Dynamics of photosystem II: a proteomic approach to thylakoid protein complexes.
Oxygenic photosynthesis produces various radicals and active oxygen species with harmful effects on photosystem II (PSII). Such photodamage occurs at all light intensities. Damaged PSII centres, however, do not usually accumulate in the thylakoid membrane due to a rapid and efficient repair mechanism. The excellent design of PSII gives protection to most of the protein components and the damage is most often targeted only to the reaction centre D1 protein. Repair of PSII via turnover of the damaged protein subunits is a complex process involving (i) highly regulated reversible phosphorylation of several PSII core subunits, (ii) monomerization and migration of the PSII core from the grana to the stroma lamellae, (iii) partial disassembly of the PSII core monomer, (iv) highly specific proteolysis of the damaged proteins, and finally (v) a multi-step replacement of the damaged proteins with de novo synthesized copies followed by (vi) the reassembly, dimerization, and photoactivation of the PSII complexes. These processes will shortly be reviewed paying particular attention to the damage, turnover, and assembly of the PSII complex in grana and stroma thylakoids during the photoinhibition-repair cycle of PSII. Moreover, a two-dimensional Blue-native gel map of thylakoid membrane protein complexes, and their modification in the grana and stroma lamellae during a high-light treatment, is presented.


Integrated Differential Regulatory Analysis Reveals a Novel Prognostic 36-Gene Signature for Gastric Cancer in Asian Population.
Gastric cancer is one of the most common cancers and has very high incidence and mortality rate in Asian population. To tackle the problems of infiltration and heterogeneity, more accurate biomarkers for diagnosis and prognosis as well as effective targets for treatment are needed to achieve better outcomes of gastric cancer patients. Recently, methods and algorithms for analyzing high-throughput sequencing data have greatly facilitated the molecular profiling of gastric cancer. Nevertheless, prognostic biomarkers for gastric cancer that can be potentially applied in clinic are still lacking. In this study, we performed differential regulatory analysis based on gene co-expression network for four different cohorts of Asian gastric cancer samples and their clinical data. We identified a 36-gene prognostic signature specific for gastric cancer, particularly for Asian population. We further analyzed differential regulatory patterns related to these featured genes, such as C1S, and suggested hypotheses for investigating their roles in gastric cancer pathogenesis. Findings from present study suggest a 36-gene signature which is based on differential regulatory analysis and can predict the prognosis of gastric cancer. Our research explores molecular mechanism of gastric cancer at transcriptional regulation level and provides potential drug targets. This integrated biomarker searching scheme is extendable to other cancer study for not only prognostic prediction, but also pathogenesis.


[Tobacco--a highly efficient producer of vaccines].
Along with the depreciation of tobacco as a source of nicotine-containing commercial products, the increase of its appreciation as a potential producer of recombinant therapeutical proteins can be observed. Two species of tobacco--Nicotiana tabacum L. and N. benthamiana are easily grown by well established methods of field or green-house cultivation or cell culture, yield high biomass and soluble protein content, can be easily transformed by several methods and are not food for humans or feed for animals. Expression of foreign proteins, including vaccines, can be achieved in those plants either through stable transformation of nuclear or plastid (chloroplast) genomes or by transient transformation using infection with plant virus or bacteria--Agrobacterium tumefaciens (agroinfiltration). The most advanced mode of agrofiltration termed magnifection, which combines benefits of virus and Agrobacterium and depends on using Agrobacterium with viral pro-vectors, enables high-yield and rapid expression of therapeutical proteins, even in a few days, and can be employed on an industrial scale. Expression of many antigenic proteins, which may serve as antiviral, antibacterial, antiprotozoan and anticancer vaccines, and additionally a few autoantigens designed for the treatment of autoimunogenic diseases, like diabetes, have been achieved in tobacco. To date, a vaccine against Newcastle virus disease in poultry produced by tobacco cell culture has been approved for commercial application and several other vaccines are in advanced stage of development. The possibility of a high-level production of vaccines in tobacco against pandemic influenza or anthrax and plague due to a bioterroristic attack, as well as of individualised anticancer vaccines against non-Hodgkin's lymphoma (NHL) in a much shorter period of time than by traditional methods became realistic and hence caused increased interest in tobacco as a high-efficient producer of vaccines not only of specialistic biotechnology firms but also a big pharmaceutical corporation and a department of defence.


alpha-Aminoadipate pathway for the biosynthesis of lysine in lower eukaryotes.
Bacteria and green plants use the diaminopimelate pathway for the biosynthesis of the essential amino acid, lysine; however, yeast and other higher fungi use the alpha-aminoadipate (AA) pathway. The AA pathway has been investigated in detail biochemically, genetically, and in terms of regulatory mechanisms in the baker's yeast Saccharomyces cerevisiae. The genetic analysis of lysine auxotrophs of S. cerevisiae revealed that there are more than 12 lysine genes for 8 enzyme-catalyzed steps. Lysine genes are not linked to each other and seven of the genes are mapped on six different linkage groups (chromosomes). The gene-enzyme relationships have been determined for ten of the lysine loci which include two unlinked gene functions required for each of AA reductase (LYS2 and LYS5) and Saccharopine reductase (LYS9 and LYS14). Five of the lysine enzymes are localized in mitochondria and three in cytosol. The lysine pathway of S. cerevisiae is regulated by feedback inhibition and end product repression. Two, and possibly three, of the enzymes exhibit general control of amino acid biosynthesis and at least five of the enzymes coded for, by unlinked genes, are simultaneously depressed in a regulatory (repressor) gene-mutant.


Direct phenotypic analysis of human MHC class I antigen presentation: visualization, quantitation, and in situ detection of human viral epitopes using peptide-specific, MHC-restricted human recombinant antibodies.
The advent in recent years of the application of tetrameric arrays of class I peptide-MHC complexes now enables us to detect and study rare populations of Ag-specific CD8(+) T cells. However, available methods cannot visualize or determine the number and distribution of these TCR ligands on individual cells nor detect APCs in tissues. In this study, we describe for the first time studies of human class I peptide-MHC ligand presentation. These studies were facilitated by applying novel tools in the form of peptide-specific, HLA-A2-restricted human recombinant Abs directed toward a viral epitope derived from human T cell lymphotropic virus type I. Using a large human Ab phage display library, we isolated a large panel of recombinant Fab Abs that are specific for a particular peptide-MHC class I complex in a peptide-dependent, MHC-restricted manner. We used these Abs to visualize the specific complex on APCs and virus-infected cells by flow cytometry, to quantify the number of, and visualize in situ, a particular complex on the surface of APCs bearing complexes formed by naturally occurring active intracellular processing of the cognate viral Ag. These findings demonstrate our ability to transform the unique fine specificity, but low intrinsic affinity of TCRs into high affinity soluble Ab molecules endowed with a TCR-like specificity toward human viral epitopes. These molecules may prove to be crucial useful tools for studying MHC class I Ag presentation in health and disease as well as for therapeutic purposes in cancer, infectious diseases, and autoimmune disorders.


Preclinical Evaluation and First Patient Application of 99mTc-PSMA-I&S for SPECT Imaging and Radioguided Surgery in Prostate Cancer.
Initial studies in patients have demonstrated the suitability of 111In-PSMA-I&T (111In-DOTAGA-(3-iodo-y)-f-k-Sub(KuE)) (PSMA is prostate-specific membrane antigen and I&T is imaging and therapy) for radioguided surgery (RGS) of small metastatic prostate cancer (PCa) soft-tissue lesions. To meet the clinical need for a more cost-effective alternative, the PSMA-I&T-based tracer concept was adapted to 99mTc-labeling chemistry. Two PSMA-I&T-derived inhibitors with all-L-serine- (MAS3) and all-D-serine- (mas3) chelating moieties were evaluated in parallel, and a kit procedure for routine 99mTc labeling was developed. PSMA affinities (IC50) and internalization kinetics of 99mTc-MAS3-y-nal-k(Sub-KuE) and 99mTc-mas3-y-nal-k(Sub-KuE) (99mTc-PSMA-I&S for imaging and surgery) were determined using LNCaP cells and (125I-BA)KuE as a radioligand and reference standard. In vivo metabolite analyses and biodistribution studies were performed using CD-1 nu/nu and LNCaP tumor-bearing CB-17 severe combined immunodeficiency mice. The pharmacokinetics of 99mTc-PSMA-I&S in humans were investigated in a patient with advanced metastatic PCa via sequential planar whole-body SPECT imaging at 1, 3, 5, and 21 h after injection. Additionally, preoperative SPECT/CT (12 h after injection) and 99mTc-PSMA-I&S-supported RGS (16 h after injection) were performed in 1 PCa patient with proven iliac and inguinal lymph node metastases. A robust and reliable kit-labeling procedure was established, allowing the preparation of 99mTc-MAS3-y-nal-k(Sub-KuE) and 99mTc-PSMA-I&S in consistently high radiochemical yield and purity (≥98%, n > 50 preparations). Because of its improved internalization efficiency and superior in vivo stability, 99mTc-PSMA-I&S was selected for further in vivo evaluation. Compared with 111In-PSMA-I&T, 99mTc-PSMA-I&S showed delayed clearance kinetics but identical uptake in PSMA-positive tissues in the LNCaP xenograft model (1 h after injection). In exemplary PCa patients, a relatively slow whole-body clearance of 99mTc-PSMA-I&S was observed due to high plasma protein binding (94%) of the tracer. This, however, promoted efficient tracer uptake in PCa lesions over time and led to steadily increasing lesion-to-background ratios up to 21 h after injection. Preoperative SPECT/CT showed a high 99mTc-PSMA-I&S uptake in all suspect lesions identified in previous 68Ga-HBED-CC-Ahx-KuE (68Ga-HBED-CC-PSMA) PET/CT, allowing for their successful intraoperative detection and resection during first-in-human RGS. Because of a straightforward and reliable kit production, 99mTc-PSMA-I&S represents a cost-effective, readily available alternative to 111In-PSMA-I&T. Initial patient data indicate its comparable or even superior performance as a probe for PSMA-targeted RGS and also hint toward the unexpected potential of 99mTc-PSMA-I&S as a SPECT imaging agent.


Differential Regulation of the Autophagy and Proteasome Pathways in Skeletal Muscles in Sepsis.
Skeletal muscle fiber atrophy develops in response to severe sepsis, but it is unclear as to how the proteolytic pathways that are involved in its development are differentially regulated. We investigated the link between sepsis-induced fiber atrophy and activation of the proteasome and autophagy pathways and whether the degree of activation is more severe and sustained in limb muscles than it is in the diaphragm. Randomized controlled experiment. Animal research laboratory. Adult male C57/BL6 mice. Two groups of animals were studied. The sepsis group was subjected to a cecal ligation and perforation technique, whereas the control (sham) group was subjected to abdominal surgery without cecal ligation and perforation. Measurements for both groups were performed 24, 48, and 96 hours after the surgical procedure. Atrophy was quantified in the diaphragm and tibialis anterior by measuring fiber diameter. Autophagy was evaluated using electron microscopic detection of autophagosomes and by measuring LC3B protein lipidation and autophagy-related protein expressions. Proteasomal degradation was quantified by measuring chymotrypsin-like activity of the 26S proteasome and messenger RNA expressions of muscle-specific E3 ligases. Sepsis triggered transient fiber atrophy in the diaphragm that lasted for 24 hours and prolonged atrophy in the tibialis anterior that persisted for 96 hours. The autophagy and proteasome pathways were activated in both muscles at varying intensities over the time course of sepsis. Activation was more pronounced in the tibialis anterior than in the diaphragm. Sepsis inhibited the V-Akt thymoma viral oncogene homolog 1 and complex 1 of the mammalian target of rapamycin pathways and stimulated the AMP-activated protein kinase pathway in both muscles. Sepsis triggers more severe and sustained muscle fiber atrophy in limb muscles when compared with respiratory muscle. This response is associated with enhanced proteasomal and autophagic proteolytic pathway activities and is triggered by inhibition of the AKT and complex 1 of the mammalian target of rapamycin pathways and activation of the AMPK pathway.


Exploring spatio-temporal variations in environmental impacts from eating out in the United Kingdom.
In this study we estimated the environmental burden of eating away from home based on emission factors of food recipes consumed in different regions and countries within the United Kingdom. Food based emissions were expressed in kg CO2 equivalent per capita per week and were calculated based on food consumption data between the years 2001 and 2018. Time series analysis was used to estimate emissions for the years 2019 and 2020 for all study areas. These results were used to estimate the endpoint impacts on human health as well as terrestrial and aquatic species during the study period. Finally, an estimate of the emissions for 2020 was also carried out based on available market data for the first 11 months of the year. This was subsequently compared with the forecasts calculated earlier to observe the impacts of Covid-19 led lockdowns on eating out and hence the emissions. By taking a subnational approach, we aimed to highlight the importance of appreciating similarities and differences among these regions and policy implications thereof. To the best knowledge of the authors this is the first and only study focusing on regional food-based emissions from eating out in the United Kingdom.


Non-invasive determination of port wine stain anatomy and physiology for optimal laser treatment strategies.
The treatment of port wine stains (PWSs) using a flashlamp-pumped pulsed dye laser is often performed using virtually identical irradiation parameters. Although encouraging clinical results have been reported, we propose that lasers will only reach their full potential provided treatment parameters match individual PWS anatomy and physiology. The purpose of this paper is to review the progress made on the technical development and clinical implementation of (i) infrared tomography (IRT), optical reflectance spectroscopy (ORS) and optical low-coherence reflectometry (OLCR) to obtain in vivo diagnostic data on individual PWS anatomy and physiology and (ii) models of light and heat propagation, predicting irreversible vascular injury in human skin, to select optimal laser wavelength, pulse duration, spot size and radiant exposure for complete PWS blanching in the fewest possible treatment sessions. Although non-invasive optical sensing techniques may provide significant diagnostic data, development of a realistic model will require a better understanding of relevant mechanisms for irreversible vascular injury.


Prophylactic surgical ligation of patent ductus arteriosus for prevention of mortality and morbidity in extremely low birth weight infants.
Patent ductus arteriosus (PDA) is associated with increased mortality and morbidity in preterm infants. Prophylactic indomethacin results in favorable intermediate outcomes such as reduction of significant PDA, need for surgical ligation, severe intraventricular hemorrhage and serious pulmonary hemorrhage without modifying long-term neurosensory outcomes. Little is known about the effectiveness and safety of prophylactic surgical closure of the PDA in extremely low birth weight (ELBW) infants. To identify and summarize evidence from randomized controlled trials investigating the effectiveness and safety of prophylactic surgical ligation of the PDA on mortality and morbidities of preterm infants less than 1000 g at birth as compared to no prophylaxis or prophylactic cyclooxygenase inhibitors. The standard search strategy for the Cochrane Neonatal Review Group was performed by two review authors. Searches were made of MEDLINE (1966 to December 2006), EMBASE (1980 to December 2006), the Cochrane Central Register of Controlled Trials (The Cochrane Library, Issue 4, 2006), and abstracts of annual meetings of the Society for Pediatric Research (1995 - 2006). Contacts were made with the authors of published articles. Randomized or quazi-randomized controlled trials that enrolled infants less than 28 weeks gestation or less than 1000 g at birth who are on assisted ventilation and/or supplemental oxygen without clinical signs of hemodynamic significance of the ductus arteriosus were considered. Trials addressing prophylactic surgical ligation of the patent ducts arteriosus (i.e. procedure done during the first 72 hours of life) versus no intervention or cyclooxygenase inhibitor prophylaxis were included. The primary outcome was bronchopulmonary dysplasia (BPD). Other important short and long-term neonatal outcomes were also considered. Standard methods of the Cochrane Collaboration and its Neonatal Review Group were used to assess the methodologic quality of the trials. Retrieved articles were assessed for eligibility and data was abstracted independently by two reviewer authors. Where data were incomplete, the primary investigators were contacted for further information and clarifications. For dichotomous outcomes, relative risk (RR) and its associated confidence interval were calculated. For continuous outcomes, treatment effect was expressed as mean difference and its calculated standard deviation. When appropriate, meta-analysis of pooled data was performed assuming a fixed effect model Only one eligible study that enrolled 84 ELBW infants was identified. The prophylactic group had ductal ligation performed within 24 hours of life following a pre-specified protocol, while the control group received standard care without indomethacin. Prophylactic surgical ligation of the PDA resulted in a statistically significant reduction of severe stage II or III necrotizing enterocolitis (NEC), [RR 0.25, 95% CI (0.08,0.83), p value 0.02, NNT 5]. The study found no statistically significant difference in mortality, severe grade III and IV intraventricular hemorrhage (IVH), BPD, and retinopathy of prematurity (ROP). Of note, the study was unblinded and underpowered to detect clinically important differences in the above mentioned outcomes. Prophylactic surgical ligation of the PDA did not decrease mortality or BPD in ELBW infants. A significant reduction of stage II or III NEC was noted. Based on the current evidence, the high rate of spontaneous closure, availability of effective safe medical therapies, and the potential short and long-term complications of surgical ligation, the use such prophylactic surgical therapy is not indicated in the management of the preterm infants.


Total heart replacement with dual centrifugal ventricular assist devices.
In an ovine feasibility study, we implanted two HeartMate-III centrifugal ventricular assist devices (VADs) for total heart replacement. With cardiopulmonary bypass support, both ventricles were transected at the atrioventricular groove, preserving a rim of ventricular tissue. The atrioventricular valves were excised, and the aorta and pulmonary artery were transected above the ventriculoarterial valves. An interatrial septal window was created by excising the foramen ovale. The VADs' sewing rings were attached to the left and right ventricular remnants, respectively. Outflow grafts were anastomosed to the aorta and pulmonary artery. The left VAD operated continuously at 4,500 rpm. Right VAD speed increased from 2,000 to 4,500 rpm in 500 rpm increments. Outflow graft flow, pressure, oxygen saturation, and shunt direction were recorded. The pulmonary artery to aortic ratio of flow and pressure increased from 0.26 and 0.15 (at 2,000 rpm) to 1.21 and 0.53, respectively (at 4,500 rpm). The interatrial shunt, which was right to left at lower right VAD speeds, progressed to bidirectional, then to left dominant as right VAD speed increased. Outflow-graft oxygen saturation was reflective of the shunt direction. In this acute experiment, total heart replacement with continuous flow VADs satisfactorily balanced left and right ventricular flows and preserved the physiologic circulatory response.


Housing First Impact on Costs and Associated Cost Offsets: A Review of the Literature.
Housing First (HF) programs for people who are chronically or episodically homeless, combining rapid access to permanent housing with community-based, integrated treatment, rehabilitation and support services, are rapidly expanding in North America and Europe. Overall costs of services use by homeless people can be considerable, suggesting the potential for significant cost offsets with HF programs. Our purpose was to provide an updated literature review, from 2007 to the present, focusing specifically on the cost offsets of HF programs. A systematic review was performed on MEDLINE and PsycINFO as well as Google and the Homeless Hub for grey literature. Study characteristics and key findings were extracted from identified studies. Where available, impact on service cost associated with HF (increase or decrease) and net impact on overall costs, taking into account the cost of HF intervention, were noted. Twelve published studies (4 randomized studies and 8 quasi-experimental) and 22 unpublished studies were retained. Shelter and emergency department costs decreased with HF, while impacts on hospitalization and justice costs are more ambiguous. Studies using a pre-post design reported a net decrease in overall costs with HF. In contrast, experimental studies reported a net increase in overall costs with HF. While our review casts doubt on whether HF programs can be expected to pay for themselves, the certainty of significant cost offsets, combined with their benefits for participants, means that they represent a more efficient allocation of resources than traditional services.


High-quality DNA sequence capture of 524 disease candidate genes.
The accurate and complete selection of candidate genomic regions from a DNA sample before sequencing is critical in molecular diagnostics. Several recently developed technologies await substantial improvements in performance, cost, and multiplex sample processing. Here we present the utility of long padlock probes (LPPs) for targeted exon capture followed by array-based sequencing. We found that on average 92% of 5,471 exons from 524 nuclear-encoded mitochondrial genes were successfully amplified from genomic DNA from 63 individuals. Only 144 exons did not amplify in any sample due to high GC content. One LPP was sufficient to capture sequences from <100-500 bp in length and only a single-tube capture reaction and one microarray was required per sample. Our approach was highly reproducible and quick (<8 h) and detected DNA variants at high accuracy (false discovery rate 1%, false negative rate 3%) on the basis of known sample SNPs and Sanger sequence verification. In a patient with clinical and biochemical presentation of ornithine transcarbamylase (OTC) deficiency, we identified copy-number differences in the OTC gene at exon-level resolution. This shows the ability of LPPs to accurately preserve a sample's genome information and provides a cost-effective strategy to identify both single nucleotide changes and structural variants in targeted resequencing.


Antibodies against MERS coronavirus in dromedary camels, Kenya, 1992-2013.
Dromedary camels are a putative source for human infections with Middle East respiratory syndrome coronavirus. We showed that camels sampled in different regions in Kenya during 1992-2013 have antibodies against this virus. High densities of camel populations correlated with increased seropositivity and might be a factor in predicting long-term virus maintenance.


Histopathological and immunohistochemical study of lambs experimentally infected with Fasciola hepatica and Schistosoma bovis.
The aim of this study was to investigate the cross-resistance between Fasciola hepatica and Schistosoma bovis in lambs assessing parasitologic, gross pathologic, histopathologic and immunohistochemical changes in liver and small intestine. Thirty Castellana breed lambs were divided into five comparable groups and exposed to F. hepatical S. bovis (group F/S), S. bovis/F. hepatica (group S/F), S. bovis (group S) or F. hepatica (group F) and six unexposed lambs were used as non-infected controls (group C). Primary patent infection with F. hepatica induced a lower number of schistosome eggs and a higher number of lymphocytes in intestinal and liver schistosome egg-induced granulomas in group F/S than in the groups S/F and S, liver damage being mainly attributed to F. hepatica. S. bovis infection followed by challenge with F. hepatica particularly increased the severity of the most significant liver alterations (cholangiohepatitis by F. hepatica and mesoendophlebitis by S. bovis) and F. hepatica seemed not to have an influence on established S. bovis infection. In addition, immunohistochemical results suggested that the predominant local immune response in both double-infected groups was different, being mainly a cell-mediated immune response in group F/S and a mucosal response in group S/F.


Multidisciplinary palliative treatment including isolated thoracic perfusion for progressive malignant pleural mesothelioma: a retrospective observational study.
To investigate the relative importance of isolated thoracic perfusion (ITP) in the multidisciplinary palliative treatment of progressive malignant pleural mesothelioma (MPM) patients. Fifty-two MPM patients with progressive disease after systemic chemotherapy with cisplatin and pemetrexed were submitted to 112 ITP using mitomycin C (25 mg/m2) and cisplatin (70 mg/m2) between 2000 and 2017. Isolation of the chest was achieved by insertion of stop-flow balloon catheters via femoral or iliac access. Primary endpoints were adverse events, tumor response rate, progression-free survival (PFS) and overall survival (OS) from initial ITP. Median interval-time from MPM diagnosis was 9 months. There were no perfusion-related postoperative deaths. The main procedure-related complication was persistent leakage of lymphatic fluid from the incision in less than 10% of ITP. No severe perfusion-related toxicity was reported, with grade 3 haematological toxicity and platinum-induced neurotoxicity in less than 8% of the patients. Following initial ITP, overall tumor response rate was 25%, median PFS was 7 months (IQR 5-10.5), and median OS was 16 months (IQR 12.5-21). After the last ITP, 14 patients received further therapies, including targeted therapy with cetuximab or bevacizumab. Non-epithelioid histology, stage III, and ECOG performance status 3 pre-ITP were prognostic factors with a significant influence on OS. Median OS, calculated from the diagnosis of MPM, was 26.5 months (IQR 22.5-28). ITP is safe, tolerable, and useful but its inclusion in the multidisciplinary palliative treatment of progressive MPM patients should be investigated in a larger multicentre controlled study.


Impaired intracortical inhibition demonstrated in vivo in people with Dravet syndrome.
Dravet syndrome is a rare neurodevelopmental disorder characterized by seizures and other neurologic problems. SCN1A mutations account for ∼80% of cases. Animal studies have implicated mutation-related dysregulated cortical inhibitory networks in its pathophysiology. We investigated such networks in people with the condition. Transcranial magnetic stimulation using single and paired pulse paradigms was applied to people with Dravet syndrome and to 2 control groups to study motor cortex excitability. Short interval intracortical inhibition (SICI), which measures GABAergic inhibitory network behavior, was undetectable in Dravet syndrome, but detectable in all controls. Other paradigms, including those testing excitatory networks, showed no difference between Dravet and control groups. There were marked differences in inhibitory networks, detected using SICI paradigms, while other inhibitory and excitatory paradigms yielded normal results. These human data showing reduced GABAergic inhibition in vivo in people with Dravet syndrome support established animal models.


Quality of life varies with gender and age among adults treated for unilateral cleft lip and palate.
To evaluate health-related quality of life among adults treated for unilateral cleft lip and palate and to investigate whether patients were affected differently depending on gender and age. Cross-sectional population study with long-term follow-up. PATIENTS/SETTINGS: All unilateral cleft lip and palate patients born 1960-1987 and treated at Uppsala University Hospital were invited (n = 109). Response rate was 79% (n = 86). Mean follow-up time was 35 years. Norm data matched for age and gender were used for comparison (n = 1385). Health-related quality of life measured with the SF-36 questionnaire. High values indicate good level of health-related quality of life. The total patient group had lower values in the Mental Health subscale compared with norm data (p = .005). Values in all other subscales did not differ from norm data. Women had a higher positive difference than men in the subscale emotional role function compared with the matched norm population (p < .001). The younger age group (20 to 32 years old) consistently had a larger negative difference to matched norm data compared with the older age group (33 to 47 years old) in the subscales social function (p = .009), physical role function (p < .001), and emotional role function (p < .001). Unilateral cleft lip and palate affected health-related quality of life differently depending on gender and age of the patient. Younger patients were affected more negatively than older patients on several subscales. However, except for the mental health subscale, health-related quality of life was similar among unilateral cleft lip and palate patients and norm data.


Toward a model for assessing level of personality functioning in DSM-5, part II: empirical articulation of a core dimension of personality pathology.
The extensive comorbidity among Diagnostic and Statistical Manual of Mental Disorders (4th ed. [DSM-IV]; American Psychiatric Association, 1994 ) personality disorders might be compelling evidence of essential commonalities among these disorders reflective of a general level of personality functioning that in itself is highly relevant to clinical decision making. This study sought to identify key markers of such a level, thought to reflect a core dimension of personality pathology involving impairments in the capacities of self and interpersonal functioning, and to empirically articulate a continuum of severity of these problems for DSM-5. Using measures of hypothesized core dimensions of personality pathology, a description of a continuum of severity of personality pathology was developed. Potential markers at various levels of severity of personality pathology were identified using item response theory (IRT) in 2 samples of psychiatric patients. IRT-based estimates of participants' standings on a latent dimension of personality pathology were significantly related to the diagnosis of DSM-IV personality disorder, as well as to personality disorder comorbidity. Further analyses indicated that this continuum could be used to capture the distribution of pathology severity across the range of DSM-IV personality disorders. The identification of a continuum of personality pathology consisting of impairments in self and interpersonal functioning provides an empirical foundation for a "levels of personality functioning" rating proposed as part of a DSM-5 personality disorder diagnostic formulation.


Clonal expansion of a virulent Streptococcus suis serotype 9 lineage distinguishable from carriage subpopulations.
Streptococcus suis is a porcine pathogen, causing severe invasive infections. S. suis serotype 9 is increasingly causing disease in Dutch and Chinese pig herds, but it is unknown whether all serotype 9 isolates are equally virulent and markers that can identify virulent strains are not available. Therefore, discrimination between virulent isolates and carriage isolates typically not associated with disease, is currently not possible. We collected tonsillar S. suis isolates from 6 herds not previously diagnosed with S. suis infections, and clinical S. suis isolates of previously diseased pigs. We confirmed the virulence of a virulent type strain and one representative clinical isolate, and the lack of virulence of two carriage isolates, in a pig infection model. Phylogenetic analysis of whole genome sequences of 124 isolates resulted in 10 groups, of which two were almost uniquely populated by clinical isolates. The population structure of S. suis serotype 9 appears highly diverse. However, analysis of the capsule loci sequences showed variation in a single region which fully correlated with a virulent genotype. Transmission electron microscopy suggested differences in capsule thickness between carriage and clinical genotypes. In conclusion, we found that that the S. suis serotype 9 population in the Netherlands is diverse. A distinct virulence-associated lineage was identified and could be discriminated based on the capsule locus sequence. Whilst the difference in virulence cannot be directly attributed to the DNA sequence, the correlation of capsule locus sequence with virulence could be used in the development of diagnostic tests to identify potential virulent S. suis serotype 9 in pigs.


Flavobacterium commune sp. nov., isolated from freshwater and emended description of Flavobacterium seoulense.
A Gram-stain-negative, yellow, facultatively-anaerobic, short, rod-shaped, non-spore-forming bacterium, designated PK15T, was isolated from freshwater. Growth was observed at 4-40 °C (optimum, 30 °C), pH 6-9 (optimum, 8), and in the presence of 0-0.8 % (w/v) NaCl (optimum, 0.4 %). Strain PK15T exhibited both catalase and oxidase activities and was able to reduce nitrate. On the basis of 16S rRNA gene sequence similarities, strain PK15T was shown to belong to the genus Flavobacterium with close similarities to Flavobacterium palustre S44T (97.9 %) and Flavobacterium seoulense EM1321T (97.7 %). Menaquinone-6 (MK-6) was the major respiratory quinone, while the G+C content of the genomic DNA was 35.5 (±0.9) mol%. The major polar lipids were phosphatidylethanolamine, three unidentified aminolipids, one unidentified aminophospholipid and three unidentified polar lipids. The predominant cellular fatty acids (≥10 %) were anteiso-C15 : 0 (17.3 %), a summed feature comprising C16 : 1ω7c and/or C16 : 1ω6c (15.1 %) and iso-C15 : 0 (10.0 %). Chemotaxonomic data supported the affiliation of strain PK15T to the genus Flavobacterium. The results of physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain PK15T from strains of closely related species. It was, therefore, evident that PK15T represents a novel species of the genus Flavobacterium, for which the name Flavobacterium commune sp. nov. is proposed with strain PK15T (=KCTC 52562T=JCM 32115T) as the type strain. Based on the results of the chemotaxonomic characterization in the present study, an emended description of Flavobacterium seoulense is also proposed.


Cytomegalovirus reactivation during alemtuzumab therapy for chronic lymphocytic leukemia: incidence and treatment with oral ganciclovir.
Although alemtuzumab (campath-1H) has been successfully used in patients with untreated or previously treated chronic lymphocytic leukemia (CLL), a variable incidence of cytomegalovirus (CMV) reactivation has been described. No prospective reports currently provide results of the use of oral ganciclovir as pre-emptive therapy in patients with CMV reactivation during alemtuzumab treatment. We designed a prospective study in 12 patients with pretreated CLL with the aim of evaluating the incidence of CMV reactivation during alemtuzumab treatment and the role of oral ganciclovir as pre-emptive therapy and in preventing CMV organ disease. In the 12 CLL patients being treated with alemtuzumab, 8 patients (66%) had CMV reactivation, as detected by antigenemia and/or CMV DNA. No patient showed clinical evidence of CMV disease. The alemtuzumab was discontinued and the patients were immediately treated with oral ganciclovir 1000 mg tid. After a median of 14 days of antiviral therapy all patients achieved negative CMV polymerase chain reaction (PCR) assays and/or antigenemia. No patients showed further CMV reactivation up to the end of the study. CMV reactivaction, studied with periodic analysis of antigenemia and PCR, is frequent in previously treated CLL patients receiving alemtuzumab therapy although only sporadic cases of CMV disease have been reported. Using oral ganciclovir, the response to therapy was prompt, there was no progression to CMV disease, and no relevant clinical toxicity, thus sparing unnecessary hospitalization. Oral ganciclovir may be used as pre-emptive therapy in all patients who develop CMV reactivation during alemtuzumab treatment.


Ceftolozane/tazobactam activity tested against aerobic Gram-negative organisms isolated from intra-abdominal and urinary tract infections in European and United States hospitals (2012).
Ceftolozane/tazobactam is under clinical development for treatment of complicated intra-abdominal infections (IAI), complicated urinary tract infections (UTI) and ventilator-associated pneumonia. We evaluated the in vitro activity of ceftolozane/tazobactam and comparator agents tested against Gram-negative aerobic bacteria causing IAI and healthcare-associated UTI (HCA-UTI). The organisms were consecutively collected from January to December 2012 from 59 medical centers located in the United States (USA) and 15 European countries by the Program to Assess Ceftolozane/Tazobactam Susceptibility (PACTS). The collection included 809 organisms from IAI and 2474 organisms from HCA-UTI, and susceptibility testing was performed by reference broth microdilution methods as described by the Clinical and Laboratory Standards Institute (CLSI) M07-A9 document. Overall, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa were the most frequently isolated pathogens from both infection types. Ceftolozane/tazobactam was very active against E. coli (MIC50/90, 0.25/0.5 mg/L; 98.5-99.9% inhibited at an MIC of ≤8 mg/L) and retained activity against many of the multidrug-resistant (MDR; MIC50/90, 0.5/2->32 mg/L) and ESBL-phenotype strains (MIC50/90, 0.5/2-32 mg/L). Ceftolozane/tazobactam was active against most K. pneumoniae strains (MIC50/90, 0.25/16 mg/L, 88.9-89.6% inhibited at an MIC of ≤8 mg/L), but some ESBL-phenotype (MIC50/90, 4-8/>32 mg/L) and MDR (MIC50/90, 16/>32 mg/L) isolates exhibited elevated MIC values. Ceftolozane/tazobactam was the most active agent tested against P. aeruginosa (MIC50/90, 0.5/4 mg/L; 93.4-95.7% inhibited at ≤8 mg/L) and retained potency against many MDR (MIC50/90, 2-4/>32 mg/L), ceftazidime-nonsusceptible (MIC50/90, 2-4/>32 mg/L) and meropenem-nonsusceptible (MIC50/90, 2/>32 mg/L) strains. Ceftolozane/tazobactam was also active against Klebsiella oxytoca (MIC50/90, ≤0.12-0.25/0.5-1 mg/L), Enterobacter spp. (MIC50/90, 0.25-0.5/4-8 mg/L), Citrobacter spp. (MIC50/90, 0.25/2-32 mg/L), Proteus mirabilis (MIC50/90, 0.5/0.5 mg/L), indole-positive Proteae (MIC50/90, 0.25/0.5-1 mg/L), and Serratia spp. (MIC50/90, 0.5/1-2 mg/L). In summary, ceftolozane/tazobactam demonstrated potent in vitro activity when tested against contemporary aerobic Gram-negative pathogens causing IAI and HCA-UTI in USA and European medical centers.


Establishment and characterization of a new human cultured cell line from a prolactin-secreting pituitary adenoma.
A new human cell line was established from a prolactin (PRL) secreting pituitary adenoma. This cell line, designated as HPA, initially produced and secreted PRL, but the ability was decreased with increasing passage number. After about 30 passages in vitro, these cells had a short doubling time (14 h) and a low plating efficiency (9%). When a minimum of 10(5) cells was injected per mouse, virtually all athymic nude mice developed a slow growing, nonmetastasizing tumor at the injection site about 30 days after injection. PRL production by the HPA cells after Day 150 was demonstrated by immunocytochemistry as well as radioimmunoassay. In addition, cimetidine (10(-4) M) had a significant stimulatory effect on PRL secretion by 4-day-cultured HPA cells. When the HPA cells were incubated in the presence of 5.0 and 10.0 nM bromocriptine, the proliferation rate was inhibited to 53.4 and 25.1% of untreated controls, respectively. On the other hand, the same concentrations of bromocriptine did not affect the proliferation rate of YK cells derived from human immature teratoma of the ovary. In addition, bromocriptine inhibited significantly the growth rate of xenotransplanted HPA but not YK cells. These results suggest that bromocriptine inhibits specifically the proliferation of HPA cells.


Ralfuranone thioether production by the plant pathogen Ralstonia solanacearum.
Ralfuranones are aryl-substituted furanone secondary metabolites of the Gram-negative plant pathogen Ralstonia solanacearum. New sulfur-containing ralfuranone derivatives were identified, including the methyl thioether-containing ralfuranone D. Isotopic labeling in vivo, as well as headspace analyses of volatiles from R. solanacearum liquid cultures, established a mechanism for the transfer of an intact methylthio group from L-methionine or α-keto-γ-methylthiobutyric acid. The methylthio acceptor molecule ralfuranone I, a previously postulated biosynthetic intermediate in ralfuranone biosynthesis, was isolated and characterized by NMR. The highly reactive Michael acceptor system of this intermediate readily reacts with various thiols, including glutathione.


Not just gene expression: 3D implications of chromatin modifications during sexual plant reproduction.
DNA methylation and histone modifications are epigenetic changes on a DNA molecule that alter the three-dimensional (3D) structure locally as well as globally, impacting chromatin looping and packaging on a larger scale. Epigenetic marks thus inform higher-order chromosome organization and placement in the nucleus. Conventional epigenetic marks are joined by chromatin modifiers like cohesins, condensins and membrane-anchoring complexes to support particularly 3D chromosome organization. The most popular consequences of epigenetic modifications are gene expression changes, but chromatin modifications have implications beyond this, particularly in actively dividing cells and during sexual reproduction. In this opinion paper, we will focus on epigenetic mechanisms and chromatin modifications during meiosis as part of plant sexual reproduction where 3D management of chromosomes and re-organization of chromatin are defining features and prime tasks in reproductive cells, not limited to modulating gene expression. Meiotic chromosome organization, pairing and synapsis of homologous chromosomes as well as distribution of meiotic double-strand breaks and resulting crossovers are presumably highly influenced by epigenetic mechanisms. Special mobile small RNAs have been described in anthers, where these so-called phasiRNAs seem to direct DNA methylation in meiotic cells. Intriguingly, many of the mentioned developmental processes make use of epigenetic changes and small RNAs in a manner other than gene expression changes. Widening our approaches and opening our mind to thinking three-dimensionally regarding epigenetics in plant development holds high promise for new discoveries and could give us a boost for further knowledge.


Weber's law for biological responses in autocatalytic networks of chemical reactions.
Biological responses often obey Weber's law, according to which the magnitude of the response depends only on the fold change in the external input. In this study, we demonstrate that a system involving a simple autocatalytic reaction shows such a response when a chemical is slowly synthesized by the reaction from a faster influx process. We also show that an autocatalytic reaction process occurring in series or in parallel can obey Weber's law with an oscillatory adaptive response. Considering the simplicity and ubiquity of the autocatalytic process, our proposed mechanism is thought to be commonly observed in biological reactions.


The introduction of navigation in liver surgery in Brazil.
The authors thoroughly report the development, the technical aspects and the performance of the first navigated liver resections, by laparotomy and laparoscopy, in Brazil, done at the National Cancer Institute, Ministry of Health, using a surgical navigator.


How useful is thyroid function testing in patients with recent-onset atrial fibrillation? The Canadian Registry of Atrial Fibrillation Investigators.
Patients with recent-onset atrial fibrillation often undergo routine thyroid function screening to rule out thyroid disease as a cause of atrial fibrillation. Patients with recent (< 3 months) onset of documented atrial fibrillation or flutter were enrolled in the Canadian Registry of Atrial Fibrillation from outpatient clinics, emergency departments, and hospital wards across Canada. Seven hundred twenty-six patients underwent baseline thyroid function screening and were assessed for presence of clinical thyroid disease. Serum thyrotropin level (TSH) was measured in 707 patients (97%), and thyroxine level (T4) in 407 patients (56%). A TSH level less than 0.1 mU/L was present in 5 patients (0.7%). A TSH level less than normal but more than 0.1 mU/L was present in 34 patients (4.7%). No patient had definite hypothyroidism (TSH > 20 mU/L), but 56 patients (7.7%) had an elevated TSH level that was less than 20 mU/L. During 1.7 years of follow-up, only 7 patients were found to have clinical hyperthyroidism, and 11 patients (1.5%) had hypothyroidism. Logistic regression analysis showed that palpitations (odds ratio, 4.9; 95% confidence interval, 1.7-14.0) and asymptomatic presentation (odds ratio, 5.5; 95% confidence interval, 1.9-16.2) were risk factors for low TSH level, and increasing age (odds ratio, 1.32 every 10 years; 95% confidence interval, 1.01-1.66) was a risk factor for high TSH level. The positive predictive value of palpitations and asymptomatic presentation for low TSH level were 9% and 8%, respectively. An abnormal TSH level is common in patients with recent-onset atrial fibrillation. However, clinical thyroid disease is uncommon. Routine TSH screening of patients who have atrial fibrillation has a low yield and may be better applied to those patients at higher risk of having undiagnosed clinical thyroid disease.


Wild-type opaque2 and defective opaque2 polypeptides form complexes in maize endosperm cells and bind the opaque2-zein target site.
The Opaque2 (O2) basic leucine (Leu)-zipper transcriptional activator controls the expression of several genes in maize (Zea mays). We investigated the phosphorylation extent of wild-type O2 and mutant-defective or mutant-truncated o2 polypeptides in endosperm cells, their subcellular localization, participation in complex formation, and involvement in functional activity. Besides wild type, four mutant alleles (o2T, o2-52, o2It, and o2-676) producing o2 polypeptides and a null transcript allele (o2R) were considered. Observing the effects of these mutations, multiphosphorylation events in O2 or o2 proteins were confirmed and further investigated, and the involvement of both the nuclear localization signal (NLS)-B and Leu-zipper domains in proper targeting to the nucleus was ascertained. The absence of these domains in the o2T and o2It-S mutant-truncated forms holds them within the cytoplasm, where they are partially phosphorylated, whereas the presence of NLS-B and a partial Leu-zipper domain in o2-52 distributes this mutant-truncated form in both cytoplasm and nucleus. Although mutated in the NLS-B domain, the o2It-L and o2-676 mutant-defective forms are, respectively, partially or completely distributed into the nucleus. Only wild-type O2 and mutant-defective o2 polypeptides bearing the Leu-zipper are able to form complexes whose components were proven to bind the O2-zein target site by in vitro analyses. The transcription of a subset of H-zein genes as well as H-zein polypeptide accumulation in several o2-mutant-defective genotypes indicate the in vivo involvement of o2-mutant-defective proteins in O2-zein target site recognition. The gathered information broadens our knowledge on O2 functional activity and our view on possible quality protein maize trait manipulation or plant transformation via the utilization of cisgenic elements.


[Sleeping position for preterm infants].
Supine sleeping is recommended to prevent the sudden infant death syndrome (SIDS). Low birth weight infants are at increased risk for SIDS, which is increased further if they are placed prone. Prone sleeping, however, also has advantages for preterm infants, such as a reduced apnoea rate, an increased lung volume and more quiet sleep. In their first weeks of life, these infants are usually on a monitor and under continuous observation. SIDS is extremely unlikely under these circumstances. Because of the aforementioned advantages, these infants may be placed prone during their first few weeks of life in the hospital. One week before discharge, however, they should be changed to back sleeping and the parents be explained that their baby is now nearing discharge and should thus be placed as it should also sleep at home: on its back and in a sleeping sack. We do not prescribe home monitors for SIDS prevention, but occasionally use pulse oximeters at home for a few weeks if an infant continues to exhibit apnoea of prematurity.


The 9-lipoxygenase Osr9-LOX1 interacts with the 13-lipoxygenase-mediated pathway to regulate resistance to chewing and piercing-sucking herbivores in rice.
Oxylipins produced by the 13-lipoxygenase (LOX) have been reported to play an important role in plant defense responses to herbivores. Yet, the role of oxylipins produced by the 9-LOX pathway in this process remains largely unknown. Here we cloned a gene encoding a chloroplast-localized 9-LOX, Osr9-LOX1, from rice. Transcriptional analysis revealed that herbivore infestation, mechanical wounding and jasmonic acid (JA) treatment either repressed or did not enhance the level of Osr9-LOX1 transcripts at early stages but did at later stages, whereas salicylic acid (SA) treatment quickly increased the transcript level of Osr9-LOX1. Antisense expression of Osr9-lox1 (as-r9lox1) decreased the amount of wound-induced (Z)-3-hexenal but increased levels of striped stem borer (SSB)-induced linolenic acid, JA, SA and trypsin protease inhibitors. These changes were associated with increased resistance in rice to the larvae of the SSB Chilo suppressalis. In contrast, although no significant differences were observed in the duration of the nymph stage or the number of eggs laid by female adults between the brown planthopper (BPH) Nilaparvata lugens that fed on as-r9lox1 lines and BPH that fed on wild-type (WT) rice plants, the survival rate of BPH nymphs that fed on as-r9lox1 lines was higher than that of nymphs that fed on WT plants, possibly because of a higher JA level. The results demonstrate that Osr9-LOX1 plays an important role in regulating an herbivore-induced JA burst and cross-talk between JA and SA, and in controlling resistance in rice to chewing and phloem-feeding herbivores.


Psychopathology and history of mental healthcare among male detainees transferred to a facility for managing otherwise uncontrollable in-prison violence: An exploratory study.
In-prison violence by detainees is a problem worldwide, but despite evidence of a much higher prevalence of a range of psychiatric disorders than in the general population, little is known about psychopathology among violent detainees. Our aim was to explore the psychopathology and mental healthcare history of Dutch detainees who were transferred to the highly restrictive facility for uncontrollably violent detainees following severe in-prison violence. Anonymised data for all 253 male detainees incarcerated at any time between January 2016 and January 2020 in the specialist national facility for those seriously violent while in prison-'the Violence Facility'-were obtained from the Dutch Ministry of Justice together with similarly recorded data for a matched comparison group of 253 detainees admitted to an in-prison psychiatric facility-'the Psychiatric Facility'. There was no record of any psychiatric assessment for 29% of the Violence Facility men. Almost all of the detainees who had been assessed were classified with at least one disorder. Compared to detainees in the Psychiatric Facility, Violence Facility men were more likely to be diagnosed with attention deficit hyperactivity disorder (ADHD), anxiety, behavioural and personality disorders; Psychiatric Facility men were more likely to be diagnosed with psychosis or substance use disorder. Most men in both groups had previously used mental health services. This first study of detainees in the Dutch in-prison facility for violent detainees raises questions about whether the extent of violence among these men may have masked mental healthcare needs and leads to questions about potential benefits from establishing more systematic mental health assessments for them, and a need for more specialist services.


Effects of progesterone on blood pressure, plasma volume, and responses to hypotension.
Gonadal steroids have been implicated in the control of blood pressure and fluid homeostasis. These experiments test the effect of progesterone in ovariectomized ewes on blood pressure, volume, and hormone responses to hypotension. Eight ewes were each studied in four conditions: ovariectomized, progesterone and estrogen replaced, progesterone replaced, or sham treated. During each treatment mean arterial pressure (MAP), plasma volume (PV), baroreflex responsiveness, and adrenocorticotropic hormone (ACTH), arginine vasopressin (AVP), and renin responses to hypotension were determined. Progesterone treatment significantly reduced resting MAP and increased PV compared with ovariectomy or sham treatments. Heart period at 85 mmHg was reduced with progesterone treatment. There was no effect of progesterone treatment on ACTH, AVP, or renin or heart rate responses to hypotension. Basal AVP levels were increased, and angiotensin II concentrations were decreased, by estrogen and progesterone and by sham treatments; plasma Na+ also tended to be increased during these treatments. These results suggest that a small increase in progesterone can reset resting MAP and PV without altering reflex heart rate or endocrine responses to hypotension.


Survey of multidrug resistance integrative mobilizable elements SGI1 and PGI1 in Proteus mirabilis in humans and dogs in France, 2010-13.
To characterize MDR genomic islands related to Salmonella genomic island 1 (SGI1) and Proteus genomic island 1 (PGI1) in Proteus mirabilis from human and animal sources in France in light of the previously reported cases. A total of 52 and 46 P. mirabilis clinical strains from human and animal sources, respectively, were studied for the period 2010-13. MDR was assessed by antimicrobial susceptibility testing, PCR detection of SGI1 and PGI1 and PCR mapping of the MDR regions. The diversity of the SGI1/PGI1-positive P. mirabilis strains was assessed by PFGE. Twelve P. mirabilis strains (5 humans and 7 dogs) were found to harbour an MDR island related to SGI1 or PGI1. Among them, several SGI1 variants were identified in diverse P. mirabilis genetic backgrounds. The variant SGI1-V, which harbours the ESBL bla VEB-6 gene, was found in closely genetically related human and dog P. mirabilis strains. The recently described PGI1 element was also identified in human and dog strains. Finally, one strain harboured a novel SGI genomic island closely related to SGI1 and SGI2 without an insertion of the MDR region. This study reports for the first time, to our knowledge, SGI1-positive and PGI1-positive P. mirabilis strains from dogs in France. The genetic diversity of the strains suggests several independent horizontal acquisitions of these MDR elements. The potential transmission of SGI1/PGI1-positive P. mirabilis strains between animals and humans is of public health concern, notably with regard to the spread of ESBL and carbapenemase genes, i.e. bla VEB-6 and bla NDM-1.


Bulk and surface calorimetric measurements at CO wavelengths.
Laser calorimetry was used to measure the optical absorption of several candidate window materials for application at CO laser wavelengths. These materials include KCl, CaF(2), SrF(2), and ZnSe. The long, thin bar technique was used to separate bulk and surface absorption contributions by means of their time dependencies. One sample of KCl, known to be low absorbing at 10.6 microm, exhibited no measurable absorption within the sensitivity of the calorimeter (beta(t) < 2 x 10(-6) cm(-1)). The absorption coefficients measured for two samples of CaF(2) at 5.41 microm were higher than those measured by other investigators at 5.25 microm, but, when fitted to an exponential dependence on wavelength, compared favorably with currently measured values. To compare results from different laboratories, precise definition of the spectral power distribution is essential. The surface absorption was, in all cases, small relative to similar measurements at 10.6 microm; in several cases it was not separable from bulk absorption by the technique used.


Invasive competitor and native seed predators contribute to rarity of the narrow endemic Astragalus sinuatus Piper.
The conservation of rare plant species hinges on our ability to identify the underlying mechanisms that limit rare plant populations. Theory on rarity suggests that both predispersal seed predation and competition can be important mechanisms influencing abundance and/or distribution of rare plant populations. Yet few studies have tested these interactions, and those that have evaluated each mechanism independently. Astragalus sinuatus Piper (Whited's milkvetch) is a narrow endemic plant species restricted to eight populations within a 10-km2 area in eastern Washington. We used experimental and observational methods to test the effects of native insect predispersal seed predators and an invasive grass (Bromus tectorum L. [cheatgrass]) on seed set and population density of A. sinuatus. We quantified per capita seed production and pod predation rates across four sites and among four years. Seed predation rates were high across four sites (66-82%) and all years (65-82%). Experimental reduction of predispersal seed predators significantly increased per capita seed set of A. sinuatus (164-345%) at two experimental sites. Concurrently, two seed addition experiments demonstrated the effect of seed loss and presence of B. tectorum on seedling recruitment and establishment of A. sinuatus over four growing seasons. In the first seed addition experiment, we found no difference in recruitment and establishment between low (40) and high (120) seed addition levels. In the second addition experiment (one level of addition; 40 seeds), we found that recruitment and survivorship increased 200% in plots where B. tectorum was removed compared to plots where B. tectorum was present. Thus, seed addition had no impact in the presence of B. tectorum; conversely, in the absence of B. tectorum, seed addition was highly effective at increasing population numbers. Results suggest that, in areas where B. tectorum is present, recruitment is site limited, and it is seed limited when B. tectorum is absent. We recommend that managers reduce B. tectorum in an effort to increase population growth of A. sinuatus; in areas where B. tectorum is absent, short-term reduction of insect predators should be considered as a strategy to increase population growth of this rare species.


PET Imaging of Macrophage Mannose Receptor-Expressing Macrophages in Tumor Stroma Using 18F-Radiolabeled Camelid Single-Domain Antibody Fragments.
Tumor-associated macrophages constitute a major component of the stroma of solid tumors, encompassing distinct subpopulations with different characteristics and functions. We aimed to identify M2-oriented tumor-supporting macrophages within the tumor microenvironment as indicators of cancer progression and prognosis, using PET imaging. This can be realized by designing (18)F-labeled camelid single-domain antibody fragments (sdAbs) specifically targeting the macrophage mannose receptor (MMR), which has been identified as an important biomarker on this cell population. Cross-reactive anti-MMR sdAbs were generated after immunization of an alpaca with the extracellular domains of both human and mouse MMR. The lead binder was chosen on the basis of comparisons of binding affinity and in vivo pharmacokinetics. The PET tracer (18)F-fluorobenzoate (FB)-anti-MMR sdAb was developed using the prosthetic group N-succinimidyl-4-(18)F-fluorobenzoate ((18)F-SFB), and its biodistribution, tumor-targeting potential, and specificity in terms of macrophage and MMR targeting were evaluated in mouse tumor models. Four sdAbs were selected after affinity screening, but only 2 were found to be cross-reactive for human and mouse MMR. The lead anti-MMR 3.49 sdAb, bearing an affinity of 12 and 1.8 nM for mouse and human MMR, respectively, was chosen for its favorable in vivo biodistribution profile and tumor-targeting capacity. (18)F-FB-anti-MMR 3.49 sdAb was synthesized with a 5%-10% radiochemical yield using an automated and optimized protocol. In vivo biodistribution analyses showed fast clearance via the kidneys and retention in MMR-expressing organs and tumor. The kidney retention of the fluorinated sdAb was 20-fold lower than a (99m)Tc-labeled counterpart. Compared with MMR- and C-C chemokine receptor 2-deficient mice, significantly higher uptake was observed in tumors grown in wild-type mice, demonstrating the specificity of the (18)F tracer for MMR and macrophages, respectively. Anti-MMR 3.49 was denoted as the lead cross-reactive MMR-targeting sdAb. (18)F radiosynthesis was optimized, providing an optimal probe for PET imaging of the tumor-promoting macrophage subpopulation in the tumor stroma.


Challenges impeding integration of oral health into primary health care.
The primary healthcare (PHC) services in the Islamic Republic of Iran have succeeded in addressing high levels of communicable diseases; however, they seem less able to deal with maternal and paediatric oral diseases. The aim of this study was to examine problems in integrating oral health services into PHC. This was a qualitative research study comprising focus group discussions and interviews. Five focus-group discussions were held with midwives, family healthcare practitioners, rural PHC workers, duty-service dentists, and public health dentists. Also, individual interviews were organized with experts of faculty members in related fields, informant managers and policy makers, and in-depth interviews were done with pregnant women in four PHC centres. Audiotapes were transcribed following each session, and then a qualitative thematic analysis was carried out on gathered data. Data analysis resulted in 4 main themes relating to the challenges: environmental, educational, organizational and school-based programme factors. This study provides a clearer understanding of the challenges of integrating oral health services into PHC.


Finally, fixing health care: what's different now?
Is now the time to fix the U.S. health care system? Those who remember the failed attempts of the past would say no. We see it differently. Our optimism is rooted in new developments that didn't exist the last time Congress addressed health care. These include bipartisan support for our Healthy Americans Act; an ideological truce over the role of government in health care; common ground between business and labor; the realization that states can't go it alone on health care; the plight of employers in a global marketplace; and the need for coverage that is affordable, accessible, and portable.


Longitudinal Changes in Sitting Patterns, Physical Activity, and Health Outcomes in Adolescents.
This study examined two-year changes in patterns of activity and associations with body mass index (BMI) and waist circumference (WC) among adolescents. Inclinometers (activPAL) assessed sitting, sitting bouts, standing, stepping, and breaks from sitting. ActiGraph-accelerometers assessed sedentary time (SED), light-intensity physical activity (LIPA, stratified as low- and high-LIPA), and moderate-to-vigorous physical activity (MVPA). Anthropometric measures were objectively assessed at baseline and self-reported at follow-up. Data from 324 and 67 participants were obtained at baseline and follow-up, respectively. Multilevel mixed-effects linear regression models examined changes over time, and associations between baseline values and BMI and WC at follow-up. There were significant increases in BMI (0.6 kg/m²) and durations of prolonged sitting (26.4 min/day) and SED (52 min/day), and significant decreases in stepping (-19 min/day), LIPA (-33 min/day), low-LIPA (-26 min/day), high-LIPA (-6.3 min/day), MVPA (-19 min/day), and the number of breaks/day (-8). High baseline sitting time was associated (p = 0.086) with higher BMI at follow-up. There were no significant associations between baseline sitting, prolonged sitting, LIPA, or MVPA with WC. Although changes in daily activity patterns were not in a favourable direction, there were no clear associations with BMI or WC. Research with larger sample sizes and more time points is needed.


Is rehabilitation of patients with chronic bronchial asthma possible.
Based on his own experience and on the data in the literature the author considers that rehabilitation of adults with chronic bronchial asthma is possible at any stage of disease if there are no complications or morbid associations to prevent it. Rehabilitation of chronic asthmatic patients requires the close collaboration of several specialists such as allergologists, chest physicians, psychologists, physical therapists, balneologists, etc. Rehabilitation in asthmatic patients may only be considered successful if they resume full-time work (in case they had been in receipt of compensation), if the medical leaves are fewer and shorter and if the drug therapy is reduced or null. The author considers that since bronchial asthma in the whole population of Romania has an incidence of 2.3-2.9% rehabilitation, though costly and sometimes "desperate", is still worth while trying for its medical, economical and psychological implications.


Nrf2 prevents Notch-induced insulin resistance and tumorigenesis in mice.
Insulin resistance is associated with increased incidence and enhanced progression of cancers. However, little is known about strategies that can effectively ameliorate insulin resistance and consequently halt cancer progression. Herein, we propose that the transcription factor Nrf2 (also known as Nfe2l2) may be such a target, given its central role in disease prevention. To this end, we developed a mouse that overexpresses the Notch intracellular domain in adipocytes (AdNICD), leading to lipodystrophy-induced severe insulin resistance and subsequent development of sarcomas, as a model reflecting that Notch signaling is deregulated in cancers and shows positive associations with insulin resistance and fatty liver disease in humans. Nrf2 pathway activation was achieved by knocking down Keap1, a repressor of Nrf2, in the AdNICD background. Constitutively enhanced Nrf2 signaling in this setting led to prevention of hepatic steatosis, dyslipidemia, and insulin resistance by repressing hepatic lipogenic pathways and restoration of the hepatic fatty acid profile to control levels. This protective effect of Nrf2 against diabetes extended to significant reduction and delay in sarcoma incidence and latency. Our study highlights that the Nrf2 pathway, which has been induced by small molecules in clinical trials, is a potential therapeutic target against insulin resistance and subsequent risk of cancer.


miR-195 Inhibits Tumor Progression by Targeting RPS6KB1 in Human Prostate Cancer.
To investigate the involvement of hsa-miRNA-195-5p (miR-195) in progression and prognosis of human prostate cancer. qRT-PCR was performed to detect miR-195 expression in both prostate cancer cell lines and clinical tissue samples. Its clinical significance was statistically analyzed. The roles of miR-195 and its candidate target gene, ribosomal protein S6 kinase, 70 kDa, polypeptide 1 (RPS6KB1) in prostate cancer progression were confirmed on the basis of both in vitro and in vivo systems. miR-195 downregulation in prostate cancer tissues was significantly associated with high Gleason score (P = 0.001), positive metastasis failure (P < 0.001), and biochemical recurrence (BCR, P < 0.001). Survival analysis identified miR-195 as an independent prognostic factor for BCR-free survival of prostate cancer patients (P = 0.022). Then, we confirmed the tumor suppressive role of miR-195 through prostate cancer cell invasion, migration, and apoptosis assays in vitro, along with tumor xenograft growth, angiogenesis, and invasion in vivo according to both gain-of-function and loss-of-function experiments. In addition, RPS6KB1 was identified as a novel direct target of miR-195 through proteomic expression profiling combined with bioinformatic target prediction and luciferase reporter assay. Moreover, the reexpression and knockdown of RPS6KB1 could respectively rescue and imitate the effects induced by miR-195. Importantly, RPS6KB1 expression was closely correlated with aggressive progression and poor prognosis in prostate cancer patients as opposed to miR-195. Furthermore, we identified MMP-9, VEGF, BAD, and E-cadherin as the downstream effectors of miR-195-RPS6KB1 axis. The newly identified miR-195-RPS6KB1 axis partially illustrates the molecular mechanism of prostate cancer progression and represents a novel potential therapeutic target for prostate cancer treatment.


Trust transitivity in social networks.
Non-centralized recommendation-based decision making is a central feature of several social and technological processes, such as market dynamics, peer-to-peer file-sharing and the web of trust of digital certification. We investigate the properties of trust propagation on networks, based on a simple metric of trust transitivity. We investigate analytically the percolation properties of trust transitivity in random networks with arbitrary in/out-degree distributions, and compare with numerical realizations. We find that the existence of a non-zero fraction of absolute trust (i.e. entirely confident trust) is a requirement for the viability of global trust propagation in large systems: The average pair-wise trust is marked by a discontinuous transition at a specific fraction of absolute trust, below which it vanishes. Furthermore, we perform an extensive analysis of the Pretty Good Privacy (PGP) web of trust, in view of the concepts introduced. We compare different scenarios of trust distribution: community- and authority-centered. We find that these scenarios lead to sharply different patterns of trust propagation, due to the segregation of authority hubs and densely-connected communities. While the authority-centered scenario is more efficient, and leads to higher average trust values, it favours weakly-connected "fringe" nodes, which are directly trusted by authorities. The community-centered scheme, on the other hand, favours nodes with intermediate in/out-degrees, in detriment of the authorities and its "fringe" peers.


[Amblyomma tigrinum in the Municipality of Lages, SC, and biology observations under laboratory conditions].
To study the biological features of Amblyomma tigrinum, samples of engorged females collected from dogs in the city of Lages, SC, Brazil, were maintained in Petri dishes into an incubator at 27+/-1 degrees C and 80+/-10% RH, for egg laying. For the development of the subsequent experimental phases, artificial infestations were performed in rabbits using the technique of the cotton bag glued to the ear's base. The mean period of hatch, feeding and larval molt were 11.67, 5.93 and 10.96 days, respectively, whereas the mean period of feeding and nymph's molt were 12.40 and 16.27 days, respectively. Only one engorged female was obtained, with detachment observed on the 22(nd) day. The longevity periods for larvae, nymphs and adults were 75, 96 and 150 days respectively.


Involving parents from the start: formative evaluation for a large randomised controlled trial with Botswana Junior Secondary School students.
While HIV prevention research conducted among adolescent populations may encounter parental resistance, the active engagement of parents from inception to trial completion may alleviate opposition. In preparation for implementing a large randomised controlled trial (RCT) examining the efficacy of a behavioural intervention targeting adolescent sexual risk behaviours, a formative evaluation was undertaken to assess parental reactions to the proposed trial. Six focus groups were conducted with parents of adolescents (aged 13-17) from rural, peri-urban and urban junior secondary schools in Botswana. Focus groups explored comprehension and acceptability among parents of the forthcoming trial including HSV-2 testing, the return of results to the adolescent (not the parent), trial information materials and the parental consent process. Parents welcomed the study and understood and accepted its moral and ethical considerations. Their reactions regarding return of HSV-2 results only to adolescents (not the parent) were mixed. Parents understood the consent process and most agreed to consent, while indicating their desire to remain informed and involved throughout the RCT. The focus group discussions (FGDs) provided valuable information and insights that helped strengthen the study. As a result of parents' feedback, counselling procedures were strengthened and direct linkages to local services and care were made. Informational materials were revised to increase clarity, and materials and procedures were developed to encourage and support parental involvement and parent-child dialogue. Ultimately, parental feedback led to a decision by the Government of Botswana to allow parents to access their child's HSV-2 test results.


[Risk factors in cesarean section].
Improvement of surgical techniques, innovation, and technological development have increased the frequency of cesarean section. To identify the sociodemographics, obstetrical, attention and medical practice factors of risk most frequently associated to cesarean section. From January to June 2007, we carried out a study of cases and controls with 222 cases of cesarean section and 358 controls of vaginal childbirth in Hospital General no. 6 of Ciudad Juárez. We compared frequency of sociodemographics, obstetrical, attention, and medical practice variables, by means of chi2 and Fisher exact tests; association among these variables and cesarean section was considered with odds ratio. In all cases chosen confidence interval was 95%. Risk factors associated with cesarean section were: maternal age over 28 years, previous cesarean section, complicated pregnancy, fetal suffering, cephalopelvic disproportion, deficient prenatal care; fetal podalic version, oxytocin administration, abnormal amniotic fluid, double- or triple-circle umbilical cord; patient attended by a gynecologist with more than 16 years of experience and by a resident; and medical care in evening shift. There was no association with age, menarche, beginning of sexual relationships, body mass index, smoking, or addictions. Risk factors associated with cesarean section were: previous cesarean section and patient attended by a gynecologist with more than 16 years of experience and by a resident.


The Impact of the SARS-CoV-2 Virus (COVID-19) Pandemic and the Rapid Adoption of Telehealth for Cardiac Rehabilitation and Secondary Prevention Programs in Rural and Remote Australia: A Multi-Method Study.
Centre-based cardiac rehabilitation (CR) programs were disrupted and urged to adopt telehealth modes of delivery during the COVID-19 public health emergency. Previously established telehealth services may have faced increased demand. This study aimed to investigate a) the impact of the COVID-19 pandemic on CR attendance/completion, b) clinical outcomes of patients with cardiovascular (CV) diseases referred to CR and, c) how regional and rural centre-based services converted to a telehealth delivery during this time. A cohort of patients living in regional and rural Australia, referred to an established telehealth-based or centre-based CR services during COVID-19 first wave, were prospectively followed-up, for ≥90 days (February to June 2020). Cardiac rehabilitation attendance/completion and a composite of CV re-admissions and deaths were compared to a historical control group referred in the same period in 2019. The impact of mode of delivery (established telehealth service versus centre-based CR) was analysed through a competitive risk model. The adaption of centre-based CR services to telehealth was assessed via a cross-sectional survey. 1,954 patients (1,032 referred during COVID-19 and 922 pre-COVID-19) were followed-up for 161 (interquartile range 123-202) days. Mean age was 68 (standard deviation 13) years and 68% were male. Referrals to the established telehealth program did not differ during (24%) and pre-COVID-19 (23%). Although all 10 centre-based services surveyed adopted telehealth, attendance (46.6% vs 59.9%; p<0.001) and completion (42.4% vs 75.4%; p<0.001) was significantly lower during COVID-19. Referral during vs pre-COVID-19 (sub hazard ratio [SHR] 0.77; 95% CI 0.68-0.87), and to a centre-based program compared to the established telehealth service (SHR 0.66; 95% CI 0.58-0.76) decreased the likelihood of CR uptake. An established telehealth service and rapid adoption of telehealth by centre-based programs enabled access to CR in regional and rural Australia during COVID-19. However, further development of the newly implemented telehealth models is needed to promote CR attendance and completion.


Determinants of sedentary behavior, motivation, barriers and strategies to reduce sitting time in older women: a qualitative investigation.
Sedentary behavior defined as time spent non-exercising seated or reclining posture has been identified has a health risk and associated with frailty and disablement for older adults. Older adults are the most sedentary segment of society. To date no study has investigated the determinants of sedentary behavior in older adults. This study reports a qualitative investigation of the determinants of sedentary behavior, strategies and motivator to reduce sitting time by structured interviews in a group of community dwelling older women (N = 11, age 65 and over). Older women expressed the view that their sedentary behavior is mostly determined by pain which acts both as an incentive to sit and a motivator to stand up, lack of energy in the afternoon, pressure from direct social circle to sit and rest, societal and environmental typecasting that older adult are meant to sit, lack of environmental facilities to allow activity pacing. This qualitative investigation highlighted some factors that older adults consider determinants of their sedentary behavior. Some are identical to those affecting physical activity (self-efficacy, functional limitations, ageist stereotyping) but some appear specific to sedentary behavior (locus of control, pain) and should be further investigated and considered during intervention design. Tailored interventions that pay attention to the pattern of sedentary behavior of individuals appear to be supported by the views of older women on their sedentary behavior.


Disruption of myofibrillar energy use: dual mechanisms that may contribute to postischemic dysfunction in stunned myocardium.
The abnormalities in regional function produced by myocardial ischemia persist after the ischemic episode resolves. Since a close functional coupling exists between myofibrillar creatine kinase and myosin ATPase, a disruption of this coupling could adversely influence myocardial function and might provide a mechanism for the myocardial dysfunction observed. The purpose of the present study was to determine if an alteration in the activity of creatine kinase associated with the myofibril occurs in the postischemic period. Anesthetized open-chest dogs (n = 6) underwent coronary occlusion for 15 minutes, followed by reperfusion for 15 minutes. In reperfused myocardium, adenine nucleotide content was decreased (72 +/- 10% of nonischemic myocardium, p less than 0.05), documenting the presence of previous ischemia. The creatine phosphate content of reperfused myocardium returned to normal, indicating resumption of myocardial energy production. The creatine kinase activity of purified myofibrils isolated from reperfused myocardium was decreased by 17 +/- 7% compared to that of nonischemic myofibrils (p less than 0.03). In addition, the free adenosine diphosphate concentration in reperfused myocardium was calculated to be 96 microM and was less than the Km of adenosine diphosphate determined for myofibrillar creatine kinase (105 microM). The results suggest two putative mechanisms for disruption of energy use in postischemic myocardium: decreased creatine kinase activity associated with the myofibril, and limitation of substrate necessary for maximal creatine kinase activity.


Morphologic and molecular description of Metopus fuscus Kahl from North America and new rDNA sequences from seven metopids (Armophorea, Metopidae).
Most species in the large ciliate genus Metopus Claparède & Lachmann, 1858 lack detailed descriptions based on modern morphologic and molecular methods. This lack of data for the vast majority of species hampers application of a morphospecies approach to the taxonomy of Metopus and other armophorids. In this report we redescribe the large species, Metopus fuscusKahl, 1927 based on in vivo observation, silver impregnation, scanning electron microscopy, and single-cell 18S rDNA sequencing of a freshwater North American (Idaho) population. Metopus fuscus invariably has a perinuclear envelope of endosymbiotic bacteria not found in other species. Unlike the original description of a single row of coarse granules between ciliary rows, the Idaho population has five loose rows of small interkinetal granules. We discuss the possible importance of this character in metopids. We also provide a phylogenetic analysis including seven other new metopid 18S rDNA sequences: Brachonella spiralis, B. galeata, Metopus laminarius, M. setosus, M. striatus, M. violaceus, Palmarella lata. Metopus fuscus and M. setosus form a fully supported clade, challenging previous morphospecies groupings. We discuss some ambiguities of armophorid morphologic terminology in the earlier literature. Our phylogenetic analysis of Idaho metopids indicates that the genera Metopus and Brachonella are both nonmonophyletic.


Ultrasensitive reporter protein detection in genetically engineered bacteria.
We demonstrate the use of laser-induced fluorescence confocal spectroscopy to measure analyte-stimulated enhanced green fluorescent protein (egfp) synthesis by genetically modified Escherichia coli bioreporter cells. Induction is measured in cell lysates and, since the spectroscopic focal volume is approximately the size of one bioreporter cell, also in individual live bacteria. This is, to our knowledge, the first ever proof-of-concept work utilizing instrumentation with single-molecule detection capability to monitor bioreporter response. Although we use arsenic inducible bioreporters here, the method is extensible to gfp/egfp bioreporters that are responsive to other substances.


Characteristics and Outcomes of Eligible Nonenrolled Patients in a Mechanical Ventilation Trial of Acute Respiratory Distress Syndrome.
Patients eligible for randomized controlled trials may not be enrolled for various reasons. Nonenrollment may affect study generalizability and lengthen the time required for trial completion. To describe characteristics and outcomes of eligible nonenrolled (ENE) patients in a multicenter trial of mechanical ventilation strategies. Within the OSCILLATE trial of high-frequency oscillation (HFO) versus conventional ventilation (CV) in adults with adult respiratory distress syndrome, and with approval from research ethics boards, we collected a minimal dataset on patients who satisfied eligibility criteria but were not enrolled. We categorized ENE patients as ENE-HFO and ENE-CV based on receipt of HFO at any time. We used multivariable logistic regression to assess the association between ENE status and mortality. A total of 548 patients were randomized, and 546 were ENE. The most common reasons for ENE were no consent (42%), physician refusal (24%), missed randomization window (15%), and current HFO use (14%). Compared with randomized patients in respective arms of the trial, ENE-HFO patients were younger and had worse lung injury, whereas ENE-CV patients had lower illness severity. ENE status was independently associated with mortality (adjusted odds ratio, 1.39; 95% confidence interval, 1.06-1.84; P = 0.02), with no significant interaction with ventilation treatment group. Nonenrollment was common, with approximately one ENE patient for every randomized patient. Our study suggests that enrollment in trials of mechanical ventilation may be associated with improved outcomes compared with standard care and highlights the need for prospective tracking and transparent reporting of ENE patients as part of trial management.


Pathophysiology of systemic sclerosis: current understanding and new insights.
Introduction: Systemic sclerosis (SSc) is a complex autoimmune connective tissue disease characterized by chronic and progressive tissue and organ fibrosis with broad patient-to-patient variability. Some risk factors are known and include combination of persistent Raynaud's phenomenon, steroid hormone imbalance, selected chemicals, thermal, or other injuries. Endogenous and/or exogenous environmental trigger/risk factors promote epigenetic mechanisms in genetically primed subjects. Disease pathogenesis presents early microvascular changes with endothelial cell dysfunction, followed by the activation of mechanisms promoting their transition into myofibroblasts. A complex autoimmune response, involving innate and adaptive immunity with specific/functional autoantibody production, characterizes the disease. Progressive fibrosis and ischemia involve skin and visceral organs resulting in their irreversible damage/failure. Progenitor circulating cells (monocytes, fibrocytes), together with growth factors and cytokines participate in disease diffusion and evolution. Epigenetic, vascular and immunologic mechanisms implicated in systemic fibrosis, represent major targets for incoming disease modifying therapeutic approaches. Areas covered: This review discusses current understanding and new insights of SSc pathogenesis, through an overview of the most relevant advancements to present aspects and mechanisms involved in disease pathogenesis. Expert opinion: Considering SSc intricacy/heterogeneity, early combination therapy with vasodilators, immunosuppressive and antifibrotic drugs should successfully downregulate the disease progression, especially if started from the beginning.


Caenorhabditis elegans PRMT-7 and PRMT-9 Are Evolutionarily Conserved Protein Arginine Methyltransferases with Distinct Substrate Specificities.
Caenorhabditis elegans protein arginine methyltransferases PRMT-7 and PRMT-9 are two evolutionarily conserved enzymes, with distinct orthologs in plants, invertebrates, and vertebrates. Biochemical characterization of these two enzymes reveals that they share much in common with their mammalian orthologs. C. elegans PRMT-7 produces only monomethylarginine (MMA) and preferentially methylates R-X-R motifs in a broad collection of substrates, including human histone peptides and RG-rich peptides. In addition, the activity of the PRMT-7 enzyme is dependent on temperature, the presence of metal ions, and the reducing agent dithiothreitol. C. elegans PRMT-7 has a substrate specificity and a substrate preference different from those of mammalian PRMT7, and the available X-ray crystal structures of the PRMT7 orthologs show differences in active site architecture. C. elegans PRMT-9, on the other hand, produces symmetric dimethylarginine and MMA on SFTB-2, the conserved C. elegans ortholog of human RNA splicing factor SF3B2, indicating a possible role in the regulation of nematode splicing. In contrast to PRMT-7, C. elegans PRMT-9 appears to be biochemically indistinguishable from its human ortholog.


Tobacco use in popular movies during the past decade.
The top 50 commercially successful films released per year from 1991 to 2000 were content coded to assess trends in tobacco use over time and attributes of films predictive of higher smoking rates. This observational study used media content analysis methods to generate data about tobacco use depictions in films studied (n = 497). Films are the basic unit of analysis. Once films were coded and preliminary analysis completed, outcome data were transformed to approximate multivariate normality before being analysed with general linear models and longitudinal mixed method regression methods. Tobacco use per minute of film was the main outcome measure used. Predictor variables include attributes of films and actors. Tobacco use was defined as any cigarette, cigar, and chewing tobacco use as well as the display of smoke and cigarette paraphernalia such as ashtrays, brand names, or logos within frames of films reviewed. Smoking rates in the top films fluctuated yearly over the decade with an overall modest downward trend (p < 0.005), with the exception of R rated films where rates went up. The decrease in smoking rates found in films in the past decade is modest given extensive efforts to educate the entertainment industry on this issue over the past decade. Monitoring, education, advocacy, and policy change to bring tobacco depiction rates down further should continue.


PROMIS(®) pediatric self-report scales distinguish subgroups of children within and across six common pediatric chronic health conditions.
To conduct a comparative analysis of eight pediatric self-report scales for ages 8-17 years from the National Institutes of Health (NIH) Patient Reported Outcomes Measurement Information System (PROMIS(®)) in six pediatric chronic health conditions, using indicators of disease severity. Pediatric patients (N = 1454) with asthma, cancer, chronic kidney disease, obesity, rheumatic disease, and sickle cell disease completed items from the PROMIS pediatric mobility, upper extremity functioning, depressive symptoms, anxiety, anger, peer relationships, pain interference, and fatigue self-report scales. Comparisons within the six pediatric chronic health conditions were conducted by examining differences in groups based on the disease severity using markers of severity that were specific to characteristics of each disease. A comparison was also made across diseases between children who had been recently hospitalized and those who had not. In general, there were differences in self-reported health outcomes within each chronic health condition, with patients who had higher disease severity showing worse outcomes. Across health conditions, when children with recent hospitalizations were compared with those who had not been hospitalized in the past 6 months, we found significant differences in the expected directions for all PROMIS domains, except anger. PROMIS measures discriminate between different clinically meaningful subgroups within several chronic illnesses. Further research is needed to determine the responsiveness of the PROMIS pediatric scales to change over time.


Protective effects of Rheum tanguticum polysaccharide against hydrogen peroxide-induced intestinal epithelial cell injury.
To describe the effect of Rheum tanguticum polysaccharide (RTP) on hydrogen peroxide-induced human intestinal epithelial cell injury. Hydrogen peroxide (100 micromol/L) was introduced to induce human intestinal epithelial cell injury. Cells were pretreated with RTP (30,100,300 microg/mL) for 24 h before exposure to hydrogen peroxide. Cell viability was detected by MTT assay and morphological observation. Acridine orange staining and flow cytometry were performed to assess cell apoptosis. Lactate dehydrogenase (LDH) activity, production of malondialdehyde (MDA) and superoxide dismutase (SOD) activity were measured by spectrophotometry with corresponding assay kits. Following exposure to H(2)O(2), a marked decrease in cell survival and SOD activity, increased production of MDA, LDH leakage and cell apoptosis were found. Pretreatment of the cells with RTP could significantly elevate cell survival, SOD activity and decrease the level of MDA, LDH activity and cell apoptosis. RTP may have cytoprotective and anti-oxidant effects against H(2)O(2)-induced intestinal epithelial cell injury by inhibiting cell apoptosis and necrosis. This might be one of the possible mechanisms of RTP for the treatment of ulcerative colitis in rats.


Experimental benznidazole treatment of Trypanosoma cruzi II strains isolated from children of the Jequitinhonha Valley, Minas Gerais, Brazil, with Chagas disease.
Trypanosoma cruzi strains from distinct geographic areas show differences in drug resistance and association between parasites genetic and treatment response has been observed. Considering that benznidazole (BZ) can reduce the parasite burden and tissues damage, even in not cured animals and individuals, the goal is to assess the drug response to BZ of T. cruzi II strains isolated from children of the Jequitinhonha Valley, state of Minas Gerais, Brazil, before treatment. Mice infected and treated with BZ in both phases of infection were compared with the untreated and evaluated by fresh blood examination, haemoculture, polymerase chain reaction, conventional (ELISA) and non-conventional (FC-ALTA) serologies. In mice treated in the acute phase, a significant decrease in parasitaemia was observed for all strains. Positive parasitological and/or serological tests in animals treated during the acute and chronic (95.1-100%) phases showed that most of the strains were BZ resistant. However, beneficial effect was demonstrated because significant reduction (p < 0.05%) and/or suppression of parasitaemia was observed in mice infected with all strains (acute phase), associated to reduction/elimination of inflammation and fibrosis for two/eight strains. BZ offered some benefit, even in not cured animals, what suggest that BZ use may be recommended at least for recent chronic infection of the studied region.


Effectiveness of the implementation of a simple radiation reduction protocol in the catheterization laboratory.
A reduction in radiation doses at the catheterization laboratory, maintaining the quality of procedures is essential. Our objective was to analyze the results of a simple radiation reduction protocol at a high-volume interventional cardiology unit. We analyzed 1160 consecutive procedures: 580 performed before the implementation of the protocol and 580 after it. The protocol consisted in: the reduction of the number of ventriculographies and aortographies, the optimization of the collimation and the geometry of the X ray tube-patient-receptor, the use of low dose-rate fluoroscopy and the reduction of the number of cine sequences using the software "last fluoroscopy hold". There were no significant differences in clinical baseline features or in the procedural characteristics with the exception of a higher percentage of radial approach (30.7% vs 69.6%; p<0.001) and of percutaneous coronary interventions of chronic total occlusions after the implementation of the protocol (2.1% vs 6.7%; p=0,001). Angiographic success was similar during both periods (98.3% vs 99.2%; p=0.2). There were no significant differences between both periods regarding the overall duration of the procedures (26.9 vs 29.6min; p=0.14), or the fluoroscopy time (13.3 vs 13.2min; p=0.8). We observed a reduction in the percentage of procedures with ventriculography (80.9% vs 7.1%; p<0.0001) or aortography (15.4% vs 4.4%; p<0.0001), the cine runs (21.8 vs 6.9; p<0.0001) and the dose-area product (165 vs 71 Gyxcm(2); p<0.0001). With the implementation of a simple radiation reduction protocol, a 57% reduction of dose-area product was observed without a reduction in the quality or the complexity of procedures.