The development of prosocial behaviour in children and adolescents: a twin study.
Childhood psychopathology is associated with both high and low levels of prosocial behaviour. It has been proposed that the development of prosocial behaviour shows emerging and consolidating individual differences as children grow older. The influences on these individual differences have not previously been examined in children and adolescents using multiple raters in a genetically informative design. Twin data from 682 families based on parent and teacher reports were used to examine the genetic and environmental influences on prosocial behaviour in 5-16-year-olds. Effects of sex, age and rater were examined. There were no significant differences in the magnitude of genetic and environmental influence on male and female prosocial behaviour. Declining common environment and increasing genetic influences were seen with age. This emerged as a trend in parent data and reached statistical significance in teacher data. When parent and teacher data were examined together in a rater bias model significant bias acting on the parent ratings emerged, in keeping with previous discrepancies between parental and observational measures. There was overlap in the phenotype rated by parents and teachers, with a highly heritable common underlying phenotype. The influences on the distribution of prosocial behaviour in children and adolescents show declining shared environmental and increasing genetic influences with age. Parental assessments of prosocial behaviour show significantly higher scores than teacher reports and whilst there is overlap in the phenotype rated by parents and teachers, parents show significant bias in their ratings.


Mass, energy and material balances of SRF production process. Part 2: SRF produced from construction and demolition waste.
In this work, the fraction of construction and demolition waste (C&D waste) complicated and economically not feasible to sort out for recycling purposes is used to produce solid recovered fuel (SRF) through mechanical treatment (MT). The paper presents the mass, energy and material balances of this SRF production process. All the process streams (input and output) produced in MT waste sorting plant to produce SRF from C&D waste are sampled and treated according to CEN standard methods for SRF. Proximate and ultimate analysis of these streams is performed and their composition is determined. Based on this analysis and composition of process streams their mass, energy and material balances are established for SRF production process. By mass balance means the overall mass flow of input waste material stream in the various output streams and material balances mean the mass flow of components of input waste material stream (such as paper and cardboard, wood, plastic (soft), plastic (hard), textile and rubber) in the various output streams of SRF production process. The results from mass balance of SRF production process showed that of the total input C&D waste material to MT waste sorting plant, 44% was recovered in the form of SRF, 5% as ferrous metal, 1% as non-ferrous metal, and 28% was sorted out as fine fraction, 18% as reject material and 4% as heavy fraction. The energy balance of this SRF production process showed that of the total input energy content of C&D waste material to MT waste sorting plant, 74% was recovered in the form of SRF, 16% belonged to the reject material and rest 10% belonged to the streams of fine fraction and heavy fraction. From the material balances of this process, mass fractions of plastic (soft), paper and cardboard, wood and plastic (hard) recovered in the SRF stream were 84%, 82%, 72% and 68% respectively of their input masses to MT plant. A high mass fraction of plastic (PVC) and rubber material was found in the reject material stream. Streams of heavy fraction and fine fraction mainly contained non-combustible material (such as stone/rock, sand particles and gypsum material).


Role of modern radiotherapy in treating skin cancer.
Radiotherapy is an important modality in the treatment of non-melanoma skin cancers. While the majority of patients will be adequately treated without the need for radiotherapy, there are factors that may favour a recommendation for radiotherapy. The established roles of modern radiotherapy include the definitive, adjuvant and palliative settings. It is important that clinicians treating skin cancers have an understanding and awareness of these roles. The aim of this article is to present an overview of radiotherapy in the current Australian setting.


Evidence of an increased pathogenic footprint in the lingual microbiome of untreated HIV infected patients.
Opportunistic oral infections can be found in over 80% of HIV + patients, often causing debilitating lesions that also contribute to deterioration in nutritional health. Although appreciation for the role that the microbiota is likely to play in the initiation and/or enhancement of oral infections has grown considerably in recent years, little is known about the impact of HIV infection on host-microbe interactions within the oral cavity. In the current study, we characterize modulations in the bacterial composition of the lingual microbiome in patients with treated and untreated HIV infection. Bacterial species profiles were elucidated by microarray assay and compared between untreated HIV infected patients, HIV infected patients receiving antiretroviral therapy, and healthy HIV negative controls. The relationship between clinical parameters (viral burden and CD4+ T cell depletion) and the loss or gain of bacterial species was evaluated in each HIV patient group. In untreated HIV infection, elevated viremia was associated with significantly higher proportions of potentially pathogenic Veillonella, Prevotella, Megasphaera, and Campylobacter species in the lingual microbiome than observed in healthy controls. The upsurge in the prevalence of potential pathogens was juxtaposed by diminished representation of commensal Streptococcus and Veillonella species. Colonization of Neisseria flavescens was lower in the lingual microbiome of HIV infected patients receiving antiretroviral therapy than in uninfected controls. Our findings provide novel insights into the potential impact of HIV infection and antiretroviral therapy on the community structure of the oral microbiome, and implicate potential mechanisms that may increase the capacity of non-commensal species to gain a stronger foothold.


Interference with human immunodeficiency virus (HIV) replication by CD8+ T cells in peripheral blood leukocytes of asymptomatic HIV carriers in vitro.
A long asymptomatic period is one of the characteristics of human immunodeficiency virus (HIV) infection, despite its fatal consequences. Antiviral defense in HIV-infected individuals controls viral replication during this period. In the present study, we demonstrate that peripheral blood leukocytes (PBL) of asymptomatic HIV-1 carriers, following exogenous HIV-1 infection in vitro, do not support viral replication. These cells do not produce detectable amounts of reverse transcriptase or accumulate unintegrated proviral DNA. This is a striking contrast to the behavior of HIV-1-infected PBL of seronegative individuals, which produce large amounts of RT and unintegrated DNA. Such resistance to HIV-1 replication is not seen in PBL of patients with advanced disease. Since the binding of HIV-1 to CD4 molecule is not impaired in PBL of asymptomatic carriers, the interference with HIV replication must occur after the stage of virus binding. PBL lose their resistance when CD8+ lymphocytes are removed. In addition, these PBL are not resistant to an exogenous infection with HIV-2. These observations suggest that certain populations of CD8+ lymphocytes of asymptomatic HIV-1 carriers operate on the target cells in PBL to block viral replication in an HIV-1-specific manner. Such CD8+ lymphocyte-mediated interference with HIV replication could play an important role in the maintenance of the period of disease latency.


Osthole/borneol thermosensitive gel via intranasal administration enhances intracerebral bioavailability to improve cognitive impairment in APP/PS1 transgenic mice.
Alzheimer's disease (AD) poses a significant threat to the global elderly population. Traditional Chinese medicine (TCM) has been widely utilized in the treatment of AD. Osthole, a bioactive ingredient classified as an "emperor" in many TCM formulas, has been demonstrated to effectively alleviate AD symptoms. However, its low bioavailability in the brain has limited its clinical application. This study aimed to increase the intracerebral bioavailability of osthole by using borneol as a "courier," based on the classical "Emperor-Minister-Assistant-Courier" model, and to investigate the enhanced pharmacological performance of osthole on AD. Results indicated that a suitable in situ thermosensitive gel matrix for intranasal administration mixed with osthole and borneol consists of P407 at 20%, P188 at 7%, and PEG300 at 6%. The concentration of osthole in the cerebrospinal fluid increased almost tenfold after intranasal administration of osthole/borneol compared to oral administration. Mechanisms showed that borneol as a "courier" opened up intercellular space and loosened the tight junctions of the nasal mucosa by suppressing ZO-1 and occludin expression, thereby expediting the nose-to-brain route and guiding osthole as "emperor" to its target in the brain. Osthole assisted by borneol demonstrated significantly improved efficiency in suppressing cleaved caspase-3 expression, increasing the Bcl-2/Bax ratio, improving T-SOD and catalase expression, reducing malondialdehyde levels, inhibiting neuron apoptosis, and decreasing Aβ levels by inhibiting BACE1 expression to alleviate cognitive impairment in APP/PS1 mice compared to osthole alone. Overall, our study demonstrated that the intracerebral bioavailability of osthole profoundly improved with intranasal administration of osthole/borneol and provided a wider application of TCM for AD treatment with higher intracerebral bioavailability.


Quantitative proteomic analysis using a MALDI quadrupole time-of-flight mass spectrometer.
We describe an approach to the quantitative analysis of complex protein mixtures using a MALDI quadrupole time-of-flight (MALDI QqTOF) mass spectrometer and isotope coded affinity tag reagents (Gygi, S. P.; et al. Nat. Biotechnol. 1999, 17, 994-9.). Proteins in mixtures are first labeled on cysteinyl residues using an isotope coded affinity tag reagent, the proteins are enzymatically digested, and the labeled peptides are purified using a multidimensional separation procedure, with the last step being the elution of the labeled peptides from a microcapillary reversed-phase liquid chromatography column directly onto a MALDI sample target. After addition of matrix, the sample spots are analyzed using a MALDI QqTOF mass spectrometer, by first obtaining a mass spectrum of the peptides in each sample spot in order to quantify the ratio of abundance of pairs of isotopically tagged peptides, followed by tandem mass spectrometric analysis to ascertain the sequence of selected peptides for protein identification. The effectiveness of this approach is demonstrated in the quantification and identification of peptides from a control mixture of proteins of known relative concentrations and also in the comparative analysis of protein expression in Saccharomyces cerevisiae grown on two different carbon sources.


Unveiling the high reactivity of cyclohexynes in [3 + 2] cycloaddition reactions through the molecular electron density theory.
[3 + 2] cycloaddition (32CA) reactions of cyclohexyne, a cyclic strained acetylene, with methyl azide and methoxycarbonyl diazomethane have been studied within the Molecular Electron Density Theory (MEDT) at the MPWB1K/6-311G(d) computational level. These 32CA reactions, which take place through a one-step mechanism involving highly asynchronous transition state structures, proceed with relatively low activation enthalpies of 6.0 and 4.3 kcal mol-1, respectively, both reactions being strongly exothermic. The reactions are initiated by the creation of a pseudoradical center at one of the two acetylenic carbons of cyclohexyne with a very low energy cost, 1.0 kcal mol-1, which promotes the easy formation of the first C-N(C) single bond in the adjacent acetylenic carbon. This scenario is completely different from that of the 32CA reaction involving non-strained but-2-yne; thus, strain in cyclohexyne triggers a high reactivity as a consequence of its unusual electronic structure at the ground state. Finally, the experimental regioselectivity of the 32CA reactions involving 3-alkoxy-cyclohexyne derivatives is correctly explained within MEDT.


Therapeutic ultrasound-enhanced immune checkpoint inhibitor therapy.
Immune checkpoint inhibitors (ICIs) are designed to reinvigorate antitumor immune responses by interrupting inhibitory signaling pathways and promoting the immune-mediated elimination of malignant cells. Although ICI therapy has transformed the landscape of cancer treatment, only a subset of patients achieve a complete response. Focused ultrasound (FUS) is a noninvasive, nonionizing, deep penetrating focal therapy that has great potential to improve the efficacy of ICIs in solid tumors. Five FUS modalities have been incorporated with ICIs to explore their antitumor effects in preclinical studies, namely, high-intensity focused ultrasound (HIFU) thermal ablation, HIFU hyperthermia, HIFU mechanical ablation, ultrasound-targeted microbubble destruction (UTMD), and sonodynamic therapy (SDT). The enhancement of the antitumor immune responses by these FUS modalities demonstrates the great promise of FUS as a transformative cancer treatment modality to improve ICI therapy. Here, this review summarizes these emerging applications of FUS modalities in combination with ICIs. It discusses each FUS modality, the experimental protocol for each combination strategy, the induced immune effects, and therapeutic outcomes.


High-level expression and characterization of a novel cutinase from Malbranchea cinnamomea suitable for butyl butyrate production.
Butyl butyrate has been considered as a promising fuel source because it is a kind of natural ester which can be converted from renewable and sustainable lignocellulosic biomass. Compared with the conventional chemical methods for butyl butyrate production, the enzymatic approach has been demonstrated to be more attractive, mainly owing to the mild reaction conditions, high specificity, low energy consumption, and environmental friendliness. Cutinases play an important role in the butyl butyrate production process. However, the production level of cutinases is still relatively low. Thus, to identify novel cutinases suitable for butyl butyrate synthesis and enhance their yields is of great value in biofuel industry. A novel cutinase gene (McCut) was cloned from a thermophilic fungus Malbranchea cinnamomea and expressed in Pichia pastoris. The highest cutinase activity of 12, 536 U/mL was achieved in 5-L fermentor, which is by far the highest production for a cutinase. McCut was optimally active at pH 8.0 and 45 °C. It exhibited excellent stability within the pH range of 3.0-10.5 and up to 75 °C. The cutinase displayed broad substrate specificity with the highest activity towards p-nitrophenyl butyrate and tributyrin. It was capable of hydrolyzing cutin, polycaprolactone, and poly(butylene succinate). Moreover, McCut efficiently synthesized butyl butyrate with a maximum esterification efficiency of 96.9% at 4 h. The overall structure of McCut was resolved as a typical α/β-hydrolase fold. The structural differences between McCut and Aspergillus oryzae cutinase in groove and loop provide valuable information for redesign of McCut. These excellent features make it useful in biosynthesis and biodegradation fields. A novel cutinase from M. cinnamomea was identified and characterized for the first time. High-level expression by P. pastoris is by far the highest for a cutinase. The enzyme exhibited excellent stability and high esterification efficiency for butyl butyrate production, which may make it a good candidate in biofuel and chemical industries.


Design, statistical analysis and reporting standards for test accuracy studies for infectious diseases in animals: Progress, challenges and recommendations.
The quality of diagnostic accuracy studies (DAS) for infectious diseases of animals has improved over the last 20 years because of international educational efforts, use of design and reporting standards to guide researchers and test developers, and acceptance of the use of latent class models to account for imperfect reference tests. In this review, we focus on measurement of diagnostic sensitivity and specificity as a measure of clinical validity, describe the leadership role of the World Organisation of Animal Health (OIE) in setting standards for test validation in the context of fitness-for-purpose, and describe how design and reporting quality have facilitated the increased use of systematic reviews and meta-analysis of DAS. Ongoing challenges for design, conduct, analysis and reporting of DAS are identified; and we make recommendations for improvements in these areas for OIE-listed and non-listed infectious diseases.


Concordance of a decision algorithm and multidisciplinary team meetings for patients with liver cancer-a study protocol for a randomized controlled trial.
Multidisciplinary team meetings (MDMs), also known as tumor conferences, are a cornerstone of cancer treatments. However, barriers such as incomplete patient information or logistical challenges can postpone tumor board decisions and delay patient treatment, potentially affecting clinical outcomes. Therapeutic Assistance and Decision algorithms for hepatobiliary tumor Boards (ADBoard) aims to reduce this delay by providing automated data extraction and high-quality, evidence-based treatment recommendations. With the help of natural language processing, relevant patient information will be automatically extracted from electronic medical records and used to complete a classic tumor conference protocol. A machine learning model is trained on retrospective MDM data and clinical guidelines to recommend treatment options for patients in our inclusion criteria. Study participants will be randomized to either MDM with ADBoard (Arm A: MDM-AB) or conventional MDM (Arm B: MDM-C). The concordance of recommendations of both groups will be compared using interrater reliability. We hypothesize that the therapy recommendations of ADBoard would be in high agreement with those of the MDM-C, with a Cohen's kappa value of ≥ 0.75. Furthermore, our secondary hypotheses state that the completeness of patient information presented in MDM is higher when using ADBoard than without, and the explainability of tumor board protocols in MDM-AB is higher compared to MDM-C as measured by the System Causability Scale. The implementation of ADBoard aims to improve the quality and completeness of the data required for MDM decision-making and to propose therapeutic recommendations that consider current medical evidence and guidelines in a transparent and reproducible manner. The project was approved by the Ethics Committee of the Charité - Universitätsmedizin Berlin. The study was registered on ClinicalTrials.gov (trial identifying number: NCT05681949; https://clinicaltrials.gov/study/NCT05681949 ) on 12 January 2023.


Palladium(ii)-catalyzed γ-selective hydroarylation of alkenyl carbonyl compounds with arylboronic acids.
A catalytic γ-selective syn-hydroarylation of alkenyl carbonyl compounds using arylboronic acids has been developed using a substrate directivity approach with a palladium(ii) catalyst. This method tolerates a wide range of functionalized (hetero)arylboronic acids and a variety of substitution patterns on the alkene. Preliminary mechanistic studies suggest that transmetalation is rate-limiting.


Deregulation of collagen phagocytosis in aging human fibroblasts: effects of integrin expression and cell cycle.
Intracellular degradation of collagen by phagocytosis in fibroblasts is essential for physiological remodeling of the extracellular matrix in a wide variety of connective tissues but imbalances between degradation and synthesis can lead to loss of tissue collagen. As aging is associated with loss of dermal and periodontal collagen and with increased lysomomal enzyme content in fibroblasts, we examined the regulation of collagen phagocytosis by integrin expression and the cell cycle in an in vitro fibroblast aging model. Two different fibroblast lines (CL1; CL2) at the fourth subculture were passaged up to replicative senescence to model aging processes in vitro. Cells were incubated with collagen-coated or BSA-coated green fluorescent beads for 3 h to assess alpha 2 beta 1-integrin-mediated or nonspecific phagocytosis, respectively. Single-cell suspensions were stained with DAPI and sulforhodamine 101 to separate cycling G1 and noncycling G0 cells. Staining for alpha 2-integrin, bead internalization, and bivariate analyses of DNA/protein content were measured by three-color flow cytometry. Serum deprivation was used to induce increases in the proportion of G0 cells. For G1 cells, the proportion of collagen phagocytic cells was > 50% for all passages and collagen beads were internalized > 5-fold more frequently than BSA beads. In contrast, G0 cells with diploid DNA content but low protein content exhibited greatly reduced phagocytic capacity (< 10% of cells internalized collagen or BSA beads), the number of beads per cells was 4-fold less, and alpha 2 integrin expression was very low compared to G1 cells. The proportion of collagen phagocytic cells and the proportion of alpha 2-integrin-positive cells increased with transit through the cell cycle. At higher passage numbers mean cell volume and cytoplasmic granularity were reduced approximately 30% but at replicative senescence cells with large surface area and subdiploid DNA predominated. The proportion of collagen and BSA phagocytic G1 cells increased 1.5- and 5-fold, respectively, and the number of beads per cell increased < 3-fold. However, surface alpha 2-integrin staining remained unchanged. These data indicate that the collagen and nonspecific internalization pathways were greatly unregulated, independent of cell cycle phase, and that cellular aging in vitro strongly influences the specificity and rate of phagocytic processes in fibroblasts. We suggest that age-related loss of collagen in connective tissues undergoing turnover may be a manifestation of a deregulated increase of collagen phagocytosis in which the net loss of degraded collagen exceeds new synthesis.


Initial Investigation of the Psychometric Properties of the Limited Prosocial Emotions Questionnaire (LPEQ).
Callous-unemotional traits, which include lack of remorse or guilt, callousness/lack of empathy, unconcern about performance, and shallow/deficient affect, were included as a specifier of conduct disorder in the current (fifth) edition of the Diagnostic and Statistical Manual of Mental Disorders under the rubric Limited Prosocial Emotions (LPEs). The purpose of this study was to examine the psychometric properties of a new rating scale, the Limited Prosocial Emotions Questionnaire (LPEQ). Caregivers (n = 1,050) of children (Mage = 8.42, SD = 2.31) completed the LPEQ and other measures. Results provide support for a single factor model of the LPEQ, with measurement invariance supported across child and informant sex. Both the reliability and validity of the LPEQ as a measure of LPEs were also well supported. Children identified with LPE had significantly greater average impairment and need for treatment relative to children without LPE. Children with conduct problems (i.e., conduct disorder or oppositional defiant disorder), as well those without conduct problems, had significantly more impairment if they were identified as having LPE. Our findings fit with the mounting evidence of the clinical utility of assessing LPEs in children. Future research should look to replicate our findings in clinical samples of youth.


Simultaneous detection of IFN-gamma and IL-4 mRNAs using RT-PCR and time-resolved fluorometry.
Time-resolved fluorometry was applied in the detection of RT-PCR amplified mRNAs for the Th1 and Th2 cell-derived cytokines interferon gamma (IFN-gamma) and interleukin (IL-)4, respectively. RNA stimulated cells was reverse transcribed and the cDNAs for the cytokine mRNAs and the constantly expressed beta-actin (beta-ACT) mRNA were simultaneously amplified in one multiplex PCR reaction. The PCR conditions were optimized to minimize mutual inhibition of individual amplifications. One of the PCR primers in each primer pair was biotinylated, and the PCR products were captured onto streptavidin-coated microtitre plates. The three PCR products were detected with three different lanthanide labelled target-specific probes in solution hybridization. IFN-gamma, IL-4 and beta-ACT were detected with europium (Eu), terbium (Tb) and samarium (Sm) labelled probes, respectively, using time-resolved fluorometry. Small cell numbers used in microtitre plate cultures were sufficient to detect cytokine messages after mitogen stimulation. This sequence-based method provides a sensitive, specific, fast and nonisotopic alternative to conventional blotting and hybridisation with radioactive probes. In addition, the multiplex fluorogenic dye detection facilitates relative quantification of target mRNAs.


Mechanisms of human smooth muscle cell proliferation and transplant vasculopathy induced by HLA class I antibodies: in vitro and in vivo studies.
Vascular smooth muscle cells (SMC) play an important role in the pathophysiology of transplant vasculopathy (TV), a major cause of late death in patients receiving an organ transplant. In this review we describe the proliferative effect in vitro and in vivo of HLA class I antibodies on human SMC. We have developed an experimental model using segments of human mesenteric arteries transplanted in the position of the infrarenal aorta in immunodeficient mice (SCID/beige). Weekly injections of transplanted mice with a monoclonal antibody towards HLA class I provoked typical lesions of TV after 6 weeks in the human graft while transplanted mice receiving an irrelevant antibody did not develop any significant lesion. In vitro, the anti-HLA antibodies were mitogenic to SMC and we showed that they activate a stress-induced signaling pathway implicating matrix metalloproteinases (MMP) and neutral sphingomyelinase 2 (nSMase-2). The proliferative effect of anti-HLA antibodies could be blocked by pharmacological inhibitors or by siRNA. Administration of pharmacological inhibitors diminished the development of TV in grafted mice injected with anti-HLA antibodies demonstrating an important role of the MMP/nSMase-2 pathway in antibody-induced TV. This observation opens new perspectives for the management of TV in clinical settings.


Incidence and clinical characteristics of periocular infantile hemangiomas.
To report the incidence, demographics, and clinical findings among a population-based cohort of children with periocular infantile hemangiomas. The medical records of all patients (<19 years of age) diagnosed as having periocular infantile hemangiomas while residing in Olmsted County, Minnesota, from January 1, 1965, through December 31, 2004, were retrospectively reviewed. Forty-three children were diagnosed as having periocular infantile hemangiomas during the 40-year period, yielding an incidence of 5.4 per 100,000 individuals younger than 19 years (95% CI, 3.8-7.1) or a birth prevalence of 1 in 1586 live births. Thirty children (70%) were female (P<.001). There was a history of maternal infertility in approximately 1 in 5 children and premature birth in 1 in 8 children. Twenty-six children (61%) had other abnormalities, including secondary hemangiomas in 9 (21%). Forty-one patients (95%) had unilateral disease, and 37 hemangiomas (86%) were located on the upper eyelid. In this population-based study, periocular infantile hemangiomas occurred in 1 in 1586 live births and were most prevalent on the unilateral upper eyelid of white female patients. Prevalent associations included maternal infertility and premature birth. Other abnormalities, including secondary hemangiomas in 1 in 5 children, were common in this cohort.


Antibiotic combinations for the empiric treatment of the febrile neutropenic patient.
Empiric antimicrobial therapy in the granulocytopenic patient has traditionally consisted of a combination of an aminoglycoside plus a cephalosporin or aminoglycoside plus an antipseudomonal penicillin. These combinations have been proven reasonably effective and only moderately toxic. In some centers trimethoprim-sulfamethoxazole combined with an aminoglycoside or antipseudomonal penicillin has been used, again with good efficacy and minimal toxicity. The newer extended spectrum penicillins and cephalosporins has made possible two-drug combinations with aminoglycosides that provide double coverage of nearly all of the common bacterial pathogens while concurrently providing synergy against most of these same organisms. Further, it is now possible to have a double beta-lactam combination of a penicillin and cephalosporin which has the advantages of broad spectrum by both antimicrobials and, at times, synergistic activity but without the ototoxicity and nephrotoxicity of aminoglycosides. The double beta-lactam combination would be especially useful in the neutropenic patient with established renal insufficiency or a history of previous courses of aminoglycoside therapy. Care should be taken in choosing such a regimen, however, because some combinations demonstrate antagonism. There is increasing data to indicate that patients with profound (less than 100 microliters), persistent (greater than 14 days) granulocytopenia and concurrent gram-negative bacteremia respond best if treated with two active, bactericidal antibiotics, preferably synergistic in action. The potential for synergistic activity of the drugs selected should be, therefore, carefully considered. The duration of therapy should be adjusted according to the severity of infection, the response of the infection to antimicrobial therapy, and rate of recovery of the patient's granulocyte count.


[Anatomical reconstruction of posterolateral complex in treatment of multi-ligament injury of knees].
To evaluate the short-term effectiveness after static anatomical reconstruction of posterolateral complex (PLC) in the treatment of traumatic multi-ligament injury of the knee. Between June 2007 and July 2011, 23 cases of multi-ligament injury of the knee were treated. There were 15 males and 8 females with an average age of 41 years (range, 19-56 years). The injury was caused by traffic accident in 9 cases, sprain in 7 cases, bruise in 3 cases, and falling from height in 4 cases. The time between injury and operation was 13-78 days (mean, 32 days). The results of posterior drawer test and Lachman test were positive, and all cases complicated by varus and external rotation instability. The Lysholm score of the knee was 43.4 +/- 5.7. According to International Knee Documentation Committee (IKDC) scoring, all were rated as grade D. According to Fanelli typing, all were classified as type C. The X-ray films showed that load-induced posterior motion of the knee was (13.3 +/- 4.2) mm; the lateral joint space was (15.1 +/- 2.4) mm. Anterior cruciate ligament/posterior cruciate ligament and PLC were reconstructed simultaneously with auto-semitendinosus, gracilis tendon, and allogeneic tendon. All incisions healed by first intention, and no complication occurred. All patients were followed up 12-56 months (mean, 28 months). At last follow-up, the results of posterior drawer test and Lachman test were negative; 3 cases had varus instability, and 2 cases had external rotation instability. The Lysholm score of the knee was 85.6 +/- 16.7, showing significant difference when compared with preoperative score (t=-11.469, P=0.000). According to IKDC scoring, 7 cases were rated as grade A, 12 as grade B, and 4 as grade C; significant difference was found when compared with preoperative value (Z=4.285, P=0.000). The load-induced posterior motion of the knee was (5.1 +/- 4.4) mm, the lateral joint space was (3.2 +/- 2.8) mm, showing significant differences when compared with preoperative ones (P < 0.05). In the treatment of traumatic multi-ligament injury of the knee, the anatomical reconstruction of the PLC using auto-semitendinosus, gracilis tendon, or allogeneic tendon can obtain good short-term effectiveness.


Effectiveness of Time-Restricted Eating with Caloric Restriction vs. Caloric Restriction for Weight Loss and Health: Meta-Analysis.
Time-restricted eating (TRE) is an increasingly popular dietary strategy for weight loss. Recent studies suggest that combining TRE with caloric restriction (CR) may have more favorable effects on both physical and biochemical aspects when compared with CR alone. Therefore, we performed a meta-analysis to compare the effects of TRE with CR vs. CR alone on anthropometric and biochemical measures in overweight or obese adults. We reviewed articles from PubMed, Web of science, EMBASE, and the Cochrane Library published before 25 May 2023. The meta-analysis incorporated data from seven randomized controlled trials of nine interventions, with a total of 231 participants in the TRE with CR group and 227 participants in the CR-only group. Data were analyzed using RewMan version 5.4.1. All results in our meta-analysis were described as mean difference (MD) with 95% confidence interval (Cl). Results showed that TRE with CR compared to CR alone resulted in significantly greater reductions in body weight (MD: -2.11 kg, 95% CI: -2.68 kg to -1.54 kg, p = < 0.00001, I2 = 42%), body fat mass (MD: -0.75 kg, 95% CI: -1.35 kg to -0.16 kg, p = 0.01; I2 = 0%), and waist circumference (MD: -1.27 cm, 95% CI: -2.36 cm to -0.19 cm, p = 0.02, I2 = 0%), while no additional impact of TRE in combination with CR in comparison to CR on serum biochemical parameters were found. Our results suggest that the improvement in biochemical parameters are mainly caused by CR, while improvements in anthropometric parameters are further enhanced by TRE.


A reverse taxonomic approach to assess macrofaunal distribution patterns in abyssal Pacific polymetallic nodule fields.
Heightened interest in the exploitation of deep seafloor minerals is raising questions on the consequences for the resident fauna. Assessing species ranges and determination of processes underlying current species distributions are prerequisites to conservation planning and predicting faunal responses to changing environmental conditions. The abyssal central Pacific nodule belt, located between the Clarion and Clipperton Fracture Zones (CCZ), is an area prospected for mining of polymetallic nodules. We examined variations in genetic diversity and broad-scale connectivity of isopods and polychaetes across the CCZ. Faunal assemblages were studied from two mining claims (the eastern German and French license areas) located 1300 km apart and influenced by different productivity regimes. Using a reverse taxonomy approach based on DNA barcoding, we tested to what extent distance and large-scale changes in environmental parameters lead to differentiation in two macrofaunal taxa exhibiting different functions and life-history patterns. A fragment of the mitochondrial gene Cytochrome Oxidase Subunit 1 (COI) was analyzed. At a 97% threshold the molecular operational taxonomic units (MOTUs) corresponded well to morphological species. Molecular analyses indicated high local and regional diversity mostly because of large numbers of singletons in the samples. Consequently, variation in composition of genotypic clusters between sites was exceedingly large partly due to paucity of deep-sea sampling and faunal patchiness. A higher proportion of wide-ranging species in polychaetes was contrasted with mostly restricted distributions in isopods. Remarkably, several cryptic lineages appeared to be sympatric and occurred in taxa with putatively good dispersal abilities, whereas some brooding lineages revealed broad distributions across the CCZ. Geographic distance could explain variation in faunal connectivity between regions and sites to some extent, while assumed dispersal capabilities were not as important.


Disruption of MAPK1 expression in the ERK signalling pathway and the RUNX1‑RUNX1T1 fusion gene attenuate the differentiation and proliferation and induces the growth arrest in t(8;21) leukaemia cells.
The t(8;21) translocation is one of the most frequent chromosome abnormalities associated with acute myeloid leukaemia (AML). This abberation deregulates numerous molecular pathways including the ERK signalling pathway among others. Therefore, the aim of the present study was to investigate the gene expression patterns following siRNA‑mediated suppression of RUNX1‑RUNX1T1 and MAPK1 in Kasumi‑1 and SKNO‑1 cells and to determine the differentially expressed genes in enriched biological pathways. BeadChip microarray and gene ontology analysis revealed that RUNX1‑RUNX1T1 and MAPK1 suppression reduced the proliferation rate of the t(8;21) cells with deregulated expression of several classical positive regulator genes that are otherwise known to enhance cell proliferation. RUNX1‑RUNX1T1 suppression exerted an anti‑apoptotic effect through the overexpression of BCL2, BIRC3 and CFLAR genes, while MAPK1 suppression induced apopotosis in t(8;21) cells by the apoptotic mitochondrial changes stimulated by the activity of upregulated TP53 and TNFSF10, and downregulated JUN gene. RUNX1‑RUNX1T1 suppression supported myeloid differentiation by the differential expression of CEBPA, CEBPE, ID2, JMJD6, IKZF1, CBFB, KIT and CDK6, while MAPK1 depletion inhibited the differentiation of t(8;21) cells by elevated expression of ADA and downregulation of JUN. RUNX1‑RUNX1T1 and MAPK1 depletion induced cell cycle arrest at the G0/G1 phase. Accumulation of cells in the G1 phase was largely the result of downregulated expression of TBRG4, CCNE2, FOXO4, CDK6, ING4, IL8, MAD2L1 and CCNG2 in the case of RUNX1‑RUNX1T1 depletion and increased expression of RASSF1, FBXO6, DADD45A and P53 in the case of MAPK1 depletion. Taken together, the current results demonstrate that MAPK1 promotes myeloid cell proliferation and differentiation simultaneously by cell cycle progression while suppresing apoptosis.


Quantification of Cholesterol Solubilized in Dietary Micelles: Dependence on Human Bile Salt Variability and the Presence of Dietary Food Ingredients.
The solubility of cholesterol in bile salt (BS) micelles is important to understand the availability of cholesterol for absorption in the intestinal epithelium and to develop strategies to decrease cholesterol intake from the intestinal lumen. This has been the subject of intense investigation, due to the established relation between the development of diseases such as atherosclerosis and high levels of cholesterol in the blood. In this work we quantify the effect of BS variability on the amount of cholesterol solubilized. The effect of some known hypocholesterolemic agents usually found in the diet is also evaluated, as well as some insight regarding the mechanisms involved. The results show that, depending on the bile salt composition, the average value of sterol per micelle is equal to or lower than 1. The amount of cholesterol solubilized in the BS micelles is essentially equal to its total concentration until the solubility limit is reached. Altogether, this indicates that the maximum cholesterol solubility in the BS micellar solution is the result of saturation of the aqueous phase and depends on the partition coefficient of cholesterol between the aqueous phase and the micellar pseudophase. The effect on cholesterol maximum solubility for several food ingredients usually encountered in the diet was characterized using methodology developed recently by us. This method allows the simultaneous quantification of both cholesterol and food ingredient solubilized in the BS micelles even in the presence of larger aggregates, therefore avoiding their physical separation with possible impacts on the overall equilibrium. The phytosterols stigmasterol and stigmastanol significantly decreased cholesterol solubility with a concomitant reduction in the total amount of sterol solubilized, most pronounced for stigmasterol. Those results point toward coprecipitation being the major cause for the decrease in cholesterol solubilization by the BS micelles. The presence of tocopherol and oleic acid leads to a small decrease in the amount of cholesterol solubilized while palmitic acid slightly increases the solubility of cholesterol. Those dietary food ingredients are completely solubilized by the BS micelles, indicating that the effects on cholesterol solubility are due to changes in the properties of the mixed micelles.


Dose uncertainty and resolution of polymer gel dosimetry using an MRI guided radiation therapy system's onboard 0.35 T scanner.
Magnetic Resonance Imaging (MRI) scanners are widely used for 3D gel dosimeters readout. However, limited access to MRI scanners is a challenge in MRI-based gel dosimetry. Recent clinical implementation of MRI-guided radiation therapy machines provides potential opportunities for onboard gel dosimetry using its MRI subsystem. The objective of this study was to investigate the feasibility of gel dosimetry using ViewRay's onboard 0.35 T MRI scanner. A BANG® polymer gel dosimeter was irradiated by three beams of 3 × 3 cm2 field size. The T2 relaxation rate (R2) of the irradiated gel was measured using a Philips 1.5 T Ingenia MRI and a ViewRay 0.35 T onboard MRI and spin-echo pulse sequences. The number of signal averages (NSA) was set to 16 for the ViewRay acquisitions and one for the Philips 1.5 T MRI to achieve similar signal-to-noise ratios. The in-plane spatial resolution was 1.5 × 1.5 mm2 and the slice thickness was 5 mm. The relative dose uncertainty was obtained using R2 versus dose curves to compare the performance of dosimetry using the two different MRIs and field strengths. The dose uncertainty decreased from 12% at 2 Gy to 3.5% at 7.5 Gy at 1.5 T. The dose uncertainty decreased from 13% at 2 Gy to 4% at 7.5 Gy with NSA = 16 and 3 × 3 mm2 pixel size, and from 10.5% at 2 Gy to 3.2% at 7.5 Gy with NSA = 16 and denoised R2 maps (1.5 × 1.5 mm2 pixel size) at 0.35 T. The mean of dose resolution was 0.4 Gy at 1.5 T while the mean of dose resolution was 0.8 Gy and 0.64 Gy at 0.35 T by downsampling and denoising the R2 map, respectively. Therefore, comparable dose uncertainty was achievable using the ViewRay's onboard 0.35 T and Philips 1.5 T MRI scanners. 3D gel dosimetry using onboard low-field MRI scanner provides ViewRay users a 3D high resolution dosimetry option besides film and ionization chamber.


Homologous Recombination Deficiency (HRD) Scoring, by Means of Two Different Shallow Whole-Genome Sequencing Pipelines (sWGS), in Ovarian Cancer Patients: A Comparison with Myriad MyChoice Assay.
High-grade serous ovarian cancer (HGSOC) patients carrying the BRCA1/2 mutation or deficient in the homologous recombination repair system (HRD) generally benefit from treatment with PARP inhibitors. Some international recommendations suggest that BRCA1/2 genetic testing should be offered for all newly diagnosed epithelial ovarian cancer, along with HRD assessment. Academic tests (ATs) are continuously under development, in order to break down the barriers patients encounter in accessing HRD testing. Two different methods for shallow whole-genome sequencing (sWGS) were compared to the reference assay, Myriad. All these three assays were performed on 20 retrospective HGSOC samples. Moreover, HRD results were correlated with the progression-free survival rate (PFS). Both sWGS chemistries showed good correlation with each other and a complete agreement, even when compared to the Myriad score. Our academic HRD assay categorized patients as HRD-Deficient, HRM-Mild and HRN-Negative. These three groups were matched with PFS, providing interesting findings in terms of HRD scoring and months of survival. Both our sWGS assays and the Myriad test correlated with the patient's response to treatments. Finally, our AT confirms its capability of determining HRD status, with the advantage of being faster, cheaper, and easier to carry out. Our results showed a prognostic value for the HRD score.


Ulnar digital compression neuropathy of the thumb caused by splinting.
Acute compression neuropathy of the ulnar digital nerve of the thumb is described in two women, 23 and 53 years of age. The injury was caused by pressure from a splint. The pathogenesis and clinical features resemble the classic "bowler's thumb." Early recognition and proper treatment will prevent irreversible neurologic damage.


Involvement of the mitogen-activated protein kinase SIMK in regulation of root hair tip growth.
Mitogen-activated protein kinases (MAPKs) are involved in stress signaling to the actin cytoskeleton in yeast and animals. We have analyzed the function of the stress-activated alfalfa MAP kinase SIMK in root hairs. In epidermal cells, SIMK is predominantly nuclear. During root hair formation, SIMK was activated and redistributed from the nucleus into growing tips of root hairs possessing dense F-actin meshworks. Actin depolymerization by latrunculin B resulted in SIMK relocation to the nucleus. Conversely, upon actin stabilization with jasplakinolide, SIMK co-localized with thick actin cables in the cytoplasm. Importantly, latrunculin B and jasplakinolide were both found to activate SIMK in a root-derived cell culture. Loss of tip-focused SIMK and actin was induced by the MAPK kinase inhibitor UO 126 and resulted in aberrant root hairs. UO 126 inhibited targeted vesicle trafficking and polarized growth of root hairs. In contrast, overexpression of gain-of-function SIMK induced rapid tip growth of root hairs and could bypass growth inhibition by UO 126. These data indicate that SIMK plays a crucial role in root hair tip growth.


Chimeric peptides: a new approach to enhancing the immunogenicity of peptides with low MHC class I affinity: application in antiviral vaccination.
Recruitment of the CTL repertoire specific for subdominant epitopes that have a low MHC class I-binding affinity could be the way to achieve an efficient protective immunity against spontaneous tumors and viruses with high mutation rate. However, we have reported recently that subdominant peptides of influenza A Puerto Rico/8/34 (flu PR8) nucleoprotein (NP) with low Db affinity are only partially able to protect mice against lethal influenza infection. This seems to be due to their inability to recruit the specific CTL repertoire, and suggests that subdominant peptides could be used for vaccination only if they become highly immunogenic. In this work, we describe an approach that allows enhancement of the immunogenicity of every low affinity peptide presented by the Db molecule. It consists in producing chimeric peptides composed by amino acids from a high Db affinity peptide (NP366) in positions that interact with the MHC, and amino acids from low Db affinity nonimmunogenic influenza NP-derived peptides (NP17, NP97, NP330, and NP469) in positions that are exposed to the TCR. All chimeric peptides tested exhibited a high Db affinity and efficiently recruited the CTL repertoire specific for the corresponding low Db affinity peptide. Furthermore, vaccination with chimeric peptides that corresponded to subdominant NP17 and NP97 peptides induced a very potent anti-flu PR8 protective immunity.


Serological and molecular prevalence of Borrelia burgdorferi, Anaplasma phagocytophilum, and Ehrlichia species in dogs from Minnesota.
A population of 731 naturally exposed pet dogs examined at a private practice in Baxter, Minnesota, an area endemic for Lyme disease and anaplasmosis, was tested by serological and molecular methods for evidence of exposure to or infection with selected vector-borne pathogens. Serum samples were tested by enzyme-linked immunosorbent assay (ELISA) for Anaplasma phagocytophilum, Borrelia burgdorferi, and Ehrlichia canis antibodies and for Dirofilaria immitis antigen. Blood samples from 273 dogs were also analyzed by polymerase chain reaction (PCR) for Anaplasma and Ehrlichia species DNA. Based on the owner history and the attending veterinarian's physical examination findings, dogs exhibiting illness compatible with anaplasmosis or borreliosis were considered clinical cases, and their results were compared to the healthy dog population. Antibodies to only A. phagocytophilum were detected in 217 (29%) dogs; to only B. burgdorferi, in 80 (11%) dogs; and seroreactivity to both organisms, in 188 (25%) dogs. Of 89 suspected cases of canine anaplasmosis or borreliosis, A. phagocytophilum or B. burgdorferi antibodies were detected in 22 dogs (25%) and 8 dogs (9%) respectively, whereas antibodies to both organisms were found in 38 dogs (43%). Ehrlichia canis antibodies and D. immitis antigen were each detected in 11 (1.5%) dogs. Anaplasma phagocytophilum DNA was amplified from 7 of 222 (3%) healthy dogs and 19 of 51 (37%) clinical cases. Seroreactivity to both A. phagocytophilum and B. burgdorferi was detected more frequently in suspected cases of anaplasmosis and/or borreliosis than seroreactivity to either organism alone. Based on PCR testing, A. phagocytophilum DNA was more prevalent in suspected cases of anaplasmosis or borreliosis than in healthy dogs from the same region.


Estimating Direct and Indirect Protective Effect of Influenza Vaccination in the United States.
With influenza vaccination rates in the United States recently exceeding 45% of the population, it is important to understand the impact that vaccination is having on influenza transmission. In this study, we used a Bayesian modeling approach, combined with a simple dynamical model of influenza transmission, to estimate this impact. The combined framework synthesized evidence from a range of data sources relating to influenza transmission and vaccination in the United States. We found that, for seasonal epidemics, the number of infections averted ranged from 9.6 million in the 2006-2007 season (95% credible interval (CI): 8.7, 10.9) to 37.2 million (95% CI: 34.1, 39.6) in the 2012-2013 season. Expressed in relative terms, the proportion averted ranged from 20.8% (95% CI: 16.8, 24.3) of potential infections in the 2011-2012 season to 47.5% (95% CI: 43.7, 50.8) in the 2008-2009 season. The percentage averted was only 1.04% (95% CI: 0.15, 3.2) for the 2009 H1N1 pandemic, owing to the late timing of the vaccination program in relation to the pandemic in the Northern hemisphere. In the future, further vaccination coverage, as well as improved influenza vaccines (especially those offering better protection in the elderly), could have an even stronger effect on annual influenza epidemics.


Hypothalamus-related resting brain network underlying short-term acupuncture treatment in primary hypertension.
The present study attempted to explore modulated hypothalamus-seeded resting brain network underlying the cardiovascular system in primary hypertensive patients after short-term acupuncture treatment. Thirty right-handed patients (14 male) were divided randomly into acupuncture and control groups. The acupuncture group received a continuous five-day acupuncture treatment and undertook three resting-state fMRI scans and 24-hour ambulatory blood pressure monitoring (ABPM) as well as SF-36 questionnaires before, after, and one month after acupuncture treatment. The control group undertook fMRI scans and 24-hour ABPM. For verum acupuncture, average blood pressure (BP) and heart rate (HR) decreased after treatment but showed no statistical differences. There were no significant differences in BP and HR between the acupuncture and control groups. Notably, SF-36 indicated that bodily pain (P = 0.005) decreased and vitality (P = 0.036) increased after acupuncture compared to the baseline. The hypothalamus-related brain network showed increased functional connectivity with the medulla, brainstem, cerebellum, limbic system, thalamus, and frontal lobes. In conclusion, short-term acupuncture did not decrease BP significantly but appeared to improve body pain and vitality. Acupuncture may regulate the cardiovascular system through a complicated brain network from the cortical level, the hypothalamus, and the brainstem.


[Upper limb work-related musculoskeletal disorders (UL-WMSDs): a retrospective cohort study in three large factories of the upholstered furniture industry].
The epidemiological evidence of work-related musculoskeletal disorders (UL-WMSDs) due to repetitive strain and movements in the various industries has been collected in the literature mainly through cross-sectional surveys. In particular there are no contributions so far regarding the upholstered furniture industry with a longitudinal design. The aim of the study was to evaluate the incidence rate of WMSDs such as hand-wrist and shoulder tendonitis, carpal tunnel syndrome, and epicondylitis in exposed workers of three large companies of the upholstered furniture industry in a large geographic area of southern Italy. The OCRA method, recommended by international standard ISO 11228-3 and EN 1005-5, was used for risk assessment. The following work tasks were considered:.filling preparation workers, leather-cutting operators, sewing and upholstery-assembly workers. A total population of 5,278 subjects (exposed n=2927, controls n=2351) was investigated. The person/year at risk parameters were calculated from 1 January 2000, or from the date of engagement if later, until the first diagnosis of WMSD or, in absence of disorders, until the end of the study, i.e. 31 December 2004. Disorders occurring after the first were not considered. A multiple regression analysis was used to evaluate relationships between rates. Incidence rates correlated with risk classes of the OCRA index. An incidence rate of WMSDs higher than 1.2 cases per 100 person/year may be considered as a threshold value to suspect an occupational exposure to repetitive strain and movements warranting further investigation. The analysis of single factors did not show a greater predisposition of the female gender, with the single exception of the carpal tunnel syndrome (RR 2.92; 95% CI 1.57-5.43). Shoulder disorders affected mainly male leather-cutting operators (RR 4.97; 95% CI 2.03-12.16) and among all the factors influencing risk (frequency, force, posture, additional risk factors, pauses) posture seems to play a significant role.


Trends in mortality and morbidity for very low birth weight infants, 1991-1999.
Medical care for very low birth weight (VLBW) infants and their mothers has changed dramatically during the 1990s, yet it is unclear how these changes have affected mortality and morbidity. We used the Vermont Oxford Network Database to identify trends in clinical practice and patient outcomes for VLBW infants born from 1991 to 1999. Logistic regression was used to evaluate temporal trends in practices and outcomes while adjusting for patient characteristics and accounting for clustering of cases within hospitals. There were 118 448 infants 501 to 1500 g from 362 neonatal intensive care units enrolled in the Network Database from 1991 to 1999. Prenatal care, cesarean section, multiple births, antenatal steroids, and 1-minute Apgar scores increased during this period, as did the use of nasal continuous positive airway pressure, high-frequency ventilation, surfactant, and postnatal steroids. The proportion of white infants decreased; the proportions of Hispanic infants and those of other races increased. The crude and adjusted rates of mortality, pneumothorax, intraventricular hemorrhage (IVH), and severe IVH declined from 1991 to 1995, whereas from 1995 to 1999, the rates of mortality, IVH, and severe IVH did not change significantly, and pneumothorax increased. There have been major changes in both obstetric and neonatal care during the 1990s. These changes were associated with decreases in mortality and morbidity for VLBW infants during the first half of the decade. However, since 1995, no additional improvements in mortality or morbidity have been seen, ending a decades-long trend of improving outcomes for these infants.


Inframanubrial parathyroid glands in patients with primary hyperparathyroidism: alternatives to sternotomy.
Deep inframanubrial parathyroid tumors have traditionally been excised through a median sternotomy. With the advent of minimally invasive surgical access, we chose to examine the treatment options and outcomes of patients with inframanubrial mediastinal parathyroid tumors. Patients with primary hyperparathyroidism seen at a university medical center over a 12-year period were retrospectively reviewed. The utility of localization studies, methods of treatment, complications, and outcomes were examined in patients with a parathyroid tumor located in the mediastinum inferior to the manubrium. Patients with parathyroid adenomas located at the thoracic inlet were excluded. Sixteen patients with inframanubrial mediastinal tumors were treated during the study period. Altogether, 81% of the patients had undergone at least one prior neck exploration for primary hyperparathyroidism. Preoperative calcium and parathyroid hormone levels were 12.4 +/- 0.36 mg/dl and 273 +/- 70 pg/ml, respectively. Localization studies identified mediastinal parathyroid adenomas in the following locations: anterior mediastinum (n = 8), middle mediastinum (n = 7), posterior mediastinum (n = 1). Mediastinal computed tomography and technetium-sestamibi scans demonstrated the best sensitivity, 92% and 85%, respectively. Seven patients underwent successful excision of the mediastinal adenoma by transcervical mediastinal exploration with the Cooper retractor. The other patients underwent angiographic ablation (n = 4), anterior mediastinotomy (n = 3), video-assisted thoracoscopy (VATS) (n = 1), and VATS plus thoracotomy (n = 1). The mean hospital stay for the study group was 2.9 +/- 0.7 days. The complication rate was 25%. All patients were normocalcemic after a mean follow-up of 15 +/- 7 months. Most inframanubrial mediastinal parathyroid tumors can be successfully managed without median sternotomy.


Effects of therapeutic touch on the vital signs of patients before coronary artery bypass graft surgery.
Currently healthy heart word considered to be the objective of community health applications in many countries of the world because cardiovascular diseases are the most important factor in mortality of humans, worldwide. Coronary artery bypass graft surgery is one of the most common surgery procedures for these patients. The purpose of this study is to assess the impact of therapeutic touch on medical vital signs of patients before coronary artery bypass graft surgery. The present study is a clinical trial with 44 samples that were selected by easy sampling method and based on two separate lists of random numbers for both men and women; they were divided into two groups. In the therapeutic touch group, intervention therapy was applied on patents for 20 minutes. Data was analyzed using descriptive and inferential statistics. Test results showed that there was a significant difference between the mean pulse rate before and after intervention in both groups (p < 0.001). Results also showed that there was a significant difference between the average number of breathing before and after intervention in both groups (p < 0.001). Considering the effects of therapeutic touch therapy as a safe and effective intervention on the patients which were revealed in this study, this technique can be used as a simple, cheap and applicable technique in all health care centers to help these patients.


Natural mutations in the receptor binding domain of spike glycoprotein determine the reactivity of cross-neutralization between palm civet coronavirus and severe acute respiratory syndrome coronavirus.
The severe acute respiratory syndrome (SARS) outbreak of 2002 and 2003 occurred as a result of zoonotic transmission. Coronavirus (CoV) found in naturally infected palm civet (civet-CoV) represents the closest genetic relative to SARS-CoV, but the degree and the determinants of cross-neutralization among these viruses remain to be investigated. Studies indicate that the receptor binding domain (RBD) of the SARS-CoV spike (S) glycoprotein contains major determinants for viral entry and neutralization. We aim to characterize the impact of natural mutations within the RBDs of civet-CoVs on viral entry and cross-neutralization. In this study, the S glycoprotein genes were recovered from naturally infected civets in central China (Hubei province), extending the geographic distribution of civet-CoV beyond the southeastern province of Guangdong. Moreover, pseudoviruses generated in our laboratory with four civet S genes, each with a distinct RBD, infected cells expressing human receptor angiotensin-converting enzyme 2, but with 90 to 95% less efficiency compared to that of SARS-CoV. These four civet S genes were also constructed as DNA vaccines to immunize mice. Immunized sera elicited against most civet S glycoproteins displayed potent neutralizing activities against autologous viruses but were much less efficient (50% inhibitory concentration, 20- to 40-fold) at neutralizing SARS-CoV and vice versa. Convalescence-phase sera from humans were similarly ineffective against the dominant civet pseudovirus. Our findings suggest that the design of SARS vaccine should consider not only preventing the reemergence of SARS-CoV but also providing cross-protection, thus interrupting zoonotic transmission of a group of genetically divergent civet CoVs of broad geographic origin.


[Tryptophan loading and its plasma concentration in neurosurgical patients].
Tests for blood plasma tryptophan in patients with various diseases of the nervous system showed a statistically significant increase of its content in tumors of the astrocytic series in patients with meningiomas and oligodendrogliomas. The character of the tryptophan excretion curve after a load in neurooncological patients was different from that in healthy individuals and patients with thrombosis of the sinus and arachnoencephalitis. Hydrocortisone injection normalized the tryptophan curve in patients and, consequently, also the tryptophanpyrrolase reaction which is evidently disturbed in neurooncological patients.


Phytochemical analysis of Pinus eldarica bark.
Bark extract of Pinus pinaster contains numerous phenolic compounds such as catechins, taxifolin, and phenolic acids. These compounds have received considerable attentions because of their anti-inflammatory, antimutagenic, anticarcinogenic, antimetastatic and high antioxidant activities. Although P. pinaster bark has been intensely investigated in the past; there is comparably less information available in the literature in regard to P. eldarica bark. Therefore, the aim of this study was to determine the chemical composition of P. eldarica commonly found in Iran. A reversed-phase high pressure liquid chromatography (RP-HPLC) method for the determination of catechin, caffeic acid, ferulic acid, and taxifolin in P. pinaster and P. eldarica was developed. A mixture of 0.1% formic acid in deionized water and 0.1% formic acid in acetonitrile was used as the mobile phase, and chromatographic separation was achieved on a Nova pack C18 at 280 nm. The two studied Pinus species contained high amounts of polyphenolic compounds. Among four marker compounds, the main substances identified in P. pinaster and P. eldarica were taxifolin and catechin, respectively. Furthermore, the composition of the bark oil of P. eldarica obtained by hydrodistillation was analyzed by gas chromatography/mass spectroscopy (GC/MS). Thirty-three compounds accounting for 95.1 % of the oil were identified. The oils consisted mainly of mono- and sesquiterpenoid fractions, especially α-pinene (24.6%), caryophyllene oxide (14.0%), δ-3-carene (10.7%), (E)-β-caryophyllene (7.9%), and myrtenal (3.1%).


Mobilizing the research ecosystem for scientific advances towards positive impact in the context of the COVID-19 Pandemic
This special issue of the Turkish Journal of Medical Sciences is dedicated to providing scientific advances in the process of better understanding the SARS-CoV-2 virus that causes the COVID-19 infection. The special issue is published in a special time in which science-based approaches, cocreation-based collaboration, and the effective utilization and integration of competences have a crucial role during the race against time while combating the COVID-19 pandemic. In this process, the Scientific and Technological Research Council of Turkey (TÜBİTAK), which publishes academic journals including the Turkish Journal of Medical Sciences, has taken rapid action to mobilize the research community. This includes forming new scientific coalitions in record time, the opening of new calls across the research ecosystem, the organization of a virtual scientific conference, and the launch of a new portal in support of cocreation processes and open science. In addition, various teleconferences that bring together various disciplines at the national and international level have taken place. All of these efforts provide multiple venues to support the common effort of combating the COVID-19 pandemic with R&D and development as a common objective. The sharing of evidence-based knowledge and scientific progress is an effective approach towards providing important contributions for combating the COVID-19 pandemic. The research articles that are contained in this special issue of the Turkish Journal of Medical Sciences involves a special collection dedicated to COVID-19. This short communication aims to provide an introduction of the major initiatives that have been taken in the scientific landscape with a focus on Turkey.


Size effects and stochastic behavior of nanoindentation pop in.
A statistical model for pop in initiated at preexisting dislocations during nanoindentation is developed to explain size-dependent pop-in stresses. To verify theoretical predictions of this model, experiments were performed on single-crystal Mo, utilizing indenter radii that vary by over 3 orders of magnitude. The stress where plastic deformation begins ranges from the theoretical strength in small volumes, to 1 order of magnitude lower in larger volumes. An intermediate regime shows wide variability in the stress to initiate plastic deformation. Our theory accurately reproduces the experimental cumulative probability distributions, and predicts a scaling behavior that matches experimental behavior.


Characterization of a murine model of recurrent herpes simplex viral keratitis induced by ultraviolet B radiation.
The authors characterized a murine model of herpes simplex virus (HSV) reactivation in which recurrent herpetic keratitis was obtained in up to 80% of animals. Five weeks after ganglionic latency was established in National Institutes of Health inbred mice after corneal inoculation, HSV type 1 (HSV-1) was reactivated by irradiating the previously inoculated eye with ultraviolet (UV) light. Comparison of different UV wavelengths showed UVB to be optimal for reactivation, with peak viral recurrence being induced by a total exposure of approximately 250 mJ/cm2. Reactivated infectious virus generally began to appear in trigeminal ganglia 2 days postirradiation and was subsequently detectable in the cornea by both corneal swabbing and immunostaining for viral antigens. Two consecutive outbreaks of viral recurrence at the ocular surface were induced in selected animals by serial exposure to UVB. Advantages of this model over other models of recurrent keratitis are discussed.


[Antiamebic effect of metronidazole proved in a study conducted in Cienfuegos province].
Three studies were conducted in Cienfuegos province to prove that intestinal amebiasis in Cuba could be an overestimated health problem. The first two studies showed two components of this overestimation: microscopical overdiagnosis and lack of knowledge about Entamoeba dispar, a non-pathogenic species, in most cases in which microscopical examination was correct. This paper reported the third study that showed the third component: the wrong belief that Entamoeba histolytica is resistant to metronidazole. Thirty-five individuals infected with one or both species of E-histolytica-E. dispar combination were given metronidazole at a dose of 250 mg three times daily for 10 days. Stool samples taken immediately after the treatment were tested by ENZYMEBA to detect one or both species and by a multiplex polymerase chain reaction procedure to determine the existing species. The results of these assays revealed that E. histolytica infection disappeared in all the cases, therefore, we may conclude that in Cienfuegos province and probably in the rest of the country, metronidazol remains an effective drug in the treatment of intestinal amebiasis


IL28B polymorphism and cytomegalovirus predict response to treatment in Egyptian HCV type 4 patients.
To test whether the status of positive cytomegalovirus (CMV) DNA detection adds to the predictive value of IL28B and to further categorize C/T allele carriers. This study included 166 chronic hepatitis C (CHC) patients who received combined interferon and ribavirin therapy for 48 wk, 84 spontaneous hepatitis C virus (HCV) resolvers who were positive for IgG anti-HCV antibody and negative for HCV RNA, and 100 healthy subjects who were negative for both HCV antibodies and RNA as controls. Genomic DNA from peripheral blood was used for IL28B rs.12979860 single nucleotide polymorphism (SNP) and CMV DNA detection. A 139 bp fragment containing IL28B SNP was amplified in all subjects by polymerase chain reaction using a specifically designed primer. Then the IL28B rs.12979860 SNP was detected by restriction fragment length polymorphism (RFLP) genotyping. The presence of CMV DNA was tested by amplification of the gB1 gene using nested polymerase chain reaction. The role of CMV and IL28B rs.12979860 SNP genotypes in determining the response rate to combined interferon therapy and clinical status of patients were statistically analyzed. Current data showed that 67% of patients carrying the IL28B 12979860 C/C allele had a sustained viral response (SVR) while the genotypes C/T and TT were associated with lower SVR rates, 50% and 48%, respectively. SVR rates for the C/C allele were lower than other HCV genotypes and/or other populations. Genotype CC was associated with the response to interferon (P = 0.025). Genotype C/C was reduced from 48% in controls to 14% in CHC patients suggesting its protective role against progression to chronicity. The majority of spontaneously cleared subjects (86%) were C/C, confirming its protective role. The C/T allele was present in 71% of CHC patients compared with 38% of controls, so the use of IL28B SNP genotyping only in these patients may be of little value as a predictor of response. CMV reactivation occurred in 40% of CHC patients. Co-infection with CMV seriously diminished the response to interferon (IFN) therapy, with SVR rates in C/C genotypes 87.5% in CMV-negative patients and 12.5% in CMV-positive patients (P < 0.0001). SVR rates among C/T carriers were reduced to < 50% in patients with positive CMV DNA while the non-response rate doubled. These data indicate that a supplemental assay for CMV viremia adds to the prognostic value of IL28B genotyping. The results suggest that both genetic (i.e., spontaneous) and therapeutic (IFN-based therapy) arms are complementary in the battle against HCV. CMV DNA testing may be of value to better predict the response to IFN, particularly in IL28B C/T carriers.


Synthesis and Characterization of Quaternized Poly(β-amino ester) for Highly Efficient Delivery of Small Interfering RNA.
The development of non-viral vectors for gene delivery has gained attention over the past decades. Specifically, poly(β-amino ester) (PBAE) has shown great potential for improving the delivery of gene therapeutics. It has been observed that low-molecular-weight PBAE displayed low transfection activities, while quaternization could enhance the transgene expression efficacy of PBAE. Herein, PBAE quaternary ammonium salt (PBAEQAS) was synthesized to increase the positive charge of the polymers, which resulted in an increase in siRNA binding efficiency based on self-assembly electrostatic interaction. Specifically, the nanoparticle surface was positively charged, which increased the uptake ability of siRNA. Compared with acrylate-PBAEQAS/siNC nanoparticles and amine-PBAEQAS/siNC nanoparticles, acrylate-PBAEQAS/siSurvivin nanoparticles and amine-PBAEQAS/siSurvivin nanoparticles induced more-efficient cell apoptosis and gene silencing. All these results suggest that PBAEQAS would be a promising gene delivery vector for cancer treatment.


Sustainable Approach for Recovery of Sulfur from Phophogypsum.
Phosphogypsum releases have a detrimental effect on flora and fauna and also cause economic loss due to the loss of sulfur. This study contributes to solving this problem using a process that consists of the valorization of this byproduct. A new sustainable method that consists of using sulfuric acid is proposed to process the Algerian phosphogypsum to recover the sulfur, which is imported in huge quantities until today. The experiments carried out in this investigation have enabled us to recover sulfur in the form of SO2 using appropriate additives such as silica, alumina, clay, and charcoal as reducing agents. These additives accelerate the desulfurization process and decrease the decomposition temperature of calcium sulfate. As a result, these additives allow the reduction of energy used and simultaneously increase the concentration of SO2. X-ray diffraction shows that thermochemical decomposition is not complete and that the sulfur present in the residue exists in the form of CaSO4 and CaS at 1150 °C. After calcination of phosphogypsum, the analysis of the residues obtained shows that they have a chemical composition almost identical in terms of quality to that of clinker, which can further be used in the cement production.


Microsurgical repair of the adolescent varicocele.
Since clinically apparent varicoceles may affect testicular volume and sperm production, early repair has been advocated. However, repair of the pediatric varicocele with conventional nonmagnified techniques may result in persistence of the varicocele after up to 16% of these procedures. Also testicular artery injury and postoperative hydrocele formation can occur after nonmagnified repair. The microsurgical technique has been successfully completed in a large series of adults with a dramatic reduction in complication and recurrence rates. We report our experience with the microsurgical technique in boys. A total of 30 boys (average age 15.9 years) underwent 42 microsurgical varicocelectomies (12 bilateral). All patients had a large left varicocele. Indications for repair included testicular atrophy (size difference between testicles of greater than 2 ml.) in 20 boys, pain in 5 and a large varicocele without pain or testicular atrophy in 5. Six boys were referred following failure of conventional nonmicrosurgical techniques. All boys were examined no sooner than 1 month postoperatively (mean followup 12). Preoperative volume of the affected testis averaged 13.0 ml., and an average size discrepancy between testicles of 2.8 ml. was noted before unilateral varicocelectomy. No cases of persistent or recurrent varicoceles were detected, and 1 postoperative hydrocele resolved spontaneously. After unilateral varicocelectomy the treated testes grew an average of 50.1%, while the contralateral testes grew only 23%. Overall, 89% of patients with testicular atrophy demonstrated reversal of testicular growth retardation after unilateral varicocelectomy. In contrast, both testes showed similar growth rates after bilateral varicocelectomy (45% left testis, 39% right testis). The meticulous dissection necessary to preserve arterial and lymphatic supply, and to ligate all spermatic veins in the pediatric patient is readily accomplished using a microsurgical approach, and results in low recurrence and complication rates. Rapid catch-up growth of the affected testis after microsurgical varicocelectomy suggests that intervention during adolescence is effective and warranted.


Treatment of T-lineage acute lymphoblastic leukemia.
T-cell acute lymphoblastic leukemia is an aggressive disease that responds poorly to "standard" therapy designed for the more common B-lineage ALLs in childhood. The principles of this "standard" therapy were derived from empiric clinical trials. Thus, it is not surprising that the therapy that had the greatest impact on survival in the group as a whole would be found to be most successful for the most common subset of patients. T-cell malignant lymphoblasts share many biologic features that set them apart from the more common B-lineage lymphoblasts. Some of these biologic features suggest therapeutic approaches that should be particularly successful in treating patients with T-cell leukemia. The use of aggressive, multiple-agent "pulse" chemotherapy has been shown through empiric trials to have relative efficacy in T-cell lymphoblastic leukemia, presumably because of the rapid generation time and high growth fraction. Future studies will (1) determine the optimal dose and schedule of cytosine arabinoside needed to exploit the increased Ara-CTP accumulation in T-cell blasts, (2) determine the efficacy of a new agent, deoxycoformycin, an inhibitor of adenosine deaminase, to exploit the biochemical phenotype of T-cell blasts, and (3) assess the ability of conjugated anti-T monoclonal antibodies to deliver a cytotoxic agent, thus exploiting unique antigenic determinants at the cell surface. As more is learned about the biology of T-cell malignancies, further treatment strategies may be suggested to exploit the new features that are discovered. Similarly, it is hoped that the unique features of the B-lineage leukemias will suggest treatment strategies that will improve survival in those patients as well. Certainly, improved survival has already been achieved in the case of the B-cell leukemias and Burkitt's lymphomas, and improvement may also be possible for the pre-B and early pre-B phenotypes of lymphoblastic leukemia.


Auditory N2 Correlates of Treatment Response in Posttraumatic Stress Disorder.
Emotional processing and cognitive control are implicated as being dysfunctional in posttraumatic stress disorder (PTSD) and targeted in cognitive processing therapy (CPT), a trauma-focused treatment for PTSD. The N2 event-related potential has been interpreted in the context of emotional processing and cognitive control. In this analysis of secondary outcome measures from a randomized controlled trial, we investigated the latency and amplitude changes of the N2 in responses to task-relevant target tones and task-irrelevant distractor sounds (e.g., a trauma-related gunshot and a trauma-unrelated lion's roar) and the associations between these responses and PTSD symptom changes. United States military veterans (N = 60) diagnosed with combat-related PTSD were randomized to either active or sham repetitive transcranial magnetic stimulation (rTMS) and received a CPT intervention that included a written trauma account element (CPT+A). Participants were tested before and 6 months after protocol completion. Reduction in N2 amplitude to the gunshot stimulus was correlated with reductions in reexperiencing, |r| = .445, and hyperarousal measures, |r| = .364. In addition, in both groups, the latency of the N2 event-related potential to the distractors became longer with treatment and the N2 latency to the task-relevant stimulus became shorter, ηp 2 = .064, both of which are consistent with improved cognitive control. There were no between-group differences in N2 amplitude and latency. Normalized N2 latencies, reduced N2 amplitude to threatening distractors, and the correlation between N2 amplitude reduction and PTSD symptom reduction reflect improved cognitive control, consistent with the CPT+A objective of addressing patients' abilities to respond more appropriately to trauma triggers.


A Medley Correlation of Serum Zinc with Glycemic Parameters in T2DM Patients.
Studies in diabetic have reported zinc deficiency due to zincuria. Effective treatment of oral antidiabetic drugs should improve glycemic status correcting serum zinc levels. This study evaluated serum zinc level and its correlation with glycemic parameters in type 2 Diabetes Mellitus (T2DM) patients receiving oral antidiabetics. To study correlation of serum zinc and glycemic parameters in patients receiving oral antidiabetics. It was a prospective cross sectional pilot study, conducted for 1 year, with the approval of IEC. Patients of T2DM satisfying inclusion/exclusion criteria were enrolled. Serum zinc and glycemic parameters were estimated. The data was stratified into- Group A: Metformin (n = 20), Group B: Metformin and glimiperide (n = 13). Correlation analysis of serum zinc and glycemic parameters was carried. The mean age and duration of 33 patients was 57 ± 9.1 and 6.30 ± 6.52 years respectively. The mean FBG, PPBG, HbA1c and zinc were 164 ± 35, 257 ± 63 mg/dL, 9.3 ± 2.2% and 58 ± 23 ug/dL respectively. Thirty patients had HbA1c >6.5%. The percentage of zinc deficiency was 76.92 and 90 in group A and B, respectively. Correlation of serum zinc and glycemic parameters was insignificant in overall group. It varied at different HbA1c levels and in different groups. A positive correlation existed between serum zinc level and HbA1c at ≥9.5%. Zinc deficiency was common in T2DM and to a greater extent in combination group. Correlation of serum zinc levels with glycemic parameters varied at different HbA1c and treatment groups.


Comparative study of two different rat models of stress urinary incontinence.
The purpose of this study was to compare the success rate and urodynamic characteristics of the rat model of stress Urinary incontinence established by ovariectomy at different times after vaginal dilation. Eighty female rats were randomly divided into three groups: the control group (n = 20), model group A (n = 30), i.e., the animals received colpectasis and bilateral oophorectomy within 1 day; model group B (n = 30), i.e., the animals received bilateral oophorectomy 1 week after colpectasis. The sneeze test and urodynamics were tested. There was no significant difference between the model group A and the model group B (p > 0.05) in the sneeze test. The leak point pressure (LPP) of model groups A and B was lower than that of the control group (p < 0.05). The LPP value of model group A was lower than that of model group B (p < 0.05). There was no significant difference in maximum bladder capacity (MBC) between model group A and the control group (p > 0.05); the MBC value of model group B was lower than that of the control group and model group A (p < 0.05). Models of stress urinary incontinence in rats can be successfully established by both modeling methods. In terms of LPP, model groups A and B had lower values than the control group, and model group A had lower values than model group B. In terms of MBC, there was no difference between model group A and the control group, and model group B had lower values than the control group and model group A.


Complications of pelvic exenteration.
This report is based on a retrospective review of 104 patients who had undergone pelvic exenteration for advanced malignancy over a 29-year period (1956 to 1984, inclusive). Fifty-one patients (49%) developed major complications of the operative field involving the gastrointestinal tract (fistula or obstruction), the urinary tract (fistula, infection, or obstruction), or the wound (abscess, dehiscence/necrosis, or hemorrhage). No association was identified between the complication rate and organ of primary disease, extent of disease, tumor histology, or extent of resection. Patients receiving pelvic radiotherapy prior to exenteration had a much higher complication rate (39/58, 67%) than patients having had no radiotherapy (12/46, 26%). Reconstruction of the irradiated pelvis after exenteration by omental flap, colonic advancement, and/or myocutaneous flaps decreased the complication rate from 82% (27/33) to 48% (12/25). The operative mortality of pelvic exenteration was 2.9% and the actuarial five-year survival rate was 27%.


A review of 2,413 tumours operated over a 30-year period.
Studying 2,413 cases of supratentorial brain tumors operated on a 30 years period, the author emphasizes appealing signs, pitfalls, and tumoral relapses. Epilepsy is the most frequent initial sign preceding headache and focal signs. We discuss the frequency of the epilepsy, the age of the first seizure, the location and the histological nature of the epiletogenic tumor, and the type of the seizures. In the end, we study diagnosis pitfalls such as long-standing epilepsy, ictal presentation, pitfalls due to metastases and the difficult problem of tumoral relapses.


[Current studies useful for indications in the treatment of varicose veins of the lower limbs. Preoperative explorations].
The preoperative assessment of varicose veins usually requires the use of ultrasound methods to specify and localize the various sources of reflux in the saphenous junctions, perforating veins and saphenous branches. Continuous wave Doppler (CW) is the basic technique used to investigate reflux and usefully completes clinical examination of varicose veins. Duplex-scan provides morphological analysis of the veins and studies the direction of flow in a precise site. It is particularly useful to study the saphenous-junction, perforating veins and deep vems. Color Doppler imaging (CDI) offers the advantage of duplex-scanning, withe visualization of great volumes of measurement whit blue or red colour-coding according to the direction of flow. All haemodynamic and morphological findings must be reported on a diagram to establish "mapping" of pathological veins.


Improving Care by Decreasing Wait Times: A Military Health System Quality Improvement Project.
Maximizing patient satisfaction is now a significant focus in many outpatient orthopedic clinics as it has been shown to affect many facets of healthcare. With this quality improvement study, we sought to determine the areas of the longest patient wait time during their clinical encounter and evaluate the effect of an identification card system on those times. We hypothesized that utilizing an identification card system would expedite the time patients spend in the clinic and decrease wait times. This quality improvement study was conducted in an orthopedic hand surgery clinic within a military treatment facility. Pre-intervention and intervention data collection consisted of a time sheet that was filled out by surgeons and clinic staff as the patient progressed through their appointment. The intervention consisted of a card system to track the patient throughout their encounter and mark their order in the queue. There were a total of 130 patients in the pre-intervention group and 113 in the intervention group. Compared with the pre-intervention group, the intervention group patients had significantly less time from check-in to being roomed by the clinic staff with a mean of 13 ± 13 vs. 21 ± 19 minutes in the pre-intervention group (P < .001). In the pre-intervention cohort, the average clinic encounter was 66 ± 38 minutes compared to 57 ± 35 minutes in the intervention cohort, which was not statistically significant (P = .112). The time spent with the orthopedic surgeon and occupational therapists increased in the intervention group. We provided a simple way to decrease patient wait times and increase time with the healthcare team, utilizing our existing clinic space. Time with the orthopedic surgeon significantly increased because of our intervention, while the overall clinic time trended down.


Amino acid residues of G-protein-coupled receptors critical for endoplasmic reticulum export and trafficking.
Analysis of the structural features of rhodopsin-type G-protein-coupled receptors (GPCRs) revealed the existence of an additional α-helix, termed helix 8, in the C-terminal tail. Furthermore, these GPCRs were determined to possess several conserved residues in their transmembrane domains. The functional deficiencies of receptors in which these domains or residues have been mutated have not been examined in living cells due to their accumulation in the endoplasmic reticulum (ER), although the ligand affinities of these receptors have been tested in membrane preparations. Recent studies have demonstrated that ER-accumulated receptors are effectively exported from ER using membrane permeable ligands as pharmacological chaperones. Here, we identified several residues of the platelet-activating factor receptor and leukotriene B(4) type-II receptor that are crucial for export from ER. Moreover, we used their specific ligands as pharmacological chaperones to traffic ER-accumulated GPCRs to the cell surface in order to examine the functional deficiencies of each mutant receptor. Here, we introduce the novel technique of site-specific N-terminal labeling of cell surface proteins in living cells with Sortase-A, a transpeptidase isolated from Staphylococcus aureus, to evaluate the trafficking of receptors after agonist stimulation.


Making decisions for incapable adults 4: participation in research.
In the final article on the Mental Capacity Act 2005, Richard Griffith discusses the safeguards in place to protect incapable persons who may be the subject of a research study or clinical trial. District nurses and other health professionals who wish to conduct research involving people with incapacity will have a duty to apply the 2005 Act's safeguards to ensure that the research is lawful.


IFN-gamma and CD8+ T cells restore host defenses against Pneumocystis carinii in mice depleted of CD4+ T cells.
Host defenses against infection are profoundly compromised in HIV-infected hosts due to progressive depletion of CD4+ T lymphocytes and defective cell-mediated immunity. Although recent advances in antiretroviral therapy can dramatically lower HIV viral load, blood CD4+ T lymphocytes are not restored to normal levels. Therefore, we investigated mechanisms of host defense other than those involving CD4+ T lymphocytes against a common HIV-related opportunistic infection, Pneumocystis carinii (PC) pneumonia. Using CD4-depleted mice, which are permissive for chronic PC infection, we show that up-regulation of murine IFN-gamma by gene transfer into the lung tissue results in clearance of PC from the lungs in the absence of CD4+ lymphocytes. This resolution of infection was associated with a >4-fold increase in recruited CD8+ T lymphocytes and NK cells into the lungs. The role of CD8+ T cells as effector cells in this model was further confirmed by a lack of an effect of IFN-gamma gene transfer in scid mice or mice depleted of both CD4+ and CD8+ T cells. Cytokine mRNA analysis revealed that recruited, lung-derived CD8+ T cells had greater expression of IFN-gamma message in animals treated with the IFN-gamma gene. These results indicate that CD8+ T cells are capable of clearing PC pneumonia in the absence of CD4+ T cells and that this host defense function of CD8+ T cells, as well as their cytokine repertoire, can be up-regulated through cytokine gene transfer.


The Upf1 protein restricts EV-A71 viral replication.
Enterovirus A71 (EV-A71) is transmitted through the respiratory tract, gastrointestinal system, and fecal-oral routes. The main symptoms caused by EV-A71 are hand, foot, and mouth disease (HFMD) or vesicular sore throat. Upf1 (Up-frameshift protein 1) was reported to degrade mRNA containing early stop codons, known as nonsense-mediated decay (NMD). Upf1 is also involved in the NMD mechanism as a host factor detrimental to viral replication. In this study, we dissected the potential roles of Upf1 in the EV-A71-infected cells. Upf1 was virulently down-regulated in three different EV-A71-infected cells, RD, Hela, and 293T, implying that Upf1 is a host protein unfavorable for EV-A71 replication. Knockdown of Upf1 protein resulted in increased viral RNA expression and production of progeny virus, and conversely, overexpression of Upf1 protein resulted in decreased viral RNA expression and production of progeny virus. Importantly, we observed increased RNA levels of asparagine synthetase (ASNS), one of the indicator substrates for the NMD mechanism, which indirectly suggests that EV-A71 infection of cells suppresses NMD activity in the host. The results shown in this study are useful for subsequent analysis of the relationship between the NMD/Upf1 mechanism and other picornaviruses, which may lead to the development of anti-picornavirus drugs.


Effects of Dietary Garlic Extract on Growth, Feed Utilization and Whole Body Composition of Juvenile Sterlet Sturgeon (Acipenser ruthenus).
This study was carried out to investigate the supplemental effects of dietary garlic extract (GE) on growth performance of juvenile sterlet sturgeon (Acipenser ruthenus). The first experiment was designed to determine the optimum levels of garlic extract as growth promoter during 10 weeks. Three groups (two replicates/group) of 240 fish with mean body weight of 85 g were fed with diets containing 0 (control), 0.5 and 1.0% of GE. The highest weight gain (%) and feed efficiency (%) were found in fish groups fed with diet containing 0.5% GE. Subsequently, the supplemental effects of dietary GE was studied on growth of juvenile sterlet sturgeon (Acipenser ruthenus) with an average body weight of 59.6 g. Fish cultured in freshwater were randomly allotted to each of 10 tanks (two groups of five replicates, 20 fish/tank) and fed diets with 0.5% GE or without GE (control), respectively, at the level of 2.0% of fish body weight per day for 5 weeks. Weight gain (51.1%), feed efficiency (79.1%), specific growth rate (1.18%) and protein efficiency ratio (1.50) of fish fed 0.5% GE were significantly (p<0.05) higher than those fish fed the control diet. Significantly higher protein (PRE 20.4%) and lipid retention efficiencies (LRE, 74.5%) were also found in 0.5% GE group (p<0.05). The present results suggested that dietary GE could improve growth and feed utilization of juvenile sterlet sturgeons.


suPAR Cut-Offs for Risk Stratification in Patients With Symptoms of COVID-19.
Elevated soluble urokinase Plasminogen Activator Receptor (suPAR) is a biomarker associated with adverse outcomes. We aimed to investigate the associations between plasma suPAR levels (testing the cut-offs ⩽4, 4-6, and ⩾6 ng/mL) with risk of 14-day mortality, and with the risk of mechanical ventilation in patients that tested positive for SARS-CoV-2. Observational cohort study of patients presenting with symptoms of COVID-19 at Department of Emergency Medicine, Amager and Hvidovre Hospital, Denmark from March 19th, 2020 to April 3rd, 2020. Plasma suPAR was measured using suPARnostic technologies. Patients were followed for development of mechanical ventilation and mortality for 14 days. Validation of our findings were carried out in a similar sized COVID-19 patient cohort from Mikkeli Central Hospital, Finland. Among 386 patients with symptoms of COVID-19, the median (interquartile range) age was 64 years (46-77), 57% were women, median suPAR was 4.0 ng/mL (2.7-5.9). In total, 35 patients (9.1%) died during the 14 days follow-up. Patients with suPAR ⩽4 ng/mL (N = 196; 50.8%) had a low risk of mortality (N = 2; 1.0%; negative predictive value of 99.0%, specificity 55.3%, sensitivity 95.2%, positive predictive value 17.4%). Among patients with suPAR ⩾6 ng/mL (N = 92; 23.8%), 16 died (17.4%). About 99 patients (25.6%) tested positive for SARS CoV-2 and of those 12 (12.1%) developed need for mechanical ventilation. None of the SARS-CoV-2 positive patients with suPAR ⩽4 ng/mL (N = 28; 38.8%) needed mechanical ventilation or died. The Mikkeli Central Hospital validation cohort confirmed our findings concerning suPAR cut-offs for risk of development of mechanical ventilation and mortality. Patients with symptoms of COVID-19 and suPAR ⩽4 or ⩾6 ng/mL had low or high risk, respectively, concerning the need for mechanical ventilation or mortality. We suggest cut-offs for identification of risk groups in patients presenting to the ED with symptoms of or confirmed COVID-19.