A comparison of mass spectrometry based hydrogen deuterium exchange methods for probing the cyclophilin A cyclosporin complex.
Direct infusion electrospray ionisation mass spectrometry (DI-ESI-MS) techniques provide an increasingly popular route to determine quantitative information on protein-protein and protein-ligand interactions. When combined with hydrogen deuterium exchange (HDX), details on protein stability and complex conformation can be obtained; however, complexes retained by ESI-MS are not always representative of those in solution and care must be taken in interpreting gas phase results. Zhu et al. [1] and Powell and Fitzgerald [2] have outlined LC-MS based techniques to probe the solution phase properties of the protein-ligand system in question. We here have taken the well characterised soluble immunophilin protein cyclophilin A, and examined it in complex with its endogenous ligand cyclosporin A. This ligand is widely used as an immunosuppressant following organ transplant, and the complex provides a basis for drug discovery efforts. We have used direct infusion, coupled with HDX, gas phase HDX and also the LC-HDX techniques PLIMSTEX and SUPREX. Results from each of these four HDX methodologies are presented here and discussed critically. From our direct infusion we find that there are 2 observable hydrogen populations in the protein, a very fast exchanging population, and a slower group. The exchange rate of both is lowered in the presence of the ligand. For PLIMSTEX we find a K(d) for ligand binding of 321 ± 128 nM, which is within one order of magnitude of values previously reported. SUPREX under a variety of conditions provides a range of K(d) values, but when we average these for experimental error we obtain a K(d) of 7.11 ± 0.29 nM which agrees well with measurements from other studies including via SUPREX. Finally gas phase HDX of the native complex shows more than 3 distinct populations of exchangeable hydrogens, for both the apo- and the holo protein consistent with an unfolding and refolding of the protein in the gas phase. The different techniques are compared with respect to the advantages and disadvantages they bring to the study of this protein-ligand system.


Mutagenesis of the dengue virus type 2 NS5 methyltransferase domain.
The Flavivirus NS5 protein possesses both (guanine-N7)-methyltransferase and nucleoside-2'-O methyltransferase activities required for sequential methylation of the cap structure present at the 5' end of the Flavivirus RNA genome. Seventeen mutations were introduced into the dengue virus type 2 NS5 methyltransferase domain, targeting amino acids either predicted to be directly involved in S-adenosyl-l-methionine binding or important for NS5 conformation and/or charged interactions. The effects of the mutations on (i) (guanine-N7)-methyltransferase and nucleoside-2'-O methyltransferase activities using biochemical assays based on a bacterially expressed NS5 methyltransferase domain and (ii) viral replication using a dengue virus type 2 infectious cDNA clone were examined. Clustered mutations targeting the S-adenosyl-l-methionine binding pocket or an active site residue abolished both methyltransferase activities and viral replication, demonstrating that both methyltransferase activities utilize a single S-adenosyl-l-methionine binding pocket. Substitutions to single amino acids binding S-adenosyl-l-methionine decreased both methyltransferase activities by varying amounts. However, viruses that replicated at wild type levels could be recovered with mutations that reduced both activities by >75%, suggesting that only a threshold level of methyltransferase activity was required for virus replication in vivo. Mutation of residues outside of regions directly involved in S-adenosyl-l-methionine binding or catalysis also affected methyltransferase activity and virus replication. The recovery of viruses containing compensatory second site mutations in the NS5 and NS3 proteins identified regions of the methyltransferase domain important for overall stability of the protein or likely to play a role in virus replication distinct from that of cap methylation.


The potential of metatranscriptomics for identifying screening targets for bacterial vaginosis.
The ribosomal RNA content of a sample collected from a woman with bacterial vaginosis (BV) was analysed to determine the active microbial community, and to identify potential targets for further screening. The sample from the BV patient underwent total RNA extraction, followed by physical subtraction of human rRNA and whole transcriptome amplification. The metatranscriptome was sequenced using Roche 454 titanium chemistry. The bioinformatics pipeline MG-RAST and desktop DNA analysis platforms were utilised to analyse results. Bacteria of the genus Prevotella (predominately P. amnii) constituted 36% of the 16S rRNA reads, followed by Megasphaera (19%), Leptotrichia/Sneathia (8%) and Fusobacterium (8%). Comparison of the abundances of several bacteria to quantitative PCR (qPCR) screening of extracted DNA revealed comparable relative abundances. This suggests a correlation between what was present and transcriptionally active in this sample: however distinct differences were seen when compared to the microbiome determined by 16S rRNA gene amplicon sequencing. To assess the presence of P. amnii in a larger pool of samples, 90 sexually active women were screened using qPCR. This bacterium was found to be strongly associated with BV (P<0.001, OR 23.3 (95%CI:2.9-190.7)) among the 90 women. This study highlighted the potential of metatranscriptomics as a tool for characterising metabolically active microbiota and identifying targets for further screening. Prevotella amnii was chosen as an example target, being the most metabolically active species present in the single patient with BV, and was found to be detected at a high concentration by qPCR in 31% of cohort with BV, with an association with both oral and penile-vaginal sex.


Titanium carbide nanocrystals in circumstellar environments.
Meteorites contain micrometer-sized graphite grains with embedded titanium carbide grains. Although isotopic analysis identifies asymptotic giant branch stars as the birth sites of these grains, there is no direct observational identification of these grains in astronomical sources. We report that infrared wavelength spectra of gas-phase titanium carbide nanocrystals derived in the laboratory show a prominent feature at a wavelength of 20.1 micrometers, which compares well to a similar feature in observed spectra of postasymptotic giant branch stars. It is concluded that titanium carbide forms during a short (approximately 100 years) phase of catastrophic mass loss (>0.001 solar masses per year) in dying, low-mass stars.


Characterization of Physicians That Might Be Reluctant to Propose HIV Cure-Related Clinical Trials with Treatment Interruption to Their Patients? The ANRS-APSEC Study.
HIV cure-related clinical trials (HCRCT) with analytical antiretroviral treatment interruptions (ATIs) have become unavoidable. However, the limited benefits for participants and the risk of HIV transmission during ATI might negatively impact physicians' motivations to propose HCRCT to patients. Between October 2016 and March 2017, 164 French HIV physicians were asked about their level of agreement with four viewpoints regarding HCRCT. A reluctance score was derived from their answers and factors associated with reluctance identified. Results showed the highest reluctance to propose HCRCT was among physicians with a less research-orientated professional activity, those not informing themselves about cure trials through scientific literature, and those who participated in trials because their department head asked them. Physicians' perceptions of the impact of HIV on their patients' lives were also associated with their motivation to propose HCRCT: those who considered that living with HIV means living with a secret were more motivated, while those worrying about the negative impact on person living with HIV's professional lives were more reluctant. Our study highlighted the need to design a HCRCT that minimizes constraints for participants and for continuous training programs to help physicians keep up-to-date with recent advances in HIV cure research.


Gem GTPase and tau: morphological changes induced by gem GTPase in cho cells are antagonized by tau.
A series of observations have indicated that tau, one of the major microtubule-associated proteins, is involved in neuronal cell morphogenesis and axonal maintenance. Tau is also the major component of paired helical filaments found in brains affected by Alzheimer's disease. To explore an as yet unidentified role of tau in vivo, approximately 11,000 mRNAs were profiled from tau-deficient mouse brains and compared with those from control brains at the same ages. The expression of Gem GTPase, a small GTP-binding protein of the ras superfamily, was significantly increased in the brains of tau-deficient mice at 8 weeks of age. Because Gem GTPase is a negative regulator of the Rho-Rho kinase pathway for cytoskeletal organization, this protein was transiently overexpressed in Chinese hamster ovary cells that do not express tau. Overexpression of Gem GTPase induced a marked elongation of Chinese hamster ovary cells, and simultaneous expression of tau eliminated this effect, although tau did not bind directly to Gem GTPase. This anti-elongation activity of tau was attributed to its microtubule-binding domain, and homologous domains of microtubule-associated proteins 2 and 4 exhibited similar antagonistic activities. Taken together, the present results indicate that the level of Gem GTPase and its cell elongation activity are modulated by tau and suggest that tau may be involved in a Gem GTPase-mediated signal transduction pathway.


Leaving "Drug Abuse" behind: A Theoretical and Methodological Heuristic to Selecting "Problem Drug Use" or "Drug Misuse" as Alternative Terms.
Background: Historically, drug abuse described the negative consequences of drug use as a proxy measure of maladaptive behavior that manifests within the addiction process. Negative consequences can span multiple domains including physical (e.g., blacking out), psychological (e.g., mood shifts), interpersonal (e.g., relationship problems), social (e.g., drinking and driving), and economic (e.g., job loss) problems. Objective: In determining the appropriate terminology to describe the phenomenon under investigation, researchers should consider (1) the theoretical objective (i.e., substance use construct under investigation) and (2) the research methodology being employed. Results: Problem substance use is appropriate for describing the outcome of interest when conducting cross-sectional, or pre-post, designs where investigators are interested in a snapshot of problems associated with substance use behavior. Conversely, substance misuse reflects a pattern of negative consequences over time and is appropriate when conducting longitudinal research with three or more time points. Here, substance misuse captures a pattern of substance use problems over time that may be indicative of a substance use disorder. These terms should be distinguished from studies investigating the frequency (how often) and quantity (how much) of a substance is being used where risky substance use is appropriate for cross-sectional studies and heavy substance use is appropriate for longitudinal studies. Conclusion: This framework is intended to describe the phenomena being investigated (i.e., the variable) and not the person or people experiencing a substance use problem. We should continue the dialogue of semantics in science realizing that our choice of words has important "real world" ramifications for the populations we serve.


Study protocol: Cost effectiveness of two strategies to implement the NVOG guidelines on hypertension in pregnancy: An innovative strategy including a computerised decision support system compared to a common strategy of professional audit and feedback, a randomized controlled trial.
Hypertensive disease in pregnancy remains the leading cause of maternal mortality in the Netherlands. Seventeen percent of the clinical pregnancies are complicated by hypertension and 2% by preeclampsia. The Dutch Society of Obstetrics and Gynaecology (NVOG) has developed evidence-based guidelines on the management of hypertension in pregnancy and chronic hypertension. Previous studies showed a low adherence rate to other NVOG guidelines and a large variation in usual care in the different hospitals. An explanation is that the NVOG has no general strategy of practical implementation and evaluation of its guidelines. The development of an effective and cost effective implementation strategy to improve adherence to the guidelines on hypertension in pregnancy is needed. The objective of this study is to assess the cost effectiveness of an innovative implementation strategy of the NVOG guidelines on hypertension including a computerised decision support system (BOS) compared to a common strategy of professional audit and feedback. A cluster randomised controlled trial with an economic evaluation alongside will be performed. Both pregnant women who develop severe hypertension or pre-eclampsia and professionals involved in the care for these women will participate. The main outcome measures are a combined rate of major maternal complications and process indicators extracted from the guidelines. A total of 472 patients will be included in both groups. For analysis, descriptive as well as regression techniques will be used. A cost effectiveness and cost utility analysis will be performed according to the intention-to-treat principle and from a societal perspective. Cost effectiveness ratios will be calculated using bootstrapping techniques.


Allergy in the Mediterranean area. III: Cross reactivity among Oleaceae pollens.
Pollens of closely related species often share common antigens and allergens although this is not always the case. Four species of Oleaceae pollens commonly found in the Mediterranean area were investigated for cross-reactivity: olive (Olea europaea), ash (Fraxinus exselsior), privet (Ligustrum vulgare) and Phillyrea angustifolia, a common bush. Twenty individual sera of patients allergic to Oleaceae were investigated for specific IgE antibodies against the four species of Oleaceae pollens. The results indicated a high degree of correlation between RAST titres of Oleaceae pollens but three gave a particular emphasis on one species only. Cross-reactivity among these four pollen species was sought by means of RAST inhibition, iso-electric focusing and tandem cross-immunoelectrophoresis. All tests revealed a high degree of cross reactivity although there is no total identity among these four pollen species.


Galectins as modulators of tumor progression in head and neck squamous cell carcinomas.
Head and neck squamous cell carcinomas (HNSCCs) remain a significant cause of morbidity worldwide. Biological therapies able to induce and/or upregulate antitumor immune responses could represent a complementary approach to conventional treatments for patients with HNSCC because, despite advances in surgery, radiotherapy, and chemotherapy, the overall survival rates for these patients have not changed over recent decades. Galectins are involved in the control of cell proliferation, cell death, and cell migration and in the modulation of various functions of the immune system. In this context, galectin-1 is known to protect HNSCCs from the immune system. The present review details the involvement of galectins in HNSCC biology and suggests a number of approaches to reduce the levels of expression of galectin-1 in HNSCCs, with the aim of improving the efficiency of HNSCC immunotherapy.


Phage-antibiotic combinations against multidrug-resistant Pseudomonas aeruginosa in in vitro static and dynamic biofilm models.
Biofilm-producing Pseudomonas aeruginosa infections pose a severe threat to public health and are responsible for high morbidity and mortality. Phage-antibiotic combinations (PACs) are a promising strategy for combatting multidrug-resistant (MDR), extensively drug-resistant (XDR), and difficult-to-treat P. aeruginosa infections. Ten MDR/XDR P. aeruginosa strains and five P. aeruginosa-specific phages were genetically characterized and evaluated based upon their antibiotic susceptibilities and phage sensitivities. Two selected strains, AR351 (XDR) and I0003-1 (MDR), were treated singly and in combination with either a broad-spectrum or narrow-spectrum phage, phage EM-T3762627-2_AH (EM), or 14207, respectively, and bactericidal antibiotics of five classes in biofilm time-kill analyses. Synergy and/or bactericidal activity was demonstrated with all PACs against one or both drug-resistant P. aeruginosa strains (average reduction: -Δ3.32 log10 CFU/cm2). Slightly improved ciprofloxacin susceptibility was observed in both strains after exposure to phages (EM and 14207) in combination with ciprofloxacin and colistin. Based on phage cocktail optimization with four phages (EM, 14207, E20050-C (EC), and 109), we identified several effective phage-antibiotic cocktails for further analysis in a 4-day pharmacokinetic/pharmacodynamic in vitro biofilm model. Three-phage cocktail, EM + EC + 109, in combination with ciprofloxacin demonstrated the greatest biofilm reduction against AR351 (-Δ4.70 log10 CFU/cm2 from baseline). Of remarkable interest, the addition of phage 109 prevented phage resistance development to EM and EC in the biofilm model. PACs can demonstrate synergy and offer enhanced eradication of biofilm against drug-resistant P. aeruginosa while preventing the emergence of resistance.


Efficacy and safety of daily use of tadalafil in treatment of patients with premature ejaculation: A randomised placebo-controlled clinical trial.
The study aimed to evaluate the efficacy and safety of the once-daily use of 5 mg tadalafil in the treatment of patients with premature ejaculation (PE). In a single-blind placebo-controlled clinical study, it was carried out on 100 patients with PE. All patients were randomised equally divided into two groups (50 patients each). Group 1 was given placebo in the form of oral multivitamin tablet once a day for 6 weeks. Group 2 was given 5 mg tadalafil once a day for 6 weeks. All patients were asked to complete Arabic Index of Premature Ejaculation (AIPE) before and after the treatment. This study showed that there were no statistically significant differences between patients in the placebo and tadalafil groups regarding the mean values of the seven questions and total scores of the AIPE questionnaire before treatment (p value >.05). The mean values of the seven questions and total scores of AIPE questionnaire in the tadalafil group were significantly higher than the placebo group after treatment (p value = .001). This study concluded that once-daily use of 5 mg tadalafil for 6 weeks was effective and well tolerated in the treatment of patients with PE.


Postmastectomy Radiation Therapy for Node-Negative Breast Cancer of 5 cm or Larger Tumors: A Multicenter Retrospective Analysis (KROG 20-03).
To evaluate the role of postmastectomy radiation therapy (PMRT) in patients with node-negative breast cancer of 5cm or larger tumors undergoing mastectomy. Medical records of 274 patients from 18 institutions treated with mastectomy between January 2000 and December 2016 were retrospectively reviewed. Among these, 202 patients underwent PMRT, while 72 did not. Two hundred and forty-one patients (88.0%) received systemic chemotherapy, and 172 (62.8%) received hormonal therapy. Patients receiving PMRT were younger, more likely to have progesterone receptor-positive tumors, and received adjuvant chemotherapy more frequently compared with those without PMRT (p <0.001, 0.018, and <0.001, respectively). Other characteristics were not significantly different between the two groups. With a median follow-up of 95 months (range, 1-249), there were 9 locoregional recurrences, and 20 distant metastases. The 8-year locoregional recurrence-free survival rates were 98.0% with PMRT and 91.3% without PMRT (p=0.133), and the 8-year disease-free survival (DFS) rates were 91.8% with PMRT and 73.9% without PMRT (p=0.008). On multivariate analysis incorporating age, histologic grade, lymphovascular invasion, hormonal therapy, chemotherapy, and PMRT, the absence of lymphovascular invasion and the receipt of PMRT were associated with improved DFS (p=0.025 and 0.009, respectively). Locoregional recurrence rate was very low in node-negative breast cancer of 5cm or larger tumors treated with mastectomy regardless of the receipt of PMRT. However, PMRT was significantly associated with improved DFS. Further investigation is needed to confirm these findings.


Docosahexaenoic acid inhibits the adhesion of flowing neutrophils to cytokine stimulated human umbilical vein endothelial cells.
The (n-3) PUFA, DHA, is widely thought to posses the ability to modulate the inflammatory response. However, its modes of interaction with inflammatory cells are poorly understood. In particular, there are limited data on the interactions of DHA with vascular endothelium, the cells that regulate the traffic of leukocytes from the blood into inflamed tissue. Using human umbilical vein endothelial cells (EC) cultured in a flow-based adhesion assay and activated with TNFα, we tested whether supplementing human umbilical vein EC with physiologically achievable concentrations of DHA would inhibit the recruitment of flowing neutrophils. DHA caused a dose-dependent reduction in neutrophil recruitment to the EC surface, although cells that became adherent were activated and could migrate across the human umbilical vein EC monolayer normally. Using EPA as an alternative supplement had no effect on the levels of neutrophil adhesion in this assay. Analysis of adhesion receptor expression by qPCR demonstrated that DHA did not alter the transcriptional activity of human umbilical vein EC. However, DHA did significantly reduce E-selectin expression at the human umbilical vein EC surface without altering the total cellular pool of this adhesion receptor. Thus, we have identified a novel mechanism by which DHA alters the trafficking of leukocytes during inflammation and demonstrate that this involves disruption of intracellular transport mechanisms used to present adhesion molecules on the surface of cytokine-stimulated EC.


The longitudinal course of cognitive impairment in schizophrenia: an examination of data from premorbid through posttreatment phases of illness.
Cognitive impairment is a core feature of schizophrenia that is present across the course of the illness. However, due to complexities of studying cognitive decline in patients prior to the onset of illness, the longitudinal course is not fully understood. The cognitive effects in patients with schizophrenia are robust, with a 1.5 to 2.5 standard deviation gap between patients and healthy controls on composite scores. People with schizophrenia manifest a prior history of cognitive impairment in the premorbid phases of the illness. Examination of school records suggests that children who will eventually develop schizophrenia begin school at a level of functioning that is a full grade behind their peers, with the gap increasing by the time they finish high school. Epidemiologic work suggests that there are both static cognitive impairments and developmental lags in these patients during childhood, well before the illness is fully manifest. Although there was initial promise of improved cognitive function with second-generation antipsychotic treatment, more recent studies have suggested no differences among antipsychotics, with the initial appearance of improvement very likely attributable to practice effects, inappropriate medication dosing, and poor study design. Two large, prominent studies evaluating first- and second-generation antipsychotics suggested that, although there was slight to modest improvement in cognitive function for all treatments, there were no differences among medications, regardless of the generation of the agents. In summary, patients who develop schizophrenia, on average, demonstrate cognitive impairment beginning as early as the first grade, with deterioration seen across school years. Further, these patients had substantial cognitive deficits after the initiation of psychosis. Finally, while antipsychotic treatment improves symptoms, antipsychotics have little impact on cognition, and there appear to be no differences in the degree of cognitive improvement between first- and second-generation agents.


Embedding Information into or onto Additively Manufactured Parts: A Review of QR Codes, Steganography and Watermarking Methods.
The paper gives a detailed review of the approaches adopted for embedding information into/onto additively manufactured parts. The primary purpose of this paper is to review all the techniques adopted for embedding information, highlight notable trends and improvements in these works, and provide design and manufacturing pipelines to realize most of these works. It classifies these approaches into four different categories and summarizes the works carried out in each field. It also compares all the results in textual and tabular forms and then gives a detailed conclusion of the best works in terms of application and effectiveness. The four categories discussed are 3D QR codes, 3D watermarking, steganography and nonclassified methods. Lastly, it discusses the future extensions and potential improvements in the field of embedding information, while exploring manufacturing technologies.


Cost-justification of a clinical pharmacist-managed anticoagulation clinic.
A cost-benefit evaluation of a clinical pharmacist-managed anticoagulation clinic (AC) was performed. Outpatient and hospital records were examined for 26 patients in the treatment group with an AC clinic and 26 patients in the control group. Therapeutic prothrombin times were maintained within the treatment group to a significantly greater extent than within the control group (p less than 0.001). The AC was successful in preventing hospitalizations resulting from hemorrhage or thromboembolism (p less than 0.01). The abnormal prothrombin times on admission in the control group correlated with hemorrhagic and thromboembolic admissions (p less than 0.05, p less than 0.005, respectively). Patients were hospitalized 3.22 days and .048 days per patient-treatment-year in the control and treatment groups, respectively. The net savings in reduced hospitalization costs per year in the treatment group was $211776. The benefit:cost ratio (B:C) was 6.55, suggesting the program is socially valuable. This clinical pharmacist-managed AC was effective in maintaining therapeutic prothrombin times, and reducing the incidence of hospitalization resulting from anticoagulation complications, and can be cost-justified based on a cost-benefit analysis.


Novel insecticidal toxins from nematode-symbiotic bacteria.
The current strategy of using transgenic crops expressing insecticidal protein toxins is placing increasing emphasis on the discovery of novel toxins, beyond those already derived from the bacterium Bacillus thuringiensis. Here we review the cloning of four insecticidal toxin complex (tc) encoding genes from a different bacterium Photorhabdus luminescens and of similar gene sequences from Xenorhabdus nematophilus. Both these bacteria occupy the gut of entomopathogenic nematodes and are released into the insect upon invasion by the nematode. In the insect the bacteria presumably secrete these insecticidal toxins, as well as a range of other antimicrobials, to establish the insect cadaver as a monocultural breeding ground for both bacteria and nematodes. In this review, the protein biochemistry and structure of the tc encoding loci are discussed in relation to their observed toxicity and histopathology. These toxins may prove useful as alternatives to those derived from B. thuringiensis for deployment in insect-resistant transgenic plants.


Development of in-vitro pulsatile flow generator for evaluating the performance of hemodialysis catheters.
Hemodialysis (HD) using an HD catheter is performed widely on renal failure patients. The catheter was evaluated using the recirculation ratio in pre-clinical status, which is a crucial index indicating its performance. However, pre-clinical in-vivo experiments have limitations: high cost, and ethical issues. Hence, computational and in-vitro methods have been developed as alternatives. However, computational methods require fluid dynamic knowledge, whereas in-vitro experiments are complicated and expensive. In this study, we developed a pulsatile flow generator to mimic blood flow achieving cost effectiveness and user convenience. The device used iterative learning control, achieving blood flow in the superior and inferior vena cava within a 3.3% error. Furthermore, the recirculation ratios were measured based on two insertion directions and two different external pipe materials to evaluate the catheter regarding patients' posture and blood vessel stiffness. The results provide a better understanding of cardiovascular device performance without complicated and costly pre-clinical tests.


A reappraisal of the definition and pathophysiology of the transient ischemic attack.
Recent data on the pathophysiology of brain ischemia obtained by neuroimaging methods and the new concept of transient ischemic attack (TIA) emergency have called for a redefinition of TIA. According to the new definition proposed by the TIA Working Group, TIA is a brief episode of neurological dysfunction caused by focal brain or retinal ischemia with clinical symptoms typically lasting less than one hour and without evidence of acute brain infarction. This new definition leads to a discussion on the duration of the neurological dysfunction and the availability of appropriate neuroimaging for all patients. It has been reported that the diffusion-weighted imaging abnormalities could be seen in TIA patients with durations of the neurological symptoms of less than 30 minutes, but they were not detected in 29% of patients with transient deficit lasting as much as 6 to 24 hours. Persisting perfusion abnormalities in TIA patients are also observed. Therefore, a cutoff period of any duration of TIA is inaccurate. From the pathophysiological viewpoint, TIA may be considered an ischemic penumbra of varied duration, which could proceed to cerebral infarction or reduce to benign oligemia. TIA, characterized as an ischemic penumbra, presents an ideal target for rapid reperfusion and neuroprotection. Follow-up perfusion imaging can guide and individualize its treatment.


Characterization of the processed form of a ubiquitous protein displaying a variable membrane organization in erythroid cells.
We recently identified a group of proteins that are present in all hematopoietic cells but are organized in the cell membrane of erythrocytes in a lineage-specific fashion (Blood 61:803, 1983). One of these polypeptides has a mol wt of approximately 37,000 (p37T) when translated in vitro from messenger RNA (mRNA) extracted from the erythroleukemic K562 cell line. The membrane-associated form of the p37 translation product has been analyzed in detail here. When detergent lysates prepared from biosynthetically labeled K562 cells were reacted with an antiserum containing anti-p37T antibodies, one of the proteins immunoprecipitated had a nominal mol wt of 36,000 to 37,000 (p37M). Several results suggest that this protein is homologous to the p37 translation product: (1) the protein, like the mRNA coding for the p37 translation product, was expressed in cell lines with diverse differentiated phenotypes; (2) the antigenic determinant(s) on p37M and p37T are oriented to the inner surface of the erythrocyte membrane while being oriented to the outer surface of erythroleukemic cells; and (3) one-dimensional peptide maps show homology between p37M and p37T. P37M does not appear to possess an N-terminal leader sequence that is proteolytically cleaved as the molecule is inserted into the membrane. In addition, p37M is not glycosylated.


Purified interleukin-1 (IL-1) from human monocytes stimulates acute-phase protein synthesis by rodent hepatocytes in vitro.
A universal component of inflammation is the increased synthesis of a series of plasma proteins (acute-phase proteins) by the liver. The postulated messenger of acute-phase protein induction is released by leucocytes at the site of inflammation and has been shown to co-purify with endogenous pyrogen or lymphocyte-activating factor. Interleukin-1, molecular weight 17,000, pI 6 X 8-7 X 2, was purified to homogeneity from adherent human blood monocytes by a combination of affinity chromatography, gel filtration and isoelectric focusing. We examined the direct effect of pure IL-1 on the induction of acute-phase protein synthesis in vitro using rat and mouse hepatocytes. IL-1 caused significant increased synthesis of alpha 1-acid glycoprotein and smaller increases in the synthesis of other acute-phase proteins, and significant decreased synthesis of albumin. The pattern of induction of acute-phase proteins differs from that seen with a separate 30,000 molecular weight hepatocyte-stimulating factor from human monocytes described previously. We conclude that human IL-1 is one of the mediators responsible for the acute-phase protein response of the liver in inflammation and can directly cause stimulation of specific gene expression in normal hepatocytes.


Principles and descriptions of design of skin flaps for use on the lower extremity.
Distinction must be made between tissue movability and tissue elasticity when considering tissue movement with the hope of predicting good results. Tissue movability is either inherent, as on the back of the thigh, or is created by undermining, back-cutting, or cutting out Bürow's triangles. The elasticity of skin is inherent everywhere and is difficult to accurately predict. Elastic tissue will gape when cut, whereas inelastic tissue will not. Finally, elastic tissue will rotate with little puckering but inelastic tissue will form large dog-ears when rotated. For these reasons, as soon as the decision has been made to use a flap it is important to "prethink" the dynamics of flap movement and decide which type of flap to use and what is to be accomplished. If consideration is given to which flap will best allow proper transfer of tissue and at the same time give the best cosmetic result, the task is usually simpler. By further considering the secondary movement of surrounding tissues (as well as the primary movement), the number of potential techniques and designs are greatly increased.


Responses of aquatic macrophyte cover and productivity to flooding variability on the Amazon floodplain.
Macrophyte net primary productivity (NPP) is a significant but understudied component of the carbon budget in large Amazonian floodplains. Annual NPP is determined by the interaction between stem elongation (vertical growth) and plant cover changes (horizontal expansion), each affected differently by flood duration and amplitude. Therefore, hydrological changes as predicted for the Amazon basin could result in significant changes in annual macrophyte NPP. This study investigates the responses of macrophyte horizontal expansion and vertical growth to flooding variability, and its possible effects on the contribution of macrophytes to the carbon budget of Amazonian floodplains. Monthly macrophyte cover was estimated using satellite imagery for the 2003-2004 and 2004-2005 hydrological years, and biomass was measured in situ between 2003 and 2004. Regression models between macrophyte variables and river-stage data were used to build a semiempirical model of macrophyte NPP as a function of water level. Historical river-stage records (1970-2011) were used to simulate variations in NPP, as a function of annual flooding. Vertical growth varied by a factor of ca. 2 over the simulated years, whereas minimum and maximum annual cover varied by ca. 3.5 and 1.5, respectively. Results suggest that these processes act in opposite directions to determine macrophyte NPP, with larger sensitivity to changes in vertical growth, and thus maximum flooding levels. Years with uncommonly large flooding amplitude resulted in the highest NPP values, as both horizontal expansion and vertical growth were enhanced under these conditions. Over the simulated period, annual NPP varied by ca. 1.5 (1.06-1.63 TgC yr(-1) ). A small increasing trend in flooding amplitude, and by extension NPP, was observed for the studied period. Variability in growth rates caused by local biotic and abiotic factors, and the lack of knowledge on macrophyte physiological responses to extreme hydrological conditions remain the major sources of uncertainty.


Coach-initiated motivational climate and cohesion in youth sport.
The general purpose of the present study was to examine the link between cohesion and motivational climate in youth sport. The first specific objective was to determine if relationships demonstrated in previous research with adult basketball and handball participants would be replicated in a younger sample and with a more heterogeneous set of sports. The second specific objective was to examine whether sources of athlete enjoyment moderate the relationships between motivational climate and cohesion. Athletes (N = 997; 532 girls and 465 boys; Mage = 15.26 +/- 1.20 years) completed measures pertaining to coach-initiated motivational climate, cohesion, and sources of enjoyment. Bivariate and canonical correlations revealed positive correlations between perceptions of a task-involving motivational climate and both task and social cohesion, while ego-involving motivational climate was negatively related. Cluster analyses suggested that individuals perceiving a low task-involving climate and high ego-involving climate perceived their teams as less cohesive. Finally, the degree to which participants derived enjoyment through other-referenced competency served as a moderator in the motivational climate-task cohesion relationship. Specifically, the relationship between task cohesion and motivational climate was more pronounced for those individuals who were less likely to derive enjoyment through other-referenced competency. Youth athletes' perceptions of coach-initiated motivational climate are related to cohesion. This relationship is, however, moderated by the degree to which athletes derive enjoyment through other-referenced competency. Motivational climate is an important variable to consider within team-building protocols intent on developing cohesion.


Evaluating the Accuracy of 2020 Census Block-Level Estimates in California.
In this study, we provide an assessment of data accuracy from the 2020 Census. We compare block-level population totals from a sample of 173 census blocks in California across three sources: (1) the 2020 Census, which has been infused with error to protect respondent confidentiality; (2) the California Neighborhoods Count, the first independent enumeration survey of census blocks; and (3) projections based on the 2010 Census and subsequent American Community Surveys. We find that, on average, total population counts provided by the U.S. Census Bureau at the block level for the 2020 Census are not biased in any consistent direction. However, subpopulation totals defined by age, race, and ethnicity are highly variable. Additionally, we find that inconsistencies across the three sources are amplified in large blocks defined in terms of land area or by total housing units, blocks in suburban areas, and blocks that lack broadband access.


Role of cholecystokinin and opioid peptides in control of food intake.
Of the many factors that influence food intake, there is strong evidence that opioid and CCK peptides, which stimulate feeding and elicit satiety, respectively, are important components that may act in concert to regulate energy balance. Cholecystokinin peptides have been isolated in both the brain and gastrointestinal tract, and changes in concentration in the brain and in plasma have been shown to vary with feeding. Peripherally injected CCK has been shown to elicit satiety in many species, including humans, an effect that may be mediated in the CNS via the vagus. In several species, most notably the sheep, direct injection into the CSF potently decreases food intake. Questions remaining regarding the role of CCK peptides in eliciting satiety include the sites and mechanisms of action. It is unknown whether CCK acts directly on receptors, indirectly on some other parameter, or as a neurotransmitter. Although opioid peptides have also been localized in portions of both the periphery and brain, a specific physiological role for their presence has not yet been determined. Opioid peptides from three families--endorphins, enkephalins, and dynorphins--have been shown to stimulate feeding in various species. They have been active at several opioid receptor types in the CNS, but there is limited evidence to suggest they affect food intake when administered peripherally. In contrast, peripheral injection of opiate antagonists has effectively decreased food intake, an observation that led to the original hypothesis that opioids were involved in the hunger component in the control of food intake and that excess concentrations might be involved in the development of obesity. An increasing body of evidence supports the concept that opioid and CCK peptides may interact to control food intake, but the evidence is more suggestive than conclusive.


The lack of analgesic use (oligoanalgesia) in small animal practice.
Oligoanalgesia is defined as failure to provide analgesia in patients with acute pain. Treatment of pain in emergencies, critical care and perioperatively may influence patient outcomes: the harmful practice of withholding analgesics occurs in teaching hospitals and private practices and results in severe physiological consequences. This article discusses the prevalence, primary causes, species and regional differences and ways to avoid oligoanalgesia in small animal practice. Oligoanalgesia may be addressed by improving education on pain management in the veterinary curriculum, providing continuing education to veterinarians and implementing pain scales.


Development and Validation of a Serum Metabolomic Signature for Endometrial Cancer Screening in Postmenopausal Women.
Endometrial carcinoma (EC) is the most commonly diagnosed gynecologic cancer. Its early detection is advisable because 20% of women have advanced disease at the time of diagnosis. To clinically validate a metabolomics-based classification algorithm as a screening test for EC. This diagnostic study enrolled 2 cohorts. A multicenter prospective cohort, with 50 cases (postmenopausal women with EC; International Federation of Gynecology and Obstetrics stage I-III and grade G1-G3) and 70 controls (no EC but matched on age, years from menopause, tobacco use, and comorbidities), was used to train multiple classification models. The accuracy of each trained model was then used as a statistical weight to produce an ensemble machine learning algorithm for testing, which was validated with a subsequent prospective cohort of 1430 postmenopausal women. The study was conducted at the San Giovanni di Dio e Ruggi d'Aragona University Hospital of Salerno (Italy) and Lega Italiana per la Lotta contro i Tumori clinic in Avellino (Italy). Data collection was conducted from January 2018 to February 2019, and analysis was conducted from January to March 2019. The presence or absence of EC based on evaluation of the blood metabolome. Metabolites were extracted from dried blood samples from all participants and analyzed by gas chromatography-mass spectrometry. A confusion matrix was used to summarize test results. Performance indices included sensitivity, specificity, positive and negative predictive values, positive and negative likelihood ratios, and accuracy. Confirmation or exclusion of EC in women with a positive test result was by means of hysteroscopy. Participants with negative results were followed up 1 year after enrollment to investigate the appearance of EC signs. The study population consisted of 1550 postmenopausal women. The mean (SD) age was 68.2 (11.7) years for participants with no EC in the training cohort, 69.4 (13.8) years for women with EC in the training cohort, and 59.7 (7.7) years for women in the validation cohort. Application of the ensemble machine learning to the validation cohort resulted in 16 true-positives, 2 false-positives, and 0 false-negatives, and it correctly classified more than 99% of samples. Disease prevalence was 1.12% (16 of 1430). In this study, dried blood metabolomic profile was used to assess the presence or absence of EC in postmenopausal women not receiving hormonal therapy with greater than 99% accuracy.


Neutrophil-lymphocyte ratio is prognostic in early stage resected small-cell lung cancer.
For selected early stage small cell lung cancer (SCLC), curative intent surgery is often performed. Previous studies, predominantly from East Asia, reported that high neutrophil to lymphocyte ratio (NLR), and platelet-lymphocyte ratio (PLR) correlate with poor prognosis in several types of tumors including SCLC. Our aim was to investigate the prognostic value of NLR and PLR in Caucasian patients with resected SCLC, as potential tool to select patients for multimodal treatment including surgery. Consecutive patients evaluated at three centers between 2000 and 2013 with histologically confirmed and surgically resected SCLC were retrospectively analyzed. NLR and PLR at diagnosis was used to categorize patients into "high" and "low" groups based on receiver operating curve analysis. Univariate and multivariate analyses were used to evaluate the impact of clinical and pathological characteristics on outcome. There were a total of 189 patients with a median age of 58 years, and the majority had stage I or II disease. We found a significant correlation between NLR and tumor stage (p = 0.007) and age (p = 0.038). Low NLR (LNLR) was associated with significantly longer overall survival, while PLR had no prognostic impact. There were significant associations between NLR and PLR but not with gender, vascular involvement, tumor necrosis, peritumoral inflammation, or tumor grade. Pre-operative LNLR may be a favorable prognostic factor in stage I-II SCLCs. PLR is not prognostic in this population. LNLR is easy to assess and can be integrated into routine clinical practice. Further prospective studies are needed to confirm these observations.


Decreased antibody response to influenza vaccination in kidney transplant recipients: a prospective cohort study.
Antibody response to the inactivated influenza vaccine is not well described in kidney transplant recipients administered newer, but commonly used, immunosuppression medications. We hypothesized that kidney transplant recipient participants administered tacrolimus-based regimens would have decreased antibody response compared with healthy controls. Prospective cohort study of 53 kidney transplant recipients and 106 healthy control participants during the 2006-2007 influenza season. All participants received standard inactivated influenza vaccine. Kidney transplant recipients administered tacrolimus-based regimens at a single academic medical center and healthy controls. Presence of kidney transplant. Proportion of participants achieving seroresponse (4-fold increase in antibody titer) and seroprotection (antibody titer > or = 1:32) 1 month after vaccination. Antibody titers before and 1 month after vaccination by means of hemagglutinin inhibition assays for influenza types A/H1N1, A/H3N2, and B. A smaller proportion of the transplantation group compared with the healthy control group developed the primary outcomes of seroresponse or seroprotection for all 3 influenza types at 1 month after vaccination. The response to influenza type A/H3N2 was statistically different; the transplantation group had 69% decreased odds of developing seroresponse (95% confidence interval, 0.16 to 0.62; P = 0.001) and 78% decreased odds of developing seroprotection (95% confidence interval, 0.09 to 0.53; P = 0.001) compared with healthy controls. When participants less than 6 months from the time of transplantation were considered, this group had a significantly decreased response to the vaccine compared with healthy controls. Decreased sample size, potential for confounders, outcome measure used is the standard but does not give information about vaccine efficacy. Kidney transplant recipients, especially within 6 months of transplantation, had diminished antibody response to the 2006-2007 inactivated influenza vaccine.


Enforcing a tobacco-free campus through an ambassador-based program: a phenomenology.
This phenomenology explored the experiences of those involved in a campus-based Ambassador program that enforced a tobacco-free policy at a public university in Montana. During the program's initial implementation (spring 2014), researchers observed and interviewed 21 Ambassadors and 19 policy violators. Researchers observed and recorded field notes of Ambassadors promoting the policy to the campus community and interacting with policy violators. Researchers invited Ambassadors and violators to be interviewed regarding their experiences with the program. Although Ambassadors enjoyed promoting the policy and sensed an increase in recognition by others, they also felt uncomfortable and had mixed feelings regarding their level of authority. Violators also felt uncomfortable, yet the majority described the experience with the program as positive. The findings from the study can be used to inform those considering the implementation of an Ambassador program on their campus.


Predominantly cystic central mucoepidermoid carcinoma developing from a previously diagnosed dentigerous cyst: case report and review of the literature.
Primary intraosseous mucoepidermoid carcinoma of the jawbones is an extremely rare malignant salivary gland tumour constituting 2-4.3% of all the reported mucoepidermoid carcinomas. We report a case of intraosseous mucoepidermoid carcinoma of the mandible in an 80-year old female patient developing from a previously diagnosed dentigerous cyst. An excisional biopsy was performed and the histopathological features confirmed low grade-cystic intraosseous mucoepidermoid carcinoma. The origin of central mucoepidermoid carcinoma could be suggested to be from the epithelial lining of previously diagnosed dentigerous cyst. Thus, emphasizing the need for careful examination of the entire excision specimen to rule out such neoplastic transformation of epithelial lining of odontogenic cyst and provide appropriate and effective treatment.


slan+ Monocytes Kill Cancer Cells Coated in Therapeutic Antibody by Trogoptosis.
Monocytes positive for 6-Sulfo LacNAc (slan) are a major subset of nonclassical CD14dimCD16+ monocytes in humans. We have shown that slan+ cells infiltrate lymphomas and elicit an antibody-dependent cellular cytotoxicity (ADCC) of neoplastic B cells mediated by the anti-CD20 therapeutic rituximab. Herein, by performing blocking experiments and flow cytometry analyses, as well as confocal microscopy and live-cell imaging assays, we extended the findings to other humanized antibodies and deciphered the underlying effector mechanism(s). Specifically, we show that, after coculture with target cells coated with anti-CD20 or anti-CD38, slan+ monocytes mediate trogocytosis, a cell-cell contact dependent, antibody-mediated process that triggers an active, mechanic disruption of target cell membranes. Trogocytosis by slan+ monocytes leads to a necrotic type of target cell death known as trogoptosis, which, once initiated, was partially sustained by endogenous TNFα. We also found that slan+ monocytes, unlike natural killer (NK) cells, mediate a direct ADCC with all types of anti-CD47 analyzed, and this was independent of their IgG isotype. The latter findings unveil a potentially relevant contribution by slan+ monocytes in mediating the therapeutic efficacy of anti-CD47 in clinical practice, which could be particularly important when NK cells are exhausted or deficient in number. Overall, our observations shed new light on the cytotoxic mechanisms exerted by slan+ monocytes in antibody-dependent tumor cell targeting and advance our knowledge on how to expand our therapeutic arsenal for cancer therapy.


Psychological well-being of Portuguese expatriates in Sub-Saharan Africa: a cross-sectional study.
Psychological health problems are highlighted among the most relevant disease patterns in expatriates. The purpose of this study was to determine the psychological well-being in Portuguese expatriates in Angola and Mozambique, considering the increasing expatriation wave. A cross-sectional self-administered web survey was conducted in a sample of 352 Portuguese civil expatriates in Angola and Mozambique. Clinically significant psychological distress was determined using General Health Questionnaire (GHQ)-12 and associated factors were studied using multiple logistic regression analysis. GHQ-12 items showed good internal consistency as reflected by the Cronbach's alpha. One-third of the screened expatriates corresponded to cases of clinically significant psychological distress. Age, country of birth, self-reported psychological symptoms and self-perception of general health in the previous 3 months were identified as independent variables associated with psychological distress. Increasing awareness and monitoring expatriates mental health should be in the health agenda, furthermore considering them a risk group in need of evidence-based mental health expatriate preparedness. The use of user-friendly validated tools, such as GHQ-12, allowing objective assessment and surveillance of these hard to reach populations should be reinforced.


Emergency Nursing Care of Patients With Novel Coronavirus Disease 2019.
Novel coronavirus disease 2019 is the disease caused by the novel coronavirus originally from Wuhan, China. Its pathophysiology is poorly understood, but it is known to be contagious and deadly. Multiple symptoms and complications from the disease have been described, with the most common complaints being respiratory. Nursing care of patients with novel coronavirus disease 2019 is largely supportive, but it should include a strong focus on mitigating the spread of infection to staff, other patients, and the community.


Recent advances in the ruthenium(ii)-catalyzed chelation-assisted C-H olefination of substituted aromatics, alkenes and heteroaromatics with alkenes via the deprotonation pathway.
The transition-metal-catalyzed chelation-assisted alkenylation at the inert C-H bond of aromatics with alkenes is one of the efficient methods to synthesize substituted vinylarenes in a highly regio- and stereoselective manner. Palladium, rhodium and ruthenium complexes are frequently used as catalysts for this type of transformation. The present review describes the recent advances in the ruthenium-catalyzed chelation-assisted alkenylation at the C-H bond of aromatics, alkenes and heteroaromatics with alkenes via the deprotonation pathway. Several directing groups including 2-pyridyl, carbonyl, amidine, amide, amine, imidate, sulphonic acid, triazole, cyano, oxazolidinone and hydontoin are widely used in the reaction. The scope, limitation and mechanistic investigation of the alkenylation reactions are discussed elaborately. This feature article includes all the reported ruthenium-catalyzed alkenylation reactions via the deprotonation pathway until the end of March 2017.


Poly(alkylcyanoacrylates) as biodegradable materials for biomedical applications.
This review considers the use of poly(alkylcyanoacrylates) (PACAs) as biomedical materials. We first present the different aspects of the polymerization of alkylcyanoacrylate monomers and briefly discuss their applications as skin adhesives, surgical glues and embolitic materials. An extensive review of the developments and applications of PACAs as nanoparticles for the delivery of drugs is then given. The methods of preparation of the nanoparticles are presented and considerations concerning the degradation, in vivo distribution, toxicity and cytotoxicity of the nanoparticles are discussed. The different therapeutic applications are presented according to the route of administration of the nanoparticles and include the most recent developments in the field.


Chemically modifying the mechanical properties of core-shell liquid metal nanoparticles.
Eutectic gallium-indium (EGaIn) is a room temperature liquid metal that can be readily fabricated into nanoparticles which naturally form a thin, passivating gallium oxide shell. These core-shell nanoparticles are of interest for a variety of stimuli-responsive applications, which often utilize physical deformation of the particles to release the molten, conductive payload from within the gallium oxide shell. In the present work, we introduce a variety of chemical strategies to produce EGaIn nanoparticles which exhibit a wide range of gallium oxide shell thicknesses. These chemically modified oxide thicknesses are then correlated to the core-shell liquid nanoparticles' mechanical properties by subjecting the particles to orthogonal characterization techniques; XPS for measurement of the gallium oxide shell thickness and nanoindentation for measurement of particle stiffness and elastic modulus. Additionally, nanoindentation is used to determine the onset of particle rupture and resultant conductivity. Ultimately, quantification of the relationships between chemical treatment and derivative mechanical properties in liquid metal nanoparticles will enable advanced applications of these colloids, such as in tailored self-healing and responsive electronic devices.


Altered-function p53 missense mutations identified in breast cancers can have subtle effects on transactivation.
Mutations of the sequence-specific master regulator p53 that alter transactivation function from promoter response elements (RE) could result in changes in the strength of gene activation or spectra of genes regulated. Such mutations in this tumor suppressor might lead to dramatic phenotypic changes and diversification of cell responses to stress. We have determined "functional fingerprints" of sporadic breast cancer-related p53 mutants, many of which are also associated with familial cancer proneness such as the Li-Fraumeni syndrome and germline BRCA1/2 mutant-associated cancers. The ability of p53, wild-type and mutants, to transactivate from 11 human target REs has been assessed at variable expression levels using a cellular, isogenomic yeast model system that allows for the rapid analysis of p53 function using a qualitative and a quantitative reporter. Among 50 missense mutants, 29 were classified as loss of function. The remaining 21 retained transactivation toward at least one RE. At high levels of galactose-induced p53 expression, 12 of 21 mutants that retain transactivation seemed similar to wild-type. When the level of galactose was reduced, transactivation defects could be revealed, suggesting that some breast cancer-related mutants can have subtle changes in transcription. These findings have been compared with clinical data from an ongoing neoadjuvant chemotherapy treatment trial for locally advanced breast tumors. The functional and nonfunctional missense mutations may distinguish tumors in terms of demographics, appearance, and relapse, implying that heterogeneity in the functionality of specific p53 mutations could affect clinical behavior and outcome.


Safety and immunogenicity of heterologous boosting with orally administered aerosolized bivalent adenovirus type-5 vectored COVID-19 vaccine and B.1.1.529 variant adenovirus type-5 vectored COVID-19 vaccine in adults 18 years and older: a randomized, double blinded, parallel controlled trial.
Vaccination strategies that can induce a broad spectrum immune response are important to enhance protection against SARS-CoV-2 variants. We conducted a randomized, double-blind and parallel controlled trial to evaluate the safety and immunogenicity of the bivalent (5×1010viral particles) and B.1.1.529 variant (5×1010viral particles) adenovirus type-5 (Ad5) vectored COVID-19 vaccines administrated via inhalation. A total of 451 eligible subjects aged 18 years and older who had been vaccinated with three doses of inactivated COVID-19 vaccines were randomly assigned to inhale one dose of either the B.1.1.529 variant Ad5 vectored COVID-19 vaccine (Ad5-nCoVO-IH group, N=150), bivalent Ad5 vectored COVID-19 vaccine (Ad5-nCoV/O-IH group, N=151), or Ad5 vectored COVID-19 vaccine (5×1010viral particles; Ad5-nCoV-IH group, N=150). Primary safety outcomes including adverse reactions reported by 37 (24.67%) participants in the Ad5-nCoVO-IH group, 28 (18.54%) in the Ad5-nCoV/O-IH group, and 26 (17.33%) in the Ad5-nCoV-IH group with mainly mild to moderate dry mouth, oropharyngeal pain, headache, myalgia, cough, fever and fatigue. No serious adverse events related to the vaccine were reported. All investigational vaccines were immunogenic, with the difference in the GMTs of neutralizing antibodies against Omicron BA.1 was statistically significant between Ad5-nCoV/O-IH (43.70) and Ad5-nCoV-IH (29.25) at 28 days after vaccination (P=0.0238). The seroconversion rates of neutralizing antibodies against BA.1 in Ad5-nCoVO-IH, Ad5-nCoV/O-IH, and Ad5-nCoV-IH groups were 56.00%, 59.60% and 48.67% with no significant difference among the groups. Overall, the investigational vaccines were demonstrated to be safe and well tolerated in adults, and was highly effective in inducing mucosal immunities in addition to humoral and cellular immune responses defending against SARS-CoV-2 variants.Trial registration: Chictr.org identifier: ChiCTR2200063996.


The Nature and Severity of Adverse Events in Select Outpatient Surgical Procedures in the Veterans Health Administration.
Research on adverse events (AEs) in outpatient surgery has been limited. As part of a Veterans Health Administration (VA) project on AE surveillance, we chart-reviewed selected outpatient surgical cases to characterize the nature and severity of AEs. We abstracted financial year 2012-2015 VA outpatient surgery cases selected with high (n = 1185) and low (n = 1072) likelihood of an AE based on postoperative health care utilization. The abstraction tool included established AE definitions and validated harm and severity scales. We found AEs in 608 high-likelihood (51%) and 126 low-likelihood outpatient surgical procedures (12%). Among 1010 unique AEs, the most common were wound issues (n = 261, 26%), urinary retention (23%), and urinary tract infections (12%). While 63% of all AEs involved minimal harm, 28% required hospitalization, and 9% were severely harmful including 8 AEs requiring intervention to sustain life and 2 deaths. Overall, 102 AEs (10%) required, at minimum, a repeat surgery to treat. Among VA outpatient surgical procedures selected based on likelihood of an AE, nearly 40% of identified events carried more than minimal patient harm, undermining the claim that outpatient surgery is relatively safe. Prevalent and preventable AEs such as wound dehiscence and urinary retention may be useful targets for quality improvement.


Identification and validation of stable reference genes for expression profiling of target genes in diverse ovine tissues.
Quantitative PCR (qPCR) is a widely-used technique for quantifying the expression of target genes across various tissues, as well as under different pathological and physiological conditions. One of the challenges associated with this method is the need to identify optimal reference genes (RGs) that maintain consistent expression levels under diverse experimental settings, thereby ensuring accurate biological interpretation. In this study, we conducted a thorough analysis of 18 candidate RGs (ACTB, BACH1, B2M, GAPDH, HMBS, HPRT1, PGK1, PPIA, PPIB, RPLP0, RPL19, RPS9, RPS15, RPS28, SDHA, TBP, UXT, and YWHAZ) across 10 ovine tissues (muscle, skin, kidney, liver, intestine, rumen, lung, testis, heart, and spleen) obtained from five individual sheep. We aimed to identify genes with stable expression across these tissues. A literature-based survey helped us shortlist candidate genes representing various functional classes from multiple livestock species. We employed four algorithms: geNorm, NormFinder, BestKeeper, and Delta Ct (ΔCt), to rank these genes based on their stability. A consistent trend in the rankings was observed across these different algorithms. RefFinder was then used for a comprehensive ranking, integrating the outputs from the various methods. ACTB, PPIB, BACH1, and B2M emerged as the most stable RGs, while RPS9, RPS15, and PGK1 displayed variable expression. We validated our findings through qPCR analysis of four target genes (ACTN2, CRYAB, DLK1, and TRIM54) in the skin samples from two different sheep breeds. Based on these results, we recommend ACTB, PPIB, BACH1, and B2M as reliable internal control genes for qPCR experiments involving diverse ovine tissues.


[Pathologichohistological skin changes in red-backed mice parasitized by larvae of the trombiculid mite, Neotromibcula pomeranzevi (Trombiculidae)].
During the feeding of N. pomeranzevi larvae on their natural hosgs, large-toothed redbacked voles (Clethronomys rufocanues Sandev.), in the skin of the latter are formed tubular structures or stylosomes characteristic of all trombiculids. During the formation of the stylosome the saliva of the mites and the interstitial fluid of the host are supposed to interact. The walls of the stylosome consist of two layers, are homogenous and do not include cellular elements. Around the stylosomes are formed the foci of necrosis and destruction of tissues. Behind the distal ends of the stylosomes are observed light interstitial cavities, which of then contain a great number of cellula relements of lymphoid and epithelioid nature. These cavities serve apparently as a reservoir of nutritive substratum for larvae. The feeding of trombiculids is accompanied by a reactive response of the connective tissue, the hyperemy of the superficial capillaries and the cellular infiltration of the affected area. The mass parasitism of larvae causes the ulceration of the host's skin and on its surface appear crusts from necrotic tissues.


Clinical features of mild cognitive impairment and dementia in a community: an update of the Osaki-Tajiri Project.
The borderline condition between normal aging and dementia is a major issue of concern for health policy planning because of an early intervention for possible prevention of dementia. Since 1988, the author has been involved in a community-based program on stroke, dementia, and bed-confinement prevention in Tajiri, northern Japan (the Osaki-Tajiri Project). As a part of the project, a cross-sectional study of aged patients with mild cognitive impairment in Tajiri was undertaken to investigate the clinical features of the condition, in addition to a longitudinal study to research its progression to dementia with possible risk factors. Impairment of the basic functions of attention and executive function was noted, as opposed to impairment in the cognitive domain itself. Magnetic resonance imaging (MRI) findings showed a pattern close to that of healthy persons in their 80s, rather than that of patients with cognitive deficit. The results of the longitudinal study showed more progression to dementia when the Clinical Dementia Rating (CDR) was 0.5 in domains other than memory. No effects of lifestyle, internal diseases or psychosocial intervention were confirmed. In progression to Alzheimer's disease, generally low cognitive function and general atrophy were involved, whereas frontal lobe function, atrophy of the frontal and temporal lobes, white matter changes and cerebral infarction were related to progression to vascular dementia. For health policy planning for dementia prevention, we consider that excessive dependence on primary prevention should be avoided; rather, secondary prevention, using the CDR, psychological testing and MRI are desirable.


Low levels of discriminant validity between self-report measures of self-esteem, shame, and self-criticism: Implications for the measurement of self-evaluation and creation of the Negative Self-Evaluation Scale.
This article examined the discriminant and convergent validity of commonly used self-report measures of self-criticism, self-esteem, and shame. A confirmatory factor analysis (CFA) using multiple self-report measures of each construct showed low levels of discriminant validity between self-reported self-esteem, shame, and self-criticism and instead demonstrated correspondingly high levels of shared variance. However, bifactor analyses on the items across each measure suggested that self-report measures of self-esteem, shame, and self-criticism may contain distinct characteristics that are underrepresented in current measures of each construct. Based on the factor loadings in item-level bifactor analyses, a new measure, the Negative Self-Evaluation Scale (NSES), was constructed to improve the assessment of the unique characteristics of shame, self-esteem, and self-criticism. Implications for current and future practices concerning the measurement of each construct are discussed. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Exon 3 splicing and mutagenesis identify residues influencing cell surface density of heterologously expressed silkworm (Bombyx mori) glutamate-gated chloride channels.
Glutamate-gated chloride channels (GluCls) mediate fast inhibitory neurotransmission in invertebrate nervous systems. Insect GluCls show alternative splicing, and, to determine its impact on channel function and pharmacology, we isolated GluCl cDNAs from larvae of the silkworm (Bombyx mori). We show that six B. mori glutamate-gated chloride channel variants are generated by splicing in exons 3 and 9 and that exons 3b and 3c are common in the brain and third thoracic ganglion. When expressed in Xenopus laevis oocytes, the three functional exon 3 variants (3a, b, c) all had similar EC50 values for l-glutamate and ivermectin (IVM); however, Imax (the maximum l-glutamate- and IVM-induced response of the channels at saturating concentrations) differed strikingly between variants, with the 3c variant showing the largest l-glutamate- and IVM-induced responses. By contrast, a partial deletion detected in exon 9 had a much smaller impact on l-glutamate and IVM actions. Binding assays using [(3)H]IVM indicate that diversity in IVM responses among the GluCl variants is mainly due to the impact on channel assembly, altering receptor cell surface numbers. GluCl variants expressed in HEK293 cells show that structural differences influenced Bmax but not Kd values of [(3)H]IVM. Domain swapping and site-directed mutagenesis identified four amino acids in exon 3c as hot spots determining the highest amplitude of the l-glutamate and IVM responses. Modeling the GluCl 3a and 3c variants suggested that three of the four amino acids contribute to intersubunit contacts, whereas the other interacts with the TM2-TM3 linker, influencing the receptor response.


Auxin Exposure Disrupts Feeding Behavior and Fatty Acid Metabolism in Adult Drosophila.
The ease of genetic manipulation in Drosophila melanogaster using the Gal4/UAS system has been beneficial in addressing key biological questions. Current modifications of this methodology to temporally induce transgene expression require temperature changes or exposure to exogenous compounds, both of which have been shown to have detrimental effects on physiological processes. The recently described auxin-inducible gene expression system (AGES) utilizes the plant hormone auxin to induce transgene expression and is proposed to be the least toxic compound for genetic manipulation, with no obvious effects on Drosophila development and survival in one wild-type strain. Here we show that auxin delays larval development in another widely-used fly strain, and that short- and long-term auxin exposure in adult Drosophila induces observable changes in physiology and feeding behavior. We further reveal a dosage response to adult survival upon auxin exposure, and that the recommended auxin concentration for AGES alters feeding activity. Furthermore, auxin fed male and female flies exhibit a significant decrease in triglyceride levels and display altered transcription of fatty acid metabolism genes. Although fatty acid metabolism is disrupted, auxin does not significantly impact adult female fecundity or progeny survival, suggesting AGES may be an ideal methodology for studying limited biological processes. These results emphasize that experiments using temporal binary systems must be carefully designed and controlled to avoid confounding effects and misinterpretation of results.


Nonrecurrent laryngeal nerves and their clinical significance.
The anomalous position of a nonrecurrent laryngeal nerve predisposes the nerve to injury during thyroidectomy and to compression by a thyroid mass. We present three cases in which a seemingly benign thyroid mass traumatized a nonrecurrent laryngeal nerve resulting in either vocal cord paralysis or a vague pressure sensation over the larynx. Some of these patients feel as if they need to clear a foreign body and present with a chronic cough. Normally the nerve is protected from thyroid masses as it passes through the tracheoesophageal groove. In all three patients, surgical excision of the thyroid mass and release of the nerve resulted in recovery of the nerve and resolution of the symptoms. We have found that small, benign, or otherwise asymptomatic lesions of the thyroid gland have a greater tendency to cause vocal cord paralysis in patients with nonrecurrent laryngeal nerves. The surgeon must always be aware of the possibility of the presence of a nonrecurrent laryngeal nerve.


Computational investigation of dust mite allergens in a realistic human nasal cavity.
Aim: Inhaled allergens from house dust mite (HDM) are a major source of allergic disease such as allergic rhinitis and asthma. It has been a challenge to properly evaluate health risks caused by HDM related allergens including mite bodies, eggs and fecal pellets. This paper presents a numerical study on particle deposition of dust mite allergens in a human nasal cavity. Materials and methods: A realistic nasal cavity model was reconstructed from CT scans and a Computational Fluid Dynamics analysis of steady airflow was simulated. The discrete phase model was used to trace particle trajectories of three dust mite related particles. Results: The flow and particle model were validated by comparing with nasal resistance measurement and previous literature respectively. Aerodynamic characteristics and deposition of dust mite allergens in the nasal cavity were analyzed under different breathing conditions including rest and exercising conditions. Conclusions: The numerical results revealed the roles of different nasal cavity regions in filtering various types of dust mite allergens with consideration of breathing conditions.


NLRX1 negatively modulates type I IFN to facilitate KSHV reactivation from latency.
Kaposi's sarcoma-associated herpesvirus (KSHV) is a herpesvirus that is linked to Kaposi's sarcoma (KS), primary effusion lymphoma (PEL) and multicentric Castleman's disease (MCD). KSHV establishes persistent latent infection in the human host. KSHV undergoes periods of spontaneous reactivation where it can enter the lytic replication phase of its lifecycle. During KSHV reactivation, host innate immune responses are activated to restrict viral replication. Here, we report that NLRX1, a negative regulator of the type I interferon response, is important for optimal KSHV reactivation from latency. Depletion of NLRX1 in either iSLK.219 or BCBL-1 cells significantly suppressed global viral transcription levels compared to the control group. Concomitantly, fewer viral particles were present in either cells or supernatant from NLRX1 depleted cells. Further analysis revealed that upon NLRX1 depletion, higher IFNβ transcription levels were observed, which was also associated with a transcriptional upregulation of JAK/STAT pathway related genes in both cell lines. To investigate whether IFNβ contributes to NLRX1's role in KSHV reactivation, we treated control and NLRX1 depleted cells with a TBK1 inhibitor (BX795) or TBK1 siRNA to block IFNβ production. Upon BX795 or TBK1 siRNA treatment, NLRX1 depletion exhibited less inhibitory effects on reactivation and infectious virion production, suggesting that NLRX1 facilitates KSHV lytic replication by negatively regulating IFNβ responses. Our data suggests that NLRX1 plays a positive role in KSHV lytic replication by suppressing the IFNβ response during the process of KSHV reactivation, which might serve as a potential target for restricting KSHV replication and transmission.


Acquired factor IX inhibitor in a patient with adenocarcinoma of the colon.
The association of an acquired inhibitor to factor IX and adenocarcinoma of the sigmoid colon is reported here for the first time. Unlike the lupus anticoagulant, the inhibitor identified did not exhibit marked prolongation of the clotting time using a highly diluted thromboplastin nor was the kinetic behavior of the inhibitor-factor IX interaction time-dependent. Administration of prednisone was associated with the return of the activated partial thromboplastin time to normal values, at which time the tumor was excised and corticosteroids discontinued with no evidence of subsequent factor IX inhibitor activity.


Understanding the complex relationship between multidimensional poverty and depressive symptoms among young South Africans: A cross-sectional study.
We use the Global Multidimensional Poverty Index (MPI) to explore how different dimensions of poverty more directly linked to young people are associated with depressive symptoms among South African youth. Data came from the 2017 wave of the nationally-representative National Income Dynamics Study (NIDS) in South Africa. We focused on a sample of 15-24-year-olds whose depressive symptoms were assessed using an adapted version of the 10-item Centre for Epidemiological Studies Depression Scale. We examine how individual dimensions and indicators of the MPI relate to depression, in comparison to more conventional measures, including household income, subjective social standing, overcrowding and personal assets. Cross-sectional analyses were adjusted for clustering to account for sampling design. The MPI index was not associated with probable depression (OR = 1.02, 95 % CI 0.81-1.29). Only lack of access to the labour market emerged as a key individual dimension associated with probable depression (OR = 5.29, 95 % CI 1.70-16.47), a relationship driven by an increased odds for those not in employment, education or training. Lack of household assets, living in an informal dwelling and lower perceived social standing were also associated with increased odds for depression. No gender differences were noted. The study is cross-sectional and not suitable to examine the causal nature of the association between multidimensional poverty and depression. Poverty dimensions that measure youth's access to employment or training have a strong association with depression. Further research is needed to assess whether improved access to employment or training contributes to improving mental health among young South Africans.


Etiological heterogeneity and clinical characteristics of metopic synostosis: Evidence from a tertiary craniofacial unit.
Metopic synostosis (MS) accounts for approximately 10-15% of all craniosynostosis and is etiologically heterogeneous. This study aimed to examine the causes of MS, as observed in a tertiary craniofacial unit. We reviewed the case notes of 110 children with a diagnosis of MS, attending the craniofacial unit in Oxford between 1991 and 2008. Our results showed 38 children (38/110 or 34.6%) who had at least one additional structural abnormality or had a known syndromic diagnosis were classed as having syndromic MS. Chromosomal abnormalities were noted in 8/38 (21.4%) children: mosaic marker chromosome 2, 9p deletion (2/8), 11q deletion, 12pter deletion and duplication of 15q25 with other additional chromosomal abnormalities (3/8). Other syndromic diagnoses included Silver-Russell syndrome and Greig cephalopolysyndactyly. Prenatal exposure to sodium valproate (VPA) was noted in 8/110 children (7.8%), with the dose of the VPA being >or=1,000 mg/day in all cases. Other prenatal exposures reported in this study were: maternal diabetes (6/110), enoxaparin for hypercoagulable state (1/110), and thyroxine (1/110). The majority of patients (72/110 or 65.4%) had nonsyndromic MS. Speech delay was present in 11 children with nonsyndromic MS (11/72 or 15.3%) and 10 children with syndromic MS (10/38 or 26.3%). We conclude that approximately two-thirds of all MS is nonsyndromic. Prenatal exposure to VPA is a common cause of MS. Maternal diabetes, not previously linked to MS, was noted in 5.5% of cases. Chromosomal abnormalities account for about 6% of MS. An increased risk of speech delay is seen with both the syndromic and nonsyndromic forms.


Suppressing activity of tributyrin on hepatocarcinogenesis is associated with inhibiting the p53-CRM1 interaction and changing the cellular compartmentalization of p53 protein.
Hepatocellular carcinoma (HCC), an aggressive and the fastest growing life-threatening cancer worldwide, is often diagnosed at intermediate or advanced stages of the disease, which substantially limits therapeutic approaches for its successful treatment. This indicates that the prevention of hepatocarcinogenesis is probably the most promising approach to reduce both the HCC incidence and cancer-related mortality. In previous studies, we demonstrated a potent chemopreventive effect of tributyrin, a butyric acid prodrug, on experimental hepatocarcinogenesis. The cancer-inhibitory effect of tributyrin was linked to the suppression of sustained cell proliferation and induction of apoptotic cell death driven by an activation of the p53 apoptotic signaling pathway. The goal of the present study was to investigate the underlying molecular mechanisms linked to tributyrin-mediated p53 activation. Using in vivo and in vitro models of liver cancer, we demonstrate that an increase in the level of p53 protein in nuclei, a decrease in the level of cytoplasmic p53, and, consequently, an increase in the ratio of nuclear/cytoplasmic p53 in rat preneoplastic livers and in rat and human HCC cell lines caused by tributyrin or sodium butyrate treatments was associated with a marked increase in the level of nuclear chromosome region maintenance 1 (CRM1) protein. Mechanistically, the increase in the level of nuclear p53 protein was associated with a substantially reduced binding interaction between CRM1 and p53. The results demonstrate that the cancer-inhibitory activity of sodium butyrate and its derivatives on liver carcinogenesis may be attributed to retention of p53 and CRM1 proteins in the nucleus, an event that may trigger activation of p53-mediated apoptotic cell death in neoplastic cells.


Clinical Efficacy of Azithromycin as an Adjunctive Therapy to Non-Surgical Periodontal Treatment of Periodontitis: A Systematic Review and Meta-Analysis.
Over the years, numerous trials have sufficiently documented benefits of adjuvant administration of antibiotics in combination to Scaling and Root Planing (SRP) in treatment of periodontitis. Nevertheless there are justified doubts in respect to antibiotics administration with regard to increasing development of resistances. The aim of this review study and meta-analysis was to verify a possible benefit of Azithromycin (AZM) as an alternative adjuvant antibiotic in combination with SRP. Electronic databases (Pubmed) were searched from the earliest point of record until May 2015. A hand search was also conducted. The variables considered in this respect were Probing Depth (PD), Clinical Attachment Level (CAL), Bleeding on Probing (BOP) and SRP. A meta-analysis containing trials comparing SRP as a sole therapy with SRP and administration of AZM was produced once all of the data obtained had been reviewed. Six randomised clinical trials were accepted into the review and the meta-analysis following the process of trial selection. Meta-analysis revealed larger overall effects by the systemic administration of AZM compared with SRP on their own in terms of PD, CAL, BOP, Plaque-Index and total bacterial load reduction after three and six months. The data used in this study revealed that the systemic administration of AZM is beneficial compared with SRP on its own for the treatment of chronic periodontitis.


Comparison of vessel contrast measured with a scanning-beam digital x-ray system and an image intensifier/television system.
Vessel contrast was measured in the fluoroscopic images produced by a scanning-beam digital x-ray (SBDX) system and an image intensifier/television (II/TV) based system. The SBDX system electronically scans a series of pencil x-ray beams across the patient, each of which is directed at a distant small-area detector array. The reduction in detected scatter achieved with this geometry was expected to provide an increase in image contrast. Vessel contrast was evaluated from images of a phantom containing iodinated tubes. The vessels were inserted into an acrylic stack to provide a patient-mimicking scattering medium. Vessel diameter ranged from 0.3 to 3.1 mm. Images were acquired at 100 kVp with the SBDX and II/TV systems and averaged to reduce x-ray noise. The II/TV system was operated in the 6-in. image intensifier mode with an anti-scatter grid. The increase in contrast in the SBDX images, expressed as a ratio of the measured SBDX and II/TV contrasts, ranged from 1.63 to 1.79 for individual vessels. This agreed well with a prediction of the contrast improvement ratio for this experiment, based on measurements of the scatter fraction, object-plane line spread functions, and consideration of the source spectrum and detector absorption properties. The predicted contrast improvement ratio for SBDX relative to II/TV images was 1.62 to 1.77.


[Morphofunctional evaluation of the results of reconstructive operations of gastroenteroanastomosis in peptic ulcer].
The mucosa of the gastric stump, abducting and adducting loops of small intestine in patients with peptic ulcers of gastrointestinal anastomosis has been examined with scanning, transmission electron and light microscopy, morphometry, and the study of secretion at various periods after reconstructive surgery combined with vagotomy. An increase of the number of chief and parietal cells in the gastric stump fundal glands, increase of the thickness of the mucosa due to the deepening of the crypts of the adducting and abducting loops of the small intestine, and alteration of the mucosal microrelief of the gastric stump and small intestine are noted in peptic ulcers of gastrointestinal anastomosis especially near the site of anastomosis. This is accompanied by an increase of mucosal secretory activity. Reconstructive operations combined with vagotomy cause a reduction in chief and parietal cells number, smoothing of the mucosal microrelief and decrease of gastric secretory activity.


Change of diagnosis of post-traumatic stress disorder related to compensation-seeking.
To explore the change in the diagnosis of posttraumatic stress disorder (PTSD) related to the implementation of the new national regulation on compensation-seeking by war veterans in Croatia. The study included 225 compensation-seeking war veterans who were psychiatrically assessed and diagnosed on three different occasions. The first diagnosis was made by a local psychiatrist when a veteran sought psychiatric help for the first time. The second psychiatric diagnosis was established during the veteran's psychiatric treatment, and the third one was made by an independent expert team in charge of the psychiatric assessment for compensation purposes. The expert examination included structured diagnostic procedure and analysis of military service data. The diagnoses established on three different points were compared. There were significant differences between the diagnoses of mental disorders made at three different occasions in compensation-seeking war veterans. Six different diagnostic categories of mental disorders were confirmed. The diagnosis changed in 134 (59.5%) out of 225 veterans, mainly in the categories of PTSD and personality changes due to catastrophic experience, during their psychiatric treatment in the 2000-2002 period, when the new regulation for compensation was implemented. PTSD diagnosis remained unchanged at all three psychiatric assessments in only 7.5% of the veterans, whereas the diagnosis of a mental disorder remained unchanged in 37 (16.4%) veterans. Experts' confirmation of PTSD or PTSD comorbid with other mental disorders positively correlated with the number of hospitalizations. Inconsistencies in the diagnosis of PTSD could be related to the different diagnostic criteria and the course of illness. Psychiatric examination for compensation purposes should be independent and integrate all relevant data for making a complete assessment. Compensation-seeking policy, represented by new regulations, could be a source of bias in diagnostic outcome.


Decline and Pronounced Regional Disparities in Medical Cocaine Usage in the United States.
Background: Cocaine is a stimulant and Schedule II drug used as a local anesthetic and vasoconstrictor. Objective: This descriptive study characterized medical cocaine use in the United States. Methods: Retail drug distribution data from 2002 to 2017 were extracted for each state from the Drug Enforcement Administration, which reports on medical, research, and analytical chemistry use. The percentage of buyers (pharmacies, hospitals, and providers) was obtained. Use per state, corrected for population, was determined. Available cross-sectional data on cocaine use as reported by the Medicare and Medicaid programs for 2013-2017 and electronic medical records were examined. Results: Medical cocaine use decreased by -62.5% from 2002 to 2017. Hospitals accounted for 84.9% and practitioners for 9.9% of cocaine distribution in 2017. The number of pharmacies carrying cocaine dropped by -69.4%. The percentages of hospitals, practitioners, and pharmacies that carried cocaine in 2017 were 38.4%, 2.3%, and 0.3%, respectively. There was a 7-fold difference in 2002 (South Dakota, 76.1 mg/100 persons; Delaware, 10.1 mg/100 persons). Relative to the average state in 2017, those reporting the highest values (Montana, 20.1; North Dakota, 24.1 mg/100 persons) were significantly elevated. Cocaine use within the Medicare and Medicaid programs was negligible. Cocaine use within the Geisinger system was rare from 2002 to 2007 (<4 orders/100 000 patients per year) but increased to 48.7 in 2018. Conclusion and Relevance: If these pharmacoepidemiological patterns continue, licit cocaine may soon become a historical relic. The pharmacology and pharmacotherapeutics education of health care providers may need to be adjusted accordingly.


Merkel cell carcinoma of unknown primary site.
Merkel cell carcinoma is an uncommon and aggressive primary neuroendocrine skin malignancy which mostly affects the extremities and the head and neck region of elderly patients. Merkel cell carcinoma occurs with increased frequency in sun-exposed areas, in individuals exposed to arsenic and in immunosuppressed patients. Many patients with Merkel cell carcinoma present with other malignancies, mainly skin cancers. Characteristic features are frequent recurrences and regional and distant metastases. Mortality rates range from 20 to 65 per cent. The mainstay of treatment is surgery, with wide local excision, and adjuvant radiotherapy is usually administered. Merkel cell carcinoma of unknown primary site is rare, and the majority of the few cases described have not been from head and neck areas. We present a case of Merkel cell carcinoma of unknown primary site, with upper neck and distant metastases.


Carbon dioxide laser polishing of fused silica surfaces for increased laser-damage resistance at 1064 nm.
Mechanically polished fused silica surfaces were heated with continuous-wave CO(2) laser radiation. Laser-damage thresholds of the surfaces were measured with 1064-nm 9-nsec pulses focused to small spots and with large-spot, 1064-nm, 1-nsec irradiation. A sharp transition from laser-damage-prone to highly laser-damage-resistant took place over a small range in CO(2) laser power. The transition to high damage resistance occurred at a silica surface temperature where material softening began to take place as evidenced by the onset of residual strain in the CO(2) laser-processed part. The small-spot damage measurements show that some CO(2) laser-treated surfaces have a local damage threshold as high as the bulk damage threshold of SiO(2). On some CO(2) laser-treated surfaces, large-spot damage thresholds were increased by a factor of 3-4 over thresholds of the original mechanically polished surface. These treated parts show no obvious change in surface appearance as seen in bright-field, Nomarski, or total internal reflection microscopy. They also show little change in transmissive figure. Further, antireflection films deposited on CO(2) laser-treated surfaces have thresholds greater than the thresholds of antireflection films on mechanically polished surfaces.


Effects of GH replacement therapy on thyroid volume and nodule development in GH deficient adults: a retrospective cohort study.
Despite the well-known effects of GH/IGF1 signaling on the thyroid, few data are available on the risk of developing nodular goiter in hypopituitary subjects during GH replacement therapy (GHRT). We aimed to define the effects of GH therapy on thyroid volume (TV) and nodular growth. The records of 96 subjects (47 males and 49 females, median age 48 years) with GH deficit (GHD) were investigated. Seventy also had central hypothyroidism (CH). At the time of our retrospective evaluation, median treatment duration was 5 years. Pre-treatment TV was smaller in GHD patients than in healthy subjects (P=0.030). During GH treatment, TV significantly increased (P=0.016 for the entire group and P=0.014 in euthyroid GHD patients). Before starting GH therapy, 17 patients harbored thyroid nodules. During GH therapy, nodule size increased slightly in seven patients, and new thyroid nodules occurred in nine patients. Among the 79 patients without pre-existing thyroid nodules, 17 developed one or more nodules. There was no difference in the prevalence of CH in GHD patients with or without thyroid nodules (P=0.915; P=0.841, when patients with pre-therapy nodular goiter were excluded), the main predictor for nodule development being serum IGF1 (P=0.038). GHRT is associated with TV's increase in GHD patients. Thyroid nodules developed in 27% of patients, mainly in relation to pre-therapy IGF1 levels, independently of normal or impaired TSH stimulation.


Orexins acting at native OX(1) receptor in colon cancer and neuroblastoma cells or at recombinant OX(1) receptor suppress cell growth by inducing apoptosis.
Screening of 26 gut peptides for their ability to inhibit growth of human colon cancer HT29-D4 cells grown in 10% fetal calf serum identified orexin-A and orexin-B as anti-growth factors. Upon addition of either orexin (1 microM), suppression of cell growth was total after 24 h and >70% after 48 or 72 h, with an EC(50) of 5 nm peptide. Orexins did not alter proliferation but promoted apoptosis as demonstrated by morphological changes in cell shape, DNA fragmentation, chromatin condensation, cytochrome c release into cytosol, and activation of caspase-3 and caspase-7. The serpentine G protein-coupled orexin receptor OX(1)R but not OX(2)R was expressed in HT29-D4 cells and mediated orexin-induced Ca(2+) transients in HT29-D4 cells. The expression of OX(1)R and the pro-apoptotic effects of orexins were also indicated in other colon cancer cell lines including Caco-2, SW480, and LoVo but, most interestingly, not in normal colonic epithelial cells. The role of OX(1)R in mediating apoptosis was further demonstrated by transfecting Chinese hamster ovary cells with OX(1)R cDNA, which conferred the ability of orexins to promote apoptosis. A neuroblastoma cell line SK-N-MC, which expresses OX(1)R, also underwent growth suppression and apoptosis upon treatment with orexins. Promotion of apoptosis appears to be an intrinsic property of OX(1)R regardless of the cell type where it is expressed. In conclusion, orexins, acting at native or recombinant OX(1)R, are pro-apoptotic peptides. These findings add a new dimension to the biological activities of these neuropeptides, which may have important implications in health and disease, in particular colon cancer.


Toxic injury to isolated hepatocytes is not dependent on extracellular calcium.
Freshly isolated hepatocytes from phenobarbital-treated rats were incubated in the presence or absence of extracellular calcium with three differently acting liver cell toxins, namely carbon tetrachloride, bromobenzene, and ethylmethanesulfonate. In the absence of extracellular calcium these three compounds were far more toxic to the cells than in its presence. This result is inconsistent with the hypothesis that an influx of extracellular calcium is required as the final step in toxic liver cell injury.