Lessons learned from the hip and knee musculoskeletal outcomes data evaluation and management system.
In 1994 the American Academy of Orthopaedic Surgeons established an outcomes initiative. The Musculoskeletal Outcomes Data Evaluation and Management System had two goals: (1) to create validated patient-based functional health questionnaires that would become the gold standard for musculoskeletal research, and (2) to collect data from practicing orthopaedists using these instruments. We assessed the adequacy of the hip and knee portion of the Musculoskeletal Outcomes Data Evaluation and Management System data collection process to learn how to improve these processes. Database elements included demographic and clinical information and health and well-being scales. Only 715 records or 578 patients of the initial 2419 records (30%) had complete baseline and followup information for either hip or knee replacement procedure. Only 17% of the patients who had a knee replacement and 20% of the patients who had a hip replacement returned for followup between 7 and 9 months. The Musculoskeletal Outcomes Data Evaluation and Management System achieved its goal regarding the development of validated questionnaires, but the data collection component did not. The project's implementation and management were flawed. This experience offers the field of orthopaedics valuable lessons that can be applied to future large-scale data collection efforts.


Medicinal lavender modulates the enteric microbiota to protect against Citrobacter rodentium-induced colitis.
Inflammatory bowel disease, inclusive of Crohn's disease and ulcerative colitis, consists of immunologically mediated disorders involving the microbiota in the gastrointestinal tract. Lavender oil is a traditional medicine used to relieve many gastrointestinal disorders. The goal of this study was to examine the therapeutic effects of the essential oil obtained from a novel lavender cultivar, Lavandula×intermedia cultivar Okanagan lavender (OLEO), in a mouse model of acute colitis caused by Citrobacter rodentium. In colitic mice, oral gavage with OLEO resulted in less severe disease, including decreased morbidity and mortality, reduced intestinal tissue damage, and decreased infiltration of neutrophils and macrophages, with reduced levels of TNF-α, IFN-γ, IL-22, macrophage inflammatory protein-2α, and inducible nitric oxide synthase expression. This was associated with increased levels of regulatory T cell populations compared with untreated colitic mice. Recently, we demonstrated that the composition of the enteric microbiota affects susceptibility to C. rodentium-induced colitis. Here, we found that oral administration of OLEO induced microbiota enriched with members of the phylum Firmicutes, including segmented filamentous bacteria, which are known to protect against the damaging effects of C. rodentium. Additionally, during infection, OLEO treatment promoted the maintenance of microbiota loads, with specific increases in Firmicutes bacteria and decreases in γ-Proteobacteria. We observed that Firmicutes bacteria were intimately associated with the apical region of the intestinal epithelial cells during infection, suggesting that their protective effect was through contact with the gut wall. Finally, we show that OLEO inhibited C. rodentium growth and adherence to Caco-2 cells, primarily through the activities of 1,8-cineole and borneol. These results indicate that while OLEO promoted Firmicutes populations, it also controlled pathogen load through antimicrobial activity. Overall, our results reveal that OLEO can protect against colitis through the microbial-immunity nexus and that a pharmacological agent, in this case OLEO, alters the normal enteric microbiota.


Patients' preferences for new versus old anticoagulants: a mixed-method vignette-based study.
For both patients and clinicians, differences between older and new anticoagulants have major implications for treatment selection, day-to-day management of therapy and adherence. To explore patients' preferences for warfarin versus direct oral anticoagulant (DOAC) therapy. Mixed-method study involving anticoagulated older patients admitted to hospital. Part A comprised a vignette-based questionnaire; patients were asked whether they preferred Medicine A (warfarin) or Medicine B (DOAC). Part B interviews explored patients' satisfaction with their current anticoagulant. Responses were thematically analysed. Forty patients participated: 23 warfarin-treated, 17 DOAC-treated. Collectively, Parts A and B identified that most patients were satisfied with their current therapy (warfarin or DOAC), expressing reluctance to change to alternatives. Among patients who were able to numerically rate their satisfaction with therapy, most were 'satisfied' with their current anticoagulant, although warfarin-treated patients were slightly less 'satisfied' (median score 3.5) than those on DOACs (median score 5.0). Despite this, warfarin-treated patients still preferred their current therapy (over DOACs) due to familiarity and the security of regular international normalised ratio (INR) monitoring; those who preferred DOACs cited previous warfarin-related bleeding and unstable INRs as key reasons. DOAC-treated patients who preferred warfarin perceived regular monitoring as a major advantage; only those having had negative experiences with warfarin clearly preferred DOACs. Most patients accepted their currently prescribed anticoagulant, be it warfarin or DOACs. Features of specific anticoagulants, such as regular monitoring with warfarin, were perceived variably - some patients cited them as advantages and others as disadvantages. The clearest preference identified was for the agent already being taken.


Intercalation of tetracationic metalloporphyrins and related compounds into DNA.
Electron paramagnetic resonance (e.p.r.) studies of thin DNA-chelate films indicate that both the water soluble planar macrocyclic chelates, cobalt (II) tetra (2,3-N-methylpyridinium) porphyrazine (cobalt (II) TMTPPA), and vanadyl (II) 5,10,15,20-tetra-(4-methylpyridinium) porphyrin (Vo (II) TMTPyP) can bind to calf thymus DNA intercalatively. The intercalation of cobalt (II) porphyrazine is in contrast to the binding of cobalt (II) porphyrin, which is known to bind nonintercalatively. Five-coordinate cobalt (II) porphyrazine is sufficiently slim, even with its single out-of-plane ligand, to permit significant intercalative binding. Vanadyl (II) porphyrin intercalates into calf thymus DNA, but not as readily as cobalt (II) porphyrazine.


The actions of FMRFamide-like peptides on visceral and somatic muscles of the snail Helix aspersa.
Phe-Met-Arg-Phe-NH2 (FMRFamide) and pyroGlu-Asp-Pro-Phe-Leu-Arg-Phe-NH2 (pQDPFLRFamide) occur in the ganglia and tissues of the snail, Helix aspersa. This report describes the effects of these two neuropeptides on five visceral organs or somatic muscles isolated from the snail (Table 1). The epiphallus, as well as the rest of the male reproductive tract, was contracted by both FMRFamide and pQDPFLRFamide, and the threshold was usually below 5 X 10(-9) mol l-1 (Fig. 1). Both peptides also reduced the resting tone of the crop and decreased the force and frequency of its rhythmic activity; FMRFamide is about 10 times more potent (Fig. 4). In contrast, pQDPFLRFamide was about 100 times more potent than FMRFamide as a cardioexcitatory agent (Fig. 5). The actions of the peptides on the pharyngeal and tentacle retractor muscles were markedly different: FMRFamide primarily contracted these muscles; and pQDPFLRFamide usually had no effect alone, but relaxed or diminished contractions induced by FMRFamide and acetylcholine (ACh) (Figs 6, 8, 9). Other analogues of FMRFamide were tested, but none was as effective a relaxing agent as pQDPFLRFamide. The effects of FMRFamide and pQDPFLRFamide on all of the preparations could be distinguished from those produced by ACh and 5-hydroxytryptamine (5-HT); thus the actions of the neuropeptides were not mediated by cholinergic or serotonergic neurones. The stimulation of the musculature in the male reproductive tract and the inhibition of motility of the digestive system by FMRFamide and pQDPFLRFamide implicate these peptides in the control of reproductive behaviour. The effectiveness of pQDPFLRFamide in relaxing the retractor muscles and as a cardioexcitatory agent led to the hypothesis that this heptapeptide and FMRFamide, acting at distinct receptors, cooperate to regulate the excitability and contractility of the snail's musculature between the extremes of aestivation and active locomotion.


Development and external validation of a prediction model for venous thromboembolism in systemic lupus erythematosus.
Patients with systemic lupus erythematosus (SLE) have an increased risk of venous thromboembolism (VTE). We conducted this study to develop a risk score algorithm for VTE in patients with SLE that provides individualised risk estimates. We developed a clinical prediction model of VTE in 4502 patients with SLE based on the Chinese SLE Treatment and Research group cohort (CSTAR) from January 2009 to January 2020 and externally validated in 3780 patients with SLE in CSTAR from January 2020 to January 2022. Baseline data were obtained and VTE events were recorded during the follow-up. The prediction model was developed to predict VTE risk within 6 months in patients with SLE, using multivariate logistic regression and least absolute shrinkage and selection operator. SLE-VTE score and nomogram were established according to the model. A total of 4502 patients included in the development cohort, 135 had VTE events. The final prediction model (SLE-VTE score) included 11 variables: gender, age, body mass index, hyperlipidaemia, hypoalbuminaemia, C reactive protein, anti-β2GPI antibodies, lupus anticoagulant, renal involvement, nervous system involvement and hydroxychloroquine, with area under the curve of 0.947 and 0.808 in the development (n=4502) and external validation cohort (n=3780), respectively. According to the net benefit and predicted probability thresholds, we recommend annual screening of VTE in high risk (≥1.03%) patients with SLE. Various factors are related to the occurrence of VTE in patients with SLE. The proposed SLE-VTE risk score can accurately predict the risk of VTE and help identify patients with SLE with a high risk of VTE who may benefit from thromboprophylaxis.


Genomic characterisation of invasive non-typhoidal Salmonella enterica Subspecies enterica Serovar Bovismorbificans isolates from Malawi.
Invasive Non-typhoidal Salmonella (iNTS) are an important cause of bacteraemia in children and HIV-infected adults in sub-Saharan Africa. Previous research has shown that iNTS strains exhibit a pattern of gene loss that resembles that of host adapted serovars such as Salmonella Typhi and Paratyphi A. Salmonella enterica serovar Bovismorbificans was a common serovar in Malawi between 1997 and 2004. We sequenced the genomes of 14 Malawian bacteraemia and four veterinary isolates from the UK, to identify genomic variations and signs of host adaptation in the Malawian strains. Whole genome phylogeny of invasive and veterinary S. Bovismorbificans isolates showed that the isolates are highly related, belonging to the most common international S. Bovismorbificans Sequence Type, ST142, in contrast to the findings for S. Typhimurium, where a distinct Sequence Type, ST313, is associated with invasive disease in sub-Saharan Africa. Although genome degradation through pseudogene formation was observed in ST142 isolates, there were no clear overlaps with the patterns of gene loss seen in iNTS ST313 isolates previously described from Malawi, and no clear distinction between S. Bovismorbificans isolates from Malawi and the UK. The only defining differences between S. Bovismorbificans bacteraemia and veterinary isolates were prophage-related regions and the carriage of a S. Bovismorbificans virulence plasmid (pVIRBov). iNTS S. Bovismorbificans isolates, unlike iNTS S. Typhiumrium isolates, are only distinguished from those circulating elsewhere by differences in the mobile genome. It is likely that these strains have entered a susceptible population and are able to take advantage of this niche. There are tentative signs of convergent evolution to a more human adapted iNTS variant. Considering its importance in causing disease in this region, S. Bovismorbificans may be at the beginning of this process, providing a reference against which to compare changes that may become fixed in future lineages in sub-Saharan Africa.


Evaluation of Vaccine Safety After the First Public Sector Introduction of Typhoid Conjugate Vaccine-Navi Mumbai, India, 2018.
In December 2017, the World Health Organization (WHO) prequalified the first typhoid conjugate vaccine (TCV; Typbar-TCV). While no safety concerns were identified in pre- and postlicensure studies, WHO's Global Advisory Committee on Vaccine Safety recommended robust safety evaluation with large-scale TCV introductions. During July-August 2018, the Navi Mumbai Municipal Corporation (NMMC) launched the world's first public sector TCV introduction. Per administrative reports, 113 420 children 9 months-14 years old received TCV. We evaluated adverse events following immunization (AEFIs) using passive and active surveillance via (1) reports from the passive NMMC AEFI surveillance system, (2) telephone interviews with 5% of caregivers of vaccine recipients 48 hours and 7 days postvaccination, and (3) chart abstraction for adverse events of special interest (AESIs) among patients admitted to 5 hospitals using the Brighton Collaboration criteria followed by ascertainment of vaccination status. We identified 222/113 420 (0.2%) vaccine recipients with AEFIs through the NMMC AEFI surveillance system: 211 (0.19%) experienced minor AEFIs, 2 (0.002%) severe, and 9 serious (0.008%). At 48 hours postvaccination, 1852/5605 (33%) caregivers reported ≥1 AEFI, including injection site pain (n = 1452, 26%), swelling (n = 419, 7.5%), and fever (n = 416, 7.4%). Of the 4728 interviews completed at 7 days postvaccination, the most reported AEFIs included fever (n = 200, 4%), pain (n = 52, 1%), and headache (n = 42, 1%). Among 525 hospitalized children diagnosed with an AESI, 60 were vaccinated; no AESIs were causally associated with TCV. No unexpected safety signals were identified with TCV introduction. This provides further reassurance for the large-scale use of Typbar-TCV among children 9 months-14 years old.


Characteristics of Epstein-Barr virus envelope protein gp42.
Epstein-Barr virus (EBV) glycoprotein 42 (gp42) is a membrane protein essential for fusion and entry of EBV into host B-lymphocytes. Gp42 is a member of the protein-fold family C-type lectin or lectin-like domains (CLECT or CTLD) and specifically is classified as a natural-killer receptor (NKR)-like CLECT. Literature review and phylogenetic comparison show that EBV gp42 shares a common structure with other NKR-like CLECTs and possibly with many viral CTLDs, but does not appear to exhibit some common binding characteristics of many CTLDs, such as features required for calcium binding. The flexible N-terminal region adjacent to the CTLD fold is important for binding to other EBV glycoproteins and for a cleavage site that is necessary for infection of host cells. From structural studies of gp42 unbound and bound to receptor and extensive mutational analysis, a general model of how gp42 triggers membrane fusion utilizing both the flexible N-terminal region and the CTLD domain has emerged.


Sex-biased stress signaling: the corticotropin-releasing factor receptor as a model.
Sex differences in the prevalence or severity of many diseases and in the response to pharmacological agents are well recognized. Elucidating the biologic bases of these differences can advance our understanding of the pathophysiology of disease and facilitate the development of treatments. Despite the importance to medicine, this has been an area of limited research. Here, we review physiologic, cellular, and molecular findings supporting the idea that there are sex differences in receptor signaling and trafficking that can be determinants of pathology. The focus is on the receptor for corticotropin-releasing factor (CRF), the orchestrator of the stress response, which has been implicated in diverse stress-related diseases that show a female prevalence. Data are reviewed that show sex differences in the association of the CRF receptor (CRF1) with the Gs protein and β-arrestin 2 that would render females more responsive to acute stress and less able to adapt to chronic stress as a result of compromised CRF1 internalization. Because β-arrestin 2 serves to link CRF1 to Gs-independent signaling pathways, this sex-biased signaling is proposed to result in distinct cellular responses to stress that are translated to different physiologic and behavioral coping mechanisms and that can have different pathologic consequences. Because stress has been implicated in diverse medical and psychiatric diseases, these sex differences in CRF1 signaling could explain sex differences in a multitude of disorders. The possibility that analogous sex differences may occur with other G-protein-coupled receptors underscores the impact of this effect and is discussed.


Human ABC transporters at blood-CNS interfaces as determinants of CNS drug penetration.
Since the discovery of P-glycoprotein (P-gp) in brain microvessels composing the human blood-brain barrier (BBB), ATP-binding cassette (ABC) transporters have been recognized as bottlenecks in the development and delivery of neuropharmaceuticals. ABC transporters are expressed predominately at the plasma luminal membrane of brain capillary endothelial cells. These ABC transporters are responsible for the efflux of their substrates from the endothelial cells to the bloodstream against the concentration gradient and thus limit the entry of some drugs within the central nervous system (CNS). Advanced quantitative molecular biology tools allowed gene and protein quantification of the components of microvessels isolated from different species including human. Recently, positron emission tomography using radiolabelled probes that are substrates of ABC transporters allowed the determination of their functional activity at the human BBB. Here, we summarized new information regarding the relative expression, substrate recognition pattern for CNS drugs and functional activity of ABC transporters that are quantitatively expressed at the human BBB.


A Growth Mindset Frame Increases Opting In to Reading Information About Bias.
We explore the conditions under which people will opt in to reading information about bias and stereotypes, a key precursor to the types of self-directed learning that diversity and anti-bias advocates increasingly endorse. Across one meta-analysis (total N = 1,122; 7 studies, 5 pre-registered) and 2 pre-registered experiments (total N = 1,717), we identify a condition under which people opt in to reading more about implicit bias and stereotypes. People randomly assigned to read a growth, rather than fixed, mindset frame about bias opted in to read more information about stereotypes and implicit bias (Study 1 and Study 3). The mechanism that drove these effects was individuals' construal of the task as a challenge (Studies 2 and 3). Our findings offer insight into how to promote engagement with information about stereotypes and biases. We discuss how this work advances the study of mindsets and diversity science.


Microparticle-Induced Activation of the Vascular Endothelium Requires Caveolin-1/Caveolae.
Microparticles (MPs) are small membrane fragments shed from normal as well as activated, apoptotic or injured cells. Emerging evidence implicates MPs as a causal and/or contributing factor in altering normal vascular cell phenotype through initiation of proinflammatory signal transduction events and paracrine delivery of proteins, mRNA and miRNA. However, little is known regarding the mechanism by which MPs influence these events. Caveolae are important membrane microdomains that function as centers of signal transduction and endocytosis. Here, we tested the concept that the MP-induced pro-inflammatory phenotype shift in endothelial cells (ECs) depends on caveolae. Consistent with previous reports, MP challenge activated ECs as evidenced by upregulation of intracellular adhesion molecule-1 (ICAM-1) expression. ICAM-1 upregulation was mediated by activation of NF-κB, Poly [ADP-ribose] polymerase 1 (PARP-1) and the epidermal growth factor receptor (EGFR). This response was absent in ECs lacking caveolin-1/caveolae. To test whether caveolae-mediated endocytosis, a dynamin-2 dependent process, is a feature of the proinflammatory response, EC's were pretreated with the dynamin-2 inhibitor dynasore. Similar to observations in cells lacking caveolin-1, inhibition of endocytosis significantly attenuated MPs effects including, EGFR phosphorylation, activation of NF-κB and upregulation of ICAM-1 expression. Thus, our results indicate that caveolae play a role in mediating the pro-inflammatory signaling pathways which lead to EC activation in response to MPs.


Electroconvulsive therapy and determination of cerebral dominance.
Electroconvulsive therapy (ECT) often results in a number of short- and long-time side effects including memory impairment for past and current events, which can last for several months after ECT treatment. It has been suggested that unilateral ECT (uECT) with electrodes placed over the non-dominant (typically right) hemisphere significantly reduces side effects, especially memory disturbances. It is important to note that cerebral dominance equates to speech dominance and avoiding this area of the brain also reduces speech dysfunction after ECT. Traditionally, the routine clinical determination of cerebral dominance has been through the assessment of hand, foot and eye dominance, which is an easy and inexpensive approach that, however, does not ensure accuracy. This review of literature on different methods and techniques for determination of cerebral dominance and provides evidence that functional transcranial Doppler sonography (fTCD) represents a valid and safe alternative to invasive techniques for identifying speech lateralisation. It can be concluded that fTCD, notwithstanding its costs, could be used as a standard procedure prior to uECT treatment to determine cerebral dominance, thereby further reducing cognitive side-effects of ECT and possibly making it more acceptable to both patients and clinicians.


[Study of propoleum dosage in Peyronie's disease].
To determine the optimal dose for propoleum caps in Peyronie's disease. We evaluated two groups of 17 patients each, analyzing pain and plaque size on physical and ultrasound examination, before and after treatment. Group A patients received caps with 300 mg of propoleum powder daily for six months, and Group B 900 mg. Predominant ages were between 41 and 70 years. More than half the patients in group A had pain; more than half the patients in group B did not have pain; considering both groups there were more patients without pain. Physical reduction of the plaque: Group A: 0.6 cm, Group B: 0.8 cm. Ultrasound reduction of the plaque: Group A: 1.3 cm, Group B: 0.8 cm. In the higher dose group (B) clinical and ultrasound improvement started earlier. 79.4% of the treated patients evaluated preferred improvement or slight improvement, being this result statistically significant. Pain is not always an accompanying symptom in Peyronie's disease. Both doses administered reduced the size of the plaque, but the higher one has an earlier improvement. We may consider propoleum administered in this way another treatment option for this disease, with good results and low cost.


A formal analysis of stopping criteria of decomposition methods for support vector machines.
In a previous paper, the author (2001) proved the convergence of a commonly used decomposition method for support vector machines (SVMs). However, there is no theoretical justification about its stopping criterion, which is based on the gap of the violation of the optimality condition. It is essential to have the gap asymptotically approach zero, so we are sure that existing implementations stop in a finite number of iterations after reaching a specified tolerance. Here, we prove this result and illustrate it by two extensions: /spl nu/-SVM and a multiclass SVM by Crammer and Singer (2001). A further result shows that, in final iterations of the decomposition method, only a particular set of variables are still being modified. This supports the use of the shrinking and caching techniques in some existing implementations. Finally, we prove the asymptotic convergence of a decomposition method for this multiclass SVM. Discussions on the difference between this convergence proof and the one in another paper by Lin are also included.


Preparation of Formalin-fixed Paraffin-embedded Tissue Cores for both RNA and DNA Extraction.
Formalin-fixed paraffin embedded tissue (FFPET) represents a valuable, well-annotated substrate for molecular investigations. The utility of FFPET in molecular analysis is complicated both by heterogeneous tissue composition and low yields when extracting nucleic acids. A literature search revealed a paucity of protocols addressing these issues, and none that showed a validated method for simultaneous extraction of RNA and DNA from regions of interest in FFPET. This method addresses both issues. Tissue specificity was achieved by mapping cancer areas of interest on microscope slides and transferring annotations onto FFPET blocks. Tissue cores were harvested from areas of interest using 0.6 mm microarray punches. Nucleic acid extraction was performed using a commercial FFPET extraction system, with modifications to homogenization, deparaffinization, and Proteinase K digestion steps to improve tissue digestion and increase nucleic acid yields. The modified protocol yields sufficient quantity and quality of nucleic acids for use in a number of downstream analyses, including a multi-analyte gene expression platform, as well as reverse transcriptase coupled real time PCR analysis of mRNA expression, and methylation-specific PCR (MSP) analysis of DNA methylation.


[Angiodysplasia of the large intestine: unusual rectal localization].
The authors report a case of lower gastrointestinal hemorrhage in a 69-year-old male, in which the sequence colonoscopy-angiography identified the source of bleeding as a rare Angiodysplasia (AD) of the rectum. Such vascular abnormalities are one of the most common causes of major lower gastrointestinal tract bleeding in the elderly and usually occurs predominantly in the right side of the colon. The clinical presentation of Angiodysplasia is varied and accurate diagnosis usually requires a combination of diagnostic techniques such as colonoscopy and angiography. The optimal management is uncertain and should be individualized for each patient depending on severity and rate of rebleeding. A conservative medical approach is indicated for many patients, while endoscopic treatment does not seem modify the risk of recurrent bleeding. In case of massive hemorrhage or recurrent bleeding surgery still represents the definitive treatment for Angiodysplasia. However the risk of rebleeding following surgery is a considerable problem and varies in literature from 5% to 30%.


A high-resolution structure of the pre-microRNA nuclear export machinery.
Nuclear export of microRNAs (miRNAs) by exportin-5 (Exp-5) is an essential step in miRNA biogenesis. Here, we present the 2.9 angstrom structure of the pre-miRNA nuclear export machinery formed by pre-miRNA complexed with Exp-5 and a guanine triphosphate (GTP)-bound form of the small nuclear guanine triphosphatase (GTPase) Ran (RanGTP). The x-ray structure shows that Exp-5:RanGTP recognizes the 2-nucleotide 3' overhang structure and the double-stranded stem of the pre-miRNA. Exp-5:RanGTP shields the pre-miRNA stem from degradation in a baseball mitt-like structure where it is held by broadly distributed weak interactions, whereas a tunnel-like structure of Exp-5 interacts strongly with the 2-nucleotide 3' overhang through hydrogen bonds and ionic interactions. RNA recognition by Exp-5:RanGTP does not depend on RNA sequence, implying that Exp-5:RanGTP can recognize a variety of pre-miRNAs.


Towards a correlative approach for characterising single virus particles by transmission electron microscopy and nanoscale Raman spectroscopy.
The morphology and structure of biological nanoparticles, such as viruses, can be efficiently analysed by transmission electron microscopy (TEM). To chemically characterise such nanoparticles in heterogeneous samples at the single particle level, we suggest tip-enhanced Raman spectroscopy (TERS) as a correlative method. Here we describe a TERS-compatible staining procedure for TEM which involves sample pre-scanning by TEM imaging, nanoparticle relocalisation by atomic force microscopy (AFM) followed by spectroscopic characterization of the virus nanoparticles using TERS. First successful correlative measurements are demonstrated on tobacco mosaic virus particles deposited on silicon-based TEM sample supports. In addition, the advantages and problems of this methodology are discussed.


Antibiotic-Impregnated Central Venous Catheters Do Not Change Antibiotic Resistance Patterns.
Antibiotic-impregnated central venous catheters (CVCs) decrease the incidence of infection in high-risk patients. However, use of these catheters carries the hypothetical risk of inducing antibiotic resistance. We hypothesized that routine use of minocycline and rifampin-impregnated catheters (MR-CVC) in a single intensive care unit (ICU) would change the resistance profile for Staphylococcus aureus. We reviewed antibiotic susceptibilities of S. aureus isolates obtained from blood cultures in a large urban teaching hospital from 2002-2015. Resistance patterns were compared before and after implementation of MR-CVC use in the surgical ICU (SICU) in August 2006. We also compared resistance patterns of S. aureus obtained in other ICUs and in non-ICU patients, in whom MR-CVCs were not used. Data for rifampin, oxacillin, and clindamycin were available for 9,703 cultures; tetracycline resistance data were available for 4,627 cultures. After implementation of MR-CVC use in the SICU, rifampin resistance remained unchanged, with rates the same as in other ICU and non-ICU populations (3%). After six years of use of MR-CVCs in the SICU, the rate of tetracycline resistance was unchanged in all facilities (1%-3%). The use of MR-CVCs was not associated with any change in S. aureus oxacillin-resistance rates in the SICU (66% vs. 60%). However, there was a significant decrease in S. aureus clindamycin resistance (59% vs. 34%; p < 0.05) in SICU patients. Routine use of rifampin-minocycline-impregnated CVCs in the SICU was not associated with increased resistance of S. aureus isolates to rifampin or tetracyclines.


Phenotype Prediction from Metagenomic Data Using Clustering and Assembly with Multiple Instance Learning (CAMIL).
The recent advent of Metagenome Wide Association Studies (MGWAS) provides insight into the role of microbes on human health and disease. However, the studies present several computational challenges. In this paper, we demonstrate a novel, efficient, and effective Multiple Instance Learning (MIL) based computational pipeline to predict patient phenotype from metagenomic data. MIL methods have the advantage that besides predicting the clinical phenotype, we can infer the instance level label or role of microbial sequence reads in the specific disease. Specifically, we use a Bag of Words method, which has been shown to be one of the most effective and efficient MIL methods. This involves assembly of the metagenomic sequence data, clustering of the assembled contigs, extracting features from the contigs, and using an SVM classifier to predict patient labels and identify the most relevant sequence clusters. With the exception of the given labels for the patients, this entire process is de novo (unsupervised). We call our pipeline "CAMIL", which stands for Clustering and Assembly with Multiple Instance Learning. We use multiple state-of-the-art clustering methods for feature extraction, evaluation, and comparison of the performance of our proposed approach for each of these clustering methods. We also present a fast and scalable pre-clustering algorithm as a preprocessing step for our proposed pipeline. Our approach achieves efficiency by partitioning the large number of sequence reads into groups (called canopies) using locality sensitive hashing (LSH). These canopies are then refined by using state-of-the-art sequence clustering algorithms. We use data from a well-known MGWAS study of patients with Type-2 Diabetes and show that our pipeline significantly outperforms the classifier used in that paper, as well as other common MIL methods.


Microfabricated polymer devices for automated sample delivery of peptides for analysis by electrospray ionization tandem mass spectrometry.
Delivery of proteins and peptides to electrospray ionization mass spectrometers (ESI-MS) has been demonstrated using glass and quartz microfabricated devices. This paper reports the construction and use of poly(dimethylsiloxane) (PDMS) microfabricated soft polymer devices with mass spectrometry for protein analysis. The PDMS devices were fabricated using replica molding against a patterned photoresist generated by photolithographic techniques. The PDMS devices were connected to the mass spectrometer via a derivatized transfer capillary and samples were transferred by electroosmotic pumping. The formulation of PDMS was optimized for compatibility with ESI, and the devices were tested for performance. The practical application of PDMS devices was demonstrated by the identification of rat serum albumin separated by 2-D gel electrophoresis. Extended contact of the sample with the surface of the PDMS device did not significantly affect the sample analysis, and the limit of detection for samples run on a PDMS device was comparable to the limit of detection achieved on glass devices. This study suggests that PDMS devices fabricated using replica molding are compatible with ESI-MS. This will potentially lead to the construction of inexpensive microfabricated devices with complex designs and advanced functionalities.


Antivascular endothelial growth factor receptor (fetal liver kinase 1) monoclonal antibody inhibits tumor angiogenesis and growth of several mouse and human tumors.
Tumor angiogenesis is mediated by tumor-secreted angiogenic growth factors that interact with their surface receptors expressed on endothelial cells. Vascular endothelial growth factor (VEGF) and its receptor [fetal liver kinase 1 (Flk-1)/kinase insert domain-containing receptor] play an important role in vascular permeability and tumor angiogenesis. Previously, we reported on the development of anti-Flk-1 and antikinase insert domain-containing receptor monoclonal antibodies (mAbs) that potently inhibit VEGF binding and receptor signaling. Here, we report the effect of anti-Flk-1 mAb (DC101) on angiogenesis and tumor growth. Angiogenesis in vivo was examined using a growth factor supplemented (basic fibroblast growth factor + VEGF) Matrigel plug and an alginate-encapsulated tumor cell (Lewis lung) assay in C57BL/6 mice. Systemic administration of DC101 every 3 days markedly reduced neovascularization of Matrigel plugs and tumor-containing alginate beads in a dose-dependent fashion. Histological analysis of Matrigel plugs showed reduced numbers of endothelial cells and vessel structures. Several mouse tumors and human tumor xenografts in athymic mice were used to examine the effect of anti-Flk-1 mAb treatment on tumor angiogenesis and growth. Anti-Flk-1 mAb treatment significantly suppressed the growth of primary murine Lewis lung, 4T1 mammary, and B16 melanoma tumors and growth of Lewis lung metastases. DC101 also completely inhibited the growth of established epidermoid, glioblastoma, pancreatic, and renal human tumor xenografts. Histological examination of anti-Flk-1 mAb-treated tumors showed evidence of decreased microvessel density, tumor cell apoptosis, decreased tumor cell proliferation, and extensive tumor necrosis. These findings support the conclusion that anti-Flk-1 mAb treatment inhibits tumor growth by suppression of tumor-induced neovascularization and demonstrate the potential for therapeutic application of anti-VEGF receptor antibody in the treatment of angiogenesis-dependent tumors.


In vivo role of catalase-peroxidase in synechocystis sp. strain PCC 6803.
The katG gene coding for the only catalase-peroxidase in the cyanobacterium Synechocystis sp. strain PCC 6803 was deleted in this organism. Although the rate of H2O2 decomposition was about 30 times lower in the DeltakatG mutant than in the wild type, the strain had a normal phenotype and its doubling time as well as its resistance to H2O2 and methyl viologen were indistinguishable from those of the wild type. The residual H2O2-scavenging capacity was more than sufficient to deal with the rate of H2O2 production by the cell, estimated to be less than 1% of the maximum rate of photosynthetic electron transport in vivo. We propose that catalase-peroxidase has a protective role against environmental H2O2 generated by algae or bacteria in the ecosystem (for example, in mats). This protective role is most apparent at a high cell density of the cyanobacterium. The residual H2O2-scavenging activity in the DeltakatG mutant was a light-dependent peroxidase activity. However, neither glutathione peroxidase nor ascorbate peroxidase accounted for a significant part of this H2O2-scavenging activity. When a small thiol such as dithiothreitol was added to the medium, the rate of H2O2 decomposition in the DeltakatG mutant increased more than 10-fold, indicating that a thiol-specific peroxidase, for which thioredoxin may be the physiological electron donor, is present. Oxidized thioredoxin is likely to be reduced again by photosynthetic electron transport. Therefore, under laboratory conditions, there are only two enzymatic mechanisms for H2O2 decomposition present in Synechocystis sp. strain PCC 6803. One is catalyzed by a catalase-peroxidase, and the other is catalyzed by thiol-specific peroxidase.


An analysis of the palliative care education needs of RGNs and HCAs in nursing homes in Ireland.
To identify the palliative care education needs of registered general nurses (RGNs) and health-care assistants (HCAs) working in care of the older person units (nursing homes). A questionnaire was sent to all RGNs and HCAs working in nursing homes run by the Health Service Executive (HSE) in one health-care region in Ireland. RGNs identified eight specific educational needs: understanding of the dying process; pain and symptom management; improved communication skills; nutritional assessment issues; practical skills; the appropriate use and management of syringe drivers; and the role of complementary therapies. HCAs identified a need to improve their communication skills and clarify their role. Both groups identified the need for support when caring for dying patients and the need to improve their understanding of palliative care and the role of specialist palliative care (SPC) services. There was a significant difference between the RGNs' and the HCAs' level of understanding of palliative care (P = 0.00). This study demonstrated that RGNs and HCAs working in care of the older person settings have palliative care educational needs. It also established a difference in the educational background and palliative care understanding of RGNs and HCAs. This study therefore identified the need to develop separate education programmes.


Direct near-field optical imaging of higher order plasmonic resonances.
We map in real space and by purely optical means near-field optical information of localized surface plasmon polariton (LSPP) resonances excited in nanoscopic particles. We demonstrate that careful polarization control enables apertureless scanning near-field optical microscopy (aSNOM) to image dipolar and quadrupolar LSPPs of the bare sample with high fidelity in both amplitude and phase. This establishes a routine method for in situ optical microscopy of plasmonic and other resonant structures under ambient conditions.


Stimulation by forskolin of the thyroid adenylate cyclase, cyclic AMP accumulation and iodine metabolism.
Forskolin, a diterpene hypotensive drug, activates adenylate cyclase in brain and in some other tissues (Seamon et al., 1981). Forskolin activated adenylate cyclase in particulate preparations and enhanced cyclic AMP accumulation in slices of dog thyroid. These effects were maximal within minutes and remained constant afterwards. The action of forskolin on intact cells disappeared rapidly after washing. It reproduced two known cyclic AMP-mediated TSH effects: the activation of secretion and of protein iodination. Forskolin thus provides a very convenient tool for the study of the action of defined elevations of cyclic AMP level in thyroid cells. The activation by forskolin of adenylate cyclase was not reduced by Mn2+ which uncouples TSH and PGE1 action. This suggests that in the thyroid also, forskolin acts beyond the receptor level. The effect of forskolin on cyclic AMP accumulation was inhibited by the known negative regulators of this system in the thyroid, acetylcholine, iodide, norepinephrine, PGF1 alpha and adenosine. On the other hand, forskolin potentiated the effects of TSH, PGE1 and cholera toxin. These data show that, though it does not require the receptors for its action, forskolin does not uncouple them from the catalytic unit of adenylate cyclase.


Androgen receptor signaling promotes Treg suppressive function during allergic airway inflammation.
Women have higher prevalence of asthma compared with men. In asthma, allergic airway inflammation is initiated by IL-33 signaling through ST2, leading to increased IL-4, IL-5, and IL-13 production and eosinophil infiltration. Foxp3+ Tregs suppress and ST2+ Tregs promote allergic airway inflammation. Clinical studies showed that the androgen dehydroepiandrosterone (DHEA) reduced asthma symptoms in patients, and mouse studies showed that androgen receptor (AR) signaling decreased allergic airway inflammation. Yet the impact of AR signaling on lung Tregs remains unclear. Using AR-deficient and Foxp3 fate-mapping mice, we determined that AR signaling increased Treg suppression during Alternaria extract (Alt Ext; allergen) challenge by stabilizing Foxp3+ Tregs and limiting the number of ST2+ ex-Tregs and IL-13+ Th2 cells and ex-Tregs. AR signaling also decreased Alt Ext-induced ST2+ Tregs in mice by limiting expression of Gata2, a transcription factor for ST2, and by decreasing Alt Ext-induced IL-33 production from murine airway epithelial cells. We confirmed our findings in human cells where 5α-dihydrotestosterone (DHT), an androgen, decreased IL-33-induced ST2 expression in lung Tregs and decreased Alt Ext-induced IL-33 secretion in human bronchial epithelial cells. Our findings showed that AR signaling stabilized Treg suppressive function, providing a mechanism for the sex difference in asthma.


Personizing the prediction of future susceptibility to a specific disease.
A traceable biomarker is a member of a disease's molecular pathway. A disease may be associated with several molecular pathways. Each different combination of these molecular pathways, to which detected traceable biomarkers belong, may serve as an indicative of the elicitation of the disease at a different time frame in the future. Based on this notion, we introduce a novel methodology for personalizing an individual's degree of future susceptibility to a specific disease. We implemented the methodology in a working system called Susceptibility Degree to a Disease Predictor (SDDP). For a specific disease d, let S be the set of molecular pathways, to which traceable biomarkers detected from most patients of d belong. For the same disease d, let S' be the set of molecular pathways, to which traceable biomarkers detected from a certain individual belong. SDDP is able to infer the subset S'' ⊆{S-S'} of undetected molecular pathways for the individual. Thus, SDDP can infer undetected molecular pathways of a disease for an individual based on few molecular pathways detected from the individual. SDDP can also help in inferring the combination of molecular pathways in the set {S'+S''}, whose traceable biomarkers collectively is an indicative of the disease. SDDP is composed of the following four components: information extractor, interrelationship between molecular pathways modeler, logic inferencer, and risk indicator. The information extractor takes advantage of the exponential increase of biomedical literature to automatically extract the common traceable biomarkers for a specific disease. The interrelationship between molecular pathways modeler models the hierarchical interrelationships between the molecular pathways of the traceable biomarkers. The logic inferencer transforms the hierarchical interrelationships between the molecular pathways into rule-based specifications. It employs the specification rules and the inference rules for predicate logic to infer as many as possible undetected molecular pathways of a disease for an individual. The risk indicator outputs a risk indicator value that reflects the individual's degree of future susceptibility to the disease. We evaluated SDDP by comparing it experimentally with other methods. Results revealed marked improvement.


Determinants associated with the utilization of primary and specialized mental health services.
The study aims to compare variables associated with the exclusive and joint use of primary and specialized care for mental health reasons by individuals diagnosed with a mental disorder in a Montreal/Canadian catchment area. Data were collected from a random sample (2,443 individuals). Among 406 people, diagnosed with a mental disorder 12 months pre-interview, 212 (52%) reported having used healthcare services. Compared to users of primary care only, people who sought both primary and specialized care presented more mental disorders and lower quality of life. People using only specialized healthcare received significantly less social support than persons using primary care exclusively and lived in neighborhoods with a high proportion of rental housing. Healthcare service provision should favor social networking and enable social cohesion and integration, particularly in neighborhoods with a high proportion of rental housing. Shared care and enhanced collaboration with other public and community-based resources should be encouraged.


Botulinum toxin assessment, intervention and after-care for upper limb hypertonicity in adults: international consensus statement.
Upper limb spasticity affecting elbow, wrist, and finger flexors can be safely and effectively reduced with injections of botulinum toxin type-A (BoNT-A). It has been best studied in adults in the context of post-stroke spasticity. The clinical benefits include reduction in pain and deformity, improvement in washing and dressing the upper limb, and a reduction in caregiver burden (Class I evidence, recommendation level A). Some patients show improvement in function performed by active movement of the affected upper limb (Class III evidence, recommendation C), but predicting and measuring this is difficult, and further research is needed. An individually based approach to treatment and outcome measurement is preferred (Class IV, recommendation U). More research is needed to resolve many unknown issues of assessment and treatment, using research methods appropriate to the question.


When co-action eliminates the Simon effect: disentangling the impact of co-actor's presence and task sharing on joint-task performance.
This study aimed at assessing whether the mere belief of performing a task with another person, who is in charge of the complementary part of the task, is sufficient for the so-called joint Simon effect to occur. In all three experiments of the study, participants sat alone in a room and underwent two consecutive Go/NoGo tasks that were identical except for the instructions. In Experiment 1, participants performed the task first individually (baseline task), and then either co-acting with another person who responded from an unknown location to the NoGo stimuli (joint task) or imaging themselves responding to the NoGo stimuli (imaginative task). Relative to the baseline, the instructions of the imaginative task made the Simon effect occur, while those of the joint task were ineffective in eliciting the effect. This result suggests that sharing a task with a person who is known to be in charge of the complementary task, but is not physically present, is not sufficient to induce the representation of an alternative response able to produce interference, which happens instead when the instructions explicitly require to imagine such a response. Interestingly, we observed that when the Simon effect was already present in the baseline task (i.e., when the response alternative to the Go response was represented in the individual task due to non-social factors), it disappeared in the joint task. We propose that, when no information about the co-actor's position is available, the division of labor between the participant and co-actor allows participants to filter out the possible (incidental) representation of the alternative response from their task representation, thus eliminating potential sources of interference. This account is supported by the results of Experiments 2 and 3 and suggests that under certain circumstances task-sharing may reduce the interference produced by the irrelevant information, rather than increase it.


Acute ischaemic stroke associated with SARS-CoV-2 infection in North America.
To analyse the clinical characteristics of COVID-19 with acute ischaemic stroke (AIS) and identify factors predicting functional outcome. Multicentre retrospective cohort study of COVID-19 patients with AIS who presented to 30 stroke centres in the USA and Canada between 14 March and 30 August 2020. The primary endpoint was poor functional outcome, defined as a modified Rankin Scale (mRS) of 5 or 6 at discharge. Secondary endpoints include favourable outcome (mRS ≤2) and mortality at discharge, ordinal mRS (shift analysis), symptomatic intracranial haemorrhage (sICH) and occurrence of in-hospital complications. A total of 216 COVID-19 patients with AIS were included. 68.1% (147/216) were older than 60 years, while 31.9% (69/216) were younger. Median [IQR] National Institutes of Health Stroke Scale (NIHSS) at presentation was 12.5 (15.8), and 44.2% (87/197) presented with large vessel occlusion (LVO). Approximately 51.3% (98/191) of the patients had poor outcomes with an observed mortality rate of 39.1% (81/207). Age >60 years (aOR: 5.11, 95% CI 2.08 to 12.56, p<0.001), diabetes mellitus (aOR: 2.66, 95% CI 1.16 to 6.09, p=0.021), higher NIHSS at admission (aOR: 1.08, 95% CI 1.02 to 1.14, p=0.006), LVO (aOR: 2.45, 95% CI 1.04 to 5.78, p=0.042), and higher NLR level (aOR: 1.06, 95% CI 1.01 to 1.11, p=0.028) were significantly associated with poor functional outcome. There is relationship between COVID-19-associated AIS and severe disability or death. We identified several factors which predict worse outcomes, and these outcomes were more frequent compared to global averages. We found that elevated neutrophil-to-lymphocyte ratio, rather than D-Dimer, predicted both morbidity and mortality.


Biomimetic platforms for human stem cell research.
Stem cells are central to developing new treatment options for tissue regeneration and constructing controllable models for biological research. Bioengineered cell culture environments that combine microenvironmental control with tissue-specific transport and signaling are critical tools in our efforts to study tissue development, regeneration, and disease under conditions that predict the human in vivo context. We propose that experimentation at the interfaces of biology, engineering, and medical sciences is critical for unlocking the full potential of stem cells. Here, we focus on the design and utilization of in vitro platforms that recapitulate the environments associated with tissue development, disease, and regeneration.


Epidemiology of ovarian cancer: a review.
Ovarian cancer (OC) is the seventh most commonly diagnosed cancer among women in the world and the tenth most common in China. Epithelial OC is the most predominant pathologic subtype, with five major histotypes that differ in origination, pathogenesis, molecular alterations, risk factors, and prognosis. Genetic susceptibility is manifested by rare inherited mutations with high to moderate penetrance. Genome-wide association studies have additionally identified 29 common susceptibility alleles for OC, including 14 subtype-specific alleles. Several reproductive and hormonal factors may lower risk, including parity, oral contraceptive use, and lactation, while others such as older age at menopause and hormone replacement therapy confer increased risks. These associations differ by histotype, especially for mucinous OC, likely reflecting differences in etiology. Endometrioid and clear cell OC share a similar, unique pattern of associations with increased risks among women with endometriosis and decreased risks associated with tubal ligation. OC risks associated with other gynecological conditions and procedures, such as hysterectomy, pelvic inflammatory disease, and polycystic ovarian syndrome, are less clear. Other possible risk factors include environmental and lifestyle factors such as asbestos and talc powder exposures, and cigarette smoking. The epidemiology provides clues on etiology, primary prevention, early detection, and possibly even therapeutic strategies.


The assembly of HIV within the Golgi apparatus and Golgi-derived vesicles of JM cell syncytia.
Chronic infection of the T-lymphocyte cell line JM with HIV-1 isolate GB8 results in the formation of multinucleate cells (syncytia). Transmission electron microscopy of these syncytia showed the presence of HIV particles both at the cell surface and within cytoplasmic vesicles. HIV particles were observed in dilated Golgi cisternae and Golgi-derived vesicles and in large vacuoles near the periphery of the syncytia. Immunolabelling was performed using an affinity-purified antiserum to the Golgi enzyme galactosyltransferase. This enzyme was consistently localized within both the Golgi apparatus and within virus-containing vesicles of JM syncytia, indicating that these vesicles originated from the Golgi apparatus.


Assay of prolyl 4-hydroxylase by the chromatographic determination of [14C]succinic acid on ion-exchange minicolumns.
An assay for prolyl 4-hydroxylase (EC 18.104.22.168) is described which measures succinic acid produced during the decarboxylation of 2-oxoglutaric acid in the presence of poly(L-Pro-Gly-L-Pro). [1-14C]Succinic acid was separated from its precursor 2-oxo[5-14C]glutaric acid by using ion-exchange minicolumns. The contamination of succinic acid by 2-oxoglutaric acid was approx. 1%, and the recovery of succinic acid was 100%. Kinetic parameters of prolyl 4-hydroxylase measured by the assay showed good agreement with published values. Our experience indicates that the measurement of prolyl 4-hydroxylase by the production of succinic acid is especially suited to investigations involving large numbers of assays.


Mechanism in the reaction of cytochrome c oxidase with organic hydroperoxides: an ESR spin-trapping investigation.
Organic hydroperoxides are of great utility in probing the reaction mechanism and the toxicological consequences of lipid peroxidation. In the present study, ESR spin-trapping was employed to investigate the peroxidation of mitochondrial cytochrome c oxidase (CcO) with t-butyl hydroperoxide (t-BuOOH) and cumene hydroperoxide (CumOOH). The spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) was used to detect the radical species formed from the reaction of CcO with t-BuOOH. The presence of t-BuOOH-derived alkoxyl radical (t-BuO*) as the primary radical indicates reductive scission of the O-O bond by CcO. The ESR signal of DMPO/*Ot-Bu can be partially abolished by cyanide, implying that the reductive cleavage involved the haem a(3)Cu(B) binuclear site of CcO. A nitroso spin trap, 2-methyl-2-nitrosopropane (MNP), was used to detect and identify radical species from the reaction of CcO with CumOOH. In addition to the t-BuOOH-derived methyl, hydroxylmethyl and tertiary carbon-centred radicals, a protein-derived radical was detected. The intensity of the ESR signal from the protein radical increased with the CumOOH concentration at low CumOOH/CcO ratios, with maximal intensity at a ratio of 100 mol of CumOOH/mol of CcO. The immobilized protein radical adduct of MNP was stable and persistent after dialysis; it was also resistant to proteolytic digestion, suggesting that it was formed in the transmembrane region, a region that is not accessible to proteases. Its signal was greatly enhanced when CcO cysteine residues were chemically modified by N-ethylmaleimide, when the tryptophan residues in CcO were oxidized by N-bromosuccimide, and when tyrosine residues on the surface of CcO were iodinated, showing that a radical equilibrium was established among the cysteine, tryptophan and tyrosine residues of the protein-centred radical. Pre-treatment of CcO with cyanide prevented detectable MNP adduct formation, confirming that the haem a(3)-Cu(B) binuclear centre was the initial reaction site. When the CcO was pre-treated with 10 mM (100 equivalents) of CumOOH, the enzyme activity decreased by more than 20%. This inhibition was persistent after dialysis, suggesting that the detected protein-centred radical was, in part, involved in the irreversible inactivation by CumOOH. Visible spectroscopic analysis revealed that the haem a of CcO was not affected during the reaction. However, the addition of pyridine to the reaction mixture under alkaline conditions resulted in the destruction of the haem centre of CcO, suggesting that its protein matrix rather than its haem a is the target of oxidative damage by the organic hydroperoxide.


Mitochondrial DNA deletion mutations: a causal role in sarcopenia.
Mitochondrial DNA (mtDNA) deletion mutations accumulate with age in tissues of a variety of species. Although the relatively low calculated abundance of these deletion mutations in whole tissue homogenates led some investigators to suggest that these mutations do not have any physiological impact, their focal and segmental accumulation suggests that they can, and do, accumulate to levels sufficient to affect the metabolism of a tissue. This phenomenon is most clearly demonstrated in skeletal muscle, where the accumulation of mtDNA deletion mutations remove critical subunits that encode for the electron transport system (ETS). In this review, we detail and provide evidence for a molecular basis of muscle fiber loss with age. Our data suggest that the mtDNA deletion mutations, which are generated in tissues with age, cause muscle fiber loss. Within a fiber, the process begins with a mtDNA replication error, an error that results in a loss of 25-80% of the mitochondrial genome. This smaller genome is replicated and, through a process not well understood, eventually comprises the majority of mtDNA within the small affected region of the muscle fiber. The preponderance of the smaller genomes results in a dysfunctional ETS in the affected area. As a consequence of both the decline in energy production and the increase in oxidative damage in the region, the fiber is no longer capable of self-maintenance, resulting in the observed intrafiber atrophy and fiber breakage. We are therefore proposing that a process contained within a very small region of a muscle fiber can result in breakage and loss of muscle fiber from the tissue.


Prevalence of COVID-19 and Psychotropic Drug Treatment in Psychiatric In-patients in Germany in 2020: Results from a Nationwide Pilot Survey.
In patients with a pre-existing mental disorder, an increased risk for a first manifestation of a psychiatric disorder in COVID-19 patients, a more severe course of COVID-19 and an increased mortality have been described. Conversely, observations of lower COVID-19 incidences in psychiatric in-patients suggested protective effects of psychiatric treatment and/or psychotropic drugs against COVID-19. A retrospective multi-center study was conducted in 24 German psychiatric university hospitals. Between April and December 2020 (the first and partly second wave of COVID-19), the effects of COVID-19 were assessed on psychiatric in-patient care, the incidence and course of a SARS-CoV-2 infection, and treatment with psychotropic drugs. Patients (n=36,322) were admitted to the hospitals. Mandatory SARS-CoV-2 tests before/during admission were reported by 23 hospitals (95.8%), while 18 (75%) conducted regular testing during the hospital stay. Two hundred thirty-two (0.6%) patients were tested SARS-CoV-2-positive. Thirty-seven (16%) patients were receiving medical treatment for COVID-19 at the psychiatric hospital, ten (4.3%) were transferred to an intermediate/intensive care unit, and three (1.3%) died. The most common prescription for SARS-CoV-2-positive patients was for second-generation antipsychotics (n=79, 28.2%) and antidepressants (SSRIs (n=38, 13.5%), mirtazapine (n=36, 12.9%) and SNRIs (n=29, 10.4%)). Contrary to previous studies, our results showed a low number of infections and mortality in SARS-CoV-2-positive psychiatric patients. Several preventive measures seem effective to protect this vulnerable group. Our observations are compatible with the hypothesis of a protective effect of psychotropic drugs against COVID-19 as the overall mortality and need for specific medical treatment was low.


Co-Creating and Evaluating an App-Based Well-Being Intervention: The HOW (Healthier Outcomes at Work) Social Work Project.
Stress and mental health at work are the leading causes of long-term sickness absence in the UK, with chronically poor working conditions impacting employee physiological and psychological health. Social workers play a significant part in the fabric of UK society, but have one of the most stressful occupations in the country. The aim of this project was to work with UK social workers to co-develop, implement, and evaluate a series of smartphone-based mental health initiatives. A Participatory Action Research (PAR) approach, consisting of semi-structured interviews and focus group and steering group discussions, was utilized to design the mental health and well-being interventions. Study efficacy was evaluated via a pre- and post-intervention survey and post-intervention semi-structured interviews. Interventions developed were psycho-educational, improved top-down and bottom-up communication, and provided access to a Vocational Rehabilitation Assistant for those struggling and at risk of sickness absence. Six months following dissemination, surveys demonstrated significant improvements in communication, and mean score improvements in four other working conditions. This project, therefore, demonstrates that co-developed initiatives can be positively impactful, despite post-intervention data collection being impacted by COVID-19. Future studies should build upon these findings and broaden the PAR approach nationally while taking a robust approach to evaluation.


Microfluidic-based production of [68Ga]Ga-FAPI-46 and [68Ga]Ga-DOTA-TOC using the cassette-based iMiDEV™ microfluidic radiosynthesizer.
The demand for 68Ga-labeled radiotracers has significantly increased in the past decade, driven by the development of diversified imaging tracers, such as FAPI derivatives, PSMA-11, DOTA-TOC, and DOTA-TATE. These tracers have exhibited promising results in theranostic applications, fueling interest in exploring them for clinical use. Among these probes, 68Ga-labeled FAPI-46 and DOTA-TOC have emerged as key players due to their ability to diagnose a broad spectrum of cancers ([68Ga]Ga-FAPI-46) in late-phase studies, whereas [68Ga]Ga-DOTA-TOC is clinically approved for neuroendocrine tumors. To facilitate their production, we leveraged a microfluidic cassette-based iMiDEV radiosynthesizer, enabling the synthesis of [68Ga]Ga-FAPI-46 and [68Ga]Ga-DOTA-TOC based on a dose-on-demand (DOD) approach. Different mixing techniques were explored to influence radiochemical yield. We achieved decay-corrected yield of 44 ± 5% for [68Ga]Ga-FAPI-46 and 46 ± 7% for [68Ga]Ga-DOTA-TOC in approximately 30 min. The radiochemical purities (HPLC) of [68Ga]Ga-FAPI-46 and [68Ga]Ga-DOTA-TOC were 98.2 ± 0.2% and 98.4 ± 0.9%, respectively. All the quality control results complied with European Pharmacopoeia quality standards. We optimized various parameters, including 68Ga trapping and elution, cassette batches, passive mixing in the reactor, and solid-phase extraction (SPE) purification and formulation. The developed synthesis method reduced the amount of precursor and other chemicals required for synthesis compared to conventional radiosynthesizers. The microfluidic-based approach enabled the implementation of radiosynthesis of [68Ga]Ga-FAPI-46 and [68Ga]Ga-DOTA-TOC on the iMiDEV™ microfluidic module, paving the way for their use in preclinical and clinical applications. The microfluidic synthesis approach utilized 2-3 times less precursor than cassette-based conventional synthesis. The synthesis method was also successfully validated in a similar microfluidic iMiDEV module at a different research center for the synthesis of [68Ga]Ga-FAPI-46 with limited runs. Our study demonstrated the potential of microfluidic methods for efficient and reliable radiometal-based radiopharmaceutical synthesis, contributing valuable insights for future advancements in this field and paving the way for routine clinical applications in the near future.


Twitter Analysis of the Nonmedical Use and Side Effects of Methylphenidate: Machine Learning Study.
Methylphenidate, a stimulant used to treat attention deficit hyperactivity disorder, has the potential to be used nonmedically, such as for studying and recreation. In an era when many people actively use social networking services, experience with the nonmedical use or side effects of methylphenidate might be shared on Twitter. The purpose of this study was to analyze tweets about the nonmedical use and side effects of methylphenidate using a machine learning approach. A total of 34,293 tweets mentioning methylphenidate from August 2018 to July 2019 were collected using searches for "methylphenidate" and its brand names. Tweets in a randomly selected training dataset (6860/34,293, 20.00%) were annotated as positive or negative for two dependent variables: nonmedical use and side effects. Features such as personal noun, nonmedical use terms, medical use terms, side effect terms, sentiment scores, and the presence of a URL were generated for supervised learning. Using the labeled training dataset and features, support vector machine (SVM) classifiers were built and the performance was evaluated using F1 scores. The classifiers were applied to the test dataset to determine the number of tweets about nonmedical use and side effects. Of the 6860 tweets in the training dataset, 5.19% (356/6860) and 5.52% (379/6860) were about nonmedical use and side effects, respectively. Performance of SVM classifiers for nonmedical use and side effects, expressed as F1 scores, were 0.547 (precision: 0.926, recall: 0.388, and accuracy: 0.967) and 0.733 (precision: 0.920, recall: 0.609, and accuracy: 0.976), respectively. In the test dataset, the SVM classifiers identified 361 tweets (1.32%) about nonmedical use and 519 tweets (1.89%) about side effects. The proportion of tweets about nonmedical use was highest in May 2019 (46/2624, 1.75%) and December 2018 (36/2041, 1.76%). The SVM classifiers that were built in this study were highly precise and accurate and will help to automatically identify the nonmedical use and side effects of methylphenidate using Twitter.


Coordination Chemistry of Sulfur-Containing Bifunctional Chelators: Toward in Vivo Stabilization of 64Cu PET Imaging Agents for Alzheimer's Disease.
The broader utilization of 64Cu positron emission tomography (PET) imaging agents has been hindered by the unproductive demetalation induced by bioreductants. To advance the development of 64Cu-based PET imaging tracers for Alzheimer's Disease (AD), there is a need for novel ligand design strategies. In this study, we developed sulfur-containing dithiapyridinophane (N2S2) bifunctional chelators (BFCs) as well as all nitrogen-based diazapyridinophane (N4) BFCs to compare their abilities to chelate Cu and target Aβ aggregates. Through spectrophotometric titrations and electrochemical measurements, we have demonstrated that the N2S2-based BFCs exhibit >10 orders of magnitude higher binding affinity toward Cu(I) compared to their N4-based counterparts, while both types of BFCs exhibit high stability constants toward Cu(II). Notably, solid state structures for both Cu(II) and Cu(I) complexes supported by the two ligand frameworks were obtained, providing molecular insights into their copper chelating abilities. Aβ binding experiments were conducted to study the structure-affinity relationship, and fluorescence microscopy imaging studies confirmed the selective labeling of the BFCs and their copper complexes. Furthermore, we investigated the potential of these ligands for the 64Cu-based PET imaging of AD through radiolabeling and autoradiography studies. We believe our findings provide molecular insights into the design of bifunctional Cu chelators that can effectively stabilize both Cu(II) and Cu(I) and, thus, can have significant implications for the development of 64Cu PET imaging as a diagnostic tool for AD.


Diagnosis and management of occult gastrointestinal bleeding: visualization of the small bowel lumen by fiberoptic colonoscope.
In 95 per cent of patients, the source of gastrointestinal bleeding can be determined by the usual roentgenographic and endoscopic methods. In the remaining five per cent of patients, the source of bleeding is obscure and special diagnostic procedures will be necessary. Among the special procedures that have been reported are the fluorescein string test, stepwise aspiration of intestinal contents, detection of radioactive red blood cells by stepwise aspiration, Geiger counter or scanning, and selective visceral angiography. An additional procedure, reported here, combines exploratory celiotomy with total endoscopic examination of the bowel.


Xeroderma pigmentosum complementation group C cells remove pyrimidine dimers selectively from the transcribed strand of active genes.
We have measured the removal of UV-induced pyrimidine dimers from DNA fragments of the adenosine deaminase (ADA) and dihydrofolate reductase (DHFR) genes in primary normal human and xeroderma pigmentosum complementation group C (XP-C) cells. Using strand-specific probes, we show that in normal cells, preferential repair of the 5' part of the ADA gene is due to the rapid and efficient repair of the transcribed strand. Within 8 h after irradiation with UV at 10 J m-2, 70% of the pyrimidine dimers in this strand are removed. The nontranscribed strand is repaired at a much slower rate, with 30% dimers removed after 8 h. Repair of the transcribed strand in XP-C cells occurs at a rate indistinguishable from that in normal cells, but the nontranscribed strand is not repaired significantly in these cells. Similar results were obtained for the DHFR gene. In the 3' part of the ADA gene, however, both normal and XP-C cells perform fast and efficient repair of either strand, which is likely to be caused by the presence of transcription units on both strands. The factor defective in XP-C cells is apparently involved in the processing of DNA damage in inactive parts of the genome, including nontranscribed strands of active genes. These findings have important implications for the understanding of the mechanism of UV-induced excision repair and mutagenesis in mammalian cells.


Kearns syndrome or Kearns disease. Further evidence of a genuine entity in a case with uncommon features.
A 20-year-old man with the characteristic findings of infantile onset Kearns syndrome is described. Morphological and biochemical investigations proved a mitochondrial disease which we believe to be the cause of the symptoms in various organs. We assume an autosomal-dominant inheritance, the marker sign of which is blepharoptosis in several family members. Characteristic clinical, morphological and biochemical findings, combined with an autosomal-dominant inheritance with very variable expression, mark the Kearns syndrome as an individual disease, not as a symptom complex (syndrome). Kearns disease can be divided into three forms--an infantile form ("Kearns-Sayre syndrome') with early onset, rapid progression, multisystemic involvement and a severe course; and a juvenile and an adult form with onset in the second, respectively third (or later) decades with a generally slower and more benign course and less widespread expression in various organ systems. Furthermore, the occurrence of a curious orthoptic abnormality is described, indicating one of the possible ways to avoid diplopia in chronic progressive external ophthalmoplegia: the coexistence of normal and gliding abnormal retinal correspondence.


Carcinoembryonic antigen for monitoring patients with small cell carcinoma of the lung during treatment.
Carcinoembryonic antigen (CEA) was measured at specific intervals in the plasma of 56 patients with small cell carcinoma of the lung. Of these patients, 47 had serial analyses for varying periods during their illness, 42 had pretreatment CEA levels, and 17 of the latter patients had determinations throughout the entire course of their disease. Pretreatment CEA levels were elevated above 2.5 ng/ml for 74% of the 42 patients and above 5.0 ng/ml for 48%. Although exceptions were noted, in general, a direct relationship was found between pretreatment CEA levels and extent of disease or tumor burden. Initial stage of disease, however, was more predictive of survival than was the pretreatment CEA level. With response to therapy, a corresponding decrease in CEA levels occurred for patients with an elevated pretreatment level. For those patients with a pretreatment CEA level below 5.0 ng/ml, an immediate slight increase in level was often seen associated with response and followed by a subsequent fall after one month. A rising CEA level was usually found with recurrence or progression of disease after initial response and occurred frequently prior to clinical evidence of progression. In combination with careful clinical evaluation, serial CEA measurements can aid in assessing tumor changes associated with treatment in patients with small cell carcinoma of the lung particularly at the times of recurrence or disease progression following a partial or complete response.


Decrease of ocular pressure with oral metyrapone. A double-masked crossover trial.
To examine the effect of a decrease in plasma cortisol level on intraocular pressure, 14 subjects with elevated ocular pressure were given oral metyrapone tartrate (an inhibitor of adrenal cortisol biosynthesis) and a placebo in a double-masked, two-period, crossover trial. Ocular pressure and the adrenal response were monitored over a seven-hour period. The decrease in ocular pressure after metyrapone administration was significantly greater than that after placebo. The 14 subjects could be designated as responders or nonresponders by decline of ocular pressure after metyrapone administration compared with response after placebo. The responders could be further differentiated from the nonresponders by a smaller decrease in ocular pressure over time in response to the placebo and by a smaller decrease over time of plasma cortisol level in response to metyrapone. These results provide further evidence that plasma glucocorticoids may play a role in the regulation of ocular pressure.


Double dissociation between hippocampal and parahippocampal responses to object-background context and scene novelty.
Several recent models of medial temporal lobe (MTL) function have proposed that the parahippocampal cortex processes context information, the perirhinal cortex processes item information, and the hippocampus binds together items and contexts. While evidence for a clear functional distinction between the perirhinal cortex and other regions within the MTL has been well supported, it has been less clear whether such a dissociation exists between the hippocampus and parahippocampal cortex. In the current study, we use a novel approach applying a functional magnetic resonance imaging adaptation paradigm to address these issues. During scanning, human subjects performed an incidental target detection task while viewing trial-unique sequentially presented pairs of natural scenes, each containing a single prominent object. We observed a striking double dissociation between the hippocampus and parahippocampal cortex, with the former showing a selective sensitivity to changes in the spatial relationship between objects and their background context and the latter engaged only by scene novelty. Our findings provide compelling support for the hypothesis that rapid item-context binding is a function of the hippocampus, rather than the parahippocampal cortex, with the former acting to detect relational novelty of this nature through its function as a match-mismatch detector.


Species replacement dominates megabenthos beta diversity in a remote seamount setting.
Seamounts are proposed to be hotspots of deep-sea biodiversity, a pattern potentially arising from increased productivity in a heterogeneous landscape leading to either high species co-existence or species turnover (beta diversity). However, studies on individual seamounts remain rare, hindering our understanding of the underlying causes of local changes in beta diversity. Here, we investigated processes behind beta diversity using ROV video, coupled with oceanographic and quantitative terrain parameters, over a depth gradient in Annan Seamount, Equatorial Atlantic. By applying recently developed beta diversity analyses, we identified ecologically unique sites and distinguished between two beta diversity processes: species replacement and changes in species richness. The total beta diversity was high with an index of 0.92 out of 1 and was dominated by species replacement (68%). Species replacement was affected by depth-related variables, including temperature and water mass in addition to the aspect and local elevation of the seabed. In contrast, changes in species richness component were affected only by the water mass. Water mass, along with substrate also affected differences in species abundance. This study identified, for the first time on seamount megabenthos, the different beta diversity components and drivers, which can contribute towards understanding and protecting regional deep-sea biodiversity.


Ionotropic Receptor IR75q.2 Mediates Avoidance Reaction to Nonanoic Acid in the Fall Armyworm Spodoptera frugiperda (Lepidoptera, Noctuidae).
Ionotropic receptors (IRs) play an important role in olfaction, but little is known in nondrosophila insects. Here, we report in vitro and in vivo functional characterization of IR75q.2 in the invasive moth pest Spodoptera frugiperda. First, 13 IRs (including four coreceptor IRs) were found specifically or highly expressed in adult antennae. Second, these IRs were tested for responding profiles to 59 odorants using the Xenopus oocyte expression system, showing that only SfruIR75q.2 responded to 8-10C fatty acids and their corresponding aldehydes, with SfruIR8a as the only coreceptor. Third, the three acids (especially nonanoic acid) showed repellent effects on moth's behavior and oviposition, but the repellence significantly reduced to the insects with IR75q.2 knockout by CRISPR/Cas9. Taken together, our study reveals the function of SfruIR75q.2 in perception of acid and aldehyde odorants and provides the first in vivo evidence for olfactory function of an odor-specific IR in Lepidoptera.


Self-expanding foam for prehospital treatment of intra-abdominal hemorrhage: 28-day survival and safety.
Intracavitary noncompressible hemorrhage remains a significant cause of preventable death on the battlefield and in the homeland. We previously demonstrated the hemostatic efficacy of an in situ self-expanding poly(urea)urethane foam in a severe, closed-cavity, hepatoportal exsanguination model in swine. We hypothesized that treatment with, and subsequent explantation of, foam would not adversely impact 28-day survival in swine. Following a closed-cavity splenic transection, animals received either fluid resuscitation alone (control group, n = 6) or resuscitation plus foam treatment at doses of 100 mL (n = 6), 120 mL (n = 6), and 150 mL (n = 2). Foam was allowed to polymerize in situ and was explanted after 3 hours. The animals were recovered and monitored for 28 days. All 18 animals in the 100-mL, 120-mL, and control groups survived to the 28-day endpoint without complications. The 150-mL group was terminated after the acute phase (n = 2). En bloc explantation of the foam took less than 2 minutes and was associated with millimeter-sized remnant particles. All foam animals required some level of enteric repair (imbrication or resection). Excluding the aborted 150-mL group, all animals survived, with no differences in renal or hepatic function, serum chemistries, or semiquantitative abdominal adhesion scores. Histologic analysis demonstrated that remnant particles were associated with a fibrotic capsule and mild inflammation, similar to that of standard suture reaction. In addition, safety testing (including genotoxicity, pyrogenicity, and cytotoxicity) was performed consistent with the ISO-10993 standard, and the materials passed all tests. For a distinct dose range, 28-day recovery after foam treatment and explantation for noncompressible, intra-abdominal hemorrhage is not associated with significant physiologic or biochemical evidence of end-organ dysfunction. A foam volume exceeding the maximum tolerable dose was identified. Bowel repair is required to ensure survival.


Regulation of asparagine synthetase gene expression by amino acid starvation.
We have studied the regulation of expression of the asparagine synthetase (AS) gene in ts11 cells, a mutant of BHK hamster cells which encodes a temperature-sensitive AS and therefore does not produce endogenous asparagine at 39.5 degrees C. Incubation of ts11 cells at the nonpermissive temperature drastically increases the level of AS mRNA, and the stimulation of AS mRNA expression is effectively suppressed by the addition of asparagine to the medium. We show here that regulation of AS gene expression involves cis-acting elements which are contained in the mRNA as well as in the 5' genomic region. When a plasmid containing the human AS cDNA under the control of the human AS promoter region was stably transfected into ts11 cells, the expression of human AS RNAs was regulated as that of the endogenous hamster transcripts, indicating that this construct contained all cis elements necessary for regulation. Expression of the AS cDNA in ts11 cells under the control of a constitutive foreign promoter was also regulated by the concentration of asparagine, and this regulation required translation. When we introduced by mutagenesis a number of stop codons in the AS cDNA, the mutant mRNAs with short open reading frames were expressed at low levels that were not increased by asparagine deprivation. Inhibition of protein and RNA synthesis also prevented down-regulation of AS mRNA levels by high concentrations of asparagine. In a parallel series of experiments, we showed that an AS DNA fragment including the promoter and first exon can also regulate RNA expression in response to asparagine concentration. Furthermore, similar increases in the levels of AS RNAs are produced not only by asparagine deprivation in ts11 cells but also by deprivation of human and wild-type BHK cells of leucine, isoleucine, or glutamine. Thus, regulation of AS gene expression is a response to amino acid starvation through mechanisms which appear to involve both changes in RNA stability and change in the rates of transcription initiation or elongation.


Stem cell-derived models of spinal neurulation.
Neurulation is a critical step in early embryonic development, giving rise to the neural tube, the primordium of the central nervous system in amniotes. Understanding this complex, multi-scale, multi-tissue morphogenetic process is essential to provide insights into normal development and the etiology of neural tube defects. Innovations in tissue engineering have fostered the generation of pluripotent stem cell-based in vitro models, including organoids, that are emerging as unique tools for delving into neurulation mechanisms, especially in the context of human development. Each model captures specific aspects of neural tube morphogenesis, from epithelialization to neural tissue elongation, folding and cavitation. In particular, the recent models of human and mouse trunk morphogenesis, such as gastruloids, that form a spinal neural plate-like or neural tube-like structure are opening new avenues to study normal and pathological neurulation. Here, we review the morphogenetic events generating the neural tube in the mammalian embryo and questions that remain unanswered. We discuss the advantages and limitations of existing in vitro models of neurulation and possible future technical developments.


[The tissue specificity of the protective action of cytoplasmic factors on the membrane-bound system of Ca2+ transport in the sarcoplasmic reticulum of the heart and the skeletal muscles].
Whether soluble cytoplasmic factors (SCF) can protect the membrane-associated Ca2+ transport system in sarcoplasmic reticulum (SR) was studied in controls, stress and adaptation to it. Ca-transport was damaged in stress much more in the skeletal muscles compared to the heart. Initial performance of Ca-pump in the skeletal muscles was decreased by 43%, in the heart--no change. After stress, thermoresistance of the enzyme in the skeletal muscles was 3-5 times less than in the myocardium. The enzyme reaction on adaptation and stress in both tissues is the same--a significant rise in Ca-pump thermoresistance. When preadapted animals are exposed to stress, deceleration of the initial speed and resistance of the enzyme in the myocardium are fully prevented while in the skeletal muscles Ca-pump resistance is higher than in the controls. In the heart, SCF have a protective effect on thermoresistance. In the skeletal muscles, SCF protect initial speed of the enzyme in the control, in stress and adaptation to it. A cross-over protective SCF (from adapted animals) effect was found on the stress-impaired membrane system of Ca-transport. Adapted SCF elevate 2.5-fold thermoresistance of the enzyme. Correlation between cytoplasmic and membrane protective mechanisms in stress and adaptation to it is considered.


Reclaimed water driven lettuce cultivation in a hydroponic system: the need of micropollutant removal by advanced wastewater treatment.
For a novel approach of resource-efficient water reuse, a municipal wastewater treatment plant was extended at pilot scale for advanced wastewater treatment, i.e., ozonation and biological activated carbon filtration, and a hydroponic system for reclaimed water driven lettuce cultivation. The treatment specific wastewater lines with the corresponding lettuce plants, differentiated into roots and shoots, were monitored for priority wastewater micropollutants, i.e., acesulfame (sweetener), caffeine (stimulant), carbamazepine, diclofenac, ibuprofen, sulfamethoxazole with acetyl-sulfamethoxazole (human pharmaceuticals), 1H-benzotriazole, and 4/5-methylbenzotriazole (industrial chemicals). As clearly demonstrated, conventional tertiary treatment could not efficiently clean up wastewater. Removal efficiencies ranged from 3% for carbamazepine to 100% for ibuprofen. The resulting pollution of the hydroponic water lines led to the accumulation of acesulfame, carbamazepine, and diclofenac in lettuce root systems at 32.0, 69.5, and 135 μg kg-1 and in the uptake of acesulfame and carbamazepine into lettuce shoots at 23.4 and 120 μg kg-1 dry weight, respectively. In contrast, both advanced treatment technologies when operating under optimized conditions achieved removal efficiencies of > 90% also for persistent micropollutants. Minimizing the pollution of reclaimed water thus met one relevant need for hydroponic lettuce cultivation.


Simulations of the Ultra-Fast Kinetics in Ni-Si-C Ternary Systems under Laser Irradiation.
We present a method for the simulation of the kinetic evolution in the sub µs timescale for composite materials containing regions occupied by alloys, compounds, and mixtures belonging to the Ni-Si-C ternary system. Pulsed laser irradiation (pulses of the order of 100 ns) promotes this evolution. The simulation approach is formulated in the framework of the phase-field theory and it consists of a system of coupled non-linear partial differential equations (PDEs), which considers as variables the following fields: the laser electro-magnetic field, the temperature, the phase-field and the material (Ni, Si, C, C clusters and Ni-silicides) densities. The model integrates a large set of materials and reaction parameters which could also self-consistently depend on the model variables. A parameter calibration is also proposed, specifically suited for the wavelength of a widely used class of excimer lasers (λ = 308 nm). The model is implemented on a proprietary laser annealing technology computer-aided design (TCAD) tool based on the finite element method (FEM). This integration allows, in principle, numerical solutions in systems of any dimension. Here we discuss the complex simulation trend in the one-dimensional case, considering as a starting state, thin films on 4H-SiC substrates, i.e., a configuration reproducing a technologically relevant case study. Simulations as a function of the laser energy density show an articulated scenario, also induced by the variables' dependency of the materials' parameters, for the non-melting, partial-melting and full-melting process conditions. The simulation results are validated by post-process experimental analyses of the microstructure and composition of the irradiated samples.


Cell cycle perturbations in acute myeloid leukemia samples following in vitro exposures to therapeutic agents.
Cell cycle checkpoints establish the timing and strength of arrest, repair and apoptosis responses to damaging treatments. We designed flow cytometric assays to measure cell cycle arrest and apoptosis in acute myeloid leukemia (AML) samples treated in vitro with relevant therapeutic agents so as to functionally characterize checkpoints in these samples and to ask if checkpoint abnormalities are common in AML and contribute to therapeutic failures. We show here that cell cycle responses to daunomycin (DNR), cytosine arabinoside (ARA-C) and gamma irradiation (RAD) were reproducibly treatment agent- and dose-dependent and distinct in different myeloid cell lines. DNR treatments differentially induced cell accumulations in the gap 2 and mitosis (G2/M) phases of the cell cycle and/or in the gap 1 (G1) phase, as did RAD, while ARA-C induced accumulations in the DNA synthesis (S) phase or in the G1 phase. Flow cytometric gates were devised to exclude lymphocytes and mature neutrophils in analyses of primary myeloid cell samples. Cell subsets in bone marrow samples from normal donors were thus enriched for myeloid constituents and used as normal myeloid cell controls. Proliferating cell nuclear antigen (PCNA) immunostaining was used to further identify actively dividing cell subpopulations in primary cell samples. AML samples were similarly analyzed and the majority showed lower DNA synthesis cell cycle phase (S) fractions and lower PCNA-positive fractions than normal myeloid cells, suggesting that AMLs are generally less proliferative in these culture conditions. Exceptional AML samples with high S phase fractions had cytogenetic abnormalities associated with poor prognosis. Most AML samples mounted weak cell cycle responses relative to normal myeloid cells, while a minority showed robust, agent-specific cell cycle arrests. This non-responsiveness was not simply associated with lower cycling indices-neither the response patterns nor the degrees of response were correlated with untreated S phase fractions or with PCNA-positive fractions. Cell cycle responses were also not associated with clinical parameters including patient age, FAB class, or white blood cell count, nor with immunophenotypic features including CD34 status, nor with specific cytogenetic markers. This suggests that functional cell cycle response assays could provide unique diagnostic information in AML. These assays might also have prognostic value as ARA-C induced G1 arrests and DNR-specific G2/M arrests tended to be associated with failure to achieve clinical remission. In addition, G1 arrests after ARA-C and G2/M accumulations after DNR treatments tended to be more robust in samples that had previously been shown to be more highly immunopositive for bcl-2 expression. This data suggests that the association of bcl-2 expression with particular cell cycle responses to therapeutic agents may contribute to the association of bcl-2 with poor clinical responses in AML. These data provide the basis for further laboratory studies aimed at examining specific cell cycle arrests as mechanisms of therapeutic resistance and prospective studies aimed at rigorously assessing the prognostic utility of in vitro assays of checkpoint function.


Human intestinal cell lines as in vitro tools for electrolyte transport studies with relevance to secretory diarrhoea.
Intestinal cells from human colorectal adenocarcinomas have been established in culture as monolayers of homogeneous polarised cells on permeable filters with various degrees of cell differentiation. These cell lines allow to avoid some of the difficulties of studying water and electrolyte transport through whole intestinal tissue or isolated intestinal cells. Cell lines, such as Caco-2, T-84, HT-29, HRT-18 which exhibit electrogenic chloride secretion induced by various hormones, neurotransmitters, enterotoxins, and other chemical compounds, can be used to study basic biochemical mechanism and their interrelation in the process of chloride secretion which is associated with many diarrhoeal diseases. Identification and use of cell lines having absorptive properties will equally contribute to the understanding of absorptive processes, their regulation and biochemical basis. Cell lines in which both secretory and absorptive processes coexist are valuable for understanding the complex interrelation of the two principal processes of transport mechanism including its biochemical and molecular correlates. Intestinal cell lines can also be used for screening efficacies of potential secretagogues and antisecretagogues.


A novel injectable in situ forming gel based on carboxymethyl hexanoyl chitosan/hyaluronic acid polymer blending for sustained release of berberine.
An in situ forming gel based on simply blending carboxymethyl hexanoyl chitosan (CHC) with low molecular weight hyaluronic acid (LMW HA) was developed, without needing cross-linking, photopolymerization or thermal treatments. The CHC/LMW HA blends formed nanoparticles and then rapidly transformed into supermolecular hydrogels under stirring. The gel formation mechanism was examined by Förster resonance energy transfer (FRET). The gels were injectable, cytocompatible and biodegradable, and showed shape-persistent behavior and adhesive property. Berberine, an anti-apoptotic and anti-arthritis naturally occurring compound, was encapsulated within the CHC/LMW HA gels. The gels demonstrated a pH-responsive characteristic which were able to release berberine in a sustained manner at pH 6.0 (simulating inflamed arthritic articular cartilage) and the degradation rates were accelerated at pH 7.4 (simulating healed normal tissue). The berberine-loaded gels effectively protected chondrocytes against sodium nitroprusside-induced apoptosis. The gels may be potentially useful as an injectable system for intra-articular drug delivery and cartilage tissue engineering.


[Use of thymalin in the complex treatment of patients with cancer of the uterus].
The results of a clinico-immunologic study of thymalin as a component of complex antitumor treatment of 21 cases of cancer of corpus uteri pointed to a pronounced immunomodulating action of the drug on T-cell-mediated immunity. It was inferred that thymalin should be used for immunocorrection in complex treatment for the said neoplasms provided T-cell-mediated immunity is monitored.


Prone positioning as a bridge to recovery from refractory hypoxaemia following lung transplantation.
Refractory hypoxaemia is the leading cause of mortality in the postoperative period after lung transplantation. The role of prone positioning as a rescue therapy in this setting has not been assessed. We evaluated its effects in lung transplant recipients presenting refractory hypoxaemia following the surgery. Prospectively collected data from 131 consecutive adult patients undergoing lung transplantation between January 2013 and December 2014 were evaluated. Twenty-two patients received prone position therapy. Indications, associated complications, time to initiation and duration of the manoeuvre were analysed and the effects of prone position on gas exchange were evaluated. Finally, outcomes in this cohort were compared against the rest of lung transplant recipients. Prone positioning was more frequently implemented within the first 72 h (68.2%) and its main indication was primary graft dysfunction. The manoeuvre was maintained during a median of 21 h. After prone position, the pressure of arterial oxygen/fraction of inspired oxygen ratio significantly increased from 81.0 mmHg [interquartile range (IQR) 71.5-104.0] to 220.0 (IQR 160.0-288.0) (P < 0.001). No complications related with the technique were reported. Patients who underwent the manoeuvre had longer hospital stay [50.0 days (IQR 36.0-67.0) vs 30.0 (IQR 23.0-56.0), P = 0.006] than the rest of the population. No differences were found comparing either 1-year mortality (9.1% vs 15.6%; P = 0.740) or 1-year graft function [forced expiratory volume in 1 second of 70.0 (IQR 53.0-83.0) vs 68.0 (IQR 53.5-80.5), P = 0.469]. Prone positioning is safe and significantly improves gas exchange in patients with refractory hypoxaemia after lung transplantation. It should be considered as a possible treatment in these patients.