Selection of foot and mouth disease vaccine strains--a review.
The choice of the most appropriate strains of foot and mouth disease (FMD) virus vaccines to use in FMD control programmes and to store in vaccine antigen reserves is based on the matching of representative field isolates from outbreaks around the world to available vaccine strains. However, those involved in FMD control at a national level do not always give this work a high priority, while in countries without effective control of FMD there is little incentive to collect samples or to overcome the constraints on submission to international reference laboratories. In the short term, specific initiatives for targeted collection can provide samples on a periodic basis, but a long-term solution requires the development of FMD control measures. This must be underpinned by the strengthening of local Veterinary Services and laboratories, and by demand-driven provision of sufficient amounts of high-quality vaccine. Difficulties may be increased by commercial constraints on disclosure of the strains used for vaccine production and on the supply of reagents needed for matching tests. Vaccine matching tests are mainly based on in vitro methods - such as virus neutralisation, enzyme-linked immunosorbent assay with polyclonal antibodies and complement fixation - and are performed in a relatively small number of laboratories around the world. In addition to the difficulties of gathering representative field and vaccine strains, neither the reagents nor the methods used for vaccine matching are fully harmonised. Consequently, there is no strict equivalence in the results obtained. Alternative approaches using monoclonal antibody panels and/or viral capsid gene sequencing are being developed and could complement the currently employed serological tests. However, there is limited in vivo cross-protection information, more of which is essential for future validation of the vaccine matching methods. In response to the funding and leadership deficit for vaccine strain selection, a network of World Organisation for Animal Health (OIE) and Food and Agriculture Organization FMD reference laboratories has been established; this gives these laboratories the potential to strengthen the coordination of their work and reporting and thereby improve recommendations on vaccine strain selection.


Role of cytochrome P450 isoenzymes in metabolism of O(6)-benzylguanine: implications for dacarbazine activation.
O(6)-Benzylguanine (BG) effectively inactivates the DNA repair protein O(6)-alkylguanine-DNA alkyltransferase and enhances the effectiveness of alkylating agents, such as 1,3-bis(2-chloroethyl)-1-nitrosourea and temozolomide, in vitro and in vivo. BG is presently in clinical trials with 1,3-bis(2-chloroethyl)-1-nitrosourea and temozolomide. Preclinical data demonstrate that BG enhances the sensitivity of cells to 5-[3-methyl-triazen-1-yl]-imidazole-4-carboxamide, the active intermediate of dacarbazine (DTIC), making the combination BG plus DTIC attractive for additional clinical development. DTIC requires metabolism by cytochrome P450 (CYP450) isoforms, 1A1, 1A2, and 2E1 to form two reactive N-demethylated metabolites, 5-[3-hydroxy-methyl-3-methyl-triazen-1-yl]-imidazole-4-carboxamide and 5-[3-methyl-triazen-1-yl]-imidazole-4-carboxamide, ultimately forming a methylating species responsible for its cytotoxicity. The objective of this study was to examine the role of CYP450 1A1 and 1A2 in the metabolism of BG and identify possible drug-drug interactions with DTIC. Our data show that CYP450 isoforms 1A1 and 1A2 are primarily responsible for both BG oxidation to O(6)-benzyl-8-oxoguanine (8-oxoBG) and additional debenzylation to 8-oxoguanine. The catalytic efficiency of BG oxidation is 16 times lower for CYP1A1 than CYP1A2; however, the catalytic efficiency of 8-oxoBG debenzylation is 11 times greater for CYP1A1 than CYP1A2. Furthermore, BG inhibits CYP1A1 and 1A2 catalyzed conversion of DTIC to active methylating species. 8-OxoBG also inhibited conversion of DTIC to active methylating species but to a much lesser extent. The concentrations of BG required to inhibit 50% of DTIC N-demethylation were 2.8, 0.13, and 3.8 microM in human liver microsomes, baculovirus overexpressed CYP1A1, and CYP1A2, respectively. Our results suggest that treating patients with BG as a means to enhance the effectiveness of DTIC is unlikely to result in a therapeutic benefit as a result of inhibition of the enzymes responsible for conversion of DTIC to its active methylating species.


Enantioselective degradation and unidirectional chiral inversion of 2-phenylbutyric acid, an intermediate from linear alkylbenzene, by Xanthobacter flavus PA1.
Microbial degradation of the chiral 2-phenylbutyric acid (2-PBA), a metabolite of surfactant linear alkylbenzene sulfonates (LAS), was investigated using both racemic and enantiomer-pure compounds together with quantitative stereoselective analyses. A pure culture of bacteria, identified as Xanthobacter flavus strain PA1 isolated from the mangrove sediment of Hong Kong Mai Po Nature Reserve, was able to utilize the racemic 2-PBA as well as the single enantiomers as the sole source of carbon and energy. In the presence of the racemic compounds, X. flavus PA1 degraded both (R) and (S) forms of enantiomers to completion in a sequential manner in which the (S) enantiomer disappeared much faster than the (R) enantiomer. When the single pure enantiomer was supplied as the sole substrate, a unidirectional chiral inversion involving (S) enantiomer to (R) enantiomer was evident. No major difference was observed in the degradation intermediates with either of the individual enantiomers when used as the growth substrate. Two major degradation intermediates were detected and identified as 3-hydroxy-2-phenylbutanoic acid and 4-methyl-3-phenyloxetan-2-one, using a combination of liquid chromatography-mass spectrometry (LC-MS), and (1)H and (13)C nuclear magnetic resonance (NMR) spectroscopy. The biochemical degradation pathway follows an initial oxidation of the alkyl side chain before aromatic ring cleavage. This study reveals new evidence for enantiomeric inversion catalyzed by pure culture of environmental bacteria and emphasizes the significant differences between the two enantiomers in their environmental fates.


Spontaneous occurrence of hepatic lipidosis in a group of laboratory cats.
Clinical signs consistent with hepatic lipidosis occurred in six obese adult laboratory cats, housed in a group cage, 6 to 7 weeks after changing their diet from a commercial to a purified diet. The affected cats lost 30 to 40% of their body weight in this time period. This rate of weight loss is compatible with little or no food intake. For treatment, 5 cats were tube-fed three or four times daily with a high-fat liquid diet supplemented with L-citrulline and choline. All cats tolerated the diet, which contained 35% protein on an energy basis. Substantial voluntary food intake resumed 12 to 16 days after initiating treatment. The sixth cat was euthanatized. These observations suggest that 6 to 7 weeks of anorexia, associated with 30 to 40% weight loss, can induce hepatic lipidosis in obese but otherwise healthy cats, and confirm that with appropriate management the prognosis for cats with hepatic lipidosis is favorable.


Mach-Zehnder interferometry in a strongly driven superconducting qubit.
We demonstrate Mach-Zehnder-type interferometry in a superconducting flux qubit. The qubit is a tunable artificial atom, the ground and excited states of which exhibit an avoided crossing. Strongly driving the qubit with harmonic excitation sweeps it through the avoided crossing two times per period. Because the induced Landau-Zener transitions act as coherent beamsplitters, the accumulated phase between transitions, which varies with microwave amplitude, results in quantum interference fringes for n = 1 to 20 photon transitions. The generalization of optical Mach-Zehnder interferometry, performed in qubit phase space, provides an alternative means to manipulate and characterize the qubit in the strongly driven regime.


Occurrence of facultative anoxygenic photosynthesis among filamentous and unicellular cyanobacteria.
Eleven of 21 cyanobacteria strains examined are capable of facultative anoxygenic photosynthesis, as shown by their ability to photoassimilate CO2 in the presence of Na2S, 3-(3,4-dichlorophenyl)-1,1-dimethylurea and 703-nm light. These include different cyanobacterial types (filamentous and unicellular) of different growth histories (aerobic, anaerobic, and marine and freshwater). Oscillatoria limnetica, Aphanothece halophytica (7418), and Lyngbya (7104) have different optimal concentrations of Na2S permitting CO2 photoassimilation, above which the rate decreases: 3.5, 0.7, and 0.1 mM, respectively. In A. halophytica, for each CO2 molecule photoassimilated two sulfide molecules are oxidized to elemental sulfur, which is excreted from the cells. The ecological and evolutionary significance of anoxygenic photosynthesis in the cyanobacteria is discussed.


Treatment of elderly and other adult patients for depression in primary care.
To determine whether depression is treated differently in older and younger patients in primary care clinics. Administrative data were used to identify patients with a depression diagnosis code. The sources of data were baseline and 3-month follow-up surveys, the health plan electronic database, and chart audits. Nine primary care clinics owned by a health maintenance organization in the Midwest. The study sample (N=1023) consisted of adult patients, aged 19 to 93, and was divided into six age groups, from young adult, under age 35, to old old, 75 or older. Independent variables were a series of dummy variables: age groups, baseline depression severity, sex, and incident depression. Outcomes were defined as care processes (assessment, resources) and improvement in depression symptoms (Center for Epidemiologic Studies-Depression scale short form). Univariate and multivariate logistic regression analyses were used to analyze patient characteristics, depression symptoms, and care process variables. Significance level was reported based on the chi-square test of probability, P<or=.05. The adjusted response rates to the baseline and follow-up surveys were 69% and 82%. Elderly depressed patients were more likely than younger depressed patients to be widowed, have low levels of education, have fair or poor health, and have three or more comorbid health problems. There were no differences, by age, in number of depressive symptoms, antidepressant treatment, or recommendation for a follow-up appointment. Younger patients were more likely than older patients to have a new diagnosis of depression and to report being depressed most of the past year. Providers were only 6% as likely to ask old-old depressed patients about suicide risk, about one-fourth as likely to referral them to a mental health therapist, about one-fifth as likely to ask if they felt depressed, and one-twentieth as likely to ask about a problem with alcohol as they were with young-adult depressed patients. The old-old were about one-third as likely to report improvement in depression symptoms after 3 months as the young-adult patients. This study suggests a possible pattern of underattention to the oldest depressed patients. Failure to ask about suicide risk is especially of concern because of the high suicide rate of depressed geriatric patients. The finding that physicians are less likely to focus on depression with old-old patients may be explained, in part, by the concept of invisibility of the ordinary.


Estimates of quantal-release and binomial statistical-release parameters at rat neuromuscular junction.
Quantal-release and binomial statistical-release parameters were examined in the isolated rat diaphragm phrenic nerve preparation. The muscle resting potentials were reduced by cutting the muscle fibers to prevent muscle action potentials and contractions. The cutting technique was modified to allow persistent observation of miniature end-plate potentials (MEPPs) and end-plate potentials (EPPs) with intracellular recording techniques. Direct estimates of quantal release were examined and compared with predicted binomial and Poisson distributions. The results indicate that release is binomial when the nerve is stimulated with low- or high-frequency stimuli. The indirect method of estimating quantal release (variance method), which assumes release is described by a Poisson distribution, seriously overestimates quantal release. The binomial analysis indicates that the statistical store is small (less than 90 quanta) and that most of these quanta are released with each nerve impulse.


Clearance of apoptotic cells: implications in health and disease.
Recent advances in defining the molecular signaling pathways that regulate the phagocytosis of apoptotic cells have improved our understanding of this complex and evolutionarily conserved process. Studies in mice and humans suggest that the prompt removal of dying cells is crucial for immune tolerance and tissue homeostasis. Failed or defective clearance has emerged as an important contributing factor to a range of disease processes. This review addresses how specific molecular alterations of engulfment pathways are linked to pathogenic states. A better understanding of the apoptotic cell clearance process in healthy and diseased states could offer new therapeutic strategies.


Antiplatelet drugs and the perioperative period: What every urologist needs to know.
Antiplatelet agents like aspirin and clopidogrel are widely used for indications ranging from primary and secondary prevention of myocardial infarction or stroke to prevention of coronary stent thrombosis after percutaneous coronary interventions. When patients receiving antiplatelet drugs are scheduled for surgery, urologists commonly advise routine periprocedural withdrawal of these drugs to decrease the hemorrhagic risks that may be associated if such therapy is continued in the perioperative period. This approach may be inappropriate as stopping antiplatelet drugs often exposes the patient to a more serious risk, i.e. the risk of developing an arterial thrombosis with its potentially fatal consequences. Moreover, it has been seen that the increase in perioperative bleeding if such drugs are continued is usually of a quantitative nature and does not shift the bleeding complication to a higher risk quality. We, in this mini review, look at the physiological role and pathological implications of platelets, commonly used antiplatelet therapy and how continuation or discontinuation of such therapy in the perioperative period affects the hemorrhagic and thrombotic risks, respectively. Literature on the subject between 1985 and 2008 is reviewed. The consensus that seems to have emerged is that the policy of routine discontinuation of antiplatelet drugs in the perioperative period must be discouraged and risk stratification must be employed while making decisions regarding continuation or temporary discontinuation of antiplatelet therapy. Although antiplatelet drugs may be discontinued in patients at a low risk for an arterial thrombotic event, they must be continued in patients where the risks of bleeding and complications related to excessive bleeding are less than the risks of developing arterial thrombosis.


The influence of the O and K antigens of Klebsiella aerogenes on surface hydrophobicity and susceptibility to phagocytosis and antimicrobial agents.
The surface hydrophobicity of Klebsiella aerogenes is influenced by the presence of capsular (K) and lipopolysaccharide (O) antigens. Loss of both K and O antigens (K-O-), but not the K antigen alone (K-O+), increased surface hydrophobicity and susceptibility to phagocytosis. Unheated serum (i.e., containing complement) increased the surface hydrophobicity and phagocytosis of the K-O+ and K-O- strains, but not of the K+O+ encapsulated parent strain. Despite the altered susceptibility to phagocytosis caused by the presence or absence of the K and O antigens, their loss did not influence sensitivity to a range of hydrophilic, hydrophobic or cationic antimicrobial agents.


Serum Protein Biomarkers of Fibrosis Aid in Risk Stratification of Future Stricturing Complications in Pediatric Crohn's Disease.
Avoiding fibrostenotic complications is of paramount concern in the management of Crohn's disease (CD). We sought to investigate the association of candidate biomarkers of fibrosis collected at diagnosis with the future development of fibrostenotic CD. Using the Risk Stratification and Identification of Immunogenetic and Microbial Markers of Rapid Disease Progression in Children with Crohn's Disease cohort, a multicenter prospective observational pediatric inception cohort, subjects with an inflammatory phenotype (B1) at diagnosis who later converted to a stricturing phenotype (B2) within 3 years were compared with those who remained B1. Serum collected at diagnosis underwent both parallel reaction monitoring-targeted proteomic analysis and conventional enzyme-linked immunosorbent assay for 10 candidate biomarkers of intestinal fibrosis. Cox proportional hazard regression was used for multivariable analysis of time-dependent outcomes. In 116 subjects 58 subjects with verified B1 phenotype at diagnosis who later converted to B2 disease were compared with 58 subjects who remained B1 over 3 years of follow-up. Extracellular matrix protein 1 (ECM1) levels in the upper quartile (hazard ratio [HR] 3.43, 95% confidence limit [CL] 1.33, 8.42) were associated with future fibrostenotic disease. ASCA IgA (HR 4.99, 95% CL 1.50, 16.68) and CBir levels (HR 5.19, 95% CL 1.83, 14.74) were also associated with future intestinal fibrostenosis, although ECM1 continued to demonstrate independent association with conversion to B2 even with adjustment for serologies in multivariable analysis (HR 5.33, 95% CL 1.29, 22.13). ECM1 and other biomarkers of fibrosis may aid in determining the risk of uncomplicated inflammatory disease converting to B2 stricturing phenotypes in children with CD. Prospective validation studies to verify test performance and optimize clinical utilization are needed before clinical implementation.


Development of the pupillary light reflex from 9 to 24 months: association with common autism spectrum disorder (ASD) genetic liability and 3-year ASD diagnosis.
Although autism spectrum disorder (ASD) is heritable, the mechanisms through which genes contribute to symptom emergence remain unclear. Investigating candidate intermediate phenotypes such as the pupillary light reflex (PLR) prospectively from early in development could bridge genotype and behavioural phenotype. Using eye tracking, we longitudinally measured the PLR at 9, 14 and 24 months in a sample of infants (N = 264) enriched for a family history of ASD; 27 infants received an ASD diagnosis at 3 years. We examined the 9- to 24-month developmental trajectories of PLR constriction latency (onset; ms) and amplitude (%) and explored their relation to categorical 3-year ASD outcome, polygenic liability for ASD and dimensional 3-year social affect (SA) and repetitive/restrictive behaviour (RRB) traits. Polygenic scores for ASD (PGSASD ) were calculated for 190 infants. While infants showed a decrease in latency between 9 and 14 months, higher PGSASD was associated with a smaller decrease in latency in the first year (β = -.16, 95% CI = -0.31, -0.002); infants with later ASD showed a significantly steeper decrease in latency (a putative 'catch-up') between 14 and 24 months relative to those with other outcomes (typical: β = .54, 95% CI = 0.08, 0.99; other: β = .53, 95% CI = 0.02, 1.04). Latency development did not associate with later dimensional variation in ASD-related traits. In contrast, change in amplitude was not related to categorical ASD or genetics, but decreasing 9- to 14-month amplitude was associated with higher SA (β = .08, 95% CI = 0.01, 0.14) and RRB (β = .05, 95% CI = 0.004, 0.11) traits. These findings corroborate PLR development as possible intermediate phenotypes being linked to both genetic liability and phenotypic outcomes. Future work should incorporate alternative measures (e.g. functionally informed structural and genetic measures) to test whether distinct neural mechanisms underpin PLR alterations.


Pan-Cancer Analysis of lncRNA Regulation Supports Their Targeting of Cancer Genes in Each Tumor Context.
Long noncoding RNAs (lncRNAs) are commonly dysregulated in tumors, but only a handful are known to play pathophysiological roles in cancer. We inferred lncRNAs that dysregulate cancer pathways, oncogenes, and tumor suppressors (cancer genes) by modeling their effects on the activity of transcription factors, RNA-binding proteins, and microRNAs in 5,185 TCGA tumors and 1,019 ENCODE assays. Our predictions included hundreds of candidate onco- and tumor-suppressor lncRNAs (cancer lncRNAs) whose somatic alterations account for the dysregulation of dozens of cancer genes and pathways in each of 14 tumor contexts. To demonstrate proof of concept, we showed that perturbations targeting OIP5-AS1 (an inferred tumor suppressor) and TUG1 and WT1-AS (inferred onco-lncRNAs) dysregulated cancer genes and altered proliferation of breast and gynecologic cancer cells. Our analysis indicates that, although most lncRNAs are dysregulated in a tumor-specific manner, some, including OIP5-AS1, TUG1, NEAT1, MEG3, and TSIX, synergistically dysregulate cancer pathways in multiple tumor contexts.


A PRISMA-compliant meta-analysis of MDM4 genetic variants and cancer susceptibility.
Molecular epidemiological research suggests that mouse double minute 4 (MDM4) polymorphisms may be associated with cancer susceptibility, but results remain controversial. To derive a more precise evaluation, we performed a PRISMA compliant meta-analysis focused on five single nucleotide polymorphisms (rs11801299, rs1380576, rs10900598, rs1563828, and rs4245739) of MDM4. Overall, 23 studies involving 22,218 cases and 55,033 controls were analyzed. The results showed that rs4245739 was significantly associated with a decreased cancer risk in the allelic (C vs. A: odds ratio [OR] = 0.848, 95% confidence interval [CI] = 0.765-0.941, P = 0.002), heterozygous (AC vs. AA: OR = 0.831, 95% CI = 0.735-0.939, P = 0.003), and dominant (AC+CC vs. A: OR = 0.823, 95% CI = 0.727-0.932, P = 0.002) models. The association was more prominent in Asians. No significant association was found using any genetic model for the rs11801299, rs1380576, rs10900598, and rs1563828 SNPs. These results indicate that the rs4245739 polymorphism may contribute to a decreased cancer susceptibility and support the hypothesis that genetic variants in the MDM4 genes act as important modifiers of cancer risk.


Molecular epidemiology of Legionella pneumophila infection at a Canadian tertiary care institution.
To characterize the molecular epidemiology of Legionella species infection at one Canadian tertiary care centre. Twenty-eight clinical isolates and 12 environmental isolates obtained over a six-year period were analyzed by restriction fragment length polymorphism (rflp) of chromosomal dna. Isolates included 15 from 12 patients with hospital acquired illness and 13 from nine patients with community acquired infection. One nosocomial strain was Legionella micdadei and one community strain was Legionella pneumophila serotype 6; all others were L pneumophila serotype 1. rflp typing revealed one clone for all cases of a 1985 single-ward outbreak and five of six nonoutbreak L pneumophila nosocomial cases. An rflp pattern identical or highly related to that of the nosocomial clonal type was noted among nine of 12 L pneumophila serotype 1 community isolates. The remaining three isolates had two related rflp patterns distinct from the institutional strain. The nosocomial and community strains were isolated from multiple institutional water samples in the institution. For the environmental isolates, monoclonal antibody typing was more discriminating than rflp typing: seven monoclonal antibody subtypes were distinguished among 12 environmental isolates comprising three distinct rflp patterns. Despite multiple L pneumophila serotype 1 strains isolated in the authors' institutional water, a single clone of L pneumophila produced most disease. Community acquired disease was caused by a wider variety of strains.


Outcomes of cartilage repair techniques for chondral injury in the hip-a systematic review.
The aim of the study was to assess the options of treatment and their related outcomes for chondral injuries in the hip based on the available evidence whilst highlighting new and innovative techniques. A systematic review of the literature from PubMed (Medline), EMBASE, Google Scholar, British Nursing Index (BNI), Cumulative Index to Nursing and Allied Health Literature (CINAHL) and Allied and Complementary Medicine Database (AMED) was undertaken from their inception to March 2017 using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Clinical outcome studies, prospective/retrospective case series and case reports that described the outcome of cartilage repair technique for the chondral injury in the hip were included. Studies on total hip replacement, animal studies, basic studies, trial protocols and review articles were excluded. The systematic review found 21 relevant papers with 596 hips. Over 80% of the included studies were published in or after 2010. Most studies were case series or case reports (18 studies, 85.7%). Arthroscopy was used in 11 studies (52.4%). The minimum follow-up period was six months. Mean age of the participants was 37.2 years; 93.5% of patients had cartilage injuries of the acetabulum and 6.5% of them had injuries of the femoral head. Amongst the 11 techniques described in the systematic review, autologous matrix-induced chondrogenesis, osteochondral autograft transplantation and microfracture were the three frequently reported techniques. Over ten different techniques are available for cartilage repair in the hip, and most of them have good short- to medium-term outcomes. However, there are no robust comparative studies to assess superiority of one technique over another, and further research is required in this arena.


Bystander behavior in bullying situations: basic moral sensitivity, moral disengagement and defender self-efficacy.
The aim of the present study was to investigate how basic moral sensitivity in bullying, moral disengagement in bullying and defender self-efficacy were related to different bystander behaviors in bullying. Therefore, we examined pathways that linked students' basic moral sensitivity, moral disengagement, and defender self-efficacy to different bystander behaviors in bullying situations. Three hundred and forty-seven teenagers completed a bullying survey. Findings indicated that compared with boys, girls expressed higher basic moral sensitivity in bullying, lower defender self-efficacy and moral disengagement in bullying. Results from the SEM showed that basic moral sensitivity in bullying was negatively related to pro-bully behavior and positively related to outsider and defender behavior, mediated by moral disengagement in bullying, which in turn was positively related to pro-bully behavior and negatively related to outsider and defender behavior. What differed in the relations between outsider and defender behaviors was the degree of defender self-efficacy.


Roles of the amino terminal region and repeat region of the Plasmodium berghei circumsporozoite protein in parasite infectivity.
The circumsporozoite protein (CSP) plays a key role in malaria sporozoite infection of both mosquito salivary glands and the vertebrate host. The conserved Regions I and II have been well studied but little is known about the immunogenic central repeat region and the N-terminal region of the protein. Rodent malaria Plasmodium berghei parasites, in which the endogenous CS gene has been replaced with the avian Plasmodium gallinaceum CS (PgCS) sequence, develop normally in the A. stephensi mosquito midgut but the sporozoites are not infectious. We therefore generated P. berghei transgenic parasites carrying the PgCS gene, in which the repeat region was replaced with the homologous region of P. berghei CS (PbCS). A further line, in which both the N-terminal region and repeat region were replaced with the homologous regions of PbCS, was also generated. Introduction of the PbCS repeat region alone, into the PgCS gene, did not rescue sporozoite species-specific infectivity. However, the introduction of both the PbCS repeat region and the N-terminal region into the PgCS gene completely rescued infectivity, in both the mosquito vector and the mammalian host. Immunofluorescence experiments and western blot analysis revealed correct localization and proteolytic processing of CSP in the chimeric parasites. The results demonstrate, in vivo, that the repeat region of P. berghei CSP, alone, is unable to mediate sporozoite infectivity in either the mosquito or the mammalian host, but suggest an important role for the N-terminal region in sporozoite host cell invasion.


Integrated DNA methylation analysis identifies topographical and tumoral biomarkers in pilocytic astrocytomas.
Pilocytic astrocytoma (PA) is the most common glioma in pediatric patients and occurs in different locations. Chromosomal alterations are mostly located at chromosome 7q34 comprising the BRAF oncogene with consequent activation of the mitogen-activated protein kinase pathway. Although genetic and epigenetic alterations characterizing PA from different localizations have been reported, the role of epigenetic alterations in PA development is still not clear. The aim of this study was to investigate whether distinctive methylation patterns may define biologically relevant groups of PAs. Integrated DNA methylation analysis was performed on 20 PAs and 4 normal brain samples by Illumina Infinium HumanMethylation27 BeadChips. We identified distinct methylation profiles characterizing PAs from different locations (infratentorial vs supratentorial) and tumors with onset before and after 3 years of age. These results suggest that PA may be related to the specific brain site where the tumor arises from region-specific cells of origin. We identified and validated in silico the methylation alterations of some CpG islands. Furthermore, we evaluated the expression levels of selected differentially methylated genes and identified two biomarkers, one, IRX2, related to the tumor localization and the other, TOX2, as tumoral biomarker.


Metabolites Associated with Vigor to Frailty Among Community-Dwelling Older Black Men.
Black versus white older Americans are more likely to experience frailty, a condition associated with adverse health outcomes. To reduce racial disparities in health, a complete understanding of the pathophysiology of frailty is needed. Metabolomics may further our understanding by characterizing differences in the body during a vigorous versus frail state. We sought to identify metabolites and biological pathways associated with vigor to frailty among 287 black men ages 70-81 from the Health, Aging, and Body Composition study. Using liquid chromatography-mass spectrometry, 350 metabolites were measured in overnight-fasting plasma. The Scale of Aging Vigor in Epidemiology (SAVE) measured vigor to frailty based on weight change, strength, energy, gait speed, and physical activity. Thirty-seven metabolites correlated with SAVE scores (p < 0.05), while adjusting for age and site. Fourteen metabolites remained significant after multiple comparisons adjustment (false discovery rate < 0.30). Lower values of tryptophan, methionine, tyrosine, asparagine, C14:0 sphingomyelin, and 1-methylnicotinamide, and higher values of glucoronate, N-carbamoyl-beta-alanine, isocitrate, creatinine, C4-OH carnitine, cystathionine, hydroxyphenylacetate, and putrescine were associated with frailer SAVE scores. Pathway analyses identified nitrogen metabolism, aminoacyl-tRNA biosynthesis, and the citric acid cycle. Future studies need to confirm these SAVE-associated metabolites and pathways that may indicate novel mechanisms involved in the frailty syndrome.


NRT1.1-Related NH4 + Toxicity Is Associated with a Disturbed Balance between NH4 + Uptake and Assimilation.
A high concentration of ammonium (NH4 +) as the sole source of nitrogen in the growth medium often is toxic to plants. The nitrate transporter NRT1.1 is involved in mediating the effects of NH4 + toxicity; however, the mechanism remains undefined. In this study, wild-type Arabidopsis (Arabidopsis thaliana Columbia-0 [Col-0]) and NRT1.1 mutants (chl1-1 and chl1-5) were grown hydroponically in NH4NO3 and (NH4)2SO4 media to assess the function of NRT1.1 in NH4 + stress responses. All the plants grew normally in medium containing mixed nitrogen sources, but Col-0 displayed more chlorosis and lower biomass and photosynthesis than the NRT1.1 mutants in (NH4)2SO4 medium. Grafting experiments between Col-0 and chl1-5 further confirmed that NH4 + toxicity is influenced by NRT1.1. In (NH4)2SO4 medium, NRT1.1 induced the expression of NH4 + transporters, increasing NH4 + uptake. Additionally, the activities of glutamine synthetase and glutamate synthetase in roots of Col-0 plants decreased and soluble sugar accumulated significantly, whereas pyruvate kinase-mediated glycolysis was not affected, all of which contributed to NH4 + accumulation. By contrast, the NRT1.1 mutants showed reduced NH4 + accumulation and enhanced NH4 + assimilation through glutamine synthetase, glutamate synthetase, and glutamate dehydrogenase. Moreover, the up-regulation of genes involved in ethylene synthesis and senescence in Col-0 plants treated with (NH4)2SO4 suggests that ethylene is involved in NH4 + toxicity responses. This study showed that NH4 + toxicity is related to a nitrate-independent signaling function of NRT1.1 in Arabidopsis, characterized by enhanced NH4 + accumulation and altered NH4 + metabolism, which stimulates ethylene synthesis, leading to plant senescence.


Meta-mining of copy number profiles of high-risk neuroblastoma tumors.
Neuroblastoma, a pediatric tumor of the sympathetic nervous system, is predominantly driven by copy number aberrations, which predict survival outcome in global neuroblastoma cohorts and in low-risk cases. For high-risk patients there is still a need for better prognostic biomarkers. Via an international collaboration, we collected copy number profiles of 556 high-risk neuroblastomas generated on different array platforms. This manuscript describes the composition of the dataset, the methods used to process the data, including segmentation and aberration calling, and data validation. t-SNE analysis shows that samples cluster according to MYCN status, and shows a difference between array platforms. 97.3% of samples are characterized by the presence of segmental aberrations, in regions frequently affected in neuroblastoma. Focal aberrations affect genes known to be involved in neuroblastoma, such as ALK and LIN28B. To conclude, we compiled a unique large copy number dataset of high-risk neuroblastoma tumors, available via R2 and a Shiny web application. The availability of patient survival data allows to further investigate the prognostic value of copy number aberrations.


Na+-Ca2+ exchange in bovine rod outer segments requires and transports K+.
Intact outer segments isolated from bovine retinas (bovine ROS) display a high activity of Na+-Ca2+ exchange, and Na+-Ca2+ exchange appears to be the only functional ion transporter present. Here we demonstrate for the first time that Na+-Ca2+ exchange requires and transports K+ from the following observations. 1) Na+-Ca2+ exchange in bovine ROS required the simultaneous presence of K+ and Ca2+ on one side of the membrane and the presence of Na+ on the other side. 2) Na+-stimulated Ca2+ release from bovine ROS was accompanied by an equally large release of K+. We used the electrogenic protonophore carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) as an added electrical shunt; in the intact rod cell, electrogenic Na+-Ca2+ exchange is shunted by K+ channels present in the rod inner segment. In the presence of FCCP, an inward Na+-Ca2+ exchange current was accompanied by an outward current of protons with a stoichiometry of 1 H+/Ca2+; in the absence of FCCP, no Na+-induced proton current was observed. Addition of FCCP did not uncouple Na+-induced K+ release from Na+-induced Ca2+ release. We conclude that Na+-Ca2+ exchange in bovine rod photoreceptors operates at an electrogenic stoichiometry of 4 Na+:(1 Ca2+ + 1 K+). In isolated ROS and in the absence of an external electrical shunt, Na+-Ca2+ exchange operated at an electroneutral stoichiometry of 3 Na+:(1 Ca2+ + 1 K+).


20 years of medical surveillance on exposure to allergenic and non-allergenic platinum compounds: the importance of chemical speciation.
Chloroplatinates are potent allergens but other soluble platinum compounds such as tetraammine platinum dichloride (TPC) do not provoke reactions in subjects who are sensitive to chloroplatinates. TPC has been used in the manufacture of autocatalysts for 20 years. This study analyses 20 year data on exposure to soluble platinum compounds and medical surveillance to confirm that TPC is not allergenic. Workers in three distinct operations were exposed to soluble platinum compounds as chloroplatinates, chloroplatinates with TPC, or to TPC alone. Results of personal air sampling for soluble platinum compounds were compared together with the results of medical surveillance. The levels of exposure to soluble platinum compounds in each operation were comparable but the incidence of allergy was significantly different. In a subgroup of workers consistently exposed to chemical processes in each operation, the cumulative chance of being sensitised after 5 years of exposure was estimated as 51% for chloroplatinate exposure, 33% for mixed exposure, and 0% for TPC alone. The differences in sensitisation rates could not be explained by age, sex, and atopy. Nor could they be explained by the increased frequency of smoking in the workers with chloroplatinate exposure, despite the markedly higher risk of sensitisation in smokers. The differences could only be explained by the chemical stability of TPC. This study shows that the soluble platinum compound TPC is not allergenic under normal industrial conditions. Characterisation of the chemical compound (speciation) is essential to prevent stringent exposure limits being imposed for all soluble compounds on a generic basis.


Complete and Partial Lecithin:Cholesterol Acyltransferase Deficiency Is Differentially Associated With Atherosclerosis.
Lecithin:cholesterol acyltransferase (LCAT) is the sole enzyme that esterifies cholesterol in plasma. Its role in the supposed protection from atherogenesis remains unclear because mutations in LCAT causing fish-eye disease (FED) or familial LCAT deficiency (FLD) have been reported to be associated with more or instead less carotid atherosclerosis, respectively. This discrepancy may be associated with the loss of cholesterol esterification on only apolipoprotein AI (FED) or on both apolipoprotein AI- and apolipoprotein B-containing lipoproteins (FLD), an aspect that has thus far not been investigated. Seventy-four heterozygotes for LCAT mutations recruited from Italy and the Netherlands were assigned to FLD (n=33) or FED (n=41) groups and compared with 280 control subjects. Subclinical atherosclerosis was assessed with carotid intima-media thickness. Compared with control subjects, total cholesterol was lower by 16% (-32.9 mg/dL) and 7% (-14.9 mg/dL) and high-density lipoprotein cholesterol was lower by 29% (-16.7 mg/dL) and 36% (-20.7 mg/dL) in the FLD and FED groups, respectively. Subjects with FLD displayed a significant 18% lower low-density lipoprotein cholesterol compared with subjects with FED (101.9±35.0 versus 123.6±47.4 mg/dL; P=0.047) and control subjects (122.6±35.0 mg/dL; P=0.003). Remarkably, all 3 intima-media thickness parameters were lower in subjects with FLD compared with FED and control subjects (accounting for age, sex, body mass index, smoking, hypertension, family history of cardiovascular disease, and plasma lipids). After additional correction for nationality and ultrasonographic methods, average and maximum intima-media thickness remained significantly lower when subjects with FLD were compared with those with FED (0.59 versus 0.73 mm, P=0.003; and 0.87 versus 1.24 mm, P<0.001, respectively). In contrast, the common carotid intima-media thickness (corrected for age, sex, body mass index, smoking, hypertension, family history of cardiovascular disease, and plasma lipids) was higher in subjects with FED compared with control subjects (0.69 versus 0.65 mm; P=0.05), but this significance was lost after adjustment for nationality and ultrasonographic machine. In this head-to-head comparison, FLD and FED mutations were shown to be associated with decreased and increased atherosclerosis, respectively. We propose that this discrepancy is related to the capacity of LCAT to generate cholesterol esters on apolipoprotein B-containing lipoproteins. Although this capacity is lost in FLD, it is unaffected in FED. These results are important when considering LCAT as a target to decrease atherosclerosis.


Correlated and coupled cognitive change in older adults with and without preclinical dementia.
Common factor aging theories state that correlations among cognitive age effects signify a single underlying causal process. The logic underlying this proposition was evaluated by examining correlated cognitive change in a sample of 391 initially nondemented older adults who were tested annually for up to 16 years. Between-person correlations among rates of change (range = .56-.61) were partly attributable to model misspecification and the aggregation of heterogeneous groups of individuals. Correlated within-person cognitive change was much stronger in the cases (.45-.51) than in the noncases (.07-.18). These results demonstrate that correlated change may either signify causal commonality or the cumulative effects of multiple age-related conditions that can affect multiple cognitive systems.


Microscopic colitis: Current status, present and future challenges: statements of the European Microscopic Colitis Group.
Microscopic colitis (MC) is an inflammatory bowel disease presenting with chronic, non-bloody watery diarrhoea and few or no endoscopic abnormalities. The histological examination reveals mainly two subtypes of MC, lymphocytic or collagenous colitis. Despite the fact that the incidence in MC has been rising over the last decades, research has been sparse and our knowledge about MC remains limited. Specialists in the field have initiated the European Microscopic Colitis Group (EMCG) with the primary goal to create awareness on MC. The EMCG is furthermore a forum with the intention to promote clinical and basic research. In this article statements and comments are given that all members of the EMCG have considered being of importance for a better understanding of MC. The paper focuses on the newest updates in epidemiology, symptoms and diagnostic criteria, pathophysiology and highlights some unsolved problems. Moreover, a new treatment algorithm is proposed on the basis of new evidence from well-designed, randomized control trials.


Multiple abnormalities of myocardial insulin signaling in a porcine model of diet-induced obesity.
Heightened cardiovascular risk among patients with systemic insulin resistance is not fully explained by the extent of atherosclerosis. It is unknown whether myocardial insulin resistance accompanies systemic insulin resistance and contributes to increased cardiovascular risk. This study utilized a porcine model of diet-induced obesity to determine if myocardial insulin resistance develops in parallel with systemic insulin resistance and investigated potential mechanisms for such changes. Micropigs (n = 16) were assigned to control (low fat, no added sugars) or intervention (25% wt/wt coconut oil and 20% high-fructose corn syrup) diet for 7 mo. Intervention diet resulted in obesity, hypertension, and dyslipidemia. Systemic insulin resistance was manifest by elevated fasting glucose and insulin, abnormal response to intravenous glucose tolerance testing, and blunted skeletal muscle phosphatidylinositol-3-kinase (PI 3-kinase) activation and protein kinase B (Akt) phosphorylation in response to insulin. In myocardium, insulin-stimulated glucose uptake, PI 3-kinase activation, and Akt phosphorylation were also blunted in the intervention diet group. These findings were explained by increased myocardial content of p85alpha (regulatory subunit of PI 3-kinase), diminished association of PI 3-kinase with insulin receptor substrate (IRS)-1 in response to insulin, and increased serine-307 phosphorylation of IRS-1. Thus, in a porcine model of diet-induced obesity that recapitulates many characteristics of insulin-resistant patients, myocardial insulin resistance develops along with systemic insulin resistance and is associated with multiple abnormalities of insulin signaling.


Cellos: High-throughput deconvolution of 3D organoid dynamics at cellular resolution for cancer pharmacology.
Three-dimensional (3D) culture models, such as organoids, are flexible systems to interrogate cellular growth and morphology, multicellular spatial architecture, and cell interactions in response to drug treatment. However, new computational methods to segment and analyze 3D models at cellular resolution with sufficiently high throughput are needed to realize these possibilities. Here we report Cellos (Cell and Organoid Segmentation), an accurate, high throughput image analysis pipeline for 3D organoid and nuclear segmentation analysis. Cellos segments organoids in 3D using classical algorithms and segments nuclei using a Stardist-3D convolutional neural network which we trained on a manually annotated dataset of 3,862 cells from 36 organoids confocally imaged at 5 μm z-resolution. To evaluate the capabilities of Cellos we then analyzed 74,450 organoids with 1.65 million cells, from multiple experiments on triple negative breast cancer organoids containing clonal mixtures with complex cisplatin sensitivities. Cellos was able to accurately distinguish ratios of distinct fluorescently labelled cell populations in organoids, with ≤3% deviation from the seeding ratios in each well and was effective for both fluorescently labelled nuclei and independent DAPI stained datasets. Cellos was able to recapitulate traditional luminescence-based drug response quantifications by analyzing 3D images, including parallel analysis of multiple cancer clones in the same well. Moreover, Cellos was able to identify organoid and nuclear morphology feature changes associated with treatment. Finally, Cellos enables 3D analysis of cell spatial relationships, which we used to detect ecological affinity between cancer cells beyond what arises from local cell division or organoid composition. Cellos provides powerful tools to perform high throughput analysis for pharmacological testing and biological investigation of organoids based on 3D imaging.


Experimental far-field imaging properties of a ~5-μm diameter spherical lens.
Microscale lenses are mostly used as near-sighted lenses. The far-field imaging properties of a microscale spherical lens, where the lens is spatially separated from the object, are experimentally studied. Our experimental results show that, for a blu-ray disc (an object) whose spacing is 300 nm, the lens can magnify the stripe patterns of the disc when the lens is spatially separated from the object. In the experimentally tested range (0-14 μm), all the magnified images are virtual images. When the distance is increased from 0 to 14 μm the magnification decreases from 1.47× to 1.20× and the field of view increases from 3.8 to 12.2 μm. The image magnification cannot be described by standard geometrical optics.


Effect of breed of cattle on transmission rate and innate resistance to infection with Babesia bovis and B bigemina transmitted by Boophilus microplus.
To assess the effect of breed of cattle on the transmission rates of and innate resistance to Babesia bovis and B bigemina parasites transmitted by Boophilus microplus ticks. Groups of 56 purebred B indicus and 52 B indicus cross B taurus (50%, F1 generation) steers were placed in a paddock seeded with and also naturally infested with B microplus which were the progeny of females ticks fed on B taurus cattle specifically infected with a virulent isolate of B bovis. The cattle were placed in the infested paddock 50 days after seeding had started. Cattle were inspected from horseback daily for 50 days. Clinically ill cattle were brought to yards and assessed by monitoring fever, depression of packed-cell volume, parasitaemia and severity of clinical signs. Any animals that met preset criteria were treated for babesiosis. Blood samples were collected from all cattle on day 28, 35 and 42 after exposure and antibodies to Babesia spp and packed cell volume measured. All steers, except for one crossbred, seroconverted to B bovis and B bigemina by day 35 and 75% of the crossbred steers showed a maximum depression in packed cell volume of more than 15% due to infection with Babesia spp compared with only 36% of the B indicus group. Ten of the 52 crossbreds and 1 of the 56 B indicus steers showed severe clinical signs. Two of the crossbreds required treatment of which one died 2 weeks after initial treatment. Pure-bred B indicus cattle have a high degree of resistance to babesiosis, but crossbred cattle are sufficiently susceptible to warrant the use of preventive measures such as vaccination. Transmission rates of B bovis and B bigemina to B indicus and crossbred cattle previously unexposed to B microplus were the same.


Metabolite extraction for high-throughput FTICR-MS-based metabolomics of grapevine leaves.
In metabolomics there is an ever-growing need for faster and more comprehensive analysis methods to cope with the increase of biological studies. Direct infusion Fourier-transform ion cyclotron-resonance mass spectrometry (DI-FTICR-MS) is used in non-targeted metabolomics to obtain high-resolution snapshots of the metabolic state of a system. In any metabolic profiling study, the establishment of an effective metabolite extraction protocol is paramount. We developed an improved metabolite extraction method, compatible with DI-FTICR-MS-based metabolomics, using grapevine leaves. This extraction protocol allowed the extraction of polar and non-polar compounds, covering all major classes found in plants and increasing metabolome coverage.


Eating disorders in Spanish female athletes.
The objective of the study was to determine the prevalence of eating disorders and risk factors for their development in female athletes. Two hundred and eighty-three elite sportswomen, competing in 20 different sports, were administered the EAT, the CETCA (the Eating Disorders Assessment Questionnaire, based on DSM-III-R diagnostic criteria), and two other inventories which evaluated 1) the possible influence on eating disorders of exposure of the body in public and 2) pressure from coaches regarding eating habits, weight, physical appearance and performance. More than 11% of subjects had scores above the cut-off point (>30) on the EAT questionnaire, a proportion similar to that found in a general female population in Spain. On the basis of the CETCA score, AN was putatively diagnosed in 2.5% of the sample, and BN in 20.1%. Though some of these cases may have been EDNOS (eating disorders not otherwise specified), the proportion of athletes suffering from some kind of eating disorder was five times higher than in the general population (22.6% vs. 4.1%). No differences were found between the sportswomen and the general population in terms of specific risk behaviours and attitudes, but a substantial subgroup of athletes presented two or more of these risk behaviours. Exposure of the body in public seems to be a risk factor for eating disorders in general, and pressure from coaches seems to be a risk factor for bulimia.


Weak bases and ionophores rapidly and reversibly raise the pH of endocytic vesicles in cultured mouse fibroblasts.
It has been shown that endocytic vesicles in BALB/c 3T3 cells have a pH of 5.0 (Tycko and Maxfield, Cell, 28:643-651). In this paper, a method for measuring the effect of various agents, including weak bases and ionophores, on the pH of endocytic vesicles is presented. The method is based on the increase in fluorescein fluorescence with 490-nm excitation as the pH is raised above 5.0. Intensities of cells were measured using a microscope spectrofluorometer after internalization of fluorescein-labeled alpha 2-macroglobulin by receptor-mediated endocytosis. The increase in endocytic vesicle pH was determined from the increase in fluorescence after addition of various concentrations of the test agents. The following agents increased endocytic vesicle pH above 6.0 at the indicated concentrations: monensin (6 microM), FCCP (10 microM), chloroquine (140 microM), ammonia (5 mM), methylamine (10 mM). The ability of many of these agents to raise endocytic vesicle pH may account for many of their effects on receptor-mediated endocytosis. Dansylcadaverine caused no effect on vesicle pH at 1 mM. The observed increases in vesicle pH were rapid (1-2 min) and could be reversed by removal of the perturbant. This reversibility indicates that the vesicles themselves contain a mechanism for acidification. The increase in vesicle pH due to these treatments can be observed visually using an SIT video camera. Using this method, it is shown that endocytic vesicles become acidic at very early times (i.e., within 5-7 min of continuous uptake at 37 degrees C).


Oral Candida in HIV infection and AIDS: new perspectives/new approaches.
Oral candidosis has become an increasingly important problem in HIV-infected individuals. At present, the small body of published literature on the characterization of the Candida strains and species found in HIV+ patients is full of confusion and contradictions. Some of these difficulties are the result of the methodological shortcomings of a number of the techniques that have been used. Examples of the problems that may be encountered on primary isolation and subculture are described and the drawbacks associated with the systems used to date for phenotyping Candida are quoted. While molecular characterization techniques would appear to offer a reliable and objective alternative, they too have their strengths and weaknesses. An attempt is made to summarize the progress that has been made recently in the detection and identification of Candida albicans and also the non-albicans species from HIV-infected individuals. What emerges is that the commensal Candida species that inhabit the oral cavities of HIV+ patients are subjected to a number of significant pressures that probably promote the selection of organisms with unusual phenotypes and genotypes. These Candida are more difficult to characterize and behave differently compared to their counterparts in HIV- individuals. It is clear that uncovering the factors that are important for the selection of treatment regimens and will be predictive of outcome will not be easy. Candida organisms are neither as benign nor as simple as once thought.


Phase III double-blind, randomized, placebo-controlled crossover trial of black cohosh in the management of hot flashes: NCCTG Trial N01CC1.
Hot flashes can cause significant morbidity in postmenopausal women undergoing or finished with breast cancer treatment. Black cohosh has been used to treat hot flashes, but definitive clinical data about efficacy have been equivocal. A double-blind, randomized, cross-over clinical trial with two 4-week periods, was used to study the efficacy of black cohosh (1 capsule, Cimicifuga racemosa 20 mg BID) for the treatment of hot flashes in women. Participants kept a daily hot flash diary during a baseline week and then during two 4-week crossover treatment periods. Hot flash scores were measured by assigning points (1 to 4 for mild to very severe) to each hot flash based on severity and then adding the points for a given time period. Between October 31, 2003, to March 4, 2004, 132 patients were randomly assigned. Toxicity was minimal and not different by treatment group. Patients receiving black cohosh reported a mean decrease in hot flash score of 20% (comparing the fourth treatment week to the baseline week) compared with a 27% decrease for patients on placebo (P = .53). Mean hot flash frequency was reduced 17% on black cohosh and 26% on placebo (P = .36). Patient treatment preferences were measured after completion of both treatment periods by ascertaining which treatment period, if any, the patient preferred. Thirty-four percent of patients preferred the black cohosh treatment, 38% preferred the placebo, and 28% did not prefer either treatment. This trial failed to provide any evidence that black cohosh reduced hot flashes more than the placebo.


Inequalities in mental health and well-being in a time of austerity: Baseline findings from the Stockton-on-Tees cohort study.
Since 2010, the UK has pursued a policy of austerity characterised by public spending cuts and welfare changes. There has been speculation - but little actual research - about the effects of this policy on health inequalities. This paper reports on a case study of local health inequalities in the local authority of Stockton-on-Tees in the North East of England, an area characterised by high spatial and socio-economic inequalities. The paper presents baseline findings from a prospective cohort study of inequalities in mental health and mental wellbeing between the most and least deprived areas of Stockton-on-Tees. This is the first quantitative study to explore local mental health inequalities during the current period of austerity and the first UK study to empirically examine the relative contributions of material, psychosocial and behavioural determinants in explaining the gap. Using a stratified random sampling technique, the data was analysed using multi-level models that explore the gap in mental health and wellbeing between people from the most and least deprived areas of the local authority, and the relative contributions of material, psychosocial and behavioural factors to this gap. The main findings indicate that there is a significant gap in mental health between the two areas, and that material and psychosocial factors appear to underpin this gap. The findings are discussed in relation to the context of the continuing programme of welfare changes and public spending cuts in the UK.


A comparison between stainless steel crowns and multisurface amalgams in primary molars.
From the findings in this study, it appears that steel crowns for primary teeth are significantly superior to multisurface amalgam fillings, in terms of the percentage of teeth requiring subsequent care: 88.7 percent of the teeth initially restored with amalgam required follow-up treatment, and only 30.3 percent of the crowned teeth required such treatment. Because both types of restorations were studied for approximately the same length of time, one could not justly conclude that the crowns were superior in terms of longevity. Steel crowns may prove to be more economical for the patient than amalgam fillings, if follow-up visits are considered. The author plans to conduct a second study on the cost factor.


[Advances in antibody therapy for B cell lymphoma].
Almost 20 years have passed after rituximab, an anti-CD20 mouse-human chimeric monoclonal antibody therapeutic, was introduced in the clinical setting for the treatment of CD20-positive B cell malignancies. Although the prognosis has significantly improved in most B cell malignancies, resistance to rituximab with/without chemotherapies is also widely recognized. With this background, newer generation antibody therapeutics and other molecular targeting drugs that show more effectiveness especially to refractory patients are critically required. In this review, molecular mechanisms of the second/third generation anti-CD20 antibodies (ofatumumab and obinutuzumab), anti-CD19 chimeric antigen-receptor T-cells (CAR-T-CD19; tisagenlecleucel, axicabtagene ciloleucel), and the anti-PD-1 antibodies (nivolumab and pembrolizumab) are presented. The therapeutic effectiveness of those drugs as monotherapy and/or combined therapy with conventional chemotherapy (immunochemotherapy) and another molecular targeting therapeutics (so-called "chemo-free" regimens) are also reviewed.


Can an intraductal papillary mucinous tumor be a potential indicator of concurrent adenocarcinoma of the pancreas?
Despite the recent progress of diagnostic and therapeutic modalities, survival rates of pancreatic adenocarcinoma remain poor, mainly due to late diagnosis. We report the case of a 56-year-old man who was diagnosed with a symptomatic intraductal papillary mucinous tumor of the pancreas located in the uncus. This tumor was associated with a concurrent stenosis of the isthmic pancreatic duct which resulted in a distal dilation. A Whipple procedure was performed. During the procedure, a concomitant adenocarcinoma was diagnosed 2 cm from the primary intraductal papillary mucinous tumor, causing the isthmic stenosis. A second resection was then performed to the left of the pancreatic isthmus, and adjuvant chemotherapy was performed. The patient is well and without any sign of recurrence 7 months after surgery. We discuss the possibility that intraductal papillary mucinous tumors may be a "red flag" enabling earlier diagnosis of a concurrent pancreatic adenocarcinoma arising in another area of the pancreas.


Transcriptomic analysis reveals novel age-independent immunomodulatory proteins as a mode of cerebroprotection in P2X4R KO mice after ischemic stroke.
Identification of new potential drug target proteins and their plausible mechanisms for stroke treatment is critically needed. We previously showed that genetic deletion and short-term pharmacological inhibition of P2X4R, a purinergic receptor for adenosine triphosphate ATP, provides acute cerebroprotection. However, potential mechanisms remain unknown. Therefore, we employed RNA-seq technology to identify the gene expression profiles, pathway analysis, and qPCR validation of differentially expressed genes (DEGs). This analysis identified roles of DEGs in certain biological processes responsible for P2X4R-dependent cerebroprotection after stroke. We subjected both young and aged male and female global P2X4 KO and littermate WT mice to ischemic stroke. After 3 days, mice were sacrificed, total RNA was isolated using Trizol, and subjected to RNA-seq and Nanostring-mediated qPCR. DESeq2, Gene Ontology (GO), and Ingenuity Pathway Analysis (IPA) were used to identify mRNA transcript expression profiles and biological pathways. We found 2246 DEGs in P2X4R KO vs WT tissue after stroke. Out of these DEGs, 1920 gene were downregulated, and 325 genes were upregulated in KO. GO/IPA analysis of the top 300 DEGs suggests an enrichment of inflammation and extracellular matrix component genes. qPCR validation of the top 30 DEGs revealed downregulation of two common age-independent genes in P2X4R KO mice: Interleukin-6 ( IL-6) , an inflammatory cytokine, and Cytotoxic T Lymphocyte-Associated Protein 2 alpha ( Ctla2a ), an immunosuppressive factor. These data suggest that P2X4R-mediated cerebroprotection after stroke is initiated by attenuation of immune modulatory pathways in both young and aged mice of both sexes.


Developments in understanding acquired immunity and innate susceptibility to norovirus and rotavirus gastroenteritis in children.
We discuss recent advances in the understanding of acquired immunity and susceptibility to the two major pediatric enteric viral pathogens, norovirus and rotavirus. The prominent decline in severe rotavirus gastroenteritis in areas with mature rotavirus vaccination programmes has correspondingly unmasked the significant burden of disease associated with norovirus gastroenteritis among children. As epidemiologists and vaccinologists set their sights on this next vaccine target, we provide an update on norovirus vaccine development.In addition to these developments regarding acquired immunity, refinements to our understanding of innate susceptibility to norovirus has advanced. Significant recent advances now describe similar immunologic mechanisms in understanding susceptibility for both norovirus and rotavirus, involving histo-blood group antigenic associations, which may also prove to be genotype specific. This information can potentially be used to tailor both applied and developmental efforts to public health interventions against these important pediatric enteric viral pathogens.


Fusion of the homeobox gene HLXB9 and the ETV6 gene in infant acute myeloid leukemias with the t(7;12)(q36;p13).
Recently, we and others reported a recurrent t(7;12)(q36;p13) found in myeloid malignancies in children < or =18 months of age and associated with a poor prognosis. Fluorescence in situ hybridization studies mapped the 12p13 breakpoint to the first intron of ETV6 and narrowed down the region of 7q36 involved. By using the sequences made public recently by the Human Genome Project, two candidate genes in 7q36 were identified: the homeobox gene HLXB9 and c7orf3, a gene with unknown function. Reverse transcription-PCR of two cases with t(7;12), using primers for c7orf3 and ETV6, was negative. However, reverse transcription-PCR for HLXB9-ETV6 demonstrated alternative splicing; the two major bands corresponded to fusion of exon 1 of HLXB9 to exons 2 and 3, respectively, of ETV6. The reciprocal ETV6-HLXB9 transcript was not detected. It remains to be elucidated if the leukemic phenotype is attributable to the formation of the HLXB9-ETV6 fusion protein, which includes the helix-loop-helix and E26 transformation-specific DNA binding domains of ETV6 or to the disruption of the normal ETV6 protein.


[Predictive value of psychiatric symptoms in the preschool age for adjustment in school age].
This paper derives from a prospective cohort study conducted in Freiburg (Federal Republic of Germany) from 1979 to 1987 with 209 (follow-up outcome: 172) 3- to 5-year-olds. The "Behaviour-Screening Questionnaire," the "Children's Global Assessment Scale" and a thorough semi-standardized interview were used to identify behavior problems in the children. The results were analyzed in relation to life events, sex and socioeconomic status. They were compared with the findings in other studies. Most of the variables examined proved to be of low predictive value. The mood changes common in preschool and kindergarten children with behavior problems were no longer in evidence 7 years later.


Evaluation and management of chronic kidney disease: synopsis of the kidney disease: improving global outcomes 2012 clinical practice guideline.
The Kidney Disease: Improving Global Outcomes (KDIGO) organization developed clinical practice guidelines in 2012 to provide guidance on the evaluation, management, and treatment of chronic kidney disease (CKD) in adults and children who are not receiving renal replacement therapy. The KDIGO CKD Guideline Development Work Group defined the scope of the guideline, gathered evidence, determined topics for systematic review, and graded the quality of evidence that had been summarized by an evidence review team. Searches of the English-language literature were conducted through November 2012. Final modification of the guidelines was informed by the KDIGO Board of Directors and a public review process involving registered stakeholders. The full guideline included 110 recommendations. This synopsis focuses on 10 key recommendations pertinent to definition, classification, monitoring, and management of CKD in adults.


Synergistic augmentation of rapamycin-induced autophagy in malignant glioma cells by phosphatidylinositol 3-kinase/protein kinase B inhibitors.
The mammalian target of rapamycin (mTOR) is a downstream effector of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway and a central modulator of cell proliferation in malignant gliomas. Therefore, the targeting of mTOR signaling is considered a promising therapy for malignant gliomas. However, the mechanisms underlying the cytotoxic effects of a selective mTOR inhibitor, rapamycin, on malignant glioma cells are poorly understood. The purpose of this study was thus to elucidate how rapamycin exerts its cytotoxic effects on malignant glioma cells. We showed that rapamycin induced autophagy but not apoptosis in rapamycin-sensitive malignant glioma U87-MG and T98G cells by inhibiting the function of mTOR. In contrast, in rapamycin-resistant U373-MG cells, the inhibitory effect of rapamycin was minor, although the phosphorylation of p70S6 kinase, a molecule downstream of mTOR, was remarkably inhibited. Interestingly, a PI3K inhibitor, LY294002, and an Akt inhibitor, UCN-01 (7-hydroxystaurosporine), both synergistically sensitized U87-MG and T98G cells as well as U373-MG cells to rapamycin by stimulating the induction of autophagy. Enforced expression of active Akt in tumor cells suppressed the combined effects of LY294002 or UCN-01, whereas dominant-negative Akt expression was sufficient to increase the sensitivity of tumor cells to rapamycin. These results indicate that rapamycin exerts its antitumor effect on malignant glioma cells by inducing autophagy and suggest that in malignant glioma cells a disruption of the PI3K/Akt signaling pathway could greatly enhance the effectiveness of mTOR inhibitors.


Co-production of 155Tb and 152Tb irradiating 155Gd / 151Eu tandem target with a medium energy α-particle beam.
Four terbium isotopes 149,152,155,161Tb emitting various types of radiation can be used for both diagnostics and therapy. 152Tb emits positrons and is ideal for PET. 155Tb is considered a promising Auger emitter and a diagnostic pair for other terbium therapeutic isotopes. Several methods for the production of 155Tb using charged particle accelerators have been proposed, but they all have significant limitations. The restricted availability of this isotope hinders its medical applications. We have proposed a new method for production of 155Tb, irradiating enriched 155Gd by alpha particles. The possibility of simultaneous production of two isotopes of terbium, 152,155Tb, was also studied for more efficient cyclotron beam use. Irradiation of 155Gd enriched targets and 155Gd / 151Eu tandem target with alpha-particles with an energy of 54 MeV was carried out at the U-150 cyclotron at the NRC "Kurchatov Institute". The cross sections of nuclear reactions on enr-155Gd were measured by the stack foil technique, detecting the gamma-radiation of the activation products. The separation of rare earth elements was performed by extraction chromatography with the LN Resin. 155Tb was produced via 155Dy decay. The cross sections for the 155,156Tb and 155,157Dy production were measured by the irradiation of a gadolinium target enriched with the 155Gd isotope with alpha-particles in an energy range of 54 → 33 MeV. The yield of 155Dy on a thick target at 54 MeV was 130 MBq/μAh, which makes it possible to obtain 1 GBq of 155Tb in 11 hour-irradiation with 20 μA beam current. The possibility of simultaneous production of 152,155Tb by irradiation of 155Gd and 151Eu tandem target with medium-energy alpha-particles is implemented. Optimal irradiation energy ranges of alpha -particles as 54 → 42 MeV for 155Tb and 42 → 34 MeV for 152Tb were suggested. Product activity and radionuclidic purity were calculated.


Studies on retinal and retinal pigment epithelial gene expression.
The 'completion' of the murine and human genomes and creation of high-density expressed sequence tag (EST) databases from multiple tissues and multiple species, coupled with the development of high-throughput expression profiling approaches such as microarrays and Serial Analysis of Gene Expression (SAGE), is making possible the in-depth analysis of gene expression patterns in health and disease to an extent that was not previously possible. Such new information is providing insight into normal function, and into how normal function is altered in disease. Efforts have begun, and are accelerating, in the application of expression profiling to the study of the retina and retinal pigment epithelium (RPE). In this chapter we will review progress in this area. We will also discuss technical issues that make expression studies of the RPE particularly challenging, and share our experience in methodological approaches to overcome these challenges.


Optimum on-axis computer-generated hologram encoded into low-resolution phase-modulation devices.
A computer-generated hologram with minimum spatial complexity is proposed for encoding arbitrary complex transmittance with low-resolution phase-modulation devices. The phase modulation of the proposed hologram is determined such that on-axis reconstruction with high signal-to-noise ratio, optimum reconstruction efficiency, and the largest possible signal bandwidth can be obtained.


Comparison of radioimmunoassay with thin-layer chromatographic and gas-liquid chromatographic methods of barbiturate detection in human urine.
A radioimmunoassay (I) for barbiturates was compared with thin-layer chromatographic (II) and gas-liquid chromatographic (III) methods for barbiturate detection in human urine. Timed urine samples were obtained from volunteers who had ingested 100 mg of a barbiturate. I detected barbiturate in all urines tested up to 76 h after the dose, and III in all up to 52 h and in 90% up to 76 h. II detected barbiturates in 90% of all urine samples for only 30 h, after which is reliability declined. Glutethimide interfered with radioimmunoassay of barbiturate, producing false positives. I is sensitive, reliable, and fast, and lends itself to screening large numbers of urine samples for barbiturates. For routine urine surveillance, however, we found I to be less useful than II, which is still the method of choice. I has, however, proved to be an excellent method for confirming results of II.


Cancer incidence in a population with a high prevalence of infection with human immunodeficiency virus type 1.
Human immunodeficiency virus type 1 (HIV-1) infection is known to increase the incidence of Kaposi's sarcoma and non-Hodgkin's lymphoma. Parallels with other causes of immunodeficiency suggest a possible effect of HIV-1 on additional cancers. This study was designed to determine the types and rates of cancers occurring in excess in the presence of HIV-1 infection. We examined cancer incidence in a population-based open cohort with a high prevalence of HIV-1 infection. The study population was never-married men aged 25-54 years who resided in San Francisco, Calif., of whom an estimated 20,000 (24%) were HIV-1 seropositive as of late 1984. Cancer registration data covering 1,390,000 person-years of observation of these men from 1973 through 1990 were obtained from the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) Program. Standardized incidence rates and ratios of observed to expected cases (based on rates in the pre-acquired immunodeficiency syndrome [pre-AIDS] period [i.e., 1973-1979]) were calculated for cancers classified by site and by cell type. The incidence of Kaposi's sarcoma in never-married men plateaued in 1988-1990 at 0.5% per year. The incidence of non-Hodgkin's lymphoma increased 20-fold between 1973-1979 and 1988-1990; increases were most pronounced in tumors of higher grade histology and extranodal (especially central nervous system) primary sites. The incidence of Burkitt's and Burkitt-like tumors peaked in 1985-1987, whereas that of large cell diffuse and immunoblastic lymphomas increased throughout the study period. The incidence of Hodgkin's disease was 2.0 (95% confidence interval [CI] = 1.3-3.0) times expected in 1988-1990. The incidence of anal cancer was 9.9 (95% CI = 4.5-18.7) times expected in 1973-1979 and 10.1 (95% CI = 5.0-18.0) times expected in 1988-1990. Ratios of observed to expected cancers of most other sites were 2.0 or less; the ratio of leiomyosarcomas (at any site) was 2.5 (95% CI = 0.5-7.4). As the HIV-1 epidemic has progressed, the increases in AIDS-related Kaposi's sarcoma, Burkitt's tumor, and other non-Hodgkin's lymphoma have followed different patterns. The effect of HIV-1 on other cancers has been nondetectable. In particular, HIV-1 is not related to the increased risk of anal cancer in homosexual men, which antedated the AIDS epidemic. These data suggest that the etiologic mechanisms of HIV-1-related malignancy differ for specific cancers and do not globally increase cancer risk. Control of HIV-1-related cancer remains an unsolved challenge in the management of HIV-1 infection.


Encapsulation of magnetic and fluorescent nanoparticles in emulsion droplets.
Oils containing both fluorescent semiconductor and magnetic oxide nanoparticles are used to produce oil in water emulsions. This technique produces oil droplets with homogeneous fluorescence and high magnetic nanoparticle concentrations. The optical properties of the oil droplets are studied as a function of the droplet sizes for various concentrations of fluorescent and magnetic nanoparticles. For all concentrations tested, we find a linear variation of the droplet fluorescent intensity as a function of the droplet volume. For a given size and a given quantum dot (QD) concentration, the droplet fluorescence intensity drops sharply as a function of the magnetic nanoparticle concentration. We show that this decrease is due mainly to the strong absorption cross section of the magnetic nanoparticles and to a lesser extent to the dynamic and static quenching of the QD fluorescence. The role of the iron oxide nanoparticle localization in the droplet (surface versus volume) is also discussed.


Metabolic activity of liver mitochondria from magnesium-deficient rats.
Comparisons were made between the metabolic activities of whole mitochondria and intact mitochondrial inner membrane preparations from magnesium-deficient and control rats in a basic medium without exogenous magnesium. Magnesium deficiency partially uncoupled oxidative phosphorylation in whole mitochondria and completely uncoupled it with inner membrane preparations. Addition of 1 mM MgCl2 to the medium prevented the total uncoupling of inner membranes and gave ADP:O values similar to those obtained with whole mitochondria from magnesium-deficient rats. No impairment in proton extrusion by intact inner membranes or in ATPase activity by either intact or fragmented inner membranes was detected during magnesium deficiency, but there was evidence of increased membrane permeability to the inward movement of protons. It is concluded that magnesium deficiency probably increases the permeability of the mitochondrial inner membrane and this weakens the coupling between oxidation and phosphorylation.


Radiolabeled oligonucleotides for antisense imaging.
Oligonucleotides radiolabeled with isotopes emitting γ-rays (for SPECT imaging) or positrons (for PET imaging) can be useful for targeting messenger RNA (mRNA) thereby serving as non-invasive imaging tools for detection of gene expression in vivo (antisense imaging). Radiolabeled oligonucleotides may also be used for monitoring their in vivo fate, thereby helping us better understand the barriers to its delivery for antisense targeting. These developments have led to a new area of molecular imaging and targeting, utilizing radiolabeled antisense oligonucleotides. However, the success of antisense imaging relies heavily on overcoming the barriers for its targeted delivery in vivo. Furthermore, the low ability of the radiolabeled antisense oligonucleotide to subsequently internalize into the cell and hybridize with its target mRNA poses additional challenges in realizing its potentials. This review covers the advances in the antisense imaging probe development for PET and SPECT, with an emphasis on radiolabeling strategies, stability, delivery and in vivo targeting.


[Disturbances in ammonia metabolism in hepatic failure].
Serious hepatocellular dysfunction leads to disturbances in the ammonia metabolism and increases the risk for development of hepatic encephalopathy. The present paper describes pathophysiological patterns for development of hyperammonemia in patients with liver diseases and discusses the rationale for novel ammonia lowering strategies. The present paper is based on experimental research performed in a porcine model of acute liver failure and a literature search in Pubmed. Under normal conditions the intestines and the kidneys are the main ammonia producing organs, whereas the liver and skeletal muscles are the major ammonia consuming organs. Small and large intestines contribute equally to the release of ammonia. Development of intra- and extra hepatic portasystemic shunts, impaired urea synthesis, and reduced hepatic perivenous glutamine synthesis capacity, contribute to the development of hyperammonaemia with hepatic failure. The kidneys are quantitatively as important as the portal drained viscera in ammonia production, but can adapt to hyperammonaemia by reducing the production of ammonia to the blood and at the same time increase its excretion in urine. Furthermore, the lungs are metabolically active, but their role in hyperammonaemia remains unsettled. Skeletal muscle contributes to reducing the ammonia level in the body through synthesis of glutamine from equimolar quantities of glutamate and ammonia. Current therapeutic recommendations on protein restriction, use of antibiotics and lactulosis to patients with hepatic failure are not sufficiently documented. There is a need for development of novel ammonia lowering strategies.


A retrospective analysis of vinorelbine chemotherapy for patients with previously treated soft-tissue sarcomas.
Introduction. The role of vinorelbine in specific soft tissue sarcoma subtypes is unclear. We present retrospective single institution experience with single-agent vinorelbine in subjects with metastatic soft tissue malignancies. Methods. Fifty-eight patients were treated with single agent intravenous vinorelbine between April 1997 and December 2004. Doxorubicin had been administered previously to 53 subjects (91%), and the median number of lines of previous chemotherapy was 3 (range 0-7). Results. Patients received a median 6 doses of vinorelbine (range 1-65). The overall response rate was 6% (3 patients: 1 angiosarcoma, 1 epithelioid sarcoma, and 1 embryonal rhabdomyosarcoma). Fourteen patients (26%) experienced a best result of stable disease. Median time to progression was 1.8 months (95% confidence intervals 1.5-2.1 months, Kaplan-Meier estimate). Eight patients experienced grade 3 or 4 toxicity, most commonly febrile neutropenia. Conclusion. Vinorelbine demonstrates limited activity in a heavily pretreated group of soft-tissue sarcoma patients. Prospective investigation may be considered for selected sarcoma subtypes.


Assessing the consistency and microbiological effectiveness of household water treatment practices by urban and rural populations claiming to treat their water at home: a case study in Peru.
Household water treatment (HWT) can improve drinking water quality and prevent disease if used correctly and consistently by vulnerable populations. Over 1.1 billion people report treating their water prior to drinking it. These estimates, however, are based on responses to household surveys that may exaggerate the consistency and microbiological performance of the practice-key factors for reducing pathogen exposure and achieving health benefits. The objective of this study was to examine how HWT practices are actually performed by households identified as HWT users, according to international monitoring standards. We conducted a 6-month case study in urban (n = 117 households) and rural (n = 115 households) Peru, a country in which 82.8% of households report treating their water at home. We used direct observation, in-depth interviews, surveys, spot-checks, and water sampling to assess water treatment practices among households that claimed to treat their drinking water at home. While consistency of reported practices was high in both urban (94.8%) and rural (85.3%) settings, availability of treated water (based on self-report) at time of collection was low, with 67.1% and 23.0% of urban and rural households having treated water at all three sampling visits. Self-reported consumption of untreated water in the home among adults and children <5 was common and this was corroborated during home observations. Drinking water of self-reported users was significantly better than source water in the urban setting and negligible but significantly better in the rural setting. However, only 46.3% and 31.6% of households had drinking water <1 CFU/100 mL at all follow-up visits. Our results raise questions about the usefulness of current international monitoring of HWT practices and their usefulness as a proxy indicator for drinking water quality. The lack of consistency and sub-optimal microbiological effectiveness also raises questions about the potential of HWT to prevent waterborne diseases.


ON THE ORIGIN OF HEPARIN : AN EXAMINATION OF THE HEPARIN CONTENT AND THE SPECIFIC CYTOPLASMIC PARTICLES OF NEOPLASTIC MAST CELLS.
1. The spontaneous mast cell tumor of the dog contains heparin. 2. The cytoplasmic particulate content of the tumor mast cells varies with their anaplasia. This conclusion is based on the following findings: (a) in the immature cell of the more malignant tumor the particulate matter appeared in the living cells by phase microscopy to be composed of greyish illdefined particles or as a fine, weakly metachromatic granulation in the fixed and stained preparation; (b) in the mature cells of a relatively benign mast cell tumor, both in the living cell and in stained preparations, the particulate matter occurred in the form of discrete, dense, and strongly metachromatic granules, resembling those of the normal mast cell. 3. The heparin content was large (fifty times that of dog liver) in the growth with mature cells and only moderate (1.7 times) in that with immature cells. 4. Since there may be a great amount of greyish particulate matter (or fine stained granules) in a tumor of relatively low heparin content, it is suggested that this material represents an early or precursor phase in the development of heparin. 5. This possibility and the fact that the blood stream may be invaded by mature tumor mast cells of large heparin content without evident disturbance in the coagulability of the blood suggest the value of a comprehensive biochemical study of the heparin of mast cell tumors.


Application of ancient DNA to the reconstruction of past microbial assemblages and for the detection of toxic cyanobacteria in subtropical freshwater ecosystems.
Ancient DNA (aDNA) analysis of lake sediments is a promising tool for detecting shifts in past microbial assemblages in response to changing environmental conditions. We examined sediment core samples from subtropical, freshwater Laguna Blanca (Uruguay), which has been severely affected by cultural eutrophication since 1960 and where cyanobacterial blooms, particularly those of the saxitoxin-producer Cylindrospermopsis raciborskii, have been reported since the 1990s. Samples corresponding to ~1846, 1852, 2000 and 2007 AD were selected to perform denaturing gradient gel electrophoresis (DGGE) analysis of the 16S-23S rRNA intergenic transcribed spacer (ribosomal ITS) to compare their prokaryotic assemblage composition. Each stratum showed different ITS patterns, but the composition of 21st century samples was clearly different than those of mid-19th century. This compositional change was correlated with shifts in sediment organic matter and chlorophyll a content, which were significantly higher in recent samples. The presence of saxitoxin-producing cyanobacteria was addressed by quantitative real-time PCR of the sxtU gene involved in toxin biosynthesis. This gene was present only in recent samples, for which clone libraries and ITS sequencing indicated the presence of Cyanobacteria. Phylogenetic analyses identified C. raciborskii only in the 2000 sample, shortly after several years when blooms were recorded in the lake. These data suggest the utility of aDNA for the reconstruction of microbial assemblage shifts in subtropical lakes, at least on centennial scales. The application of aDNA analysis to genes involved in cyanotoxin synthesis extends the applicability of molecular techniques in palaeolimnological studies to include key microbial community characteristics of great scientific and social interest.


One-year follow-up study of blood pressure and dietary patterns in dietary approaches to stop hypertension (DASH)-sodium participants.
We conducted a 12-month follow-up study on participants in the Dietary Approaches to Stop Hypertension (DASH)-Sodium trial to determine the impact of the DASH diet and 150, 100, or 50 mmol/day of sodium after discontinuation of the feeding intervention. Upon completion of the trial, 56 of the 113 participants at the clinical center at Duke University entered a longitudinal observational study. Measurements including 24-h ambulatory systolic and diastolic blood pressure (BP), weight, and 24-h urinary electrolytes were obtained at 1, 6, and 12 months after completion of the DASH-Sodium trial. Block food frequency questionnaires were completed at the 12-month visit. Linear mixed-effects regression models were used to analyze the effect of the diet and sodium intake on longitudinal changes in BP during the final feeding period. The DASH participants significantly increased their intakes of fruits or juices and vegetables at 12 months. Control participants had no change in DASH food group intake. Both groups increased sodium intake. Among control participants, systolic and diastolic BP increased 5.33 (95% confidence interval [CI] 0.28, 10.37) and 3.20 mm Hg (95% CI 0.25, 6.16), respectively. Among DASH participants, systolic and diastolic BP increased 3.12 (95% CI -0.44, 6.68) and 0.79 mm Hg (95% CI -1.15, 2.72), respectively. There was a significant effect of the final sodium level by diet on the change in SBP over time (P = .04 for three-way interaction among diet, time of visit, and sodium). After the feeding intervention, DASH diet participants ate more fruits/vegetables and had sustained reductions in BP despite increased sodium intake.


Large Comparative Analyses of Primate Body Site Microbiomes Indicate that the Oral Microbiome Is Unique among All Body Sites and Conserved among Nonhuman Primates.
The study of the mammalian microbiome serves as a critical tool for understanding host-microbial diversity and coevolution and the impact of bacterial communities on host health. While studies of specific microbial systems (e.g., in the human gut) have rapidly increased, large knowledge gaps remain, hindering our understanding of the determinants and levels of variation in microbiomes across multiple body sites and host species. Here, we compare microbiome community compositions from eight distinct body sites among 17 phylogenetically diverse species of nonhuman primates (NHPs), representing the largest comparative study of microbial diversity across primate host species and body sites. Analysis of 898 samples predominantly acquired in the wild demonstrated that oral microbiomes were unique in their clustering, with distinctive divergence from all other body site microbiomes. In contrast, all other body site microbiomes clustered principally by host species and differentiated by body site within host species. These results highlight two key findings: (i) the oral microbiome is unique compared to all other body site microbiomes and conserved among diverse nonhuman primates, despite their considerable dietary and phylogenetic differences, and (ii) assessments of the determinants of host-microbial diversity are relative to the level of the comparison (i.e., intra-/inter-body site, -host species, and -individual), emphasizing the need for broader comparative microbial analyses across diverse hosts to further elucidate host-microbial dynamics, evolutionary and biological patterns of variation, and implications for human-microbial coevolution. IMPORTANCE The microbiome is critical to host health and disease, but much remains unknown about the determinants, levels, and evolution of host-microbial diversity. The relationship between hosts and their associated microbes is complex. Most studies to date have focused on the gut microbiome; however, large gaps remain in our understanding of host-microbial diversity, coevolution, and levels of variation in microbiomes across multiple body sites and host species. To better understand the patterns of variation and evolutionary context of host-microbial communities, we conducted one of the largest comparative studies to date, which indicated that the oral microbiome was distinct from the microbiomes of all other body sites and convergent across host species, suggesting conserved niche specialization within the Primates order. We also show the importance of host species differences in shaping the microbiome within specific body sites. This large, comparative study contributes valuable information on key patterns of variation among hosts and body sites, with implications for understanding host-microbial dynamics and human-microbial coevolution.


Spectrum of critical illness in undocumented border crossers. The Arizona-Mexico border experience.
Approximately 150-250 migrants die each year while attempting to cross the border from Mexico to the Southwest United States. Many border crossers survive the journey, but some develop life-threatening medical complications. Such complications have been subject to little formal analysis. We sought to determine the causes of critical illness in this population and to analyze the hospital course and outcomes of these patients. We retrospectively identified border crossers admitted to the intensive care units (ICUs) of two major teaching hospitals in southern Arizona. We recorded admitting diagnoses, severity of illness, length of stay, resource use, discharge diagnoses, and mortality. Our investigation identified 55 admissions to adult ICUs between January 1, 2010 and December 31, 2012. The median age of patients was 27 years. The median hospital length of stay was 7 days, with a median ICU length of stay of 3 days. The median temperature on arrival to the emergency department was 36.8°C. The most common admission diagnoses included trauma (40), rhabdomyolysis (27), acute liver injury (25), dehydration (24), acute kidney injury (19), and encephalopathy (17). Thirteen patients presented with respiratory failure, six patients with severe sepsis, and two with septic shock. A total of 19 patients required ventilator support during their hospital stay, and 30 required at least one surgical intervention. One patient required renal replacement therapy. The median Acute Physiology and Chronic Health Evaluation II score was 6. All but one patient survived to discharge from the hospital. Border crossers are a unique population of young individuals exposed to high temperatures and extreme conditions. Our review of border crosser admissions showed that most patients demonstrated signs of dehydration and leukocytosis, despite a normal median temperature. The median ICU stay was short, despite a high number of patients requiring ventilator support and surgical intervention. Only one death occurred in this cohort.


Perspectives on the Management of Children in a Biocontainment Unit: Report of the NETEC Pediatric Workgroup.
During the outbreak of Ebola virus disease that struck West Africa during 2014-2016, a small handful of expatriate patients were evacuated to specialized high-level containment care units, or biocontainment units, in the United States and Western Europe. Given the lower mortality rate (18% versus 40% for those treated in Africa) among these patients, it is likely that high-level containment care will be used in the future with increasing frequency. It is also likely that children infected with Ebola and other highly hazardous communicable diseases will someday require such care. The National Ebola Training and Education Center convened a pediatric workgroup to consider the unique and problematic issues posed by these potential child patients. We report here the results of those discussions.


A homological approach to a mathematical definition of pulmonary fibrosis and emphysema on computed tomography.
Three-dimensional imaging is essential to evaluate local abnormalities and understand structure-function relationships in an organ. However, quantifiable and interpretable methods to localize abnormalities remain unestablished. Visual assessments are prone to bias, machine learning methods depend on training images, and the underlying decision principle is usually difficult to interpret. Here, we developed a homological approach to mathematically define emphysema and fibrosis in the lungs on computed tomography (CT). With the use of persistent homology, the density of homological features, including connected components, tunnels, and voids, was extracted from the volumetric CT scans of lung diseases. A pair of CT values at which each homological feature appeared (birth) and disappeared (death) was computed by sweeping the threshold levels from higher to lower CT values. Consequently, fibrosis and emphysema were defined as voxels with dense voids having a longer lifetime (birth-death difference) and voxels with dense connected components having a lower birth, respectively. In an independent dataset including subjects with idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD), and combined pulmonary fibrosis and emphysema (CPFE), the proposed definition enabled accurate segmentation with comparable quality to deep learning in terms of Dice coefficients. Persistent homology-defined fibrosis was closely associated with physiological abnormalities such as impaired diffusion capacity and long-term mortality in subjects with IPF and CPFE, and persistent homology-defined emphysema was associated with impaired diffusion capacity in subjects with COPD. The present persistent homology-based evaluation of structural abnormalities could help explore the clinical and physiological impacts of structural changes and morphological mechanisms of disease progression.NEW & NOTEWORTHY This study proposes a homological approach to mathematically define a three-dimensional texture feature of emphysema and fibrosis on chest computed tomography using persistent homology. The proposed definition enabled accurate segmentation with comparable quality to deep learning while offering higher interpretability than deep learning-based methods.