Randomized Controlled Trial of the Gastrin/CCK2 Receptor Antagonist Netazepide in Patients with Barrett's Esophagus.
Hypergastrinemia has been associated with high-grade dysplasia and adenocarcinoma in patients with Barrett's esophagus, and experimental studies suggest proinflammatory and proneoplastic effects of gastrin on Barrett's esophagus. This is of potential concern, as patients with Barrett's esophagus are treated with medications that suppress gastric acid production, resulting in increased physiologic levels of gastrin. We aimed to determine whether treatment with the novel gastrin/CCK2 receptor antagonist netazepide reduces expression of markers associated with inflammation and neoplasia in Barrett's esophagus. This was a randomized, double-blind, placebo-controlled trial of netazepide in patients with Barrett's esophagus without dysplasia. Subjects were treated for 12 weeks, with endoscopic assessment at baseline and at end of treatment. The primary outcome was within-individual change in cellular proliferation as assessed by Ki67. Secondary analyses included changes in gene expression, assessed by RNA-sequencing, and safety and tolerability. A total of 20 subjects completed the study and were included in the analyses. There was no difference between arms in mean change in cellular proliferation (netazepide: +35.6 Ki67+ cells/mm2, SD 620.7; placebo: +307.8 Ki67+ cells/mm2, SD 640.3; P = 0.35). Netazepide treatment resulted in increased expression of genes related to gastric phenotype (TFF2, MUC5B) and certain cancer-associated markers (REG3A, PAX9, MUC1), and decreased expression of intestinal markers MUC2, FABP1, FABP2, and CDX1 No serious adverse events related to study drug occurred. The gastrin/CCK2 receptor antagonist netazepide did not reduce cellular proliferation in patients with nondysplastic Barrett's esophagus. Further research should focus on the biological effects of gastrin in Barrett's esophagus.Prevention Relevance: Treatment of patients with Barrett's esophagus with a gastrin/CCK2 receptor antagonist did not have obvious chemopreventive effects.


Surgical treatment of invasive squamous cell carcinoma of the penis: retrospective analysis of 350 cases.
Between 1960 and 1987, 414 patients with invasive squamous cell carcinoma of the penis were referred to the Brazilian National Cancer Institute. Inguinal metastases were demonstrated by lymphadenectomy in 39% of the 23 patients with stage N0, 49% of 92 with stages N1 and N2, and 100% of 18 with stage N3 disease. We analyzed the followup of 350 patients who underwent surgical treatment. In 224 patients (64%) amputation or some form of penile surgery was done initially, while 102 (29%) underwent amputation and lymphadenectomy, and 24 (7%) underwent palliative surgery for advanced squamous cell carcinoma. The statistics revealed a better 5-year survival rate for the patients who underwent lymphadenectomy concomitantly with penile surgery compared to those who underwent delayed lymphadenectomy (p < 0.001). Patients in whom systematic lymphadenectomy was negative had a better prognosis than those with positive systematic lymphadenectomy results (p < 0.001). The latter patients had a better prognosis compared with those in whom delayed lymphadenectomy was positive (p = 0.0103). Patients with well and moderately differentiated carcinoma had a higher survival rate at 5 years than did those with poorly differentiated carcinoma (p < 0.001 and p = 0.003, respectively). All deaths from metastatic disease occurred within 24 months among the patients who underwent systematic lymphadenectomy and within 5 years after simple penile surgery. In the short term, surgical debulking combined with reconstruction techniques allowed for improved quality of life in patients with advanced local-regional disease.


Management of acute musculoskeletal pain (excluding low back pain): protocol for a systematic review and network meta-analysis of randomised trials.
Acute, non-low back-related musculoskeletal pain is common and associated with significant socioeconomic costs. No review has evaluated all interventional studies for acute musculoskeletal pain, which limits attempts to make inferences regarding the relative effectiveness of treatments. We will conduct a systematic review of all randomised controlled trials evaluating therapies for acute musculoskeletal pain (excluding low back pain). We will identify eligible, English-language, trials by a systematic search of the Cumulative Index to Nursing and Allied Health Literature (CINAHL), Embase, Medline, Physiotherapy Evidence Database (PEDro) and the Cochrane Central Register of Controlled Trials (CENTRAL) from inception to February 2018. Eligible trials will: (1) enrol patients presenting with acute, non-low back-related musculoskeletal pain (duration of pain ≤4 weeks), and (2) randomise patients to alternative interventions or an intervention and a placebo/sham arm. Fractures will be considered ineligible, unless they are non-surgical and therapy is directed at pain relief. Pairs of reviewers will, independently and in duplicate, screen titles and abstracts of identified citations, review the full texts of potentially eligible trials and extract information from eligible trials. We will use a modified Cochrane instrument to evaluate risk of bias. Disagreements will be resolved through discussion to achieve consensus. We will use the Grading of Recommendations Assessment, Development and Evaluation approach to evaluate the quality of evidence supporting treatment effects. When possible, we will conduct: (1) in direct comparisons, a random-effect meta-analysis to establish the effectiveness of therapeutic interventions on patient-important outcomes; and (2) multiple treatment comparison meta-analysis to assess the relative effects of treatments. We will use a priori hypotheses to explain heterogeneity between studies. We will use STATA V.14.2 for all analyses. No research ethics approval is required for this systematic review, as no confidential patient data will be used. The results of this systematic review will be disseminated through publication in a peer-reviewed journal, conference presentations and will inform a clinical practice guideline. CRD42018094412.


Counselling the patient with prostate cancer about treatment-related erectile dysfunction.
At present, a significant proportion of patients with prostate cancer are diagnosed at an early stage and may receive treatments able to bring about the long-term control of the disease. Thus, the impact of available treatments on the patient's quality of life has been gaining increasing importance; for patients with prostate cancer, counselling on the treatment-related effects on sexual function has become mandatory. Radical prostatectomy is very frequently performed in patients with clinically localized prostate cancer. Postoperative erectile function has been reported as being satisfactory in the majority of the patients operated on in centers of excellence for this procedure. However, overall, the results for postoperative potency are disappointing in view of the large amounts of data available from community practices. Attempts to improve postoperative potency include the intraoperative use of cavernous nerve stimulation and grafting of peripheral nerves to restore the innervation of the corpora cavernosa. Erectile dysfunction has also been associated with prostate radiotherapy. It has been shown that both ultrasound-guided brachytherapy and three-dimensional conformal radiation therapy cause an impairment of erectile function that is usually seen some time after the completion of therapy. Treatment with sildenafil citrate remains a viable option both for patients treated with radical prostatectomy (in whom the cavernous nerve function is at least partially present) and in patients treated with radiotherapy.


Microsatellite primers for the fungi Rhizopogon kretzerae and R. salebrosus (Rhizopogonaceae) from 454 shotgun pyrosequencing.
Rhizopogon kretzerae and R. salebrosus (Rhizopogonaceae) are ectomycorrhizal fungi symbiotic with pines and the mycoheterotrophic plant Pterospora andromedea (Ericaceae). Microsatellite loci will allow population genetic study of fungal hosts to P. andromedea. • Shotgun pyrosequencing of R. kretzerae DNA resulted in primer development of 23 perfect microsatellite loci and screened across two populations each for R. kretzerae and R. salebrosus. Twelve loci were polymorphic in R. kretzerae populations, and 11 loci cross-amplified in R. salebrosus populations. For R. kretzerae and R. salebrosus, number of alleles was one to eight and one to nine, respectively, and observed heterozygosity ranged from 0.00-0.57 and 0.00-0.70, respectively. • These are the first microsatellite loci developed for any species within Rhizopogon subgenus Amylopogon. These microsatellite loci will be used in conservation genetic studies of rare to endangered eastern populations and to compare plant and fungal population genetic structure at different hierarchical levels.


Rupture of the corpus cavernosum of the penis.
Rupture of the corpus cavernosum ("fracture" of the penis) is an uncommon injury that occurs in an erect penis. The tunica albuginea is torn during coitus or as a result of external forces, resulting in sudden pain, swelling, and discoloration of the penis. Urethral rupture is associated with the cavernous injury in approximately 10% of cases. Treatment of the injury aims at full functional and cosmetic recovery. Nonoperative treatment results in a 10% incidence of penile deformity and requires a substantial hospital stay. However, surgical repair eliminates resultant deformity and allows for earlier hospital discharge. Three patients had fractured penises, including one penis associated with a torn urethra. Early surgical repair was performed in each patient, and the results were excellent. Prompt surgical repair should be given to all patients with a fractured penis.


Glucocorticoids and checkpoint tyrosine kinase inhibitors stimulate rat pancreatic beta cell proliferation differentially.
Cell therapy for diabetes could benefit from the identification of small-molecule compounds that increase the number of functional pancreatic beta cells. Using a newly developed screening assay, we previously identified glucocorticoids as potent stimulators of human and rat beta cell proliferation. We now compare the stimulatory action of these steroid hormones to a selection of checkpoint tyrosine kinase inhibitors that were also found to activate the cell cycle-in beta cells and analyzed their respective effects on DNA-synthesis, beta cell numbers and expression of cell cycle regulators. Our data using glucocorticoids in combination with a receptor antagonist, mifepristone, show that 48h exposure is sufficient to allow beta cells to pass the cell cycle restriction point and to become committed to cell division regardless of sustained glucocorticoid-signaling. To reach the end-point of mitosis another 40h is required. Within 14 days glucocorticoids stimulate up to 75% of the cells to undergo mitosis, which indicates that these steroid hormones act as proliferation competence-inducing factors. In contrast, by correlating thymidine-analogue incorporation to changes in absolute cell numbers, we show that the checkpoint kinase inhibitors, as compared to glucocorticoids, stimulate DNA-synthesis only during a short time-window in a minority of cells, insufficient to give a measurable increase of beta cell numbers. Glucocorticoids, but not the kinase inhibitors, were also found to induce changes in the expression of checkpoint regulators. Our data, using checkpoint kinase-specific inhibitors further point to a role for Chk1 and Cdk1 in G1/S transition and progression of beta cells through the cell cycle upon stimulation with glucocorticoids.


Scalable thioarylation of unprotected peptides and biomolecules under Ni/photoredox catalysis.
Site-specific functionalization of unprotected native peptides and biomolecules remains a useful transformation in synthetic design and chemical biology, yet until recently, advancements in transition metal-catalyzed methods, which have prevailed in organic synthesis, have been relatively ineffective when applied to large and structurally complex biomolecules. Here, the mechanistically distinct, Ni/photoredox-catalyzed arylation of unprotected, native thiols (e.g., cysteine residues) is reported - a process initiated through a visible light-promoted, hydrogen atom transfer (HAT) event under ambient conditions. Sub-stoichiometric loadings of the dual-catalyst system (≤5 mol%) are employed, granting excellent site-specificity, broad substrate scope, and low chemical waste. Reaction scalability (from μg to grams) has been achieved through modest reagent adjustments, and high throughput experimentation (HTE) demonstrates the ease of reaction setup, enabling prompt screening of aryl halide coupling partners and conditions. Scores of thiol substrates and aryl entities were examined and effectively conjugated, suggesting further diverse, practical applications.


Exercise in chronic pulmonary disease: limitations and rehabilitation.
Chronic pulmonary disease is common in the community and increasing in prevalence. Although numerous etiologies exist, chronic obstructive pulmonary disease secondary to tobacco smoking, and asthma constitute the majority of cases. The important impact of these diseases on patients is disabling breathlessness and impairment of functional exercise capacity. The symptoms set up a vicious cycle leading to physical deconditioning and worsening exercise performance. The discipline of pulmonary rehabilitation has been conclusively shown to reverse this process, resulting in improved functional capacity and reduced breathlessness. Pulmonary rehabilitation, therefore, should be viewed as essential secondary preventative care for the majority of patients with chronic pulmonary disease. As such, early disease recognition and implementation of exercise reconditioning is important. In order to be maximally effective, pulmonary rehabilitation must recognize the complex underlying pathophysiology in chronic pulmonary disease and be customized to the individual patient. The chosen mode of exercise training should recognize that in order to be truly beneficial, any physiological responses need to translate readily into improvements in activities of daily living. Therefore, sessions in pulmonary rehabilitation should concentrate on exercises that have proven useful in this regard. Aerobic and resistance exercise prescriptions should be rigorous, scientifically based, and derived from an understanding of the basic principles of the human response to exercise prescription. Each of these exercise prescriptions should encompass the basic principles of intensity, frequency, duration, and progression suitably modified for the individual patient with chronic pulmonary disease.


Innate immune recognition of microbial cell wall components and microbial strategies to evade such recognitions.
The innate immune system constitutes the first line of defence against invading microbes. The basis of this defence resides in the recognition of defined structural motifs of the microbes called "Microbial associated molecular patterns" that are absent in the host. Cell wall, the outer layer of both bacterial and fungal cells, a unique structure that is absent in the host and is recognized by the germ line encoded host receptors. Nucleotide oligomerization domain proteins, peptidoglycan recognition proteins and C-type lectins are host receptors that are involved in the recognition of bacterial cell wall (usually called peptidoglycan), whereas fungal cell wall components (N- and O-linked mannans, β-glucans etc.) are recognized by host receptors like C-type lectins (Dectin-1, Dectin-2, mannose receptor, DC-SIGN), Toll like receptors-2 and -4 (TLR-2 and TLR-4). These recognitions lead to activation of a variety of host signaling cascades and ultimate production of anti-microbial compounds including phospholipase A2, antimicrobial peptides, lysozyme, reactive oxygen and nitrogen species. These molecules act in cohort against the invading microbes to eradicate infections. Additionally pathogen recognition leads to the production of cytokines, which further activate the adaptive immune system. Both pathogenic and commensal bacteria and fungus use numerous strategies to subvert the host defence. These strategies include bacterial peptidoglycan glycan backbone modifications by O-acetylation, N-deacetylation, N-glycolylation and stem peptide modifications by amidation of meso-Diaminopimelic acid; fungal cell wall modifications by shielding the β-glucan layer with mannoproteins and α-1,3 glucan. This review focuses on the recent advances in understanding the role of bacterial and fungal cell wall in their innate immune recognition and evasion strategies.


The immunopathology of multiple sclerosis: an overview.
Multiple sclerosis (MS) is traditionally seen as an inflammatory demyelinating disease, characterized by the formation of focal demyelinated plaques in the white matter of the central nervous system. In this review we describe recent evidence that the spectrum of MS pathology is much broader. This includes demyelination in the cortex and deep gray matter nuclei, as well as diffuse injury of the normal-appearing white matter. The mechanisms responsible for the formation of focal lesions in different patients and in different stages of the disease as well as those involved in the induction of diffuse brain damage are complex and heterogeneous. This heterogeneity is reflected by different clinical manifestations of the disease, such as relapsing or progressive MS, and also explains at least in part the relation of MS to other inflammatory demyelinating diseases.


Gut microbiome mediates the protective effects of exercise after myocardial infarction.
Gut microbiota plays important roles in health maintenance and diseases. Physical exercise has been demonstrated to be able to modulate gut microbiota. However, the potential role of gut microbiome in exercise protection to myocardial infarction (MI) remains unclear. Here, we discovered exercise training ameliorated cardiac dysfunction and changed gut microbial richness and community structure post-MI. Moreover, gut microbiota pre-depletion abolished the protective effects of exercise training in MI mice. Furthermore, mice receiving microbiota transplants from exercised MI mice had better cardiac function compared to mice receiving microbiota transplants from non-exercised MI mice. Mechanistically, we analyzed metabolomics in fecal samples from exercised mice post-MI and identified 3-Hydroxyphenylacetic acid (3-HPA) and 4-Hydroxybenzoic acid (4-HBA), which could be applied individually to protect cardiac dysfunction post-MI and apoptosis through NRF2. Together, our study provides new insights into the role of gut microbiome in exercise protection to MI, offers opportunities to modulate cardiovascular diseases by exercise, microbiome and gut microbiota-derived 3-HPA and 4-HBA. Video Abstract.


Model-Based Analysis Reveals a Sustained and Dose-Dependent Acceleration of Wound Healing by VEGF-A mRNA (AZD8601).
Intradermal delivery of AZD8601, an mRNA designed to produce vascular endothelial growth factor A (VEGF-A), has previously been shown to accelerate cutaneous wound healing in a murine diabetic model. Here, we develop population pharmacokinetic and pharmacodynamic models aiming to quantify the effect of AZD8601 injections on the dynamics of wound healing. A dataset of 584 open wound area measurements from 131 mice was integrated from 3 independent studies encompassing different doses, dosing timepoints, and number of doses. Evaluation of several candidate models showed that wound healing acceleration is not likely driven directly by time-dependent VEGF-A concentration. Instead, we found that administration of AZD8601 induced a sustained acceleration of wound healing depending on the accumulated dose, with a dose producing 50% of the maximal effect of 92 µg. Simulations with this model showed that a single dose of 200 µg AZD8601 can reduce the time to reach 50% wound healing by up to 5 days.


Chronic treatment with the phosphodiesterase type 5 inhibitors sildenafil and tadalafil display anxiolytic effects in Flinders Sensitive Line rats.
There are conflicting results from behavioural studies regarding whether the activation or inhibition of the cGMP-nitric oxide (NO) pathway induces anxiolytic-like behaviour. Sildenafil, an inhibitor of cGMP-selective phosphodiesterase-5, increases anxiety acutely, but previous evidence suggests that its chronic administration may be anxiolytic, and could involve a cholinergic interaction. We used the Flinders Sensitive Line (FSL) rat, a genetic model of depression that presents with increased anxiety- and depression-like behaviour, to investigate the action of chronic treatment with the PDE5 inhibitors sildenafil or tadalafil, with/without atropine on social interaction behaviour, a correlate for anxiety. Fluoxetine was used as positive control, with validation performed using Flinders Resistant Line (FRL) rats. In order to relate behavioural changes to brain penetration, we determined the concentration of sildenafil in cortex and hippocampus of rats following the schedule above using LC-MS/MS. FSL rats displayed significantly reduced social interactive behaviour than FRL rats, while sildenafil, tadalafil, and fluoxetine significantly reversed these deficits. Atropine did not exert effects on social interactive behaviour, nor did it modulate the effects of sildenafil or tadalafil. Sildenafil was present in cortex and hippocampus regions in lower nanomolar concentrations after chronic treatment, in agreement with the binding to PDE5 required for pharmacological effects. This study emphasizes the complicated regulation of anxiety by the cGMP-NO system, and provides supporting evidence for an anxiolytic action after the chronic activation of this pathway. As far as we know this is also the first report to formally demonstrate that sildenafil effectively crosses the blood-brain barrier to elicit central effects.


Metachronous Isolated Contralateral Lung Metastasis from Pulmonary Adenosquamous Carcinoma with EGFR Mutation.
Lung metastasis and metachronous double primary lung cancer are both common and often present diagnostic challenges. We present a case of metachronous isolated contralateral lung metastasis from pulmonary adenosquamous carcinoma with EGFR mutation. A 75-yearold woman presented with left lung nodule on a routine follow-up chest radiograph. She had had surgery for pulmonary adenocarcinoma with EGFR Ex21 L858R mutation 6 years ago. She underwent surgical resection, and histologic findings revealed adenosquamous carcinoma with the same EGFR mutation. Re-assessment of the resected specimen of the primary tumor resected 6 years ago revealed the morphologically similarity to the left lung tumor. Based on morphological and genetic identity, final diagnosis was adenosquamous cell carcinoma and metachronous isolated contralateral lung metastasis. The diagnosis of metachronous isolated metastasis is difficult but important for appropriate management and prediction of prognosis. A careful pathological examination and evaluation of genetic abnormality are needed to make the correct diagnosis.


Prenatal group visit program for a population with limited English proficiency.
The declining number of family physicians providing pregnancy care is of concern because they are an important source of pregnancy care in underserved communities. Innovative approaches might reinforce family physician participation in pregnancy care for the underserved. Since group prenatal visits have been shown to improve patient education, support, and satisfaction, we implemented and evaluated a group prenatal visit program for Japanese women in Michigan, an underserved population because of their limited proficiency with English. We conducted a convergent quantitative and qualitative mixed methods evaluation involving repeated survey administration (program evaluations, 4-item Patient Health Questionnaire, pregnancy distress questionnaire) to participants during 5 group visits and in-depth postpartum interviews in the University of Michigan Japanese Family Health Program setting. We conducted independent quantitative and qualitative analytics and then thematically integrated these data. Cultural adaptations to the Centering Pregnancy format involved changes in total visits, educational content, and participation format. Based on 5 groups attending 5 sessions each, 42 women evaluated the program through 158 surveys after the sessions. Participants evaluated multiple parameters positively: being with other pregnant women (98%), improving their understanding about prenatal care (96%), preparation for labor and delivery (96%), organization of visits (94%), and preparation for newborn care (85%). In final evaluations, 96% to 100% of participants rated 7 educational topics as "covered" or "covered well." Qualitative interviews with 20 women revealed positive views of social support from prenatal group visits and group facilitation but mixed enthusiasm for clinical assessments in the prenatal group visit setting and partner and children attendance at the sessions. This research demonstrates the feasibility and cultural acceptability of prenatal group visits for Japanese women. Prenatal group visits provided education and social support for Japanese women during the perinatal and postpartum periods that were not otherwise accessible in Japanese. This study confirms the feasibility of family physicians providing prenatal group visits and extends the literature of the applicability of prenatal group visits for patients with limited English skills.


[Progressively acting antithrombins and plasma heparin resistance in ischemic heart disease].
A comparative study of activity of the progressively acting antithrombin and their heparin-cofactor activity was undertaken in 27 patients with angina pectoris and 46 with macrofocal myocardial infarction. It was established that the count of thrombin inactivation during 5 minutes (Abiligaard method) shows no disorders in the majority of patients. Only in 3 patients with the primary antithrombin III and 3 (40% of patients with acute myocardial infarction showed a marked decrease of this parameter. Inactivation count of the endogenous thrombin according to the "self-coagulogram" during one hour enabled one to show decrease of activity of antithrombin in of 80% of patients. The heparin-cofactor activity in the serial heparin-thrombin test is markedly disrupted, which was used to compute the indices of activity of antithrombin and of the antithrombin plasma reserve. This disorder did not depend much on the increase of the antiheparin factor of thrombocytes in plasma.


Granulomatous synovitis and Crohn's disease.
A case of Crohn's disease of the colon, diagnosed after the discovery of a granulomatous synovitis of the wrist in a 40-year-old woman is described. The clinical and histological features of the articular and extraarticular inflammatory lesions of Crohn's disease are reviewed.


Improving care through a medication administration process action team.
The article discusses the use of a process action team to improve the medication administration system. The interdisciplinary team suggests alternatives after reviewing current practice, systems, and procedures. After suggested alternatives are applied, the medication dispensing and administration system improves and enhances the delivery of medications to patients. This opportunity to improve care explains the use of the process action team in patient care system improvement.


Syntheses of molecularly imprinted polymers: Molecular recognition of cyproheptadine using original print molecules and azatadine as dummy templates.
The use of custom-made polymeric materials with high selectivities as target molecules in solid-phase extraction (SPE), known as molecularly imprinted solid-phase extraction (MISPE), is becoming an increasingly important sample preparation technique. However, the potential risk of leakage of the imprinting molecules during the desorption phase has limited application. The use of a mimicking template, called a dummy molecular imprinting polymer (DMIP), that bears the structure of a related molecule and acts as a putative imprinting molecule may provide a useful solution to this problem. In the current study, cyproheptadine (CPH) and azatadine (AZA) were used as templates in the development of an MIP and DMIP for acrylic acid and methacrylic acid monomers. Our results indicate that DMIPs have equal recognition of CPH, avoiding the problem of leakage of original template during the desorption phase relative to MIPs synthesized in presence of the print molecule CPH. Examination of the surface structure of the two polymer products by SEM shows appreciable differences in structural morphology and function of the monomers employed. These results are well supplemented by data obtained for swelling ratios and solvent uptake. Molecular modelling of CPH and AZA suggests that both substrates are similar in shape and volume.


Multiphoton excitation provides optical sections from deeper within scattering specimens than confocal imaging.
Multiphoton excitation fluorescence imaging generates an optical section of sample by restricting fluorophore excitation to the plane of focus. High photon densities, achieved only in the focal volume of the objective, are sufficient to excite the fluorescent probe molecules by density-dependent, multiphoton excitation processes. We present comparisons of confocal with multiphoton excitation imaging of identical optical sections within a sample. These side-by-side comparisons of imaging modes demonstrate a significant advantage of multiphoton imaging; data can be obtained from deeper within biological specimens. Observations on a variety of biological samples showed that in all cases there was at least a twofold improvement in the imaging penetration depth obtained with multiphoton excitation relative to confocal imaging. The more pronounced degradation in image contrast deep within a confocally imaged sample is primarily due to scattered emission photons, which reduce the signal and increase the local background as measurements of point spread functions indicated that resolution does not significantly change with increasing depth for either mode of microscopy. Multiphoton imaging does not suffer from degradation of signal-to-background to nearly the same extent as confocal imaging because this method is insensitive to scatter of the emitted signal. Direct detection of emitted photons using an external photodetector mounted close to the objective (possible only in a multiphoton imaging system) improves system sensitivity and the utilization of scattered emission photons for imaging. We demonstrate that this technique provides yet further improvements in the capability of multiphoton excitation imaging to produce good quality images from deeper within tissue relative to confocal imaging.


Phase transformation-driven artificial muscle mimics the multifunctionality of avian wing muscle.
Skeletal muscle provides a compact solution for performing multiple tasks under diverse operational conditions, a capability lacking in many current engineered systems. Here, we evaluate if shape memory alloy (SMA) components can serve as artificial muscles with tunable mechanical performance. We experimentally impose cyclic stimuli, electric and mechanical, to an SMA wire and demonstrate that this material can mimic the response of the avian humerotriceps, a skeletal muscle that acts in the dynamic control of wing shapes. We next numerically evaluate the feasibility of using SMA springs as artificial leg muscles for a bipedal walking robot. Altering the phase offset between mechanical and electrical stimuli was sufficient for both synthetic and natural muscle to shift between actuation, braking and spring-like behaviour.


Using human genetics to discover new therapeutic targets for plasma lipids.
Genetic variation arises through multiple different alleles that vary in frequency and severity of effect. Mutations that give rise to Mendelian disorders, such as the LDL receptor (LDLR) mutations that result in familial hypercholesterolaemia, are efficiently winnowed from the population by purifying selection and are almost inevitably rare. Conversely, alleles that are common in the population (such that homozygotes for the minor allele are present even in modest sample sizes) typically have very modest phenotypic effects. Mutations in the gene for proprotein convertase subtilisin/kexin type 9 (PCSK9) represent an unusual but informative exception in that they are relatively common but have large effects on phenotype. Loss-of-function mutations in PCSK9 occur in ~2.5% of African Americans and are associated with large reductions in coronary heart disease (CHD) risk. The development of agents to inhibit PCSK9 demonstrates the utility of translating genetics into clinical therapeutics. Attempts to identify genes responsible for hypercholesterolaemia have used traditional linkage analysis, which requires samples collected from multiple families with defects in the same gene, or genome-wide association, which requires thousands of samples from the population. More recently, whole-exome sequencing studies have revealed loss-of-function mutations in ANGPTL3 associated with pan-hypolipidemia, and in APOC3 that confer protection against CHD. The application of whole-exome sequencing to large populations or to carefully selected patients can streamline the discovery of causal genetic mutations.


Pitfalls in the Serological Diagnosis of Primary Human Cytomegalovirus Infection in Pregnancy Due to Different Kinetics of IgM Clearance and IgG Avidity Index Maturation.
Primary infection occurs when seronegative women are infected by human cytomegalovirus (HCMV). Diagnosis of primary infection is based on the following: antibody seroconversion, presence of IgM and low IgG avidity index (AI), and presence of DNAemia. The kinetics of HCMV-specific IgM antibody and maturation of AI might be very rapid or long-lasting during primary infection, which makes serological diagnosis insidious. The aims of this study were as follows: (i) to report atypical kinetics of HCMV-specific IgM antibody and AI early after onset of primary HCMV infection in a population of pregnant women, and (ii) to assess the frequency of such results. Altogether, 1309 sequential serum samples collected from 465 pregnant women with primary HCMV infection were included in the study. As a general rule, using the LIAISON®CMVIgMII and LIAISON®CMVIgGAvidityII assays, virus-specific IgM antibody levels decreased, while IgG AI increased over time during the first three months after infection onset. However, early clearance of IgM antibody and/or early IgG AI maturation occurred in 46/426 (10.7%) women. In more details, 20/426 (4.7%) and 26/418 (6.2%) women had undetectable IgM antibody or high IgG AI, respectively, when tested within 1-3 months after well-defined infection onset. Twenty sera from as many women with high IgG AI by the LIAISON assay were further tested for IgG AI by VIDAS®CMVIgGAvidityII and Mikrogen recomLineCMVIgG Avidity assays. Comparable results were obtained with VIDAS, whereas 14/20 sera gave low AI with the Mikrogen assay. In conclusion, about 11% of pregnant women undergoing a primary HCMV infection showed misleading serological results. Additional and appropriate testing might help in reducing the risk of missing HCMV primary infection in pregnancy. Furthermore, preconceptional testing should be strongly recommended.


Evaluation the result of warm normal saline irrigation in ureteral endoscopic surgeries.
Transurethral lithotripsy (TUL) is a major modality for the endoscopic management of ureteral stones. Ureteral spasm makes access for ureters difficult, which causes impaction of the ureteroscope, ureteral dislodge, and a low success rate of endoscopic surgeries. This study described the outcomes of a new endoscopic surgical experience by use of 40-degree warm saline irrigation during TUL compaired with routine ambient air irrigation in TUL. In this randomized clinical trial from 2014 to 2015, 150 patients with ureteral stone with balanced randomization were divided into two parallel groups. Patients underwent TUL in the first group with 20-25 degree saline irrigation and in the second group with 40-degree saline irrigation. One surgical team with the same semi-rigid instrument performed all TULs and the other steps were similar in both groups. Complete stone fragmentation was measured as the primary outcome and the duration of procedure, retrograde stone migration and all and any intraoperative complications were the secondary measurements. While comparing warm saline irrigation with cold saline irrigation, the rate of access to upper ureter was 95% versus 72%, stone retropulsion frequency was 10.7% versus 30.7% and the stone-free rate was 96% versus 76% respectively (P < .05). There was no ureteroscope impaction and ureteral dislodge in both groups. Using warm saline irrigation in endoscopic surgeries results in better surgical outcomes including a lower ureteral spasm rate, greater ureteral muscle relaxation and better access to the upper ureteral zone, and a lower rate of complications, such as ureteroscope impaction, ureteral dislodge and stone retropulsion.


Treatment of atlantoaxial dislocations among patients with cervical osseous or vascular abnormalities utilizing hybrid techniques.
OBJECTIVE Most cervical fixations for atlantoaxial dislocation (AAD) are bilateral and symmetric; however, in the setting of osseous and vascular deformity at the craniovertebral junction, asymmetrical and hybrid fixations are used as "salvage" techniques. Because of the rarity of these cases, hybrid cervical fixations for AAD have not been fully explored. The aim of this study was to evaluate the clinical feasibility and outcomes of posterior hybrid cervical fixations for AAD. METHODS Twenty-one AAD cases were retrospectively studied; 18 had cervical myelopathy with Japanese Orthopaedic Association (JOA) scores ranging from 9 to 16 (mean 13.5). Hybrid fixation techniques included unilateral pedicle screws, transarticular screws, C-2 laminar screws, cervical lateral mass screws, and spinous process screws. During the same period, 82 AAD cases, treated using symmetric traditional fixations, were analyzed as controls. RESULTS Atlantoaxial fixation was performed in 11 cases, while occiput-cervical fixation was used in 10 cases. All cases achieved solid osseous fusion. Anatomical reduction was achieved in 20 cases (95.2%). All 18 cases with myelopathy showed postoperative improvement, with JOA scores ranging from 13 to 17 (mean 15.5). Three cases (14.2%) experienced complications, including delayed wound healing, CSF leakage, and fixation loosening. Hybrid fixation techniques showed significantly greater estimated blood loss when compared with controls (208.1 ± 19.30 ml vs 139.63 ± 8.75 ml, p = 0.001). Operative duration (125.38 ± 6.29 min vs 119.41 ± 3.77 min, p = 0.464), complication rates (14.3% vs 4.9%, p = 0.148), and JOA improvement rates (61% ± 7% vs 49% ± 4%, p = 0.161) showed no significant differences. CONCLUSIONS For ADD with osseous or vascular deformity, posterior cervical reduction and stabilization can be achieved using hybrid techniques, resulting in comparable clinical results to symmetric traditional fixation.


A peripheral component interconnect express-based scalable and highly integrated pulsed spectrometer for solution state dynamic nuclear polarization.
High sensitivity, high data rates, fast pulses, and accurate synchronization all represent challenges for modern nuclear magnetic resonance spectrometers, which make any expansion or adaptation of these devices to new techniques and experiments difficult. Here, we present a Peripheral Component Interconnect Express (PCIe)-based highly integrated distributed digital architecture pulsed spectrometer that is implemented with electron and nucleus double resonances and is scalable specifically for broad dynamic nuclear polarization (DNP) enhancement applications, including DNP-magnetic resonance spectroscopy/imaging (DNP-MRS/MRI). The distributed modularized architecture can implement more transceiver channels flexibly to meet a variety of MRS/MRI instrumentation needs. The proposed PCIe bus with high data rates can significantly improve data transmission efficiency and communication reliability and allow precise control of pulse sequences. An external high speed double data rate memory chip is used to store acquired data and pulse sequence elements, which greatly accelerates the execution of the pulse sequence, reduces the TR (time of repetition) interval, and improves the accuracy of TR in imaging sequences. Using clock phase-shift technology, we can produce digital pulses accurately with high timing resolution of 1 ns and narrow widths of 4 ns to control the microwave pulses required by pulsed DNP and ensure overall system synchronization. The proposed spectrometer is proved to be both feasible and reliable by observation of a maximum signal enhancement factor of approximately -170 for (1)H, and a high quality water image was successfully obtained by DNP-enhanced spin-echo (1)H MRI at 0.35 T.


Chemical and physical variations of cannabis smoke from a variety of cannabis samples in New Zealand.
Studies have compared the chemical properties of tobacco smoke to those of cannabis smoke, with the objective of identifying the chemical attributes responsible for the mutagenicity and carcinogenicity of cannabis smoke. Comparative studies have included small sample sizes and produced conflicting results. The aim of this study was to assess the major chemical and physical variations of cannabis smoke across a range of cannabis samples of different potencies and origins, sourced from the illegal market in New Zealand. Twelve cannabis samples were studied ranging from 1.0% to 13.4% delta-9-tetrahydrocannabinol (Δ9THC) content. A smoking machine was used to smoke "joints" (cannabis cigarettes) and the chemical/physical properties of the smoke assessed. The chemical constituents of the smoke extracts were analysed by gas chromatography/mass spectrometry. A range of different chemical constituents (in addition to Δ9THC) were identified and their concentrations estimated. Terpenoids were identified as the major variable in cannabis smoke, showing a 40-fold range in total terpenoid content. Analysis of the total particulate matter showed that significantly different levels of particulate matter were produced between the different cannabis samples, ranging from 14.6 to 66.3 mg/g of cannabis smoked. The Δ9THC delivery efficiency during smoking was also investigated and produced consistent results showing a mean and median of 12.6% and 10.8%, respectively, of the theoretically available Δ9THC (ranging from 7.2% to 28.0%).


Dispersive locomotion of human neutrophils in response to a steroid-induced factor from monocytes.
A monocyte-derived factor that stimulates the locomotion of human neutrophils on an albumin-coated glass surface has been prepared from the culture supernatant of dexamethasone-treated human monocytes and called STMS (steroid-treated monocyte supernatant). A modified cell tracking program has been developed and the parameters of locomotion determined by the analysis of Gail and Boone for cells moving in a persistent random walk. Cells moving in uniform concentrations of STMS, interleukin-8 (IL-8) and N-formyl-methionyl-leucyl-phenylalanine (fMLP) chosen to give a sub-maximal speed of locomotion show persistent, random and constrained random diffusion, respectively, with augmented diffusion coefficients of 0.8 +/- 0.1, 0.14 +/- 0.02 and 0.12 +/- 0.03 microns 2 per second for STMS, IL-8 and fMLP, respectively. The augmented diffusion coefficient and the underlying persistence are therefore sensitive quantitative assay parameters for STMS activity and the qualitative characteristics of locomotion allow STMS activity to be distinguished from that of all other factors tested. The contribution of lowered adhesion to locomotion was examined in a novel tilt-assay, which demonstrated that cells in the presence of STMS, but not other factors, moved down slope with significantly increased speed while maintaining contact with the substratum. The results were interpreted in terms of the bipolar form of STMS-treated cells, contrasting with multipolar forms in response to other agents. This together with low adhesiveness plus an inherent tendency of a single locomotor focus to continue motion in its original direction has been used to explain the difference between response to STMS and other factors.(ABSTRACT TRUNCATED AT 250 WORDS)


Dynamic Hydrostatic Pressure Regulates Nucleus Pulposus Phenotypic Expression and Metabolism in a Cell Density-Dependent Manner.
Dynamic hydrostatic pressure (HP) loading can modulate nucleus pulposus (NP) cell metabolism, extracellular matrix (ECM) composition, and induce transformation of notochordal NP cells into mature phenotype. However, the effects of varying cell density and dynamic HP magnitude on NP phenotype and metabolism are unknown. This study examined the effects of physiological magnitudes of HP loading applied to bovine NP cells encapsulated within three-dimensional (3D) alginate beads. Study 1: seeding density (1 M/mL versus 4 M/mL) was evaluated in unloaded and loaded (0.1 MPa, 0.1 Hz) conditions. Study 2: loading magnitude (0, 0.1, and 0.6 MPa) applied at 0.1 Hz to 1 M/mL for 7 days was evaluated. Study 1: 4 M/mL cell density had significantly lower adenosine triphosphate (ATP), glycosaminoglycan (GAG) and collagen content, and increased lactate dehydrogenase (LDH). HP loading significantly increased ATP levels, and expression of aggrecan, collagen I, keratin-19, and N-cadherin in HP loaded versus unloaded groups. Study 2: aggrecan expression increased in a dose dependent manner with HP magnitude, whereas N-cadherin and keratin-19 expression were greatest in low HP loading compared to unloaded. Overall, the findings of the current study indicate that cell seeding density within a 3D construct is a critical variable influencing the mechanobiological response of NP cells to HP loading. NP mechanobiology and phenotypic expression was also found to be dependent on the magnitude of HP loading. These findings suggest that HP loading and culture conditions of NP cells may require complex optimization for engineering an NP replacement tissue.


Effects of sea ice and wind speed on phytoplankton spring bloom in central and southern Baltic Sea.
In this study, the effects of sea ice and wind speed on the timing and composition of phytoplankton spring bloom in the central and southern Baltic Sea are investigated by a hydrodynamic-biogeochemical model and observational data. The modelling experiment compared the results of a reference run in the presence of sea ice with those of a run in the absence of sea ice, which confirmed that ecological conditions differed significantly for both the scenarios. It has been found that diatoms dominate the phytoplankton biomass in the absence of sea ice, whereas dinoflagellates dominate the biomass in the presence of thin sea ice. The study concludes that under moderate ice conditions (representing the last few decades), dinoflagellates dominate the spring bloom phytoplankton biomass in the Baltic Sea, whereas diatoms will be dominant in the future as a result of climate change i.e. in the absence of sea ice.


Inhibition of PI3K induces Rac activation and membrane ruffling in proto-Dbl expressing cells.
Proto-Dbl protein, a guanine nucleotide exchange factor (GEF) for Rho GTPases, is tightly regulated by a combination of mechanisms that involve intra- and intermolecular interaction and N- and C-terminal domain-dependent turnover of the protein. Moreover, the interaction of the PH domain of proto-Dbl with phosphoinositides regulates its subcellular localization and biological activity. Here we show that inhibition of the phosphatidylinositol 3-kinase (PI3K) by molecular and pharmacological inhibitors causes a strong inhibition of proto-Dbl-induced cell proliferation and transformation. Conversely, inhibition of PI3K results in the translocation of proto-Dbl to the plasma membrane, Rac activation and increased membrane ruffles and cell motility. Furthermore, we investigated the signaling molecules involved in proto-Dbl-induced cell transformation and motility and observed that inhibition of PI3K in proto-Dbl expressing cells induces an increase in p38 activity and a decrease in ERK phosphorylation. Our results suggest that proto-Dbl activates distinct downstream effectors to induce morphological changes and cell transformation.


Physiological and Mechanical Indices Serving the New Cross-Country Olympic Mountain Bike Performance.
To identify relevant physiological, mechanical, and strength indices to improve the evaluation of elite mountain bike riders competing in the current Cross-Country Olympic (XCO) format. Considering the evolution of the XCO race format over the last decade, the present testing protocol adopted a battery of complementary laboratory cycling tests: a maximal aerobic consumption, a force-velocity test, and a multi-short-sprint test. A group of 33 elite-level XCO riders completed the entire testing protocol and at least 5 international competitions. Very large correlations were found between the XCO performance and maximal aerobic power output (r = .78; P < .05), power at the second ventilation threshold (r = .83; P < .05), maximal pedaling force (r = .77; P < .05), and maximum power in the sixth sprint (r = .87; P < .05) of the multi-short-sprint test. A multiple regression model revealed that the normalized XCO performance was predicted at 89.2% (F3,29 = 89.507; r = .95; P < .001) by maximum power in the sixth sprint (β = 0.602; P < .001), maximal pedaling rate (β = 0.309; P < .001), and relative maximal aerobic power output (β = 0.329; P < .001). Confirming our expectations, the current XCO performance was highly correlated with a series of physiological and mechanical parameters reflecting the high level of acyclic and intermittent solicitation of both aerobic and anaerobic metabolic pathways and the required qualities of maximal force and velocity. The combination of physiological, mechanical, and strength characteristics may thus improve the prediction of elite XCO cyclists' performance. It seems of interest to evaluate the ability to repeatedly produce brief intensive efforts with short active recovery periods.


Fostering interprofessional collaborative practice in acute care through an academic-practice partnership.
An interprofessional collaborative practice (IPCP) environment was implemented in four inpatient acute care unit accountable care teams (ACT) through an academic-practice partnership. An action research methodology was implemented to empower healthcare professionals and promote ownership and sustainment of the IPCP strategies. Healthcare professionals on the ACT units, students, and faculty engaged in the multi-year project. Interventions included staff engagement exercises and coaching and support of individual ACT unit IPCP strategies and education. Healthcare professional outcome data were collected in the form of participant surveys, measuring collaboration about care decisions and the extent to which professionals engaged in a culture of safety, collected at baseline and 6-month intervals. Healthcare outcome measures of the ACT units were also collected at baseline and 6-month intervals. Students had clinical learning experiences in the IPCP settings and completed post clinical surveys. Implementation of the interventions resulted in a positive trend in the healthcare professionals, although statistical significance was not observed. Student outcome data demonstrated statistically significant positive learning outcomes. Healthcare outcome measures demonstrated a significant decrease in readmissions and an overall decrease in catheter-associated urinary tract infections over time. Other measures were not significantly impacted. In conclusion, an academic - practice partnership can strengthen and support an IPCP environment by allowing healthcare clinicians to be directly involved in the selection and implementation of IPCP strategies and contribute to improved professional, healthcare and student outcomes.


Rapid and marker-free gene replacement in citric acid-producing Aspergillus tubingensis (A. niger) WU-2223L by the CRISPR/Cas9 system-based genome editing technique using DNA fragments encoding sgRNAs.
Strains belonging to Aspergillus section Nigri, including Aspergillus niger, are used for industrial production of citric acid from carbohydrates such as molasses and starch. The objective of this study was to construct the genome editing system that could enable rapid and efficient gene replacement in citric acid-producing fungi for genetic breeding. Using the citric acid-hyperproducer A. tubingensis (formerly A. niger) WU-2223L as a model strain, we developed a CRISPR/Cas9 system-based genome editing technique involving co-transformation of Cas9 and the DNA fragment encoding single guide RNA (sgRNA). Using this system, ATP-sulfurylase gene (sC) knock-out strain derived from WU-2223L was generated; the knock-out efficiency was 29 transformants when 5 μg Cas9 was added to 5 × 105 protoplasts. In the gene replacement method based on this system, a DNA fragment encoding sgRNAs that target both the gene of interest and marker gene was used, and replacement of nitrate reductase gene (niaD) using sC gene as a marker gene was attempted. More than 90% of the sC-knock-out transformants exhibited replaced niaD, indicating efficient gene replacement. Moreover, one-step marker rescue of the sC marker gene was accomplished by excising the knock-in donor via intramolecular homologous recombination, enabling marker-free genome editing and drastically shortening the gene replacement period by circumventing the transformation procedure to recover the sC gene. Thus, we succeeded in constructing a CRISPR/Cas9 system-based rapid and marker-free gene replacement system for the citric acid-hyperproducer strain WU-2223L.


Food insecurity, morbidity, and susto: Factors associated with depression severity in Guatemala measured with the Personal Health Questionnaire 9.
Depression is the largest contributor to non-fatal health loss globally and the majority of this burden occurs in low- and middle-income countries. Yet, estimates of prevalence rates and severity in these contexts may be uncertain due to limited screening, lack of mental health providers, and stigma around mental disorders which may prevent individuals from seeking care. In Guatemala, estimates of depression vary, due in part to the range of screening and diagnostic instruments used and diversity of sample populations. Most studies emphasize personal experiences with violence as a predictor of depression in Guatemala, although high rates of inequality, discrimination, and resource scarcity in the country potentially play a role. In this study, we examine factors associated with depression severity categories measured with the Personal Health Questionnaire 9 (PHQ-9) among a random sample of women in a small urban community in the Central Highlands of Guatemala. Participants were recruited through a randomized sample of households in a small urban community. Participants completed a questionnaire which included questions on demographics, illness history, food insecurity, and the PHQ-9. In total, 101 women were included in the analysis. Food insecurity, 2-week symptom reporting, and experiencing susto are associated with higher depression severity categories. This research highlights need for more research on factors related to the prevalence and severity of mental disorders, and the relationship between mental disorders and cultural constructs of distress, particularly in areas like Guatemala with limited mental health services.


Measurement of Penrose Superradiance in a Photon Superfluid.
The superradiant amplification in the scattering from a rotating medium was first elucidated by Sir Roger Penrose over 50 years ago as a means by which particles could gain energy from rotating black holes. Despite this fundamental process being ubiquitous also in wave physics, it has only been observed once experimentally, in a water tank. Here, we measure this amplification for a nonlinear optics experiment in the superfluid regime. In particular, by focusing a weak optical beam carrying orbital angular momentum onto the core of a strong pump vortex beam, negative norm modes are generated and trapped inside the vortex core, allowing for amplification of a reflected beam. Our experiment demonstrates amplified reflection due to a novel form of nonlinear optical four-wave mixing, whose phase-relation coincides with the Zel'dovich-Misner condition for Penrose superradiance in our photon superfluid, and unveil the role played by negative frequency modes in the process.


Predicting carcass and body fat composition using biometric measurements of grazing beef cattle.
This study was conducted to develop equations to predict carcass and body fat compositions using biometric measures (BM) and body postmortem measurements and to determine the relationships between BM and carcass fat and empty body fat compositions of 44 crossbred bulls under tropical grazing conditions. The bulls were serially slaughtered in 4 groups at approximately 0 d (n = 4), 84 d (n = 4), 168 d (n = 8), 235 d (n = 8), and 310 d (n = 20) of growth. The day before each slaughter, bulls were weighed, and BM were taken, including hook bone width, pin bone width, abdomen width, body length, rump height, height at withers, pelvic girdle length, rib depth, girth circumference, rump depth, body diagonal length, and thorax width. Others measurements included were total body surface (TBS), body volume (BV), subcutaneous fat (SF), internal fat (InF), intermuscular fat, carcass physical fat (CFp), empty body physical fat (EBFp), carcass chemical fat (CFch), empty body chemical fat (EBFch), fat thickness in the 12th rib (FT), and 9th- to 11th-rib section fat (HHF). The stepwise procedure was used to select the variables included in the model. The r(2) and the root-mean-square error (RMSE) were used to account for precision and variability. Our results indicated that lower rates of fat deposition can be attributed to young cattle and low concentration of dietary energy under grazing conditions. The BM improved estimates of TBS (r(2) = 0.999) and BV (r(2) = 0.997). The adequacy evaluation of the models developed to predict TBS and BV using theoretical equations indicated precision, but lower and intermediate accuracy (bias correction = 0.138 and 0.79), respectively, were observed. The data indicated that BM in association with shrunk BW (SBW) were precise in accounting for variability of SF (r(2) = 0.967 and RMSE = 0.94 kg), InF (r(2) = 0.984 and RMSE = 1.26 kg), CFp (r(2) = 0.981 and RMSE = 2.98 kg), EBFp (r(2) = 0.985 and RMSE = 3.99 kg), CFch (r(2) = 0.940 and RMSE = 2.34 kg), and EBFch (r(2) = 0.934 and RMSE = 3.91 kg). Results also suggested that approximately 70% of body fat was deposited as CFp and 30% as InF. Furthermore, the development of an equation using HHF as a predictor, in combination with SBW, was a better predictor of CFp and EBFp than using HHF by itself. We concluded that the prediction of physical and chemical CF and EBF composition of grazing cattle can be improved using BM as a predictor.


Treatment changes in a cohort of people with apparently drug-resistant epilepsy: an extended follow-up.
The seizure response to the addition of a previously unused antiepileptic drug in a cohort of 155 people with refractory epilepsy was previously reported after a median of 18 months follow-up. The authors followed 139 (90%) of the original cohort for a median follow-up of 6.9 years to determine the longer term outcome in people with refractory epilepsy. During the 6.9 year follow-up period, a total of 448 medication changes were made. Eight per cent of these resulted in 12 months or more of seizure freedom and a further 17% of changes resulted in at least 50% improvement in seizure frequency. At the last follow-up, 26 (19%) of individuals had been seizure-free for 12 months or more, and 41 (29%) had 50%-99% improvement in seizure frequency. Terminal seizure freedom was correlated with having no seizures at the time of the previous report (p=0.03), a lower number of previous antiepileptic drugs taken (p=0.052) and a lower number of concomitant antiepileptic drugs (p=0.03). In those who entered remission the probability of remaining seizure-free 5 years later was 0.48 (95% CI 0.32 to 0.63). This suggests that about half of people with apparent drug-resistant epilepsy can have significant improvements in seizure control with further drug changes. Some will subsequently relapse, but long periods of seizure freedom or significantly improved seizure control in the absence of complete seizure control can occur. Such valuable improvements suggest that the recently proposed International League against Epilepsy definition of refractory epilepsy may be too restrictive.


Poly-L-lactic acid as a facial filler.
Poly-L-lactic acid is a filler recently approved by the US FDA for the correction of facial lipoatrophy in patients infected with the human immunodeficiency virus (HIV). Currently, poly-L-lactic acid, sold under the brand name Sculptratrade mark (Dermik), is the only product approved by the FDA specifically for this indication. The market for poly-L-lactic acid will likely be larger than the HIV-infected population, as physicians use poly-L-lactic acid off-label to correct lipoatrophy associated with the normal aging process in non-HIV-infected patients. The benefits of poly-L-lactic acid are limited by the fact that multiple treatments are necessary to achieve the desired correction; its results are temporary and its cost is high.


Rectal Cancer Survivors' Participation in Productive Activities.
Rectal cancer and its treatment impair survivors' productivity. To assess determinants of market and nonmarket employment, job search, volunteering, and homemaking among survivors five years or longer after diagnosis. We mailed questionnaires to 1063 survivors who were members of Kaiser Permanente (Northern California, Northwest) during 2010 and 2011. Productive activities, functional health status, and bowel management at the time of the survey. Response rate was 60.5% (577/953). Higher comorbidity burdens were associated with lower productivity for men and women rectal cancer survivors. Productive survivors were younger and had lower disease stage and age at diagnosis, higher household income and educational attainment, and fewer comorbidity burdens and workplace adjustments than did nonproductive survivors (p < 0.05 each; 2-sided). Productive rectal cancer survivors were evenly split by sex. Staying productive is associated with better mental health for rectal cancer survivors. Rectal cancer survivors with multiple chronic conditions, higher disease stage, lower productive activities, and older age need better access to medical care and closer monitoring of the quality of their care, including self-care. To capture the full extent of the involvement of survivors in all types of productive activities, research should routinely include measures of employment, searching for employment, homemaking, and volunteering. Counting market and nonmarket productive activities is innovative and recognizes the continuum of contributions survivors make to families and society. Health care systems should routinely monitor rectal cancer survivors' medical care access, comorbidities, health-related quality of life, and productive activities.


Polysorbate 20 prevents the precipitation of a monoclonal antibody during shear.
Monoclonal antibodies (MAbs) are widely used as therapeutic proteins and they are frequently exposed to a high degree of stress during manufacturing or delivery. MAbs shear thin upon increasing shear rates. After undergoing multiple shear cycles, with a cone-and-plate rheometer, the solution viscosity of high concentration antibodies increases due to the formation of insoluble aggregates. These shear-induced insoluble aggregates do not form when polysorbate 20 is present in solution. We hypothesize that monoclonal antibodies form a thin protein layer at the air-water interface. MAbs at the interface expose their hydrophobic core to air leading to unfolding, multiple non-specific intermolecular interactions and, upon continuous high shear, precipitation. Surface tension analysis confirms that monoclonal antibodies are surface active and that polysorbate 20 can prevent their interaction with the air-water interface. In addition, we complement these findings with a viscometer that measures bulk viscosity without the influence of an air-liquid interfacial viscosity and find that the bulk viscosity increases slightly when Mab solutions contained polysorbate 20. These methods of analysis could be used when designing manufacturing systems in which a protein solution is subject to shear forces.


Two-body Schrödinger wave functions in a plane-wave basis via separation of dimensions.
Using a combination of ideas, the ground and several excited electronic states of the helium atom and the hydrogen molecule are computed to chemical accuracy-i.e., to within 1-2 mhartree or better. The basic strategy is very different from the standard electronic structure approach in that the full two-electron six-dimensional (6D) problem is tackled directly, rather than starting from a single-electron Hartree-Fock approximation. Electron correlation is thus treated exactly, even though computational requirements remain modest. The method also allows for exact wave functions to be computed, as well as energy levels. From the full-dimensional 6D wave functions computed here, radial distribution functions and radial correlation functions are extracted-as well as a 2D probability density function exhibiting antisymmetry for a single Cartesian component. These calculations support a more recent interpretation of Hund's rule, which states that the lower energy of the higher spin-multiplicity states is actually due to reduced screening, rather than reduced electron-electron repulsion. Prospects for larger systems and/or electron dynamics applications appear promising.


Pre-weaning management of calves on commercial dairy farms and its influence on calf welfare and mortality.
Welfare and management of calves is of increasing interest and also influences performance of these animals in later life. The aim of this study was to assess management and environmental conditions under which pre-weaned dairy calves are reared on commercial Irish dairy farms. We included 47 spring-calving, pasture-based herds in this study. Herd and animal-specific data, such as mortality rate, age and breed, were gathered from all participants via the HerdPlus® database. Information pertaining to management practices was collected by conducting an interview with the principal calf rearer, while an assessment of calf housing facilities was conducted to identify conditions calves were reared in. The environmental assessment included measurements of space allowance per calf, as well as feeding equipment hygiene. To assess calf behaviour video observations were used, while accounting for the number of calves present in a group and the space available per calf. Faecal samples were also collected to determine the presence of enteric pathogens among calves. To compare calf space allowance, group size and presence of enteric pathogens early and late in the calving season each farm was visited twice. Calf mortality was not associated with either herd size, space allowance per calf or post-colostrum feeding practices. Higher calf mortality was identified among herds which reported experiencing an on-set of calf pneumonia during weeks 8 to 10 of the calving season. This study demonstrates that factors associated with calf welfare on commercial Irish dairy farms (e.g. space allowance, mortality rate) are independent of herd size. Some management practices however, such as methods used for treating health issues can affect rates of calf mortality experienced. Calf mortality, for example, was lower in herds which treated diarrhoea cases by administering electrolytes, while continuing to offer milk. Behavioural observations indicate that smaller group sizes could promote expression of positive behaviours, potentially resulting from an overall improvement in welfare. Space allowance per calf was not associated with observed behaviour frequencies. We also identified that similar rates of calf mortality are experienced across herds of different sizes.


Investigating key cell types and molecules dynamics in PyMT mice model of breast cancer through a mathematical model.
The most common kind of cancer among women is breast cancer. Understanding the tumor microenvironment and the interactions between individual cells and cytokines assists us in arriving at more effective treatments. Here, we develop a data-driven mathematical model to investigate the dynamics of key cell types and cytokines involved in breast cancer development. We use time-course gene expression profiles of a mouse model to estimate the relative abundance of cells and cytokines. We then employ a least-squares optimization method to evaluate the model's parameters based on the mice data. The resulting dynamics of the cells and cytokines obtained from the optimal set of parameters exhibit a decent agreement between the data and predictions. We perform a sensitivity analysis to identify the crucial parameters of the model and then perform a local bifurcation on them. The results reveal a strong connection between adipocytes, IL6, and the cancer population, suggesting them as potential targets for therapies.


Disease Management Evaluation: A Comprehensive Review of Current State of the Art.
Many countries across Europe and elsewhere have been experimenting with various structured approaches to manage patients with chronic illness as a way to improve quality of care, reduce costs and lead to better population health outcomes in the long run. Despite a body of studies of disease management interventions, uncertainty about the effects of these remains not least because current guidance on evaluation methods and metrics require further development to enhance scientific rigour while also being practical in routine operations. This article provides details from a report that reviews the academic and grey literature to help advance the task of improving the science of assessing disease management initiatives in Europe. Challenges identified are methodological, analytical and conceptual in nature, with a key issue being the establishment of the counterfactual. An array of sophisticated statistical techniques and analytical frameworks can assist in the construction of a sound comparison strategy when a randomised controlled trial is not possible. Issues to consider include: a clear framework of the mechanisms of action and expected effects of disease management; an understanding of the characteristics of disease management (scope, content, dose, context), and of the intervention and target populations (disease type, severity, case-mix); a period of observation over multiple years; and a logical link between performance measures and the intervention's aims and underlying theory of behaviour change.


Proteomic associations with forced expiratory volume - a Mendelian randomisation study.
A decline in forced expiratory volume (FEV1) is a hallmark of obstructive respiratory diseases, an important cause of morbidity among the elderly. While some data exist on biomarkers that are related to FEV1, we sought to do a systematic analysis of causal relations of biomarkers with FEV1. Data from the general population-based AGES-Reykjavik study were used. Proteomic measurements were done using 4,782 DNA aptamers (SOMAmers). Data from 1,648 participants with spirometric data were used to assess the association of SOMAmer measurements with FEV1 using linear regression. Bi-directional Mendelian randomisation (MR) analyses were done to assess causal relations of observationally associated SOMAmers with FEV1, using genotype and SOMAmer data from 5,368 AGES-Reykjavik participants and genetic associations with FEV1 from a publicly available GWAS (n = 400,102). In observational analyses, 473 SOMAmers were associated with FEV1 after multiple testing adjustment. The most significant were R-Spondin 4, Alkaline Phosphatase, Placental Like 2 and Retinoic Acid Receptor Responder 2. Of the 235 SOMAmers with genetic data, eight were associated with FEV1 in MR analyses. Three were directionally consistent with the observational estimate, Thrombospondin 2 (THBS2), Endoplasmic Reticulum Oxidoreductase 1 Beta and Apolipoprotein M. THBS2 was further supported by a colocalization analysis. Analyses in the reverse direction, testing whether changes in SOMAmer levels were caused by changes in FEV1, were performed but no significant associations were found after multiple testing adjustments. In summary, this large scale proteogenomic analyses of FEV1 reveals protein markers of FEV1, as well as several proteins with potential causality to lung function.


Plasma Membrane Lipids: An Important Binding Site for All Lipoprotein Classes.
Cholesterol is one of the main constituents of plasma membranes; thus, its supply is of utmost importance. This review covers the known mechanisms of cholesterol transfer from circulating lipoprotein particles to the plasma membrane, and vice versa. To achieve homeostasis, the human body utilizes cellular de novo synthesis and extracellular transport particles for supply of cholesterol and other lipids via the blood stream. These lipoprotein particles can be classified according to their density: chylomicrons, very low, low, and high-density lipoprotein (VLDL, LDL, and HDL, respectively). They deliver and receive their lipid loads, most importantly cholesterol, to and from cells by several redundant routes. Defects in one of these pathways (e.g., due to mutations in receptors) usually are not immediately fatal. Several redundant pathways, at least temporarily, compensate for the loss of one or more of them, but the defects trigger systemic diseases, such as atherosclerosis later on. Recently, intracellular membrane-membrane contact sites were shown to be involved in intracellular cholesterol transfer and the plasma membrane itself has been proposed to act as a binding site for lipoprotein-mediated cargo unloading.


Heterotypic binding between neuronal membrane vesicles and glial cells is mediated by a specific cell adhesion molecule.
By means of a multistage quantitative assay, we have identified a new kind of cell adhesion molecule (CAM) on neuronal cells of the chick embryo that is involved in their adhesion to glial cells. The assay used to identify the binding component (which we name neuron-glia CAM or Ng-CAM) was designed to distinguish between homotypic binding (e.g., neuron to neuron) and heterotypic binding (e.g., neuron to glia). This distinction was essential because a single neuron might simultaneously carry different CAMs separately mediating each of these interactions. The adhesion of neuronal cells to glial cells in vitro was previously found to be inhibited by Fab' fragments prepared from antisera against neuronal membranes but not by Fab' fragments against N-CAM, the neural cell adhesion molecule. This suggested that neuron-glia adhesion is mediated by specific cell surface molecules different from previously isolated CAMs . To verify that this was the case, neuronal membrane vesicles were labeled internally with 6-carboxyfluorescein and externally with 125I-labeled antibodies to N-CAM to block their homotypic binding. Labeled vesicles bound to glial cells but not to fibroblasts during a 30-min incubation period. The specific binding of the neuronal vesicles to glial cells was measured by fluorescence microscopy and gamma spectroscopy of the 125I label. Binding increased with increasing concentrations of both glial cells and neuronal vesicles. Fab' fragments prepared from anti-neuronal membrane sera that inhibited binding between neurons and glial cells were also found to inhibit neuronal vesicle binding to glial cells. The inhibitory activity of the Fab' fragments was depleted by preincubation with neuronal cells but not with glial cells. Trypsin treatment of neuronal membrane vesicles released material that neutralized Fab' fragment inhibition; after chromatography, neutralizing activity was enriched 50-fold. This fraction was injected into mice to produce monoclonal antibodies; an antibody was obtained that interacted with neurons, inhibited binding of neuronal membrane vesicles to glial cells, and recognized an Mr = 135,000 band in immunoblots of embryonic chick brain membranes. These results suggest that this molecule is present on the surfaces of neurons and that it directly or indirectly mediates adhesion between neurons and glial cells. Because the monoclonal antibody as well as the original polyspecific antibodies that were active in the assay did not bind to glial cells, we infer that neuron-glial interaction is heterophilic, i.e., it occurs between Ng-CAM on neurons and an as yet unidentified CAM present on glial cells.


LPS responsiveness and neutrophil chemotaxis in vivo require PMN MMP-8 activity.
We identify matrix metalloproteinase (MMP)-8, the polymorphonuclear (PMN) leukocyte collagenase, as a critical mediator initiating lipopolysaccharide (LPS)-responsiveness in vivo. PMN infiltration towards LPS is abrogated in Mmp8-null mice. MMP-8 cleaves LPS-induced CXC chemokine (LIX) at Ser(4)-Val(5) and Lys(79)-Arg(80). LIX bioactivity is increased upon N-terminal cleavage, enhancing intracellular calcium mobilization and chemotaxis upon binding its cognate receptor, CXCR2. As there is no difference in PMN chemotaxis in Mmp8-null mice compared with wild-type mice towards synthetic analogues of MMP-8-cleaved LIX, MMP-8 is not essential for extravasation or cell migration in collagenous matrices in vivo. However, with biochemical redundancy between MMPs 1, 2, 9, and 13, which also cleave LIX at position 4 approximately 5, it was surprising to observe such a markedly reduced PMN infiltration towards LPS and LIX in Mmp8-/- mice. This lack of physiological redundancy in vivo identifies MMP-8 as a key mediator in the regulation of innate immunity. Comparable results were found with CXCL8/IL-8 and CXCL5/ENA-78, the human orthologues of LIX. MMP-8 cleaves CXCL8 at Arg(5)-Ser(6) and at Val(7)-Leu(8) in CXCL5 to activate respective chemokines. Hence, rather than collagen, these PMN chemoattractants are important MMP-8 substrates in vivo; PMN-derived MMP-8 cleaves and activates LIX to execute an in cis PMN-controlled feed-forward mechanism to orchestrate the initial inflammatory response and promote LPS responsiveness in tissue.


Reduction in Endometrioma Size with Three Months of Aromatase Inhibition and Progestin Add-Back.
The purpose of this study was to assess the impact of 3 months of aromatase inhibition together with progestin add-back on ovarian endometrioma size. This prospective cohort study was performed at University Medical Center (UC San Diego). Women trying to conceive were excluded. After informed consent, all women were treated with the aromatase inhibitor letrozole (5 mg/d) with norethindrone acetate (5 mg/d) add-back for 3 months. Pre- and posttreatment assessments of endometrioma sizes were performed by ultrasound. The impact of treatment on pain was determined using the patient assessed endpoints of the Biberoglu and Behrman scale. These included assessing dysmenorrhea, dyspareunia, and nonmenstrual pelvic pain each on a scale from 0 to 3. The primary endpoint of this study was the change in ultrasound documented endometrioma size. Fourteen endometriomas in 8 consecutive women were treated for 3 m. Mean endometrioma diameter decreased 50% from 4.6 ± 1.6 cm to 2.3 ± 1.6 cm (mean ± SD). This represents a 75% decrease in endometrioma volume. Endometriosis symptoms of dysmenorrhea, dyspareunia, and nonmenstrual pelvic pain also improved with treatment. In conclusion, a 3-month course of high dose aromatase inhibition with progestin add-back significantly reduces ovarian endometrioma size and warrants further investigation.


Detection and Identification of Reaction Intermediates in the Photorearrangement of Pyridazine N-Oxide: Discrepancies between Experiment and Theory.
Photolysis of pyridazine N-oxide (PNO) results in the detection of a complex series of transient phenomena. On the ultrafast (fs) timescale, we could detect the decay of the first singlet excited state of PNO and the formation of a short-lived transient species, which, based on its time-resolved resonance Raman (TR3) spectrum, we assign to oxaziridine 1,2-diaza-7-oxa-bicyclo[4.1.0]hepta-2,4-diene. On a longer (hundreds of ns) timescale, this species rearranges to a ring-opened diazo compound, which we have also detected by time-resolved infrared and TR3 spectroscopy. In addition, we identify 1-oxa-3,4-diazepine as a long-lived species formed in the photochemistry of PNO. This species is formed via its oxirane isomer, which in turn is likely formed directly from the S1 state of PNO via a conical intersection. The barrier determined experimentally for the decay of 1,2-diaza-7-oxa-bicyclo[4.1.0]hepta-2,4-diene (Ea = (7.1 ± 0.5) kcal mol-1) is far larger than any barrier calculated by any method that includes dynamic electron correlation but very close to the barriers calculated at the RHF or CASSCF levels of theory. Many methods (B3LYP, MP2, and MP4) fail to give a minimum structure for 1,2-diaza-7-oxa-bicyclo[4.1.0]hepta-2,4-diene, while M06, M06-2X, MP3, CCSD, or CCSD(T) yield activation energies for its electrocyclic ring opening that are far too small. In addition, we note that several important geometric parameters, both of 1,2-diaza-7-oxa-bicyclo[4.1.0]hepta-2,4-diene and of the transition state of its ring opening reaction, clearly have reached no convergence, even at the fully optimized CCSD(T)/cc-pVTZ level of theory. We therefore suggest that the transient species described in this contribution might be excellent test molecules for further development of highly correlated and density functional theory methods.


Singular or plural? Children's knowledge of the factors that determine the appropriate form of count nouns.
Two experiments investigated the factors that govern children's use of singular and plural forms of count nouns. Experiment 1 used an elicited production task to investigate whether children use referential and/or syntactic information to determine the form of the count nouns when the two sources of information conflict (e.g. each x, one of the xs), as well as when the linguistic context does not provide any constraint on the form of the noun, but the referential context does (e.g. the dog(s)). 48 children, aged 1;9 to 5;6, participated in Experiment 1. The results suggest that even the youngest children can use referential information when relevant, and can ignore referential information when necessary. Children did, however, show a tendency to make errors with the quantifier each in non-partitive contexts, and a developmental trend was found in the use and avoidance of each in non-partitive contexts. Experiment 2, an act out task, provided a second test of the role of referential information in children's use of singular and plural forms. Experiment 2 also investigated children's appreciation of the semantic distinction between each and all. 48 children, aged 1;8 to 5;6, participated in Experiment 2. A weak sensitivity to the semantic distinction between the two quantifiers as well as the syntactic context in which they were used was found. The results of the two experiments suggest that, from the beginning, children approach the task of learning when to use singular and plural forms of count nouns on the basis of morphosyntactic, semantic, and referential properties of utterances, rather than initially using only one of these types of information.


Associations Between 30-Day Mortality, Specialist Nursing, and Daily Physician Ward Rounds in a National Stroke Registry.
Background and Purpose- Well-organized stroke care is associated with better patient outcomes, but the most important organizational factors are unknown. Methods- Data were extracted from the Sentinel Stroke National Audit Programme of adults with acute stroke treated in stroke hospitals in England and Wales between April 2013 and March 2015. Multilevel models with random intercepts for hospitals were used to estimate the association of each variable with 30-day mortality to estimate the impact of admission to differently organized hospitals. Results- Of the 143 578 patients with acute stroke admitted to 154 hospitals, 14.4% died within 30 days of admission. In adjusted analyses, admission to hospitals with higher ratios of nurses trained in swallow screening was associated with reduced odds of death ( P=0.004), and admission to hospitals with daily physician ward rounds was associated with 10% lower odds of mortality compared with less-frequent ward rounds (95% CI, 0.82-0.98; P=0.013). Number of stroke admissions and overall ratio of registered nurses on duty at weekends were not found to be independently associated with mortality after adjustment for other factors. Conclusions- If these associations are causal, an extra 1332 deaths annually in England and Wales could be saved by hospitals providing care associated with a ratio of nurses trained in swallow screening of at least 3 per 10 beds and daily stroke physician ward rounds.


New Wide Band Gap Donor for Efficient Fullerene-Free All-Small-Molecule Organic Solar Cells.
A new organic small molecule, DRTB-T, that incorporates a two-dimensional trialkylthienyl-substituted benzodithiophene core building block was designed and synthesized. DRTB-T has a band gap (Egopt) of 2.0 eV with a low-lying highest occupied molecular orbital (HOMO) level of -5.51 eV. Nonfullerene small-molecule solar cells consisting of DRTB-T and a nonfullerene acceptor (IC-C6IDT-IC) were constructed, and the morphology of the active layer was fine-tuned by solvent vapor annealing (SVA). The device showed a record 9.08% power conversion efficiency (PCE) with a high open-circuit voltage (Voc = 0.98 V). This is the highest PCE for a nonfullerene small-molecule organic solar cell (NFSM-OSC) reported to date. Our notable results demonstrate that the molecular design of a wide band gap (WBG) donor to create a well-matched donor-acceptor pair with a low band gap (LBG) nonfullerene small-molecule acceptor, as well as subtle morphological control, provides great potential to realize high-performance NFSM-OSCs.


Do cervical stabilization exercises change the effects of conventional exercises in patients with Parkinson's disease?
The aim of this study was to examine whether cervical stabilization exercises (CSEs) change the effects of conventional exercises (CEs) in patients with PD. Twenty-five patients with PD were randomized into two groups. While the experimental group (EG) received CSEs in addition to CEs, the control group (CG) received only CEs. Both programs lasted 8 weeks. Eighteen participants were able to complete the study. The outcomes were the changes in posture, cervical joint position sense (JPS), balance assessment, 10-m walking tests (10MWT), and the Timed Up & Go (TUG) test. At the end of the study, significant improvement was observed in both groups in terms of trunk rotation angle and pelvic asymmetry, the time parameter of TUG, and the 10MWT (p < 0.05). In the EG, greater improvement was detected in the Berg Balance Scale, static posturography, postural alignment, JPS, and the cadence parameter of TUG (p < 0.05). CEs and CSEs could improve walking speed and posture in patients with PD, but if CSEs are added to CEs, greater improvements could be achieved in JPS and postural control.(Clinical Trials ID: NCT03854747).


Incidence and pathological features of spontaneous tumors in athymic nude mice.
Systematic observation of 1141 nude mice (Swiss background [corrected] strain) that received human tumor transplants revealed 24 spontaneous tumors, 18 of lymphoreticular origin and 6 pulmonary adenomas. Spontaneous tumors were seen at an average age of 9.1 months, and 22 of the tumors were seen only in that fraction of our group (324 mice) surviving for 5 months or more (22 of 324 X 100 = 6.8%). Transplantation of these tumors to other nude mice was successful in three of five cases. Mice transplanted with adenocarcinoma of the colon and with tumors of the urogenital tract developed spontaneous tumors more often than did mice receiving other types of human tumor transplants. Progressive growth of the human tumor transplant occurred significantly less often in the mice that eventually developed spontaneous tumors than in the mice that showed no spontaneous tumor development. Nevertheless, the incidence of spontaneous tumors in these nude mice was similar to that reported for the thymus-bearing background strain.


Inhibition of mutant RAS-RAF interaction by mimicking structural and dynamic properties of phosphorylated RAS.
Undruggability of RAS proteins has necessitated alternative strategies for the development of effective inhibitors. In this respect, phosphorylation has recently come into prominence as this reversible post-translational modification attenuates sensitivity of RAS towards RAF. As such, in this study, we set out to unveil the impact of phosphorylation on dynamics of HRASWT and aim to invoke similar behavior in HRASG12D mutant by means of small therapeutic molecules. To this end, we performed molecular dynamics (MD) simulations using phosphorylated HRAS and showed that phosphorylation of Y32 distorted Switch I, hence the RAS/RAF interface. Consequently, we targeted Switch I in HRASG12D by means of approved therapeutic molecules and showed that the ligands enabled detachment of Switch I from the nucleotide-binding pocket. Moreover, we demonstrated that displacement of Switch I from the nucleotide-binding pocket was energetically more favorable in the presence of the ligand. Importantly, we verified computational findings in vitro where HRASG12D/RAF interaction was prevented by the ligand in HEK293T cells that expressed HRASG12D mutant protein. Therefore, these findings suggest that targeting Switch I, hence making Y32 accessible might open up new avenues in future drug discovery strategies that target mutant RAS proteins.


Enhancement of organic solar cell performance by incorporating gold quantum dots (AuQDs) on a plasmonic grating.
The incorporation of metallic nanoobjects into devices allows to increase light harvesting, which increases the device performance. In this study, we used a combination of gold quantum dots and grating-coupled surface plasmon resonance (GCSPR) to improve the performance of organic solar cells (OSCs) with a poly(3-hexylthiophene-2,5-diyl) (P3HT):[6,6]-phenyl C61 butyric acid methyl ester (PCBM) photoactive layer. Gold quantum dots with a green fluorescent color (green-AuQD) were loaded into a hole transport layer (HTL) aiming to harvest photons in the UV region and emit visible light into the neighboring photoactive layer. Meanwhile, plasmonic grating structures, which were created on the photoactive layer surfaces via the nanoimprinting technique, provided an enhancement effect through light scattering and GCSPR. Thus, an excellent enhancement of OSC efficiency with a significant increase in short circuit photocurrent (J SC) and power conversion efficiency (PCE) in comparison to that of the reference cell was achieved. The fabricated device provides a J SC value as high as 8.41 mA cm-2 (a 14.11% enhancement) and a PCE value of 3.91% (a 19.57% enhancement). The systematic study clearly reveals that the remarkable enhancement of OSC efficiency is achieved by incorporating both AuQD and plasmonic grating.


Contrasting biogeographical patterns in Margarella (Gastropoda: Calliostomatidae: Margarellinae) across the Antarctic Polar Front.
Members of the trochoidean genus Margarella (Calliostomatidae) are broadly distributed across Antarctic and sub-Antarctic ecosystems. Here we used novel mitochondrial and nuclear gene sequences to clarify species boundaries and phylogenetic relationships among seven nominal species distributed on either side of the Antarctic Polar Front (APF). Molecular reconstructions and species-delimitation analyses recognized only four species: M. antarctica (the Antarctic Peninsula), M. achilles (endemic to South Georgia), M. steineni (South Georgia and Crozet Island) and the morphologically variable M. violacea (=M. expansa, M. porcellana and M. pruinosa), with populations in southern South America, the Falkland/Malvinas, Crozet and Kerguelen Islands. Margarella violacea and M. achilles are sister species, closely related to M. steineni, with M. antarctica sister to all these. This taxonomy reflects contrasting biogeographic patterns on either side of the APF in the Southern Ocean. Populations of Margarella north of the APF (M. violacea) showed significant genetic variation but with many shared haplotypes between geographically distant populations. By contrast, populations south of the APF (M. antarctica, M. steineni and M. achilles) exhibited fewer haplotypes and comprised three distinct species, each occurring across a separate geographical range. We hypothesize that the biogeographical differences may be the consequence of the presence north of the APF of buoyant kelps - potential long-distance dispersal vectors for these vetigastropods with benthic-protected development - and their near-absence to the south. Finally, we suggest that the low levels of genetic diversity within higher-latitude Margarella reflect the impact of Quaternary glacial cycles that exterminated local populations during their maxima.


Dietary açai modulates ROS production by neutrophils and gene expression of liver antioxidant enzymes in rats.
Açai (Euterpe oleracea Mart.) has recently emerged as a promising source of natural antioxidants. Because increased oxidative stress and impaired antioxidant defense mechanisms are important factors in the development of diabetic complications and many health claims have been reported for açai, the present study was undertaken to evaluate the possible protective effects of açai on the production of reactive oxygen species by neutrophils and on the liver antioxidant defense system in control and streptozotocin-induced diabetic rats. Diet supplementation with 2% açai was found to increase mRNA levels for gamma-glutamylcysteine synthetase and glutathione peroxidase in liver tissue and to decrease reactive oxygen species production by neutrophils. Compared to control animals, diabetic rats exhibited lower levels of mRNA coding for Zn-superoxide dismutase, glutathione peroxidase and gamma-glutamylcysteine synthetase and higher levels of reactive oxygen species production by neutrophils, thiobarbituric acid-reactive substances and carbonyl proteins in hepatic tissues. Although açai supplementation was not effective in restore gene expression of antioxidant enzymes in diabetic rats, it showed a protective effect, decreasing thiobarbituric acid-reactive substances levels and increasing reduced glutathione content in the liver. These findings suggest that açai can modulate reactive oxygen species production by neutrophils and that it has a significant favorable effect on the liver antioxidant defense system under fisiological conditions of oxidative stress and partially revert deleterious effects of diabetes in the liver.


Morphology and histochemistry of ovarian changes in the field rat, Millardia meltada.
A study has been made of the morphological and histochemical changes of the ovary of the field rat, Millardia meltada during its oestrous cycle and pregnancy. The follicular growth and atresia, ovulation and formation of corpora lutea occur throughout the year except severe winter months (December and January). Fluctuations in the follicular development occur on different days of the oestrous cycle and pregnancy. The granulosa cells show a progressive increase in their size in successive stages of follicle growth. The granulosae of normal follicles show some sparsely scattered lipid bodies which consist of phospholipids. Theca interna cells during follicular growth develop diffuse lipoproteins and lipid droplets consisting of triglycerides, phospholipids and cholesterol and/or its esters. The luteal cells of corpora lutea are formed by the granulosa cells as the theca interna cells degenerate and disappear. The fibroblast-like cells of thecal origin, alongwith the blood vessels, invade the luteal cell mass. The luteal cells during metoestrus, dioestrus and first half of pregnancy show abundant diffuse lipoproteins and a few lipid droplets composed mainly of phospholipids and some triglycerides, which are indicative of active steroidogenesis. The details of degenerative histological and histochemical alterations of corpora lutea during oestrous cycle and pregnancy are also described and discussed. Morphological and histochemical changes of follicular atresia are described. The granulosa cells of atretic follicle degenerate and disappear leaving behind theca interna cells which form patches of interstitial gland cells during the reproductive activity of the present rat. Interstitial gland cells show diffusely distributed sudanophilic lipoproteins and lipid droplets consisting of triglycerides, cholesterol and/or its esters and some phospholipids, which are indicative of steroidogenesis. The functional significance of histological and histochemical changes, which occur in various components of the ovary during oestrous cycle and pregnancy, has been discussed.