From Semantics to Execution: Integrating Action Planning With Reinforcement Learning for Robotic Causal Problem-Solving.
Reinforcement learning is generally accepted to be an appropriate and successful method to learn robot control. Symbolic action planning is useful to resolve causal dependencies and to break a causally complex problem down into a sequence of simpler high-level actions. A problem with the integration of both approaches is that action planning is based on discrete high-level action- and state spaces, whereas reinforcement learning is usually driven by a continuous reward function. Recent advances in model-free reinforcement learning, specifically, universal value function approximators and hindsight experience replay, have focused on goal-independent methods based on sparse rewards that are only given at the end of a rollout, and only if the goal has been fully achieved. In this article, we build on these novel methods to facilitate the integration of action planning with model-free reinforcement learning. Specifically, the paper demonstrates how the reward-sparsity can serve as a bridge between the high-level and low-level state- and action spaces. As a result, we demonstrate that the integrated method is able to solve robotic tasks that involve non-trivial causal dependencies under noisy conditions, exploiting both data and knowledge.


An enzyme linked immunosorbent assay (ELISA) for the determination of the human haptoglobin phenotype.
Haptoglobin (Hp) is an abundant serum protein which binds extracorpuscular hemoglobin (Hb). Two alleles exist in humans for the Hp gene, denoted 1 and 2. Diabetic individuals with the Hp 2-2 genotype are at increased risk of developing vascular complications including heart attack, stroke, and kidney disease. Recent evidence shows that treatment with vitamin E can reduce the risk of diabetic vascular complications by as much as 50% in Hp 2-2 individuals. We sought to develop a rapid and accurate test for Hp phenotype (which is 100% concordant with the three major Hp genotypes) to facilitate widespread diagnostic testing as well as prospective clinical trials. A monoclonal antibody raised against human Hp was shown to distinguish between the three Hp phenotypes in an enzyme linked immunosorbent assay (ELISA). Hp phenotypes obtained in over 8000 patient samples using this ELISA method were compared with those obtained by polyacrylamide gel electrophoresis or the TaqMan PCR method. Our analysis showed that the sensitivity and specificity of the ELISA test for Hp 2-2 phenotype is 99.0% and 98.1%, respectively. The positive predictive value and the negative predictive value for Hp 2-2 phenotype is 97.5% and 99.3%, respectively. Similar results were obtained for Hp 2-1 and Hp 1-1 phenotypes. In addition, the ELISA was determined to be more sensitive and specific than the TaqMan method. The Hp ELISA represents a user-friendly, rapid and highly accurate diagnostic tool for determining Hp phenotypes. This test will greatly facilitate the typing of thousands of samples in ongoing clinical studies.


Nabiximols (THC/CBD oromucosal spray, Sativex®) in clinical practice--results of a multicenter, non-interventional study (MOVE 2) in patients with multiple sclerosis spasticity.
Nabiximols (Sativex®), a cannabinoid-based oromucosal spray, is an add-on therapy for patients with moderate to severe multiple sclerosis spasticity (MSS) resistant to other medications. The primary objective was to provide real-life observational data of clinical experience of nabiximols in contrast to formal clinical trials of effectiveness. This was an observational, prospective, multicenter, non-interventional study with a follow-up period of 3-4 months, conducted in routine care setting in Germany. Patients with moderate to severe MSS were included at nabiximols' initiation. Structured documentation forms, questionnaires and validated instruments were used for data collection at inclusion, 1 and 3 months after inclusion. Overall, 335 patients were assessed of whom 276 fitted the criteria and were included in the effectiveness analysis. After 1 month, nabiximols provided relief of resistant MSS in 74.6% of patients according to specialist assessment; mean spasticity 0-10 numerical rating scale (NRS) score decreased from 6.1 ± 1.8 to 5.2 ± 2.0 points; in patients with NRS improvement ≥20% mean NRS score decreased by 40%. After 3 months, 55.3% of patients had continued to use nabiximols and the mean NRS score had decreased by 25% from baseline. 17% of patients reported adverse events. Real-life data confirm nabiximols as an effective and well-tolerated treatment option for resistant MSS in clinical practice.


Peripheral levels of matrix metalloproteinase-9, interleukin-6, and C-reactive protein are elevated in patients with acute coronary syndromes: correlations with serum troponin I.
Acute coronary syndromes (ACS) are characterized by activation of systemic and local inflammatory mediators. The interrelation between these soluble inflammatory markers and their association with markers of myocardial necrosis have not been extensively studied. The study was undertaken to evaluate the association of the systemic levels of matrix metalloproteinase-9 (MMP-9) and the tissue inhibitor of metalloproteinase-1 (TIMP-1), with C-reactive protein (CRP), interleukin-6 (IL-6), and serum troponin-I in patients admitted with ACS. Analysis of serum concentrations of the above inflammatory markers was performed in 53 patients with unstable angina (UA) and in 15 with non-ST-segment elevation myocardial infarction (NSTEMI) within 48 h of admission, and 34 patients with stable coronary artery disease. Compared with patients with stable angina, those with ACS had elevated admission levels of MMP-9 (p = 0.04), CRP (p < 0.001), and IL-6 (p = 0.001), but not TIMP-1 (p = 0.55). Compared with patients with UA, those with NSTEMI also had higher levels of IL-6 (p < 0.001), CRP (p = 0.002), and MMP-9 (p = 0.05). In patients with ACS, the admission levels of inflammatory mediators, including MMP-9, CRP, and IL-6 are significantly elevated, specifically in association with serum troponin I. Systemic and local markers of inflammatory activity may be directly associated with myocardial injury.


Ancillary Service Utilization and Impact in Huntington's Disease.
Prior Huntington's disease (HD) studies suggest ancillary services improve motor symptoms, cognition, mood, and quality of life but frequency of use and clinicalcharacteristics are unclear. Describe ancillary service utilization in a cohort of individuals with HD and determine which participant characteristics are associated with ancillary service utilization. Retrospective cross-sectional analysis of Enroll-HD database. Participants were grouped by therapy: physical and/or occupational (PT/OT), psychotherapy and/or counseling (PC), speech and/or swallowing (ST). We performed bivariate comparisons analysis of demographic and disease characteristics between those with/without each therapy and to analyze one-year mean change in assessment scores. Of 4751 participants, 1537 (32.35%) utilized therapies (11.82% PT/OT, 5.33% PC, 3.01% ST, 1.98% all three, 10.21% two therapies). PT/OT participants had worse motor and functional scores: mean UHDRS motor score (41.17 vs. 38.05, p = 0.002), median total functional capacity score (TFC) (8.00 vs. 9.00, p < 0.001). PC participants had worse mood but better cognitive and functional scores: median depression score (7.00 vs. 2.00, p < 0.001), median MMSE (28.00 vs. 26.00, p < 0.001), median TFC (10.00 vs. 8.00, p < 0.001). ST participants had more dysarthria, and worse cognitive and functional scores: dysarthria (32.2% vs. 20.1% p < 0.001), mean correct Symbol Digit Modality Test (16.79 vs. 23.27, p < 0.001), median TFC (6.00 vs. 9.00, p < 0.001). Over one year, PC participants' depression scores improved compared to untreated (- 1.24 vs. - 0.11, p = 0.040). ST participants' depression scores worsened (1.14 vs. - 0.23, p = 0.044). Mean change in TFC was not significant for any therapies. Only 32% of Enroll-HD site participants received ancillary services. Use correlated with expected clinical characteristics, though impact of use remains unclear.


[Pyruvate kinase from Calicophoron ijimai trematodes and its potential inhibition by anthelmintic preparations].
It was shown that pyruvate kinase (PK) in the supernatant fraction from Calicophoron ijimai is able to regulate the direction of metabolic flow at glucose break down from phosphoenolpyruvate (PEP) level. The enzyme for activity required substrate, dinucleotides, cations K+ and Mn++. The activity with Mg++ as divalent cation is low. The addition of fructose-1.6-diphosphate (FDP) did not affect the enzyme activity with Mn++, however, increased the affinity for PEP. The velocity of Mg++ activated reaction increased by 8.2 times in the presence of FDP. PK in C. ijimai is sensitive to ATP inhibition, being weakly inhibited by malate. L-alanine did not influence on the enzyme activity. The effect of some anthelminthic preparations on the PK activity was shown.


Reducing bias and variance for CTF estimation in single particle cryo-EM.
When using an electron microscope for imaging of particles embedded in vitreous ice, the recorded image, or micrograph, is a significantly degraded version of the tomographic projection of the sample. Apart from noise, the image is affected by the optical configuration of the microscope. This transformation is typically modeled as a convolution with a point spread function. The Fourier transform of this function, known as the contrast transfer function (CTF), is oscillatory, attenuating and amplifying different frequency bands, and sometimes flipping their signs. High-resolution reconstruction requires this CTF to be accounted for, but as its form depends on experimental parameters, it must first be estimated from the micrograph. We present a new method for CTF estimation based on multitaper techniques that reduce bias and variance in the estimate. We also use known properties of the CTF and the background power spectrum to further reduce the variance through background subtraction and steerable basis projection. We show that the resulting power spectrum estimates better capture the zero-crossings of the CTF and yield accurate CTF estimates on several experimental micrographs.


Copy number variations of neurotrophic tyrosine receptor kinase 3 (NTRK3) may predict prognosis of ovarian cancer.
Platinum resistance is a critical barrier for clinicians to improve the survival of ovarian cancer. Our study evaluated the correlation between copy number variations (CNVs) of neurotrophic tyrosine receptor kinase 3 (NTRK3) and the prognosis of ovarian cancer, which might predict platinum resistance in ovarian cancer patients.Array comparative genomic hybridization (CGH) was used to test gene backgrounds between platinum-sensitive and platinum-resistant relapsed populations and CNVs of NTRK3 were indicated by cluster analysis. Fluorescence in situ hybridization (FISH) was adopted in 41 cases for further verification, which confirmed the results of array CGH. Spearman's rank correlation analysis and χ test were used to evaluate the accuracy of CNVs of NTRK3 which predicted platinum-sensitive or platinum-resistant recurrence.We detected CNVs of NTRK3 between 2 groups by array CGH, and amplification of NTRK3 was confirmed by FISH in the platinum-sensitive recurrence group with enlarged samples. The test concordance of 2 methods was 78.6%. Among 41 cases with satisfied FISH results, the median time to recurrence (TTR) of patients with amplified and nonamplified NTRK3 were respectively 18 and 5 months (P <.01). The cut-off value of TTR to differentiate platinum-sensitive or platinum-resistant recurrence was 6 months in accordance with clinical practice. According to the above standard, 15 cases with NTRK3 amplification were platinum-sensitive and 12 cases without NTRK3 amplification were platinum-resistant recurrences which demonstrated that the accuracy of NTRK3 amplification/nonamplification to predict recurrent types was 65.9% (27/41).CNVs of NTRK3 were associated with platinum-sensitive and platinum-resistant recurrences. Amplification of NTRK3 perfectly predicted platinum-sensitive relapse of ovarian cancer.


Doctors' and nurses' perceptions of ethical problems in end-of-life decisions.
To identify and compare doctors' and nurses' perceptions of ethical problems. Rationale. Ethical problems are a source of tension for health professionals. Misunderstandings or conflicts may result from differing perceptions of ethical problems. If true collaboration is to be achieved, it is important to understand the perspectives of others, particularly when difficult end-of-life decisions must be made. In this qualitative study a total of seven doctors and 14 nurses working in acute care adult medical-surgical areas, including intensive care, were asked to describe ethical problems that they frequently encounter in practice. Interviews were taped and transcribed. Thematic analysis followed. All participants experienced ethical problems around decision making at the end of life. The core problem for both doctors and nurses was witnessing suffering, which engendered a moral obligation to reduce that suffering. Uncertainty about the best course of action for the patient and family was a source of moral distress. Competing values, hierarchical processes, scarce resources, and communication emerged as common themes. The key difference between the groups was that doctors are responsible for making decisions and nurses must live with these decisions. Each group, therefore, asked different questions when encountering and interpreting sources of moral distress. It was concluded that observed differences between doctors and nurses were a function of the professional role played by each rather than differences in ethical reasoning or moral motivation. Although this was a small qualitative study on one institution, and may not be generalizable, results suggest that doctors and nurses need to engage in moral discourse to understand and support the ethical burden carried by the other. Administrators should provide opportunities for discourse to help staff reduce moral distress and generate creative strategies for dealing with this.


A thermosensitive hydrogel formulation of phage and colistin combination for the management of multidrug-resistant Acinetobacter baumannii wound infections.
Chronic skin wounds are often associated with multidrug-resistant bacteria, impeding the healing process. Bacteriophage (phage) therapy has been revitalized as a promising strategy to counter the growing concerns of antibiotic resistance. However, phage monotherapy also faces several application drawbacks, such as a narrow host spectrum, the advent of resistant phenotypes and poor stability of phage preparations. Phage-antibiotic synergistic (PAS) combination therapy has recently been suggested as a possible approach to overcome these shortcomings. In the present study, we employed a model PAS combination containing a vB_AbaM-IME-AB2 phage and colistin to develop stable wound dressings of PAS to mitigate infections associated with Acinetobacter baumannii. A set of thermosensitive hydrogels were synthesized with varying amounts of Pluronic® F-127 (PF-127 at 15, 17.5 and 20 w/w%) modified with/without 3 w/w% hydroxypropyl methylcellulose (HPMC). Most hydrogel formulations had a gelation temperature around skin temperature, suitable for topical application. The solidified gels were capable of releasing the encapsulated phage and colistin in a sustained manner to kill bacteria. The highest bactericidal effect was achieved with the formulation containing 17.5% PF-127 and 3% HPMC (F5), which effectively killed bacteria in both planktonic (by 5.66 log) and biofilm (by 3 log) states and inhibited bacterial regrowth. Good storage stability of F5 was also noted with negligible activity loss after 9 months of storage at 4 °C. The ex vivo antibacterial efficacy of the F5 hydrogel formulation was also investigated in a pork skin wound infection model, where it significantly reduced the bacterial burden by 4.65 log. These positive outcomes warrant its further development as a topical PAS-wound dressing.


Intradomain Confinement of Disulfides in the Folding of Two Consecutive Modules of the LDL Receptor.
The LDL receptor internalizes circulating LDL and VLDL particles for degradation. Its extracellular binding domain contains ten (seven LA and three EGF) cysteine-rich modules, each bearing three disulfide bonds. Despite the enormous number of disulfide combinations possible, LDLR oxidative folding leads to a single native species with 30 unique intradomain disulfides. Previous folding studies of the LDLR have shown that non native disulfides are initially formed that lead to compact species. Accordingly, the folding of the LDLR has been described as a "coordinated nonvectorial" reaction, and it has been proposed that early compaction funnels the reaction toward the native structure. Here we analyze the oxidative folding of LA4 and LA5, the modules critical for ApoE binding, isolated and in the LA45 tandem. Compared to LA5, LA4 folding is slow and inefficient, resembling that of LA5 disease-linked mutants. Without Ca++, it leads to a mixture of many two-disulfide scrambled species and, with Ca++, to the native form plus two three-disulfide intermediates. The folding of the LA45 tandem seems to recapitulate that of the individual repeats. Importantly, although the folding of the LA45 tandem takes place through formation of scrambled isomers, no interdomain disulfides are detected, i.e. the two adjacent modules fold independently without the assistance of interdomain covalent interactions. Reduction of incredibly large disulfide combinatorial spaces, such as that in the LDLR, by intradomain confinement of disulfide bond formation might be also essential for the efficient folding of other homologous disulfide-rich receptors.


Ethnic Identity, Questionnaire Content, and the Dilemma of Race Matching in Surveys of African Americans by African American Interviewers.
We used data from two telephone-administered health surveys to explore African Americans' preferences for interviewer race. The first survey utilized African American interviewers to assess ethnic identity and aspects of healthy eating among 617 African American adults. In the second survey, interviewers of varying races queried 534 African American adults about their motivations to eat healthier. The motivation survey contained almost no racial content, whereas 40% of the ethnic identity survey assessed racial content. Using ethnic identity survey data only, we found that respondents with Afrocentric or Black American identity components were more likely to prefer African American interviewers than respondents with solely Assimilated, Bicultural, or Multicultural identity components. Ethnic identity survey respondents were also more likely to prefer racially/ethnically matched interviewers than motivation survey respondents. Ethnic identity respondents with a college or graduate degree reported lower hypothetical comfort with a White interviewer than respondents with a high school education.


Identification of Legionella pneumophila by latex agglutination.
A latex agglutination test for the identification of Legionella pneumophila serogroups 1 through 6 is described. The reagent is specific for L. pneumophila and enables the ready identification of L. pneumophila colonies on agar plates. Preliminary evidence suggests that latex agglutination enables the detection of soluble L. pneumophila antigens in respiratory secretions of patients suspected of having legionellosis.


Interferon-alpha-induced changes in tryptophan metabolism. relationship to depression and paroxetine treatment.
Tryptophan (TRP) degradation into kynurenine (KYN) by the enzyme, indoleamine-2,3-dioxygenase, during immune activation may contribute to development of depressive symptoms during interferon (IFN)-alpha therapy. Twenty-six patients with malignant melanoma were randomly assigned in double-blind fashion to receive either placebo or paroxetine, beginning 2 weeks before IFN-alpha treatment and continuing for the first 12 weeks of IFN-alpha therapy. At treatment initiation and at 2, 4, and 12 weeks of IFN-alpha treatment, measurements of TRP, KYN, and neopterin (a marker of immune activation), were obtained, along with structured assessments of depression, anxiety, and neurotoxicity. Regardless of antidepressant treatment status, all patients exhibited significant increases in KYN, neopterin, and the KYN/TRP ratio during IFN-alpha therapy. Among antidepressant-free patients, patients who developed major depression exhibited significantly greater increases in KYN and neopterin concentrations and more prolonged decreases in TRP concentrations than did nondepressed, antidepressant-free patients. Moreover, in antidepressant-free patients, decreases in TRP correlated with depressive, anxious, and cognitive symptoms, but not neurovegetative or somatic symptoms. No correlations were found between clinical and biological variables in antidepressant-treated patients. The results suggest that reduced TRP availability plays a role in IFN-alpha-induced depressive symptoms, and paroxetine, although not altering the KYN or neopterin response to IFN-alpha, attenuates the behavioral consequences of IFN-alpha-mediated TRP depletion.


Changes in initial COPD treatment choice over time and factors influencing prescribing decisions in UK primary care: in UK primary care: a real-world, retrospective, observational.
Prescribing patterns in chronic obstructive pulmonary disease (COPD) are often inconsistent with published guidelines. This retrospective, observational study utilised data from the Optimum Patient Care Research Database to examine the changes in COPD prescribing patterns over time and to identify predictors of physician treatment choice for patients newly diagnosed with COPD. Initial therapy was defined as the treatment(s) prescribed at or within 1 year before COPD diagnosis. Changes over time were assessed in three cohorts based on the date of diagnosis: (1) 1997-2001; (2) 2002-2006; and (3) 2007-2010. Factors affecting the odds of being prescribed any initial therapy or any initial maintenance therapy were identified by univariable and multivariable logistic regression. The analysis included 20,154 patients, 45% of whom were prescribed an initial regimen containing an inhaled corticosteroid (ICS), whereas 28% received no initial pharmacological treatment. Prescribing of ICS monotherapy decreased over time, as did the proportion of patients receiving no therapy at or within 1 year before diagnosis. Comorbid asthma, a high exacerbation rate, increased symptoms and poor lung function each increased the likelihood of being prescribed any initial therapy or initial maintenance therapy; comorbid asthma and an annual rate of ⩾3 exacerbations were the strongest predictors. In conclusion, our analyses revealed major differences between actual prescribing behaviour and guideline recommendations for patients with newly diagnosed COPD, with many patients receiving no treatment and large numbers of patients receiving ICS-containing regimens. Predictors of initial therapy were identified.


[Epidermoid carcinoma of the penis].
Four cases of Penis Epidermoid Carcinoma, three treated with partial amputation and one with Co60 radiotherapy due to the patient's refusal to undergo surgery, are illustrated. After an average follow-up of 14 months (2-24), patients who underwent surgery have not relapsed. Local relapse was observed in the patient given radiotherapy within 6 months.


Robotic-assisted gait training in Parkinson's disease: a three-month follow-up randomized clinical trial.
The aim of this study was to evaluate the efficacy of a robotic-assisted gait training (RAGT), together with a conventional exercise program (CEP), to improve PD ambulation, as compared to standard gait training. Thirty-eight patients with mild PD stage (H&Y 2-2.5) were randomly assigned to an experimental group (EG) or a control group. The 19 patients in EG received 30 min RAGT (using Lokomat device), whereas the 19 controls received a conventional gait training; both groups received 30 min of CEP. Participants were evaluated before (T0), immediately after (T1), and 12 weeks after the end of treatment (T2), by using 10-MWT, Tinetti Test and the motor score of the UPDRS-III. We found that Tinetti Walking (TW) (X2(3) = 31.75; p < 0.001), Tinetti Balance (X2(3) = 74.07; p < 0.001), UPDRS-III (X2(3) = 6.87; p < 0.001) and GDS (X2(3) = 28.83; p < 0.001) scores were affected by the type of the rehabilitative treatment. At T2, we found a significant difference between the two groups for TW (t = 2.62; p < 0.02, d = 0.85). Concerning all the study outcomes, a significant improvement was observed from T0 to T1 in both groups. However, the functional motor gain at T2 was maintained only in the EG. RAGT may significantly improve walking ability, motor function and for a maximum period of three months. Thus, our findings support the importance of a RAGT as a valid rehabilitative tool for PD.


Monitoring Pc 4 photodynamic therapy in clinical trials of cutaneous T-cell lymphoma using noninvasive spectroscopy.
Silicon phthalocyanine Pc 4 photodynamic therapy (Pc 4-PDT) has emerged as a potentially effective treatment for cutaneous T-cell lymphoma (CTCL). Noninvasive reflectance and fluorescence spectroscopy before, during, and after PDT may provide useful dose metrics and enable therapy to be tailored to individual lesions. We present the design and implementation of a portable bedside spectroscopy system for initial clinical trials of Pc 4-PDT of CTCL. Reflectance and fluorescence spectra were obtained from an early stage CTCL patient throughout the course of the PDT treatment. Preliminary patient data show a significant effect of Pc 4 on the tissue absorption, modest Pc 4 photobleaching, and heterogeneity of Pc 4 within and between the lesions.


Characterization of melanogenesis and morphogenesis of melanosomes by physicochemical properties of melanin and melanosomes in malignant melanoma.
This study elucidates the nature of melanogenesis in B16 and Harding-Passey (HP) mouse melanomas producing melanin and melanosomes of different color and fine structure, i.e., brown-black eumelanosome-like B16 granules and reddish brown pheomelanosome-like HP granules, and compares them with "typical" 3,4-dihydroxyphenylalanine (DOPA) and sepia eumelanins and sepia eumelanosomes. The melanin content of B16 melanosomes was more than three times higher than that of HP melanosomes. The content of free and protein-bound DOPA and 5-S-cysteinyldopa varied greatly in B16, HP, and sepia melanosomes and was unrelated to melanin content. Chemical analysis of the eumelanin: pheomelanin ratio in melanosomes and elemental analysis of isolated melanin showed that B16 and HP melanins are primarily eumelanic, with a higher ratio of pheomelanic component in HP melanin. The spectra of electron spin resonance and IR and X-ray small-angle scattering of B16 and HP melanins were basically similar to those of sepia and DOPA melanins. B16, HP, and DOPA melanins were dissolved in aqueous NH3, while sepia melanin was dissolved to a far lesser extent. It was concluded that both B16 and HP melanomas are primarily involved in eumelanogenesis, although the fine structure of their melanosomes is entirely different, and that the marked color difference in the two melanosomes is related to a difference in the absolute content of eumelanin, the presence of a small amount of pheomelanin, and the mode of chemical bindings of melanin to structural proteins. In contrast to normal skin and hair, melanosome morphogenesis may not directly correspond to melanogenesis type in malignant melanoma.


Acute phenytoin and primidone intoxication: a pharmacokinetic analysis.
Serial plasma levels of phenytoin, primidone, and phenobarbital were determined in a patient following massive overdose of phenytoin and primidone. The patient's neurologic status improved slowly over a period of 10 days and correlated best with the rise and fall of phenytoin plasma concentrations. The pharmacokinetics of all three agents were characterized by nonlinear regression analysis of their respective plasma concentration-time profiles during the elimination phase, followed by analog computer simulations of their entire plasma concentration-time profile closely resembled the observed values. Average values of Km and Vmax obtained from patients undergoing chronic therapy were used in the simulation and appear to adequately describe phenytoin elimination in this overdose situation. The elimination half-lives of primidone and phenobarbital of 6.2 and 83.5 hours, respectively, were within the "normal range" for patients on chronic therapy. Two distinct absorption phases for primidone and three for phenytoin were noted. The marked decrease in the estimated absorption rate constant between phases 1 and 2 for each drug may have been due to slow dissolution of a large congealed mass of phenytoin and primidone in the gut. The analysis of serial plasma samples following a massive overdose is recommended to provide a reliable data base for therapeutic decisions.


Etiology of patent ductus arteriosus in dogs.
Patent ductus arteriosus (PDA) is the most common congenital heart disease in dogs and usually causes heart failure and death unless corrected at a young age. Previous histologic studies in a line of dogs derived from Miniature Poodles with hereditary PDA identified varying degrees of hypoplasia and asymmetry of ductus-specific smooth muscle and the presence of aortalike elastic tissue in the ductus wall sufficient to cause patency. To determine if similar structural abnormalities cause PDA in other dogs, serial-section, 3-dimensional histology of ductal architecture was studied in 8 non-Poodle purebred dogs with PDA with no immediate family history of PDA. Morphologic abnormalities were observed in 7 of 8 dogs with PDA and essentially were the same as those in dogs known to have a hereditary form of PDA. These findings suggest that apparently sporadic PDA in these breeds is caused by a genetic defect in the structure of the ductus arteriosus that is similar or identical to that in the Poodle. The relatives of dogs with PDA, particularly parents, offspring, and siblings, should be screened for evidence of PDA. Dogs with PDA should not be used for breeding, regardless of breed.


Role of uricase in the triggering of germination of Bacillus fastidiosus spores.
The likelihood that uric acid was the only compound capable of triggering germination of Bacillus fastidiosus spores was reinforced by the finding that ureidoglycollic acid, urea, NH4Cl, 2,8-dihydroxypurine and a combination of L-alanine and O-carbamoyl-D-serine were ineffective as germinants. Uric acid-triggered germination of B. fastidiosus was prevented by a range of inhibitors that also inhibited uricase activity in dormant spore extracts. O2 uptake during germination started immediately after addition of uric acid, possibly as a consequence of the oxidation of uric acid by the enzyme uricase. Germination showed a dependence on uric acid concentration, with a relatively high Km (4-5 mM). During the first 10 min of germination of heat-activated spores there was no detectable change in the number of spore-cortex reducing groups, indicating that selective cortex hydrolysis is not involved in the trigger mechanism of germination of B. fastidiosus. On the basis of the results, a model is proposed in which re-initiation of uricase activity is the mechanism by which B. fastidiosus spores are triggered to emerge from the dormant state.


Impact of vitamin B12 on rhamnose metabolism, stress defense and in-vitro virulence of Listeria monocytogenes.
Listeria monocytogenes is a facultative anaerobe which can cause a severe food-borne infection known as listeriosis. L. monocytogenes is capable of utilizing various nutrient sources including rhamnose, a naturally occurring deoxy sugar abundant in foods. L. monocytogenes can degrade rhamnose into lactate, acetate and 1,2-propanediol. Our previous study showed that addition of vitamin B12 stimulated anaerobic growth of L. monocytogenes on rhamnose due to the activation of bacterial microcompartments for 1,2-propanediol utilization (pdu BMC) with concomitant production of propionate and propanol. Notably, anaerobic 1,2-propanediol metabolism has been linked to virulence of enteric pathogens including Salmonella spp. and L. monocytogenes. In this study we investigated the impact of B12 and BMC activation on i) aerobic and anerobic growth of L. monocytogenes on rhamnose and ii) the level of virulence. We observed B12-induced pdu BMC activation and growth stimulation only in anaerobically grown cells. Comparative Caco-2 virulence assays showed that these pdu BMC-induced cells have significantly higher translocation efficiency compared to non-induced cells (anaerobic growth without B12; aerobic growth with or without B12), while adhesion and invasion capacity is similar for all cells. Comparative proteome analysis showed specific and overlapping responses linked to metabolic shifts, activation of stress defense proteins and virulence factors, with RNA polymerase sigma factor SigL, teichoic acid export ATP-binding protein TagH, DNA repair and protection proteins, RadA and DPS, and glutathione synthase GshAB, previously linked to activation of virulence response in L. monocytogenes, uniquely upregulated in anaerobically rhamnose grown pdu-induced cells. Our results shed light on possible effects of B12 on L. monocytogenes competitive fitness and virulence activation when utilizing rhamnose in anaerobic conditions encountered during transmission and the human intestine.


Multiple roles of the PI3K/PKB (Akt) pathway in cell cycle progression.
As its role in tumor progression emerges, the PI3K/PKB (Akt) pathway presents an appealing cancer therapeutic target. Recent studies have investigated the mechanisms underlying the tumor-promoting effects of this pathway. PKB triggers a network that positively regulates G1/S cell cycle progression through inactivation of GSK3-beta, leading to increased cyclin D1, and inhibition of Forkhead family transcription factors and the tumor suppressor tuberin (TSC2), leading to reduction of p27Kip1. The identification of p21Waf1/Cip1 and p27Kip1 as novel substrates of PKB provided new insights into mechanisms whereby hyperactivation of this lipid signaling pathway may lead to cell cycle deregulation in human cancers. The PI3K pathway may also play a key role in the G2/M transition and its constitutive activation may lead to defects in DNA damage checkpoint control.


[The results of phylogenetic and experimental research into the etiology of pemphigus vulgaris].
An antigen, binding with antibodies from patients with pemphigus vulgaris, has been found in the epidermis of a shedding Natrix natrix. This permitted a hypothesis on the atavistic nature of pemphigus, consisting in the following: abolition of tolerance to pemphigus antigen (PA) in humans and mammals is explained by the release of the 'shedding' atavistic gene, that codes for the humoral factor responsible for PA expression in keratinocytes. This hypothesis has been confirmed by experimental induction of pemphigus-like changes of the serpentine and murine epidermis (by subcutaneous injections of a shedding snake serum protein--SSSP), as well as by experiments with SSSP blocking of pemphigus induced by passive transfer of antibodies from patients with pemphigus vulgaris to newborn BALB/s mice.


Ewald sums for one dimension.
We derive analytic solutions for the potential and field in a one-dimensional system of masses or charges with periodic boundary conditions, in other words, Ewald sums for one dimension. We also provide a set of tools for exploring the system evolution and show that it is possible to construct an efficient algorithm for carrying out simulations. In the cosmological setting we show that two approaches for satisfying periodic boundary conditions-one overly specified and the other completely general-provide a nearly identical clustering evolution until the number of clusters becomes small, at which time the influence of any size-dependent boundary cannot be ignored. Finally, we compare the results with other recent work with the hope to provide clarification over differences these issues have induced.


Personalizing Environmental Health: At the Intersection of Precision Medicine and Occupational Health in the Military.
: Recent efforts in precision medicine present unique opportunities for military environmental and occupational health. Risk assessments can be refined by individualized risk factors such as genomics, and health status can be monitored and informed using mobile health (mHealth) devices. The military currently monitors exposures with service-wide databases and has one of the world's largest biobanks of serum samples available for health surveillance. New approaches are being developed for risk assessment, novel exposure-based biomarkers, and mobile applications to combine the facile collection of exposure data with tracking and planning utility. Planning by military leaders and coordination with national efforts puts the Department of Defense (DoD) in a unique position to benefit both Service Members and the nation, as reviewed in a symposium cosponsored by the DoD and the Johns Hopkins University-Applied Physics Laboratory (October 27 to 28, 2015).


Auditory evoked potentials to spectro-temporal modulation of complex tones in normal subjects and patients with severe brain injury.
In order to assess higher auditory processing capabilities, long-latency auditory evoked potentials (AEPs) were recorded to synthesized musical instrument tones in 22 post-comatose patients with severe brain injury causing variably attenuated behavioural responsiveness. On the basis of normative studies, three different types of spectro-temporal modulation were employed. When a continuous 'clarinet' tone changes pitch once every few seconds, N1/P2 potentials are evoked at latencies of approximately 90 and 180 ms, respectively. Their distribution in the fronto-central region is consistent with generators in the supratemporal cortex of both hemispheres. When the pitch is modulated at a much faster rate ( approximately 16 changes/s), responses to each change are virtually abolished but potentials with similar distribution are still elicited by changing the timbre (e.g. 'clarinet' to 'oboe') every few seconds. These responses appear to represent the cortical processes concerned with spectral pattern analysis and the grouping of frequency components to form sound 'objects'. Following a period of 16/s oscillation between two pitches, a more anteriorly distributed negativity is evoked on resumption of a steady pitch. Various lines of evidence suggest that this is probably equivalent to the 'mismatch negativity' (MMN), reflecting a pre-perceptual, memory-based process for detection of change in spectro-temporal sound patterns. This method requires no off-line subtraction of AEPs evoked by the onset of a tone, and the MMN is produced rapidly and robustly with considerably larger amplitude (usually >5 microV) than that to discontinuous pure tones. In the brain-injured patients, the presence of AEPs to two or more complex tone stimuli (in the combined assessment of two authors who were 'blind' to the clinical and behavioural data) was significantly associated with the demonstrable possession of discriminative hearing (the ability to respond differentially to verbal commands, in the assessment of a further author who was blind to the AEP findings). Behavioural and electrophysiological findings were in accordance in 18/22 patients, but no AEPs could be recorded in two patients who had clear behavioural evidence of discriminative hearing. The absence of long-latency AEPs should not, therefore, be considered indicative of complete functional deafness. Conversely, AEPs were substantially preserved in two patients without behavioural evidence of discriminative hearing. Although not necessarily indicative of conscious 'awareness', such AEP preservation might help to identify sentient patients who are prevented by severe motor disability from communicating their perception.


Inhibition of microsomal prostaglandin E-synthase-1 (mPGES-1) selectively suppresses PGE2 in an in vitro equine inflammation model.
Inhibition of prostaglandin E2 (PGE2) production effectively limits inflammation in horses, however nonspecific prostaglandin blockade via cyclooxygenase (COX) inhibition elicits deleterious gastrointestinal side effects in equine patients. Thus, more selective PGE2 targeting therapeutics are needed to treat inflammatory disease in horses. One potential target is microsomal prostaglandin E-synthase-1 (mPGES-1), which is the terminal enzyme downstream of COX-2 in the inducible PGE2 synthesis cascade. This enzyme has yet to be studied in equine leukocytes, which play a pivotal role in equine inflammatory disease. The objective of this study was to determine if mPGES-1 is a PGE2-selective anti-inflammatory target in equine leukocytes. To evaluate this objective, leukocyte-rich plasma (LRP) was isolated from equine whole blood collected via jugular venipuncture of six healthy adult horses of mixed breeds and genders. LRP was primed with granulocyte-monocyte colony-stimulating factor (GM-CSF) and stimulated with lipopolysaccharide (LPS) in the presence or absence of an mPGES-1 inhibitor (MF63), a COX-2 inhibitor (NS-398), or a nonselective COX inhibitor (indomethacin). Following treatment, mPGES-1 and COX-2 mRNA and protein levels were measured via qPCR and western blot, respectively, and PGE2, thromboxane (TXA2) and prostacyclin (PGI2) levels were measured in cellular supernatants via ELISA. This study revealed that LPS significantly increased mPGES-1 mRNA, but not protein levels in equine LRP as measured by qPCR and western blot, respectively. In contrast, COX-2 mRNA and protein were coordinately induced by LPS. Importantly, treatment of LPS-stimulated leukocytes with indomethacin and NS-398 significantly reduced extracellular concentrations of multiple prostanoids (PGE2, TXA2 and PGI2), while the mPGES-1 inhibitor MF63 selectively inhibited PGE2 production only. mPGES-1 inhibition also preserved higher basal levels of PGE2 production when compared to either COX inhibitor, which might be beneficial in a clinical setting. In conclusion, this work identifies mPGES-1 as a key regulator of PGE2 production and a PGE2-selective target in equine leukocytes. This study demonstrates that mPGES-1 is a potentially safer and effective therapeutic target for treatment of equine inflammatory disease when compared to traditional non-steroidal anti-inflammatory drugs.


Super-Resolution Optical Lithography with DNA.
We developed an efficient, versatile, and accessible super-resolution microscopy method to construct a nanoparticle assembly at a spatial resolution below the optical diffraction limit. The method utilizes DNA and a photoactivated DNA cross-linker. Super-resolution optical techniques have been used only as a means to make measurements below the light diffraction limit. Furthermore, no optical technique is currently available to construct nanoparticle assemblies with a precisely designed shape and internal structure at a resolution of a few tens of nanometers (nm). Here we demonstrate that we can fulfill this deficiency by utilizing spontaneous structural dynamics of DNA hairpins combined with single-molecule fluorescence resonance energy transfer (smFRET) microscopy and a photoactivated DNA cross-linker. The stochastic fluorescence blinking due to the spontaneous folding and unfolding motions of DNA hairpins enables us to precisely localize a folded hairpin and solidify it only when it is within a predesigned target area whose size is below the diffraction limit. As the method is based on an optical microscope and an easily clickable DNA cross-linking reagent, it will provide an efficient means to create large nanoparticle assemblies with a shape and internal structure at an optical super-resolution, opening a wide window of opportunities toward investigating their photophysical and optoelectronic properties and developing novel devices.


An Emerging Human Parechovirus Type 5 Causing Sepsis-Like Illness in Infants in Australia.
Human parechovirus (HPeV), particularly type 3 (HPeV3), is an important cause of sepsis-/meningitis-like illness in young infants. Laboratory records identified a total of ten HPeV-positive cases in Southeastern Australia between January and July 2019. The HPeV present in these cases were typed by Sanger sequencing of the partial viral capsid protein 1 (VP1) region and selected cases were further characterised by additional Sanger or Ion Torrent near-full length virus sequencing. In seven of the ten cases, an HPeV type 5 (HPeV5) was identified, and in the remaining three cases, an HPeV type 1 was identified. The HPeV5-positive cases were infants under the age of 3 months admitted to hospital with fever, rash, lethargy and/or sepsis-like clinical signs. Near full-length virus sequencing revealed that the HPeV5 was most likely a recombinant virus, with structural genes most similar to an HPeV5 from Belarus in 2018, and a polymerase gene most similar to an HPeV3 from Australia in 2013/14. While HPeV5 is not typically associated with severe clinical signs, the HPeV5 identified here may have been able to cause more severe disease in young infants through the acquisition of genes from a more virulent HPeV.


[Registries for implants and new interventions].
An increasing number of registries are being set up in healthcare. Their development is sometimes guided by an explicit desire to control the unknown, as with most registries initiated for new devices as part of coverage with evidence development schemes. By definition, registries cannot answer questions about the effectiveness of interventions. At best, they document the outcomes after the intervention; it will remain unknown what the outcome would have been with a different implant or device, or without one. Nonetheless, registries can provide useful information for other purposes: for documenting appropriate use, for monitoring complications and revisions, for comparing observed outcomes against predicted outcomes, and for comparing different types of devices used for the same purpose. Registries can also assist quality improvement efforts by participating professionals and healthcare organisations.


Kinetics of hydrogen ion diffusion across phospholipid vesicle membranes.
The membrane-impermeant, pH-sensitive fluorescence probe 8-hydroxy-1,3,6-pyrenetrisulfonate can be entrapped within the internal aqueous compartment of unilamellar phospholipid vesicles, where it serves as a reliable indicator of internal aqueous hydrogen ion concentration [Clement, N. R., & Gould, J. M. (1981) Biochemistry 20, 1534-1539]. When the external (medium) pH of a suspension of soybean phospholipid vesicles was rapidly changed from 8.2 to 6.65, the rate of subsequent H+ influx into the vesicles, measured as the change in pyranine fluorescence, was limited (in KCl media) by the rate of charge-compensating counterion redistributions. The half-time for the pyranine fluorescence change (corresponding to an internal pH change from 8.2 to 7.43), which was several minutes in the absence of valinomycin, could be decreased to approximately 300 ms, but not further, by the K+ ionophore valinomycin. Proton ionophores such as gramicidin or bis(hexafluoroacetonyl)acetone (1799), on the other hand, decreased the time required for transmembrane H+ equilibration to less than 1 ms. These findings indicate that the intrinsic permeability of unilamellar vesicle membranes to hydrogen ions is surprisingly high and much greater than the observed permeabilities of other small ions.


Three-dimensional characterization of the vascular bed in bone metastasis of the rat by microcomputed tomography (MicroCT).
Angiogenesis contributes to proliferation and metastatic dissemination of cancer cells. Anatomy of blood vessels in tumors has been characterized with 2D techniques (histology or angiography). They are not fully representative of the trajectories of vessels throughout the tissues and are not adapted to analyze changes occurring inside the bone marrow cavities. We have characterized the vasculature of bone metastases in 3D at different times of evolution of the disease. Metastases were induced in the femur of Wistar rats by a local injection of Walker 256/B cells. Microfil®, (a silicone-based polymer) was injected at euthanasia in the aorta 12, 19 and 26 days after injection of tumor cells. Undecalcified bones (containing the radio opaque vascular casts) were analyzed by microCT, and a first 3D model was reconstructed. Bones were then decalcified and reanalyzed by microCT; a second model (comprising only the vessels) was obtained and overimposed on the former, thus providing a clear visualization of vessel trajectories in the invaded metaphysic allowing quantitative evaluation of the vascular volume and vessel diameter. Histological analysis of the marrow was possible on the decalcified specimens. Walker 256/B cells induced a marked osteolysis with cortical perforations. The metaphysis of invaded bones became progressively hypervascular. New vessels replaced the major central medullar artery coming from the diaphyseal shaft. They sprouted from the periosteum and extended into the metastatic area. The newly formed vessels were irregular in diameter, tortuous with a disorganized architecture. A quantitative analysis of vascular volume indicated that neoangiogenesis increased with the development of the tumor with the appearance of vessels with a larger diameter. This new method evidenced the tumor angiogenesis in 3D at different development times of the metastasis growth. Bone and the vascular bed can be identified by a double reconstruction and allowed a quantitative evaluation of angiogenesis upon time.


The lambda red proteins promote efficient recombination between diverged sequences: implications for bacteriophage genome mosaicism.
Genome mosaicism in temperate bacterial viruses (bacteriophages) is so great that it obscures their phylogeny at the genome level. However, the precise molecular processes underlying this mosaicism are unknown. Illegitimate recombination has been proposed, but homeologous recombination could also be at play. To test this, we have measured the efficiency of homeologous recombination between diverged oxa gene pairs inserted into lambda. High yields of recombinants between 22% diverged genes have been obtained when the virus Red Gam pathway was active, and 100 fold less when the host Escherichia coli RecABCD pathway was active. The recombination editing proteins, MutS and UvrD, showed only marginal effects on lambda recombination. Thus, escape from host editing contributes to the high proficiency of virus recombination. Moreover, our bioinformatics study suggests that homeologous recombination between similar lambdoid viruses has created part of their mosaicism. We therefore propose that the remarkable propensity of the lambda-encoded Red and Gam proteins to recombine diverged DNA is effectively contributing to mosaicism, and more generally, that a correlation may exist between virus genome mosaicism and the presence of Red/Gam-like systems.


A simple fluorescence method for the study of the internalization of particles in cultured cells: Application to asbestos and glass fibres.
A model for the study of internalization of particles in mammalian cells was applied to asbestos and glass fibres. Briefly, a fluorescent fluid-phase endocytic marker, Lucifer Yellow CH (LY), was allowed to be incorporated into the lysosomal compartment of Syrian hamster embryo cells. Mineral fibres that were internalized by the cells subsequently became 'fluorescent', presumably when the fibre-containing endosome fused with the LY-containing lysosomes. This method was compared with differential interference contrast (DIC) optics. Approximately three times as many of the cell-associated fibres were determined to be internalized by the fluorescence method compared with DIC optics. Both fine and coarse glass fibres were internalized as effectively as asbestos fibres. The relative frequency of internalized (i.e. fluorescent) fibres increased until 4 hr after exposure compared with the total number of cell-associated fibres. The frequency of internalized fibres compared with the number of cell-associated fibres was constant over the range of fibre levels studied. A surface modification (octadecyldimethylchlorosilane-derivatization) of amosite fibres that decreased the carcinogenicity of the fibres, decreased slightly the number of internalized fibres relative to the number of cell-associated fibres, but this was not statistically significant. Cytoskeleton-interfering agents significantly decreased the relative number of internalized fibres.


Predation by Coccinella septempunctata and Harmonia axyridis (Coleoptera: Coccinellidae) on Aphis glycines (Homoptera: Aphididae).
Coccinella septempunctata L. and Harmonia axyridis Pallas are key natural enemies of soybean aphid, Aphis glycines Matsumura, in North America. Third instars, adult females, and adult males of both C. septempunctata and H. axyridis exhibited a type II functional response for predation toward adult soybean aphids at 26 +/- 1 degrees C. In C. septempunctata, the functional response curve of adult males differed from those of third instars and adult females, but there was no difference between third instars and adult females. In H. axyridis, the functional response curves of larvae, adult females, and adult males all differed significantly. Third instars and adult females consumed significantly more soybean aphids than did adult males at prey densities of 150 and 180 aphids per arena for C. septempunctata and at prey densities of 60, 90, 120, 150, and 180 aphids per arena for H. axyridis. The theoretical maximum daily predation rate of adult aphids by C. septempunctata was predicted to be 204 per third instar, 277 per adult female, and 166 per adult male, and 244, 156, and 73, respectively, for H. axyridis. Third instars and adult females of both species consumed significantly more aphids than did adult males on soybean plants with the recommended action threshold of 250 soybean aphids per plant. Both C. septempunctata and H. axyridis have high predation capacities and are important in suppressing soybean aphid populations.


Motor Ingredients Derived from a Wearable Sensor-Based Virtual Reality System for Frozen Shoulder Rehabilitation.
Objective. This study aims to extract motor ingredients through data mining from wearable sensors in a virtual reality goal-directed shoulder rehabilitation (GDSR) system and to examine their effects toward clinical assessment. Design. A single-group before/after comparison. Setting. Outpatient research hospital. Subjects. 16 patients with frozen shoulder. Interventions. The rehabilitation treatment involved GDSR exercises, hot pack, and interferential therapy. All patients first received hot pack and interferential therapy on the shoulder joints before engaging in the exercises. The GDSR exercise sessions were 40 minutes twice a week for 4 weeks. Main Measures. Clinical assessments included Constant and Murley score, range of motion of the shoulder, and muscle strength of upper arm as main measures. Motor indices from sensor data and task performance were measured as secondary measures. Results. The pre- and posttest results for task performance, motor indices, and the clinical assessments indicated significant improvement for the majority of the assessed items. Correlation analysis between the task performance and clinical assessments revealed significant correlations among a number of items. Stepwise regression analysis showed that task performance effectively predicted the results of several clinical assessment items. Conclusions. The motor ingredients derived from the wearable sensor and task performance are applicable and adequate to examine and predict clinical improvement after GDSR training.


E-cigarette Marketing Exposure and Subsequent Experimentation Among Youth and Young Adults.
Electronic cigarette (e-cigarette) use has become increasingly prevalent among US youth and young adults in recent years. Exposure to e-cigarette marketing may stimulate e-cigarette use. In this study, we estimated the longitudinal association between e-cigarette marketing exposure and e-cigarette experimentation among US youth and young adult never tobacco users. The analysis included nationally representative samples of youth (ages 12-17; n = 8121) and young adult (ages 18-24; n = 1683) never tobacco users from wave 2 (2014-2015) and wave 3 (2015-2016) of the Population Assessment of Tobacco and Health Study. In the study, researchers measured past-month exposure to e-cigarette marketing through various places (eg, Web sites and events) at wave 2 and e-cigarette experimentation at wave 3. Statistical analysis included multivariable regressions to examine the associations between wave 2 e-cigarette marketing exposure and wave 3 e-cigarette experimentation. At wave 2, 70.7% of youth and 73.9% of young adult never tobacco users reported past-month exposure to e-cigarette marketing; at wave 3, 4.9% and 4.5% of youth and young adults experimented with e-cigarettes, respectively. Youth and young adults exposed to e-cigarette marketing at wave 2 were more likely (adjusted odds ratio = 1.53, 95% confidence interval = 1.07-2.17; and adjusted odds ratio = 2.73, 95% confidence interval = 1.16-6.42, respectively) to have experimented with e-cigarettes at wave 3 than those not exposed. Marketing exposure through each place at wave 2 was associated with e-cigarette experimentation at wave 3. E-cigarette marketing exposure predicted subsequent e-cigarette experimentation among youth and young adult never tobacco users. Increased restrictions on marketing through various channels may help minimize their exposure to e-cigarette marketing messages.


In Vitro and In Vivo Anticancer Activity of the Most Cytotoxic Fraction of Pistachio Hull Extract in Breast Cancer.
Pistacia (Pistacia vera) hulls (PV) is a health product that has been determined to contain bioactive phytochemicals which have fundamental importance for biomedical use. In this study, PV ethyl acetate extraction (PV-EA) fractions were evaluated with the use of an MTT assay to find the most cytotoxic fraction, which was found to be F13b1/PV-EA. After that, HPTLC was used for identify the most active compounds. The antioxidant activity was analyzed with DPPH and ABTS tests. Apoptosis induction in MCF-7 cells by F13b1/PV-EA was validated via flow cytometry analysis and a distinctive nuclear staining method. The representation of genes like Caspase 3, Caspase 8, Bax, Bcl-2, CAT and SOD was assessed via a reverse transcription (RT_PCR) method. Inhabitation of Tubo breast cancer cell development was examined in the BALB-neuT mouse with histopathology observations. The most abundant active components available in our extract were gallic acid and the flavonoid quercetin. The F13b1/PV-EA has antiradical activity evidence by its inhibition of ABTS and DPPH free radicals. F13b1/PV-EA displayed against MCF-7 a suppressive effect with an IC50 value of 15.2 ± 1.35 µg/mL. Also, the expression of CAT, SOD, Caspase 3, Caspase 8 and Bax increased and the expression of Bcl-2 decreased. F13b1/PV-EA dose-dependently inhibited tumor development in cancer-induced mice. Thus, this finding introduces F13b1/PV-EA as an effectual apoptosis and antitumor active agent against breast cancer.


Pharmacotherapeutic management of co-morbid alcohol and opioid use.
Opioid use disorder (OUD) and alcohol use disorder (AUD) are two highly prevalent substance-related disorders worldwide. Co-use of the substances is also quite prevalent, yet there are no pharmacological treatment approaches specifically designed to treat co-morbid OUD and AUD. Here, the authors critically summarize OUD, AUD and opioid/alcohol co-use and their current pharmacotherapies for treatment. They also review the mechanisms of action of opioids and alcohol within the brain reward circuitry and discuss potential combined mechanisms of action and resulting neuroadaptations. Pharmacotherapies that aim to treat AUD or OUD that may be beneficial in the treatment of co-use are also highlighted. Preclinical models assessing alcohol and opioid co-use remain sparse. Lasting neuroadaptations in brain reward circuits caused by co-use of alcohol and opioids remains largely understudied. In order to fully understand the neurobiological underpinnings of alcohol and opioid co-use and develop efficacious pharmacotherapies, the preclinical field must expand its current experimental paradigms of 'single drug' use to encompass polysubstance use. Such studies will provide insights on the neural alterations induced by opioid and alcohol co-use, and may help develop novel pharmacotherapies for individuals with co-occurring alcohol and opioid use disorders.


Sparse PCA with Oracle Property.
In this paper, we study the estimation of the k-dimensional sparse principal subspace of covariance matrix Σ in the high-dimensional setting. We aim to recover the oracle principal subspace solution, i.e., the principal subspace estimator obtained assuming the true support is known a priori. To this end, we propose a family of estimators based on the semidefinite relaxation of sparse PCA with novel regularizations. In particular, under a weak assumption on the magnitude of the population projection matrix, one estimator within this family exactly recovers the true support with high probability, has exact rank-k, and attains a [Formula: see text] statistical rate of convergence with s being the subspace sparsity level and n the sample size. Compared to existing support recovery results for sparse PCA, our approach does not hinge on the spiked covariance model or the limited correlation condition. As a complement to the first estimator that enjoys the oracle property, we prove that, another estimator within the family achieves a sharper statistical rate of convergence than the standard semidefinite relaxation of sparse PCA, even when the previous assumption on the magnitude of the projection matrix is violated. We validate the theoretical results by numerical experiments on synthetic datasets.


MMP-9 inhibition alleviates postoperative cognitive dysfunction by improving glymphatic function via regulating AQP4 polarity.
Postoperative cognitive dysfunction (POCD) is a common complication after surgery, characterized by deficits in memory, attention and cognitive flexibility. However, the underlying mechanisms of POCD remain unclear. Neuroinflammation and blood-brain barrier disruption have been implicated as potential pathological processes. This study explores the neuroprotective effects and mechanisms of the matrix metalloproteinase(MMP-9)inhibitor GM6001 against POCD. We hypothesize GM6001 may reduce neuroinflammation and preserve blood-brain barrier integrity through direct inhibition of MMP-9. Moreover, GM6001 may stabilize aquaporin-4 polarity and glymphatic clearance function by modulating MMP-9-mediated cleavage of dystroglycan, a key protein for aquaporin-4 anchoring. Our results demonstrate GM6001 alleviates postoperative cognitive deficits and neuroinflammation. GM6001 also preserves blood-brain barrier integrity and rescues aquaporin-4 mislocalization after surgery. This study reveals a novel dual role for MMP-9 inhibition in cognitive protection through direct anti-neuroinflammatory effects and regulating aquaporin-4 membrane distribution. Targeting MMP-9 may represent a promising strategy to prevent postoperative cognitive dysfunction by integrating multiple protective mechanisms.


Ethylene and Abscisic Acid Signaling Pathways Differentially Influence Tomato Resistance to Combined Powdery Mildew and Salt Stress.
There is currently limited knowledge on the role of hormones in plants responses to combinations of abiotic and pathogen stress factors. This study focused on the response of tomato near-isogenic lines (NILs) that carry the Ol-1, ol-2, and Ol-4 loci, conferring resistance to tomato powdery mildew (PM) caused by Oidium neolycopersici, to combined PM and salt stress. These NILs were crossed with the notabilis (ABA-deficient), defenceless1 (JA-deficient), and epinastic (ET overproducer) tomato mutants to investigate possible roles of hormone signaling in response to combined stresses. In the NILs, marker genes for hormonal pathways showed differential expression patterns upon PM infection. The epinastic mutation resulted in breakdown of resistance in NIL-Ol-1 and NIL-ol-2. This was accompanied by reduced callose deposition, and was more pronounced under combined salt stress. The notabilis mutation resulted in H2O2 overproduction and reduced susceptibility to PM in NIL-Ol-1 under combined stress, but lead to higher plant growth reduction under salinity and combined stress. Resistance in NIL-ol-2 was compromised by the notabilis mutation, which was potentially caused by reduction of callose deposition. Under combined stress the compromised resistance in NIL-ol-2 was restored. PM resistance in NIL-Ol-4 remained robust across all mutant and treatment combinations. Hormone signaling is critical to the response to combined stress and PM, in terms of resistance and plant fitness. ABA appears to be at the crossroads of disease susceptibility/senescence and plant performance under combined stress These gained insights can aid in narrowing down targets for improving crop performance under stress combinations.


Stability of early identified aggressive victim status in elementary school and associations with later mental health problems and functional impairments.
Aggressive victims-children who are both perpetrators and victims of peer aggression-experience greater concurrent mental health problems and impairments than children who are only aggressive or only victimized. The stability of early identified aggressive victim status has not been evaluated due to the fact that most studies of aggressor/victim subgroups have focused on preadolescents and/or adolescents. Further, whether children who exhibit early and persistent patterns of aggression and victimization continue to experience greater mental health problems and functional impairments through the transition to adolescence is not known. This study followed 344 children (180 girls) previously identified as socially adjusted, victims, aggressors, or aggressive victims at Grade 1 (Burk et al. 2008) to investigate their involvement in peer bullying through Grade 5. The children, their mothers, and teachers reported on children's involvement in peer aggression and victimization at Grades 1, 3, and 5; and reported on internalizing symptoms, externalizing symptoms, inattention and impulsivity, as well as academic functioning, physical health, and service use at Grades 5, 7, and 9. Most children categorized as aggressive victims in Grade 1 continued to be significantly involved in peer bullying across elementary school. Children with recurrent aggressive victim status exhibited higher levels of some mental health problems and greater school impairments across the adolescent transition when compared to other longitudinal peer status groups. This study suggests screening for aggressive victim status at Grade 1 is potentially beneficial. Further early interventions may need to be carefully tailored to prevent and/or attenuate later psychological, academic, and physical health problems.


Fluorine- and rhenium-containing geldanamycin derivatives as leads for the development of molecular probes for imaging Hsp90.
Heat shock protein 90 (Hsp90) is an ATP-dependent molecular chaperone responsible for protein quality control in cells. Hsp90 has been shown to be overexpressed in many human cancers. This has prompted extensive research on Hsp90 inhibitors as novel anticancer agents and, more recently, the development of molecular probes for imaging Hsp90 expression in vivo. This work describes the development of various fluorine-containing and rhenium-containing geldanamycin derivatives as leads for the development of corresponding (18)F-labeled and (99m)Tc-labeled PET and SPECT probes for molecular imaging of Hsp90 expression. All compounds were evaluated in an in vitro ATPase activity assay using Hsp90 isoform Hsp82p. Fluorobenzoylated geldanamycin derivative 5 displayed comparable inhibitory potency like parent compound geldanamycin.


Red Cell Alloantibody Screening: Comparative Analysis of Three Different Technologies.
The detection of irregular antibody is a critical issue in the management of red blood cell transfusion according to the Type & Screen (T&S) practice. In order to implement the T&S procedure at our blood bank, we compared three different automated analyzers based on column agglutination technique (CAT) or solid phase red cell adherence assay (SPACA) methods. Pre-transfusion antibody screening was performed in 986 patients candidate to elective surgery at low risk for red blood cell transfusion. We tested the following kits: the three-cell panel micro-CAT system ID-DiaCell I-II-III (DiaMed), the four-cell panel solid-phase system Capture-R Ready Screen-4 (Immucor), and the four-cell panel micro-CAT system Serascan Diana-4 (Grifols). Positive results were further investigated using corresponding identification panels, and discrepant results were investigated with all the antibody identification systems. Among 986 samples, we observed 967 concordant negative results (98.1%), 8 concordant positive results (0.8% of cases), and 11 discrepant results (1.1%). Among discrepant samples, an alloantibody could been identified in two patents (anti-M, detected by Serascan Diana-4 and ID-DiaCell I, II, III; anti-Kpa, detected by Capture-R Ready Screen-4 and Serascan Diana-4). Among the evaluated technologies, the four-cell panel micro-CAT system displayed the highest sensitivity and specificity with an optimal negative predictive value. These features might be relevant to the routine implementation of the T&S transfusion strategy.


Theoretical application of z-dependent gain coefficient to describe amplified spontaneous emission.
Based on the geometrical modeling of the unified gain coefficient and the reported amplified spontaneous emission (ASE) output energy measurement ε(ASE) versus amplifying excitation length, l(AMP) in a KrF laser oscillator, we managed, as an example, to explain the ASE output energy behavior both numerically and analytically. In this approach, introducing the ASE gain-coefficient profile for the KrF laser, g(0,KrF)(ASE), was not avoidable. It was found that while the g(0,KrF)(ASE) profile follows the introduced gain-modeling formulation, it is, however, slightly lower than the KrF laser gain profile, g(0,KrF)(exp), deduced from the measurements reported by different researchers. The present approach, up to the present time, is able to explain all of the existing ambiguities on understanding the ASE behavior.


Individual and network characteristic associated with hospital-acquired Middle East Respiratory Syndrome coronavirus.
During outbreaks of infectious diseases, transmission of the pathogen can form networks of infected individuals connected either directly or indirectly. Network centrality metrics were used to characterize hospital-acquired Middle East Respiratory Syndrome Coronavirus (HA-MERS) outbreaks in the Kingdom of Saudi Arabia between 2012 and 2016. Covariate-adjusted multivariable logistic regression models were applied to assess the effect of individual level risk factors and network level metrics associated with increase in length of hospital stay and risk of deaths from MERS. About 27% of MERS cases were hospital acquired during the study period. The median age of healthcare workers and hospitalized patients were 35 years and 63 years, respectively, Although HA-MERS were more connected, we found no significant difference in degree centrality metrics between HA-MERS and non-HA-MERS cases. Pre-existing medical conditions (adjusted Odds ratio (aOR)=2.43, 95% confidence interval: (CI) [1.11-5.33]) and hospitalized patients (aOR=29.99, 95% CI [1.80-48.65]) were the strongest risk predictors of death from MERS. The risk of death associated with 1-day increased length of stay was significantly higher for patients with comorbidities. Our investigation also revealed that patients with an HA-MERS infection experienced a significantly longer hospital stay and were more likely to die from the disease. Healthcare worker should be reminded of their potential role as hubs for pathogens because of their proximity to and regular interaction with infected patients. On the other hand, this study has shown that while healthcare workers acted as epidemic attenuators, hospitalized patients played the role of an epidemic amplifier.


Identification of replication-competent strains of simian immunodeficiency virus lacking multiple attachment sites for N-linked carbohydrates in variable regions 1 and 2 of the surface envelope protein.
Carbohydrates comprise about 50% of the mass of gp120, the external envelope glycoprotein of simian immunodeficiency virus (SIV) and human immunodeficiency virus. We identified 11 replication-competent derivatives of SIVmac239 lacking two, three, four, or five potential sites for N-linked glycosylation. These sites were located within and around variable regions 1 and 2 of the surface envelope protein of the virus. Asn (AAT) of the canonical N-linked glycosylation recognition sequence (Asn X Ser/Thr) was changed in each case to the structurally similar Gln (CAG or CAA) such that two nucleotide changes in the codon would be required for reversion. Replication of one triple mutant (g456), however, was severely impaired. A revertant of the g456 mutant was recovered from CEMx174 cells with a Met-to-Val compensatory substitution at position 144, 2 amino acids upstream of attachment site 5. Thus, a debilitating loss of sites for N-linked glycosylation can be compensated for by amino acid changes not involving the Asn-X-Ser/Thr consensus motif. These results provide a framework to begin testing the hypothesis that carbohydrates form a barrier that can limit the humoral immune responses to the virus.


Early results of fenestrated endovascular repair of juxtarenal aortic aneurysms in the United Kingdom.
Fenestrated endovascular repair of abdominal aortic aneurysms has been proposed as an alternative to open surgery for juxtarenal and pararenal abdominal aortic aneurysms. At present, the evidence base for this procedure is predominantly limited to single-center or single-operator series. The aim of this study was to present nationwide early results of fenestrated endovascular repair in the United Kingdom. All patients who underwent fenestrated endovascular repair between January 2007 and December 2010 at experienced institutions in the United Kingdom(>10 procedures) were retrospectively studied by use of the GLOBALSTAR database. Site-reported data relating to patient demographics, aneurysm morphology, procedural details, and outcome were recorded. Data from 318 patients were obtained from 14 centers. Primary procedural success was achieved in 99% (316/318); perioperative mortality was 4.1%, and intraoperative target vessel loss was observed in 5 of 889 target vessels (0.6%). The early reintervention (<30 days) rate was 7% (22/318). There were 11 deaths during follow-up; none were aneurysm-related. Survival by Kaplan-Meier analysis was 94% (SE 0.01), 91% (0.02), and 89% (0.02) at 1, 2, and 3 years, respectively. Freedom from target vessel loss was 93% (0.02), 91% (0.02), and 85% (0.06), and freedom from late secondary intervention (>30 days) was 90% (0.02), 86% (0.03), and 70% (0.08) at 1, 2, and 3 years. In this national sample, fenestrated endovascular repair has been performed with a high degree of technical and clinical success. Late survival and target vessel patency are satisfactory. These results support continued use and evaluation of this technique for juxtarenal aneurysms, but illustrate the need for a more robust evidence base.


Hypothalamic glial-to-neuronal signaling during puberty: influence of alcohol.
Mammalian puberty requires complex interactions between glial and neuronal regulatory systems within the hypothalamus that results in the timely increase in the secretion of luteinizing hormone releasing hormone (LHRH). Assessing the molecules required for the development of coordinated communication networks between glia and LHRH neuron terminals in the basal hypothalamus, as well as identifying substances capable of affecting cell-cell communication are important. One such pathway involves growth factors of the epidermal growth factor (EGF) family that bind to specific erbB receptors. Activation of this receptor results in the release of prostaglandin-E(2) (PGE(2)) from adjacent glial cells, which then acts on the nearby LHRH nerve terminals to elicit release of the peptide. Another pathway involves novel genes which synthesize adhesion/signaling proteins responsible for the structural integrity of bi-directional glial-neuronal communication. In this review, we will discuss the influence of these glial-neuronal communication pathways on the prepubertal LHRH secretory system, and furthermore, discuss the actions and interactions of alcohol on these two signaling processes.


Isolation and characterization of a native cell wall complex from Neisseria meningitidis.
A cell wall complex has been isolated by gentle methods from both the medium supernatant fluid and whole organisms of Neissieria meningitidis cultures. The two types of preparations have been shown to be essentially identical on the basis of chemical composition, electron microscopy, and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE). Four major components were identified in the complex: group-specific polysaccharide (4 to 10%), protein (45 to 65%), lipopolysaccharide (10 to 25%), and lipid (15 to 30%). The whole complex was found to be immunogenic in rabbits and to elicit production of antibody directed against the protein, the group-specific polysaccharide, and the lipopolysaccharide components. The isolated protein component was also found to be immunogenic in rabbits and to elicit production of serotype-specific antibody. The protein component was found to produce a band pattern in SDS-PAGE that is simple, reproducible, and strain dependent. The lipopolysaccharide component was found to have chemical and biological properties characteristic of bacterial endotoxin. We propose that this complex is representative of the outer trilaminar membrane of the meningococcal cell envelope in its native state.


Are medical students equipped for digital studies?Have their hopes and fears been confirmed during Covid-19? What should we consider in the future?
Digitisation in the education of future doctors was still in its infancy before the Covid pandemic. For the successful implementation of digital teaching, students need the technical equipment and the necessary skills to use it in a meaningful way. Furthermore, it requires a willingness to adapt the learning environment and to take responsibility for self-directed learning. At the beginning of 2020, faculties were forced to convert all teaching to digital formats. Initial research shows that students prefer face-to-face teaching. To determine whether medical students were prepared for digital studies and what should be considered for the future, we analysed surveys at the beginning of online studies and after two Corona semesters at a medical faculty. We were able to show that although our students had good technology equipment, they had a rather negative attitude towards online teaching for various reasons and developed negative emotions. Deficits in design of educational material, and personal learning habits raised concern. A lack of guidance and a lack of interaction with fellow students contributed to this. Adjustments in these areas will be necessary in the future to provide students with positive access to digital studies and thus increase learning success.


[Morphofunctional development of the hippocampal formation of newborn mice in organotypical tissue culture].
The cytoarchitectonic structures, fiber pathways, spontaneous and evoked neuronal activity in the organotypic explants of the newborn mouse hippocampus were studied. After 2-3 weeks in vitro it was possible to see in the explants main fascicles (alvear tract, perforant path, axons of granular cells) and cytoarchitectonic formations (pyramidal cell layer of hippocampus, infra- und suprapyramidal limbs of dentate gyrus granular cells). Electrical stimuli applied to the entorhinal cortex and fascia dentata evoked short- and long-latency responses in hippocampal neurons beginning from the 5-6th day in vitro. The short-latency responses were presumably monosynaptic, the long-lasting ones, polysnaptic. The latter were usually changed by repetitive stimulation. These data suggest that appearance of evoked activity during cultivation reflects the formation in vitro of the perforant path and mossy fibers and the maturation of the synaptic contacts. Thus, organotypic explants of the hippocampus represent a useful model system for morphological and electrophysiological studies in systems of interneuronal connections formed de novo as a result of autonomical postnatal development in vitro.


SAR versus S(inc): What is the appropriate RF exposure metric in the range 1-10 GHz? Part I: Using planar body models.
This is the first of two articles addressing the most appropriate crossover frequency at which incident power flux density (S(inc)) replaces the spatial peak value of the specific energy absorption rate (SAR) averaged over 1 or 10 g (i.e., peak 1 or 10 g SAR) as the basic restriction for protecting against radiofrequency (RF) heating effects in the 1-10 GHz range. Our general approach has been to compare the degree of correlation between these basic restrictions and the peak induced tissue temperature rise (DeltaT) for a representative range of population/exposure scenarios. In this article we particularly address the effect of human population diversity in the thickness of body tissue layers at eight different sites of the body. We used a Monte Carlo approach to specify 32000 models (400 models for each of 8 body sites for 10 frequencies) which were representative of tissue thicknesses for age (18-74 years) and sex at the eight body sites. Histogram distributions of S(inc) and peak 1 and 10 g SAR corresponding to a peak 1 degrees C temperature rise were obtained from RF and thermal analyses of 1D multiplanar models exposed to a normally incident plane wave ranging from 1 to 10 GHz in thermo-neutral environmental conditions. Examination of the distribution spread of the histograms indicated that peak SAR was a better predictor of peak tissue temperature rise across the entire 1-10 GHz frequency range than S(inc), as indicated by the smaller spread in its histogram distributions, and that peak 10 g SAR was a slightly better predictor than peak 1 g SAR. However, this result must be weighed against partly conflicting indications from complex body modeling in the second article of this series, which incorporates near-field effects and the influence of complex body geometries.


Observation of magnetoreceptive behavior in a multicellular magnetotactic prokaryote in higher than geomagnetic fields.
The magnetotactic multicellular prokaryote (MMP), a motile aggregate of bacterial cells, is known to exhibit an unusual "ping-pong" motility in magnetic fields greater than the earth's field. This motility is characterized by rapid excursions, opposite the direction of an applied magnetic field, and slower returns along the direction of the magnetic field. We have carried out detailed observations of the time and spatial dependence of the ping-pong motility and find 1), the outward and return excursions exhibit a uniform deceleration and acceleration, respectively; 2), the probability per unit time of an MMP undergoing a ping-pong excursion increases monotonically with the field strength; and 3), the outward excursions exhibit a very unusual distance distribution which is dependent on the magnetic field strength. At any given field strength, a characteristic distance is observed, below which very few excursions occur. Beyond this distance, there is a rapid increase in the number of excursions with an exponentially decaying distribution. These observations cannot be explained by conventional magnetotaxis, i.e., a physical directing torque on the organism, and suggest a magnetoreceptive capability of the MMP.


Swing Velocity Profiles of Small Limbs Can Arise from Transient Passive Torques of the Antagonist Muscle Alone.
In large limbs, changing motor neuron activity typically controls within-movement velocity. For example, sequential agonist-antagonist-agonist motor neuron firing typically underlies the slowing often present at the end of human reaches. In physiological movements of large limbs, antagonistic muscle passive torque is generally negligible. In small limbs, alternatively, passive torques can determine limb rest position, generate restoring movements to it, and decrease agonist-generated movement amplitude and velocity maxima. These observations suggest that, in small limbs, passive forces might also control velocity changes within movements. We investigated this issue in stick insect middle leg femur-tibia (FT) joint. During swing, the FT joint extensor muscle actively shortens and the flexor muscle passively lengthens. As in human reaching, after its initial acceleration, FT joint velocity continuously decreases. We measured flexor passive forces during imposed stretches spanning the ranges of FT joint angles, angular velocities, and movement amplitudes present in leg swings. The viscoelastic "transient" passive force that occurs during and soon after stretch depended on all three variables and could be tens of times larger than the "steady-state" passive force commonly measured long after stretch end. We combined these data, the flexor and extensor moment arms, and an existing extensor model to simulate FT joint swing. To measure only passive (flexor) muscle-dependent effects, we used constant extensor activations in these simulations. In simulations using data from ten flexor muscles, flexor passive torque could always produce swings with, after swing initiation, continuously decreasing velocities. Antagonist muscle passive torques alone can thus control within-movement velocity.


Serum Per- and Polyfluoroalkyl Substances Are Associated with Increased Hearing Impairment: A Re-Analysis of the National Health and Nutrition Examination Survey Data.
Although studies have shown that per- and polyfluoroalkyl substances (PFAS) are potential environmental ototoxicants, epidemiologic study has been limited. I conducted a cross-sectional study to re-examine the associations between PFAS and hearing impairment. Data were obtained from the National Health and Nutrition Examination Survey (NHANES) 1999-2000, 2003-06, 2009-12, and 2015-16. Perfluorooctanoic acid (PFOA), perfluorooctane sulfonic acid (PFOS), perfluorohexane sulfonic acid (PFHxS), and perfluorononanoic acid (PFNA) were measured in serum samples. Participants were divided into quartiles for each PFAS. Air conduction pure-tone audiometry was administered. Hearing impairment (1: yes, 0: no) was defined as a hearing threshold of more than 25 dB at 500, 1000, 2000, 4000, and 8000 Hz in the worse ear. I assessed the relation of serum PFAS with hearing impairment by the generalized linear mixed model with a logit link and binary distribution. Tests for linear trend across quartiles of serum PFAS were conducted using the median serum PFAS in each quartile as a continuous variable. After adjusting for age, sex, body mass index, education, ethnicity group, and family income, I found positive correlations between PFOA and hearing impairment at 2000 Hz (p-trend < 0.01) and 3000 Hz (p-trend = 0.02); between PFOS and hearing impairment at 500 Hz (p-trend < 0.01), 2000 Hz (p-trend < 0.0001) and 3000 Hz (p-trend = 0.02); between PFNA and hearing impairment at 2000 Hz (p-trend = 0.05), 3000 Hz (p-trend < 0.01), 4000 Hz (p-trend = 0.02), and 8000 Hz (p-trend < 0.01); between PFHxS and hearing impairment at 500 Hz (p-trend = 0.04), 1000 Hz (p-trend = 0.03), and 2000 Hz (p-trend < 0.01). However, some of the findings were not significant when only comparing the highest with the lowest quartile of PFASs. In conclusion, several background serum PFASs are positively correlated with hearing impairment in the United States adult population.


The impact of intraoperative N-acetylcysteine on opioid consumption following spine surgery: a randomized pilot trial.
Aim: N-acetylcysteine (NAC) decreases inflammation and could augment perioperative analgesia. Materials & methods: This prospective pilot trial examined postoperative opioid consumption at 12 h following intraoperative NAC. In phase I, 20 adults scheduled for posterior spine surgery were randomized to NAC (0, 50, 100 and 150 mg/kg) to determine the optimal dose. In phase II, 30 patients were randomized to placebo or NAC (150 mg/kg). Opioid consumption, pain ratings and time to opioid rescue were recorded. Results: Postoperative opioid consumption was reduced in the NAC group 19.3% at 12 h and 20% at 18 and 36 h. Opioid consumption was reduced 22-24% in the NAC group at all times after adjusting for intraoperative opioid administration. NAC subjects reported lower pain scores relative to placebo. Conclusion: Subjects randomized to NAC consumed less postoperative opioids and reported less pain versus placebo. Larger randomized controlled trials are needed to further evaluate NAC for analgesia. Clinical Trial Registration: NCT04562597 (ClinicalTrials.gov).


A best-worst scaling experiment to prioritize caregiver concerns about ADHD medication for children.
The objective of this feasibility study was to develop and pilot an instrument to elicit caregivers' priorities when initiating attention-deficit hyperactivity disorder (ADHD) medication for their child. A best-worst scaling experiment was used to rank competing priorities when initiating ADHD medicine. Forty-six participants were recruited for a two-phase study involving survey development (phase 1, N=21) and the survey pilot (phase 2, N=25). Best-worst scores and 95% confidence intervals indicating the relative importance of 16 concerns were determined, and t tests were used to determine the scores' significance. The significance of best-worst scores for most concerns indicated that the choices were purposeful. Concerns about helping the child become a successful adult, having a doctor who addresses caregivers' concerns, and improving school behavior were ranked highest. The best-worst scaling method can elicit priorities for children's mental health treatment. Future work using this method will guide family-centered care.


G alpha(q)-coupled receptors in human atrium function through protein kinase C epsilon and delta.
Cardiac G alpha(q)-coupled receptors (such as endothelin, angiotensin, and alpha1-adrenergic receptors) mediate cardiac inotropy and chronotropy, as well as the development of hypertrophy. These receptors signal through protein kinase C (PKC), a family of 12 isozymes including PKC alpha, beta I, beta II, gamma, delta, epsilon, theta, eta, lambda, iota, zeta, and mu. Of these PKC isozymes, alpha, beta II, gamma, epsilon, delta, and zeta have been implicated in signaling through cardiac G alpha(q)-coupled receptors in various animal models. However, the profile of which isozymes are activated by a given G alpha(q)-coupled receptor varies among animal species. Thus, these results can not be extrapolated to human heart. In this study, we examine PKC isozymes activated by three different G alpha(q)-coupled receptors in human atrial tissue. Live atrial appendages obtained from the operating room were sliced and treated with agonists of G alpha(q)-coupled receptors, and cellular redistribution of PKC isozymes was examined by immunoblotting. We find that stimulation of G alpha(q)-coupled receptors in human atrium activates PKC epsilon and delta only, under both acute (5 min) and longer (35 min) stimulations. Further, PKC epsilon and delta exhibit distinct subcellular redistribution patterns; while both translocate to the plasma membrane upon G alpha(q) stimulation, PKC delta also redistributes to mitochondria. We conclude that PKC epsilon and delta are the main PKC isozymes involved in G alpha(q)-mediated signaling in human atria.


Computational methods for constructing protein structure models from 3D electron microscopy maps.
Protein structure determination by cryo-electron microscopy (EM) has made significant progress in the past decades. Resolutions of EM maps have been improving as evidenced by recently reported structures that are solved at high resolutions close to 3Å. Computational methods play a key role in interpreting EM data. Among many computational procedures applied to an EM map to obtain protein structure information, in this article we focus on reviewing computational methods that model protein three-dimensional (3D) structures from a 3D EM density map that is constructed from two-dimensional (2D) maps. The computational methods we discuss range from de novo methods, which identify structural elements in an EM map, to structure fitting methods, where known high resolution structures are fit into a low-resolution EM map. A list of available computational tools is also provided.


Who is paid in pay-for-performance? Inequalities in the distribution of financial bonuses amongst health centres in Zimbabwe.
Although pay-for-performance (P4P) schemes have been implemented across low- and middle-income countries (LMICs), little is known about their distributional consequences. A key concern is that financial bonuses are primarily captured by providers who are already better able to perform (for example, those in wealthier areas), P4P could exacerbate existing inequalities within the health system. We examine inequalities in the distribution of pay-outs in Zimbabwe's national P4P scheme (2014-2016) using quantitative data on bonus payments and facility characteristics and findings from a thematic policy review and 28 semi-structured interviews with stakeholders at all system levels. We found that in Zimbabwe, facilities with better baseline access to guidelines, more staff, higher consultation volumes and wealthier and less remote target populations earned significantly higher P4P bonuses throughout the programme. For instance, facilities that were 1 SD above the mean in terms of access to guidelines, earned 90 USD more per quarter than those that were 1 SD below the mean. Differences in bonus pay-outs for facilities that were 1 SD above and below the mean in terms of the number of staff and consultation volumes are even more pronounced at 348 USD and 445 USD per quarter. Similarly, facilities with villages in the poorest wealth quintile in their vicinity earned less than all others-and 752 USD less per quarter than those serving villages in the richest quintile. Qualitative data confirm these findings. Respondents identified facility baseline structural quality, leadership, catchment population size and remoteness as affecting performance in the scheme. Unequal distribution of P4P pay-outs was identified as having negative consequences on staff retention, absenteeism and motivation. Based on our findings and previous work, we provide some guidance to policymakers on how to design more equitable P4P schemes.


[External canthopexy using the Edgerton-Montandon procedure in lagophthalmos of leprosy patients. Technique and indications. Apropos of 30 cases].
This paper deals with the results observed in 21 ancient leprosy patients suffering from lagophthalmos (13 of whom suffered from bilateral lagophthalmos) and treated by the Edgerton-Montandon surgical procedure which associates lateral canthopexy and tarsorraphy. Eighteen of the 21 treated patients were reviewed at one month after the procedure and, overall, results could be evaluated for 30 eyes. Improvement was noted in all of the 30 eyes and, globally, the residual palpebral fissure (during voluntary closing of the eyes by the patient) decreased from 6.7 mm before the procedure to 1.8 after the procedure. The following recommendations may be proposed. For young patients with intact corneal sensation, the Gillies procedure remains the procedure of choice to correct lagophthalmos. For older patients with corneal anesthesia, at high risk of blindness, the Edgerton-Montandon procedure should be recommended.


In search for new bonding modes of the methylenedithiolato ligand: novel tri- and tetra-metallic clusters.
Building upon our earlier results on the chemistry of diruthenium analogue of pentaborane (9) with heterocumulenes, we continued to investigate the reactivity of arachno-[(Cp*Ru)2(B3H8)(CS2H)], 1, (Cp* = η(5)-C5Me5) towards group 7 and 8 transition metal carbonyl compounds under photolytic and thermolytic conditions. The metal carbonyl compounds show diverse reactivity pattern with arachno-1. For example, the photolysis of arachno-1 with [Re2(CO)10] yielded [(Cp*Ru)2B3H5(CH2S2){Re(CO)4}2], 2, [(Cp*RuCO)2(μ-H)2(CH2S2){Re(CO)4}{Re(CO)3}], 3 and [(Cp*Ru)2(μ-CO)(μ-H)(CH2S2){Re(CO)3}], 4. The geometry of 2 with a nearly planar eight-membered ring containing heavier transition metals rhenium, ruthenium is unprecedented. Compounds 3 and 4 can be considered as M4-quadrilateral and M3-triangle with a methylenedithiolato ligand attached to the metal centres, respectively. [Mn2(CO)10], on the other hand, reacts with arachno-1 to yield heterometallic binuclear [(Cp*RuCO){Mn(CO)4}(μ-H)(SCH3)], 5 and homocubane [(Cp*Ru)2{Mn(CO)3}-(CS2H2)B3H4], 6. In an attempt to generate group 8 analogues of 2-5, we performed the reaction of arachno-1 with [Fe2(CO)9] and [Ru3(CO)12]. Although, the objective of isolating analogous compounds was not achieved, the reaction with [Fe2(CO)9] led to novel tetrahedral cluster [(Cp*RuCO){(Fe(CO)3}2S(μ-H)], 7. [Ru3(CO)12], in contrast, yielded known compounds [{Cp*Ru(CO)}2B2H6], 9 and [Cp*Ru(CO)2]2, 10. All the cluster compounds have been characterized by mass spectrometry, IR, and (1)H, (11)B, and (13)C NMR spectroscopy, and the geometric structures were unequivocally established by crystallographic analysis of 2-5 and 7.