Computational mapping reveals dramatic effect of Hoogsteen breathing on duplex DNA reactivity with formaldehyde.
Formaldehyde has long been recognized as a hazardous environmental agent highly reactive with DNA. Recently, it has been realized that due to the activity of histone demethylation enzymes within the cell nucleus, formaldehyde is produced endogenously, in direct vicinity of genomic DNA. Should it lead to extensive DNA damage? We address this question with the aid of a computational mapping method, analogous to X-ray and nuclear magnetic resonance techniques for observing weakly specific interactions of small organic compounds with a macromolecule in order to establish important functional sites. We concentrate on the leading reaction of formaldehyde with free bases: hydroxymethylation of cytosine amino groups. Our results show that in B-DNA, cytosine amino groups are totally inaccessible for the formaldehyde attack. Then, we explore the effect of recently discovered transient flipping of Watson-Crick (WC) pairs into Hoogsteen (HG) pairs (HG breathing). Our results show that the HG base pair formation dramatically affects the accessibility for formaldehyde of cytosine amino nitrogens within WC base pairs adjacent to HG base pairs. The extensive literature on DNA interaction with formaldehyde is analyzed in light of the new findings. The obtained data emphasize the significance of DNA HG breathing.


Self-discrepancy and natural killer cell activity: immunological consequences of negative self-evaluation.
The study tested whether self-discrepancy theory could account for changes in natural killer (NK) cell activity after exposure to self-referential stimuli. Anxious, dysphoric, and control Ss were pretested and 1 month later covertly exposed to their own self-guides as well as those of another S. Blood samples were drawn for analysis of NK cytotoxicity and cortisol. The dysphoric Ss manifested the greatest actual:ideal discrepancy, whereas the anxious Ss manifested the greatest actual:ought discrepancy. Content analysis of written responses showed that activating discrepancies induced specific negative states; priming discrepancies also increased cortisol for the anxious Ss. NK activity was lower after self-referential priming for both distressed groups, particularly the anxious Ss. The control Ss showed a trend toward increased NK activity after self-referential priming. The study represents the 1st experimental demonstration that negative self-evaluation can alter immune responses.


Interaction With the Extracellular Matrix Triggers Calcium Signaling in Trypanosoma cruzi Prior to Cell Invasion.
Trypanosoma cruzi, the etiological agent of Chagas disease in humans, infects a wide variety of vertebrates. Trypomastigotes, the parasite infective forms, invade mammalian cells by a still poorly understood mechanism. Adhesion of tissue culture- derived trypomastigotes to the extracellular matrix (ECM) prior to cell invasion has been shown to be a relevant part of the process. Changes in phosphorylation, S-nitrosylation, and nitration levels of proteins, in the late phase of the interaction (2 h), leading to the reprogramming of both trypomastigotes metabolism and the DNA binding profile of modified histones, were described by our group. Here, the involvement of calcium signaling at a very early phase of parasite interaction with ECM is described. Increments in the intracellular calcium concentrations during trypomastigotes-ECM interaction depends on the Ca2+ uptake from the extracellular medium, since it is inhibited by EGTA or Nifedipine, an inhibitor of the L-type voltage gated Ca2+ channels and sphingosine-dependent plasma membrane Ca2+ channel, but not by Vanadate, an inhibitor of the plasma membrane Ca2+-ATPase. Furthermore, Nifedipine inhibits the invasion of host cells by tissue culture- derived trypomastigotes in a dose-dependent manner, reaching 95% inhibition at 100 µM Nifedipine. These data indicate the importance of both Ca2+ uptake from the medium and parasite-ECM interaction for host-cell invasion. Previous treatment of ECM with protease abolishes the Ca2+ uptake, further reinforcing the possibility that these events may be connected. The mitochondrion plays a relevant role in Ca2+ homeostasis in trypomastigotes during their interaction with ECM, as shown by the increment of the intracellular Ca2+ concentration in the presence of Antimycin A, in contrast to other calcium homeostasis disruptors, such as Cyclopiazonic acid for endoplasmic reticulum and Bafilomycin A for acidocalcisome. Total phosphatase activity in the parasite decreases in the presence of Nifedipine, EGTA, and Okadaic acid, implying a role of calcium in the phosphorylation level of proteins that are interacting with the ECM in tissue culture- derived trypomastigotes. In summary, we describe here the increment of Ca2+ at an early phase of the trypomastigotes interaction with ECM, implicating both nifedipine-sensitive Ca2+ channels in the influx of Ca2+ and the mitochondrion as the relevant organelle in Ca2+ homeostasis. The data unravel a complex sequence of events prior to host cell invasion itself.


Helicobacter spp. from gastric biopsies of stranded South American fur seals (Arctocephalus australis).
Gastrointestinal lesions with uncertain etiology have been widely described among pinnipeds. The aim of our study was to investigate the presence of Helicobacter spp. in the gastric mucosa of South American fur seals (Arctocephalusaustralis). Gastric biopsies from thirteen seals, stranded on the shores of the Southwestern Atlantic Ocean in Argentina, were evaluated for the presence of Helicobacter spp. by PCR and DNA sequence analysis. Six gastric biopsies were positive for Helicobacter spp. Pairwise sequence comparisons showed less than 95% identity to novel Helicobacter spp. described from pinnipeds from North America and Australia. However, phylogenetic analysis revealed that the South American fur seal sequences clustered with 99-100% homology with H. cetorum, a species isolated from dolphins and whales. The presence of H. cetorum in pinnipeds, if confirmed by its isolation from the gastric mucosa of these mammals, demonstrates the wide host range of this bacterium in the marine environment.


Perceived Changes in Sexual Interest and Distress About Discrepant Sexual Interest During the First Phase of COVID-19 Pandemic: A Multi-Country Assessment in Cohabiting Partnered Individuals.
Due to COVID-19 pandemic, different restrictive measures in terms of physical distancing and lockdowns have been introduced in most European countries, affecting all facets of social life. Currently, little is known about how partnered individuals perceive changes in their sexual life during this complex emergency. This study explored retrospectively assessed changes in sexual interest for one's partner and levels of distress related to perceived sexual interest discrepancy during the first phase of the pandemic in a large-scale online sample of partnered individuals (n = 4813; Mage = 38.5 years, SD = 10.74) recruited between May and July 2020 in seven European Union countries and Turkey. We also examined the possible role of approach/avoidance motives for sex in reported changes in sexual interest and associated distress. Most participants (53%) reported no change in their sexual interest during the pandemic, followed by those who reported an increase (28.5%). The pattern was similar across the eight countries. Distress about discrepant sexual interest, which was only weakly related to changes in sexual interest, was significantly associated with relationship quality and emotional closeness with a partner, coping with and worrying about the pandemic, and specific motivation for sex. In contrast to avoidant and relationship-focused approach motivation, ego-focused approach motivation was related to stable sexual interest during the pandemic. The current study contributes to the understanding of the link between sexual interest and complex emergencies. Considering that the COVID-19 pandemic continues, the reported experiences and perceptions are prone to change.


Biochemical variables from Holstein-Friesian calves older than one week are comparable to those obtained from adult animals of stable metabolic status on the same farm.
Age-related changes in blood variables have previously been reported for cattle. A few studies have investigated differences in biochemistry analytes over time using repeat measures. The purpose of the study was to investigate the variation in select variables during the first 3 months of life of Holstein-Friesian calves using repeat sampling. Twenty healthy calves were sampled on the day of birth (0) and at 6, 13, 21, 30, 60, and 90 days of life. Analyzed variables included concentration of glucose, total protein, albumin, urea and creatinine, and activity of AST, GGT, and CK. Reference intervals (RI) were obtained from 40 adult cows using specific computer software. The variation in each analyte throughout the study period and differences relative to the RI in adult cattle were assessed. With the exception of glucose, the studied variables were affected by the calf's age. However, for most of the analytes studied, the values obtained from calves from one week of age onwards remained within the RI determined in adult cattle. During the calves' first week of life, some variables outside the RI of adult cattle were due to the changes associated with birth and colostrum intake. With the exception of AST and GGT activity, RI for standard biochemistry variables determined for adult cattle are likely to be valid for calves older than one week. However, special consideration is necessary for calves within their first week of life.


Magnetic resonance imaging preprocessing and radiomic features for classification of autosomal dominant polycystic kidney disease genotype.
Our study aims to investigate the impact of preprocessing on magnetic resonance imaging (MRI) radiomic features extracted from the noncystic kidney parenchyma of patients with autosomal dominant polycystic kidney disease (ADPKD) in the task of classifying PKD1 versus PKD2 genotypes, which differ with regard to cyst burden and disease outcome. The effect of preprocessing on radiomic features was investigated using a single T2-weighted fat saturated (T2W-FS) MRI scan from PKD1 and PKD2 subjects (29 kidneys in total) from the Consortium for Radiologic Imaging Studies of Polycystic Kidney Disease study. Radiomic feature reproducibility using the intraclass correlation coefficient (ICC) was computed across MRI normalizations (z-score, reference-tissue, and original image), gray-level discretization, and upsampling and downsampling pixel schemes. A second dataset for genotype classification from 136 subjects T2W-FS MRI images previously enrolled in the HALT Progression of Polycystic Kidney Disease study was matched for age, gender, and Mayo imaging classification class. Genotype classification was performed using a logistic regression classifier and radiomic features extracted from (1) the noncystic kidney parenchyma and (2) the entire kidney. The area under the receiver operating characteristic curve (AUC) was used to evaluate the classification performance across preprocessing methods. Radiomic features extracted from the noncystic kidney parenchyma were sensitive to preprocessing parameters, with varying reproducibility depending on the parameter. The percentage of features with good-to-excellent ICC scores ranged from 14% to 58%. AUC values ranged between 0.47 to 0.68 and 0.56 to 0.73 for the noncystic kidney parenchyma and entire kidney, respectively. Reproducibility of radiomic features extracted from the noncystic kidney parenchyma was dependent on the preprocessing parameters used, and the effect on genotype classification was sensitive to preprocessing parameters. The results suggest that texture features may be indicative of genotype expression in ADPKD.


Comparative analysis of European bat lyssavirus 1 pathogenicity in the mouse model.
European bat lyssavirus 1 is responsible for most bat rabies cases in Europe. Although EBLV-1 isolates display a high degree of sequence identity, different sublineages exist. In individual isolates various insertions and deletions have been identified, with unknown impact on viral replication and pathogenicity. In order to assess whether different genetic features of EBLV-1 isolates correlate with phenotypic changes, different EBLV-1 variants were compared for pathogenicity in the mouse model. Groups of three mice were infected intracranially (i.c.) with 102 TCID50/ml and groups of six mice were infected intramuscularly (i.m.) with 105 TCID50/ml and 102 TCID50/ml as well as intranasally (i.n.) with 102 TCID50/ml. Significant differences in survival following i.m. inoculation with low doses as well as i.n. inoculation were observed. Also, striking variations in incubation periods following i.c. inoculation and i.m. inoculation with high doses were seen. Hereby, the clinical picture differed between general symptoms, spasms and aggressiveness depending on the inoculation route. Immunohistochemistry of mouse brains showed that the virus distribution in the brain depended on the inoculation route. In conclusion, different EBLV-1 isolates differ in pathogenicity indicating variation which is not reflected in studies of single isolates.


RASSF1A Enhances Chemosensitivity of NSCLC Cells Through Activating Autophagy by Regulating MAP1S to Inactivate Keap1-Nrf2 Pathway.
Cisplatin (DDP) is an effective first-line therapy for non-small cell lung cancer (NSCLC) treatment; however, it can cause resistance and thus pose an obstacle to the efficacy of chemotherapy in NSCLC. This study aims to detect the effect of RASSF1A on DDP resistance of NSCLC and the underlying mechanism. The expression levels of RASSF1A and microtubule-associated protein 1S (MAP1S) were investigated by qRT-PCR and Western blot and their interaction was testified by co-immunoprecipitation (Co-IP) analysis. The IC50 value of DDP on A549 and A549/DDP cells (DDP-resistant cells) was measured. A549/DDP cells were transfected with pCDNA3.1-RASSF1A, pCDNA3.1-MAP1S, or si-RASSF1A, followed by treated with DDP. Cell counting kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EDU) were employed to measure cell survival rate. Western blot was applied to test the levels of autophagy-associated proteins p62, LC3II, and LC3I. Immunofluorescence staining was used to detect the green fluorescent protein (GFP)-LC3 puncta to evaluate the level of autophagy. Finally, a xenograft model in nude mice using A549/DDP cells was developed. RASSF1A and MAP1S were lowly expressed and positively correlated in NSCLC tissues. We observed that RASSF1A and MAP1S overexpression significantly enhanced DDP-induced effects in A549 and A549/DDP cells, including decreased cell viability, as well as increased autophagy levels. Besides, investigations into the mechanism between RASSF1A and MAP1S disclosed that RASSF1A could regulate MAP1S to inactivate the Keap1-Nrf2 pathway, thus activating autophagy to enhance chemosensitivity. Moreover, consistent results were confirmed in vivo experiments. RASSF1A increases chemosensitivity in NSCLC by facilitating autophagy via MAP1S-mediated Keap1-Nrf2 pathway.


Norovirus outbreak amid COVID-19 in the United Kingdom; priorities for achieving control.
Norovirus, an enteric virus primarily responsible for gastroenteritis outbreaks worldwide, is currently causing outbreaks around the United Kingdom during the COVID-19 pandemic. With an already exhausted health care system, the significant burden norovirus can have on the National Health Service, including economic and social burdens, is immense and cannot be tolerated. Primary challenges and priorities to be focused on due to the increase in norovirus outbreaks include a further depletion of health care services, increase cases in schools, nurseries, and care facilities, underreporting of the cases, and no effective vaccine being available. Therefore, it is essential to increase awareness about norovirus and its transmission in public, take necessary precautions, and increase reporting of cases. This article discusses the impact norovirus has during the COVID-19 pandemic, and the challenges, and recommendations to achieve control before it reaches epidemic levels.


Synthesis and photocatalytic performance of titanium dioxide nanofibers and the fabrication of flexible composite films from nanofibers.
Titanium dioxide nanofibers were synthesized and applied in flexible composite films that are easy to handle and recycle after use. The nanofibers were obtained in a multi-step procedure. First, sodium titanate nanofibers were prepared from TiO2 nanoparticles through the alkali hydrothermal method. In the next step, sodium hydrogen titanate nanofibers were made by washing the sodium titanate nanofibers in HCl solution. Finally, the sodium hydrogen titanate nanofibers were transformed to TiO2 anatase nanofibers by calcination in air. The photocatalytic activity of TiO2 anatase nanofibers were evaluated and compared to a TiO2 nanoparticle catalyst by decomposing methyl orange dye in aqueous solutions. The achieved reaction rate constant of TiO2 anatase nanofibers was comparable to that of Degussa P25. Paper-like flexible composite films were prepared by co-filtrating aqueous dispersions of TiO2 catalyst materials and cellulose. The composite films made from the nanofibers exhibit better mechanical integrity than those of the nanoparticle-cellulose composites.


A molecular Debye-Hückel theory of solvation in polar fluids: An extension of the Born model.
A dielectric response theory of solvation beyond the conventional Born model for polar fluids is presented. The dielectric response of a polar fluid is described by a Born response mode and a linear combination of Debye-Hückel-like response modes that capture the nonlocal response of polar fluids. The Born mode is characterized by a bulk dielectric constant, while a Debye-Hückel mode is characterized by its corresponding Debye screening length. Both the bulk dielectric constant and the Debye screening lengths are determined from the bulk dielectric function of the polar fluid. The linear combination coefficients of the response modes are evaluated in a self-consistent way and can be used to evaluate the electrostatic contribution to the thermodynamic properties of a polar fluid. Our theory is applied to a dipolar hard sphere fluid as well as interaction site models of polar fluids such as water, where the electrostatic contribution to their thermodynamic properties can be obtained accurately.


A Tool-free Neuronavigation Method based on Single-view Hand Tracking.
This work presents a novel tool-free neuronavigation method that can be used with a single RGB commodity camera. Compared with freehand craniotomy placement methods, the proposed system is more intuitive and less error prone. The proposed method also has several advantages over standard neuronavigation platforms. First, it has a much lower cost, since it doesn't require the use of an optical tracking camera or electromagnetic field generator, which are typically the most expensive parts of a neuronavigation system, making it much more accessible. Second, it requires minimal setup, meaning that it can be performed at the bedside and in circumstances where using a standard neuronavigation system is impractical. Our system relies on machine-learning-based hand pose estimation that acts as a proxy for optical tool tracking, enabling a 3D-3D pre-operative to intra-operative registration. Qualitative assessment from clinical users showed that the concept is clinically relevant. Quantitative assessment showed that on average a target registration error (TRE) of 1.3cm can be achieved. Furthermore, the system is framework-agnostic, meaning that future improvements to hand-tracking frameworks would directly translate to a higher accuracy.


The function of p120 catenin in filopodial growth and synaptic vesicle clustering in neurons.
At the developing neuromuscular junction (NMJ), physical contact between motor axons and muscle cells initiates presynaptic and postsynaptic differentiation. Using Xenopus nerve-muscle cocultures, we previously showed that innervating axons induced muscle filopodia (myopodia), which facilitated interactions between the synaptic partners and promoted NMJ formation. The myopodia were generated by nerve-released signals through muscle p120 catenin (p120ctn), a protein of the cadherin complex that modulates the activity of Rho GTPases. Because axons also extend filopodia that mediate early nerve-muscle interactions, here we test p120ctn's function in the assembly of these presynaptic processes. Overexpression of wild-type p120ctn in Xenopus spinal neurons leads to an increase in filopodial growth and synaptic vesicle (SV) clustering along axons, whereas the development of these specializations is inhibited following the expression of a p120ctn mutant lacking sequences important for regulating Rho GTPases. The p120ctn mutant also inhibits the induction of axonal filopodia and SV clusters by basic fibroblast growth factor, a muscle-derived molecule that triggers presynaptic differentiation. Of importance, introduction of the p120ctn mutant into neurons hinders NMJ formation, which is observed as a reduction in the accumulation of acetylcholine receptors at innervation sites in muscle. Our results suggest that p120ctn signaling in motor neurons promotes nerve-muscle interaction and NMJ assembly.


Quality of life in children with epilepsy: How does it compare with the quality of life in typical children and children with cerebral palsy?
Our objective was to compare the quality of life (QoL) of children with epilepsy to that of typical children and children with cerebral palsy (CP). We measured self- and proxy-reported QoL of children with epilepsy and contrasted that with data for typical children (European KIDSCREEN project) and children with CP (SPARCLE study). Children ages 8-12 years with epilepsy were recruited from six Canadian sites. Same-aged children with CP and children in the general population aged 8-11 years came from several European countries. All participants completed the KIDSCREEN-52 questionnaire. Our results showed no clinically important differences (>0.5 SD) between self-reported QoL in 345 children with epilepsy compared with 489 children with CP or 5950 children in the general population. However, parents reported clinically important differences between the epilepsy and the other groups in five KIDSCREEN-52 domains. Compared with the CP group, parents of children with epilepsy reported better QoL in physical well-being (Cohen d=0.81), social support (d=0.80), and autonomy (d=0.72). Parents reported poorer QoL in the domains of mood and emotions compared with both contrast groups (d=-0.72 and d=-0.53), and in the domain of bullying compared with the CP group (d=-0.51). Families should find comfort in the results, which indicate that children with epilepsy do not perceive any important differences in QoL compared with their typical peers. The comparisons of parental reports detect their group-specific observations and worries that need to be addressed by the health-care providers and may require specifically designed assessment batteries followed by appropriate interventions.


Ranges of phenotypic flexibility in healthy subjects.
A key feature of metabolic health is the ability to adapt upon dietary perturbations. A systemic review defined an optimal nutritional challenge test, the "PhenFlex test" (PFT). Recently, it has been shown that the PFT enables the quantification of all relevant metabolic processes involved in maintaining or regaining homeostasis of metabolic health. Furthermore, it was demonstrated that quantification of PFT response was more sensitive as compared to fasting markers in demonstrating reduced phenotypic flexibility in metabolically impaired type 2 diabetes subjects. This study aims to demonstrate that quantification of PFT response can discriminate between different states of health within the healthy range of the population. Therefore, 100 healthy subjects were enrolled (50 males, 50 females) ranging in age (young, middle, old) and body fat percentage (low, medium, high), assuming variation in phenotypic flexibility. Biomarkers were selected to quantify main processes which characterize phenotypic flexibility in response to PFT: flexibility in glucose, lipid, amino acid and vitamin metabolism, and metabolic stress. Individual phenotypic flexibility was visualized using the "health space" by representing the four processes on the health space axes. By quantifying and presenting the study subjects in this space, individual phenotypic flexibility was visualized. Using the "health space" visualization, differences between groups as well as within groups from the healthy range of the population can be easily and intuitively assessed. The health space showed a different adaptation to the metabolic PhenFlex test in the extremes of the recruited population; persons of young age with low to normal fat percentage had a markedly different position in the health space as compared to persons from old age with normal to high fat percentage. The results of the metabolic PhenFlex test in conjunction with the health space reliably assessed health on an individual basis. This quantification can be used in the future for personalized health quantification and advice.


A cosmesis outcome substudy in a prospective, randomized trial comparing radioguided seed localization with standard wire localization for nonpalpable, invasive, and in situ breast carcinomas.
The primary study objective was to compare the cosmetic result of radioguided seed localization (RSL) with wire localization (WL). A subgroup of patients enrolled in a multicentered, randomized trial comparing WL with RSL participated. Frontal photographs were taken 1 and 3 years postsurgery. The European Organization for Research and Treatment of Cancer Cosmetic Rating System was used to evaluate cosmesis outcomes by the patient and a panel of 5 raters. The study enrolled 73 patients (WL, n = 38; RSL, n = 35). Most patients rated their overall cosmesis as "excellent" or "good" (76% WL, 80% RSL). Patient and panel ratings on all cosmetic outcomes were similar between groups. Multivariable regression for overall cosmesis found larger specimen volume and reoperation to be predictors of worse ratings. All cosmetic outcomes assessed were similar after WL and RSL. The comparable outcomes may reflect similar reoperation rates and volumes of excision between groups.


Quantification of Silymarin in Silybi mariani fructus: Challenging the Analytical Performance of Benchtop vs. Handheld NIR Spectrometers on Whole Seeds.
The content of the flavonolignan mixture silymarin and its individual components (silichristin, silidianin, silibinin A, silibinin B, isosilibinin A, and isosilibinin B) in whole and milled milk thistle seeds (Silybi mariani fructus) was analyzed with near-infrared spectroscopy. The analytical performance of one benchtop and two handheld near-infrared spectrometers was compared. Reference analysis was performed with HPLC following a Soxhlet extraction (European Pharmacopoeia) and a more resource-efficient ultrasonic extraction. The reliability of near-infrared spectral analysis determined through partial least squares regression models constructed independently for the spectral datasets obtained by the three spectrometers was as follows. The benchtop device NIRFlex N-500 performed the best both for milled and whole seeds with a root mean square error of CV between 0.01 and 0.17%. The handheld spectrometer MicroNIR 2200 as well as the microPHAZIR provided a similar performance (root mean square error of CV between 0.01 and 0.18% and between 0.01 and 0.23%, respectively). We carried out quantum chemical simulation of near-infrared spectra of silichristin, silidianin, silibinin, and isosilibinin for interpretation of the results of spectral analysis. This provided understanding of the absorption regions meaningful for the calibration. Further, it helped to better separate how the chemical and physical properties of the samples affect the analysis. While the study demonstrated that milling of samples slightly improves the performance, it was deemed to be critical only for the analysis carried out with the microPHAZIR. This study evidenced that rapid and nondestructive quantification of silymarin and individual flavonolignans is possible with miniaturized near-infrared spectroscopy in whole milk thistle seeds.


Simulating retention of rare alleles in small populations to assess management options for species with different life histories.
Preserving allelic diversity is important because it provides the capacity for adaptation and thus enables long-term population viability. Allele retention is difficult to predict in animals with overlapping generations, so we used a new computer model to simulate retention of rare alleles in small populations of 3 species with contrasting life-history traits: North Island Brown Kiwi (Apteryx mantelli; monogamous, long-lived), North Island Robins (Petroica longipes; monogamous, short-lived), and red deer (Cervus elaphus; polygynous, moderate lifespan). We simulated closed populations under various demographic scenarios and assessed the amounts of artificial immigration needed to achieve a goal of retaining 90% of selectively neutral rare alleles (frequency in the source population = 0.05) after 10 generations. The number of immigrants per generation required to meet the genetic goal ranged from 11 to 30, and there were key similarities and differences among species. None of the species met the genetic goal without immigration, and red deer lost the most allelic diversity due to reproductive skew among polygynous males. However, red deer required only a moderate rate of immigration relative to the other species to meet the genetic goal because nonterritorial breeders had a high turnover. Conversely, North Island Brown Kiwi needed the most immigration because the long lifespan of locally produced territorial breeders prevented a large proportion of immigrants from recruiting. In all species, the amount of immigration needed generally decreased with an increase in carrying capacity, survival, or reproductive output and increased as individual variation in reproductive success increased, indicating the importance of accurately quantifying these parameters to predict the effects of management. Overall, retaining rare alleles in a small, isolated population requires substantial investment of management effort. Use of simulations to explore strategies optimized for the populations in question will help maximize the value of this effort..


Evaluation of the Becton Dickinson Directigen test for respiratory syncytial virus in nasopharyngeal aspirates.
A premarket trial of the Becton Dickinson Directigen respiratory syncytial virus membrane-based enzyme immunoassay compared the test with virus isolation for the detection of respiratory syncytial virus in 583 nasopharyngeal aspirates. After modification, the Directigen test showed a sensitivity of 83% and a specificity of 90%. It offers the potential for an efficient bedside test--without the need for any equipment--for the diagnosis of respiratory syncytial virus infection and requires only a 0.25-ml sample volume. However, for optimum reliability, freezing-thawing of samples and access to a confirmatory test were shown to be necessary.


The role of sulfoacetaldehyde sulfo-lyase in the mineralization of isethionate by an environmental Acinetobacter isolate.
An environmental Acinetobacter isolate, strain ICD, utilized isethionate at concentrations up to at least 20 mM as carbon and energy source, with essentially quantitative sulfate accumulation. The initial step in isethionate metabolism is likely to be its oxidation to sulfoacetaldehyde since inducible sulfoacetaldehyde sulfo-lyase activity was demonstrated in isethionate-grown cells by in vitro assay and gel zymography; sulfoacetaldehyde itself did not induce the enzyme. Isethionate-grown cells of Acinetobacter sp. ICD, unlike those of most other C-S bond-cleaving strains described, also contained an inducible sulfite-oxidizing activity. The results provide further evidence that sulfoacetaldehyde sulfo-lyase plays a central role in the mineralization of biogenic sulfonates.


CD8+ T-cell-Mediated Immunoediting Influences Genomic Evolution and Immune Evasion in Murine Gliomas.
Cancer immunoediting shapes tumor progression by the selection of tumor cell variants that can evade immune recognition. Given the immune evasion and intratumor heterogeneity characteristic of gliomas, we hypothesized that CD8+ T cells mediate immunoediting in these tumors. We developed retrovirus-induced PDGF+ Pten -/- murine gliomas and evaluated glioma progression and tumor immunogenicity in the absence of CD8+ T cells by depleting this immune cell population. Furthermore, we characterized the genomic alterations present in gliomas that developed in the presence and absence of CD8+ T cells. Upon transplantation, gliomas that developed in the absence of CD8+ T cells engrafted poorly in recipients with intact immunity but engrafted well in those with CD8+ T-cell depletion. In contrast, gliomas that developed under pressure from CD8+ T cells were able to fully engraft in both CD8+ T-cell-depleted mice and immunocompetent mice. Remarkably, gliomas developed in the absence of CD8+ T cells exhibited increased aneuploidy, MAPK pathway signaling, gene fusions, and macrophage/microglial infiltration, and showed a proinflammatory phenotype. MAPK activation correlated with macrophage/microglia recruitment in this model and in the human disease. Our studies indicate that, in these tumor models, CD8+ T cells influence glioma oncogenic pathways, tumor genotype, and immunogenicity. This suggests immunoediting of immunogenic tumor clones through their negative selection by CD8+ T cells during glioma formation.


Two-kernel image deconvolution.
A method for deconvolution of the true image of an object from two recorded images is proposed. These two images have to be made by an imaging system with two different but interconnected kernels. The method is formulated as a system of Fredholm equations of the first kind reduced to a single functional equation in Fourier space. The kernels of the system and the true image of an object are found from the same recorded images.


Cancer mortality in relation to measures of occupational exposure to crocidolite at Wittenoom Gorge in Western Australia.
The separate and combined effects of duration and intensity of exposure to crocidolite on mortality from lung cancer, malignant mesothelioma, and stomach cancer were examined in 6506 male former crocidolite miners and millers at Wittenoom Gorge, Western Australia. Each subject who had died from lung cancer (92), mesothelioma (31), or stomach cancer (17) was matched with up to 20 control subjects of the same age who were not known to have died before the index subject. Relations of dose and time of exposure to crocidolite to risk of death were modelled by conditional logistic regression. For lung cancer, the best fitting multiplicative model was one which estimated a relative risk (RR) of 1.12 (95% CI 1.04-1.20) per year of exposure and 1.01 (95% CI 1.00-1.01) per fibre/ml. This was statistically indistinguishable from an additive model showing an increase in RR of 0.01045 (95% CI 0.008-0.020) per f/ml year. For mesothelioma the best fitting model appeared to be one estimating a RR of 24.9 (95% CI 3.51-1.77) per log year since first exposed and a RR of 10.5 (95% CI 3.12-35.1) if exposed for longer than six months. This was not distinguishable statistically from a model that showed mortality increasing as the fourth power of time since first exposed less the fourth power of time since last exposed. The effect of intensity of exposure on the RR for mesothelioma was only slight. There was no consistent effect of any measure of exposure to crocidolite on death from stomach cancer.


Elevated autophagic sequestration of mitochondria and lipid droplets in steatotic hepatocytes of chronic ethanol-treated rats: an immunohistochemical and electron microscopic study.
Ethanol-induced hepatic steatosis may induce the progression of alcoholic liver disease. The involvement of autophagic clearance of damaged mitochondria (mitophagy) and lipid droplets (LDs) (lipophagy) in chronic ethanol-induced hepatic steatosis is not clearly understood. Adult Wistar rats were fed either 5 % ethanol in Lieber-DeCarli liquid diet or an isocaloric control diet for 10 weeks. Light microscopy showed marked steatosis in hepatocytes of ethanol-treated rats (ETRs), which was further revealed by transmission electron microscopy (TEM), where significant numbers of large LDs and damaged mitochondria were detected in steatotic hepatocytes. Moreover, TEM demonstrated that hepatocyte steatosis was associated with greatly enhanced autophagic vacuole (AV) formation compared to control hepatocytes. Mitochondria and LDs were the predominant contents of AVs in steatotic hepatocytes. Immunohistochemistry of LC3, a specific marker of early AVs (autophagosomes), demonstrated an extensive punctate pattern in hepatocytes of ETRs, while LC3 puncta were much less frequent in control hepatocytes. This was confirmed by immunoelectron microscopy (IEM), which showed localization of LC3 to autophagosomes sequestering damaged mitochondria and LDs. In addition, IEM revealed that PINK1 (a sensor of mitochondrial damage and marker of mitophagy) was overexpressed in mitochondria of ETRs. Enhanced autophagic lysosomal activity was evidenced by increased immunolabeling of LAMP-2, a marker of late AVs (autolysosomes) in hepatocytes of ETRs and colocalization of LC3 and lysosomal cathepsins using double immunofluorescence labeling. Increased AVs in hepatocytes of ETRs reflect ethanol toxicity and could represent a possible protective mechanism via stimulation of mitophagy and lipophagy.


Cytosine base modifications regulate DNA duplex stability and metabolism.
DNA base modifications diversify the genome and are essential players in development. Yet, their influence on DNA physical properties and the ensuing effects on genome metabolism are poorly understood. Here, we focus on the interplay of cytosine modifications and DNA processes. We show by a combination of in vitro reactions with well-defined protein compositions and conditions, and in vivo experiments within the complex networks of the cell that cytosine methylation stabilizes the DNA helix, increasing its melting temperature and reducing DNA helicase and RNA/DNA polymerase speed. Oxidation of methylated cytosine, however, reverts the duplex stabilizing and genome metabolic effects to the level of unmodified cytosine. We detect this effect with DNA replication and transcription proteins originating from different species, ranging from prokaryotic and viral to the eukaryotic yeast and mammalian proteins. Accordingly, lack of cytosine methylation increases replication fork speed by enhancing DNA helicase unwinding speed in cells. We further validate that this cannot simply be explained by altered global DNA decondensation, changes in histone marks or chromatin structure and accessibility. We propose that the variegated deposition of cytosine modifications along the genome regulates DNA helix stability, thereby providing an elementary mechanism for local fine-tuning of DNA metabolism.


Calculation of genetic risks in Duchenne muscular dystrophy by geneticists in the United Kingdom.
Pedigrees of Duchenne muscular dystrophy were sent to 32 geneticists in the United Kingdom, asking for their calculations for genetic risks for female relatives. Eighteen participated, of whom only 7 gave correct answers for all pedigrees. The method most used for calculations (the 'Bayesian' method) was most liable to errors in new situations. An alternative, simpler method is presented.


Distinctive epigenomic alterations in NF1-deficient cutaneous and plexiform neurofibromas drive differential MKK/p38 signaling.
Benign peripheral nerve sheath tumors are the clinical hallmark of Neurofibromatosis Type 1. They account for substantial morbidity and mortality in NF1. Cutaneous (CNF) and plexiform neurofibromas (PNF) share nearly identical histology, but maintain different growth rates and risk of malignant conversion. The reasons for this disparate clinical behavior are not well explained by recent genome or transcriptome profiling studies. We hypothesized that CNFs and PNFs are epigenetically distinct tumor types that exhibit differential signaling due to genome-wide and site-specific methylation events. We interrogated the methylation profiles of 45 CNFs and 17 PNFs from NF1 subjects with the Illumina EPIC 850K methylation array. Based on these profiles, we confirm that CNFs and PNFs are epigenetically distinct tumors with broad differences in higher-order chromatin states and specific methylation events altering genes involved in key biological and cellular processes, such as inflammation, RAS/MAPK signaling, actin cytoskeleton rearrangement, and oxytocin signaling. Based on our identification of two separate DMRs associated with alternative leading exons in MAP2K3, we demonstrate differential RAS/MKK3/p38 signaling between CNFs and PNFs. Epigenetic reinforcement of RAS/MKK/p38 was a defining characteristic of CNFs leading to pro-inflammatory signaling and chromatin conformational changes, whereas PNFs signaled predominantly through RAS/MEK. Tumor size also correlated with specific CpG methylation events. Taken together, these findings confirm that NF1 deficiency influences the epigenetic regulation of RAS signaling fates, accounting for observed differences in CNF and PNF clinical behavior. The extension of these findings is that CNFs may respond differently than PNFs to RAS-targeted therapeutics raising the possibility of targeting p38-mediated inflammation for CNF treatment.


Towards living well with young onset dementia: An exploration of coping from the perspective of those diagnosed.
There is increasing recognition that dementia in people under 65 years represents a unique challenge to sufferers and services alike. However, by either ignoring this population or sampling them together with older adults, current research has failed to reflect this. This study explored the specific experience of living and coping with Alzheimer's disease in younger life from the perspective of those diagnosed. Semi-structured interviews were analysed using interpretive phenomenological analysis, and four themes were identified: 'disruption of the life-cycle', 'identity', 'social orientation' and 'agency'. The experience of living and coping with young onset Alzheimer's disease was strongly situated within an individual's social context. Most significantly, participants felt too young to develop the disease and felt out of time with age-related psychological tasks. Coping strategies that attempted to redress and normalise the life-cycle were identified. The contributions of the study are discussed in the context of the human development literature.


Comparison of high-intensity laser therapy and ultrasound treatment in the patients with lumbar discopathy.
The aim of the present study was to evaluate the efficiency of high intensity laser and ultrasound therapy in patients who were diagnosed with lumbar disc herniation and who were capable of performing physical exercises. 65 patients diagnosed with lumbar disc were included in the study. The patients were randomly divided into three groups: Group 1 received 10 sessions of high intensity laser to the lumbar region, Group 2 received 10 sessions of ultrasound, and Group 3 received medical therapy for 10 days and isometric lumbar exercises. The efficacy of the treatment modalities was compared with the assessment of the patients before the therapy at the end of the therapy, and in third month after the therapy. Comparing the changes between groups, statically significant difference was observed in MH (mental health) parameter before treatment between Groups 1 and 2 and in MH parameter and VAS score in third month of the therapy between Groups 2 and 3. However, the evaluation of the patients after ten days of treatment did not show significant differences between the groups compared to baseline values. We found that HILT, ultrasound, and exercise were efficient therapies for lumbar discopathy but HILT and ultrasound had longer effect on some parameters.


Variant PNLDC1, Defective piRNA Processing, and Azoospermia.
P-element-induced wimpy testis (PIWI)-interacting RNAs (piRNAs) are short (21 to 35 nucleotides in length) and noncoding and are found almost exclusively in germ cells, where they regulate aberrant expression of transposable elements and postmeiotic gene expression. Critical to the processing of piRNAs is the protein poly(A)-specific RNase-like domain containing 1 (PNLDC1), which trims their 3' ends and, when disrupted in mice, causes azoospermia and male infertility. We performed exome sequencing on DNA samples from 924 men who had received a diagnosis of nonobstructive azoospermia. Testicular-biopsy samples were analyzed by means of histologic and immunohistochemical tests, in situ hybridization, reverse-transcriptase-quantitative-polymerase-chain-reaction assay, and small-RNA sequencing. Four unrelated men of Middle Eastern descent who had nonobstructive azoospermia were found to carry mutations in PNLDC1: the first patient had a biallelic stop-gain mutation, p.R452Ter (rs200629089; minor allele frequency, 0.00004); the second, a novel biallelic missense variant, p.P84S; the third, two compound heterozygous mutations consisting of p.M259T (rs141903829; minor allele frequency, 0.0007) and p.L35PfsTer3 (rs754159168; minor allele frequency, 0.00004); and the fourth, a novel biallelic canonical splice acceptor site variant, c.607-2A→T. Testicular histologic findings consistently showed error-prone meiosis and spermatogenic arrest with round spermatids of type Sa as the most advanced population of germ cells. Gene and protein expression of PNLDC1, as well as the piRNA-processing proteins PIWIL1, PIWIL4, MYBL1, and TDRKH, were greatly diminished in cells of the testes. Furthermore, the length distribution of piRNAs and the number of pachytene piRNAs was significantly altered in men carrying PNLDC1 mutations. Our results suggest a direct mechanistic effect of faulty piRNA processing on meiosis and spermatogenesis in men, ultimately leading to male infertility. (Funded by Innovation Fund Denmark and others.).


Carotid plaque, intima-media thickness, and incident aortic stenosis: a prospective cohort study.
Aortic stenosis (AS) shares risk factors with atherosclerotic vascular disease. Carotid intima-media thickness (IMT) and plaque may reflect the cumulative damage from exposure to different atherosclerotic risk factors. We examined the relationship of carotid IMT and plaque with incident AS in a prospective population-based study. A random sample of participants (age, 45-68 years) in the population-based Malmö Diet and Cancer Study underwent B-mode ultrasound with measurements of IMT and the presence of plaque in the common carotid artery (n=5079). Potential risk factors for incident AS were studied in age- and sex-adjusted and expanded multivariable-adjusted Cox regression models. A total of 69 (1.4%) participants developed AS during up to 20 years of follow-up. Significant risk factors for AS in age- and sex-adjusted analyses were (P<0.05) body mass index, low-density lipoprotein cholesterol, hypertension, diabetes mellitus, smoking, C-reactive protein, plaque, and IMT. In contrast, high-density lipoprotein cholesterol, triglycerides, height, and leukocyte count were not significantly associated with AS (P>0.05). After adjustments, IMT, plaque, age, smoking, C-reactive protein, low-density lipoprotein cholesterol, and diabetes mellitus remained significantly associated with incident AS. IMT was no longer significantly associated with AS after adjustments for plaque and systolic blood pressure, but plaque remained significantly associated with incident AS. Traditional cardiovascular risk factors were individually associated with incident AS, and in multivariable models low-density lipoprotein cholesterol, smoking, age, presence of plaque, C-reactive protein, and diabetes mellitus remained significant predictors of incident AS. AS represents a vascular disorder related to carotid plaque, with potential implications for the pathophysiology and prevention of this disease.


[Implementing the seven roles of the specialist training reform].
On 1 January 2004, a new medical specialist reform was implemented. The Danish School of Education, University of Aarhus, conducted research from 2002-2006 in the form of two studies to study educational practice within medical specialist training programmes. This article, based on selected data from the second study, sheds light on the reception of the reform's seven competence roles. The roles represented a pedagogical paradigm shift which calls for a new interpretation of the spectrum of medical expertise. However, the reform's reception has not previously been subject to qualitative analysis. The qualitative data consists of two semi-structured focus group interviews comprising 13 doctors and six rounds of observation of three junior hospital doctors with individual follow-up interviews. The study shows that, almost two years after the reform's introduction, the roles were not ascribed equal value by the doctors, even if this was intended by the reform. The description of competence objectives does not sufficiently reflect everyday practice where competences are often socially situated and constructed. Furthermore, the study indicates a need for training of supervisors in the evaluation of the competence objectives. The doctors' perception of the roles indicate a discrepancy between their official formulation and everyday practice. This discrepancy can partly be attributed to the profession's focus on the role as medical expert, inexhaustive descriptions of objectives, and a surfeit of pedagogical qualifications among instructors.


Simultaneous extraction and fractionation of polycyclic aromatic hydrocarbons and their oxygenated derivatives in soil using selective pressurized liquid extraction.
In this study, a selective pressurized liquid extraction (PLE) method which can extract polycyclic aromatic hydrocarbons (PAHs) and their oxygenated derivatives (oxy-PAHs) from contaminated soil and simultaneously separate them into two fractions was developed. The method uses extraction cells packed with a chromatographic adsorbent and extraction solvents of increasing polarity. Several experiments were conducted on both spiked and authentic contaminated soil samples. Different types of adsorbents, combinations of extraction solvents, and extraction temperatures were tested in order to find a method that could fulfill the purpose of the study. The final method was based on extraction cells packed with 2% deactivated silica gel. The PAHs were extracted with cyclohexane/dichloromethane (9:1) at 120 degrees C, after which the oxy-PAHs where extracted with cyclohexane/dichloromethane (1:3) at 150 degrees C. The PAHs and oxy-PAHs were efficiently separated into two fractions, and only trace amounts of some compounds were found in the inappropriate fraction. The recoveries of the PAHs were mostly above 70% and of the oxy-PAHs, above 90%. The linearity of the method was good, and the calibration curves for most compounds had a regression coefficient better than 0.99 and an intercept close to the origin of coordinates. When the selective PLE method was applied to seven authentic soil samples, the results were found to be in good agreement with those of a reference method based on Soxhlet extraction and silica gel cleanup and also in good agreement with the certified reference values available for one of the soils. The selective PLE method is faster and consumes less solvent than a traditional method based on separate extraction and fractionation steps. The selective PLE method is, therefore, suitable for the concurrent analysis of PAHs and oxy-PAHs during large-scale soil contamination studies. This will provide more information about the soil contamination and the levels of toxicity than an ordinary PAH analysis.


Lessons from COVID-19 for Pandemic Preparedness: Proceedings from a Multi-Stakeholder Think Tank.
While the Coronavirus Disease 2019 (COVID-19) pandemic continues to present global challenges, sufficient time has passed to reflect on lessons learned and use those insights to inform policy and approaches to prepare for the next pandemic. In May 2022, the Duke Clinical Research Institute (DCRI) convened a Think Tank with thought-leaders from academia, clinical practice, the pharmaceutical industry, patient advocacy, the National Institutes of Health, the US Food and Drug Administration (FDA), and Centers for Disease Control and Prevention (CDC), to share firsthand, expert knowledge of the insights gained from the COVID-19 pandemic and how this acquired knowledge can help inform the next pandemic response. The Think Tank focused on pandemic preparedness, therapeutics, vaccines, and challenges related to clinical trial design and scale-up during the early phase of a pandemic. Based on the multi-faceted discussions, we outline ten key steps to an improved and equitable pandemic response.


[Squamous Cell Carcinoma of the Conjunctiva]
The squamous cell carcinoma of the conjunctiva is a neoplasia of the ocular surface that may present with different macroscopic patterns. We present the case of a 75-year-old man that comes presenting this tumor for a long time, involving in an important way its vision.


Single-dose intranasal vaccination elicits systemic and mucosal immunity against SARS-CoV-2.
A safe and durable vaccine is urgently needed to tackle the COVID19 pandemic that has infected >15 million people and caused >620,000 deaths worldwide. As with other respiratory pathogens, the nasal compartment is the first barrier that needs to be breached by the SARS-CoV-2 virus before dissemination to the lung. Despite progress at remarkable speed, current intramuscular vaccines are designed to elicit systemic immunity without conferring mucosal immunity. We report the development of an intranasal subunit vaccine that contains the trimeric or monomeric spike protein and liposomal STING agonist as adjuvant. This vaccine induces systemic neutralizing antibodies, mucosal IgA responses in the lung and nasal compartments, and T-cell responses in the lung of mice. Single-cell RNA-sequencing confirmed the concomitant activation of T and B cell responses in a germinal center-like manner within the nasal-associated lymphoid tissues (NALT), confirming its role as an inductive site that can lead to long-lasting immunity. The ability to elicit immunity in the respiratory tract has can prevent the initial establishment of infection in individuals and prevent disease transmission across humans.


The balance between feeling and knowing: affective and cognitive empathy are reflected in the brain's intrinsic functional dynamics.
Affective empathy (AE) is distinguished clinically and neurally from cognitive empathy (CE). While AE is selectively disrupted in psychopathy, autism is associated with deficits in CE. Despite such dissociations, AE and CE together contribute to normal human empathic experience. A dimensional measure of individual differences in AE 'relative to' CE captures this interaction and may reveal brain-behavior relationships beyond those detectable with AE and CE separately. Using resting-state fMRI and measures of empathy in healthy adults, we show that relative empathic ability (REA) is reflected in the brain's intrinsic functional dynamics. Dominance of AE was associated with stronger functional connectivity among social-emotional regions (ventral anterior insula, orbitofrontal cortex, amygdala, perigenual anterior cingulate). Dominance of CE was related to stronger connectivity among areas implicated in interoception, autonomic monitoring and social-cognitive processing (brainstem, superior temporal sulcus, ventral anterior insula). These patterns were distinct from those observed with AE and CE separately. Finally, REA and the strength of several functional connections were associated with symptoms of psychopathology. These findings suggest that REA provides a dimensional index of empathic function and pathological tendencies in healthy adults, which are reflected in the intrinsic functional dynamics of neural systems associated with social and emotional cognition.


[The Mitsuda reaction and its antigen, good subjects for a bibliography].
Some references of the history of Mitsuda reaction and its antigen, as well as for the results and significance of this unique reaction are presented. In spite of the great advances of modern immunology. Mitsuda reaction is still the best method for the diagnosis of the forms of hanseniasis and for the prognosis of the disease in patients and in healthy persons.


Sleep quality trajectories from head and neck cancer diagnosis to six months after treatment.
Patients with head and neck cancer (HNC) often report disturbances in their sleep quality, impairing their quality of life. This study aims to examine the trajectories of sleep quality from diagnosis up to 6-month after treatment, as well as the pre-treatment risk factors for poor sleep trajectories. Sleep quality (Pittsburgh sleep quality index) was measured shortly after diagnosis (pre-treatment), and at 3 and 6 months after finishing treatment. Patients were categorized into 5 trajectory groups. We examined the association of sleep quality trajectories with sociodemographic and clinical characteristics, coping style, HNC symptoms, and psychological distress. Among 412 included patients, about a half either had a persistent good sleep (37.6%) or an improving (16.5%) trajectory. About a third had a persistent poor sleep (21.8%) or worsening (10.9%) sleep trajectory. The remaining patients (13.1%), alternated between good and poor sleep. Using persistent good sleep as a reference outcome, persistent poor sleepers were more likely to be woman (odds ratio [OR] = 1.98, 95% confidence interval [CI] 1.01-3.90), use painkillers prior to treatment (OR = 2.52, 95% CI 1.33-4.77), and have more pre-treatment anxiety symptoms (OR = 1.26, 95% CI 1.15-1.38). Unfavorable sleep quality trajectories are prevalent among HNC patients from pre-treatment to 6-month after treatment. A periodic sleep evaluation starting shortly after HNC diagnosis is necessary to identify persistent sleep problems, especially among high-risk group.


B7 expression on T cells down-regulates immune responses through CTLA-4 ligation via T-T interactions [corrections].
Although B7 on APCs has a well-recognized role in T cell costimulation, little is known about the functional significance of constitutive and activation-induced B7 expression that also occurs on T cells. To analyze the role of B7 on T cells, B7-1/B7-2-deficient mice (B7 double knockout) and mice overexpressing B7-2 exclusively on T cells (B7-2 transgenic) were used as T cell donors for allogeneic transplant recipients, and graft-vs-host disease (GVHD) was assessed. B7 double-knockout T cells resulted in significant GVHD acceleration compared with wild-type T cells. Conversely, B7-2 transgenic donor T cells mediated reduced GVHD mortality compared with wild-type T cells. Data indicated that B7 expression on T cells down-regulated alloresponses through CTLA-4 ligation. This study is the first to provide definitive in vivo data illustrating the importance of T cell-associated B7 as a negative regulator of immune responses in a clinically relevant murine model of GVHD. The up-regulation of B7 on T cells may be an important component of normal immune homeostasis.


Distribution of [3H]diisopropylfluorophosphate, [3H]soman, [3H]sarin, and their metabolites in mouse brain.
The brain area distribution of [3H]diisopropylfluorophosphate, [3H]soman, and [3H]sarin and their metabolites in mice was studied after iv administration of sublethal doses. At the appropriate time after the injection of the radiolabeled organophosphate, the mice were decapitated and their brains were dissected into seven areas. There was a relatively even distribution of the parent compounds and their metabolites in all brain areas except the hypothalamus, which contained concentrations of parent compounds the metabolites that were 2-5 times greater than those in other brain areas. Concentrations of the parent compounds and free metabolites declined steadily throughout the time course, whereas concentrations of the bound metabolites remained relatively constant between 6 and 24 hr. There was no correlation between the disposition of soman, and its metabolites, and cholinesterase inhibition in brain areas, which implicates other central mechanisms in the production of organophosphate effects. However, the higher concentrations of organophosphates and their metabolites in the hypothalamus suggest that this area might be important with respect to the pharmacological effects or the toxicity of these compounds.


The Association of Intracranial Pressure Monitoring and Mortality: A Propensity Score-Matched Cohort of Isolated Severe Blunt Traumatic Brain Injury.
Intracranial pressure (ICP) monitoring in traumatic brain injury (TBI) is common. Yet, its efficacy varies between studies, and the actual effect on the outcome is debated. This study investigates the association of ICP monitoring and clinical outcome in patients with an isolated severe blunt TBI. Patients were recruited from the American College of Surgeons-Trauma Quality Improvement Program database during 2014. Inclusion criteria were limited to adult patients (≥18 years) who had a sustained isolated severe intracranial injury (Abbreviated Injury Scale [AIS] head of ≥3 and Glasgow Coma Scale [GCS] of ≤8) following blunt trauma to the head. Patients with AIS score >0 for any extracranial body area were excluded. Patients' demographics, injury characteristics, interventions, and outcomes were collected for analysis. Patients receiving ICP monitoring were matched in a 1:1 ratio with controls who were not ICP monitored using propensity score matching. A total of 3289 patients met inclusion criteria. Of these, 601 (18.3%) were ICP monitored. After propensity score matching, 557 pairs were available for analysis with a mean age of 44 (standard deviation 18) years and 80.2% of them were male. Median GCS on admission was 4[3,7], and a third of patients required neurosurgical intervention. There were no statistical differences in any variables included in the analysis between the ICP-monitored group and their matched counterparts. ICP-monitored patients required significantly longer intensive care unit and hospital length of stay and had an increased mortality risk with odds ratio of 1.6 (95% confidence interval: 1.1-2.5, P = 0.038). ICP monitoring is associated with increased in-hospital mortality in patients with an isolated severe TBI. Further investigation into which patients may benefit from this intervention is required.


BAALC and ERG expression levels are associated with outcome and distinct gene and microRNA expression profiles in older patients with de novo cytogenetically normal acute myeloid leukemia: a Cancer and Leukemia Group B study.
BAALC and ERG expression levels are prognostic markers in younger (< 60 years) cytogenetically normal acute myeloid leukemia (CN-AML) adults; their prognostic impact in older (≥ 60 years) patients requires further investigation. We evaluated pretreatment expression of BAALC and ERG in 158 de novo patients treated on cytarabine/daunorubicin-based protocols. The patients were also characterized for other established molecular prognosticators. Low BAALC and ERG expression levels were associated with better outcome in univariable and multivariable analyses. Expression levels of both BAALC and ERG were the only factors significantly associated with overall survival upon multivariable analysis. To gain biological insights, we derived gene expression signatures associated with BAALC and ERG expression in older CN-AML patients. Furthermore, we derived the first microRNA expression signatures associated with the expression of these 2 genes. In low BAALC expressers, genes associated with undifferentiated hematopoietic precursors and unfavorable outcome predictors were down-regulated, whereas HOX genes and HOX-gene-embedded microRNAs were up-regulated. Low ERG expressers presented with down-regulation of genes involved in the DNA-methylation machinery, and up-regulation of miR-148a, which targets DNMT3B. We conclude that in older CN-AML patients, low BAALC and ERG expression associates with better outcome and distinct gene and microRNA expression signatures that could aid in identifying new targets and novel therapeutic strategies for older patients.


Repair of calvarial bone defects in mice using electrospun polystyrene scaffolds combined with β-TCP or gold nanoparticles.
Non-biodegradable porous polystyrene (PS) scaffolds, composed of microfibers, have been prepared by electrospinning for the reconstruction of large bone defects. PS microfibers were prepared by incorporating β-TCP grains inside the polymer or grafting gold nanoparticles surface functionalized with mercaptosuccinic acid. Cytocompatibility of the three types of scaffolds (PS, β-TCP-PS and Au-PS) was studied by seeding human mesenchymal stem cells. Biocompatibility was evaluated by implanting β-TCP-PS and Au-PS scaffolds into a critical size (4mm) calvarial defect in mice. Calvaria were taken 6, 9, and 12 weeks after implantation; newly formed bone and cellular response was analyzed by microcomputed tomography (microCT) and histology. β-TCP-PS scaffolds showed a significantly higher cell proliferation in vitro than on PS or Au-PS alone; clearly, the presence of β-TCP grains improved cytocompatibility. Biocompatibility study in the mouse calvaria model showed that β-TCP-PS scaffolds were significantly associated with more newly-formed bone than Au-PS. Bone developed by osteoconduction from the defect margins to the center. A dense fibrous connective tissue containing blood vessels was identified histologically in both types of scaffolds. There was no inflammatory foci nor giant cell in these areas. AuNPs aggregates were identified histologically in the fibrosis and also incorporated in the newly-formed bone matrix. Although the different types of PS microfibers appeared cytocompatible during the in vitro experiment, they appeared biotolerated in vivo since they induced a fibrotic reaction associated with newly formed bone.


Microfluidics-based rapid measurement of nitrite in human blood plasma.
Nitric oxide (NO) is one of the vital gasotransmitters that takes part in many biological pathways such as infection, inflammation and ischemia, immune response, neurotransmission, and cardiovascular systems. Nitrite is one of the primary metabolites of NO and is considered to be a circulating storage pool for NO. Here, we report direct and rapid measurement of nitrite in human blood plasma using a fluorescence-based microfluidic method. The study revealed the factors that affect the endogenous concentration of nitrite in blood plasma, mainly the presence of blood cells, hemoglobin, and soluble proteins. We find that separation of blood plasma immediately after sample collection and subsequent dilution of plasma with buffer at a ratio of 1 : 4 eliminates the interference from cells and proteins, providing reliable measurements. The proposed method can measure plasma nitrite in the concentration range of 0-20 µM with a limit of detection of 60 nM and a sensitivity of 5.64 µM-1 within 10 min of sample collection. By spiking nitrite into plasma, a linear correlation between the nitrite concentration and FL intensity is obtained, which is utilized further to measure the endogenous concentration of nitrite present in the plasma of healthy volunteers and patients. The study revealed that the endogenous nitrite concentration in the blood plasma of healthy humans falls in the range of 0.4-1.2 µM. Furthermore, the study with blood samples obtained from patients showed that nitrite levels are inversely correlated with the total cholesterol and low-density lipoproteins levels, which is in good agreement with the literature.


Incarceration and recidivism among sexual offenders.
The relationship between incarceration and recidivism was investigated in a sample of 627 adult male sexual offenders. Incarceration for the index offense was unrelated to sexual or violent recidivism. This was the case whether incarceration was examined as a dichotomous variable (incarceration vs. community sentence) or as a continuous variable (length of incarceration). Risk for sexual recidivism was assessed with a modified version of the Rapid Risk Assessment for Sexual Offense Recidivism. There was no evidence that the relationship between incarceration and recidivism was confounded or moderated by risk or that length of incarceration and recidivism were non-linearly associated. Sentencing sexual offenders to terms of incarceration appears to have little, if any, impact on sexual and violent recidivism following release.


A Multireference Ab Initio Study of the Diradical Isomers of Pyrazine.
Three diradical pyrazine isomers were characterized using highly correlated, multireference methods. The lowest lying singlet and triplet state geometries of 2,3-didehydropyrazine ( ortho), 2,5-didehydropyrazine ( para), and 2,6-didehydropyrazine ( meta) were determined. Two active reference spaces were utilized. The complete active space (CAS) (8,8) includes the σ and σ* orbitals on the dehydrocarbon atoms as well as the valence π and π* orbitals. The CAS (12,10) reference space includes two additional orbitals corresponding to the in-phase and out-of-phase nitrogen lone pair orbitals. Adiabatic and vertical gaps between the lowest lying singlet and triplet states, optimized geometries, canonicalized orbital energies, unpaired electron densities, and spin polarization effects were compared. We find that the singlet states of each diradical isomer contain two significantly weighted configurations, and the larger active space is necessary for the proper physical characterization of both the singlet and triplet states. The singlet-triplet splitting is very small for the 2,3-didehydropyrazine ( ortho) and 2,6-didehydropyrazine ( meta) isomers (+1.8 and -1.4 kcal/mol, respectively) and significant for the 2,5-didehydropyrazine ( para) isomer (+28.2 kcal/mol). Singlet geometries show through-space interactions between the dehydocarbon atoms in the 2,3-didehydropyrazine ( ortho) and 2,6-didehydropyrazine ( meta) isomers. An analysis of the effectively unpaired electrons suggests that the 2,5-didehydropyrazine ( para) isomer also displays through-bond coupling between the diradical electrons.


Laser treatment of upper gastrointestinal hemorrhage.
The goal of laser treatment of the visible vessel or bleeding ulcer is to stop the bleeding to allow healing of the ulcer base. Adjuvant techniques including the use of vasoconstrictive agents, tissue adhesives, and tissue injection hold future promise in complementing laser therapy. Laser photocoagulation appears to be an effective method of treatment. At this time, however, the therapy of this serious clinical problem is evolving. As experience increases and as new technology and techniques are applied, the development of an effective treatment for the bleeding peptic ulcer or visible vessel is even more promising.


Destabilization of the metal site as a hub for the pathogenic mechanism of five ALS-linked mutants of copper, zinc superoxide dismutase.
Amyotrophic lateral sclerosis (ALS) is a lethal neurodegenerative disease, with no effective pharmacological treatment. Its pathogenesis is unknown, although a subset of the cases is linked to genetic mutations. A significant fraction of the mutations occur in one protein, copper, zinc superoxide dismutase (SOD1). The toxic function of mutant SOD1 has not been elucidated, but damage to the metal site of the protein is believed to play a major role. In this work, we study the electrostatic loop of SOD1, which we had previously proposed to work as a "solvent seal" isolating the metal site from water molecules. Out of the five contact points identified between the electrostatic loop and its dock in the rest of the protein, three points were found to be affected by ALS-linked mutations, with a total of five mutations identified. The effect of the five mutations was studied using methods of computational chemistry. We found that four of the mutations destabilize the proposed solvent seal, while the fifth mutation directly affects the metal-site stability. In the two contact points unaffected by ALS-linked mutations, the side chains of the residues were not found to play a stabilizing role. Our results show that the docking of the electrostatic loop to the rest of SOD1 plays a role in ALS pathogenesis, in support of that structure acting as a solvent barrier for the metal site. The results provide a unified pathogenic mechanism for five different ALS-linked mutations of SOD1.


Alterations in vitamin A and thyroid hormone status in anorexia nervosa and associated disorders.
Vitamin A and thyroid hormone status was investigated in 27 patients with anorexia nervosa. Subjects were divided into three groups based on eating behavior and serum carotene concentrations: anorexic (dietary restriction), normal carotene; anorexic, elevated serum carotene; bulimic, elevated serum carotene. All bulimic subjects fulfilling selection criteria were hypercarotenemic (weight loss and reduced metabolic rate). Data were compared to normal healthy volunteers. Serum retinol and retinol-binding protein levels were normal in all subjects whereas retinyl esters were elevated in the hypercarotenemic groups. Hypercarotenemia was primarily a result of elevation of vitamin A active carotenoids, especially beta-carotene. Diet was excluded from the etiology of hypercarotenemia. Thyroid hormones T4 and T3 were significantly depressed in hypercarotenemic groups and rT3 increased. A concomitant alteration in vitamin-hormone status is observed with progressive metabolic alterations: low T3, T4, and elevated retinyl esters in subjects with the hypercarotenemia associated with anorexia nervosa.


Hodgkin disease-derived cell lines expressing ubiquitous mitochondrial creatine kinase show growth inhibition by cyclocreatine treatment independent of apoptosis.
Ubiquitous mitochondrial creatine kinase (uMtCK), a key enzyme in energy metabolism, was identified by differential display PCR to be specifically overexpressed in L1236, the first cell line of definite Hodgkin origin. RT-PCR confirmed overexpression of uMtCK in the L1236 cell line and the absence of cytosolic B-CK, which is co-expressed with MtCK physiologically. Cyclocreatine (cCr), whose phosphorylated form is a very poor substrate for CK, inhibited proliferation of the L1236 cell line nearly entirely. This inhibition by cCr was partially reversed by competition with creatine, which by itself had no effect on proliferation of the L1236 cell line. Although these results support a role of CK activity in the inhibitory action of cCr, it remains open whether the cCr effect is due to its inhibition of CK-linked energy metabolism or if alternative mechanisms have to be considered. Because the anti-proliferative effect of cCr was not due to induction of apoptosis, in contrast to most other anticancer agents, treatment with the creatine analogue cCr may represent an advantageous therapeutic approach for cells resistant to programmed cell death.


Inner canthal distance and golden proportion as predictors of maxillary central incisor width in south Indian population.
Even though the constant relation of golden proportion and inner canthal distance (ICD) with the width of the maxillary central incisor (CIW) has been found in European population, it may not be applied to Indian population as we differ from Europeans racially and genetically. Hence, this study was carried out with the objectives of determining if these parameters are applicable to our population also. Three hundred south Indian subjects between 18 and 26 years of age, free from facial and dental deformities were examined. Inner canthus of each eye was used as soft tissue landmark. The maxillary central incisors were measured at the contact point area with the help of digital vernier caliper. The CIW was also calculated using golden proportion ratio to obtain the calculated central incisor width. A comparison was made with measured width. Statistical analyses were done to identify any significant difference using "Z" tests. Pearson's Correlation Coefficient test was used to evaluate the measured and the calculated width of the central incisor. ICD and the width of two maxillary incisors were in golden proportion in south Indian population. Also, ICD when multiplied by a decreasing function value of the golden proportion and divided by 2 is a reliable predictor of determining CIW. As in the European population, the ICD and the golden proportion are reliable predictors for determining the width of the maxillary central incisors in the south Indian population also.


Optic chiasm astrocytomas of childhood. 1. Long-term follow-up.
We reviewed 61 patients seen from 1975 to 1994 with a biopsy-confirmed optic chiasm astrocytoma or a characteristic clinical and imaging presentation. The mean age at diagnosis was 72 months, including 30 who were less than 5 years old. The mean tumor diameter at presentation was 31 mm among 54 tumors measured. Tumors involved the optic nerve in 41, hypothalamus in 38, optic tract in 15, and optic radiations in 2. Four patients, all with neurofibromatosis, received no treatment. Forty patients received conventional radiation therapy at a mean age of 89 months. Chemotherapy was given to 19 children and 7 of these were followed for greater than 1 year. Six of these 7 had progressive disease which required subsequent surgery and/or radiation therapy. Thirty-six patients underwent surgery. We restricted our analysis of outcome to those 38 patients followed for longer than 60 months. Development was normal in 13%, mildly disabled in 53%, severely disabled in 21% and dead in 13%. Children were severely disabled or dead in 47% if younger than 5 years at diagnosis, and 21% if older. There was no endocrine dysfunction in 18% and new endocrine failure was seen in 61%. More posterior tumors did not portend a poorer prognosis. Children less than 5 years old had a poorer prognosis. Neurofibromatosis did not have a marked effect on outcome. Intracranial hypertension was an indicator of a poor prognosis. No specific treatment modality was clearly superior in terms of long-term survival or improvement in quality of life.


Ischaemic preconditioning of skeletal muscle. 1. Protection against the structural changes induced by ischaemia/reperfusion injury.
Ischaemic preconditioning is a process by which exposure of a tissue to a short period of non-damaging ischaemic stress leads to resistance to the deleterious effects of a subsequent prolonged ischaemic stress. It has been extensively described in the heart, but few studies have examined the possibility that it can occur in skeletal muscle. We have used a rat model of ischaemia of one limb to examine this possibility. Exposure of the hind limb to a period of ischaemia of five minutes and reperfusion for five minutes significantly protected the tibialis anterior muscle against the structural damage induced by a subsequent period of limb ischaemia for four hours and reperfusion for one hour. This protection was evident on examination of the muscle by both light and electron microscopy. Longer or shorter times of prior ischaemia had no effect.


Cognitive impairment and dietary habits among elders: the Velestino Study.
To investigate the association of dietary habits with cognitive function among elders (>65 years). Complete sociodemographic, dietary information, serum measurements, and Mini-Mental State Examination (MMSE) assessments were available for 237 elderly men and 320 women residing in Velestino, Greece (a rural Greek town). All models were adjusted for age, education, social activity, smoking, depression symptomatology (using the Geriatric Depression Scale), MedDietScore (range 0-55), and metabolic syndrome. About 49.8% men and 66.6% women had MMSE scores <24, with a mean MMSE score of 22.7±4.43 and 21.1±4.73, respectively. Adherence to the Mediterranean diet was moderate (mean MedDietScore of 34.1±3.25 in men and 35.1±2.48 in women). Indicative cognitive impairment (MMSE score <24) was positively associated with age and low education in women and with depressive symptoms, low education status, and low social activity in men. Adherence to the Mediterranean diet was positively associated with MMSE score in men (P=.02), but inversely associated in women (P=.04). Concerning the food groups studied, intake of pulses, nuts, and seeds was associated with lower likelihood of having MMSE score<24 in men (P=.04). Only the Mediterranean dietary pattern showed a significant association with MMSE score positive for cognitive impairment (i.e., protective in men, but not in women), while individual food groups or nutrients did not achieve significance. The latter findings support the role of whole diet in the prevention of mental disorders, and state a research hypothesis for a sex-diet interaction on cognitive function among elders.


FERMENTATIVE AND PHOTOCHEMICAL PRODUCTION OF HYDROGEN IN ALGAE.
1.. After 2 hours of fermentation in nitrogen the metabolism of those algae which were found capable of photoreduction with hydrogen changes in such a way that molecular hydrogen is released from the cell in addition to carbon dioxide. 2. The amount of hydrogen formed anaerobically in the dark depends on the amount of some unknown reserve substance in the cell. More hydrogen is formed in presence of added glucose, but no proportionality has been found between the amount of substrate added and that of hydrogen formed. This is probably due to the fact that two types of fermentation reactions exist, with little or no connection between them. Whereas mainly unknown organic acids are formed during the autofermentation, the addition of glucose causes a considerable increase in the production of lactic acid. 3. Algae which have been fermenting for several hours in the dark produce upon illumination free hydrogen at several times the rate observed in the dark, provided carbon dioxide is absent. 4. Certain concentrations of dinitrophenol strongly inhibit the evolution of hydrogen in the dark. Fermentation then continues mainly as a reaction leading to lactic acid. In such poisoned algae the photochemical liberation of hydrogen still continues. 5. If the algae are poisoned with dinitrophenol the presence of carbon dioxide will not interfere with the photochemical evolution of hydrogen. 6. The amount of hydrogen released in this new photochemical reaction depends on the presence of an unknown hydrogen donor in the cell; it can be increased by the addition of glucose but not in proportion to the amount added. 7. The results obtained allow for a more correct explanation of the anaerobic induction period previously described for Scenedesmus and similar algae. The possibility of a photochemical evolution of hydrogen had not been taken into account in the earlier experiments. 8. The origin of the hydrogen released under the influence of light is discussed.


Socio-economic and food system drivers of nutrition and health transitions in The Gambia from 1990 to 2017.
In common with many nations undergoing a nutrition transition, micronutrient deficiencies, undernutrition and overnutrition coexist in The Gambia. Addressing these challenges simultaneously would require transformational changes in the country's food system. However, the evidence base that would enable informed decision-making in the Gambian food system has been scant, despite several sources of routinely-collected data being available. This descriptive study brings together data from four open-access global databases on food supply, political, economic, and demographic variables, and nutrition and health between 1990 and 2017 to study potential leverage points for improvement in the food system. It compares trends in food supply and nutritional outcomes in The Gambia against regional and global averages, and identifies potential drivers taken from a food systems framework. The data show that, over the past three decades, total energy supply has increased, and obesity is rising quickly, but iron deficiency persists in a proportion of the population. Overall diet composition is poor, with lower availability of fruit and vegetables and higher supply of sugar and oils compared to regional and global averages. Domestic production is low for most food groups and so a high dependence on imports from other countries bridges the gap in terms of energy supply. Measures of economic development, particularly GDP, were positively related with supply of cereals and animal source foods over time, but no such relationship was observed with fruit and vegetable supply. Food system policy to improve nutrition and health outcomes in The Gambia needs to focus on improving the diversity of food supply - especially fruit and vegetables - and maximizing national domestic production to reduce reliance on food imports. The use of open-source global datasets can be feasible in exploring food system characteristics and trends at the national level and could be applied in other contexts.


[Use of the protoplast fusion and regeneration method for screening antibiotic producers among inactive strains of Streptomyces].
Intraspecies fusion of protoplasts of two strains of Streptomyces fradiae, i.e native protoplasts of an inactive strain INA 00708 and heat inactivated protoplasts of a neomycin-producing strain ATCC 10745, and regeneration of the protoplasts of the inactive strain INA 00708 resulted in formation of clones producing neomycin and clones synthesizing antibiotics of an unknown nature differing from neomycin. All the active clones were unstable and lost their antibiotic activity in subcultures. Regeneration of the protoplasts of 4 different inactive strains of Streptomyces sp. also resulted in formation of active clones which were unstable and lost their capacity for the antibiotic synthesis after the first subculture. The data in principal indicate to the possible use of protoplast fusion and regeneration in screening of cultures producing new antibiotics among inactive strains of streptomycetes. However, the efficiency of such procedures is low since the experiments are labor-consuming and the resulting active clones are genetically unstable.


Application of artificial neural network models in occupational safety and health utilizing ordinal variables.
Safety professionals and practitioners are always searching for methods to accurately assess the association between exposures and possible occupational disorders or diseases and predict the outcome of any variable. Statistical analysis and logistic regression (LR) in particular are among the most popular tools being used today. Artificial neural network (ANN) models are another method of predicting outcomes, which are gradually finding their way into the safety field. Limited studies have shown that they are capable of predicting outcomes more accurately than LR, but they have been tested either on continuous or on dichotomous variables or combinations of them. The objective of this research was to demonstrate that ANN models can perform better than LR models with data sets comprised of all ordinal variables, which has not been done so far. The data set used in this research was collected from construction workers using the Work Compatibility questionnaire. The data set contained only ordinal variables both as input (exposure) and as output (outcome) variables. LR models and ANN models were constructed using the same data set and the performance of all models was compared by using the log-likelihood ratio. The result of this study showed that ANN models performed significantly better than LR models with a data set of all ordinal variables as well as other types of variables such as dichotomous and continuous.


Gender specific factors associated with having stopped smoking among in-school adolescents in Ukraine: results from the Global Youth Tobacco Survey 2005.
The prevalence of cigarette smoking in Ukraine is different between genders and is among the highest in the world. There is need to identify gender-specific factors that are associated with having stopped smoking among adolescents. We used data from the Ukraine Global Youth Tobacco Survey 2005. We carried out a backward stepwise logistic regression analysis with having stopped smoking as the outcome.Altogether, 2800 adolescents reported having ever smoked cigarettes. Overall 64.1% (63.4% male, and 65.5% female) adolescents reported having stopped smoking. Male adolescents who stated that smoking decreases body weight were 25% more likely, while female adolescents were 9% less likely to stop smoking. While male adolescents who received support on how to stop smoking from a family member were 7% less likely, female adolescents were 60% more likely to stop smoking. Furthermore, while male adolescents who received a lecture on the harmful effects of smoking were 10% less likely, female adolescents were 9% more likely to stop smoking. Finally both male and female adolescents who were sure or most probably that they would not smoke a cigarette offered to them by their best friends were more likely, and those adolescents who were sure that smoking is harmful to health were less likely to stop smoking. Our study has identified some factors that are associated with having quit smoking that are gender-specific. We believe public health programs targeting adolescent smoking should consider these factors in their design and implementation of gender sensitive interventions.


Fetal dextrocardia: diagnosis and outcome in two tertiary centres.
To evaluate the incidence of fetal dextrocardia, associated cardiac and extracardiac malformations, and outcome. Retrospective echocardiographic study. Two tertiary centres for fetal cardiology. 81 consecutive fetuses with a fetal dextrocardia presenting at Guy's Hospital, London, between 1983 and 2003 and at Hôpital Robert Debré, Paris, between 1988 and 2003. Fetal dextrocardia was defined as a condition in which the major axis of the heart points to the right. The incidence was 0.22%. There were 38 fetuses (47%) with situs solitus (SS), 24 (30%) with situs ambiguus (SA), and 19 (23%) with situs inversus (SI). Structural cardiac malformations were found in 25 cases (66%) of SS, 23 cases (96%) of SA, and 12 cases (63%) of SI. Extracardiac malformations were identified in 12 cases (31%) of SS, in five cases (21%) of SA, and in two cases (10%) of SI. Of the 81 cases of fetal dextrocardia, there were 27 interrupted pregnancies (15 of 24 SA, 10 of 38 SS, and 2 of 19 SI), six intrauterine deaths (3 of 38 SS, 2 of 24 SA, and 1 of 19 SI), and five neonatal deaths (3 of 24 SA, 1 of 19 SI, and 1 of 38 SS). There were 43 survivors (24 of 38 SS, 15 of 19 SI, and 4 of 24 SA). The majority of fetuses with dextrocardia referred for fetal echocardiography have associated congenital heart disease. There is a broad spectrum of cardiac malformation and the incidence varies according to the atrial situs. Fetal echocardiography enables detection of complex congenital heart disease so that parents can be appropriately counselled.


Corynebacterium sepsis in oncology patients. Predisposing factors, diagnosis, and treatment.
Bacteremia due to a newly described species of Corynebacterium was identified in 12 patients during a two-year period. Infection occurred after a long period of hospitalization, extended granulocytopenia, and treatment with several antibiotics. Breaks in the mucocutaneous surfaces were the origin of infection in eight patients. All patients had evidence of colonization reported by cultures before infection. The invading organism caused death in three patients. Vancomycin hydrochloride was the antibiotic of choice. Corynebacterium is increasingly recognized as a primary cause of morbidity and mortality of patients with tumors.


Oil palm vegetation liquor: a new source of phenolic bioactives.
Waste from agricultural products represents a disposal liability, which needs to be addressed. Palm oil is the most widely traded edible oil globally, and its production generates 85 million tons of aqueous by-products annually. This aqueous stream is rich in phenolic antioxidants, which were investigated for their composition and potential in vitro biological activity. We have identified three isomers of caffeoylshikimic acid as major components of oil palm phenolics (OPP). The 2,2-diphenyl-1-picrylhydrazyl assay confirmed potent free radical scavenging activity. To test for possible cardioprotective effects of OPP, we carried out in vitro LDL oxidation studies as well as ex vivo aortic ring and mesenteric vascular bed relaxation measurements. We found that OPP inhibited the Cu-mediated oxidation of human LDL. OPP also promoted vascular relaxation in both isolated aortic rings and perfused mesenteric vascular beds pre-contracted with noradrenaline. To rule out developmental toxicity, we performed teratological studies on rats up to the third generation and did not find any congenital anomalies. Thus, these initial studies suggest that OPP is safe and may have a protective role against free radical damage, LDL oxidation and its attendant negative effects, as well as vascular constriction in mitigating atherosclerosis. Oil palm vegetation liquor thus represents a new source of phenolic bioactives.


Deferoxamine posttreatment reduces ischemic brain injury in neonatal rats.
Iron catalyzes the formation of damaging reactive species during cerebral reperfusion. Brain iron concentration is highest at birth, so the brain of the asphyxiated newborn may be at increased risk of iron-dependent injury. We investigated whether the ferric iron chelator deferoxamine could reduce hypoxic-ischemic brain injury in neonatal rats. Because deferoxamine has concentration-dependent activities other than iron chelation, we measured brain deferoxamine levels and calculated deferoxamine pharmacokinetic parameters. We produced hypoxic-ischemic injury to the right cerebral hemisphere of 7-day-old rats by right common carotid artery ligation followed by 2.25 hours of hypoxia in 8% oxygen. At 5 minutes of recovery from hypoxia the rats received 100 mg/kg deferoxamine mesylate or saline subcutaneously. Rats (saline, n = 33; deferoxamine, n = 38) were killed at 42 hours of recovery to assess early acute edema by measurement of hemispheric water content. Other rats (saline, n = 31; deferoxamine, n = 32) were killed at 30 days of age for morphometric determination of right hemisphere atrophy. In still other rats, we measured deferoxamine levels in blood and brain after hypoxia-ischemia. Deferoxamine significantly reduced right hemisphere injury as measured by early water content (P < .01) and later atrophy (P = .019). Deferoxamine brain levels peaked between 100 and 200 mumol/L at 40 to 60 minutes after injection and exceeded serum levels by +/- 70%. Deferoxamine administered after induction of cerebral hypoxia-ischemia reduces injury in 7-day-old rats. Deferoxamine concentrates in the brain at levels between 100 and 200 mumol/L. At the concentrations achieved, deferoxamine might protect the brain through mechanisms unrelated to its ability to chelate iron.


[Emission computed tomography--new radiodiagnostic method].
A total of 224 patients with different diseases of the brain, lungs, heart, focal and chronic liver diseases, voluminous kidney changes and metastatic bone lesions were examined. A new radiodiagnostic method of emission computerized tomography was used in correlation with planar scintigraphy and x-ray computerized tomography. The author showed the value of the method in the diagnosis of the disturbed blood circulation of the organs. Its potentialities in the diagnosis of focal lesions were considered. A high informative value of the method as compared to statistical scintigraphy was noted.


Coamplification of multiple regions of chromosome 2, including MYCN, in a single patchwork amplicon in cancer cell lines.
Coamplification of multiple segments of chromosome 2, including an MYCN-bearing segment, was examined in 2 cancer cell lines, NCI-H69 (lung cancer) and IMR-32 (neuroblastoma). High-resolution array-CGH analysis revealed 13 and 6 highly amplified segments located at different sites in chromosome 2 in NCI-H69 and IMR-32, respectively. FISH analysis demonstrated that these segments were co-localized in double minutes in NCI-H69 and in homogeneously staining regions in IMR-32. Connectivity of the segments was determined by a PCR assay using designed primer sets. It was found that all the segments were connected to each other irrespective of their order and orientation against the genome sequence, and a single chain-like cluster was configured in both cell lines. Such patchwork structures of the amplicons suggest the possibility that massive genomic rearrangements, explained by the single catastrophic event model, are involved in the formation of the amplicons, enabling the coamplification of different chromosomal regions including the MYCN locus. The model comprises massive fragmentation of chromosomes and random rejoining of the fragments.


Role of dopamine in the neurotoxic effects of methamphetamine.
Multiple administrations of high doses of methamphetamine to rats cause long-term depression of both dopamine and serotonin synthesis. Coadministration of the catecholamine synthesis inhibitor, alpha-methyl-p-tyrosine, antagonizes this effect of methamphetamine on both neurotransmitter systems. However, when catecholamine synthesis was maintained by the administration of L-dopa and the peripheral decarboxylase inhibitor R04-4602, alpha-methyl-p-tyrosine no longer prevented the effects of methamphetamine on either dopamine or serotonin synthesis. In addition, the administration of the specific dopamine uptake blocker, amfonelic acid, significantly attenuated the changes in the serotonin synthesizing enzyme, tryptophan hydroxylase, resulting from multiple high doses of methamphetamine. The ability of a single administration of methamphetamine to depress tryptophan hydroxylase was also dependent on catecholamine synthesis. These results suggest that dopamine plays an important role in the changes mediated by the administration of methamphetamine in both the dopaminergic and serotonergic systems.