Lox'd in translation: contradictions in the nomenclature surrounding common lox-site mutants and their implications in experiments.
The Cre-Lox system is a highly versatile and powerful DNA recombinase mechanism, mainly used in genetic engineering to insert or remove desired DNA sequences. It is widely utilized across multiple fields of biology, with applications ranging from plants, to mammals, to microbes. A key feature of this system is its ability to allow recombination between mutant lox sites. Two of the most commonly used mutant sites are named lox66 and lox71, which recombine to create a functionally inactive double mutant lox72 site. However, a large portion of the published literature has incorrectly annotated these mutant lox sites, which in turn can lead to difficulties in replication of methods, design of proper vectors and confusion over the proper nomenclature. Here, we demonstrate common errors in annotations, the impacts they can have on experimental viability, and a standardized naming convention. We also show an example of how this incorrect annotation can induce toxic effects in bacteria that lack optimal DNA repair systems, exemplified by Mycoplasma pneumoniae.


[Adverse effect incidence of Pfizer/BioNTech COVID-19 vaccine: Retrospective cohort study].
Coronavirus SARS-CoV-2 has been the cause of multiple hospitalizations due to respiratory distress, pneumonia, multi-organic failure and death in all the world. The BioNTech/Pfizer lab was the first to get the approval from the Food and Drug Administration (FDA) for vaccine application, beginning with health workers. However, there are a few studies about adverse effects after applying the COVID-19 vaccine. To assess the presence and time of onset of adverse effects produced by the COVID-19 vaccine. A retrospective cohort study was carried out, including medical residents from the Hospital de Especialidades No. 1 (Specialties Hospital No. 1) from the Bajío National Medical Center, who received two doses of Pfizer-BioNTech vaccine and decided to participate voluntarily. We obtained the information by a self-assessment tool sent via email. All adverse effects were collected emphasizing those described in the literature. A total of 101 medical residents were included. It was observed that 55.54% presented adverse effects after the first dose, which appeared in the first 24 hours after the vaccine was administered. After the second dose, 54.5% presented symptomatology, 41.6% during first 24 hours after the vaccine was administered. It was observed symptomatology after applying the complete series (two doses) of BioNTech/Pfizer; however, there were mild and of short duration.


Punctuated plastome reduction and host-parasite horizontal gene transfer in the holoparasitic plant genus Aphyllon.
Foundational studies of chloroplast genome (plastome) evolution in parasitic plants have focused on broad trends across large clades, particularly among the Orobanchaceae, a species-rich and ecologically diverse family of root parasites. However, the extent to which such patterns and processes of plastome evolution, such as stepwise gene loss following the complete loss of photosynthesis (shift to holoparasitism), are detectable at shallow evolutionary time scale is largely unknown. We used genome skimming to assemble eight chloroplast genomes representing complete taxonomic sampling of Aphyllon sect. Aphyllon, a small clade within the Orobanchaceae that evolved approximately 6 Ma, long after the origin of holoparasitism. We show substantial plastome reduction occurred in the stem lineage, but subsequent change in plastome size, gene content, and structure has been relatively minimal, albeit detectable. This lends additional fine-grained support to existing models of stepwise plastome reduction in holoparasitic plants. Additionally, we report phylogenetic evidence based on an rbcL gene tree and assembled 60+ kb fragments of the Aphyllon epigalium mitochondrial genome indicating host-to-parasite horizontal gene transfers (hpHGT) of several genes originating from the plastome of an ancient Galium host into the mitochondrial genome of a recent common ancestor of A. epigalium Ecologically, this evidence of hpHGT suggests that the host-parasite associations between Galium and A. epigalium have been stable at least since its subspecies diverged hundreds of thousands of years ago.


Cell type-specific modulation of fibronectin adhesion functions on chemically-derivatized self-assembled monolayers.
Cell type-specific responses (microfilament stress fibers for fibroblasts or neurites for neuroblastoma cells) were evaluated in culture on inert and chemically-derivatized silane substrata adsorbed with fibronectin (Fn). Substrata of self-assembled monolayers contain a 14-17 carbon aliphatic chain terminating with different chemical endgroups -- [CH3], [C=C], [Br], [CN], [Diol], [COOH], [NH2], [SH], [SCOCH3], or [SO3H]. Fn adsorbed effectively to all derivatized surfaces. 3T3 fibroblasts or neuroblastoma cells attached equivalently to all surfaces preadsorbed with Fn, indicating availability of receptor binding sites on Fns. However, transmembrane signaling from Fn(adsorbed): receptor(cell) surface complexes yielded a range of abilities for generating F-actin stress fibers in fibroblasts or neurites in neuroblastoma cells. Efficiency for stress fiber formation was very different from that of neurite extension. The same chemical endgroups on glass, titanium, or germanium yielded the same patterns of cellular physiological responses, indicating that inert substrata do not act at a distance and that only chemical endgroups regulate Fn signaling functions. When adhesion-inert albumin is co-adsorbed with Fn, efficiency of neurite extension is improved on some surfaces or diminished on others. These results indicate that the conformation of Fn(adsorbed) changes in specific ways on derivatized substrata. Change in Fn conformation was confirmed by FTIR/ATR spectroscopy experiments of Fn(adsorbed). Overall, these studies indicate changes in Fn conformations on chemically-derivatized self-assembled monolayers leading to up- or down-regulation of cell type-specific physiological responses from receptors via their signaling pathways. They also offer predictability for regulating responses of specific cell types when these cells interact with biomaterial implants in vivo.


[Circular shunt in the severe neonatal form of Ebstein's Anomaly. The prostaglandine infusion is it beneficial or harmful?]
Ebstein's disease with functional pulmonary atresia is a severe neonatal presentation of Ebstein's anomaly where the therapeutic management is typically based on the prescription of prostaglandins. The circular shunt is a serious "hemodynamic" complication which is often undiagnosed leading to the discontinuation of prostaglandins. We report a severe neonatal form of Ebstein's anomaly with hemodynamic deterioration relatted to a circular shunt. The diagnosis of Ebstein's anomaly with functional pulmonary atresia was made prenatally at 36 weeks of pregnancy. The patient was born at 38 weeks of gestation by caesarean section. Postnatal ultrasound confirmed the diagnosis. Treatment with prostaglandins was originally created to maintain the vital ductus arteriosus patent. Despite this treatment, hemodynamic deterioration was observed. Ultrasound monitoring showed pictures for a circular shunt. Indeed, blood coming into the pulmonary artery by the wide ductus arteriosus, was "drawn" to the right ventricle and the right atrium due to tricuspid regurgitation and from there to the left heart via the fossa ovalis shunting right to left, when it was ejected into the aorta and the ductus arteriosus. Before this circular shunt, treatment with prostaglandin was discontinued and treatment to reduce pulmonary resistance was described. However, the patient died prior to initiation of treatment. The neonatal form of Ebstein's anomaly is a severe form that can be complicated by a circular shunt. This hemodynamic phenomenon encourages early closure of the ductus arteriosus against indicating the prescription of prostaglandins.


Quantitative multicolor super-resolution microscopy reveals tetherin HIV-1 interaction.
Virus assembly and interaction with host-cell proteins occur at length scales below the diffraction limit of visible light. Novel super-resolution microscopy techniques achieve nanometer resolution of fluorescently labeled molecules. The cellular restriction factor tetherin (also known as CD317, BST-2 or HM1.24) inhibits the release of human immunodeficiency virus 1 (HIV-1) through direct incorporation into viral membranes and is counteracted by the HIV-1 protein Vpu. For super-resolution analysis of HIV-1 and tetherin interactions, we established fluorescence labeling of HIV-1 proteins and tetherin that preserved HIV-1 particle formation and Vpu-dependent restriction, respectively. Multicolor super-resolution microscopy revealed important structural features of individual HIV-1 virions, virus assembly sites and their interaction with tetherin at the plasma membrane. Tetherin localization to micro-domains was dependent on both tetherin membrane anchors. Tetherin clusters containing on average 4 to 7 tetherin dimers were visualized at HIV-1 assembly sites. Combined biochemical and super-resolution analysis revealed that extended tetherin dimers incorporate both N-termini into assembling virus particles and restrict HIV-1 release. Neither tetherin domains nor HIV-1 assembly sites showed enrichment of the raft marker GM1. Together, our super-resolution microscopy analysis of HIV-1 interactions with tetherin provides new insights into the mechanism of tetherin-mediated HIV-1 restriction and paves the way for future studies of virus-host interactions.


Clinical utility of circulating TWEAK and CD163 as biomarkers of iron-induced cardiac decompensation in transfusion dependent thalassemia major.
Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) affects most of the cells involved in cardiac fibrosis like inflammatory cells, cardiomyocytes and fibroblasts. CD163, the receptor of TWEAK on the surface of type 2 macrophages, is shed into plasma upon macrophages activation. This work aimed to evaluate serum TWEAK and its decoy receptor CD163 as probable biomarkers to monitor myocardial iron overload (MIO) in transfusion dependent thalassemia major (TDTM) patients and to predict iron-induced cardiac decompensation (IICD). A total of 140 TDTM patients were enrolled. Patients were categorized into two groups; group I (n = 70) diagnosed with IICD while group II (n = 70) had no evidence of IICD. sTWEAK and sCD163 were quantitated utilizing Enzyme-linked-immunosorbent- assay. sTWEAK was evidently lower in group I than group II (medians, 412 and 1052 pg/mL respectively). sCD163 was higher in group I than group II (medians, 615.5 and 323.5 ng/mL respectively). sTWEAK positively correlated with cardiac MRI-T2 mapping and ventricular ejection fractions and negatively correlated with B-Natriuretic peptide and cardiac troponin. An inverse relationship between TWEAK and CD163 was documented throughout the study. sTWEAK, sCD163 and TWEAK/CD163 ratio proved to be significant predictors of IICD in TDTM patients. TWEAK/CD163 ratio < 1.04 discriminated IICD in TDTM patients with 100 % clinical sensitivity and specificity. Circulating TWEAK and CD163 appears to be promising biomarkers for monitoring MIO and predicting IICD in TDTM patients.


Role of Gut Microbiota, Immune Imbalance, and Allostatic Load in the Occurrence and Development of Diabetic Kidney Disease.
Diabetic kidney disease (DKD) is a prevailing complication arising from diabetes mellitus. Unfortunately, there are no trustworthy and efficacious treatment modalities currently available. In recent times, compelling evidence has emerged regarding the intricate correlation between the kidney and the gut microbiota, which is considered the largest immune organ within the human physique. Various investigations have demonstrated that the perturbation of the gut microbiota and its associated metabolites potentially underlie the etiology and progression of DKD. This phenomenon may transpire through perturbation of both the innate and the adaptive immunity, leading to a burdensome allostatic load on the body and ultimately culminating in the development of DKD. Within this literature review, we aim to delve into the intricate interplay between the gut microbiota, its metabolites, and the immune system in the context of DKD. Furthermore, we strive to explore and elucidate potential chemical interventions that could hold promise for the treatment of DKD, thereby offering invaluable insights and directions for future research endeavors.


Use of a negative selectable marker for rapid selection of recombinant vaccinia virus.
Vaccinia virus has been a powerful tool in molecular biology and vaccine development. The relative ease of inserting and expressing foreign genes combined with its broad host range has made it an attractive antigen delivery system against many heterologous diseases. Many different approaches have been developed to isolate recombinant vaccinia virus generated from homologous recombination; however, most are time-consuming, often requiring a series of passages or specific cell lines. Herein we introduce a rapid method for isolating recombinants using the antibiotic coumermycin and the interferon-associated PKR pathway to select for vaccinia virus recombinants. This method uses a negative selection marker in the form of a fusion protein, GyrB-PKR, consisting of the coumermycin dimerization domain of Escherichia coli gyrase subunit B fused to the catalytic domain of human PKR. Coumermycin-dependent dimerization of this protein results in activation of PKR and the phosphorylation of translation initiation factor, eIF2α. Phosphorylation of this factor leads to an inhibition of protein synthesis, and an inhibition of virus replication. In the presence of coumermycin, recombinants are isolated due to the loss of this coumermycin-sensitive gene by homologous recombination. We demonstrate that this method of selection is highly efficient and requires limited rounds of enrichment to isolate recombinant virus.


Improved effector functions of a therapeutic monoclonal Lewis Y-specific antibody by glycoform engineering.
The aim of the present study was to produce glycosylation variants of the therapeutic Lewis Y-specific humanized IgG1 antibody IGN311 to enhance cell-killing effector function. This was achieved via genetic engineering of the glycosylation machinery of the antibody-producing host. Antibody genes were transiently cotransfected with acetyl-glycosaminyltransferase-III genes into human embryonic kidney-EBV nuclear antigen cells. A control wild-type antibody, IGN311wt, was expressed in the same host using identical expression vectors, but without cotransfection of genes for acetyl-glycosaminyltransferase-III expression. Both expression products were purified to homogeneity and characterized. The glyco-engineered expression product (IGN312-Glyco-I) showed a remarkably homogenous N-linked glycosylation pattern consisting of one major hybrid-type, non-fucosylated and agalactosylated form carrying a bisecting GlcNAc-group. Wild-type expression product (IGN311wt) on the other hand was glycosylated by a multitude of different core-fucosylated complex-type structures of variable degrees of galactosylation. Target affinity of the glyco-engineered antibody as well as heavy and light chain assembly were not affected by acetyl-glycosaminyltransferase-III expression. In vitro experiments showed a approximately 10-fold increase of antibody-dependent cellular cytotoxicity of the glyco-engineered antibody using different Lewis Y-positive target cancer cell lines (SK-BR-3, SK-BR-5, OVCAR-3, and Kato-III). Complement-mediated cytotoxicity of IGN312-Glyco-I was 0.4-fold reduced using SK-BR-5 as target cell line. The reduction of complement activation could be prevented and even converted into a slight increase of activity by using a different molecular-biological approach directing the glycosylation towards increased levels of complex N-linked oligosaccharides of bisected, non-fucosylated type, as a result of cotransfection of mannosidase II together with acetyl-glycosaminyltransferase-III.


Natural and experimental pox virus infection in the domestic cat.
Three naturally occurring cases of cowpox virus infection in the domestic cat are described. Isolate L97 was identified as cowpox virus on the basis of morphology, serology and characteristic cytopathic effect in tissue culture and on the chorioallantoic membrane of embryonated eggs. All three cases showed multiple skin lesions, slight conjunctivitis or purulent ocular discharge but there were no respiratory signs. Two animals recovered, the third was put down as a stray. The disease was reproduced in experimental cats. Isolate L97 was inoculated into two cats intravenously and two cats by skin scarification. All four developed skin lesions at the site or sites of inoculation, and in one cat multiple lesions developed. The two intravenously inoculated animals also developed severe oedema of the neck and brisket around the site of inoculation into the jugular vein, and one cat died. Serological and pathological findings on both the natural and experimental infections are described. Serum neutralising antibody titres in both natural and experimental early convalescent cases were significantly enhanced by the addition of complement.


Epstein-Barr virus and human herpesvirus 8 prevalence in human immunodeficiency virus-associated oral mucosal lesions.
The prevalence of Epstein-Barr virus (EBV) and the recently identified Kaposi's sarcoma (KS)-associated herpesvirus (also designated human herpesvirus 8 [HHV-8]) was determined in oral lesions and oral neoplasms common to persons with human immunodeficiency virus (HIV) infection. Oral lesions were examined by polymerase chain reaction (PCR) for EBV and HHV-8 DNA and by Southern blot analysis for EBV clonality. EBV was detected by Southern blot in hairy leukoplakia lesions, in a subset of AIDS-related lymphomas, and in saliva from HIV-positive persons but not in pseudohairy leukoplakia lesions, oral aphthous ulcers, or oral KS lesions. EBV was detected, however, by PCR in most of the lesions, while HHV-8 was detected only in oral KSs. The absence of HHV-8 DNA in both the EBV-associated hairy leukoplakia lesions and in the EBV-associated AIDS-related lymphomas strengthens the etiologic relationship of EBV to these pathologies and the etiologic role of HHV-8 in KS.


Mispolarization of desmosomal proteins and altered intercellular adhesion in autosomal dominant polycystic kidney disease.
Polycystin-1, the product of the major gene mutated in autosomal dominant polycystic kidney disease (ADPKD), has been shown to associate with multiple epithelial cell junctions. Our hypothesis is that polycystin-1 is an important protein for the initial establishment of cell-cell junctions and maturation of the cell and that polycystin-1 localization is dependent on the degree of cell polarization. Using laser-scanning confocal microscopy and two models of cell polarization, polycystin-1 and desmosomes were found to colocalize during the initial establishment of cell-cell contact when junctions were forming. However, colocalization was lost in confluent monolayers. Parallel morphological and biochemical evaluations revealed a profound mispolarization of desmosomal components to both the apical and basolateral domains in primary ADPKD cells and tissue. Studies of the intermediate filament network associated with desmosomes showed that there is a decrease in cytokeratin levels and an abnormal expression of the mesenchymal protein vimentin in the disease. Moreover, we show for the first time that the structural alterations seen in adherens and desmosomal junctions have a functional impact, leaving the ADPKD cells with weakened cell-cell adhesion. In conclusion, in this paper we show that polycystin-1 transiently colocalizes with desmosomes and that desmosomal proteins are mislocalized as a consequence of polycystin-1 mutation.


Murder-suicide of the jealous paranoia type: a multicenter statistical pilot study.
The authors present a pilot statistical study of murder-suicide comprising 32 cases from the years 1990-1992, collected from the offices of the medical examiners of seven counties in five of the United States. The study includes brief reviews of previous statistical surveys of murder, murder-suicide, and suicide. This present study's conclusions parallel the findings of previous research on the demographic characteristics of the perpetrators of murder-suicide, the relationship between killers and victims, the types of weapon used, locations of the incidents, and the time intervals between the murder and suicide. It also highlights the similarities between the characteristics of the perpetrator of murder-suicide and those of persons who commit only suicide, supporting the thesis that murder-suicide is an extended suicide. Suggestions for prevention of such a type of crime are offered.


Survey on the current gynaecological approach of ovarian cancer patients: The utility of HIPEC.
The aim of this survey was to acquire an overview of the current management of ovarian cancer with an emphasis on the utility of hyperthermic intraperitoneal chemotherapy (HIPEC).Methods: An email was sent to Oncologists prior to PSOGI International Symposium on Advanced Ovarian Cancer, Athens 11-13 April 2019. Doctors submitted responses on the relevant website. The self-report survey contained 17 questions. In total, 467 Medical Oncologists, Surgical Oncologists or Gynaecologic Oncologists were participated and answered to this survey. The resectability of disease was evaluated by laparoscopy from 48.5% of the participants, while 51.5% answered that they stage their patients pre-surgically with the use of CT or MRI. The preferred first intervention in advanced ovarian cancer patients is the neoadjuvant chemotherapy followed by interval cytoreductive surgery (72%). Regarding the use of HIPEC, almost half of the participants answered that there is role of HIPEC use in ovarian cancer patients undergoing interval debulking surgery, while almost 70% answered positively about the utility of HIPEC use in ovarian cancer recurrence. As for the role of lymphadenectomy in advanced ovarian cancer patients, half of the responders answered negatively. Finally, only 25% of the participants responded that they always check the BRCA status of their ovarian cancer patients, despite the possible differentiation of treatment based on the molecular profiling (80%). The results of this survey indicate the utility of HIPEC in treatment of ovarian cancer patients and the differences in the overall management of ovarian cancer patients in the current clinical practice.


Is aging raw cattle urine efficient for sampling Anopheles arabiensis Patton?
To ensure sustainable routine surveillance of mosquito vectors, simple, effective and ethically acceptable tools are required. As a part of that, we evaluated the efficiency of resting boxes baited with fresh and aging cattle urine for indoor and outdoor sampling of An. arabiensis in the lower Moshi rice irrigation schemes. A cattle urine treatment and re-treatment schedule was used, including a box with a piece of cloth re-treated with urine daily, and once after 3 and 7 day. Resting box with piece of black cloth not treated with urine was used as a control. Each treatment was made in pair for indoor and outdoor sampling. A 4 by 4 Latin square design was used to achieve equal rotation of each of the four treatments across the experimental houses. Sampling was done over a period of 6 months, once per week. A total of 7871 mosquitoes were collected throughout the study period. 49.8% of the mosquitoes were collected from resting box treated with urine daily; 21.6% and 20.0% were from boxes treated 3 and 7 days respectively. Only 8.6% were from untreated resting box (control). The proportion collected indoors was approximately 2 folds greater than the outdoor. Of all mosquitoes, 12.3% were unfed, 4.1% full fed, 34.2% semi-gravid and 49.4% gravid. Fresh and decaying cattle urine odour baited resting boxes offer an alternative tool for sampling particularly semi-gravid and gravid An. arabiensis. Evaluation in low density seasons of An. arabiensis in different ecological settings remains necessary. This sampling method may be standardized for replacing human landing catch.


Sport nutrition for young athletes.
Nutrition is an important part of sport performance for young athletes, in addition to allowing for optimal growth and development. Macronutrients, micronutrients and fluids in the proper amounts are essential to provide energy for growth and activity. To optimize performance, young athletes need to learn what, when and how to eat and drink before, during and after activity.


Forced use as a home-based intervention in children with congenital hemiplegic cerebral palsy: choosing the appropriate constraint.
The purpose of this study was (1) to identify the most appropriate splint from children's and parents' perspective as reflected by effectiveness and adherence to home-based forced use therapy (FUT). (2) To provide guidance in the development of a practical and effective protocol based on forced use principles. A crossover design with a convenience sample of children with hemiplegic cerebral palsy and their parents was used to test three types of splint (mitt, short splint and long splint) during home-based FUT. Children wore the splints for a minimum of 1 h/ day during physical activity. Outcome measures included a daily log and a questionnaire (completed by parents), an actometer worn on the affected hand to quantify movement and video recordings to inform quality of movement. The short splint was found to be the most effective and acceptable restraining device. Wearing devices for more than 1 h per day was not considered acceptable by either parents or children. A short splint, worn for 1 h per day was found to be the most acceptable protocol.


Nerve Growth Factor and Pain Mechanisms.
Nerve growth factor (NGF) antagonism is on the verge of becoming a powerful analgesic treatment for numerous conditions, including osteoarthritis and lower back pain. This review summarizes the historical research, both fundamental and clinical, that led to our current understanding of NGF biology. We also discuss the surprising number of questions that remain about NGF expression patterns and NGF's various functions and interaction partners in relation to persistent pain and the potential side effects of anti-NGF therapy.


Xanthine oxidase-mediated intracellular oxidative stress in response to cerulein in rat pancreatic acinar cells.
Intralobular oxygen radical formation was examined in cerulein-stimulated rat pancreatic acinar cells by digital imaging microscopic fluorography using a hydroperoxide-sensitive fluorescent probe, dichlorofluorescin (DCFH) diacetate. The isolated pancreatic acinar cells loaded with DCFH diacetate were microscopically observed, and the dichlorofluorescein (DCF) fluorescence yielded by DCFH oxidation via hydroperoxides was digitally processed. Within the initial 20 min after the application of cerulein (10 microM), intracellular oxidative stress was observed as indicated by the increase in DCF fluorescence intensity and reached its maximum at 60 min. DCF fluorescence intensity was then gradually decreased until 80 min, followed by a marked increase in propidium iodide (PI) fluorescence, suggesting irreversible cell death. Allopurinol (1 microM), a xanthine oxidase inhibitor, significantly attenuated the early increase of DCF fluorescence intensity as well as the late cell damage. Treatment with hyperbaric oxygen (PO2 300 mm Hg) also significantly attenuated both the increase of DCF fluorescence and the number of PI-positive cells. The results suggest that xanthine oxidase-mediated oxygen radicals may play an important role in cerulein-induced intracellular oxidative stress in pancreatic acinar cells of rats.


Comparison of Marginal Fit and Internal Adaptation of Copings Fabricated with Polyetheretherketone and Zirconia - An In vitro Study.
Purpose: The aim of this in vitro study was to evaluate and compare the Marginal fit and Internal adaptation of copings fabricated with polyetheretherketone (PEEK) and Zirconia luted with resin cements. Maxillary first premolar was prepared to receive all ceramic restoration. It was duplicated using addition silicon impression material for fabricating Cobalt-Chromium metal die, which is used as a master die. Using the master die, 30 heat cure acrylic samples were fabricated for the heat cure acrylic tooth models on which 15 PEEK and 15 zirconia copings were made. Self-adhesive resin cement is used for luting and 30 copings were luted. After 24 h, luted copings were having a disc of thickness 0.01 mm cutting at high speed using a Tooth cutting lathe for all the 30 samples. Field Scanning Electron Microscopic study analysis for evaluating marginal fit and internal adaptation of two groups were observed. The values were then analyzed using one-way ANOVA (post hoc) followed by Dunnet t-test. Among those two groups, PEEK copings group materials showed the lowest mean value of (30.3 ± 5.1) for marginal gap, (29.1 ± 5.8) for internal gap whereas the zirconia copings group showed a mean value of (50.26 ± 16.02) and (32.8 ± 5.2) respectively. Among these two groups, comparatively less marginal fit and internal adaptation is seen in zirconia copings when compared to the PEEK copings. While the marginal fit and internal adaptation of both PEEK and zirconia copings were in the acceptable clinical range.


An inflammatory-related genes signature based model for prognosis prediction in breast cancer.
Breast cancer has become the most common malignant tumor in the world. It is vital to discover novel prognostic biomarkers despite the fact that the majority of breast cancer patients have a good prognosis because of the high heterogeneity of breast cancer, which causes the disparity in prognosis. Recently, inflammatory-related genes have been proven to play an important role in the development and progression of breast cancer, so we set out to investigate the predictive usefulness of inflammatory-related genes in breast malignancies. We assessed the connection between Inflammatory-Related Genes (IRGs) and breast cancer by studying the TCGA database. Following differential and univariate Cox regression analysis, prognosis-related differentially expressed inflammatory genes were estimated. The prognostic model was constructed through the Least Absolute Shrinkage and Selector Operation (LASSO) regression based on the IRGs. The accuracy of the prognostic model was then evaluated using the Kaplan-Meier and Receiver Operating Characteristic (ROC) curves. The nomogram model was established to predict the survival rate of breast cancer patients clinically. Based on the prognostic expression, we also looked at immune cell infiltration and the function of immune-related pathways. The CellMiner database was used to research drug sensitivity. In this study, 7 IRGs were selected to construct a prognostic risk model. Further research revealed a negative relationship between the risk score and the prognosis of breast cancer patients. The ROC curve proved the accuracy of the prognostic model, and the nomogram accurately predicted survival rate. The scores of tumor-infiltrating immune cells and immune-related pathways were utilized to calculate the differences between the low- and high-risk groups, and then explored the relationship between drug susceptibility and the genes that were included in the model. These findings contributed to a better understanding of the function of inflammatory-related genes in breast cancer, and the prognostic risk model provides a potentially promising prognostic strategy for breast cancer.


AnnotCompute: annotation-based exploration and meta-analysis of genomics experiments.
The ever-increasing scale of biological data sets, particularly those arising in the context of high-throughput technologies, requires the development of rich data exploration tools. In this article, we present AnnotCompute, an information discovery platform for repositories of functional genomics experiments such as ArrayExpress. Our system leverages semantic annotations of functional genomics experiments with controlled vocabulary and ontology terms, such as those from the MGED Ontology, to compute conceptual dissimilarities between pairs of experiments. These dissimilarities are then used to support two types of exploratory analysis-clustering and query-by-example. We show that our proposed dissimilarity measures correspond to a user's intuition about conceptual dissimilarity, and can be used to support effective query-by-example. We also evaluate the quality of clustering based on these measures. While AnnotCompute can support a richer data exploration experience, its effectiveness is limited in some cases, due to the quality of available annotations. Nonetheless, tools such as AnnotCompute may provide an incentive for richer annotations of experiments. Database URL: http://www.cbil.upenn.edu/annotCompute/


Airway pressure release ventilation for lung protection in acute respiratory distress syndrome: an alternative way to recruit the lungs.
Airway pressure release ventilation (APRV) is a modality of ventilation in which high inspiratory continuous positive airway pressure (CPAP) alternates with brief releases. In this review, we will discuss the rationale for APRV as a lung protective strategy and then provide a practical introduction to initiating APRV using the time-controlled adaptive ventilation (TCAV) method. APRV using the TCAV method uses an extended inspiratory time and brief expiratory release to first stabilize and then gradually recruit collapsed lung (over hours/days), by progressively 'ratcheting' open a small volume of collapsed tissue with each breath. The brief expiratory release acts as a 'brake' preventing newly recruited units from re-collapsing, reversing the main drivers of ventilator-induced lung injury (VILI). The precise timing of each release is based on analysis of expiratory flow and is set to achieve termination of expiratory flow at 75% of the peak expiratory flow. Optimization of the release time reflects the changes in elastance and, therefore, is personalized (i.e. conforms to individual patient pathophysiology), and adaptive (i.e. responds to changes in elastance over time). APRV using the TCAV method is a paradigm shift in protective lung ventilation, which primarily aims to stabilize the lung and gradually reopen collapsed tissue to achieve lung homogeneity eliminating the main mechanistic drivers of VILI.


Humanin rescues cultured rat cortical neurons from NMDA-induced toxicity through the alleviation of mitochondrial dysfunction.
N-methyl-D-aspartate (NDMA) receptor-mediated excitotoxicity has been implicated in a variety of pathological situations such as Alzheimer's disease (AD) and Parkinson's disease. However, no effective treatments for the same have been developed so far. Humanin (HN) is a 24-amino acid peptide originally cloned from the brain of patients with AD and it prevents stress-induced cell death in many cells/tissues. In our previous study, HN was found to effectively rescue rat cortical neurons. It is still not clear whether HN protects the neurons through the attenuation of mitochondrial dysfunction. In this study, excitatory toxicity was induced by NMDA, which binds the NMDA receptor in primarily cultured rat cortical neurons. We found that NMDA (100 μmol/L) dramatically induced the decrease of cell viability and caused mitochondrial dysfunction. Pretreatment of the neurons with HN (1 μmol/L) led to significant increases of mitochondrial succinate dehydrogenase (SDH) activity and membrane potential. In addition, HN pretreatment significantly reduced the excessive production of both reactive oxygen species (ROS) and nitric oxide (NO). Thus, HN could attenuate the excitotoxicity caused by the overactivation of the NMDA receptor through the alleviation of mitochondrial dysfunction.


Hybrid Dysfunction Expressed as Elevated Metabolic Rate in Male Ficedula Flycatchers.
Studies of ecological speciation are often biased towards extrinsic sources of selection against hybrids, resulting from intermediate hybrid morphology, but the knowledge of how genetic incompatibilities accumulate over time under natural conditions is limited. Here we focus on a physiological trait, metabolic rate, which is central to life history strategies and thermoregulation but is also likely to be sensitive to mismatched mitonuclear interactions. We measured the resting metabolic rate of male collared, and pied flycatchers as well as of naturally occurring F1 hybrid males, in a recent hybrid zone. We found that hybrid males had a higher rather than intermediate metabolic rate, which is indicative of hybrid physiological dysfunction. Fitness costs associated with elevated metabolic rate are typically environmentally dependent and exaggerated under harsh conditions. By focusing on male hybrid dysfunction in an eco-physiological trait, our results contribute to the general understanding of how combined extrinsic and intrinsic sources of hybrid dysfunction build up under natural conditions.


Bacterial tactic response to silver nanoparticles.
In this study, we investigated the tactic response of Pseudomonas putida G7, a representative soil bacterium, towards silver nanoparticles (AgNPs). The study integrated the characterization of surface area and size distribution of AgNPs, toxicity determinations, based on ATP production, and assessment of the repellent reaction by means of an inverted capillary assay ('chemical-in-pond' method), and changes in the motility behaviour determined by computer-assisted motion analysis. Our data demonstrate, for the first time, that nanoparticles can elicit a negative tactic response in bacteria at low but environmentally relevant, sublethal concentrations. Data obtained by the chemical-in-pond method indicated that cells exposed to 0.1 mg l(-1) of two AgNPs preparations, differing in particle size (maximum diameter ≤ 100 nm and ≤ 150 nm respectively), were repelled in the gradients created inside the capillaries. However, cells exposed to similar low concentration of AgNO3 did not demonstrate any detectable repellent response, although it reduced cell viability by 20%, a decrease comparable to that caused by AgNPs. Computer analysis of swimming behaviour of cells exposed to AgNPs (0.2 mg l(-1) ) revealed a significant increase in turning events, as compared with unexposed controls, which is characteristic of bacterial repellent response. Greater AgNPs concentrations (up to 100 mg l(-1) ) also induced changes in the swimming behaviour, although they did not induce any detectable repellent response as determined by the chemical-in-pond assays. In contrast, AgNO3 failed to induce the repellent swimming behaviour within the wide range of concentrations tested (0.001-100 mg l(-1) ), and caused a significant inhibition of cell motility at a concentration above 0.1 mg l(-1) . The evidence presented here suggests there are likely to be alternative mechanisms by which nano-scale silver induces a repellent response, which is more direct than the toxic response of macro-forms of silver, attributed to ion formation and exposure.


Multi-channel silk sponge mimicking bone marrow vascular niche for platelet production.
In the bone marrow, the interaction of progenitor cells with the vasculature is fundamental for the release of blood cells into circulation. Silk fibroin, derived from Bombyx mori silkworm cocoons, is a promising protein biomaterial for bone marrow tissue engineering, because of its tunable architecture and mechanical properties, the capacity to incorporate labile compounds without loss of bioactivity and the demonstrated ability to support blood cell formation without premature activation. In this study, we fabricated a custom perfusion chamber to contain a multi-channel lyophilized silk sponge mimicking the vascular network in the bone marrow niche. The perfusion system consisted in an inlet and an outlet and 2 splitters that allowed funneling flow in each single channel of the silk sponge. Computational Fluid Dynamic analysis demonstrated that this design permitted confined flow inside the vascular channels. The silk channeled sponge supported efficient platelet release from megakaryocytes (Mks). After seeding, the Mks localized along SDF-1α functionalized vascular channels in the sponge. Perfusion of the channels allowed the recovery of functional platelets as demonstrated by increased PAC-1 binding upon thrombin stimulation. Further, increasing the number of channels in the silk sponge resulted in a proportional increase in the numbers of platelets recovered, suggesting applicability to scale-up for platelet production. In conclusion, we have developed a scalable system consisting of a multi-channeled silk sponge incorporated in a perfusion chamber that can provide useful technology for functional platelet production ex vivo.


Overcoming Rooming-In Barriers: A Survey on Mothers' Perspectives.
Background: The importance of rooming-in in promoting breastfeeding initiation and continuation within the 10 Steps for Successful Breastfeeding is widely acknowledged. However, adherence to this practice by healthcare facilities is lower than that of other Steps. A deeper knowledge of maternal rooming-in experience has been advocated to identify the most effective rooming-in policies, thus enabling mothers to have a positive experience when practicing it in the postpartum period. Aim: To investigate maternal knowledge of rooming-in and the most frequently encountered barriers and possible facilitators of adherence to the practice, according to their experience. Study Design and Methods: We enrolled mothers who delivered healthy term or late preterm infants during the month of January 2019 in a tertiary referral center for neonatal care in Milan, Italy. At discharge, a structured interview about mothers' rooming-in experience was administered by healthcare professionals. Basic subjects' characteristics and mode of feeding were recorded. Results: The enrolled population included 328 mothers and 333 neonates. The great majority of mothers knew of rooming-in and 48.2% practiced it continuously. The 86.3% of mothers was aware of the beneficial effects of rooming-in; promotion of mother-infant bonding, increased confidence in taking care of the baby and ability to recognize baby's feeding cues were the most frequently cited, whereas improving breastfeeding was reported by a limited number of mothers, unless they were asked a specific question about it. The main reported obstacles were fatigue (40.5%) and cesarean section related difficulties (15.5%); night was the most critical time of the day for rooming-in. Strategies suggested by mothers for improving rooming-in were increased assistance to the dyad, organizational and structural changes and the possibility to have a family member during the night. Additionally, mothers who adhered to rooming-in practice continuously during hospital stay had a higher exclusive breastfeeding rate at discharge compared to mothers who did not. Conclusions: Our study contributes to a deeper knowledge of maternal rooming-in experience in an Italian tertiary maternity. We underline the importance of providing a tailored support to the mother-infant dyad in order to overcome rooming-in barriers perceived by mothers and promote a positive rooming-in experience.


A novel CASR mutation associated with neonatal severe hyperparathyroidism transmitted as an autosomal recessive disorder.
Neonatal severe primary hyperparathyroidism (NSHPT, MIM 239200) is most often an isolated disorder that is due to biallelic inactivating mutations in the CASR, the gene encoding the calcium sensing receptor; NSHPT is inherited from parents with familial hypocalciuric hypercalcemia, each of whom has one mutated CASR allele. To report clinical and genetic findings in a brother and sister with NSHPT due to a novel mutation in the CASR transmitted as an autosomal recessive trait and to examine the functional effect of the mutation. A brother and sister with marked hypercalcemia due to NSHPT were identified; the boy also had craniosynostosis requiring surgical repair. The genotyping of the CASR in both children and their parents who were eucalcemic and normophosphatemic was undertaken. In order to examine the significance of the variant CASR identified, the CASR variant was expressed in vitro and examined by three computer computational programs [PolyPhen2, MutationTaster, Sorting Intolerant From Tolerant (SIFT)] designed to evaluate the effect of a nucleotide variant on the structure and likely functional consequence upon the protein product. A sequence variant in the CASR was identified [G>T point mutation at nucleotide c.2303 in exon 7 (c.2303G>T) resulting in the replacement of glycine by valine at codon 768 (p.Gly768Val)]. Two copies of this CASR variant were present in the genome of the siblings while a single copy of the CASR variant was present in both of the clinically and biochemically normal parents, a pattern of transmission consistent with autosomal recessive inheritance of NSHPT in this family. When expressed in HEK293 cells in vitro, the novel Gly768Val variant did not interfere with protein generation or migration to the cell membrane in vitro. The analysis of the functional effect of the Gly768Val CASR variant by the PolyPhen2, MutationTaster, and Sorting Intolerant From Tolerant computer programs revealed that this mutation was very likely to be deleterious. The NSHPT associated with biallelic Gly768Val mutations of the CASR in two siblings with severe hypercalcemia and hyperparathyroidism and their clinically and biochemically normal heterozygous parents was transmitted as an autosomal recessive disorder in this family.


Modeling probability and additive summation for detection across multiple mechanisms under the assumptions of signal detection theory.
Many studies have investigated how multiple stimuli combine to reach threshold. There are broadly speaking two ways this can occur: additive summation (AS) where inputs from the different stimuli add together in a single mechanism, or probability summation (PS) where different stimuli are detected independently by separate mechanisms. PS is traditionally modeled under high threshold theory (HTT); however, tests have shown that HTT is incorrect and that signal detection theory (SDT) is the better framework for modeling summation. Modeling the equivalent of PS under SDT is, however, relatively complicated, leading many investigators to use Monte Carlo simulations for the predictions. We derive formulas that employ numerical integration to predict the proportion correct for detecting multiple stimuli assuming PS under SDT, for the situations in which stimuli are either equal or unequal in strength. Both formulas are general purpose, calculating performance for forced-choice tasks with M alternatives, n stimuli, in Q monitored mechanisms, each subject to a non-linear transducer with exponent τ. We show how the probability (and additive) summation formulas can be used to simulate psychometric functions, which when fitted with Weibull functions make signature predictions for how thresholds and psychometric function slopes vary as a function of τ, n, and Q. We also show how one can fit the formulas directly to real psychometric functions using data from a binocular summation experiment, and show how one can obtain estimates of τ and test whether binocular summation conforms more to PS or AS. The methods described here can be readily applied using software functions newly added to the Palamedes toolbox.


Comparison of antinociceptive efficacy and gastroprotection between celecoxib and diclofenac plus misoprostol in rats.
The present study was designed to assess the antinociceptive efficacy and gastroprotective activity of the mixture of diclofenac and misoprostol and its comparison with celecoxib in rats. The effect of diclofenac/misoprostol and celecoxib was assessed in the 1% formalin test. Female Wistar rats were fasted 12 hr before experiments and diclofenac (10 and 50 mg/kg), misoprostol (100 microg/kg), celecoxib (30 and 100 mg/kg), saline and the combination of diclofenac (50 mg/kg) plus misoprostol (25, 50 and 100 microg/kg) were administered orally. Nociceptive behavior was assessed during the following hr. Diclofenac and celecoxib dose-dependently reduced formalin-induced nociception reaching similar maximal effects. Moreover, misoprostol did not produce antinociception, but increased diclofenac-induced antinociception. Animals were sacrificed 3 hr following drug administration and stomachs examined to assess gastric damage. Misoprostol did not produce any damage to the stomach. However, diclofenac, but not celecoxib, produced significant gastric damage (number of stomach ulcers) in a dose-dependent fashion. Misoprostol dose-dependently reduced diclofenac-induced ulcers. Data show that diclofenac and celecoxib lead to similar antinociception, with diclofenac being more active to produce gastric damage. However, diclofenac-induced gastric damage can be markedly reduced by misoprostol. In addition to its gastroprotective effect, misoprostol showed a synergic effect on diclofenac-induced anti-nociception. Considering the cardiovascular effects of COX-2 selective inhibitors, the combination of diclofenac and misoprostol could represent a safer and effective alternative for patients with pain.


Expanding oxygen-minimum zones in the tropical oceans.
Oxygen-poor waters occupy large volumes of the intermediate-depth eastern tropical oceans. Oxygen-poor conditions have far-reaching impacts on ecosystems because important mobile macroorganisms avoid or cannot survive in hypoxic zones. Climate models predict declines in oceanic dissolved oxygen produced by global warming. We constructed 50-year time series of dissolved-oxygen concentration for select tropical oceanic regions by augmenting a historical database with recent measurements. These time series reveal vertical expansion of the intermediate-depth low-oxygen zones in the eastern tropical Atlantic and the equatorial Pacific during the past 50 years. The oxygen decrease in the 300- to 700-m layer is 0.09 to 0.34 micromoles per kilogram per year. Reduced oxygen levels may have dramatic consequences for ecosystems and coastal economies.


Evaluation of Patient Tolerance and Small-Bowel Distention With a New Small-Bowel Distending Agent for Enterography.
The objective of our study was to compare a flavored beverage containing a thickening agent for enterography with a low-Hounsfield-value barium suspension for side effects, taste, subjects' willingness to repeat the drinking protocol, and small-bowel distention. The following five drinking protocols were administered to 10 volunteers: 1000 mL of flavored beverage followed by 350 mL of water, 1500 mL of flavored beverage, 900 mL of low-Hounsfield-value barium suspension followed by 450 mL of water, 1350 mL of low-Hounsfield-value barium suspension followed by 150 mL of water, and 1500 mL of water. MR images were obtained 50 and 60 minutes after initiation of drinking. Subjects completed a questionnaire evaluating the side effects, the taste of the drink, and their willingness to repeat the drinking protocol. Reviewers assigned scores evaluating small-bowel distention and ranked the examinations in order of preference. There was no significant difference in nausea or vomiting among the protocols (p = 0.20 and 0.42, respectively), but larger volumes of flavored beverage and low-Hounsfield-value barium suspension resulted in more cramping and diarrhea (p = 0.001 and 0.002, respectively). The taste of the low-Hounsfield-value barium suspension was rated the worst (p < 0.0001). The subjects' willingness to repeat the drinking protocol was highest for the 1000 mL of flavored beverage or water alone (p < 0.05). There were no significant differences in subjective small-bowel distention except that water was rated the worst by two of the three readers (p < 0.02). There was no significant difference in the diameter of the most dis-tended small bowel for any segment or reader (p > 0.23). A flavored beverage containing a thickening agent has a similar side effect profile and results in equivalent small-bowel distention compared with a low-Hounsfield-value barium suspension, but subjects rate taste and their willingness to repeat the drinking protocol higher for this new agent.


Novel scabies mite serpins inhibit the three pathways of the human complement system.
Scabies is a parasitic infestation of the skin by the mite Sarcoptes scabiei that causes significant morbidity worldwide, in particular within socially disadvantaged populations. In order to identify mechanisms that enable the scabies mite to evade human immune defenses, we have studied molecules associated with proteolytic systems in the mite, including two novel scabies mite serine protease inhibitors (SMSs) of the serpin superfamily. Immunohistochemical studies revealed that within mite-infected human skin SMSB4 (54 kDa) and SMSB3 (47 kDa) were both localized in the mite gut and feces. Recombinant purified SMSB3 and SMSB4 did not inhibit mite serine and cysteine proteases, but did inhibit mammalian serine proteases, such as chymotrypsin, albeit inefficiently. Detailed functional analysis revealed that both serpins interfered with all three pathways of the human complement system at different stages of their activation. SMSB4 inhibited mostly the initial and progressing steps of the cascades, while SMSB3 showed the strongest effects at the C9 level in the terminal pathway. Additive effects of both serpins were shown at the C9 level in the lectin pathway. Both SMSs were able to interfere with complement factors without protease function. A range of binding assays showed direct binding between SMSB4 and seven complement proteins (C1, properdin, MBL, C4, C3, C6 and C8), while significant binding of SMSB3 occurred exclusively to complement factors without protease function (C4, C3, C8). Direct binding was observed between SMSB4 and the complement proteases C1s and C1r. However no complex formation was observed between either mite serpin and the complement serine proteases C1r, C1s, MASP-1, MASP-2 and MASP-3. No catalytic inhibition by either serpin was observed for any of these enzymes. In summary, the SMSs were acting at several levels mediating overall inhibition of the complement system and thus we propose that they may protect scabies mites from complement-mediated gut damage.


Intentional Drug Overdose Involving Pregabalin and Gabapentin: Findings from the National Self-Harm Registry Ireland, 2007-2015.
Intentional drug overdose (IDO) is a significant public health problem. Concerns about the misuse of gabapentinoids, i.e. pregabalin and gabapentin, including their consumption in IDO have grown in recent years. This paper examines the trends in the prevalence of gabapentinoids taken in IDO, the profile of individuals taking them, and associated overdose characteristics. Presentations to emergency departments involving IDO, recorded by the National Self-Harm Registry Ireland between 1 January 2007 and 31 December 2015 were examined. Data items included patient demographics, drug names, total tablet quantity consumed and alcohol involvement. Gabapentinoids were involved in 2115 (2.9%) of the 72,391 IDOs recorded. Presentations involving a gabapentinoid increased proportionally from 0.5% in 2007 to 5.5% in 2015. The majority of IDOs involving a gabapentinoid were made by females (59.9%), with over one-third (37.2%) involving alcohol. Compared with IDOs involving other drugs, presentations with a gabapentinoid were made by persons who were older (median 37 vs. 32 years) and involved a significantly greater median quantity of tablets (30 vs. 21, p ≤ 0.001), with over one-quarter (27.4%) of these involving the ingestion of 50 tablets or more. Admission to hospital was significantly more common following IDOs with a gabapentinoid compared with those without (49.4% vs. 41.4%, p ≤ 0.001). This study identified the increasing use of gabapentinoids in IDO, describing the profile and overdose characteristics of presentations. It is important for clinicians to exercise vigilance while prescribing gabapentinoids, including being aware of other medications that their patients may have access to. Our findings support the need for routine monitoring for signs of misuse among those prescribed gabapentinoids.


Videotapes in evaluating work-related upper extremity symptoms.
Thirteen patients with upper extremity symptoms that were claimed to have occurred in the course of employment were evaluated to determine the role of videotapes in their evaluation and management. Videotapes were of two types: work demonstration by patient or coworker (8 tapes) and surveillance tapes obtained by a private investigator (5 tapes). Four of eight work station videotapes demonstrated significant repetitive motion that could have contributed to their symptoms. Four of eight work station videotapes demonstrated that the tasks were neither forceful nor repetitive in nature. Return to work recommendations were made based on both clinical grounds and job site information provided on tape. After viewing five surveillance videotapes, two fraudulent claims were settled soon after medical opinions were rendered. Two patients were declared able to return to work; one returned to work and the other was dismissed. The videotape of patient No. 13 was not crucial for the decision and he was authorized to have surgery. The opinions formed concerning the causality of alleged claims of injury were often altered by viewing the content of the videotapes. Videotapes are a valuable tool and useful adjunct in the overall management of the workers with upper extremity symptoms.


The deployment of carbon monoxide wireless sensor network (CO-WSN) for ambient air monitoring.
Wireless sensor networks are becoming increasingly important as an alternative solution for environment monitoring because they can reduce cost and complexity. Also, they can improve reliability and data availability in places where traditional monitoring methods are difficult to site. In this study, a carbon monoxide wireless sensor network (CO-WSN) was developed to measure carbon monoxide concentrations at a major traffic intersection near the University of Cincinnati main campus. The system has been deployed over two weeks during Fall 2010, and Summer 2011-2012, traffic data was also recorded by using a manual traffic counter and a video camcorder to characterize vehicles at the intersection 24 h, particularly, during the morning and evening peak hour periods. According to the field test results, the 1 hr-average CO concentrations were found to range from 0.1-1.0 ppm which is lower than the National Ambient Air Quality Standards (NAAQS) 35 ppm on a one-hour averaging period. During rush hour periods, the traffic volume at the intersection varied from 2,067 to 3,076 vehicles per hour with 97% being passenger vehicles. Furthermore, the traffic volume based on a 1-h average showed good correlation (R2 = 0.87) with the 1-h average CO-WSN concentrations for morning and evening peak time periods whereas CO-WSN results provided a moderate correlation (R2 = 0.42) with 24 hours traffic volume due to fluctuated changes of meteorological conditions. It is concluded that the performance and the reliability of wireless ambient air monitoring networks can be used as an alternative method for real time air monitoring.


Robot-assisted therapy for arm recovery for stroke patients: state of the art and clinical implication.
Introduction: Robot-assisted therapy is an emerging approach that performs highly repetitive, intensive, task oriented and quantifiable neuro-rehabilitation. In the last decades, it has been increasingly used in a wide range of neurological central nervous system conditions implying an upper limb paresis. Results from the studies are controversial, for the many types of robots and their features often not accompanied by specific clinical indications about the target functions, fundamental for the individualized neurorehabilitation program.Areas covered: This article reviews the state of the art and perspectives of robotics in post-stroke rehabilitation for upper limb recovery. Classifications and features of robots have been reported in accordance with technological and clinical contents, together with the definition of determinants specific for each patient, that could modify the efficacy of robotic treatments. The possibility of combining robotic intervention with other therapies has also been discussed.Expert commentary: The recent wide diffusion of robots in neurorehabilitation has generated a confusion due to the commingling of technical and clinical aspects not previously clarified. Our critical review provides a possible hypothesis about how to match a robot with subject's upper limb functional abilities, but also highlights the need of organizing a clinical consensus conference about the robotic therapy.


Effectiveness of antidepressants. Meta-analysis of dose-effect relationships in randomised clinical trials.
Antidepressant drugs are usually prescribed at low doses, possibly to avoid adverse reactions. No comprehensive review has addressed the issue of dose, clinical response and tolerability in a quantitative way. To determine whether high doses of antidepressants are more effective than low doses, and how safety is affected by dose. Trials comparing two or more doses of the same antidepressant were located, and all antidepressants administered were converted to the equivalent dose of imipramine. Generalised estimating equations were used to analyse percentage improvement and adverse event rate according to dose level. Thirty-three studies were identified. The dose level 100-200 mg imipramine equivalents showed an average improvement of 53% by 'intention-to-treat'. Higher doses were not accompanied by increased efficacy, while lower doses showed reduction in efficacy. Adverse events significantly increased with dose. With a low dose of antidepressants, clinicians trade off a slightly reduced chance of improvement for a higher chance of avoiding adverse reactions.


Intratumoral delivery of bortezomib: impact on survival in an intracranial glioma tumor model.
OBJECTIVE Glioblastoma (GBM) is the most prevalent and the most aggressive of primary brain tumors. There is currently no effective treatment for this tumor. The proteasome inhibitor bortezomib is effective for a variety of tumors, but not for GBM. The authors' goal was to demonstrate that bortezomib can be effective in the orthotopic GBM murine model if the appropriate method of drug delivery is used. In this study the Alzet mini-osmotic pump was used to bring the drug directly to the tumor in the brain, circumventing the blood-brain barrier; thus making bortezomib an effective treatment for GBM. METHODS The 2 human glioma cell lines, U87 and U251, were labeled with luciferase and used in the subcutaneous and intracranial in vivo tumor models. Glioma cells were implanted subcutaneously into the right flank, or intracranially into the frontal cortex of athymic nude mice. Mice bearing intracranial glioma tumors were implanted with an Alzet mini-osmotic pump containing different doses of bortezomib. The Alzet pumps were introduced directly into the tumor bed in the brain. Survival was documented for mice with intracranial tumors. RESULTS Glioma cells were sensitive to bortezomib at nanomolar quantities in vitro. In the subcutaneous in vivo xenograft tumor model, bortezomib given intravenously was effective in reducing tumor progression. However, in the intracranial glioma model, bortezomib given systemically did not affect survival. By sharp contrast, animals treated with bortezomib intracranially at the tumor site exhibited significantly increased survival. CONCLUSIONS Bypassing the blood-brain barrier by using the osmotic pump resulted in an increase in the efficacy of bortezomib for the treatment of intracranial tumors. Thus, the intratumoral administration of bortezomib into the cranial cavity is an effective approach for glioma therapy.


What Can Epidemiological Studies Tell Us about the Impact of Chemical Mixtures on Human Health?
Humans are exposed to a large number of environmental chemicals: Some of these may be toxic, and many others have unknown or poorly characterized health effects. There is intense interest in determining the impact of exposure to environmental chemical mixtures on human health. As the study of mixtures continues to evolve in the field of environmental epidemiology, it is imperative that we understand the methodologic challenges of this research and the types of questions we can address using epidemiological data. In this article, we summarize some of the unique challenges in exposure assessment, statistical methods, and methodology that epidemiologists face in addressing chemical mixtures. We propose three broad questions that epidemiological studies can address: a) What are the potential health impacts of individual chemical agents? b) What is the interaction among agents? And c) what are the health effects of cumulative exposure to multiple agents? As the field of mixtures research grows, we can use these three questions as a basis for defining our research questions and for developing methods that will help us better understand the effect of chemical exposures on human disease and well-being.


Chronic leptin treatment inhibits liver mitochondrial alpha-glycerol-beta-phosphate dehydrogenase in euthyroid rats.
Leptin modulates the hypothalamus-pituitary-thyroid axis and peripheral metabolism of thyroid hormones (THs). We have studied the effect of acute and chronic leptin treatment upon liver mitochondrial glycerol phosphate dehydrogenase activity (mGPD), whose expression and activity are TH dependent. We performed 2 experiments: 1) acute leptin treatment - LepA: adult rats received a single leptin injection (8 microg/100 g BW); 2) chronic leptin treatment - LepC: adult rats received leptin (8 microg/100 g BW) daily, for 6 days. In both experiments, control groups were saline-treated. All rats were sacrificed 2 hours after the last dose. Liver mGPD activity was determined by colorimetric method. Liver D1 activity was measured by the release of (125)I from (125)I-rT3. Serum hormones were measured by RIA. LepA rats showed higher serum thyroid stimulating hormone (TSH) (+ 64%, p<0.05), free T4 (+ 34%, p<0.05), free T3 (+ 64%, p<0.05), and liver D1 activity (+ 85%, p<0.05), but no change in mGPD activity. Since THs increase mGPD activity, the unchanged level in the acute experiment is suggestive of an inhibitory role of leptin. LepC rats presented lower mGPD activity (-1.7-fold, p<0.05) and higher liver D1 activity (+ 32%, p<0.05), but no alteration in serum TSH and free THs. Our results show that leptin downregulates mGPD activity, mainly when hyperleptinemia is chronic.


Simultaneous two-color snapshot view on ultrafast charge and spin dynamics in a Fe-Cu-Ni tri-layer.
Ultrafast phenomena on a femtosecond timescale are commonly examined by pump-probe experiments. This implies multiple measurements, where the sample under investigation is pumped with a short light pulse and then probed with a second pulse at various time delays to follow its dynamics. Recently, the principle of streaking extreme ultraviolet (XUV) pulses in the temporal domain has enabled recording the dynamics of a system within a single pulse. However, separate pump-probe experiments at different absorption edges still lack a unified timing, when comparing the dynamics in complex systems. Here, we report on an experiment using a dedicated optical element and the two-color emission of the FERMI XUV free-electron laser to follow the charge and spin dynamics in composite materials at two distinct absorption edges, simultaneously. The sample, consisting of ferromagnetic Fe and Ni layers, separated by a Cu layer, is pumped by an infrared laser and probed by a two-color XUV pulse with photon energies tuned to the M-shell resonances of these two transition metals. The experimental geometry intrinsically avoids any timing uncertainty between the two elements and unambiguously reveals an approximately 100 fs delay of the magnetic response with respect to the electronic excitation for both Fe and Ni. This delay shows that the electronic and spin degrees of freedom are decoupled during the demagnetization process. We furthermore observe that the electronic dynamics of Ni and Fe show pronounced differences when probed at their resonance, while the demagnetization dynamics are similar. These observations underline the importance of simultaneous investigation of the temporal response of both charge and spin in multi-component materials. In a more general scenario, the experimental approach can be extended to continuous energy ranges, promising the development of jitter-free transient absorption spectroscopy in the XUV and soft X-ray regimes.


Ultrafast relaxation dynamics of phosphine-protected, rod-shaped Au20 clusters: interplay between solvation and surface trapping.
The exact interaction between Au cores and surface ligands remains largely unknown because of the complexity of the structure and chemistry of ligand/Au-core interfaces in ligand-protected Au nanoclusters (AuNCs), which are commonly found in many organic-inorganic complexes. Here, femtosecond transient absorption measurement of the excited-state dynamics of a newly synthesized phosphine-protected cluster [Au20(PPhpy2)10Cl4]Cl2 (1) is reported. Intramolecular charge transfer (ICT) from the Au core to the peripheral ligands was identified. Furthermore, we found that solvation strongly affected ICT at ligand/Au-core interfaces while by choosing several typical alcoholic solvents with different intrinsic solvation times, we successfully observed that excited-state relaxation dynamics together with displacive excited coherent oscillation of Au20 clusters were significantly modulated through the competition between solvation and surface trapping. The results provide a fundamental understanding of the structure-property relationships of the solvation-dependent core-shell interaction of AuNCs for the potential applications in catalysis, sensing and nanoelectronics.


Underuse of pregnancy testing for women prescribed teratogenic medications in the emergency department.
The objectives were to estimate the frequency of pregnancy testing in emergency department (ED) visits by reproductive-aged women administered or prescribed teratogenic medications (Food and Drug Administration categories D or X) and to determine factors associated with nonreceipt of a pregnancy test. This was a retrospective cross-sectional study using 2005 through 2009 National Hospital Ambulatory Medical Care Survey data of ED visits by females ages 14 to 40 years. The number of visits was estimated where teratogenic medications were administered or prescribed and pregnancy testing was not conducted. The association of demographic and clinical factors with nonreceipt of pregnancy testing was assessed using multivariable logistic regression. Of 39,859 sampled visits, representing an estimated 141.0 million ED visits by reproductive-aged females nationwide, 10.1 million (95% confidence interval [CI] = 8.9 to 11.3 million) estimated visits were associated with administration or prescription of teratogenic medications. Of these, 22.0% (95% CI = 19.8% to 24.2%) underwent pregnancy testing. The most frequent teratogenic medications administered without pregnancy testing were benzodiazepines (52.2%; 95% CI = 31.1% to 72.7%), antibiotics (10.7%; 95% CI = 5.0% to 16.3%), and antiepileptics (7.7%; 95% CI = 0.12% to 15.5%). The most common diagnoses associated with teratogenic drug prescription without pregnancy testing were psychiatric (16.1%; 95% CI = 13.6% to 18.6%), musculoskeletal (12.7%; 95% CI = 10.8% to 14.5%), and cardiac (9.5%; 95% CI = 7.6% to 11.3%). In multivariable analyses, visits by older (adjusted odds ratio [AOR] = 0.57, 95% CI = 0.42 to 0.79), non-Hispanic white females (AOR = 0.71; 95% CI = 0.54 to 0.93); visits in the Northeast region (AOR = 0.60; 95% CI = 0.42 to 0.86); and visits during which teratogenic medications were administered in the ED only (AOR = 0.74; 95% CI = 0.57 to 0.97) compared to prescribed at discharge only were less likely to have pregnancy testing. A minority of ED visits by reproductive-aged women included pregnancy testing when patients were prescribed category D or X medications. Interventions are needed to ensure that pregnancy testing occurs before women are prescribed potentially teratogenic medications, as a preventable cause of infant morbidity.


Valproic acid ameliorates C. elegans dopaminergic neurodegeneration with implications for ERK-MAPK signaling.
Parkinson's disease (PD) is a currently incurable neurodegenerative disorder that affects the aging population. The loss of dopaminergic neurons in the substantia nigra is one of the pathological features of PD. The precise causes of PD remain unresolved but evidence supports both environmental and genetic contributions. Current efforts for the treatment of PD are directed toward the discovery of compounds that show promise in impeding age-dependent neurodegeneration in PD patients. Alpha-synuclein (α-Syn) is a human protein that is mutated in specific populations of patients with familial PD. Overexpression of α-Syn in animal models of PD replicates key symptoms of PD, including neurodegeneration. Here, we use the nematode Caenorhabditis elegans as a model system, whereby α-Syn toxicity causes dopaminergic neurodegeneration, to test the capacity of valproic acid (VA) to protect neurons. The results of our study showed that treatment of nematodes with moderate concentrations of VA significantly protects dopaminergic neurons against α-Syn toxicity. Consistent with previously established knowledge related to the mechanistic action of VA in the cell, we showed through genetic analysis that the neuroprotection conferred by VA is inhibited by cell-specific depletion of the C. elegans ortholog of the MAP extracellular signal-regulated kinase (ERK), MPK-1, in the dopaminergic neurons. These findings suggest that VA may exert its neuroprotective effect via ERK-MAPK, or alternately could act with MAPK signaling to additively provide dopaminergic neuroprotection.


Changes in photosynthetic rates and growth following root treatments of tomato plants with phytohormones.
The effects of root applications of kinetin, gibberellic acid (GA3) and indoleacetic acid (IAA) on photosynthesis was measured using an open infrared CO2 gas-exchange system. There was a 30-35% increase in the photosynthetic rates (mg CO2/dm(2)/hr) of attached leaves within 8 hr following root treatment with 0.47 μM kinetin. On a short-term basis (up to 2 days) 0.47 μM kinetin was shown to have the optimal stimulatory effect on photosynthesis, relative growth rate (RGR) and total plant dry weight. If the roots were in contact with 0.47 μM kinetin for longer than two days there was severe branching of the root system and growth was severely decreased. When plants were left in contact with the kinetin treatment for up to 7 days the optimal stimulatory concentration was considerably lower (0.0047 μM) . Plants receiving a 4, 8, or 12 hr pulse with 0.47 μM kinetin to the roots exhibited higher leaf photosynthetic rates than the control. Plants receiving an 8 or 12 hr pulse with 0.47 μM kinetin maintained photosynthetic rates higher than the control for the duration of the experiment (8 days) while the 4 hr pulse remained higher than the control for only 5 days. A sharp decrease in the photosynthetic rate, RGR and total plant dry weight was observed two days following continual treatments with 0.47 μM kinetin to the roots. At low light levels there was approximately a 100% increase in the photosynthetic rate two days following treatment with 0.47 μM kinetin while at a saturating irradiance there was a 30 to 35% increase. Indoleacetic acid either showed no effect on the photosynthetic rate, RGR and total plant dry weight or an inhibitory effect was observed. Either GA3 or kinetin alone were shown to stimulate photosynthesis, RGR and total plant dry weight, however, when GA3 at a 1.4 μM concentration was applied in combination with kinetin at a 0.0047 μM concentration to the roots of tomato plants there was no additive effect. In all cases kinetin dramatically reduced leaf resistance whereas GA3 had no effect.By supplying either GA3 or kinetin to the roots of tomato plants a highly reproducible stimulation in the photosynthetic rate, RGR and total plant dry weight can be achieved at physiologically relevant concentrations, whereas IAA appears to have an inhibitory effect.


A reduction in errors is associated with prospectively recording them.
Error recording and monitoring is an important component of error prevention and quality assurance in the health sector given the huge impact of medical errors on the well-being of patients and the financial loss incurred by health institutions. With this in mind, assessing the effect of reporting errors should be a cause worth pursuing. The object in this study was to examine the null hypothesis that recording and publishing errors do not affect error patterns in a clinical practice. Intraoperative errors and their characteristics were prospectively recorded between May 2000 and May 2013 in the neurosurgical practice of the senior author (M.B.). The error pattern observed between May 2000 and August 2006, which has been previously described (Group A), was compared with the error pattern observed between September 2006 and May 2013 (Group B). A total of 1108 cases in Group A and 974 cases in Group B were surgically treated. A total of 2684 errors were recorded in Group A, while 1892 errors were recorded in Group B. The ratios of cranial to spinal procedures performed in Groups A and B were 3:1 and 10:1, respectively, while the ratios of general to local anesthesia in the two groups were 2:1 and 1.3:1, respectively (p < 0.0001 for both). There was a significantly decreased proportion of cases with error (87% to 83%, p < 0.006), mean errors per case (2.4 to 1.9, p < 0.0001), proportion of error-related complications (16.7% to 5.5%, p < 0.002), and clinical impacts of error (2.7% to 1.0%, p < 0.0001) in Group B compared with Group A. Errors in Group B tended to be more preventable than those in Group A (85.8% vs 78.5%, p < 0.0001). A significant reduction was also noticed with most types of error. A descending trend in the mean errors per case was demonstrated from the years 2001 to 2012; however, an increased severity of errors (22.6% to 29.5%, p < 0.0001) was recorded in Group B compared with Group A. Data in this study showed that the act of recording errors might alter behaviors, resulting in fewer errors.


Visualization of Time-Dependent Distribution of Rifampicin in Rat Brain Using MALDI MSI and Quantitative LCMS/MS.
Rifampicin (RIF) is a major component for short-course chemotherapy against tuberculosis, since it is active against rapidly metabolizing as well as dormant bacteria. According to the Lipinski rules, RIF should not enter the blood-brain barrier. Visualization of tissue drug distribution is of major importance in pharmacological studies; thus, far imaging of RIF in the brain has been limited to positron emission tomography. We propose using matrix-assisted laser desorption/ionization mass spectrometry imaging techniques as a suitable alternative for the visualization and localization of drug tissue distribution. Using the liquid chromatography mass spectrometric (LCMS) technique, we were able to quantify the concentrations of RIF in the uninfected rat brain; we paired this with matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) to show the time-dependent manner in which RIF is able to enter the brain. Our results show that even at the minute concentrations measured with LCMS/MS we were able visualize the drug and show its exact distribution in the rat brain. Other available methods require nuclear labeling and the detection of gamma rays produced by labeled compounds to visualize the compound and its localization; MALDI MSI is a more recently developed technique, which can provide detailed information on drug distribution in tissues when compared to other imaging techniques. This study shows that without any requirement for complex preprocessing we are able to produce images with a relatively improved resolution and localization than those acquired using more complex imaging methods, showing MALDI MSI to be an invaluable tool in drug distribution studies.


[Pharmacokinetics of acebutolol].
Pharmacodynamic data correlating beta-adrenoceptor blockade with plasma level of drug were obtained in healthy male volunteers (3). Direct comparison of the inhibition of isoprenaline induced tachycardia was achieved in each volunteer after the administration of single doses of either acebutolol (300 mg), practolol (400 mg) or propranolol (40 mg). These drugs were approximately equipotent at these doses, at the times of maximum beta-adrenoceptor blockade. Pharmacokinetic data were obtained in hypertensive male patients (4) after treatment with 14C-radioactively labelled acebutolol hydrochloride. Both 'total-14C' levels and specific '14C-acebutol' levels were determined in plasma, urine and faeces. It was shown, by calculation from the renal clearance, that the late biological half-life for the decline of 14C-acebutolol in plasma was 9.4 h and 13.2 h, respectively, in two of these patients treated with a single oral dose (200 mg) of 14C-acebutolol hydrochloride. In one of the patients treated by intravenous infusion (20 mg/10 mn), the late biological half-life plasma was calculated to be 7.5 h. Renal clearance of acebutolol was shown to be close to a mean of 83 ml plasma/mn for each of three patients (two oral and one intravenous) in spite of the fact that one of the orally treated patients had an elevated level of urea in his plasma (47 mg/100 ml) indicative of some impairment of kidney function. The recovery of 14C-radioactivity in the urine (29 p. cent) and faeces (64 p. cent) was 93 p.cent of the dose of labelled acebutolol in one of the orally treated patients. The overall proportion of the dose excreted as unchanged 14C-acebutolol was 62 p.cent. The major metabolite was the product formed by shortening of the butyramido-group of acebutolol to form an acetamido-group. This metabolite was also readily excreted in both urine and faeces and was also detected in an extract of the 4 h plasma from an orally treated patient. It was identified by co-chromatography as the acetyl analogue (M & B 16 942) of acebutolol. It would be detected by the colorimetric assay of acebutolol in plasma because the same aromatic amino-compound (M & B 17 127) would be formed during the acid hydrolysis procedure. A small quantity of an other unidentified metabolite was detected in an extract of freeze-dried urine after autoradiography of a two dimensional thin layer silica-gel chromatogram.


Maternal nutritional risk factors for small for gestational age babies in a developed country: a case-control study.
To assess the effect of maternal diet during pregnancy on the risk of delivering a baby who is small for gestational age (SGA). Case-control study of 844 cases (SGA) and 870 controls (appropriate size for gestational age (AGA)). Only term (37+ completed weeks of gestation) infants were included. Retrospective food frequency questionnaires were completed at birth on the diet at the time of conception and in the last month of pregnancy. At the time of conception, mothers of AGA infants ate significantly more servings of carbohydrate rich food and fruit, and were more likely to have taken folate and vitamin supplements than mothers of SGA infants. There was some evidence that mothers of AGA infants also ate more servings of dairy products, meat, and fish (0.05 < p < 0.1). However, after adjustment for maternal ethnicity, smoking, height, weight, hypertension, and occupation, fish intake (p = 0.04), carbohydrate-rich foods (p = 0.04), and folate supplementation (p = 0.02) were associated with a reduced risk of SGA. In the last month of pregnancy, only iron supplementation was associated with a reduced risk of SGA (p = 0.05) after adjustment for potential confounders. This study suggests that small variations in maternal diets within the normal range during pregnancy in developed countries are associated with differences in birth weight.


[Neuroendocrine regulation under maximal and subjectively optimal physical loads].
The responses of neuroendocrine system to maximum and subjective-optimum physical loading were found to be similar in young men and characterized by activation of adrenocortical, somatotropic and thyroid functions, particularly under maximum loading. The subjective-optimum loading was dosed out by the subjects themselves, differentiated individually and averaged 62% of the maximum oxygen consumption. The parameters of subjective-optimum physical loading were close to the threshold level necessary to stimulate neuroendocrine system and ensure an increase of somatotropine, cortisole, thyroxine and aldosterone concentration in the blood.


Enhanced response of plasma prolactin to metoclopramide during chronic converting enzyme inhibition.
The effect of chronic converting enzyme inhibition with enalapril on the PRA, PRL and plasma aldosterone responses to metoclopramide was studied in 10 patients with mild to moderate essential hypertension. Enalapril reduced supine blood pressure and increased heart rate significantly. PRA and urinary sodium excretion rose significantly. PRA levels did not change after metoclopramide neither during placebo nor during enalapril. The aldosterone response to metoclopramide was not altered by enalapril, indicating that this response is independent of the renin-angiotensin system. The PRL response to metoclopramide was considerably enhanced after 4 weeks of treatment with enalapril. It is proposed that enalapril, by decreasing the formation of angiotensin II, increases the prolactin reserve.


[Current hypotheses on the mechanism of action of lithium].
This article aims to clarify and organize the recent advances in research on the central action mechanism of lithium. It begins by tackling the process of cellular transport of the cation, then it moves on to describe the effects of lithium on ionic distribution and on membranous functioning in general. Neurotransmission in this article is the object of a more detailed study. The metabolism of neurotransmitters (and their disturbances) is first discussed. The emphasis is placed on certain current domains, the gabaergic system in particular. Neurotransmitters, by themselves, do not satisfy any longer the curiosity of biologists. Technological progress has given these scientists access to the intimate functioning of receptors and to their natural support system, the membrane. The stabilizing effect of lithium is mentioned often. Initially seen at a superficial level, it leads to a study of the mechanisms of the transference of information from the receptor to the cell. The membranous phospholipids and the enzymatic systems controlling the metabolism of the second messengers (cyclic nucleotides but also calcium) will provide as many potential targets to the action of lithium.


A genetic screen for sleep and circadian mutants reveals mechanisms underlying regulation of sleep in Drosophila.
In order to characterize the genetic mechanisms underlying sleep, we have carried out a large-scale screen in Drosophila to identify short-sleeping mutants. The objectives of this study were as follows: (1) characterize the phenotypes of the shortest-sleeping mutants; (2) examine whether changes in arousal threshold or sleep homeostasis underlie short-sleeping phenotypes; (3) clone a gene affected in one of the shortest sleepers; and (4) investigate whether circadian mutants can be identified using light:dark (L:D) locomotor data obtained for studying sleep behavior. Locomotor activity was measured using the Drosophila Activity Monitoring System in a 12:12 L:D cycle. Drosophila research laboratory. Adult flies from the 2nd chromosome Zuker collection, which contain mutations in most of the nonessential genes on the Drosophila 2nd chromosome. Our analysis of sleep characteristics suggests that daily activity (but not waking activity) correlates with daily sleep time and that defects in sleep maintenance are more common than defects in sleep initiation. Our shortest sleepers have intact or increased sleep rebound following sleep deprivation but show reduced thresholds for arousal. Molecular analysis of one of the short-sleeping lines indicates that it is a novel allele of a dopamine transporter (DAT). Finally, we describe a novel approach for identifying circadian mutants using L:D data. Our data suggest that most short-sleeping mutant phenotypes in Drosophila are characterized by an inability to stay asleep, most likely because of a reduced arousal threshold.


Evidence for the Role of Blue Light in the Development of Uveal Melanoma.
Uveal melanoma is the most common malignancy of the adult eye. Although it is a relatively infrequent tumor, clinical prognosis is often poor owing to a high incidence of aggressive metastatic disease, for which there are limited treatment options. Little is known about the etiology of this condition, although several risk factors have been identified. Unlike cutaneous melanoma, however, ultraviolet radiation does not figure prominently among these risk factors. In this review, we focus on an associated form of visible electromagnetic radiation, high-energy short-wave (blue) light, a causative agent in various forms of age-related retina damage, as a previously overlooked risk factor in uveal melanoma development and progression. Finally, we discuss the impact of these data on contemporary ocular therapy, particularly the debate surrounding the filtering capabilities of intraocular lenses used to replace dysfunctional crystalline lenses during cataract surgery.


[Somatoparaphrenia in a patient with schizophrenia and addiction problems: 'That arm is not mine, doctor'].
Somatoparaphrenia is a disorder of body perception, usually on the left side. One or both limbs are seen as foreign or as belonging to somebody else. In the literature this rare phenomenon has been described in patients with brain damage, usually due to an infarction or other lesion to the right parietal lobe. We describe a patient with schizophrenia and addiction problems who believed that his left forearm was not his, but rather belonged to a Spanish girl. An EEG and an MRI of the brain showed no abnormalities. Despite years of antipsychotic treatment, the delusion persisted. To rule out neurological causes we recommend auxiliary investigations in all patients with somatoparaphrenia. No evidence-based treatments are known for this monothematic delusion in the context of schizophrenia.


Dysphagia lusoria: aberrant right subclavian artery with a Kommerell's diverticulum.
A retroesophageal right subclavian artery, the most common congenital aortic arch abnormality, is an unusual cause of dysphagia in adults. The embryologic abnormality of the aortic arch is involution of the fourth vascular arch, along with the right dorsal aorta, leaving the seventh intersegmental artery attached to the descending aorta. This persistent intersegmental artery assumes a retroesophageal position as it proceeds out of the thorax into the arm. Since compression of the esophagus by this right subclavian artery may produce dysphagia, the term "dysphagia lusoria" ("dysphagia by freak of nature"), has been used to describe the symptom complex. The presence of an aneurysm of the artery or Kommerell's diverticulum at its aortic origin is more likely to produce symptoms from esophageal compression. This case presents a middle aged adult with an associated Kommerell's diverticulum and dysphagia. Surgical correction was used to relieve his symptoms and to correct the diverticulum of the proximal right subclavian artery. The embryologic changes that occur are discussed in detail.


Auxiliary Proteins are the Predominant Determinants of Differential Efficacy of Clinical Candidates Acting as AMPA Receptor Positive Allosteric Modulators.
Positive allosteric modulators (PAMs) of AMPA receptors boost cognitive performance in preclinical and clinical studies. Their therapeutic window is narrow, however, and clinical application will likely only occur if greater discrimination in activity is achieved. Toward that end, we compared the modulatory activity of two PAMs recently considered as clinical candidates, LY451395 (mibampator) and PF-04958242/BIIB104, on recombinant and native AMPA receptors (AMPARs). We found that the principle molecular determinant that shaped modulatory activity of both PAMs on deactivation (recombinant) and decay (synaptic) of AMPARs was the auxiliary protein incorporated into the receptor complexes. AMPARs containing the stargazin/γ2 transmembrane AMPAR regulatory protein (TARP) were slowed to a >10-fold degree by both PAMs as compared with those incorporating γ8 TARP. Neither subunit composition nor flip/flop splice variation had substantive effect. Similarly, stargazin/γ2-containing mossy fiber EPSCs in cerebellar granule neurons were slowed to a ∼5-fold greater degree than EPSCs in hippocampal CA1 pyramidal cell neurons, which express the γ8 TARP. LY451395 exhibited greater efficacy than BIIB104 at both synapses. These studies provide insight into the receptor constituents that determine efficacy of sulfonamide PAMs. We conclude that compounds that discriminate between AMPARs complexed with distinct TARPs, and particularly those with lower stargazin/γ2 efficacy such as BIIB104, could act as viable procognitive therapeutics. SIGNIFICANCE STATEMENT: Positive allosteric modulators (PAMs) of AMPA receptors enhance cognitive function in a variety of preclinical models. A clearer understanding of the critical determinants of PAM activity could yield critical insight into pathways to maximize their therapeutic index. Here we show that auxiliary proteins for AMPARs play a major, but thus far underappreciated, role in shaping recombinant and neuronal AMPAR modulation by two clinical candidate PAMs. These data will inform both clinical outcomes as well as future rational development of new modulators.


Ubiquitin-Mediated Proteasomal Degradation of Oleosins is Involved in Oil Body Mobilization During Post-Germinative Seedling Growth in Arabidopsis.
In oleaginous seeds, lipids--stored in organelles called oil bodies (OBs)--are degraded post-germinatively to provide carbon and energy for seedling growth. To date, little is known about how OB coat proteins, known as oleosins, control OB dynamics during seed germination. Here, we demonstrated that the sequential proteolysis of the five Arabidopsis thaliana oleosins OLE1-OLE5 begins just prior to lipid degradation. Several post-translational modifications (e.g. phosphorylation and ubiquination) of oleosins were concomitant with oleosin degradation. Phosphorylation occurred only on the minor OLE5 and on an 8 kDa proteolytic fragment of OLE2. A combination of immunochemical and proteomic approaches revealed ubiquitination of the four oleosins OLE1-OLE4 at the onset of OB mobilization. Ubiquitination topology was surprisingly complex. OLE1 and OLE2 were modified by three distinct and predominantly exclusive motifs: monoubiquitin, K48-linked diubiquitin (K48Ub(2)) and K63-linked diubiquitin. Ubiquitinated oleosins may be channeled towards specific degradation pathways according to ubiquitination type. One of these pathways was identified as the ubiquitin-proteasome pathway. A proteasome inhibitor (MG132) reduced oleosin degradation and induced cytosolic accumulation of K48Ub(2)-oleosin aggregates. These results indicate that K48Ub(2)-modified oleosins are selectively extracted from OB coat and degraded by the proteasome. Proteasome inhibition also reduced lipid hydrolysis, providing in vivo evidence that oleosin degradation is required for lipid mobilization.


Racial differences in time to treatment for melanoma.
Longer time from diagnosis to definitive surgery (TTDS) is associated with increased melanoma-specific mortality. Although black patients present with later-stage melanoma and have worse survival than non-Hispanic white patients, the association between race and TTDS is unknown. To investigate racial differences in time to melanoma treatment. Retrospective review of the National Cancer Database (2004-2015). Multivariable logistic regression was used to evaluate the association of race with TTDS, controlling for sociodemographic/disease characteristics. Of the 233,982 patients with melanoma identified, 1221 (0.52%) were black. Black patients had longer TTDS for stage I to III melanoma (P < .001) and time to immunotherapy (P = .01), but not for TTDS for stage IV melanoma or time to chemotherapy (P > .05 for both). When sociodemographic characteristics were controlled for, black patients had over twice the odds of having a TTDS between 41 and 60 days, over 3 times the odds of having a TTDS between 61 and 90 days, and over 5 times the odds of having a TTDS over 90 days. Racial differences in TTDS persisted within each insurance type. Patients with Medicaid had the longest TTDS (mean, 60.4 days), and those with private insurance had the shortest TTDS (mean, 44.6 days; P < .001 for both). Targeted approaches to improve TTDS for black patients are integral in reducing racial disparities in melanoma outcomes.


Speeding up tandem mass spectrometry based database searching by peptide and spectrum indexing.
Database searching is the technique of choice for shotgun proteomics, and to date much research effort has been spent on improving its effectiveness. However, database searching faces a serious challenge of efficiency, considering the large numbers of mass spectra and the ever fast increase in peptide databases resulting from genome translations, enzymatic digestions, and post-translational modifications. In this study, we conducted systematic research on speeding up database search engines for protein identification and illustrate the key points with the specific design of the pFind 2.1 search engine as a running example. Firstly, by constructing peptide indexes, pFind achieves a speedup of two to three compared with that without peptide indexes. Secondly, by constructing indexes for observed precursor and fragment ions, pFind achieves another speedup of two. As a result, pFind compares very favorably with predominant search engines such as Mascot, SEQUEST and X!Tandem.