A LYSA Phase Ib Study of Tazemetostat (EPZ-6438) plus R-CHOP in Patients with Newly Diagnosed Diffuse Large B-Cell Lymphoma (DLBCL) with Poor Prognosis Features.
The histone-methyl transferase EZH2, catalytic subunit of the PRC2 complex involved in transcriptional regulation, is mutated in approximately 25% of germinal center B-cell lymphomas. Aberrant proliferative dependency on EZH2 activity can be targeted by the orally available EZH2 inhibitor tazemetostat (EPZ-6438). We report the results of the phase Ib tazemetostat plus R-CHOP combination (NCT02889523), in patients 60 to 80 years of age with newly diagnosed diffuse large B-cell lymphoma. The primary objective of this dose-escalation study was to evaluate the safety of the combination and to determine the recommended phase II dose (RP2D) of tazemetostat. A total of 17 patients were enrolled. During C1 and C2, two dose-limiting toxicities were observed: one grade 3 constipation at 400 mg and one grade 5 pulmonary infection at 800 mg. Grade 3 or more toxicities observed in more than 10% of the patients were constipation (24%), nausea (12%), and hypokalemia (12%). Grade 3 to 4 hematologic adverse events were recorded in 8 patients (47%): neutropenia (47%), leukopenia (29%), anemia (18%), and thrombocytopenia (12%). The tazemetostat RP2D was 800 mg. No organ-oriented toxicity increased with tazemetostat dosage escalation (severity and incidence). At 800 mg, AUC and Cmax of tazemetostat were similar compared with the single-agent study (E7438-G000-101). The RP2D of tazemetostat combined with R-CHOP is 800 mg twice a day. The association presents safety and PK comparable with R-CHOP alone. Preliminary efficacy data are encouraging and further investigations in phase II trial are warranted.


Phased Genotyping-by-Sequencing Enhances Analysis of Genetic Diversity and Reveals Divergent Copy Number Variants in Maize.
High-throughput sequencing (HTS) of reduced representation genomic libraries has ushered in an era of genotyping-by-sequencing (GBS), where genome-wide genotype data can be obtained for nearly any species. However, there remains a need for imputation-free GBS methods for genotyping large samples taken from heterogeneous populations of heterozygous individuals. This requires that a number of issues encountered with GBS be considered, including the sequencing of nonoverlapping sets of loci across multiple GBS libraries, a common missing data problem that results in low call rates for markers per individual, and a tendency for applicability only in inbred line samples with sufficient linkage disequilibrium for accurate imputation. We addressed these issues while developing and validating a new, comprehensive platform for GBS. This study supports the notion that GBS can be tailored to particular aims, and using Zea mays our results indicate that large samples of unknown pedigree can be genotyped to obtain complete and accurate GBS data. Optimizing size selection to sequence a high proportion of shared loci among individuals in different libraries and using simple in silico filters, a GBS procedure was established that produces high call rates per marker (>85%) with accuracy exceeding 99.4%. Furthermore, by capitalizing on the sequence-read structure of GBS data (stacks of reads), a new tool for resolving local haplotypes and scoring phased genotypes was developed, a feature that is not available in many GBS pipelines. Using local haplotypes reduces the marker dimensionality of the genotype matrix while increasing the informativeness of the data. Phased GBS in maize also revealed the existence of reproducibly inaccurate (apparent accuracy) genotypes that were due to divergent copy number variants (CNVs) unobservable in the underlying single nucleotide polymorphism (SNP) data.


MYCOBACTOCIDIN, A NEW ANTIBIOTIC ACTIVE AGAINST MYCOBACTERIA.
Fregnan, G. B. (University of Wisconsin, Madison) and D. W. Smith. Mycobactocidin, a new antiboitic active against mycobacteria. J. Bacteriol. 83:1069-1076. 1962.-A new antibiotic, produced by a strain of Staphylococcus epidermidis, is described. The name mycobactocidin is proposed because of the specificity of the antibiotic against mycobacteria. Methods for extraction and purification of the antibiotic are described. The water-soluble fraction is shown to be active in vitro. No toxicity could be detected in mice after intraperitoneal or subcutaneous injection. Enzymatic hydrolysis suggests a glyco-protein nature for the antibiotic. Mycobactocidin is soluble in distilled water at neutrality but is generally insoluble in organic solvents. The antibiotic is precipitated from aqueous solution by heavy metals. It is stable in the pH range of 2 to 8, but is precipitated at pH 3 to 4. An aqueous solution of the antibiotic is stable for several days at 5 C, and for longer periods if lyophilized and stored at -20 C. It does not pass through a dialysis membrane and its activity is retained after 15 to 20 hr of dialysis at 5 C. The activity was shown to be unrelated to the formation of hydrogen peroxide.


Peripheral versus central mechanisms of the cannabinoid type 2 receptor agonist AM1710 in a mouse model of neuropathic pain.
The CB2 R agonist AM1710, examined in animal models of peripheral neuropathy, is effective in controlling aberrant light touch sensitivity, referred to as mechanical allodynia. However, nonspecific binding of AM1710 to CB1 R, either peripherally or centrally, could be partially responsible for the analgesic effects of AM1710. Thus, we sought to determine in mice whether spinal (intrathecal; i.t.) or peripheral AM1710 administration could lead to anti-allodynia by reducing the protein expression of spinal and dorsal root ganglia (DRG) proinflammatory cytokines and elevating the anti-inflammatory cytokine interleukin-10 (IL-10) in the absence of CB1 R. Macrophage cell cultures were examined to characterize AM1710-mediated suppression of the proinflammatory cytokine tumor necrosis factor-alpha (TNF-α). Either i.p. or i.t. AM1710 reversed CCI-induced mechanical allodynia to sham levels in CB1 R (-/-), (+/-), (+/+) mice. CCI-induced neuropathy decreased IL-10 immunoreactivity (IR) in the dorsal root ganglia (DRG) and the dorsal horn of the spinal cord, with i.t. AM1710 restoring basal IL-10 IR. CCI-induced elevations in proinflammatory cytokine IR were decreased within the spinal cord only after i.t. AM1710 in all mouse genotypes. Meanwhile, within DRG tissue from neuropathic mice, proinflammatory cytokines were decreased following either i.p. or i.t. AM1710. Analysis of cultured supernatants revealed AM1710 decreased TNF-alpha protein. We conclude that CB1 R is dispensable for either peripheral or central anti-allodynic actions of AM1710 in neuropathic mice. Cannabinoid CB2 R agonists produce heightened spinal IL-10 which may be clinically relevant to successfully treat neuropathic pain.


Delta tocotrienol in recurrent ovarian cancer. A phase II trial.
Delta tocotrienol has anti-neoplastic activity as demonstrated in several in-vitro and in-vivo investigations. The effect relies on inhibition of different pathways. It also has antiangiogenic activity, and an additive effect to bevacizumab may be expected. The present study was a phase II trial of bevacizumab combined with tocotrienol in chemotherapy refractory ovarian cancer. The study also included analysis of circulating tumor specific HOXA9 methylated DNA (HOXA9 meth-ctDNA) during treatment. The study included 23 patients. The rate of disease stabilization was 70% with very low toxicity. The median PFS was 6.9 months and the median OS 10.9 months, which is rather high compared to the current literature. A division of the patients according to level of HOXA9 meth-ctDNA already after the first cycle of chemotherapy resulted in two groups of patients with different prognoses. Patients with an increasing level of HOXA9 meth-ctDNA had a median PFS and OS of 1.4 and 4.3 months, respectively, compared to 7.8 and 12 months in the group with stable or decreasing levels. The combination of bevacizumab and tocotrienol is potent in chemotherapy refractory ovarian cancer. The level of HOXA9 meth-ctDNA after one cycle of chemotherapy holds important prognostic information.


Direct carbonylation at a C-H bond in the benzene ring of 2-phenyloxazolines catalyzed by Ru(3)(CO)(12). Scope, limitations, and mechanistic aspects
The ruthenium-catalyzed carbonylation at a C-H bond in the benzene ring of a 2-phenyloxazoline is described. The reaction of 2-phenyloxazolines with CO and ethylene in toluene in the presence of a catalytic amount of Ru(3)(CO)(12) resulted in propionylation at an ortho C-H bond in the benzene ring. The presence of the oxazoline ring on the benzene ring is essential for the carbonylation to proceed. Other heterocycles, such as oxazine, oxazole, and thiazoline rings, also served as acceptable directing groups as did the oxazoline ring. A wide functional group compatibility was observed. The site selectivity of the carbonylation was examined using meta-substituted phenyloxazolines. It was found that the carbonylation took place exclusively at the less-hindered C-H bond, irrespective of the nature of substituents, indicating that the site selectivity was determined by steric factors. The reaction was also applicable, not only to a benzene ring, but also to naphthyl and thiophenyl rings. Olefins such as propene and trimethylvinylsilane in place of ethylene could also be used in the carbonylation reaction, while other olefins, such as 1-hexene, tert-butylethylene, vinylcyclohexane, isoprene, 1,5-hexadiene, cyclohexene, 1, 5-cyclooctadiene, styrene, methyl acrylate, vinyl acetate, allyltrimethylsilane, and triethoxyvinylsilane did not afford the coupling products. An equilibrium between 2-phenyloxazolines, carbon monoxide, and olefins exists on one hand and the corresponding ketones on the other hand, and product composition is governed by the equilibrium thermodynamics of the system. The results of deuterium labeling experiments suggest that the catalysis involves a reversible C-H bond cleavage and that the rate-determining step is not the cleavage of a C-H bond. The results of kinetic study of the effects of CO pressure show that the reaction rate accelerates with decreasing CO pressure.


Pea (Pisum sativum L.) seed albumin extracts show anti-inflammatory effect in the DSS model of mouse colitis.
This study investigates the preventive effects of two pea (Pisum sativum) seed albumin extracts, either in the presence (pea seed extract [PSE]) or absence (albumin fraction from PSE [AF-PSE]) of soluble polysaccharides, in the dextran sodium sulfate (DSS) induced colitis in mice. Male C57BL/6J mice were assigned to five groups: one noncolitic and four colitic. Colitis was induced by incorporating DSS (3.5%) in the drinking water for 4 days, after which DSS was removed. Treated groups received orally PSE (15 g/kg⋅day), or AF-PSE (1.5 g/kg⋅day), or pure soy Bowman-Birk inhibitor (BBI; 50 mg/kg⋅day), starting 2 wk before colitis induction, and maintained for 9 days after. All treated groups showed intestinal anti-inflammatory effect, evidenced by reduced microscopic histological damage in comparison with untreated colitic mice. The treatments ameliorated the colonic mRNA expression of different proinflammatory markers: cytokines, inducible enzymes, metalloproteinases, adhesion molecules, and toll-like receptors, as well as proteins involved in maintaining the epithelial barrier function. Furthermore, the administration of PSE, AF-PSE, or soy BBI restored bacterial counts, partially or totally, to values in healthy mice. PSE and AF-PSE ameliorated DSS-induced damage to mice, their effects being due, at least partially, to the presence of active BBI.


Photocatalytic degradation of organic compounds over combustion-synthesized nano-TiO2.
The photocatalytic degradation of various organics such as phenol, p-nitrophenol, and salicylic acid was carried out with combustion-synthesized nano-TiO2 under UV and solar exposure. Under identical conditions of UV exposure, the initial degradation rate of phenol with combustion-synthesized TiO2 is 2 times higher than the initial degradation rate of phenol with commercial Degussa P-25 TiO2. The intermediates such as catechol (CC) and hydroquinone (HQ) were not detected during the degradation of phenol with combustion-synthesized TiO2, while both the intermediates were detected when phenol was degraded over Degussa P-25. This indicates that the rates of secondary photolysis of CC and HQ occur extremely faster than the rates at which they are formed from phenol and further implies that the primary hydroxylation step is rate limiting for the combustion-synthesized TiO2 aided photodegradation of phenol. The degradation rates of salicylic acid and p-nitrophenol were also investigated, and the rates were higher for combustion-synthesized titania compared to Degussa P-25 TiO2. Superior activity of combustion-synthesized TiO2 toward photodegradation of organic compounds can be attributed to crystallinity, higher surface area, more surface hydroxyl groups, and optical absorption at higher wavelength.


Interrogation of Internal Workings in Microbial Community Assembly: Play a Game through a Behavioral Network?
Increasing evidence shows that the influence of microbiota on biogeochemical cycling, plant development, and human health is executed through a complex network of microbe-microbe interactions. However, characterizing how microbes interact and work together within closely packed and highly heterogeneous microbial ecosystems is extremely challenging. Here, we describe a rule-of-thumb framework for visualizing polymicrobial interactions and extracting general principles that underlie microbial communities. We integrate elements of metabolic ecology, behavioral ecology, and game theory to quantify the interactive strategies by which microbes at any taxonomic level compete for resources and cooperate symbiotically with each other to form and stabilize ecological communities. We show how the framework can chart an omnidirectional landscape of microbial cooperation and competition that may drive various natural processes. This framework can be implemented into genome-wide association studies to unravel the genetic mechanisms underlying microbial interaction networks and their evolutionary consequences along spatiotemporal gradients.IMPORTANCE Identifying general biological rules that underlie the complexity and heterogeneity of microbial communities has proven to be highly challenging. We present a rule-of-thumb framework for studying and characterizing how microbes interact with each other across different taxa to determine community behavior and dynamics. This framework is computationally simple but conceptually meaningful, and it can provide a starting point to generate novel biological hypotheses about microbial interactions and explore internal workings of microbial community assembly in depth.


Database coverage and their use in systematic reviews regarding spinal manipulative therapy: an exploratory study.
Systematic reviews (SRs) of randomized controlled trials (RCTs) are considered one of the most reliable study types. Through a systematic and thorough literature search, researchers aim to collect all research relevant to their purpose. The selection of databases can be challenging and depend on the topic of interest. The Cochrane Handbook suggests searching at least the following three databases: Cochrane Library, MEDLINE, and EMBASE. However, this is not always sufficient for reviews on the musculoskeletal field in general. This study aimed to examine the frequency and choice of databases used by researchers in SRs of spinal manipulative therapy (SMT). Secondly, to analyze the RCTs included in the SRs to determine the optimal combination of databases needed to conduct efficient literature searches for SRs of SMT. SRs investigating the effect of SMT on any patient-reported outcome measure were identified through searches in PubMed and Epistemonikos (all entries till date of search February 25, 2022). For each SR, databases searched and included RCTs were collected. RCTs were searched individually in nine databases (Cochrane Library, MEDLINE/PubMed, EMBASE, Google Scholar, CINAHL, Web of Science, Index to Chiropractic Literature, PEDro, and AMED). Coverage rates were calculated using the number of retrieved RCTs by the database or combinations of databases divided by the total number of RCTs. Eighty-five SRs published met the inclusion criteria, and 442 unique RCTs were retrieved. The most frequently searched database was MEDLINE/PubMed. Cochrane Library had the highest overall coverage rate and contained the third most unique RCTs. While a 100% retrieval was not possible, as 18 RCTs could not be retrieved in any of the nine databases, the combination of Cochrane Library, Google Scholar, and PEDro retrieved all possible RCTs with a combined coverage rate of 95.9%. For SRs on SMT, we recommend using the combination suggested by the Cochrane Handbook of Cochrane Library, MEDLINE/PubMed, Embase, and in addition, PEDro and Index to Chiropractic Literature. Google Scholar might be used additionally as a tool for searching gray literature and quality assurance.


Applicability of Recombinant Laccases From the White-Rot Fungus Obba rivulosa for Mediator-Promoted Oxidation of Biorefinery Lignin at Low pH.
Utilization of lignin-rich side streams has been a focus of intensive studies recently. Combining biocatalytic methods with chemical treatments is a promising approach for sustainable modification of lignocellulosic waste streams. Laccases are catalysts in lignin biodegradation with proven applicability in industrial scale. Laccases directly oxidize lignin phenolic components, and their functional range can be expanded using low-molecular-weight compounds as mediators to include non-phenolic lignin structures. In this work, we studied in detail recombinant laccases from the selectively lignin-degrading white-rot fungus Obba rivulosa for their properties and evaluated their potential as industrial biocatalysts for the modification of wood lignin and lignin-like compounds. We screened and optimized various laccase mediator systems (LMSs) using lignin model compounds and applied the optimized reaction conditions to biorefinery-sourced technical lignin. In the presence of both N-OH-type and phenolic mediators, the O. rivulosa laccases were shown to selectively oxidize lignin in acidic reaction conditions, where a cosolvent is needed to enhance lignin solubility. In comparison to catalytic iron(III)-(2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO) oxidation systems, the syringyl-type lignin units were preferred in mediated biocatalytic oxidation systems.


Interactions of Aspergillus fumigatus and Stenotrophomonas maltophilia in an in vitro Mixed Biofilm Model: Does the Strain Matter?
Introduction: Aspergillus fumigatus (Af) and Stenotrophomonas maltophilia (Sm) are pathogenic microorganisms, which coexist in the respiratory tract of cystic fibrosis (CF) patients. We recently developed an in vitro model of mixed biofilm associating Af ATCC 13073-GFP (Af13073) and Sm ATCC 13637 (Sm13637) and described an antibiosis effect. The present study aim was to assess the antibiosis of Sm on Af using different strains and to analyze the potential synergistic virulence of these strains in an in vivo Galleria mellonella model. Methods: The effect of Sm13637 was evaluated on eight Af strains and the effect of nine Sm strains was evaluated on Af13073. The strains originated from clinical cases (human and animal) and from environment. Fungal and bacterial inocula were simultaneously inoculated to initiate mixed biofilm formation. Fungal growth inhibition was analyzed by qPCR and CLSM and the fungal cell wall modifications by TEM analysis. The virulence of different Sm strains was assessed in association with Af in G. mellonella larvae. Results: All strains of Af and Sm were able to produce single and mixed biofilms. The antibiosis effect of Sm13637 was similar whatever the Af strain tested. On the other hand, the antibiosis effect of Sm strains was bacterial-fitness and strain dependent. One strain (1/9) originated from animal clinical case was never able to induce an antibiosis, even with high bacterial concentration. In the G. mellonella model, co-inoculation with Sm13637 and Af13073 showed synergism since the mortality was 50%, i.e., more than the summed virulence of both. Conclusion: Human clinical strains of Sm yielded in higher antibiosis effect on Af and in a thinner mixed biofilm, probably due to an adaptive effect of these strains. Further research covering Af increased wall thickness in the presence of Sm strains, and its correlation with modified antifungal susceptibility is encouraged in patients with chronic respiratory infections by these 2 microorganisms.


[Multipotent primary bone sarcoma].
The results of clinical and roentgen-morphological studies on 17 cases of multipotent primary sarcoma of the bone are presented. The difficulties of the tumor recognition are due to the fact that it occurs rarely, has no specific clinical-roentgenological picture and shows marked tissue polymorphism. Most frequent are differentiations of the type of osteogenic sarcoma, chondrosarcoma, reticulosarcoma with simultaneous combination with areas having the structure of Ewing's tumor, or extensive fields of undifferentiated primitive tumor cells. Further studies are necessary to decide whether or not this tumor is an independent form among other known skeletal neoplasias.


The Puzzling Unidimensionality of DSM-5 Substance Use Disorder Diagnoses.
There is a perennial expert debate about the criteria to be included or excluded for the DSM diagnoses of substance use dependence. Yet analysts routinely report evidence for the unidimensionality of the resulting checklist. If in fact the checklist is unidimensional, the experts are wrong that the criteria are distinct, so either the experts are mistaken or the reported unidimensionality is spurious. I argue for the latter position, and suggest that the traditional reflexive measurement model is inappropriate for the DSM; a formative measurement model would be a more accurate characterization of the institutional process by which the checklist is created, and a network or causal model would be a more appropriate foundation for a scientifically grounded diagnostic system.


Source Attribution and Risk Assessment of Antimicrobial Resistance.
Source attribution and microbial risk assessment methods have been widely applied for the control of several foodborne pathogens worldwide by identifying (i) the most important pathogen sources and (ii) the risk represented by specific foods and the critical points in these foods' production chains for microbial control. Such evidence has proved crucial for risk managers to identify and prioritize effective food safety and public health strategies. In the context of antimicrobial resistance (AMR) from livestock and pets, the utility of these methods is recognized, but a number of challenges have largely prevented their application and routine use. One key challenge has been to define the hazard in question: Is it the antimicrobial drug use in animals, the antimicrobial-resistant bacteria in animals and foods, or the antimicrobial resistance genes that can be transferred between commensal and pathogenic bacteria in the animal or human gut or in the environment? Other important limitations include the lack of occurrence and transmission data and the lack of evidence to inform dose-response relationships. We present the main principles, available methods, strengths, and weaknesses of source attribution and risk assessment methods, discuss their utility to identify sources and estimate risks of AMR from livestock and pets, and provide an overview of conducted studies. In addition, we discuss remaining challenges and current and future opportunities to improve methods and knowledge of the sources and transmission routes of AMR from animals through food, direct contact, or the environment, including improvements in surveillance and developments in genotypic typing methods.


[Disorders of the integrative brain functions in posttraumatic hydrocephalus].
As many as 110 patients with posttraumatic hydrocephalus in the remote period of mild closed craniocerebral injury underwent routine neuropsychologic evaluation. 80.9% of the patients presented with disorders of higher integrative functions of the brain; of these, 72.7% appeared to get too easily fatigued, their mental processes running to exhaustion too soon. In decreasing rank order are disorders of attention, slowing down of those processes relating to abstract thought, power of apprehension and recent memory (62.7%, 60.9%, and 34.5% respectively). Mnestic disturbances prevailed among patients with moderate severity posttraumatic hydrocephalus (40%) and external hydrocephalus (47.8%). Mild degree hydrocephalus was free from disturbances to higher integrative functions of the brain. The aforesaid attests to the need of the nootropic, vasoactive agents, drugs having resolving and dehydrating effects to be included into the combined treatment of the above patient population.


Relationships between [¹⁸F]-THK5351 Retention and Language Functions in Primary Progressive Aphasia.
There are three distinct subtypes of primary progressive aphasia (PPA): the nonfluent/agrammatic variant (nfvPPA), the semantic variant (svPPA), and the logopenic variant (lvPPA). We sought to characterize the pattern of [¹⁸F]-THK5351 retention across all three subtypes and determine the topography of [¹⁸F]-THK5351 retention correlated with each neurolinguistic score. We enrolled 50 participants, comprising 13 PPA patients (3 nfvPPA, 5 svPPA, and 5 lvPPA) and 37 subjects with normal cognition (NC) who underwent 3.0-tesla magnetic resonance imaging, [¹⁸F]-THK5351 positron-emission tomography scans, and detailed neuropsychological tests. The PPA patients additionally participated in extensive neurolinguistic tests. Voxel-wise and region-of-interest-based analyses were performed to analyze [¹⁸F]-THK5351 retention. The nfvPPA patients exhibited higher [¹⁸F]-THK5351 retention in the the left inferior frontal and precentral gyri. In svPPA patients, [¹⁸F]-THK5351 retention was elevated in the anteroinferior and lateral temporal cortices compared to the NC group (left>right). The lvPPA patients exhibited predominant [¹⁸F]-THK5351 retention in the inferior parietal, lateral temporal, and dorsolateral prefrontal cortices, and the precuneus (left>right). [¹⁸F]-THK5351 retention in the left inferior frontal area was associated with lower fluency scores. Comprehension was correlated with [¹⁸F]-THK5351 retention in the left temporal cortices. Repetition was associated with [¹⁸F]-THK5351 retention in the left inferior parietal and posterior temporal areas, while naming difficulty was correlated with retention in the left fusiform and temporal cortices. The pattern of [¹⁸F]-THK5351 retention was well matched with clinical and radiological findings for each PPA subtype, in agreement with the anatomical and functional location of each language domain.


Inequalities in asthma treatment among children by country of birth and ancestry: a nationwide study in Denmark.
Investigations in several Western countries have reported ethnic differences in asthma prevalence and treatment among children and in some countries these differences are increasing. The aim of this study was to analyse whether there are inequalities in asthma treatment by country of birth and ancestry among children residing in Denmark, and whether this potential association may vary between different household income groups. Data were obtained by linking the Danish Civil Registration System, the Central Taxpayers' Register and the Danish National Prescription Register. the entire population of children in Denmark from 0 to 17 years of age in 2008 (n=1 209 091). Information on asthma treatment was obtained from the National Prescription Register. The analyses included multiple logistic regression models stratified by household income. Compared with ethnic Danes, immigrant children had the lowest OR for redeeming a prescription for asthma medication, both relief (OR 0.37; 95% CIs, 0.20 to 0.68) and preventive (OR 0.37; (0.22 to 0.59)). Similar associations were found among descendant children (OR for relief treatment 0.82 (0.79 to 0.89) and for preventive treatment 0.68 (0.61 to 0.75)). The pattern of the association remained after stratifying for household income. We found that, inequalities that cannot be explained by household income alone exist in treatments to prevent asthma as well as to relieve symptoms in children residing in Denmark, by country of birth and ancestry. The difference between immigrants and descendants may indicate that unfamiliarity with the Danish healthcare system is a contributory cause of the inadequate treatment of asthma.


Separomics applied to the proteomics and peptidomics of low-abundance proteins: Choice of methods and challenges - A review.
The enrichment and isolation of proteins are considered limiting steps in proteomic studies. Identification of proteins whose expression is transient, those that are of low-abundance, and of natural peptides not described in databases, is still a great challenge. Plant extracts are in general complex, and contaminants interfere with the identification of proteins involved in important physiological processes, such as plant defense against pathogens. This review discusses the challenges and strategies of separomics applied to the identification of low-abundance proteins and peptides in plants, especially in plants challenged by pathogens. Separomics is described as a group of methodological strategies for the separation of protein molecules for proteomics. Several tools have been used to remove highly abundant proteins from samples and also non-protein contaminants. The use of chromatographic techniques, the partition of the proteome into subproteomes, and an effort to isolate proteins in their native form have allowed the isolation and identification of rare proteins involved in different processes.


The monoclonal antibody RIB5/1--a useful completion of the MRC OX-3 and OX-6 haplotype specificity.
The haplotype binding pattern of a new mAb RIB5/1 against RT1 class II antigens on different tissues of inbred rat strains was investigated using the indirect POD-technique and a cytofluorometric analysis. RIB5/1 binds to a polymorphic determinant of the RT1 class II glycoprotein. Unlike MRC OX-3 (and OX-6), RIB5/1 reacts in optimal dilution with the RT1 haplotypes dvl, a, av, g, 1 but not with u. Therefore, the mAb pair MRC OX-3 (OX-6) and RIB5/1 is applicable for specific immunohistological purposes in chimaeric models with these RT1 haplotypes.


[Endoluminal, cardiovascular radiotherapy: a new standard].
The treatment for cardiovascular disease, especially the treatment of coronary stenosis, has been continuously improving during the last decades. Routine use of angioplasty was improved by the use of coronary stenting further reducing cardiac morbidity. However, the incidence of restenosis after cardiovascular angioplasty remains high. The restenosis process is mainly explained by neointimal proliferation. Therefore, the utility of ionizing radiation has been systematically investigated in order to reduce proliferation of the neointimal tissue. Radiation therapy turns out to be a very efficient approach in reducing the rate of both de novo lesions as well as of instant restenosis. Recent clinical data from randomized trials confirm the utility of intracoronary radiation therapy and change the treatment standards in interventional cardiology.


Modular peptide binders - development of a predictive technology as alternative for reagent antibodies.
Current biomedical research and diagnostics critically depend on detection agents for specific recognition and quantification of protein molecules. Monoclonal antibodies have been used for this purpose over decades and facilitated numerous biological and biomedical investigations. Recently, however, it has become apparent that many commercial reagent antibodies lack specificity or do not recognize their target at all. Thus, synthetic alternatives are needed whose complex designs are facilitated by multidisciplinary approaches incorporating experimental protein engineering with computational modeling. Here, we review the status of such an engineering endeavor based on the modular armadillo repeat protein scaffold and discuss challenges in its implementation.


Effect of salt, alkali and combined stresses on root system architecture and ion profiling in a diverse panel of oat (Avena spp.).
The co-occurrence of salinisation and alkalisation is quite frequent in problematic soils and poses an immediate threat to food, feed and nutritional security. In the present study, root system architectural traits (RSAs) and ion profiling were evaluated in 21 genotypes of Avena species to understand the effect of salinity-alkalinity stress. The oat genotypes were grown on germination paper and 5-day-old seedlings were transferred to a hydroponic system for up to 30days. These seedlings were subjected to seven treatments: T0, treatment control (Hoagland solution); T1, moderate salinity (50mM); T2, high salinity (100mM); T3, moderate alkalinity (15mM); T4, high alkalinity (30mM); T5, combined moderate salinity-alkalinity (50mM+15mM); and T6, combined high salinity-alkalinity (100mM and 30mM) by using NaCl+Na2SO4 (saline) and NaHCO3+Na2CO3 (alkaline) salts equivalently. The root traits, such as total root area (TRA), total root length (TRL), total root diameter (TRD), total root volume (TRV), root tips (RT), root segments (RS), root fork (RF) and root biomass (RB) were found to be statistically significant (P<0.01) among the different genotypes, treatments and their interactions. Na+ and K+ content analysis in root and shoot tissues revealed the ion homeostasis capacity of different Avena accessions under stress treatments. Principal component analysis (PCA) covered almost 83.0% of genetic variation and revealed that the sharing of TRA, RT, RS and RF traits was significantly high. Biplot analysis showed a highly significant correlation matrix (P<0.01) between the pairs of RT and RS, TRL and RS, and RT and RF. Based on PCA ranking and relative value for stress tolerance, IG-20-1183, IG-20-894, IG-20-718 and IG-20-425 expressed tolerance to salinity (T2), IG-20-425 (alkalinity; T4) and IG-20-1183, IG-20-894 and IG-20-1004 were tolerant to salt-alkali treatment (T6). Multi-trait stability index (MTSI) analysis identified three stable oat genotypes (IG-20-714, IG-20-894 and IG-20-425) under multiple environments and these lines can be used in salinity-alkalinity affected areas after yield trials or as donor lines for combined stresses in future breeding programs.


Centrally-mediated cardiovascular effects of 5-hydroxytryptophan in MAO-inhibited dogs: modification by autonomic antagonists.
In MAO-inhibited dogs, 5-hydroxytryptophan (5-HTP), either 5 or 10 mg/kg i.v., caused hypotension with variable effects on heart rate. Reflex responses to bilateral carotid artery occlusion (BCO) were greatly inhibited by 5-HTP. The hypotensive effects were markedly inhibited by cerebral and extracerebral decarboxylase inhibition with RO 4-4602 and the inhibition of BCO was delayed. Selective extracerebral decarboxylase inhibitation with MK 486 did not prevent either the hypotensive action of 5-HTP or the effect on BCO, although the amino acid now consistently caused bradycardia. Left ventricular pressure and dP/dt were reduced, but cardiac output was maintained by an increase in stroke volume. Hypotension was due predominantly to decreased peripheral resistance. The hypotensive action of 5-HTP was abolished or greatly attenuated after pretreatment with either yohimbine or methysergide, but was unaffected by haloperidol. These results indicate that cerebral decarboxylation and formation of 5-HT are responsible for the hypotension after 5-HTP in MAO inhibited dogs.


Fecal Microbiota Transplantation: The Evolving Risk Landscape.
Fecal microbiota transplantation (FMT) has been recommended in clinical guidelines for the treatment of recurrent Clostridioides difficile infection (CDI). However, it is considered investigational by most regulatory agencies. As the adoption of FMT has increased from a small group of CDI experts alone to more widespread use, there has been a corresponding increase in concern regarding potential risk. FMT is largely considered a safe procedure although risks described range from mild gastrointestinal symptoms to serious infection. Currently, there is variability in how "FMT" is characterized specifically regarding testing approach, which, in turn, impacts the risk profile. This has been highlighted by the rare cases of multidrug-resistant organisms, Shiga toxin-producing Escherichia and enteropathogenic E. coli, recently reported, where these organisms were not screened. These cases have prompted additional screening mandates from the US Food and Drug Administration (FDA), which has maintained its policy of enforcement discretion for the use of FMT for CDI not responding to standard therapy. Here, we examine the evolving risk landscape of FMT.


Untargeted lipidomic analysis in chronic obstructive pulmonary disease. Uncovering sphingolipids.
Cigarette smoke is the major risk factor in the development of chronic obstructive pulmonary disease (COPD). Lipidomics is a novel and emerging research field that may provide new insights in the origins of chronic inflammatory diseases, such as COPD. To investigate whether expression of the sputum lipidome is affected by COPD or cigarette smoking. Lipid expression was investigated with liquid chromatography and high-resolution quadrupole time-of-flight mass spectrometry in induced sputum comparing smokers with and without COPD, and never-smokers. Changes in lipid expression after 2-month smoking cessation were investigated in smokers with and without COPD. More than 1,500 lipid compounds were identified in sputum. The class of sphingolipids was significantly higher expressed in smokers with COPD than in smokers without COPD. At single compound level, 168 sphingolipids, 36 phosphatidylethanolamine lipids, and 5 tobacco-related compounds were significantly higher expressed in smokers with COPD compared with smokers without COPD. The 13 lipids with a high fold change between smokers with and without COPD showed high correlations with lower lung function and inflammation in sputum. Twenty (glyco)sphingolipids and six tobacco-related compounds were higher expressed in smokers without COPD compared with never-smokers. Two-month smoking cessation reduced expression of 26 sphingolipids in smokers with and without COPD. Expression of lipids from the sphingolipid pathway is higher in smokers with COPD compared with smokers without COPD. Considering their potential biologic properties, they may play a role in the pathogenesis of COPD.


Predicting cardiomyopathic phenotypes by altering Ca2+ affinity of cardiac troponin C.
Cardiac diseases associated with mutations in troponin subunits include hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), and restrictive cardiomyopathy (RCM). Altered calcium handling in these diseases is evidenced by changes in the Ca(2+) sensitivity of contraction. Mutations in the Ca(2+) sensor, troponin C (TnC), were generated to increase/decrease the Ca(2+) sensitivity of cardiac skinned fibers to create the characteristic effects of DCM, HCM, and RCM. We also used a reconstituted assay to determine the mutation effects on ATPase activation and inhibition. One mutant (A23Q) was found with HCM-like properties (increased Ca(2+) sensitivity of force and normal levels of ATPase inhibition). Three mutants (S37G, V44Q, and L48Q) were identified with RCM-like properties (a large increase in Ca(2+) sensitivity, partial loss of ATPase inhibition, and increased basal force). Two mutations were identified (E40A and I61Q) with DCM properties (decreased Ca(2+) sensitivity, maximal force recovery, and activation of the ATPase at high [Ca(2+)]). Steady-state fluorescence was utilized to assess Ca(2+) affinity in isolated cardiac (c)TnCs containing F27W and did not necessarily mirror the fiber Ca(2+) sensitivity. Circular dichroism of mutant cTnCs revealed a trend where increased alpha-helical content correlated with increased Ca(2+) sensitivity in skinned fibers and vice versa. The main findings from this study were as follows: 1) cTnC mutants demonstrated distinct functional phenotypes reminiscent of bona fide HCM, RCM, and DCM mutations; 2) a region in cTnC associated with increased Ca(2+) sensitivity in skinned fibers was identified; and 3) the F27W reporter mutation affected Ca(2+) sensitivity, maximal force, and ATPase activation of some mutants.


Loss of IFN gamma receptor is an independent prognostic factor in ovarian cancer.
There is evidence that IFN gamma plays an important role in ovarian cancer development. IFN gamma produces numerous antitumor effects and it may be evasion of these effects which allows tumor progression. We postulate that genetic instability in tumor cells may lead to modulation of expression of the IFN gamma receptor, thus leading to altered tumor biology and patient prognosis. This hypothesis would support the theory of immunoediting in ovarian cancer. Using tissue microarray technology of 339 primary ovarian cancers, the expression of IFN gamma receptor was assessed immunohistochemically. Coupled to a comprehensive database of clinicopathologic variables, its effect on these factors was studied. Tumors expressing high levels of IFN gamma receptor had significantly improved survival (P=0.017) compared with tumors expressing low levels of the receptor; this was also seen with complete receptor loss (P=0.014). Factors shown to predict prognosis independently of each other were the following: age, International Federation of Gynecologists and Obstetricians stage, and the absence of macroscopic disease after surgery. The level of IFN gamma receptor expression and complete receptor loss were independently predictive of prognosis on multivariate analysis. There was no correlation between receptor status and any of the standard clinicopathologic variables. Loss of IFN gamma receptor independently predicts poor prognosis in ovarian cancer. Loss of receptor expression may be responsible for the limited success in the therapeutic use of IFN gamma in ovarian cancer trials and highlights a subgroup of high expressing IFN gamma receptor tumors which are more likely to be susceptible to such treatments.


Diversity and evolution of the Cydia pomonella granulovirus.
Eight new field isolates of Cydia pomonella granulovirus (CpGV) originating in Iran and Georgia and one English CpGV isolate were analysed for restriction fragment length polymorphisms (RFLPs) and by partial genome amplification and sequencing. According to the observed RFLPs, most of the predominant genotypes of these isolates could be assigned to those present in previously found isolates originating from Mexico (CpGV-M), England (CpGV-E) and Russia (CpGV-R). We suggest that these isolates should be designated genome A, B and C types, respectively. A fourth genome type was identified in three isolates and is designated D type. The isolates with A, B and D type genomes contained four open reading frames (ORFs) (ORF63-ORF66) not present in C type genomes. The lack of these ORFs in other granuloviruses suggests that the C type genome is evolutionarily ancestral to the other genome types. The B and D type genomes contained an additional insertion of a non-protein coding region of 0.7 kb, which was at different genome locations. Analysis of the partial gene sequences of late expression factor 8 (lef-8), lef-9 and polyhedrin/granulin (polh/gran) genes revealed single nucleotide polymorphisms (SNPs) that corresponded to the RFLP types. Phylogenetic analyses based on these SNPs corroborated the proposed ancestry of the C type genome. C type viruses were also less virulent to neonate codling moth larvae than the other virus types. In conclusion, the known diversity of CpGV isolates can be described by four major genome types, which appear to exist in different isolates as genotype mixtures.


Health-related outcomes of importance to patients with Takayasu's arteritis.
The need to include patients' perspectives as key outcomes in clinical trials is widely accepted. No disease-specific patient-reported outcomes have been developed in Takayasu's arteritis. This project was designed to identify outcomes of importance to patients with Takayasu's arteritis during active disease and remission across 2 different cultures. Patients with Takayasu's arteritis from the US and Turkey were recruited to participate in semi-structured, one-on-one interviews or focus groups. The interviews and group sessions were recorded, transcribed, and entered into an Nvivo database. A line-by-line review of narrative data was used to develop themes describing the impact of Takayasu's arteritis on patients' life. US Patients were invited to freelist terms that they associated with disease states (active disease and remission). The Smith's Salience Index (SSI) was used to identify the most salient terms. Results. A total of 31 patients with Takayasu's arteritis participated in this study. Interviews and focus groups identified pain, fatigue, and emotional impact as common themes. Outcomes did not differ between the 2 countries. The most salient terms identified through freelisting were pain/discomfort and fatigue/low energy levels (SSI=0.56 and 0.33, respectively) during active disease and pain/discomfort and emotional impact (SSI=0.51 and 0.37, respectively) during remission. Patients with Takayasu's arteritis report a range of disease-specific symptoms across different cultures and disease states that are generally not specifically captured by generic patient-reported outcome tools currently used in research. Identifying disease-specific outcomes would advance clinical trials methodology to best capture the full spectrum of disease activity in Takayasu's arteritis.


Health-Related Quality of Life and Its Determinants Among Hypertensive Patients in Rural Southwest Nigeria.
The health-related quality of life (HRQOL) of patients with hypertension is of particular concern to stakeholders because hypertension is a chronic disease. To determine the pattern of HRQOL and its determinants among hypertensive patients in rural Southwestern Nigeria. Ahospital-based crosssectional survey was conducted between June and October 2019 on 390 consented adult patients with hypertension on follow-up using systematic sampling technique. Respondents'blood pressure (BP), body mass index (BMI), waist circumference (WC), waist-hip ratio (W-HR) and other measurements were taken. Relevant data on socio-demographics and lifestyle behaviours were collected using an interviewer-administered questionnaire which incorporated validated instrument for assessment of HRQOL score using SF-12 survey. The data were analyzed using SPSS (version 21). Binary logistic regression was used to identify the determinants of HRQOL. The mean physical component summary (42.0 ± 8.44), mental component summary (40.6 ± 8.11), and mean overall quality of life score (41.3 ± 8.17) were all below the normal score of 50 ±10. However, of all the eight domains, the role emotional was the highest (47.61 ± 14.97). The odds of having overall good HRQOL were better with respondents with female gender (AOR=1.99, 95% CI: 1.05-3.88), tertiary education (AOR=3.22, 95% CI: 1.63-7.20), and controlled BP (AOR=3.01,95% CI: 1.04-15.79). The mean HRQOLscore of the respondents was below the mean score. The findings of this study would assist stakeholders in decision-making towards improved management of patients with hypertension.


Porcine UL16-binding protein 1 expressed on the surface of endothelial cells triggers human NK cytotoxicity through NKG2D.
Cellular rejection mechanisms, including NK cells, remain a hurdle for successful pig-to-human xenotransplantation. Human anti-pig NK cytotoxicity depends on the activating receptor NKG2D. Porcine UL16-binding protein 1 (pULBP1) and porcine MHC class I chain-related protein 2 (pMIC2) are homologues of the human NKG2D ligands ULBP 1-4 and MICA and B, respectively. Although transcribed in porcine endothelial cells (pEC), it is not known whether pULBP1 and pMIC2 act as functional ligands for human NKG2D. In this study, surface protein expression of pULBP1 was demonstrated by flow cytometry using a novel pULBP1-specific polyclonal Ab and by cellular ELISA using NKG2D-Fc fusion protein. Reciprocally, pULBP1-Fc bound to primary human NK cells, whereas pMIC2-Fc did not. Transient and stable down-regulation of pULBP1 mRNA in pEC using short-interfering RNA oligonucleotide duplexes and short hairpin RNA, respectively, resulted in a partial inhibition of xenogeneic NK cytotoxicity through NKG2D in (51)Cr release assays. In contrast, down-regulation of pMIC2 mRNA did not inhibit NK cytotoxicity. Human NK cytotoxicity against pEC mediated by freshly isolated or IL-2-activated NK cells through NKG2D was completely blocked using anti-pULBP1 polyclonal Ab. In conclusion, this study suggests that pULBP1 is the predominant, if not only, functional porcine ligand for human NKG2D. Thus, the elimination of pULBP1 on porcine tissues represents an attractive target to protect porcine xenografts from human NK cytotoxicity.


[Occupational factors in parents and morbidity in their children].
Nonspecific resistance and the incidence of allergic diseases were studied in children living in areas with varying ambient air pollution, whose parents were exposed before conception to deleterious occupational factors at chemical and petrochemical enterprises. Nonspecific resistance was found to reduce drastically in children of female workers of chemical and petrochemical enterprises. In the children of female workers of the above enterprises, the incidence of allergic diseases was significantly higher (p < 0.01) than that in the control group. The findings suggest that environmental factors (ambient air pollution, occupational harms in parents) exert a substantial influence on a rise in the incidence of allergic diseases in children.


Diagnostic value of surgical testicular biopsy after therapy for acute lymphocytic leukemia.
Inasmuch as several cooperative research groups currently require routine testicular biopsy in boys with acute lymphocytic leukemia before cessation of therapy, we performed bilateral open wedge biopsies in 38 boys aged 4 1/2 to 18 years, all of whom were completing 3 years of therapy, were apparently in complete remission, and had palpably normal testes. Biopsy specimens from two other patients were examined during therapy for specific clinical indications. Among the 40 patients, one biopsy revealed leukemia bilaterally; 39 patients had negative biopsy results, three of whom had overt, biopsy-proved testicular relapse 6 weeks to 17 months after normal biopsy. All four patients with testicular relapse are in complete remission six to 50 months after testicular irradiation. We conclude that random sampling during routine testicular biopsies may fail to demonstrate occult leukemia, and that the high rate of complete remissions following retrieval therapy after overt testicular relapse makes routine biopsies before cessation of therapy unnecessary.


Anxiety Sensitivity Index (ASI-3) subscales predict unique variance in anxiety and depressive symptoms.
Anxiety sensitivity (AS) has been implicated in the development and maintenance of a range of mental health problems. The development of the Anxiety Sensitivity Index - 3, a psychometrically sound index of AS, has provided the opportunity to better understand how the lower-order factors of AS - physical, psychological, and social concerns - are associated with unique forms of psychopathology. The present study investigated these associations among 85 treatment-seeking adults with high AS. Participants completed measures of AS, anxiety, and depression. Multiple regression analyses controlling for other emotional disorder symptoms revealed unique associations between AS subscales and certain types of psychopathology. Only physical concerns predicted unique variance in panic, only cognitive concerns predicted unique variance in depressive symptoms, and social anxiety was predicted by only social concerns. Findings emphasize the importance of considering the multidimensional nature of AS in understanding its role in anxiety and depression and their treatment.


Approximation of nonlinear systems with radial basis function neural networks.
A technique for approximating a continuous function of n variables with a radial basis function (RBF) neural network is presented. The method uses an n-dimensional raised-cosine type of RBF that is smooth, yet has compact support. The RBF network coefficients are low-order polynomial functions of the input. A simple computational procedure is presented which significantly reduces the network training and evaluation time. Storage space is also reduced by allowing for a nonuniform grid of points about which the RBFs are centered. The network output is shown to be continuous and have a continuous first derivative. When the network is used to approximate a nonlinear dynamic system, the resulting system is bounded-input bounded-output stable. For the special case of a linear system, the RBF network representation is exact on the domain over which it is defined, and it is optimal in terms of the number of distinct storage parameters required. Several examples are presented which illustrate the effectiveness of this technique.


Two biochemically distinct lipophosphoglycans from Leishmania braziliensis and Leishmania infantum trigger different innate immune responses in murine macrophages.
The dominant, cell surface lipophosphoglycan (LPG) of Leishmania is a multifunctional molecule involved in the interaction with vertebrate and invertebrate hosts. Although the role of LPG on infection has been extensively studied, it is not known if LPG interspecies variations contribute to the different immunopathologies of leishmaniases. To investigate the issue of interspecies polymorphisms, two Leishmania species from the New World that express structural variations of side chains of LPG repeat units were examined. In this context, the procyclic form of L. braziliensis LPG (strain M2903), is devoid of side chains, while the L. infantum LPG (strain BH46) has up to three glucoses residues in the repeat units. Mice peritoneal macrophages from Balb/c, C57BL/6 and knock-out (TLR2 -/-, TLR4 -/-) were primed with IFN-γ and stimulated with purified LPG from both species. Nitric oxide and cytokine production, MAPKs (ERK, p38 and JNK) and NF-kB activation were evaluated. Macrophages stimulated with L. braziliensis LPG, had a higher TNF-α, IL-1β, IL-6 and NO production than those stimulated with that of L. infantum. Furthermore, the LPGs from the two species resulted in differential kinetics of signaling via MAPK activation. L. infantum LPG exhibited a gradual activation profile, whereas L. braziliensis LPG showed a sharp but transient activation. L. braziliensis LPG was able to activate NF-kB. These data suggest that two biochemically distinct LPGs were able to differentially modulate macrophage functions.


Peptide-coated polyurethane material reduces wound infection and inflammation.
There is an urgent need for treatments that not only reduce bacterial infection that occurs during wounding but that also target the accompanying excessive inflammatory response. TCP-25, a thrombin-derived antibacterial peptide, scavenges toll-like receptor agonists such as endotoxins and lipoteichoic acid and prevents toll-like receptor-4 dimerization to reduce infection-related inflammation in vivo. Using a combination of biophysical, cellular, and microbiological assays followed by experimental studies in mouse and pig models, we show that TCP-25, when delivered from a polyurethane (PU) material, exerts anti-infective and anti-inflammatory effects in vitro and in vivo. Specifically, TCP-25 killed the common wound pathogens, Pseudomonas aeruginosa and Staphylococcus aureus, in both in vitro and in vivo assays. Furthermore, after its release from the PU material, the peptide retained its capacity to induce its helical conformation upon endotoxin interaction, yielding reduced activation of NF-κB in THP-1 reporter cells, and diminished accumulation of inflammatory cells and subsequent release of IL-6 and TNF-α in subcutaneous implant models in vivo. Moreover, in a porcine partial thickness wound infection model, TCP-25 treated infection with S. aureus, and reduced the concomitant inflammatory response. Taken together, these findings demonstrate a combined antibacterial and anti-inflammatory effect of TCP-25 delivered from PU in vitro, and in mouse and porcine in vivo models of localized infection-inflammation. STATEMENT OF SIGNIFICANCE: Local wound infections may result in systemic complications and can be difficult to treat due to increasing antimicrobial resistance. Surgical site infections and biomaterial-related infections present a major challenge for hospitals. In recent years, various antimicrobial coatings have been developed for infection prevention and current concepts focus on various matrices with added anti-infective components, including various antibiotics and antiseptics. We have developed a dual action wound dressing concept where the host defense peptide TCP-25, when delivered from a PU material, targets both bacterial infection and the accompanying inflammation. TCP-25 PU showed efficacy in in vitro and experimental wound models in mouse and minipigs.


Prologue: 2009 Group for Research and Assessment of Psoriasis and Psoriatic Arthritis (GRAPPA).
The 2009 Annual Meeting of the Group for Research and Assessment of Psoriasis and Psoriatic Arthritis (GRAPPA) was held in June 2009 in Stockholm, Sweden, and was attended by rheumatologists, dermatologists, biopharmaceutical company representatives, and patient groups. A primary goal of GRAPPA is to foster outreach and interdisciplinary communication between the fields of rheumatology and dermatology. Several members attended an adjacent meeting of the International Federation of Psoriasis Associations; reports were also provided of recent meetings of the American Academy of Dermatology and the Assessment of SpondyloArthritis (ASAS) working group. In a training session of the GRAPPA meeting, members served as faculty while rheumatology fellows and dermatology residents presented original research work. In one module of the meeting, several response measures were discussed. In another module, discussions were held on the need for dermatologists to be able to diagnose psoriatic arthritis (PsA) among their psoriasis patients; several PsA screening questionnaires were presented, and progress was reported on developing online training videos as an aid to educate clinicians in their diagnoses. Other topics for discussion at the GRAPPA meeting included presentations on genetic associations with PsA and on comorbidities in patients with PsA. Current and future research projects also were outlined.


Adult Otogenic Meningitis in the Pneumococcal Conjugated Vaccines Era.
Introduction Streptococcus pneumoniae is a major pathogen of otogenic meningitis (OgM), the most commonly reported intra-cranial complication of otitis media (OM). Objectives To study the changes in adult OgM patients in the pneumococcal conjugated vaccines (PCVs) era. Methods Retrospective cohort of adults presenting with concurrent OM and meningitis in a secondary medical care center between 2005 and 2015. Data collected included demographics, OM-related symptoms, cerebrospinal fluid (CSF) and ear culture results, pre- and during hospitalization antibiotic treatment, imaging findings, and complications. We compared the pre-PCV years (2005-2009) with the post-PCV years (2010-2015). Outcomes were 1) incidence of all-cause adult OgM from the total meningitis cases; 2) impact of PCVs on the clinical presentation of OgM and bacteriology. Results Otogenic meningitis was diagnosed in 26 out of 45 (58%) cases of all-cause meningitis admissions. Of those, 10 (38%) were male, with a mean age of 62.4 years old. Ear-related signs and symptoms were documented in 70% of the patients, and OgM was diagnosed following imaging studies in 6 out of 26 (23%) patients. All of the patients received intravenous antibiotic therapy, and 7 (27%) patients required surgical interventions: 6 required myringotomy and 1 required mastoidectomy. There were 12 (46%) patients in the pre-PCV years and 14 (56%) in the post-PCV years. The positive pneumococcal CSF and ear culture rates remained high and unchanged (∼ 75%). There were no significant changes in the clinical presentation or mortality rates. Conclusion Micro-otoscopy should be included in the routine work-up of any suspected adult meningitis, because OgM is underdiagnosed. Unlike their impact on pediatric otitis media, PCVs did not change the epidemiology and bacteriology of OgM.


A scoping review of crisis teams managing dementia in older people.
Research on crisis teams for older adults with dementia is limited. This scoping review aimed to 1) conduct a systematic literature review reporting on the effectiveness of crisis interventions for older people with dementia and 2) conduct a scoping survey with dementia crisis teams mapping services across England to understand operational procedures and identify what is currently occurring in practice. For the systematic literature review, included studies were graded using the Critical Appraisal Skills Programme checklist. For the scoping survey, Trusts across England were contacted and relevant services were identified that work with people with dementia experiencing a mental health crisis. The systematic literature review demonstrated limited evidence in support of crisis teams reducing the rate of hospital admissions, and despite the increase in number of studies, methodological limitations remain. For the scoping review, only half (51.8%) of the teams had a care pathway to manage crises and the primary need for referral was behavioral or psychological factors. Evidence in the literature for the effectiveness of crisis teams for older adults with dementia remains limited. Being mainly cohort designs can make it difficult to evaluate the effectiveness of the intervention. In practice, it appears that the pathway for care managing crisis for people with dementia varies widely across services in England. There was a wide range of names given to the provision of teams managing crisis for people with dementia, which may reflect the differences in the setup and procedures of the service. To provide evidence on crisis intervention teams, a comprehensive protocol is required to deliver a standardized care pathway and measurable intervention as part of a large-scale evaluation of effectiveness.


Is there a twelfth protein-coding gene in the genome of influenza A? A selection-based approach to the detection of overlapping genes in closely related sequences.
Protein-coding genes often contain long overlapping open-reading frames (ORFs), which may or may not be functional. Current methods that utilize the signature of purifying selection to detect functional overlapping genes are limited to the analysis of sequences from divergent species, thus rendering them inapplicable to genes found only in closely related sequences. Here, we present a method for the detection of selection signatures on overlapping reading frames by using closely related sequences, and apply the method to several known overlapping genes, and to an overlapping ORF on the negative strand of segment 8 of influenza A virus (NEG8), for which the suggestion has been made that it is functional. We find no evidence that NEG8 is under selection, suggesting that the intact reading frame might be non-functional, although we cannot fully exclude the possibility that the method is not sensitive enough to detect the signature of selection acting on this gene. We present the limitations of the method using known overlapping genes and suggest several approaches to improve it in future studies. Finally, we examine alternative explanations for the sequence conservation of NEG8 in the absence of selection. We show that overlap type and genomic context affect the conservation of intact overlapping ORFs and should therefore be considered in any attempt of estimating the signature of selection in overlapping genes.


Effectiveness of Food-Based Intervention to Improve the Linear Growth of Children under Five: A Systematic Review and Meta-Analysis.
Food-based interventions to improve linear growth are most often applied in low- and middle-income countries. However, not all food interventions have been proven to be effective in promoting linear growth. This study aimed to assess the impact and effectiveness of food interventions for improving linear growth in children under five years old. This study was conducted by following the PRISMA guidelines and the data were extracted and presented following the PRISMA recommendations. Studies were identified through a literature search of the SCOPUS, Web of Science, PubMed, ScienceDirect, and ProQuest databases from 2000 to 2022. Only randomized control studies were included in this review based on the inclusion and exclusion criteria. Out of 1125 studies identified, a total of 15 studies were included in this systematic review and meta-analysis. The review result indicated that food-based intervention can help to improve linear growth (MD: 0.20, 95% CI: 0.04 to 0.35, p = 0.01) among children under five. However, there was no significant difference in changes in underweight status (MD: 0.25; CI: -0.15 to 0.64; p = 0.22) and wasting status (MD: 0.09; CI: -0.02 to 0.20; p = 0.12) between the intervention and control groups. Overall, food-based interventions were found to be helpful for improving children's linear growth.


Limitations to photosynthesis in leaves of wheat plants infected by Pyricularia oryzae.
Blast, caused by Pyricularia oryzae, has become an economically important disease in wheat in Brazil, but little effort has been devoted to understanding the wheat-P. oryzae interaction. This study was intended to determine the effects of P. oryzae infection on the photosynthetic process in wheat plants using a susceptible (BR 18) and a partially resistant cultivar (BRS 229). It was found that the net carbon assimilation rate (A), stomatal conductance (gs), and transpiration rate were dramatically reduced in both cultivars due to P. oryzae infection but to a lesser degree in BRS 229. Photosynthesis was impaired in asymptomatic leaf tissues, indicating that blast severity is not an acceptable indicator for predicting P. oryzae-induced reductions in A. The proportionally larger decreases in A than in gs, in parallel with increases in internal CO2 concentration (Ci), suggest that the lower influx of CO2 into the diseased leaves caused by stomatal closure was not a prominent factor associated with the reduction in A. Additional support for this conclusion comes from the nonsignificant correlation between A and gs, the negative correlation between A and Ci and the positive correlation between blast severity and Ci. Both the maximum rate of carboxylation and the maximum rate of electron transport were dramatically depressed at advanced stages of P. oryzae infection, mainly in BR 18, although the reduction in A was not closely related to the decrease in the electron transport rate. In conclusion, biochemical limitations likely related to the reduced activity of Rubisco, rather than diffusive limitations, were the main factor associated with decreases in A during the infection process of P. oryzae on wheat leaves.


MUC20 suppresses the hepatocyte growth factor-induced Grb2-Ras pathway by binding to a multifunctional docking site of met.
A cDNA encoding a novel mucin protein, MUC20, was isolated as a gene that is up-regulated in the renal tissues of patients with immunoglobulin A nephropathy. We demonstrate here that the C terminus of MUC20 associates with the multifunctional docking site of Met without ligand activation, preventing Grb2 recruitment to Met and thus attenuating hepatocyte growth factor (HGF)-induced transient extracellular signal-regulated kinase-1 and -2 activation. Production of MUC20 reduced HGF-induced matrix metalloproteinase expression and proliferation, which require the Grb2-Ras pathway, whereas cell scattering, branching morphogenesis, and survival via the Gab1/phosphatidylinositol 3-kinase (PI3K) pathways was not affected. Thus, MUC20 reduces HGF-induced activation of the Grb2-Ras pathway but not the Gab1/PI3K pathways. We further demonstrate that the cytoplasmic domain of MUC20 has the ability to oligomerize and that the oligomerization augments its affinity for Met. Taken together, these results suggest that MUC20 is a novel regulator of the Met signaling cascade which has a role in suppression of the Grb2-Ras pathway.


Uterine myxoid leiomyosarcoma with tumor embolism extending into the right atrium.
Uterine myxoid leiomyosarcoma (MLMS) is an extremely rare variant of uterine leiomyosarcoma; only 56 cases were reported from 1982 to 2013. Uterine MLMS is characterized by a myxoid appearance and highly malignant behavior. We herein report a case involving a 65-year-old woman with uterine MLMS with a large tumor embolism that reached the right atrium. A total abdominal hysterectomy, bilateral salpingooophorectomy, and tumor embolism resection with the use of a heart-lung machine were performed. Epirubicin-ifosfamide chemotherapy in the adjuvant setting led to reductions in both the tumor emboli and peritoneal dissemination. The patient retained a good quality of life for 10 months after the initial surgery. She then developed progressive disease despite treatment with pazopanib. She died of her disease 14 months after the initial surgery. Although complete surgical resection of the tumor is desirable, tumor reduction surgery followed by adjuvant chemotherapy might help to retain a good quality of life. This is the first reported case of a primary uterine MLMS with tumor emboli.


The right to traditional, complementary, and alternative health care.
State parties to human rights conventions and declarations are often faced with the seemingly contradictory problem of having an obligation to protect people from harmful practices while also having an obligation to enable access to culturally appropriate effective healing. As people increasingly migrate across the globe, previous distinctions between 'traditional' and 'complementary and alternative medicine' practices are being transcended. There are connections across transnational healing pathways that link local, national, and global movements of people and knowledge. This paper contributes to the development of the concept and practice of the right to health in all its forms, exploring the right to traditional, complementary, and alternative health (R2TCAH) across different contexts. The paper draws on four settings - England, South Africa, Kenya, and Jordan - and is based on key informant interviews and a literature review undertaken in 2010, and updated in 2013. The paper begins by reviewing the international legal context for the right to health. It then considers legal and professional regulations from the global north and south. Additional research is needed to establish the legal basis, compare regulatory frameworks, and explore patient and provider perspectives of regulation. This leads to being able to make recommendations on how to balance protection from harm and the obligation to ensure culturally appropriate services. Such an exploration must also challenge Western theories of human rights. Key concepts, such as individual harm, consent, and respect of the autonomy of the individual already established and recognised in international health law, could be adopted in the development of a template for future comparative research. Exploration of the normative content of the right to health in all its forms will contribute to supporting traditional, complementary, and alternative health service users and providers in terms of access to information, non-discrimination, clarification of state obligations, and accountability.


Adaptive multilevel splitting: Historical perspective and recent results.
This article first presents a short historical perpective of the importance splitting approach to simulate and estimate rare events, with a detailed description of several variants. We then give an account of recent theoretical results on these algorithms, including a central limit theorem for Adaptive Multilevel Splitting (AMS). Considering the asymptotic variance in the latter, the choice of the importance function, called the reaction coordinate in molecular dynamics, is also discussed. Finally, we briefly mention some worthwhile applications of AMS in various domains.


Campylobacter jejuni contains two fur homologs: characterization of iron-responsive regulation of peroxide stress defense genes by the PerR repressor.
Expression of the peroxide stress genes alkyl hydroperoxide reductase (ahpC) and catalase (katA) of the microaerophile Campylobacter jejuni is repressed by iron. Whereas iron repression in gram-negative bacteria is usually carried out by the Fur protein, previous work showed that this is not the case in C. jejuni, as these genes are still iron repressed in a C. jejuni fur mutant. An open reading frame encoding a Fur homolog (designated PerR for "peroxide stress regulator") was identified in the genome sequence of C. jejuni. The perR gene was disrupted by a kanamycin resistance cassette in C. jejuni wild-type and fur mutant strains. Subsequent characterization of the C. jejuni perR mutants showed derepressed expression of both AhpC and KatA at a much higher level than that obtained by iron limitation, suggesting that expression of these genes is controlled by other regulatory factors in addition to the iron level. Other iron-regulated proteins were not affected by the perR mutation. The fur perR double mutant showed derepressed expression of known iron-repressed genes. Further phenotypic analysis of the perR mutant, fur mutant, and the fur perR double mutant showed that the perR mutation made C. jejuni hyperresistant to peroxide stress caused by hydrogen peroxide and cumene hydroperoxide, a finding consistent with the high levels of KatA and AhpC expression, and showed that these enzymes were functional. Quantitative analysis of KatA expression showed that both the perR mutation and the fur mutation had profound effects on catalase activity, suggesting additional non-iron-dependent regulation of KatA and, by inference, AhpC. The PerR protein is a functional but nonhomologous substitution for the OxyR protein, which regulates peroxide stress genes in other gram-negative bacteria. Regulation of peroxide stress genes by a Fur homolog has recently been described for the gram-positive bacterium Bacillus subtilis. C. jejuni is the first gram-negative bacterium where non-OxyR regulation of peroxide stress genes has been described and characterized.


Socioeconomic disparities in suicide: Causation or confounding?
Despite an overall reduction in suicide, educational disparities in suicide have not decreased over the last decade. The mechanisms behind educational disparities in suicide, however, remain unclear: low educational status may increase the risk of suicide ("causation") or low educational status and suicide may share confounders. This paper assesses whether educational disparities in suicide (EDS) are more likely to be due to causation. The DEMETRIQ study collected and harmonized register-based data on mortality follow-up from forty population censuses from twelve European populations. More than 102,000 suicides were registered over 392 million person-years. Three analyses were carried out. First, we applied an instrumental variable approach that exploits changes in the legislation on compulsory educational age to instrument educational status. Second, we analyzed EDS by age under the hypothesis that increasing EDS over the life cycle supports causation. Finally, we compared EDS in men and women under the assumption that greater EDS in women would support causation. The instrumental variable analysis showed no evidence for causation between higher education and suicide, for men or women. The life-cycle analysis showed that the decrease of educational inequalities in suicide between the baseline 1991 period and the 2001 follow-up period was more pronounced and statistically significant in the first three younger age groups. The gender analysis indicated that EDS were systematic and greater in men than in women: the rate ratio of suicide for men with low level of education (RR = 2.51; 95%CI:2.44-2.58) was higher than the rate ratio in women (RR = 1.32; 95CI%:1.26-1.38). Overall, there was little support for the causation hypothesis, suggesting that the association between education and suicide is confounded. Educational inequalities in suicide should be addressed in early life by early targeting of groups who struggle to complete their education and display higher risk of mental disorder or of mental health vulnerabilities.


Time Series Prediction of Lung Cancer Death Rates on the Basis of SEER Data.
The purpose of this study was to apply different time series analytical techniques to SEER US lung cancer death rate data to develop a best fit model. Three models for yearly time series predictions were built: autoregressive integrated moving average (ARIMA), simple exponential smoothing (SES), and Holt's double expansional smoothing (HDES) models. The three models were built using Python 3.9, on the basis of Anaconda 2022.10. This study used SEER data from 1975 to 2018 and included 545,486 patients with lung cancer. The best parameters for ARIMA are ARIMA (p, d, q) = (0, 2, 2). In addition, the best parameter for SES was α = .995, whereas the best parameters for HDES were α = .4 and β = .9. The HDES was the model that best fit the lung cancer death rate data, with a root mean square error (RMSE) of 132.91. Including monthly diagnoses, death rates, and years in SEER data increases the number of observations for training and test sets, enhancing the performance of time series models. The reliability of the RMSE was based on the mean lung cancer mortality rate. Owing to the high mean lung cancer death rate of 8,405 patients per year, it is acceptable for reliable models to have large RMSEs.


Resolution enhancement through microscopic spatiotemporal control.
Operating at biologically benign conditions, multi-photon fluorescence imaging microscopy has benefitted immensely from recent developments in microscopic resolution enhancement. Fluorescence microscopy continues to be the best choice for experiments on live specimens, however, multi-photon fluorescence imaging often suffers from overlapping fluorescence of typical dyes used in microscopy, limiting its scope. This limitation has been the focus of our research where we show that by making simple modifications to the laser pulse structure, it is possible to resolve these overlapping fluorescence complications. Specifically, by using pairs of femtosecond pulses with variable delay in place of single pulse excitation, we show controlled fluorescence excitation or suppression of one of the fluorophores over the other through wave-packet interferometry. Such an effect prevails even after the fluorophore coherence timescale, which effectively results in a higher spatial resolution. Here we extend the effect of our pulse-pair technique to microscopic axial resolution experiments and show that such pairs of pulses can also 'enhance' axial resolution.


A luteinizing hormone-releasing hormone agonist for the prevention of chemotherapy-induced ovarian follicular loss in rats.
In an attempt to prevent chemotherapy-induced ovarian follicular loss, [D-Leu6, des-Gly10-NH2]-luteinizing hormone-releasing hormone ethylamide (LHRHa) was given subcutaneously to Sprague-Dawley cycling female rats in two daily doses of 2.5 micrograms starting 2 days prior to and concomitant with cyclophosphamide (CTX) (5 mg/kg/day for 21 days). Four groups of female cycling rats (10 in each) received either no treatment, CTX alone, CTX + LHRHa, or LHRHa alone. One ovary from each animal was serially sectioned, stained, and examined for the number and size of follicles. CTX produced a significant reduction in the total number of follicles. The pool of growing follicles (medium to large, greater than 30 microns in diameter) appeared to be vulnerable to the cytotoxic effect of CTX. LHRHa resulted in a significant reduction in the number of medium-to-large follicles and an increase in the number of small follicles. When given in combination with CTX, LHRHa significantly further reduced the number of medium-to-large follicles, significantly increased the number of small follicles, and resulted in an increase in the total number of follicles. Chronic LHRHa treatment resulted in functional deprivation of follicles from gonadotropins, thus halting the process of recruitment from the quiescent pool of primordial follicles into the CTX sensitive pool and thereby preserving the functional potential of the ovary.


The aminoglycoside resistance methyltransferase Sgm impedes RsmF methylation at an adjacent rRNA nucleotide in the ribosomal A site.
Ribosome-targeting antibiotics block protein synthesis by binding at functionally important regions of the bacterial rRNA. Resistance is often conferred by addition of a methyl group at the antibiotic binding site within an rRNA region that is already highly modified with several nucleotide methylations. In bacterial rRNA, each methylation requires its own specific methyltransferase enzyme, and this raises the question as to how an extra methyltransferase conferring antibiotic resistance can be accommodated and how it can gain access to its nucleotide target within a short and functionally crowded stretch of the rRNA sequence. Here, we show that the Sgm methyltransferase confers resistance to 4,6-disubstituted deoxystreptamine aminoglycosides by introducing the 16S rRNA modification m(7)G1405 within the ribosomal A site. This region of Escherichia coli 16S rRNA already contains several methylated nucleotides including m(4)Cm1402 and m(5)C1407. Modification at m(5)C1407 by the methyltransferase RsmF is impeded as Sgm gains access to its adjacent G1405 target on the 30S ribosomal subunit. An Sgm mutant (G135A), which is impaired in S-adenosylmethionine binding and confers lower resistance, is less able to interfere with RsmF methylation on the 30S subunit. The two methylations at 16S rRNA nucleotide m(4)Cm1402 are unaffected by both the wild-type and the mutant versions of Sgm. The data indicate that interplay between resistance methyltransferases and the cell's own indigenous methyltransferases can play an important role in determining resistance levels.


Emotional and Motivational Pain Processing: Current State of Knowledge and Perspectives in Translational Research.
Pain elicits fear and anxiety and promotes escape, avoidance, and adaptive behaviors that are essential for survival. When pain persists, motivational priority and attention shift to pain-related information. Such a shift often results in impaired functionality, leading to maladaptive pain-related fear and anxiety and escape and avoidance behaviors. Neuroimaging studies in chronic pain patients have established that brain activity, especially in cortical and mesolimbic regions, is different from activity observed during acute pain in control subjects. In this review, we discuss the psychophysiological and neuronal factors that may be associated with the transition to chronic pain. We review information from human studies on neural circuits involved in emotional and motivational pain processing and how these circuits are altered in chronic pain conditions. We then highlight findings from animal research that can increase our understanding of the molecular and cellular mechanisms underlying emotional-motivational pain processing in the brain. Finally, we discuss how translational approaches incorporating results from both human and animal investigations may aid in accelerating the discovery of therapies.


Cloning of the koi herpesvirus (KHV) gene encoding thymidine kinase and its use for a highly sensitive PCR based diagnosis.
Outbreaks with mass mortality among common carp Cyprinus carpio carpio and koi Cyprinus carpio koi have occurred worldwide since 1998. The herpes-like virus isolated from diseased fish is different from Herpesvirus cyprini and channel catfish virus and was accordingly designated koi herpesvirus (KHV). Diagnosis of KHV infection based on viral isolation and current PCR assays has a limited sensitivity and therefore new tools for the diagnosis of KHV infections are necessary. A robust and sensitive PCR assay based on a defined gene sequence of KHV was developed to improve the diagnosis of KHV infection. From a KHV genomic library, a hypothetical thymidine kinase gene (TK) was identified, subcloned and expressed as a recombinant protein. Preliminary characterization of the recombinant TK showed that it has a kinase activity using dTTP but not dCTP as a substrate. A PCR assay based on primers selected from the defined DNA sequence of the TK gene was developed and resulted in a 409 bp amplified fragment. The TK based PCR assay did not amplify the DNAs of other fish herpesviruses such as Herpesvirus cyprini (CHV) and the channel catfish virus (CCV). The TK based PCR assay was specific for the detection of KHV and was able to detect as little as 10 fentograms of KHV DNA corresponding to 30 virions. The TK based PCR was compared to previously described PCR assays and to viral culture in diseased fish and was shown to be the most sensitive method of diagnosis of KHV infection. The TK based PCR assay developed in this work was shown to be specific for the detection of KHV. The TK based PCR assay was more sensitive for the detection of KHV than previously described PCR assays; it was as sensitive as virus isolation which is the golden standard method for KHV diagnosis and was able to detect as little as 10 fentograms of KHV DNA corresponding to 30 virions.


Research organizations in the Department of Veterans Affairs that focus on database analysis.
The Department of Veterans Affairs (VA) established several internal research organizations to analyze the many VA and non-VA databases. The information obtained assists VA in achieving its objective of providing cost-effective, high quality health care for veterans. Each group has a unique and well-defined function. All major informational inputs needed for VA planning are covered by the groups: resource allocation, workload requirements, population projections, and technology assessment, among others. Both managerial and academic research are conducted. The Department of Veterans Affairs has demonstrated flexibility in initiating new research center as the need arises, such as during the recent health care reform debates. The Department of Veterans Affairs has provided for the dissemination of information about research and databases by explicitly including this task as the focus of one group. A drawback to VA's research system is the minimal intercommunication among the groups. Learning about VA's internal research structure can assist other multihospital systems to identify their informational needs and establish suitable research organizations.


Polydimethylsiloxane microstructure-induced acoustic streaming for enhanced ultrasonic DNA fragmentation on a microfluidic chip.
Next-generation sequencing (NGS) is an essential technology for DNA identification in genomic research. DNA fragmentation is a critical step for NGS and doing this on-chip is of great interest for future integrated genomic solutions. Here we demonstrate fast acoustofluidic DNA fragmentation via ultrasound-actuated elastic polydimethylsiloxane (PDMS) microstructures that induce acoustic streaming and associated shear forces when placed in the field of an ultrasonic transducer. Indeed, acoustic streaming locally generates high tensile stresses that can mechanically stretch and break DNA molecule chains. The improvement in efficiency of the on-chip DNA fragmentation is due to the synergistic effect of these tensile stresses and ultrasonic cavitation phenomena. We tested these microstructure-induced effects in a DNA-containing microfluidic channel both experimentally and by simulation. The DNA fragmentation process was evaluated by measuring the change in the DNA fragment size over time. The chip works well with both long and short DNA chains; in particular, purified lambda (λ) DNA was cut from 48.5 kbp to 3 kbp in one minute with selected microstructures and further down to 300 bp within two and a half minutes. The fragment size of mouse genomic DNA was reduced from 1.4 kbp to 400 bp in one minute and then to 200 bp in two and a half minutes. The DNA fragmentation efficiency of the chip equipped with the PDMS microstructures was twice that of the chip without the microstructures. Exhaustive comparison shows that the on-chip fragmentation performance reaches the level of high-end professional standards. Recently, DNA fragmentation was shown to be enhanced using vibrating air microbubbles when the chip was placed in an acoustic field. We think the microbubble-free microstructure-based device we present is easier to operate and more reliable, as it avoids microbubble preparation and maintenance, while showing high DNA fragmentation performance.


Cell-mediated immunity predicts the probability of local recruitment in nestling blue tits.
We investigated whether the variation in T-cell-mediated immune function of blue tit nestlings affected their fledgling success and the probability of local survival. We studied the relationship between immune function and survival under two rearing conditions: control, unmanipulated, and experimentally enlarged broods. Brood enlargement had negative effects on nestling immune response. Immune response was positively related to fledgling success and it predicted the probability of local recruitment. However, the relationship between immune response and the probability of recruitment was significantly positive only among control broods and nonsignificant among enlarged broods. The effect of immune response on the recruitment probability was not affected by variation in body mass. Our study suggests that selection for immune responsiveness seems to be weak or even absent under unfavourable rearing conditions as simulated by brood size enlargement. Therefore, year-to-year environmental variation and environmental heterogeneity may constrain evolution towards higher immune responsiveness.


No Impact of Heat Stress and Dehydration on Short Duration Simulated Motor-Racing Performance.
Motor-racing drivers are often exposed to hot environments and may be susceptible to fluid loss and hydration issues, which could influence driving performance. This study assessed the effect of dehydration and heat stress on performance during a short, simulated motor-racing task. Nine healthy males (age: 26.6 ± 7.5 y, body mass: 78.8 ± 12.5 kg, mean ± SD) completed two passive dehydration (sauna) procedures (targeting -1% and -3% body mass loss (BML)) on separate occasions. Driving performance was assessed pre-dehydration (Baseline), immediately post-dehydration (Hot) and following a cooling period (Cool). Measures of driving performance included lap time and sector-time for one section of the track. Subjective ratings of mood, thermal stress and comfort were also collected during trials. Mean lap times were not different between Baseline, Hot, Cool conditions for both 1% (68.44 ± 1.43 s, 68.06 ± 1.17 s, 68.23 ± 1.25 s) and 3% (68.33 ± 1.68 s, 68.01 ± 1.15 s, 68.06 ± 1.26 s) trials respectively. In addition, mean sector times were not different between Baseline, Hot, Cool conditions for both 1% (11.61 ± 0.28 s, 11.55 ± 0.45 s, 11.59 ± 0.35 s) and 3% (11.49 ± 0.33 s, 11.56 ± 0.33 s, 11.63 ± 0.71 s) trials respectively. Changes in participants' subjective ratings (i.e. decreased alertness, concentration and comfort; increased tiredness and light-headedness) were observed at both levels of dehydration (1% and 3% BML), irrespective of heat stress. Thus, fluid loss and heat stress are unlikely to affect driver's motor-racing performance during short duration events. However, the impact of dehydration and heat stress on tasks of longer duration that accurately represent the demands associated with motor-racing requires further consideration.


Cross-Domain and Viral Interactions in the Microbiome.
The importance of the microbiome to human health is increasingly recognized and has become a major focus of recent research. However, much of the work has focused on a few aspects, particularly the bacterial component of the microbiome, most frequently in the gastrointestinal tract. Yet humans and other animals can be colonized by a wide array of organisms spanning all domains of life, including bacteria and archaea, unicellular eukaryotes such as fungi, multicellular eukaryotes such as helminths, and viruses. As they share the same host niches, they can compete with, synergize with, and antagonize each other, with potential impacts on their host. Here, we discuss these major groups making up the human microbiome, with a focus on how they interact with each other and their multicellular host.


siRNA knockdown of mitochondrial thymidine kinase 2 (TK2) sensitizes human tumor cells to gemcitabine.
Nucleoside metabolism enzymes are determinants of chemotherapeutic drug activity. The nucleoside salvage enzyme deoxycytidine kinase (dCK) activates gemcitabine (2', 2'-difluoro-2'-deoxycytidine) and is negatively regulated by deoxycytidine triphosphate (dCTP). Reduction of dCTP in tumor cells could, therefore, enhance gemcitabine activity. Mitochondrial thymidine kinase 2 (TK2) phosphorylates deoxycytidine to generate dCTP. We hypothesized that: (1) TK2 modulates human tumor cell sensitivity to gemcitabine, and (2) antisense knockdown of TK2 would decrease dCTP and increase dCK activity and gemcitabine activation. siRNA downregulation of TK2 sensitized MCF7 and HeLa cells (high and moderate TK2) but not A549 cells (low TK2) to gemcitabine. Combined treatment with TK2 siRNA and gemcitabine increased dCK. We also hypothesized that TK2 siRNA-induced drug sensitization results in mitochondrial damage that enhances gemcitabine effectiveness. TK2 siRNA and gemcitabine decreased mitochondrial redox status, DNA content, and activity. This is the first demonstration of a direct role for TK2 in gemcitabine resistance, or any independent role in cancer drug resistance, and further distinguishes TK2 function from that of other dTMP-producing enzymes [cytosolic TK1 and thymidylate synthase (TS)]. siRNA knockdown of TK1 and/or TS did not sensitize cancer cells to gemcitabine indicating that, among the 3 enzymes, only TK2 is a candidate therapeutic target for combination with gemcitabine.


Salvage intraosseous thrombolysis and extracorporeal membrane oxygenation for massive pulmonary embolism.
Intraosseous access is an alternative route of pharmacotherapy during cardiopulmonary resuscitation. Extracorporeal membrane oxygenation (ECMO) provides cardiac and respiratory support when conventional therapies fail. This case reports the use of intraosseous thrombolysis and ECMO in a patient with acute massive pulmonary embolism (PE). A 34-year-old female presented to the emergency department with sudden onset severe shortness of breath. Due to difficulty establishing intravenous access, an intraosseous needle was inserted into the left tibia. Echocardiography identified severe right ventricular dilatation with global systolic impairment and failure, indicative of PE. Due to the patient's hemodynamic compromise a recombinant tissue plasminogen activator (Alteplase) bolus was administered through the intraosseous route. After transfer to the intensive care unit, venous-arterial ECMO was initiated as further therapy. The patient recovered and was discharged 36 days after admission. This is the first report of combination intraosseous thrombolysis and ECMO as salvage therapy for massive PE.


Characterization and mutagenesis of two novel iron-sulphur cluster pentonate dehydratases.
We describe here the identification and characterization of two novel enzymes belonging to the IlvD/EDD protein family, the D-xylonate dehydratase from Caulobacter crescentus, Cc XyDHT, (EC 22.214.171.124), and the L-arabonate dehydratase from Rhizobium leguminosarum bv. trifolii, Rl ArDHT (EC 22.214.171.124), that produce the corresponding 2-keto-3-deoxy-sugar acids. There is only a very limited amount of characterization data available on pentonate dehydratases, even though the enzymes from these oxidative pathways have potential applications with plant biomass pentose sugars. The two bacterial enzymes share 41 % amino acid sequence identity and were expressed and purified from Escherichia coli as homotetrameric proteins. Both dehydratases were shown to accept pentonate and hexonate sugar acids as their substrates and require Mg(2+) for their activity. Cc XyDHT displayed the highest activity on D-xylonate and D-gluconate, while Rl ArDHT functioned best on D-fuconate, L-arabonate and D-galactonate. The configuration of the OH groups at C2 and C3 position of the sugar acid were shown to be critical, and the C4 configuration also contributed substantially to the substrate recognition. The two enzymes were also shown to contain an iron-sulphur [Fe-S] cluster. Our phylogenetic analysis and mutagenesis studies demonstrated that the three conserved cysteine residues in the aldonic acid dehydratase group of IlvD/EDD family members, those of C60, C128 and C201 in Cc XyDHT, and of C59, C127 and C200 in Rl ArDHT, are needed for coordination of the [Fe-S] cluster. The iron-sulphur cluster was shown to be crucial for the catalytic activity (kcat) but not for the substrate binding (Km) of the two pentonate dehydratases.