organize into large-scale assembly structures that exhibit unique collective properties has opened up new and exciting opportunities in the field of nanotechnology. Although earlier work on nanoscale self-assembly has focused on colloidal spherical nanocrystals as building blocks, there has been significant interest in recent years in the self-assembly of colloidal nanocrystals having well-defined facets or anisotropic shapes. In this review, particular attention is drawn to anisotropic one-dimensional (1D) nanocrystals, notably nanorods and nanowires, which can be arranged into a multitude of higher-order assembly structures. Different strategies have been developed to realize self-assembly of colloidal 1D nanocrystals and these are highlighted in the first part of this review. Self-assembly can take place (1) on substrates through evaporation control, external field facilitation and template use; (2) at interfaces, such as the liquid-liquid and the gas-liquid interface; and (3) in solutions via chemical bonding, depletion attraction forces and linker-mediated interactions. The choice of a self-assembly approach is pivotal to achieving the desired assembly configuration with properties that can be exploited for functional device applications. In the subsequent sections, the various assembly structures that have been created through 1D nanocrystal self-assembly are presented. These organized structures are broadly categorized into non-close-packed and close-packed configurations, and are further classified based on the different types of 1D nanocrystal alignment (side-by-side and end-to-end), orientation (horizontal and vertical) and ordering (nematic and smectic), and depending on the dimensionality of the structure (2D and 3D). The conditions under which different types of arrangements are achieved are also discussed.


Thyroid hormones act primarily within the brain to promote the seasonal inhibition of luteinizing hormone secretion in the ewe.
In the ewe, thyroid hormones are required for the seasonal suppression of GnRH and LH secretion, thereby maintaining an annual rhythm in reproductive activity. The primary site of action of thyroid hormones is unknown; in particular, there is no evidence to distinguish a central from a peripheral action. In this study, we test the hypothesis that thyroid hormones can act directly within the brain to promote GnRH/LH seasonal inhibition. Ovariectomized estradiol-treated ewes were thyroidectomized late in the breeding season to prevent seasonal LH inhibition. T4 was then infused for 3 months, either peripherally or centrally. Neuroendocrine reproductive state was monitored by assaying the LH concentration in biweekly blood samples. Central infusion of low dose T4, which restored a physiological concentration of the hormone in cerebrospinal fluid of these thyroidectomized ewes, promoted the neuroendocrine changes that lead to anestrus. The serum LH concentration in these animals fell at the same time as the seasonal LH decline in euthyroid controls. Neither this same T4 dose infused peripherally nor vehicle infused centrally was effective; LH remained elevated, signifying blockade of the mechanism for anestrus. Our results provide strong evidence that thyroid hormones can act directly within the brain to promote seasonal inhibition of neuroendocrine reproductive function in the ewe.


Dietary fluoride intake from supplements and communal water supplies.
Artificial fluoridation of communal water supplies has resulted in 50% to 60% reductions in dental caries experience accompanied by very little fluorosis. Although vitamin-mineral fluoride supplements also have resulted in significant caries reductions, data are not readily available in the literature in regard to enamel fluorosis in the permanent dentition of children who have received supplements in early infancy. In comparison to estimates of fluoride intake from communal water supplies, the recommended dosage from fluoride supplements results in daily doses that may be two to six times greater, on the average, for the infant less than age 6 months and three times greater for the average 3-year-old. Estimates from both sources bear careful consideration in light of available knowledge and are not necessarily indications for a revision in presently acceptable practice.


Gene-gene interactions among coding genes of iron-homeostasis proteins and APOE-alleles in cognitive impairment diseases.
Cognitive impairments of different aetiology share alterations in iron and lipid homeostasis with mutual relationships. Since iron and cholesterol accumulation impact on neurodegenerative disease, the associated gene variants are appealing candidate targets for risk and disease progression assessment. In this light, we explored the role of common single nucleotide polymorphisms (SNPs) in the main iron homeostasis genes and in the main lipoprotein transporter gene (APOE) in a cohort of 765 patients with dementia of different origin: Alzheimer's disease (AD) n = 276; vascular dementia (VaD), n = 255; mild cognitive impairment (MCI), n = 234; and in normal controls (n = 1086). In details, four genes of iron homeostasis (Hemochromatosis (HFE: C282Y, H63D), Ferroportin (FPN1: -8CG), Hepcidin (HAMP: -582AG), Transferrin (TF: P570S)), and the three major alleles of APOE (APOE2, APOE3, APOE4) were analyzed to explore causative interactions and synergies. In single analysis, HFE 282Y allele yielded a 3-fold risk reduction in the whole cohort of patients (P<0.0001), confirmed in AD and VaD, reaching a 5-fold risk reduction in MCI (P = 0.0019). The other iron SNPs slightly associated with risk reduction whereas APOE4 allele resulted in increased risk, reaching more than 7-fold increased risk in AD homozygotes (P = 0.001), confirmed to a lower extent in VaD and MCI (P = 0.038 and P = 0.013 respectively) as well as in the whole group (P<0.0001). Comparisons of Mini Mental State Examination (MMSE) among AD showed appreciable lowering in APOE4 carriers (P = 0.038), confirmed in the whole cohort of patients (P = 0.018). In interaction analysis, the HFE 282Y allele completely extinguished the APOE4 allele associated risk. Conversely, the coexistence in patients of a substantial number of iron SNPs accrued the APOE4 detrimental effect on MMSE. Overall, the analysis highlighted how a specific iron-allele burden, defined as different combinations of iron gene variants, might have different effects on cognitive impairment and might modulate the effects of established genetic risk factors such as APOE4. Our results suggest that established genetic risk factors might be affected by specific genetic backgrounds, making patients differently suited to manage iron accumulation adding new genetic insights in neurodegeneration. The recently recognized interconnections between iron and lipids, suggest that these pathways might share more than expected. We therefore extended to additional iron gene variants the newly proposed influencing mechanisms that HFE gene has on cholesterol metabolism. Our results have a strong translational potential promoting new pharmacogenetics studies on therapeutic target identification aimed at optimally tuning brain iron levels.


Control of blood vessel structure: insights from theoretical models.
Blood vessels are capable of continuous structural adaptation in response to changing local conditions and functional requirements. Theoretical modeling approaches have stimulated the development of new concepts in this area and have allowed investigation of the complex relations between adaptive responses to multiple stimuli and resulting functional properties of vascular networks. Early analyses based on a minimum-work principle predicted uniform wall shear stress in all segments of vascular networks and led to the concept that vessel diameter is controlled by a feedback system based on responses to wall shear stress. Vascular reactions to changes in transmural pressure suggested feedback control of circumferential wall stress. However, theoretical simulations of network adaptation showed that these two mechanisms cannot, by themselves, lead to stable and realistic network structures. Models combining reactions to fluid shear stress, circumferential stress, and metabolic status of tissue, with propagation of stimuli upstream and downstream along vascular segments, are needed to explain stable and functionally adequate adaptation of vascular structure. Such models provide a basis for predicting the response of vascular segments exposed to altered conditions, as, for example, in vascular grafts.


International consensus for a dissection room quality system (DRQS): A Delphi panel study.
Dissection Rooms (DRs) are key facilities that allow teaching and research on human anatomy, where students and researchers work with human bodies to acquire, increase, or create new knowledge. Usually, DRs work with a Body Donation Program (BDP), where living donors bequeath their bodies for use in teaching and research after they expire. Despite DRs being part of universities worldwide, no common guidelines, regulations, or quality management systems (QMS) exist that could be applied to different countries. With that purpose in mind, we aimed to develop a QMS that could be applied to DRs globally, using a Delphi panel to achieve consensus about the items that should constitute the QMS. The panel was constituted by 20 anatomy professors from 20 different countries, and the 167 standards to create the rules or guidelines that constitute the QMS were divided in five categories: direction, body donation, students, instructors, and research. After two rounds of revisions, 150 standards were considered "essential" or "important" by more than 70% of the participants, thus being incorporated to the Dissection Room Quality System (DRQS). The results of this panel represent a minimum list of items of the DRQS for improving the functioning of DRs globally.


Occupational health problems among tobacco processing workers: a preliminary study.
Occupational health problems among 100 tobacco processing workers were investigated. Symptoms, including vomiting, giddiness, headache, etc. were found among 69 exposed subjects. It was also observed that the excretion rate of nicotine and cotinine increased among exposed subjects. Biochemical parameters were found to be within the normal range. Electrocardiographic findings were non-specific and clinically there was no evidence of hypertension or ischemic heart disease. Therefore, the symptoms in tobacco processing workers might possibly result from mild nicotine toxicity.


Establishing genetic manipulation for novel strains of human gut bacteria.
Recent years have seen the development of high-accuracy and high-throughput genetic manipulation techniques, which have greatly improved our understanding of genetically tractable microbes. However, challenges remain in establishing genetic manipulation techniques in novel organisms, owing largely to exogenous DNA defence mechanisms, lack of selectable markers, lack of efficient methods to introduce exogenous DNA and an inability of genetic vectors to replicate in their new host. In this review, we describe some of the techniques that are available for genetic manipulation of novel microorganisms. While many reviews exist that focus on the final step in genetic manipulation, the editing of recipient DNA, we particularly focus on the first step in this process, the transfer of exogenous DNA into a strain of interest. Examples illustrating the use of these techniques are provided for a selection of human gut bacteria in which genetic tractability has been established, such as Bifidobacterium, Bacteroides and Roseburia. Ultimately, this review aims to provide an information source for researchers interested in developing genetic manipulation techniques for novel bacterial strains, particularly those of the human gut microbiota.


Soft tissue osteochondroma: case report and immunohistochemistry for parathyroid hormone-related protein.
Surface lesions of bone usually present little diagnostic dilemma because the majority are conventional osteochondromas. Other surface bone lesions include periosteal chondroma, periosteal chondrosarcoma, and parosteal osteosarcoma. Mineralized soft tissue lesions such as myositis ossificans, synovial chondroma, and synovial sarcoma may present in a similar fashion when they occur in a juxtaarticular position. The soft tissue osteochondroma or paraarticular osteochondroma may simulate some of these more aggressive tumors, and its recognition is important to avoid overtreatment. A case of an 11-year-old male with a soft tissue osteochondroma is reported to illustrate the characteristic radiographic and histological features of this rare entity. No prior reports have examined soft tissue osteochondroma for expression of parathyroid hormone related protein, an established cartilage tumor proliferative mitogen.


Mixture effects of a fungicide and an antibiotic: Assessment and prediction using a decomposer-detritivore system.
Antimicrobials, such as fungicides and antibiotics, pose a risk for microbial decomposers (i.e., bacteria and aquatic fungi) and invertebrate detritivores (i.e., shredders) that play a pivotal role in the ecosystem function of leaf litter breakdown. Although waterborne toxicity and diet-related effects (i.e., dietary exposure and microorganism-mediated alterations in food quality for shredders) of fungicides and antibiotics on decomposer-detritivore systems have been increasingly documented, their joint effect is unknown. We therefore assessed waterborne and dietary effects of an antimicrobial mixture consisting of the fungicide azoxystrobin (AZO) and the antibiotic ciprofloxacin (CIP) on microbial decomposers and the shredder Gammarus fossarum using a tiered approach. We compared effect sizes measured in the present study with model predictions (i.e., independent action) based on published data. During a 7-day feeding activity assay quantifying waterborne toxicity in G. fossarum, the leaf consumption of gammarids was reduced by ∼60 % compared to the control when subjected to the mixture at concentrations of each component causing a 20 % reduction in the same response variable when applied individually. Moreover, the selective feeding of gammarids during the food choice assay indicated alterations in food quality induced by the antimicrobial mixture. The food selection and, in addition, the decrease in microbial leaf decomposition is likely linked to changes in leaf-associated bacteria and fungi. During a long-term assay, energy processing, growth and energy reserves of gammarids were increased in presence of 15 and 500 μg/L of AZO and CIP, respectively, through the dietary pathway. These physiological responses were probably driven by CIP-induced alterations in the gut microbiome or immune system of gammarids. In general, model predictions matched observed effects caused by waterborne exposure on the leaf consumption, energy processing and growth of gammarids during short- and long-term assays, respectively. However, when complex horizontal (bacteria and aquatic fungi) and vertical (leaf-associated microorganisms and shredders) interactions were involved, model predictions partly over- or underestimated mixture effects. Therefore, the present study identifies uncertainties of mixture effect predictions for complex biological systems calling for studies targeting the underlying processes and mechanisms.


Effects of a novel direct-fed microbial on growth performance, carcass characteristics, nutrient digestibility, and ruminal morphology of beef feedlot steers.
The effects of a novel direct-fed microbial on feedlot performance, carcass characteristics, digestibility, ruminal morphology, and volatile fatty acid (VFA) profile of finishing steers were evaluated. Single source Angus-crossbred yearling steers (n = 144; initial BW = 371 ± 19 kg) were used in a randomized complete block design. Steers were blocked by initial BW and randomly assigned to treatments (12 pens/treatment; 4 steers/pen). Treatments included: A) CONTROL (no DFM, tylosin, or monensin; B) MONTY (monensin sodium [330 mg/animal-daily] and tylosin phosphate [90 mg/animal-daily]); and C) MONPRO (monensin sodium [same as previous] and L. salivarius L28 [1 × 10⁶ CFU/animal-daily]). Treatments were included in a steam-flaked corn-based finisher diet offered once daily using a clean-bunk management for approximately 149 d. The digestibility assessment was performed from d 70 to d 74. Ruminal fluid and rumen tissue samples were collected at the slaughter for VFA profile and papillae morphology analyses, respectively. Data were analyzed using the GLIMMIX procedure of SAS with pen serving as the experimental unit, treatment as fixed effect, and BW block as random effect. Steers offered MONPRO had on average 5.3 % less (P < 0.01) dry matter intake (9.56 kg/d) compared to either CONTROL (10.16 kg/d) or MONTY (9.96 kg/d). The carcass-adjusted final BW (613 kg; P = 0.23), overall ADG (1.64 kg/d; P = 0.23), and gain-efficiency (0.165; P = 0.61) were not affected by treatments. Steers offered CONTROL had greater (P < 0.01) marbling score and tended (P = 0.06) to have less carcasses grading Select and tended (P = 0.10) to have more carcasses grading Upper-Choice, while other carcass characteristics and liver-abscesses were not affected (P ≥ 0.23) by treatments. The digestibility of nutrients (P ≥ 0.13) and the ruminal VFA profile (P ≥ 0.12) were not affected by treatments. Steers offered MONPRO tended (P = 0.09) to have 16% greater average papillae number compared to other treatments. Yearlings offered finishing diets containing L. salivarius L28 plus monensin did not affect growth performance, digestibility, or ruminal VFA, but reduced feed intake. Carcass quality was negatively affected by treatments, while animals consuming L. salivarius L28 and monensin tended to improve ruminal morphology. Current findings in ruminal morphology and feed intake may warrant further assessment of diets containing L. salivarius L28 on beef cattle food safety aspects.


Amanita mushroom poisoning: efficacy of aggressive treatment of two dogs.
Amatoxins, the primary toxins found in mushrooms of the genus Amanita, are very toxic to dogs. Acute fulminant liver failure and death can occur within a few days of ingestion. By their curious nature, dogs, especially young dogs, are prone to ingest mushrooms. Early identification of suspect mushrooms, and prompt emergency measures aimed at decreasing absorption of the toxins can improve the chance of survival. Knowing the major clinical syndromes associated with Amanita mushroom toxicosis can help direct the treatment and supportive care of affected animals and improve survival rates. We describe 2 cases in dogs with confirmed ingestion of Amanita phalloides and Amanita ocreata resulting in fulminant liver failure. Death occurred in 1 dog despite aggressive treatment measures including hemoperfusion, while aggressive measures resulted in a favorable outcome in the other dog.


Analysis of Automotive Paint Smears Using Attenuated Total Reflection Infrared Microscopy.
Paint smears represent a type of automotive paint sample found at a crime scene that is problematic for forensic automotive paint examiners to analyze as there are no reference materials present in automotive paint databases to generate hit-lists of potential suspect vehicles. Realistic paint smears are difficult to create in a laboratory and have also proven challenging to analyze because of the mixing of the various automotive paint layers. A procedure based on an impact tester has been developed to create smears to simulate paint transfer between vehicles during a collision. Data collected from 24 original equipment manufacturer (OEM) paints in simulated collisions using an impact tester with a steel (inert) substrate to simulate vehicle to vehicle collisions shows that attenuated total reflection infrared microscopy can isolate individual paint layers. For each OEM paint sample, the corresponding smear obtained was dependent upon the conditions used. By varying these conditions, the number of distinct layers obtained could be tuned for each of the OEM paints investigated. Furthermore, the IR spectrum of each layer extracted from the paint smear using alternating least squares was found to compare favorably to an in-house OEM paint infrared spectral library for each layer as the correct match (make and model of the vehicle from which the smear originated) was always found as a top five hit in the hit-list. The results of this study indicate that paint smears developed using an impactor can serve as the basis of realistic proficiency tests for forensic laboratories.


A simple method for rapid identification of Sphaerophorus necrophorus isolates.
A hemagglutination inhibition test for the rapid identification of Sphaerophorus necrophorus is described. Erythrocytes from six species of animals were tested and human cells were found to be the best agglutination indicators. Antiserum prepared in rabbits was found to be specific for S. necrophorus hemagglutinins when tested against 20 isolates of S. necrophorus and 117 other bacteria belonging to 22 genera. The possibility of using a hemagglutination inhibition test for the detection of bovine necrobacillosis was explored.


Strength and body weight in US children and adolescents.
Regular aerobic and muscle-strengthening physical activity in youth has been positively associated with health and may help prevent obesity. The purpose of this study is to provide reference values on 4 core, upper, and lower body measures of muscle strength among US children and adolescents and to investigate the association between these measures of strength and weight status. We assessed muscular strength using 4 different tests (plank, modified pull-up, knee extension, and grip strength) in 1224 youth aged 6 to 15 years collected during the 2012 National Health and Nutrition Examination Survey National Youth Fitness Survey. Mean and median estimates are provided by gender, age, and weight status. Weight status was defined based on standard categories of obesity, overweight, normal weight, and underweight using the gender-specific BMI-for-age Centers for Disease Control and Prevention growth charts. There were significant positive trends with age for each of the strength tests (P < .001) except the modified pull-up among girls. The length of time the plank was held decreased as weight status increased for both girls and boys (P < .001). As weight status increased the number of modified pull-ups decreased (P < .001 boys and girls). Scores on the knee extension increased as weight status increased (P < .01). Grip strength increased as weight status increased (P < .01). Increasing weight status had a negative association with measures of strength that involved lifting the body, but was associated with improved performances on tests that did not involve lifting the body.


Cocaine-like effects of intravenous procaine in cocaine addicts.
Pharmacological treatments that alter dopaminergic functioning have not lessened cocaine use in addicted patients. Non-dopaminergic mechanisms may therefore be important in the chronic use of cocaine. Procaine, like cocaine, is a local anesthetic, but has only 1% of cocaine's affinity for the dopamine reuptake receptor. In order to assess the subjective effects of procaine and its similarity to cocaine, we administered procaine to nine cocaine-dependent subjects. Patients 2-3 weeks abstinent were administered placebo, low dose procaine (0.46 mg/kg), and high dose procaine (1.84 mg/kg procaine) over a single 2-hour session. Patients were assessed for craving and similarity to cocaine experience and were administered the Symptom Checklist 90 Revised (SCL90R). High dose procaine was identified as similar to cocaine and induced significant cocaine craving. High dose procaine also induced significant elevations in somatization, obsessive-compulsive symptoms, phobic anxiety, interpersonal sensitivity, anxiety, positive symptoms and global severity (from the SCL90R). Our findings suggest that procaine shares subjective effects similar to cocaine, despite a much lower affinity for the dopamine reuptake receptor. Procaine may be a useful tool to explore non-dopaminergic mechanisms of cocaine's reinforcing and addictive properties.


High-Resolution Imaging of Interaction Between Thrombus and Stent-Retriever in Patients With Acute Ischemic Stroke.
Currently, acute ischemic stroke is still a leading cause of mortality and morbidity. Approximately 2 years ago, mechanical thrombectomy was proven beneficial as a revolutionary new therapy for stroke in the MR-CLEAN trial (A Multicenter Randomized Clinical Trial of Endovascular Treatment for Acute Ischemic Stroke in the Netherlands). However, the mechanisms by which the thrombectomy device, or stent-retriever, interacts with the thrombus are largely unknown. A better understanding could lead to improved efficacy of mechanical thrombectomy devices. Seven stent-retrievers with thrombi still entrapped were collected directly after thrombectomy. The stent-retrievers were studied using micro computed tomography, followed by scanning electron microscopy and light microscopy. Two independent observers rated interaction type and thrombus surface structure (porous filamentous or dense) at the interaction sites.A total of 79 interaction sites between thrombus and stent-retriever were categorized. Thrombus-stent-retriever interaction was found to be adhesive (n=44; 56%) or mechanical (n=35; 44%). Adhesive interaction was most frequently observed at interaction sites with a dense surface, compared with interaction sites with a porous filamentous fibrin surface (38/58; 66% versus 6/21; 29%, P=0.011). The interaction between thrombus and stent-retriever was predominantly adhesive, not mechanical. Adhesive interaction was strongly associated with the presence of a dense thrombus surface without a porous filamentous fibrin network.


[Knowledge on chronic oral anticoagulation of patients followed up in a specialized outpatient clinic].
This study was carried out to assess patients' knowledge on anticoagulant therapy in an outpatient clinic. A questionnaire with ten questions with closed answers was applied. Correct answers to about 80% of the questions were considered satisfactory. Out of the 120 evaluated patients, 77 (64.1%) showed satisfactory knowledge, with an average of 7.8 +/- 1.7 correct answers. As to the International Normalized Ratio (INR), 69.4% of the sample was beyond therapeutic range, and 42.5% had hemorrhages in the last 6 months. Factors such as education level, income, hemorrhagic event, anticoagulation time, and gender did not show statistically significant difference. There is an obvious need for a systematic follow-up by health care professionals, using educational measures, to detect the difficulties faced by patients, maintaining an optimal INR.


Models for the analysis of repeated continuous outcome measures in clinical trials.
Repeated continuous outcome measures are common in clinical trials. In this tutorial style paper, using data collected from a trial evaluating an intervention for managing asthma and chronic obstructive pulmonary disease, we demonstrate ways of statistically analysing such data to answer frequently encountered clinical research questions. We illustrate the use of linear mixed effects modelling in doing so and discuss its advantages over several other commonly used approaches. The methods described in this paper can easily be carried out using standard statistical software.


Effects of ultrasonic noise on the human body-a bibliographic review.
Industrial noise in the working environment has adverse effects on human hearing; literature and private studies confirm that. It has been determined that significant changes in the hearing threshold level occur in the high frequency audiometry, i.e., in the 8-20 kHz frequency range. Therefore, it is important to determine the effect of ultrasonic noise (10-40 kHz) on the human body in the working environment. This review describes hearing and nonhearing effects (thermal effects, subjective symptoms and functional changes) of the exposure to noise emitted by ultrasound devices. Many countries have standard health exposure limits to prevent effects of the exposure to ultrasonic noise in the working environment.


Assigning ICD-O-3 Codes to Pathology Reports using Neural Multi-Task Training with Hierarchical Regularization.
Tracking population-level cancer information is essential for researchers, clinicians, policymakers, and the public. Unfortunately, much of the information is stored as unstructured data in pathology reports. Thus, too process the information, we require either automated extraction techniques or manual curation. Moreover, many of the cancer-related concepts appear infrequently in real-world training datasets. Automated extraction is difficult because of the limited data. This study introduces a novel technique that incorporates structured expert knowledge to improve histology and topography code classification models. Using pathology reports collected from the Kentucky Cancer Registry, we introduce a novel multi-task training approach with hierarchical regularization that incorporates structured information about the International Classification of Diseases for Oncology, 3rd Edition classes to improve predictive performance. Overall, we find that our method improves both micro and macro F1. For macro F1, we achieve up to a 6% absolute improvement for topography codes and up to 4% absolute improvement for histology codes.


Interstitial cells of Cajal: update on basic and clinical science.
The basic science and clinical interest in the networks of interstitial cells of Cajal (ICC) keep growing, and here, research from 2010 to mid-2013 is highlighted. High-resolution gastrointestinal manometry and spatiotemporal mapping are bringing exciting new insights into motor patterns, their function and their myogenic and neurogenic origins, as well as the role of ICC. Critically important knowledge is emerging on the partaking of PDGFRα+ cells in ICC pacemaker networks. Evidence is emerging that ICC and PDGFRα+ cells have unique direct roles in muscle innervation. Chronic constipation is associated with loss and injury to ICC, which is stimulating extensive research into maintenance and repair of ICC after injury. In gastroparesis, high-resolution electrical and mechanical studies are beginning to elucidate the pathophysiological role of ICC and the pacemaker system in this condition. Receptors and ion channels that play a role in ICC function are being discovered and characterized, which paves the way for pharmacological interventions in gut motility disorders through ICC.


Insulin increases endothelin-1-evoked intracellular free calcium responses by increased ET(A) receptor expression in rat aortic smooth muscle cells.
While insulin is known to promote vascular smooth muscle (VSM) relaxation, it also enhances endothelin-1 (ET-1) secretion and action in conditions such as NIDDM and hypertension. We examined the effect of insulin pretreatment on intracellular free calcium ([Ca2+]i) responses to ET-1 in cultured aortic smooth muscle cells (ASMCs) isolated from Sprague-Dawley (SD) rats and measured ET(A) receptor characteristics and ET-1-evoked tension responses in aorta obtained from insulin-resistant, hyperinsulinemic Zucker-obese (ZO) and control Zucker-lean (ZL) rats. Pretreatment of rat ASMCs with insulin (10 nmol/l for 24 h) failed to affect basal [Ca2+]i levels but led to a significant increase in peak [Ca2+]i response (1.7-fold; P < 0.01) to ET-1. The responses to IRL-1620 (an ET(B) selective agonist), ANG II, and vasopressin remained unaffected. ET-1-evoked peak [Ca2+]i responses were significantly attenuated by the inclusion of the ET(A) antagonist, BQ123, in both groups. The ET(B) antagonist, BQ788, abolished [Ca2+]i responses to IRL-1620 but failed to affect the exaggerated [Ca2+]i responses to ET-1. Saturation binding studies revealed a twofold increase (P < 0.01) in maximal number of binding sites labeled by 125I-labeled ET-1 in insulin-pretreated cells and no significant differences in sites labeled by 125I-labeled IRL-1620 between control and treatment groups. Northern blot analysis revealed an increase in ET(A) mRNA levels after insulin pretreatment for 20 h, an effect that was blocked by genistein, actinomycin D, and cycloheximide. Maximal tension development to ET-1 was significantly greater (P < 0.01), and microsomal binding studies using [3H]BQ-123 revealed a twofold higher number of ET(A) specific binding sites (P < 0.01) in aorta from ZO rats compared with that of ZL rats. These data suggest that insulin exaggerates ET-1-evoked peak [Ca2+]i responses via increased vascular ET(A) receptor expression, which may contribute to enhanced vasoconstriction observed in hyperinsulinemic states.


A Q-TWiST analysis comparing panitumumab plus best supportive care (BSC) with BSC alone in patients with wild-type KRAS metastatic colorectal cancer.
Panitumumab+best supportive care (BSC) significantly improved progression-free survival (PFS) vs BSC alone in patients with chemo-refractory wild-type KRAS metastatic colorectal cancer (mCRC). We applied the quality-adjusted time without symptoms of disease or toxicity (Q-TWiST) analysis to provide an integrated measure of clinical benefit, with the objective of comparing quality-adjusted survival between the two arms. As the trial design allowed patients on BSC alone to receive panitumumab after disease progression, which confounded overall survival (OS), the focus of this analysis was on PFS. For each treatment group, the time spent in the toxicity (grade 3 or 4 adverse events; TOX), time without symptoms of disease or toxicity (TWiST), and relapse (after disease progression; REL) states were estimated by the product-limit method, and adjusted using utility weights derived from patient-reported EuroQoL 5-dimensions measures. Sensitivity analyses were performed in which utility weights (varying from 0 to 1) were applied to time in the TOX and REL health states. There was a significant difference between groups favouring panitumumab+BSC in quality-adjusted PFS (12.3 weeks vs 5.8 weeks, respectively, P<0.0001) and quality-adjusted OS (P=0.0303). In patients with chemo-refractory wild-type KRAS mCRC, panitumumab+BSC significantly improved quality-adjusted survival compared with BSC alone.


Effect of X-irradiation on adenylate cyclase activity and cyclic AMP content of primary human fibroblasts.
Ionizing radiation provokes an increase of the cAMP level in several organs and body fluids. After reviewing the relevant literature we present the results of our own experiments on primary human fibroblasts. X-irradiation at doses of 0.5 and 2.5 Gy in vitro evoked a rapid and reversible increase of adenylate cyclase enzyme activity. A significant increase in cAMP level of these cells was also observed. Adenylate cyclase was usually localized basolaterally on the surface of unirradiated cells, while irradiation resulted in a modification of distribution, i.e., the enzyme activity also appeared in apical localization.


Integrative Genomic Analysis Identifies the Core Transcriptional Hallmarks of Human Hepatocellular Carcinoma.
Integrative genomics helped characterize molecular heterogeneity in hepatocellular carcinoma (HCC), leading to targeted drug candidates for specific HCC subtypes. However, no consensus was achieved for genes and pathways commonly altered in HCC. Here, we performed a meta-analysis of 15 independent datasets (n = 784 human HCC) and identified a comprehensive signature consisting of 935 genes commonly deregulated in HCC as compared with the surrounding nontumor tissue. In the HCC signature, upregulated genes were linked to early genomic alterations in hepatocarcinogenesis, particularly gains of 1q and 8q. The HCC signature covered well-established cancer hallmarks, such as proliferation, metabolic reprogramming, and microenvironment remodeling, together with specific hallmarks associated with protein turnover and epigenetics. Subsequently, the HCC signature enabled us to assess the efficacy of signature-relevant drug candidates, including histone deacetylase inhibitors that specifically reduced the viability of six human HCC cell lines. Overall, this integrative genomics approach identified cancer hallmarks recurrently altered in human HCC that may be targeted by specific drugs. Combined therapies targeting common and subtype-specific cancer networks may represent a relevant therapeutic strategy in liver cancer. Cancer Res; 76(21); 6374-81. ©2016 AACR.


Gallic acid exerts a protective or an anti-proliferative effect on glioma T98G cells via dose-dependent epigenetic regulation mediated by miRNAs.
Glioblastoma multiforme (GBM) is the most malignant primary brain tumor in adulthood, characterized by very high recurrence. Following the limited results for conventional therapies, novel therapeutic agents are under investigation. Among the putative new molecules, gallic acid (GA) represents a promising new anticancer drug. The anticancer effect of this drug has been based on its antioxidant effects. The aim of the present study was to investigate the toxic effects of GA on the T98G human glioblastoma cell line and its capacity to modulate the expression of microRNAs targeting the genes involved in tumor growth and invasion. Cytotoxicity, clonogenic ability and cell migration after GA treatment were tested. Moreover, the expression of miRNAs that target genes for antioxidant mitochondrial enzymes (mir-17-3p), p-21 protein (mir-21-5p) and ATM (mir-421-5p) was determined by qRT-PCR. The results confirmed in the T98G cells the anti-proliferative effect of GA reported for other glioma cell lines and showed that the miRNA expression changes depending on GA concentrations. Different GA concentrations can determine a protective or a toxic effect on tumor cells. Thus, the key for GA to induce a specific anticancer action is to use an optimal concentration that avoids these twin effects.


Enhanced Colorimetric Differentiation between Staphylococcus aureus and Pseudomonas aeruginosa Using a Shape-Encoded Sensor Hydrogel.
Herein, we demonstrate a combined fluorescent probe/shape-encoded hydrogel strategy for the fast, sensitive, and selective detection of bacterial species via their characteristic enzymes. A poly(vinyl alcohol) (PVA) hydrogel loaded with the fluorescent probe N,N'-(3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-3',6'-diyl)bis(2,2,3,3,3-pentafluoropropanamide) (ACS-HNE) was designed for the detection of elastase, an enzyme produced by Pseudomonas aeruginosa. Likewise, a chitosan-derived hydrogel was loaded with the fluorescent probe 4-methylumbelliferyl-α-d-glucopyranoside (MUD) by entrapment for the selective detection of α-glucosidase, an enzyme produced by Staphylococcus aureus. For an observation time of 60 min, limits of detection (LODs) of ≤20 nM for elastase and ≤30 pM for α-glucosidase were obtained, which in the latter case is 3 orders of magnitude better than related chitosan systems with covalently coupled substrate. To illustrate the potential utility of these highly sensitive sensor hydrogels as a simple point-of-care test system, shaped hydrogel slabs representing the letters P and S were manufactured to detect P. aeruginosa and S. aureus, respectively. These shapes were shown to provide an additional unique color code under UV illumination corresponding to the characteristic enzyme produced by the corresponding bacteria. This study shows potential for the future development of an effective and simple point-of-care test for the rapid identification of bacterial species that can be operated by nonspecialists.


Kinetics of dodecanedioic acid triglyceride in rats.
The kinetics of the triglyceride of dodecanedioic acid (TGDA) has been investigated in 30 male Wistar rats after a rapid intravenous bolus injection. TGDA and its product of hydrolysis, nonesterified dodecanedioic acid (NEDA), were measured in plasma samples taken at different times using an improved high-performance liquid chromatographic method. The 24-h urinary excretion of TGDA was 1.54 +/- 0.37 micromol, corresponding to approximately 0.67% of the administered amount. Several kinetics models were considered, including central and peripheral compartments for the triglyceride and the free forms and expressing transports between compartments with combinations of linear, carrier-limited, or time-varying mechanisms. The parameter estimates of the kinetics of TGDA and of NEDA were finally obtained using a three-compartment model in which the transfer of TGDA to NEDA was assumed to be linear, through a peripheral compartment, and the tissue uptake of NEDA was assumed to be carrier limited. TGDA had a large volume of distribution ( approximately 0.5 l/kg body wt) with a fast disappearance rate from plasma (0.42 min-1), whereas NEDA had a very small volume of distribution ( approximately 0.04 l/kg body wt) and a tissue uptake with maximal transport rate of 0.636 mM/min. In conclusion, this first study on the triglyceride form of dodecanedioic acid indicates that it is rapidly hydrolyzed and that both triglyceride and nonesterified forms are excreted in the urine to a very low extent. The tissue uptake rate of NEDA is consistent with the possibility of achieving substantial energy delivery, should it be added to parenteral nutrition formulations. Furthermore, the amount of sodium administered with the triglyceride form is one-half of that necessary with the free diacid.


The minus end-directed motor Kar3 is required for coupling dynamic microtubule plus ends to the cortical shmoo tip in budding yeast.
The budding yeast shmoo tip is a model system for analyzing mechanisms coupling force production to microtubule plus-end polymerization/depolymerization. Dynamic plus ends of astral microtubules interact with the shmoo tip in mating yeast cells, positioning nuclei for karyogamy. We have used live-cell imaging of GFP fusions to identify proteins that couple dynamic microtubule plus ends to the shmoo tip. We find that Kar3p, a minus end-directed kinesin motor protein, is required, whereas the other cytoplasmic motors, dynein and the kinesins Kip2p and Kip3p, are not. In the absence of Kar3p, attached microtubule plus ends released from the shmoo tip when they switched to depolymerization. Furthermore, microtubules in cells expressing kar3-1, a mutant that results in rigor binding to microtubules [2], were stabilized specifically at shmoo tips. Imaging of Kar3p-GFP during mating revealed that fluorescence at the shmoo tip increased during periods of microtubule depolymerization. These data are the first to localize the activity of a minus end-directed kinesin at the plus ends of microtubules. We propose a model in which Kar3p couples depolymerizing microtubule plus ends to the cell cortex and the Bim1p-Kar9p protein complex maintains attachment during microtubule polymerization. In support of this model, analysis of Bim1p-GFP at the shmoo tip results in a localization pattern complementary to that of Kar3p-GFP.


Cerebrospinal Fluid Proteomics For Identification Of α2-Macroglobulin As A Potential Biomarker To Monitor Pharmacological Therapeutic Efficacy In Dopamine Dictated Disease States Of Parkinson's Disease And Schizophrenia.
Parkinson's disease and schizophrenia are clinical end points of dopaminergic deficit and excess, respectively, in the mid-brain. In accordance, current pharmacological interventions aim to restore normal dopamine levels, the overshooting of which culminates in adverse effects which results in psychotic symptoms in Parkinson's disease and extra-pyramidal symptoms in schizophrenia. Currently, there are no laboratory assays to assist treatment decisions or help foresee these drug side-effect outcomes. Therefore, the aim was to discover a protein biomarker that had a varying linear expression across the clinical dopaminergic spectrum. iTRAQ-based proteomic experiments along with mass spectrometric analysis was used for comparative proteomics using cerebrospinal fluid (CSF). CSF fluid was collected from 36 patients with Parkinson's disease, 15 patients with urological diseases that served as neurological controls, and seven schizophrenic patients with hallucinations. Validation included ELISA and pathway analysis to highlight the varying expression and provide plausible molecular pathways for differentially expressed proteins in the three clinical phenotypes. Protein profiles were delineated in CSF from Parkinson's disease patients, neurological control and schizophrenia, respectively. Ten of the proteins that were identified had a linear relationship across the dopaminergic spectrum. α-2-Macroglobulin showed to be having high statistical significance on inter-group comparison on validation studies using ELISA. Non-gel-based proteomic experiments are an ideal platform to discover potential biomarkers that can be used to monitor pharmaco-therapeutic efficacy in dopamine-dictated clinical scenarios. α-2 Macroglobulin is a potential biomarker to monitor pharmacological therapy in Parkinson's disease and schizophrenia.


Spectrum of commercial sex activity in Burkina Faso: classification model and risk of exposure to HIV.
Before designing a sexually transmitted infection (STI)/HIV intervention study targeting female commercial sex workers in Bobo Dioulasso, Burkina Faso, we conducted a socioanthropologic survey to analyze the prostitution network in the city in 1998. According to social characteristics, women were classified in six different categories, including four groups of nonprofessional sex workers. The aim of the current study is to assess HIV exposure across this classification model. A total of 447 women belonging to the six categories were enrolled in the study. After collection of social and behavioral data by means of a questionnaire, each woman received a physical examination and a blood sample was taken for HIV serologic testing. The category of "seaters" was the most often infected, with an HIV prevalence of 57% (58 of 101 women). Nonprofessional "sellers" and "bar waitresses" were more often infected than professional "roamers," with an HIV prevalence of 37% (24 of 65 women), 40% (27 of 67 women), and 29% (27 of 92 women), respectively, despite a much lower number of clients per week (average of 2.6 clients, 3.3 clients, and 18.6 clients, respectively). Finally, "students" and "cabarets" (women making and selling local beer in huts) were infected with an HIV prevalence of 15% (9 of 62 women and 9 of 60 women, respectively), which remains higher than the prevalence measured recently in the general female population in the city (6.4%). Our results highlight the high level of vulnerability of nonprofessional sex workers, who need to be considered in the design of any program targeting this population for STI/HIV control purposes.


Mature results of MM-011: a phase I/II trial of liposomal doxorubicin, vincristine, dexamethasone, and lenalidomide combination therapy followed by lenalidomide maintenance for relapsed/refractory multiple myeloma.
A previous interim report of MM-011, the first study that combined lenalidomide with anthracycline-based chemotherapy followed by lenalidomide maintenance for relapsed and/or refractory multiple myeloma (RRMM), showed promising safety and activity. We report the long-term outcomes of all 76 treated patients with follow-up ≥ 5 years. This single-center phase I/II study administered lenalidomide (10 mg on days 1-21 of every 28-day cycle), intravenous liposomal doxorubicin (40 mg/m(2) on day 1), dexamethasone (40 mg on days 1-4), and intravenous vincristine (2 mg on day 1). After 4-6 planned induction cycles, lenalidomide maintenance therapy was given at the last tolerated dose until progression, with or without 50 mg prednisone every other day. The median number of previous therapies was 3 (range, 1-7); 49 (64.5%) patients had refractory disease. Forty-three (56.6%) patients received maintenance therapy. Grade 3/4 adverse events occurred during induction and maintenance therapy in 48.7% and 25.6% of patients, respectively. Four (5.3%) treatment-related deaths occurred during induction. Responses were seen in 53.0% (at least partial response) and 71.2% (at least minor response) of patients. Overall, median progression-free survival and overall survival were 10.5 and 19.0 months, respectively; in patients with refractory disease these values were 7.5 and 11.3 months, respectively. Lenalidomide with anthracycline-based chemotherapy followed by maintenance lenalidomide provided durable control in patients with RRMM (ClinicalTrials.gov number, NCT00091624).


[Search of antimeasles antibodies in HIV-infected children after basic immunization]
OBJECTIVE: To determine the presence of antimeasles antibodies in children perinatally infected with HIV and properly immunized. METHODS: A retrospective cohort study conducted in Belo Horizonte by the Universidade Federal de Minas Gerais, between 1995 and 1996. Twenty one children perinatally infected with HIV and 29 immunocompetent noninfected children were included in the study. Information about measles vaccination was obtained from patientś immunization charts. The presence of neutralizing antibodies against the measles was determined by the plaque reduction neutralization test and IgM was measured by ELISA. The level of significance was set at 5% in all the performed statistical analyses. RESULTS: Median age was 44.5 months for HIV-infected patients and 62.0 months for noninfected children (P=0.64). Both groups received on average two doses of antimeasles vaccine. All HIV-seronegative patients presented antimeasles antibody titers greater than 50 mIU/ml, whereas 57.1% of infected children presented titers above this value (P=0.0001). The geometric mean titer of neutralizing antibodies was significantly lower in the group of HIV-infected children (433.5 mIU/ml) than in noninfected children (1,668.1 mIU/ml), P=0.001. All patients in both groups were negative for antimeasles IgM. CONCLUSION: In the present study, HIV-infected children showed a lower seroprevalence of antimeasles antibody after immunization than noninfected children. These results emphasize the risk of acquisition of measles virus and the need to evaluate alternatives to the vaccination of HIV-infected children in an attempt to maximize the protection against the measles in this group of patients.


Cross-Cultural Contact and Norwegian Language Skills Among Ethnic Minority Women in Norway, and Relationship with Physical Activity in Pregnancy and Postpartum: The STORK-Groruddalen Cohort Study.
There are few studies of the migration context factors and physical activity (PA) level among minority ethnic women in Europe. We investigated the association between migration context factors and moderate to vigorous physical activity (MVPA) among minority ethnic women. Objectively recorded MVPA were obtained from 487 minority ethnic women included in the STORK-Groruddalen Cohort Study at three time points in pregnancy/postpartum. We investigated the associations between (a) contact with ethnic Norwegians and (b) Norwegian language skills and. No associations were observed in pregnancy. Postpartum, women who reported contact with ethnic Norwegians accumulated 17 MVPA min/day (95% CI: -.60, 34.54) more than women with no contact. In complete case analyses, this difference was significant (27 MVPA min/day (95% CI: 8.60, 44.54)). In early postpartum women with contact with ethnic Norwegians seems to be more physically active than women without contact. No associations were observed in pregnancy.


Management of Ewing Sarcoma Family of Tumors: A Short Description of a Rare Primitive Uterine pPNET and Literature Review.
To describe the outcome of a patient with a rare primitive uterine pPNET and to perform a review of the available data in literature, leading the clinicians to better face this rare disease. We have rescued data regarding the multidisciplinary treatment of pPNET from the PUBMED database, highlighting also issues regarding the pathogenesis and the genetic landscape of the ESFTs (Ewing Sarcoma Family of Tumors). Ewing sarcoma and primitive neuroectodermal tumors (PNETs) are small round cell tumors presenting with different degrees of neuroectodermal differentiation. PNETs are further divided into central PNET and peripheral PNET (pPNET). Since pPNETs share the same genetic background of Ewing Sarcomas, they are considered to belong to the Ewing Sarcoma Family of Tumors (ESFTs). Multimodality treatment currently represents the best choice to offer to the affected patients. Although pPNETs are generally diagnosed in children and young adults, an elderly woman aged 85 years came to our attention after a diagnosis of uterine pPNET. Her medical history is presented here, along with a literature review of the subject, highlighting the main biological, pathological and clinical features, with a hypothesis about the possible future therapeutic approaches for these rare malignancies.


CHOPER filters enable rare mutation detection in complex mutagenesis populations by next-generation sequencing.
Next-generation sequencing (NGS) has revolutionized genetics and enabled the accurate identification of many genetic variants across many genomes. However, detection of biologically important low-frequency variants within genetically heterogeneous populations remains challenging, because they are difficult to distinguish from intrinsic NGS sequencing error rates. Approaches to overcome these limitations are essential to detect rare mutations in large cohorts, virus or microbial populations, mitochondria heteroplasmy, and other heterogeneous mixtures such as tumors. Modifications in library preparation can overcome some of these limitations, but are experimentally challenging and restricted to skilled biologists. This paper describes a novel quality filtering and base pruning pipeline, called Complex Heterogeneous Overlapped Paired-End Reads (CHOPER), designed to detect sequence variants in a complex population with high sequence similarity derived from All-Codon-Scanning (ACS) mutagenesis. A novel fast alignment algorithm, designed for the specified application, has O(n) time complexity. CHOPER was applied to a p53 cancer mutant reactivation study derived from ACS mutagenesis. Relative to error filtering based on Phred quality scores, CHOPER improved accuracy by about 13% while discarding only half as many bases. These results are a step toward extending the power of NGS to the analysis of genetically heterogeneous populations.


WASP-related proteins, Abi1 and Ena/VASP are required for Listeria invasion induced by the Met receptor.
Internalisation of the pathogenic bacterium Listeria monocytogenes involves interactions between the invasion protein InlB and the hepatocyte growth factor receptor, Met. Using colocalisation studies, dominant-negative constructs and small interfering RNA (siRNA), we demonstrate a cell-type-dependent requirement for various WASP-related proteins in Listeria entry and InlB-induced membrane ruffling. The WAVE2 isoform is essential for InlB-induced cytoskeletal rearrangements in Vero cells. In HeLa cells, WAVE1, WAVE2 and N-WASP cooperate to promote these processes. Abi1, a key component of WAVE complexes, is recruited at the entry site in both cell types and its inactivation by RNA interference impairs InlB-mediated processes. Ena/VASP proteins also play a role in Listeria internalization, and their deregulation by sequestration or overexpression, modifies actin cups beneath entering particles. Taken together, these results identify the WAVE complex, N-WASP and Ena/VASP as key effectors of the Met signalling pathway and of Listeria entry and highlight the existence of redundant and/or cooperative functions among WASP-family members.


Characterization of clinically significant strains of coagulase-negative staphylococci.
On occasion, a patient may have two or more clinical cultures yielding a coagulase-negative staphylococcus If these multiple isolates have the same phenotype, one might conclude that the same strain was reisolated from the patient, indicating its persistent and pathological presence. We examined the validity of this conclusion when we applied a number of characterizing systems to a collection of 143 isolates of coagulase-negative staphylococci collected during an outbreak of intravascular catheter-associated sepsis. The probability of classifying two random isolates as the same phenotype or species was as follows: P = 0.356 for phage typing, P = 0.348 for Baird-Parker biotyping, P = 0.346 for the API STAPH-IDENT (Analytab Products) system, P = 0.327 for Bentley et al. biotyping, and P = 0.077 for antimicrobial susceptibility patterns. Although antimicrobial susceptibility patterns had the lowest probability, a variability in test results of 7.7% and a tendency for strains to have similar antibiograms effectively raised the probability to P = 0.897. The combination of the API STAPH-IDENT with antibiograms resulted in a probability of P = 0.037 to P = 0.147. When all of the above methods were used together a probability of P = 0.014 was achieved. Five patients had isolates from two or more blood cultures spaced more than 1 day apart that were identical by all of the above criteria, thus confirming prolonged bacteremia. The collection was also examined for the incidence of slime production. Slime production was not associated with any of the above groups, but was associated with symptomatic infections (P less than 0.05) and gentamicin resistance (P less than 0.01). Slime production was strain stable and was of assistance in typing strains of coagulase-negative staphylococci.


Altered phenotype in peripheral blood and tumor-associated NK cells from colorectal cancer patients.
Despite NK cells being originally identified because of their ability to kill tumor cells in vitro, only limited information is available on NK cells infiltration of malignant tumors, especially in humans. NK cells infiltrating human colorectal carcinomas (CRCs) were analyzed to identify their potential protective role in an antitumor immune response. The expression and function of relevant molecules were analyzed from different sources, comparing tumor-associated NK cells (TANKs) with autologous peripheral blood NK cells (PB-NKs) from CRC patients-the latter in comparison with PB-NKs from normal donors. TANKs displayed a profound alteration of their phenotype with a drastic reduction of NK cell receptor expression. Co-culture experiments showed that CRC cells produce modulation in NK phenotype and functionality. Moreover, PB-NKs from CRC patients also exhibited an altered phenotype and profound defects in the ability to activate degranulation and IFN-γ production. For the first time, TANK and PB-NK cells from CRC patients have been characterized. It is shown that they are not capable of producing relevant cytokines and degranulate. Taken together, our results suggest that NK cells from CRC patients present alterations of phenotype and function therefore supporting the progression of cancer.