Comparison of induced versus natural regulatory T cells of the same TCR specificity for induction of tolerance to an environmental antigen.
Recent evidence shows that natural CD25(+)Foxp3(+) regulatory T cells (nTreg) and induced CD25(+)Foxp3(+) regulatory T cells (iTreg) both contribute to tolerance in mouse models of colitis and asthma, but there is little evidence regarding their relative contributions to this tolerance. We compared the abilities of nTreg and iTreg, both from OVA-TCR-transgenic OTII mice, to mediate tolerance in OVA-asthmatic C57BL/6 mice. The iTreg were differentiated from Th2 effector T cells by exposure to IL-10-differentiated dendritic cells (DC10) in vitro or in vivo, whereas we purified nTreg from allergen-naive mice and exposed them to DC10 before use. Each Treg population was subsequently repurified and tested for its therapeutic efficacy in vitro and in vivo. DC10 engaged the nTreg in a cognate fashion in Forster (or fluorescence) resonance energy transfer assays, and these nTreg reduced in vitro OVA-asthmatic Th2 effector T cell responses by 41-56%, whereas the comparator iTreg reduced these responses by 72-86%. Neutralization of IL-10, but not TGF-β, eliminated the suppressive activities of iTreg but not nTreg. Delivery of 5 × 10(5) purified nTreg reduced allergen challenge-induced airway IL-4 (p ≤ 0.03) and IL-5 (p ≤ 0.001) responses of asthmatic recipients by ≤ 23% but did not affect airway hyperresponsiveness or IgE levels, whereas equal numbers of iTreg of identical TCR specificity reduced all airway responses to allergen challenge by 82-96% (p ≤ 0.001) and fully normalized airway hyperresponsiveness. These data confirm that allergen-specific iTreg and nTreg have active roles in asthma tolerance and that iTreg are substantially more tolerogenic in this setting.


Enriched environment increases spinophilin mRNA expression and spinophilin immunoreactive dendritic spines in hippocampus and cortex.
Housing rodents in an enriched environment (EE) induces structural and functional plasticity in the adult brain, including increased dendritic sprouting and number of dendritic spines. However, the molecular mechanisms behind EE-induced brain plasticity remain largely unknown. Circadian rhythm plays an important role in memory processing but the neurobiological mechanisms of how circadian rhythm affects memory and brain plasticity remain controversial. In the current study, we studied the expression of spinophilin, a protein highly enriched in dendritic spines and involved in spine morphology and synaptic plasticity, to examine the effects of EE and circadian rhythm in rats housed in EE for different periods of time. Spinophilin mRNA expression was studied by in situ hybridization and the density of spinophilin immunoreactive puncta was quantified after immunohistochemical staining. Compared to rats living in a deprived environment (DE), we found a transient increase in the density of spinophilin immunoreactive puncta in hippocampus and cortex after 1 week of EE housing and persistent elevations of spinophilin mRNA expression during 1-4 weeks of environmental enrichment. Increased spinophilin expression was found during the light phase of the diurnal cycle, but not the dark phase. Thus, enriched housing altered the diurnal variation in spinophilin mRNA expression, suggesting that circadian modulation is likely to be important for experience dependent plasticity. The current results suggest a possible role for spinophilin in neuronal plasticity induced by environmental enrichment, but further studies are needed to establish a cause-effect relation.


Attenuated age-related thinning of peripapillary retinal nerve fiber layer in long eyes.
To assess the impact of axial length on the age-related peripapillary retinal nerve fiber layer (RNFL) thinning. This cross-sectional observational comparative case series included 172 eyes from 172 healthy Korean subjects. Peripapillary RNFL thickness was measured using an Optic Disc Cube 200 × 200 scan of spectral domain Cirrus HD OCT and the axial length was measured using IOL Master Advanced Technology. In age groups based on decade, the normal ranges of peripapillary RNFL thickness for average, quadrant, and clock-hour sectors were determined with 95% confidence intervals. After dividing the eyes into two groups according to axial length (cut-off, 24.50 mm), the degrees of age-related RNFL thinning were compared. Among the eyes included in the study, 53 (30.81%) were considered to be long eyes (axial length, 25.04 ± 0.48 µm) and 119 (69.19%) were short-to-normal length eyes (axial length, 23.57 ± 0.60 µm). The decrease in average RNFL thickness with age was less in long eyes (negative slope, -0.12 µm/yr) than in short-to-normal length eyes (negative slope, -0.32 µm/yr) (p < 0.001). Age-related thinning of peripapillary RNFL thickness is attenuated in long eyes compared to short-to-normal length eyes.


Trophic transfer patterns of arsenic in freshwater ecosystem layers in arsenic-endemic Ganges Delta and its potential human health risk.
Arsenic (As) is a toxic environmental contaminant with global public health concern. In aquatic ecosystems, the quantification of total As is restricted chiefly to the individual organisms. The present study has quantified the total As in different trophic layers (sediment-water-phytoplankton-periphyton-zooplankton-fish-gastropod-hydrophytes) of lentic freshwater ecosystems. As transfer pathways quantifying the transmission rate across trophic-level compartmental route were delineated using a novel model-based approach along with its potential contamination risk to humans. Lentic water bodies from Indo-Gangetic region, a core area of groundwater As, were selected for the present investigation. The study revealed that among the lower biota, zooplankton were the highest accumulator of total As (5554-11,564 µg kg-1) with magnification (rate = 1.129) of the metalloid, followed by phytoplankton (2579-6865 µg kg-1) and periphytic biofilm (1075 to 4382 µg kg -1). Muscle tissue of zooplanktivore Labeo catla is found to store higher As (80-115 µg kg-1 w.w.) compared to bottom-dwelling omnivore Cirrhinus mrigala (58-92 µg kg-1 w.w.). Whereas, Amblypharyngodon mola has accumulated higher As (203-319 µg kg-1 w.w.) than Puntius sophore (30-98 µg kg-1 w.w.) that raised further concern. The hepatic concentration indicated arsenic-mediated stress based on As stress index (threshold value = 1). Mrigal and Mola showed significant biomagnification among fishes while biodiminution was observed in Catla, Bata, Rohu and Punti. All the studied fishes were under the arsenic mediated stress. In the 'sediment-water-periphytic biofilm-gastropod' compartment, the direct grazing accumulation was higher (rate = 0.618) than the indirect path (rate = 0.587). Stems of edible freshwater macrophytes accumulated lesser As (32-190 µg kg-1 d.w.) than roots (292-946 µg kg-1 d.w.) and leaves (62-231 µg kg-1 d.w.). The target cancer risk (TCR) revealed a greater concern for adults consuming edible macrophyte regularly. Similarly, the varied level of target hazard quotient and TCR for adults consuming fishes from these waterbodies further speculated significant health concerns. The trophic transfer rate of environmental As in soil-water-biota level at an increasing trophic guild and consumer risk analysis have been unravelled for the first time in the Indo-Gangetic plains, which will be helpful for the strategic mitigation of As contamination.


Chronic cola drinking induces metabolic and cardiac alterations in rats.
To investigate the effects of chronic drinking of cola beverages on metabolic and echocardiographic parameters in rats. Forty-eight male Wistar rats were divided in 3 groups and allowed to drink regular cola (C), diet cola (L), or tap water (W) ad libitum during 6 mo. After this period, 50% of the animals in each group were euthanized. The remaining rats drank tap water ad libitum for an additional 6 mo and were then sacrificed. Rat weight, food, and beverage consumption were measured regularly. Biochemical, echocardiographic and systolic blood pressure data were obtained at baseline, and at 6 mo (treatment) and 12 mo (washout). A complete histopathology study was performed after sacrifice. After 6 mo, C rats had increased body weight (+7%, P < 0.01), increased liquid consumption (+69%, P < 0.001), and decreased food intake (-31%, P < 0.001). C rats showed mild hyperglycemia and hypertriglyceridemia. Normoglycemia (+69%, P < 0.01) and sustained hypertriglyceridemia (+69%, P < 0.01) were observed in C after washout. Both cola beverages induced an increase in left ventricular diastolic diameter (C: +9%, L: +7%, P < 0.05 vs W) and volumes (diastolic C: +26%, L: +22%, P < 0.01 vs W; systolic C: +24%, L: +24%, P < 0.05 vs W) and reduction of relative posterior wall thickness (C: -8%, L: -10%, P < 0.05 vs W). Cardiac output tended to increase (C: +25%, P < 0.05 vs W; L: +17%, not significant vs W). Heart rate was not affected. Pathology findings were scarce, related to aging rather than treatment. This experimental model may prove useful to investigate the consequences of high consumption of soft drinks.


Improved accuracy and speed in scanning probe microscopy by image reconstruction from non-gridded position sensor data.
Scanning probe microscopy (SPM) has facilitated many scientific discoveries utilizing its strengths of spatial resolution, non-destructive characterization and realistic in situ environments. However, accurate spatial data are required for quantitative applications but this is challenging for SPM especially when imaging at higher frame rates. We present a new operation mode for scanning probe microscopy that uses advanced image processing techniques to render accurate images based on position sensor data. This technique, which we call sensor inpainting, frees the scanner to no longer be at a specific location at a given time. This drastically reduces the engineering effort of position control and enables the use of scan waveforms that are better suited for the high inertia nanopositioners of SPM. While in raster scanning, typically only trace or retrace images are used for display, in Archimedean spiral scans 100% of the data can be displayed and at least a two-fold increase in temporal or spatial resolution is achieved. In the new mode, the grid size of the final generated image is an independent variable. Inpainting to a few times more pixels than the samples creates images that more accurately represent the ground truth.


Mechanism of carbon tetrachloride reduction in ferrous ion activated calcium peroxide system in the presence of methanol.
This study investigated the reductive initiation for the depletion of highly oxidized/perhalogenated pollutants, specifically the degradation of carbon tetrachloride (CT) was induced by adding methanol (MeOH) into a ferrous ion (Fe(II)) activated calcium peroxide (CaO2) system. The results indicated that CT could be completely degraded within 20 min at CaO2/Fe(II)/MeOH/CT molar ratio of 30/40/10/1 in this system. Scavenging tests suggested that both superoxide radical anion (O2 •-) and carbon dioxide radical anion (CO2 •-) were predominant reactive species responsible for CT destruction. Hydroxymethyl radicals (•CH2OH), an intermediate in the transformation of MeOH, could also initiate CT degradation by reducing C-Cl bond. GC/MS analysis identified CHCl3, C2Cl4, and C2Cl6 as the intermediates accompanied by CT destruction, and a reduction mechanism for CT degradation was proposed accordingly. In addition, the impact of solution matrix and initial solution pH were evaluated, and the results showed that Cl-, NO3 -, and HCO3 - had adverse effects on CT degradation. Moreover, the alkaline condition was unfavorable to CT depletion. In conclusion, the results obtained in the actual groundwater tests encouragingly demonstrated that the CaO2/Fe(II)/MeOH process is a highly promising technique for the remediation of CT-contaminated groundwater.


Operant behavior of the rat can be controlled by the configuration of objects in an animated scene displayed on a computer screen.
We developed a novel behavioral task in which rats learn to recognize the configuration of objects in an animated scene displayed on a computer screen. The scene consisted of a moving bar and a stationary rectangle. Rats deprived of food were trained to press a lever for reward in a small chamber located in front of the screen. Lever presses were rewarded only when the bar was at the rectangle. Rats anticipated the reward by gradually increasing frequency of lever pressing as the bar approached the rectangle. Control experiments showed that neither the timing nor the discrimination of rewarded and non-rewarded periods as two discrete conditions explain behavior of the rat. Because the changes in the scene were generated by movement of the object, the presented task could be used for studying neural structures involved in spatial behavior of rats using virtual reality technology.


Evaluation of protective effect of propolis on parotid salivary glands in gamma-irradiated rats.
One of the most significant side effects of radiotherapy for head and neck cancers is xerostomia as a result of salivary gland damage. Considering pharmaco- logical effects of propolis, we evaluated its protective effect on salivary glands subjected to radiotherapy of head and neck cancer patients. Twenty-one male albino rats (8-11 W, 190 ± 5 gm) were divided into three groups of seven animals. Scintigraphy was performed in all the groups. Then groups 1 (S) and 2 (SR) received normal saline injections and group 3 (PR) received propolis injection over 3 days. After that groups 2 and 3 were exposed to gamma radiation and all the rats underwent scintigraphic assessment on third day and 70th day after irradiation. The lips and tongues of rats in groups 2 and 3 were examined for mucositis daily in first 10 days. At the end, the parotid glands of all rats were examined histologically. Scintigraphy results of third and 70th day after irradiation showed statistically significant differences between PR and SR as well as SR and S. However, there was no significant difference between the PR and S groups. Histopathologic assessment demonstrated significant difference between SR, PR and S. These results suggest that propolis has protective effects on salivary gland function in animal models whilst it did not prevent radiation-induced histologic changes in tissues. Further investigations are needed to elucidate mechanisms of propolis actions. Clinical significance: Regarding to the results of this study, propolis may be useful in reduction xerostomia due to radiation to salivary glands and may be helpful for head and neck cancer patients.


Osteogenic induction of human periodontal ligament fibroblasts under two- and three-dimensional culture conditions.
Human periodontal ligament fibroblasts (hPDLF) play a key role in the regeneration of periodontal compartment during guided tissue regeneration procedures. This property is attributed to the progenitor cell subsets residing in the area. The aim of this study was to investigate whether hPDLFs could undergo an osteogenic differentiation under two- and three-dimensional (2D and 3D) culture conditions upon osteogenic induction. hPDLFs were isolated from six healthy donors, cultured, and expanded according to standard protocols. Then, three osteogenic culture conditions (dexamethasone, ascorbic acid, and beta-glycerophosphate) were established: 1) 2D culture as single-cell monolayer, 2) 3D-static culture on mineralized poly(DL-lactic-co-glycolic acid) (PLGA) scaffold, and 3) 3D culture on mineralized PLGA scaffold inside the NASA-approved bioreactor stimulating microgravity conditions. After 21 days of osteogenic induction, the majority of monolayer cultures had undergone differentiation toward osteogenic lineage, as indicated by morphological changes, mineralization assay, and some phenotypical properties. However, immunohistochemistry revealed that the scaffold cultures expressed higher levels of osteogenic marker proteins compared with that of the monolayers. Secondly, hPDLF-PLGA constructs in bioreactor showed an increased expression of osteopontin and osteocalcin compared with that of static 3D culture after 21 days. Results indicate that human periodontal ligament contains a subpopulation of cells capable of undergoing osteogenic differentiation and presumably contributing to regeneration of bone defects in the adjacent area. Human PDLF-seeded mineralized PLGA scaffold in microgravity bioreactor may be used to support osteogenic differentiation in vitro. Thus, this system may offer new potential benefits as a tool for periodontal tissue engineering.


Comparison of the transradial and transfemoral approaches for coronary angiographic evaluation in patients with internal mammary artery grafts.
Transradial access for coronary angiography may offer some advantages in comparison with conventional transfemoral access in specific subgroups of patients. Therefore, the aim of this study was to compare the transradial and transfemoral approaches in patients with previous coronary artery bypass graft (CABG) surgery using the left internal mammary artery (IMA). The time to left IMA cannulation, the time needed for left IMA evaluation, and total procedure time were compared in 60 patients previously submitted to CABG surgery using the left IMA. The homolateral radial access was attempted in 20 consecutive patients (TR group), whereas the transfemoral approach was undertaken in 40 patients (2: 1, TF group), matched for age, sex and number of grafts. In the TR group, the time to left IMA cannulation and the time needed for left IMA evaluation were 39% and 46%, respectively, lower than those in the TF group (P = 0.01 and P = 0.001, respectively), whereas total procedure time was similar. In the subgroup of patients with less than three saphenous vein grafts (n = 36), the time to left IMA cannulation, the time needed for left IMA evaluation as well as total procedure time were significantly reduced (P = 0.01, P = 0.001 and P = 0.004, respectively). The rate of ad-hoc coronary angioplasty procedures was unaffected by the access site (20% in the TR group and 27% in the TF group, P = 0.53). The present study shows that the homolateral transradial approach facilitates left IMA evaluation in patients with previous CABG surgery undergoing coronary angiography, in particular in patients with a lower number of coronary artery bypass conduits, in the absence of an excess of periprocedural complications.


Charge delocalization characteristics of regioregular high mobility polymers.
Controlling the regioregularity among the structural units of narrow bandgap conjugated polymer backbones has led to improvements in optoelectronic properties, for example in the mobilities observed in field effect transistor devices. To investigate how the regioregularity affects quantities relevant to hole transport, regioregular and regiorandom oligomers representative of polymeric structures were studied using density functional theory. Several structural and electronic characteristics of the oligomers were compared, including chain planarity, cation spin density, excess charges on molecular units and internal reorganizational energy. The main difference between the regioregular and regiorandom oligomers is found to be the conjugated backbone planarity, while the reorganizational energies calculated are quite similar across the molecular family. This work constitutes the first step on understanding the complex interplay of atomistic changes and an oligomer backbone structure toward modeling the charge transport properties.


The Gut-Lung Axis in Health and Respiratory Diseases: A Place for Inter-Organ and Inter-Kingdom Crosstalks.
The gut and lungs are anatomically distinct, but potential anatomic communications and complex pathways involving their respective microbiota have reinforced the existence of a gut-lung axis (GLA). Compared to the better-studied gut microbiota, the lung microbiota, only considered in recent years, represents a more discreet part of the whole microbiota associated to human hosts. While the vast majority of studies focused on the bacterial component of the microbiota in healthy and pathological conditions, recent works have highlighted the contribution of fungal and viral kingdoms at both digestive and respiratory levels. Moreover, growing evidence indicates the key role of inter-kingdom crosstalks in maintaining host homeostasis and in disease evolution. In fact, the recently emerged GLA concept involves host-microbe as well as microbe-microbe interactions, based both on localized and long-reaching effects. GLA can shape immune responses and interfere with the course of respiratory diseases. In this review, we aim to analyze how the lung and gut microbiota influence each other and may impact on respiratory diseases. Due to the limited knowledge on the human virobiota, we focused on gut and lung bacteriobiota and mycobiota, with a specific attention on inter-kingdom microbial crosstalks which are able to shape local or long-reached host responses within the GLA.


Effects of extremely low-frequency electromagnetic fields on delayed chromosomal instability induced by bleomycin in normal human fibroblast cells.
This study was carried out to examine the interaction of extremely low-frequency electromagnetic fields (ELF-EMF) on delayed chromosomal instability by bleomycin (BLM) in human fibroblast cells. A micronucleus-centromere assay using DNA probes for chromosomes 1 and 4 was performed and a 60-Hz ELF-EMF of 0.8 mT field strength was applied either alone or with BLM throughout the culture period. The frequencies of micronuclei (MN) and aneuploidy were analyzed at 28, 88, and 240 h after treatment with BLM. The coexposure of cells to BLM and ELF-EMF led to a significant increase in the frequencies of MN and aneuploidy compared to the cells treated with BLM alone. No difference was observed between field-exposed and sham-exposed control cells. The frequency of MN induced by BLM was increased at 28 h, and further analysis showed a persistent increase up to 240 h, but the new levels were not significantly different from the level at 28 h. BLM increased the frequencies of aneuploidy at 28, 88, and 240 h, and significantly higher frequency of aneuploidy was observed in the cells analyzed at 240 h compared to the cells examined at 28 h. No interaction of ELF-EMF on delayed chromosomal instability by BLM was observed. Our results suggest that ELF-EMF enhances the cytotoxicity of BLM. BLM might induce delayed chromosomal instability, but no effect of ELF-EMF was observed on the BLM-induced delayed chromosomal instability in fibroblast cells.


Targeting of glioma stem-like cells with a parthenolide derivative ACT001 through inhibition of AEBP1/PI3K/AKT signaling.
Glioblastoma (GBM) is the most lethal primary brain tumor in adults with a median survival of around 15 months. A potential treatment strategy involves targeting glioma stem-like cells (GSCs) that are able to initiate, maintain, and repopulate the tumor mass. Here, we identify ACT001, a parthenolide derivative, targeting GSCs through regulation of adipocyte enhancer binding protein 1 (AEBP1) signaling. Methods: The effects of ACT001 on cell survival of normal human astrocytes (NHA) and patient-derived glioma stem-like cells (GSCs) were evaluated. RNA-Seq were performed to detect differentially expressed genes. ACT001 efficacy as a single agent or in combination with SHP-2 inhibitor SHP099 was assessed using a GSC orthotopic xenograft model. Results: GSCs exhibit high response to ACT001 in compared with normal human astrocytes. AEBP1 is a putative target of ACT001 by RNA-Seq analysis, which expression associates with prognosis of GBM patients. Knockdown of AEBP1 inhibits GSC proliferation and glioma sphere formation. Treatment with ACT001 or PI3K inhibitor or AEBP1 depletion would impair AKT phosphorylation and GSC proliferation, whereas constitutive AKT activation rescues ACT001 treatment or AEBP1 depletion-inhibited cell proliferation. Moreover, ACT001 blocks TGF-β-activated AEBP1/AKT signaling in GSCs. ACT001 exhibits antitumor activity either as a single agent or in combination with SHP099, which provides significant survival benefits for GSC tumor xenograft-bearing animals. Conclusions: Our data demonstrate AEBP1 as a new druggable target in GBM and ACT001 as a potential therapeutic option for improving the clinical treatment of GBM in combination with SHP099.


The interface between glial progenitors and gliomas.
The mammalian brain and spinal cord contain heterogeneous populations of cycling, immature cells. These include cells with stem cell-like properties as well as progenitors in various stages of early glial differentiation. This latter population is distributed widely throughout gray and white matter and numerically represents an extremely large cell pool. In this review, we discuss the possibility that the glial progenitors that populate the adult CNS are one source of gliomas. Indeed, the marker phenotypes, morphologies, and migratory properties of cells in gliomas strongly resemble glial progenitors in many ways. We review briefly some salient features of normal glial development and then examine the similarities and differences between normal progenitors and cells in gliomas, focusing on the phenotypic plasticity of glial progenitors and the responses to growth factors in promoting proliferation and migration of normal and glioma cells, and discussing known mutational changes in gliomas in the context of how these might affect the proliferative and migratory behaviors of progenitors. Finally, we will discuss the "cancer stem cell" hypothesis in light of the possibility that glial progenitors can generate gliomas.


Adverse adolescent relationship histories and young adult health: cumulative effects of loneliness, low parental support, relationship instability, intimate partner violence, and loss.
To examine the associations between adverse interpersonal relationship histories experienced during adolescence and health in young adulthood in a large, nationally representative sample. Using data from Waves I, II, and III of the National Longitudinal Study of Adolescent Health, multiple adverse relationship experiences were examined, including high loneliness, low perceived parental support, frequent transitions in romantic relationships (relationship instability), exposure to intimate partner violence, and loss by death of important relationship figures. These histories are assessed, both individually and in a relationship risk index, as predictors of self-reported general health and depressive symptoms at Wave III (ages, 18-27), controlling for baseline (Wave I) health and for demographic and health behavior covariates. Net of baseline health and covariates, each type of relationship risk (experienced between Wave I and Wave III) was related to either depression or general health at Wave III, with the strongest effects seen for exposure to intimate partner violence. In addition, a cumulative relationship risk index examining the extent to which youth experienced high levels of multiple relationship risk factors revealed that each additional adverse relationship experience increased the odds of reporting worse mental and general health at Wave III, with increases occurring in an additive manner. Multiple types of adverse relationship experiences predicted increases in poorer general health and depressive symptoms from adolescence to early adulthood. Consistent with a cumulative risk hypothesis, the more types of adverse relationship a youth experienced, the worse were their young adult health outcomes.


Prevention of fractures in older people who fall.
Despite assessment and appropriate interventions, some older people will continue to be at risk of falling. Fractures of the hip and wrist are especially common among this group. The risk of fracture means that a comprehensive falls prevention programme must include an assessment of the individual fracture risk and the instigation of appropriate treatment of bone fragility. Osteoporosis is the main cause of bone fragility in this client group and it is commonly associated with postmenopausal women, although other forms exist. Osteoporosis is also a problem for men as they get older and for individuals taking steroid medications. Community nurses need to be aware of how osteoporosis manifests itself as well as how this condition can be prevented and treated.


Pheochromocytoma of the urinary bladder: a rare cause of severe hypertension.
Paraganglioma of the urinary bladder is a rare entity, and accounts for less than 0.5% of all bladder tumors. In the genitourinary tract, the urinary bladder is the most common site for paragangliomas. These tumors commonly present with hematuria and intermittent hyper-tension during micturition along with generalized symptoms due to raised catecholamines. Surgical removal is the treatment of choice, with most of the patients requiring at least a partial cystectomy. We herewith report a 38-year-old lady who presented with a two month history of pain abdomen, burning micturition, headache and uncontrolled hypertension. Detailed investigations revealed a pheochromocytoma of the urinary bladder, which was treated with partial cystectomy, following which her blood pressure stabilized.


Parents' Experiences of Information and Decision Making in the Care of Their Child With Severe Spinal Muscular Atrophy: A Population Survey.
This study aims to assess the experiences and wishes of parents of children with severe spinal muscular atrophy regarding information and decision-making throughout the course of the illness. A full population survey, conducted in 2015, among parents of children with severe spinal muscular atrophy who were born in Denmark between January 1, 2003, and December 31, 2013. We used a study-specific questionnaire with items about experiences and wishes concerning the provision of information about diagnosis, treatment, and end-of-life care. Among the 47 parents that were identified, 34 parents of 21 children participated. Eleven of them were nonbereaved and 23 were bereaved parents. All parents stated that health care staff did not take any decisions without informing them. A proportion of parents indicated that they were not informed about what spinal muscular atrophy entails (32%), possible treatment options (18%), or the fact that their child would have a short life (26%) or that death was imminent (57%). Most of the bereaved parents who had wishes concerning how and where their child would pass away had their wishes fulfilled. The study showed that health care staff did not take treatment decisions without parents being informed. However, there is room for improvement concerning information about what spinal muscular atrophy entails, treatment options, and prognosis. Possibilities of palliative care and advance care planning should be investigated for these parents, their child, and health care staff.


Drugs and punished responding. V. Effects of drugs on responding suppressed by response-dependent and response-independent electric shock.
A procedure was used where responses by a pigeon produced electric shock both to that pigeon (response-dependent shock) and to another pigeon (response-independent shock) working under the same schedule of food presentation. Both response-dependent and response-independent shock partially suppressed responding. Pentobarbital, chlordiazepoxide and ethanol increased responding suppressed by response-dependent shock, but only pentobarbital increased responding suppressed by response-independent shock. Morphine, d-amphetamine, chlorpromazine, doxepin, and pentazocine either had no effect, or further decreased rates of responding suppressed by response-dependent or response-independent shock.


Transfer of 360° virtual reality and match broadcast video-based tests to on-field decision-making.
This study aimed to assess the level of transfer of two reliable and valid video modes to in-game decision-making performance. Two video-based tests of 25 clips each (360°VR and match broadcast vision) assessed off-field decision-making accuracy in elite Australian football umpires (n=21). Game performance was assessed across four games for each participant, classified into two groups based on this measure; "highly skilled" or 'skilled'. Decision-making was assessed for correct, missed and unwarranted decisions in video-based tests and in-game assessments. Independent t-tests analysed differences between highly skilled and skilled in-game decision-makers for each test. Correlations also compared experience and in-game with video-based test decision-making performance. For both video-based tests, there were no significant differences between highly skilled and skilled in-game decision-makers, nor any significant correlations. Officials who made less unwarranted decisions in-game (highly skilled) made significantly less unwarranted decisions in the match broadcast test. There was a significant correlation between experience and 360°VR correct decision-making. Neither video-based test had the sensitivity to discriminate between elite officials, potentially due to the third-person perspective (match broadcast task) or sub-elite players presented (360°VR). Optimising the representativeness of off-field tasks through including similar constraints to performance environments is an important consideration for researchers and practitioners.


Nanoscopic surfactant behavior of the porin MspA in aqueous media.
The mycobacterial porin MspA is one of the most stable channel proteins known to date. MspA forms vesicles at low concentrations in aqueous buffers. Evidence from dynamic light scattering, transmission electron microscopy and zeta-potential measurements by electrophoretic light scattering indicate that MspA behaves like a nanoscale surfactant. The extreme thermostability of MspA allows these investigations to be carried out at temperatures as high as 343 K, at which most other proteins would quickly denature. The principles of vesicle formation of MspA as a function of temperature and the underlying thermodynamic factors are discussed here. The results obtained provide crucial evidence in support of the hypothesis that, during vesicle formation, nanoscopic surfactant molecules, such as MspA, deviate from the principles underlined in classical surface chemistry.


Genetically encodable bioluminescent system from fungi.
Bioluminescence is found across the entire tree of life, conferring a spectacular set of visually oriented functions from attracting mates to scaring off predators. Half a dozen different luciferins, molecules that emit light when enzymatically oxidized, are known. However, just one biochemical pathway for luciferin biosynthesis has been described in full, which is found only in bacteria. Here, we report identification of the fungal luciferase and three other key enzymes that together form the biosynthetic cycle of the fungal luciferin from caffeic acid, a simple and widespread metabolite. Introduction of the identified genes into the genome of the yeast Pichia pastoris along with caffeic acid biosynthesis genes resulted in a strain that is autoluminescent in standard media. We analyzed evolution of the enzymes of the luciferin biosynthesis cycle and found that fungal bioluminescence emerged through a series of events that included two independent gene duplications. The retention of the duplicated enzymes of the luciferin pathway in nonluminescent fungi shows that the gene duplication was followed by functional sequence divergence of enzymes of at least one gene in the biosynthetic pathway and suggests that the evolution of fungal bioluminescence proceeded through several closely related stepping stone nonluminescent biochemical reactions with adaptive roles. The availability of a complete eukaryotic luciferin biosynthesis pathway provides several applications in biomedicine and bioengineering.


Addiction and the Dark Triad of Personality.
In this article, we review associations between the Dark Triad of personality (narcissism, Machiavellianism, and psychopathy) and addictive behaviors, both substance-related and non-substance-related. We summarize evidence from personality and clinical research and integrate it with prevailing models of addiction. Specifically, we discuss addictive behavior in the light of affect regulation, which is likely more relevant in narcissism, as well as inhibitory deficits, a putative mechanism in psychopathy. These mechanisms can be related to central motives of the respective personality constructs, such as stabilization of self-esteem in narcissism and impulsive stimulation seeking in psychopathy. We conclude that different mechanisms might lead to similar observable behavior in narcissism and psychopathy at earlier stages of the addiction cycle, but psychopathic disinhibition might be particularly relevant at later stages. This underpins the importance of considering personality factors for the understanding and treatment of addiction.


Diode-Pumped Nd:FAP Laser at 1.126 microm: A Possible Local Oscillator for a Hg+ Optical Frequency Standard.
We report the efficient operation of a continuous-wave, single-frequency, diode-pumped Nd:FAP laser at 1.126 mum. When frequency quadrupled, such a laser might be used as a local oscillator for an optical frequency standard based on the single-photon (2)S(1/2)-(2)D(5/2) electric quadrupole transition of a trapped and laser-cooled (199)Hg(+) ion. Since the frequencies of the atomic transition and the laser are harmonically related, this scheme helps to simplify the measurement of the S-D clock transition frequency by a phase-coherent chain to the cesium primary frequency standard.


A critical review of the quality and safety of BADGE-based epoxy coatings for cans: implications for legislation on epoxy coatings for food contact.
BADGE-based epoxy resins have been commercially available for more than 40 years. They are extremely versatile, finding applications in many fields in both thermal and ambient cure applications. The present review focuses on their use in the food industry as surface coatings for food and beverage cans, and in particular on their chemical stability in relation to their toxicology and sensorial effects on foods. The implications of these properties of epoxy resins for prospective EU legislation on plastic coatings for food contact is discussed.


Assessment of morphology for anaerobic-granular particles.
This study shows that a proper assessment of granule morphology is fundamental before applying any mathematical derivation or empirical formula based on the shape factor of anaerobic-granular particles to determine granular characteristics. The granular images and size distribution of samples observed in this study revealed two different dimensions along two axes, which characterize these particles as ellipsoids. In the literature, theoretical-settling velocities and particle-size distribution of anaerobic granules have been calculated by assuming granules as spherical-shaped and using the numerical correlation between the size and settling velocity. This resulted in large deviations in results reported for settling velocities and size distribution calculated by using empirical equations when compared with experimentally measured values. It is believed that, because of the nonspherical nature of these particles, errors have been made in the earlier studies (which considered these particles as spherical), while evaluating granular-physical properties.


Second heart field and the development of the outflow tract in human embryonic heart.
The second heart field (SHF) is indicated to contribute to the embryonic heart development. However, less knowledge is available about SHF development of human embryo due to the difficulty of collecting embryos. In this study, serial sections of human embryos from Carnegie stage 10 (CS10) to CS16 were stained with antibodies against Islet-1 (Isl-1), Nkx2.5, GATA4, myosin heavy chain (MHC) and α-smooth muscle actin (α-SMA) to observe spatiotemporal distribution of SHF and its contribution to the development of the arterial pole of cardiac tube. Our findings suggest that during CS10 to CS12, SHF of the human embryo is composed of the bilateral pharyngeal mesenchyme, the central mesenchyme of the branchial arch and splanchnic mesoderm of the pericardial cavity dorsal wall. With development, SHF translocates and consists of ventral pharyngeal mesenchyme and dorsal wall of the pericardial cavity. Hence, the SHF of human embryo shows a dynamic spatiotemporal distribution pattern. The formation of the Isl-1 positive condense cell prongs provides an explanation for the saddle structure formation at the distal pole of the outflow tract. In human embryo, the Isl-1 positive cells of SHF may contribute to the formation of myocardial outflow tract (OFT) and the septum during different development stages.


Nuclear quantum effects affect bond orientation of water at the water-vapor interface.
Using combined theoretical and experimental approaches, we demonstrate that the bond orientation of water at the water-vapor interface depends markedly on the water isotope (H-D) composition. While the interfacial water structures of H(2)O and D(2)O are indistinguishable, the intramolecular symmetry breaking in HDO is directly reflected at the surface: the OD bonds preferably orient down towards the bulk water, whereas the OH bond tends to orient up into the vapor phase. Path integral molecular dynamics simulations show good agreement with surface-specific sum-frequency generation (SFG) spectroscopy results, revealing that the distinct interfacial bond orientations originate from nuclear quantum effects. The enhanced localization of the heavier D atom leads to stronger hydrogen bonds, giving rise to OD bonds pointing down into the bulk.


How do culture media influence in vitro perivascular cell behavior?
Perivascular cells are multilineage cells located around the vessel wall and important for wall stabilization. In this study, we evaluated a stem cell media and a perivascular cell-specific media for the culture of primary perivascular cells regarding their cell morphology, doubling time, stem cell properties, and expression of cell type-specific markers. When the two cell culture media were compared to each other, perivascular cells cultured in the stem cell medium had a more elongated morphology and a faster doubling rate and cells cultured in the pericyte medium had a more typical morphology, with several filopodia, and a slower doubling rate. To evaluate stem cell properties, perivascular cells, CD146(-) cells, and mesenchymal stem cells (MSCs) were differentiated into the adipogenic, osteogenic, and chondrogenic lineages. It was seen that perivascular cells, as well as CD146(-) cells and MSCs, cultured in stem cell medium showed greater differentiation than cells cultured in pericyte-specific medium. The expression of pericyte-specific markers CD146, neural/glial antigen 2 (NG2), platelet-derived growth factor receptor-β (PDGFR-β), myosin, and α-smooth muscle actin (α-SMA) could be found in both pericyte cultures, as well as to varying amounts in CD146(-) cells, MSCs, and endothelial cells. The here presented work shows that perivascular cells can adapt to their in vitro environment and cell culture conditions influence cell functionality, such as doubling rate or differentiation behavior. Pericyte-specific markers were shown to be expressed also from cells other than perivascular cells. We can further conclude that CD146(+) perivascular cells are inhomogeneous cell population probably containing stem cell subpopulations, which are located perivascular around capillaries.


A Provably Convergent Control Closure Scheme for the Method of Moments of the Chemical Master Equation.
In this article, we introduce a novel moment closure scheme based on concepts from model predictive control (MPC) to accurately describe the time evolution of the statistical moments of the solution of the chemical master equation (CME). The method of moments, a set of ordinary differential equations frequently used to calculate the first nm moments, is generally not closed since lower-order moments depend on higher-order moments. To overcome this limitation, we interpret the moment equations as a nonlinear dynamical system, where the first nm moments serve as states, and the closing moments serve as the control input. We demonstrate the efficacy of our approach using three example systems and show that it outperforms existing closure schemes. For polynomial systems, which encompass all mass-action systems, we provide probability bounds for the error between true and estimated moment trajectories. We achieve this by combining the convergence properties of a priori moment estimates from stochastic simulations with guarantees for nonlinear reference tracking MPC. Our proposed method offers an effective solution to accurately predict the time evolution of moments of the CME, which has wide-ranging implications for many fields, including biology, chemistry, and engineering.


Specific MAIT cell behaviour among innate-like T lymphocytes in critically ill patients with severe infections.
In between innate and adaptive immunity, the recently identified innate-like mucosal-associated invariant T (MAIT) lymphocytes display specific reactivity to non-streptococcal bacteria. Whether they are involved in bacterial sepsis has not been investigated. We aimed to assess the number and the time course of circulating innate-like T lymphocytes (MAIT, NKT and γδ T cells) in critically ill septic and non-septic patients and to establish correlations with the further development of intensive care unit (ICU)-acquired infections. We prospectively enrolled consecutive patients with severe sepsis and septic shock. Controls were critically ill patients with non-septic shock and age-matched healthy subjects. Circulating innate-like lymphocytes were enumerated using a flow cytometry assay at day 1, 4 and 7. One hundred and fifty six patients (113 severe bacterial infections, 36 non-infected patients and 7 patients with severe viral infections) and 26 healthy subjects were enrolled into the study. Patients with severe bacterial infections displayed an early decrease in MAIT cell count [median 1.3/mm(3); interquartile range (0.4-3.2)] as compared to control healthy subjects [31.1/mm(3) (12.1-45.2)], but also to non-infected critically ill patients [4.3/mm(3) (1.4-13.2)] (P < 0.0001 for all comparisons). In contrast NKT and γδ T cell counts did not differ between patients groups. The multivariate analysis identified non-streptococcal bacterial infection as an independent determinant of decrease in MAIT cell count. Furthermore, the incidence of ICU-acquired infections was higher in patients with persistent MAIT cell depletion. This large human study provides valuable information about MAIT cells in severe bacterial infections. The persistent depletion of MAIT cells is associated with the further development of ICU-acquired infections.


Electronic structure optimizing of Ru nanoclusters via Co single atom and N, S co-doped reduced graphene oxide for accelerating water electrolysis.
The development of efficient and stable electrocatalysts for hydrogen evolution reaction (HER) is impending for the advancement of water-splitting. In this study, we developed a novel electrocatalyst consisting of highly dispersed Ru nanoclusters ameliorated by cobalt single atoms and N, S co-doped reduced graphene oxide (CoSARuNC@NSG). Benefitted from the optimized electronic structure of the Ru nanoclusters induced by the adjacent single atomic Co and N, S co-doped RGO support, the electrocatalyst exhibits exceptional HER performance with overpotentials of 15 mV and 74 mV for achieving a current density of 10 mA cm-2 in alkaline and acidic water. The catalyst outperforms most noble metal-based HER electrocatalysts. Furthermore, the electrolyzer assembled with CoSARuNC@NSG and RuO2 demonstrated an overall voltage of 1.56 V at 10 mA cm-2 and an excellent operational stability for over 25 h with almost no attenuation. Theoretical calculations also deduce its high HER activity demonstrated by the smaller reaction energy barrier due to the optimized electronic structure of Ru nanoclusters. This strategy involving the regulation of metal nanoparticles activity through flexible single atom and GO support could provide valuable insights into the design of high-performance and low-cost HER catalysts.


Tyrosine Nitration Contributes to Nitric Oxide-Stimulated Degradation of CYP2B6.
Human cytochrome P450 (P450) CYP2B6 undergoes nitric oxide (NO)-dependent proteasomal degradation in response to the NO donor dipropylenetriamine NONOate (DPTA) and biologic NO in HeLa and HuH7 cell lines. CYP2B6 is also downregulated by NO in primary human hepatocytes. We hypothesized that NO or derivative reactive nitrogen species may generate adducts of tyrosine and/or cysteine residues, causing CYP2B6 downregulation, and selected Tyr and Cys residues for mutation based on predicted solvent accessibility. CYP2B6V5-Y317A, -Y380A, and -Y190A mutant proteins expressed in HuH7 cells were less sensitive than wild-type (WT) enzyme to degradation evoked by DPTA, suggesting that these tyrosines are targets for NO-dependent downregulation. The Y317A or Y380A mutants did not show increases in high molecular mass (HMM) species after treatment with DPTA or bortezomib + DPTA, in contrast to the WT enzyme. Carbon monoxide-releasing molecule 2 treatment caused rapid suppression of 2B6 enzyme activity, significant HMM species generation, and ubiquitination of CYP2B6 protein but did not stimulate CYP2B6 degradation. The CYP2B6 inhibitor 4-(4-chlorophenyl)imidazole blocked NO-dependent CYP2B6 degradation, suggesting that NO access to the active site is important. Molecular dynamics simulations predicted that tyrosine nitrations of CYP2B6 would cause significant destabilizing perturbations of secondary structure and remove correlated motions likely required for enzyme function. We propose that cumulative nitrations of Y190, Y317, and Y380 by reactive nitrogen species cause destabilization of CYP2B6, which may act synergistically with heme nitrosylation to target the enzyme for degradation. SIGNIFICANCE STATEMENT: This work provides novel insight into the mechanisms by which nitric oxide, which is produced in hepatocytes in response to inflammation, triggers the ubiquitin-dependent proteasomal degradation of the cytochrome P450 (P450) enzyme CYP2B6. Our data demonstrate that both nitration of specific tyrosine residues and interaction of nitric oxide (NO) with the P450 heme are necessary for NO to trigger ubiquitination and protein degradation.


Determinants and Management of the Post-Pulmonary Embolism Syndrome.
Acute pulmonary embolism (PE) is not only a serious and potentially life-threatening disease in the acute phase, in recent years it has become evident that it may also have a major impact on a patient's daily life in the long run. Persistent dyspnea and impaired functional status are common, occurring in up to 50% of PE survivors, and have been termed the post-PE syndrome (PPES). Chronic thromboembolic pulmonary hypertension is the most feared cause of post-PE dyspnea. When pulmonary hypertension is ruled out, cardiopulmonary exercise testing can play a central role in investigating the potential causes of persistent symptoms, including chronic thromboembolic pulmonary disease or other cardiopulmonary conditions. Alternatively, it is important to realize that post-PE cardiac impairment or post-PE functional limitations, including deconditioning, are present in a large proportion of patients. Health-related quality of life is strongly influenced by PPES, which emphasizes the importance of persistent limitations after an episode of acute PE. In this review, physiological determinants and the diagnostic management of persistent dyspnea after acute PE are elucidated.


Classification of oolong tea varieties based on computer vision and convolutional neural networks.
In the contemporary food industry, accurate and rapid differentiation of oolong tea varieties holds paramount importance for traceability and quality control. However, achieving this remains a formidable challenge. This study addresses this lacuna by employing machine learning algorithms - namely support vector machines (SVMs) and convolutional neural networks (CNNs) - alongside computer vision techniques for the automated classification of oolong tea leaves based on visual attributes. An array of 13 distinct characteristics, encompassing color and texture, were identified from five unique oolong tea varieties. To fortify the robustness of the predictive models, data augmentation and image cropping methods were employed. A comparative analysis of SVM- and CNN-based models revealed that the ResNet50 model achieved a high Top-1 accuracy rate exceeding 93%. This robust performance substantiates the efficacy of the implemented methodology for rapid and precise oolong tea classification. The study elucidates that the integration of computer vision with machine learning algorithms constitutes a promising, non-invasive approach for the quick and accurate categorization of oolong tea varieties. The findings have significant ramifications for process monitoring, quality assurance, authenticity validation and adulteration detection within the tea industry. © 2023 Society of Chemical Industry.


Simultaneous bilateral acute angle-closure glaucoma in a patient with subarachnoid hemorrhage.
To present an unusual case of simultaneous bilateral acute angle-closure (AAC) glaucoma in a patient with subarachnoid hemorrhage due to an aneurysm involving the right middle cerebral artery. A 60-year-old woman with a clinically inexplicable bilateral nonreacting mydriasis after brain surgery underwent an ophthalmologic consultancy. The bilateral mydriasis was diagnosed as an unusual clinical presentation of simultaneous bilateral AAC glaucoma. One week after the AAC was bilaterally resolved, the patient underwent echographic examination because of the outbreak of a unilateral relapsed ocular hypertensive attack that required an ultrasound biomicroscopy (UBM) to be accurately diagnosed and treated. The bilateral AAC regressed completely after pharmacologic therapy. UBM evaluation of the eye with recurrence of the angle-closure glaucoma attack was necessary to confirm the diagnosis of unilateral relapsed angle-closure glaucoma due to an unresolved pupillary block. A viscoelastic-aided opening of the angle and peripheral surgical iridectomy were performed. UBM and tonometry were performed intraoperatively to confirm the success of the surgical treatment. The UBM allowed us to identify the pupillary block as the main mechanism involved in the pathogenesis of this unusual case of simultaneous bilateral AAC glaucoma. Despite the fact that drug-induced supraciliary uveal effusion and mydriasis due to surgical anesthesia have been frequently reported to contribute to this complication, we presumed that simultaneous intravenous administration of sedative drugs and adrenergic agonists, in a patient with individual biometric predisposing factors to the angle-closure, created the anatomic conditions which induced the pupillary block with obliteration of the trabeculum.


Skin-resident stem cells and wound healing.
CD271 is common stem cell marker for the epidermis and dermis. We assessed a kinetic movement of epidermal and dermal CD271+ cells in the wound healing process to elucidate the possible involvement with chronic skin ulcers. Epidermal CD271+ cells were proliferated and migrated from 3 days after wounding. Purified epidermal CD271+ cells expressed higher TGFβ2 and VEGFα transcripts than CD271- cells. Delayed wound healing was observed in the aged mice compared with young mice. During the wound healing process, the peak of dermal CD271+ cell accumulation was delayed in aged mice compared with young mice. The expression levels of collagen-1, -3, -5, F4-80, EGF, FGF2, TGFβ1, and IL-1α were significantly increased in young mice compared with aged mice. Furthermore, purified dermal CD271+ cells expressed higher FGF2, EGF, PDGFB, and TGFβ1 gene transcripts than CD271- cells. These results suggested that epidermal and dermal CD271+ cells were closely associated with wound healing process by producing various growth factors. Epidermal and dermal CD271+ cells in chronic skin ulcer patients were significantly reduced compared with healthy controls. Thus, both epidermal and dermal stem cells can play an important role in wound healing process.


Gender Identity, Ethnicity, Acculturation, and Drug Use: Exploring Differences among Adolescents in the Southwest.
This article presents the findings of a survey completed by 1351 predominantly Mexican American middle school students residing in a large urban center in the U.S. Southwest. The study explores possible associations between drug use attitudes and behaviors and gender (biological sex), gender identity, ethnicity, and acculturation status. Based on the concepts of "machismo" and "marianismo" that have been used to describe Mexican populations, four dimensions of gender identity were measured: aggressive masculinity, assertive masculinity, affective femininity, and submissive femininity. In explaining a variety of indicators of drug use behaviors and anti-drug norms, gender alone had limited explanatory power, while gender identity-often regardless of gender-was a better predictor. Aggressive masculinity was generally associated with higher risk of drug use, while the other three gender identity measures had selected protective effects. However, the impact of gender identity was strongly mediated by acculturation. Less acculturated Mexican American students reported lower aggressive masculinity scores than non-Latinos. Less acculturated Mexican American girls reported both the lowest aggressive masculinity scores and the highest submissive femininity scores. More acculturated Mexican American students, along with the less acculturated Mexican American boys, did not appear to be following a polarized approach to gender identity (machismo and marianismo) as was expected. The findings suggest that some aspects of culturally prescribed gender roles can have a protective effect against drug use behaviors and attitudes, possibly for both girls and boys.


Music Therapy in Hospice and Palliative Care: a Review of the Empirical Data.
Although music therapy is an established allied health profession and is used with increasing frequency in the treatment of those with a terminal illness, there is a real dearth of empirical research literature supporting the use of music therapy in end-of-life care. This article reviews the empirical studies found in the literature and documents the emergence of an evidenced-based approach to the use of music therapy in hospice and palliative care. A total of 11 studies are reviewed; of these, six show significant differences supporting the use of music therapy in this area. Dependent variables positively affected by music therapy include pain, physical comfort, fatigue and energy, anxiety and relaxation, time and duration of treatment, mood, spirituality and quality of life. Guidelines for future research are considered, and variables that need to be controlled are presented. The need to create an evidence-based approach to hospice and palliative care music therapy is articulated, and future researchers are empowered to continue to conduct investigations among this population.


Clonality, Heterogeneity, and Evolution of Synchronous Bilateral Ovarian Cancer.
Synchronous bilateral ovarian cancer (SBOC) represents a relatively frequent occurrence and clinically relevant diagnostic dilemma. Delineation of its clonal architecture, genetic heterogeneity, and evolutionary trajectories may have important implications for prognosis and management of patients with SBOC. Here, we describe the results of next-generation whole-exome or whole-genome sequencing of specimens from 12 SBOC cases and report that bilateral tumors from each individual display a comparable number of genomic abnormalities and similar mutational signatures of single-nucleotide variations. Clonality indices based on tumor-specific alterations supported monoclonal origins of SBOC. Each of the ovarian lesions was nevertheless oligoclonal, with inferred metastatic tumors harboring more subclones than their primary counterparts. The phylogenetic structure of SBOC indicated that most cancer cell dissemination occurred early, when the primary carcinoma was still relatively small (<100 million cells). Accordingly, the mutation spectra and mutational signatures of somatic variants exhibited pronounced spatiotemporal differences in each patient. Overall, these findings suggest that SBOCs are clonally related and form through pelvic spread rather than independent multifocal oncogenesis. Metastatic dissemination is often an early event, with dynamic mutational processes leading to divergent evolution and intratumor and intertumor heterogeneity, ultimately contributing substantially to phenotypic plasticity and diverse clinical course in SBOC. Cancer Res; 77(23); 6551-61. ©2017 AACR.


In vivo analysis of the Hsp90 cochaperone Sti1 (p60).
Hsp90 interacts with Sti1 (p60) in lysates of yeast and vertebrate cells. Here we provide the first analysis of their interaction in vivo. Saccharomyces cerevisiae mutations that eliminate Sti1 or reduce intracellular concentrations of Hsp90 individually have little or no effect on growth at normal temperatures. However, when combined, the mutations greatly reduce or eliminate growth. Furthermore, overexpression of Sti1 has allele-specific effects on cells carrying various hsp90ts point mutations. These genetic interactions provide strong evidence that Hsp90 and Sti1 interact in vivo and that their functions are closely allied. Indeed, deletion of STI1 reduces the in vivo activity of the Hsp90 target protein, glucocorticoid receptor (GR). Mutations in GR that eliminate interaction with Hsp90 also eliminate the effects of the sti1 deletion. Examination of GR protein complexes in the sti1 deletion mutant reveals a selective increase in the concentration of GR-Ydj1 complexes, supporting previous hypotheses that Ydj1 functions at an early step in the maturation of GR and that Sti1 acts at an intermediate step. Deletion of STI1 also reduces the in vivo activity of another, unrelated Hsp90 target protein, v-Src. Our data indicate that Sti1 is a general factor in the maturation of Hsp90 target proteins and support earlier suggestions that Hsp90 matures even very different target proteins by a similar mechanism.


Changes in the microvasculature of ischemic and infarcted myocardium.
The fine structure of the small vessels in experimental myocardial infarcts of 10- to 360-minute duration was studied in 32 dogs and compared with the appearance of the small vessels in the corresponding normal myocardium. Following 10 to 60 minutes of coronary artery occlusion, increasing numbers of endothelial cells showed a marked swelling which was consistent with the presence of an intracellular edema and frequently resulted in various degrees of obstruction of the vessel lumen. After 120 minutes of ischemia, not all endothelial cells were obviously swollen, but all showed signs of degeneration, including changes in organelles similar to those in surrounding muscle cells. A progressive intensification of degenerative changes, particularly with respect to the continuity of the endothelial lining and the integrity of the membrane systems of individual endothelial cells, was observed at 3, 4, 5, and 6 hours after coronary artery ligation, and cell debris could be seen in the lumina of most small vessels.


Use of white blood cell counts to predict metabolic syndrome in the elderly: a 4 year longitudinal study.
The white blood cell (WBC) count was one of the first inflammatory markers associated with metabolic syndrome (MetS). Recently, two longitudinal studies have demonstrated a cause and effect relationship between MetS and WBC counts among middle-aged adults. However, no study has used WBC cutoff values to predict MetS in the elderly. Subjects who underwent routine health checkups, and were above 60 years of age, were enrolled. All subjects were followed-up until they developed MetS or until 4 years from the date of entry, whichever came earlier. Of the 4539 subjects eligible for enrollment, 3428 subjects comprised the study group and 1111 subjects comprised the validation group. WBC counts were significantly different between subjects with and without MetS in both genders. Using the ROC curve, WBC cutoff values of 5.7 × 10(3)/µl in males and 5.0 × 10(3)/µl in females were associated with the increased risk of developing MetS (all p values <0.001). Using these WBC cutoff values, the hazard ratio (HR) for females was significant in both the study group and validation group. However, the HR for males failed significance in the validation group. Kaplan-Meier plots and κ coefficients confirmed that the WBC cutoff value could predict development of MetS in women but not in men. The association between WBC count and MetS was gender specific. A WBC cutoff value greater than 5.0 10(3)/µl may predict the development of MetS in elderly women.


Effect of gamma irradiation on soybean allergen levels.
The levels of food allergens in gamma-irradiated soybean (0, 2.5, 5, 7.5, 10, 20, and 30 kGy) were investigated by immunoblotting and ELISA, using allergen-specific antibodies and patient serum. After 3 months of storage, Coomassie brilliant blue (CBB) staining indicated similar total protein profiles among the treatments, but that some proteins were degraded by irradiation at high doses. Immunoblotting with specific antibodies for major soybean allergens (β-conglycinin, Gly m Bd 30 K, soybean trypsin inhibitor, and Gly m 4) resulted in apparent band profiles and intensities that were not significantly changed by irradiation. Competitive inhibition ELISA analyses suggested that there were no significant changes in the allergen contents, except for a decrease in the soybean trypsin inhibitor. The patient IgE binding allergenic protein patterns were not changed by irradiation up to 30 kGy. ELISA using patient serum also revealed that the IgE reactivity to the irradiated soybean extract did not increase from the level of the control, but that the reactivity to some patient serum IgE was significantly decreased by irradiation.


Surgery versus radioiodine therapy as definitive management for graves' disease: the role of patient preference.
Thyroidectomy is an option for the definitive management of Graves' disease. The aim of this study was to examine the role of patient preference for selecting surgery as definitive treatment. This is a retrospective cohort study comprising all patients (n = 63) presenting to a single surgeon for surgical management of Graves' disease over 3 years. Documented reasons for surgery were compared with accepted indications, as well as patients' perceptions as assessed by questionnaire. The most frequent absolute indication was the presence of a large goiter (n = 8; 13%) or associated thyroid nodule (n = 6; 10%). Ophthalmopathy, a relative indication, comprised the largest single group overall (n = 18; 29%); however, a significant number of patients (n = 17; 27%) elected surgery in the absence of a recognized indication. There was strong concordance (73%) between the recorded indication and the patients' survey response. Overall, there was a high level of satisfaction with surgery with 88% of respondents giving a satisfaction score of 7 or greater on a visual analog scale (VAS) (0-10). One-third of all patients electing surgery as definitive management do so in the absence of a specific indication. Overall, there is a high level of satisfaction with the decision for surgery as definitive management of Graves' disease.


Cochlear otosclerosis.
The aim of this study is to summarize current advances in research and clinical aspects of cochlear otosclerosis. Recent studies have revealed that otosclerosis is a process of bone remodeling that is unique to the otic capsule only. Even though no obvious bone remodeling is seen in the otic capsule under normal conditions, remodeling starts when some molecular factors trigger the capsule in certain patients who have genetic and/or environmental tendencies. Cochlear otosclerosis is defined as otosclerosis located in the otic capsule involving the cochlear endosteum and causing sensorineural hearing loss or mixed-type hearing loss. It has been clearly shown that, when otosclerosis is sufficiently severe to involve the cochlear endosteum, it usually fixes the stapes as well.


Oxidative stress after moderate to extensive burning in humans.
Lipid peroxidation products, lipid antioxidants, and hematologic and blood chemistry changes were evaluated in plasma of patients after acute burning injury involving 10% (n=8), 20% (n=8), and 40% (n=5) of total body surface area (TBSA), 24 h after burning (baseline) up to 30 days after. Markedly increased plasma levels of malondialdehyde (MDA) were observed at baseline in all patients, according to the extent of the injury, then the values declined progressively. However, levels of MDA remained above normal up to 30 days even in less injured patients. On the other hand, the plasma level of conjugated diene lipid hydroperoxides was only slightly higher than control at the baseline, then dropped under the control value in all patients. Cholesterol showed a marked fall at baseline, followed by a rapidly progressive decrease, indicating a massive loss of circulating lipids by the acute thermal injury. Because of such an extensive and rapidly spreading oxidative degradation of lipids, decomposition of conjugated diene hydroperoxides, produced in early stages of the peroxidation process, occurs, so these compounds cannot be a suitable index to value lipid oxidation in burned patients. Aldehydic products of lipid peroxidation act as endotoxins, causing damage to various tissues and organs. Damage to liver and decrease of erythrocyte survival were assessed by increased plasma levels of asparate and alanine transaminases, within 7-15 days after injury, and by a decreased number of red blood cells, which remained under the normal value at 30 days. A marked decrease of lipid antioxidants, beta-carotene, vitamin A and vitamin E was observed at baseline. The level of beta-carotene remained low in all patients at the end of the 30-day observation. A complete recovery of vitamin A did not occur at 30 days post-burn, even in the patients with 10% of burned TBSA. Plasma levels of vitamin E decreased significantly in 1-7 days after burn in all patients, but these levels increased thereafter, with almost total recovery at 30 days. These data show evidence of a marked, long-lasting oxidant/antioxidant imbalance in burned patients, in accordance with the severity of the injury, which is also reflected as systemic oxidant stress.


The 57 kb deletion in cystinosis patients extends into TRPV1 causing dysregulation of transcription in peripheral blood mononuclear cells.
Cystinosis is an autosomal recessive disease characterised by the abnormal accumulation of lysosomal cystine. Mutations in the cystinosin gene (CTNS) represent known causes for the disease. The major cystinosis mutation is a 57 kb deletion on human chromosome 17p13 that removes the majority of CTNS and the entire adjacent gene, CARKL/SHPK. In order to identify other genes that may influence the cystinosis pathobiological pathway, peripheral blood mononuclear cells (PBMC) were collected from cystinosis family members, and DNA and RNA extracted. Using whole genome transcriptional profiling, transient receptor potential vanilloid 1 (TRPV1) was found to be differentially expressed in association with cystinosis. This was verified using TaqMan qRT-PCR. There was a 72% reduction in PBMC TRPV1 mRNA levels in cystinosis individuals homozygous for the 57 kb deletion (n=6) compared to unaffected individuals without the deletion (n=6) (p=0.002). TRPV1 is a sensory receptor located on chromosome 17p13, adjacent to CARKL/SHPK. It was ascertained that the 57 kb deletion extends from exon 10 of CTNS, upstream through CARKL/SHPK, to intron 2 of TRPV1, thus deleting the first two non-coding exons. This is the first study to report that the 57 kb deletion extends into the TRPV1 gene causing dysregulation of transcription in PBMC isolated from cystinosis patients.


Signal transducer and activator of transcription 5a influences mammary epithelial cell survival and tumorigenesis.
The mammary gland undergoes extensive tissue remodeling and cell death at the end of lactation in a process known as involution. We present evidence that the prolactin-activated transcription factor signal transducer and activator of transcription 5a (Stat5a) has a crucial role in the regulation of cell death during mammary gland involution. In a transforming growth factor-alpha transgenic mouse model that exhibited delayed mammary gland involution, the absence of Stat5a facilitated involution-associated changes in morphology of the gland and the extent and timing of programmed cell death. These Stat5a-dependent changes also affected epidermal growth factor receptor-initiated mammary gland tumorigenesis. Overexpression of the transforming growth factor alpha transgene in the mammary epithelium reproducibly generated mammary hyperplasia and tumors. In the presence of the activated epidermal growth factor receptor, deletion of Stat5a delayed initial hyperplasia and mammary tumor development by 6 weeks. These observations demonstrate that Stat5a is a survival factor, and its presence is required for the epithelium of the mammary gland to resist regression and involution-mediated apoptosis. We also suggest that Stat5a is one of the antecedent, locally acting molecules that initiate the process of epithelial regression and reorganization during involution.


Sleep and wakefulness in Drosophila melanogaster.
Sleep is present and tightly regulated in every vertebrate species in which it has been carefully investigated, but what sleep is for remains a mystery. Sleep is also present in invertebrates, and an extensive analysis in Drosophila melanogaster has shown that sleep in fruit flies shows most of the fundamental features that characterize sleep in mammals. In Drosophila, sleep consists of sustained periods of quiescence associated with an increased arousal threshold. Fly sleep is modulated by several of the same stimulants and hypnotics that affect mammalian sleep. Moreover, like in mammals, fly sleep shows remarkable interindividual variability. The expression of several genes involved in energy metabolism, synaptic plasticity, and the response to cellular stress varies in Drosophila between sleep and wakefulness, and the same occurs in rodents. Brain activity also changes in flies as a function of behavioral state. Furthermore, Drosophila sleep is tightly regulated in a circadian and homeostatic manner, and the homeostatic regulation is largely independent of the circadian regulation. After sleep deprivation, recovery sleep in flies is longer in duration and more consolidated, indicated by an increase in arousal threshold and fewer brief awakenings. Finally, sleep deprivation in flies impairs vigilance and performance. Because of the extensive similarities between flies and mammals, Drosophila is now being used as a promising model system for the genetic dissection of sleep. Over the last few years, mutagenesis screens have isolated several short sleeping mutants, a demonstration that single genes can have a powerful effect on a complex trait like sleep.


Heparanase-2 expression in normal ovarian epithelium and in benign and malignant ovarian tumors.
Studies have highlighted the changes that take place in the environment between the cell and the extracellular matrix during the process of neoplastic expansion. Several papers have associated the expression of heparanase 1 with various malignant tumors. Heparanase 2 is probably related to loss of cell adhesion. The aim of this study was to evaluate the expression of heparanase 2 in epithelial neoplasia of the ovaries and in samples of normal ovarian tissue. Seventy-five ovary specimens were analyzed and divided into 3 groups: 23 malignant and 35 benign epithelial ovarian neoplasia and 17 without ovarian disease. We used 2 methodological techniques for evaluating the immunoexpression of heparanase 2. The first followed the qualitative criterion of positive or negative in relation to enzymatic expression, and the second involved computerized quantification of this expression, performed on the same slides. In the quantitative analysis, we found positivity indices for heparanase 2 expression of 72.2% and 87.3% in the samples of benign and malignant neoplasias, respectively. In these, the intensity of expression and the expression index were 147.2 and 121.2, respectively, for the benign neoplasia and 134.1 and 118.0 for the malignant neoplasia. Qualitatively, its expression was strong or moderate in 44.2% of the benign and 78.2% of the malignant tumors; its expression in all of the nonneoplastic samples was negative, with the exception of one that was weakly positive. Heparanase 2 is involved in neoplastic proliferation, but it was not exclusively associated with the malignant process. Furthermore, there was no difference in its expression between benign and malignant ovarian epithelial neoplasia.


The blood coagulation cascade.
The biochemistry of blood coagulation has been well defined over the past 50 years. Although much is known about the sequence of the proteolytic cascade and its regulation in the pathway to fibrin generation, many important questions remain unsolved about the mechanism of initiation and the structure of the protein complexes that form during blood coagulation. This article summarizes some of the advances that have been made in this field from the last quarter of 2002 and during 2003. The papers, which vary in rigor and content, have been selected on the basis of their interest and possible contribution to knowledge in this field. Summaries are given of new findings on the source of factor V and the synthesis of factor VIII, the mechanism of tissue factor action in the initiation of blood coagulation, the structure and membrane-binding properties of the protein complexes formed, and regulation of the blood coagulation cascade. Continued progress in this field offers opportunity for understanding the basis of thrombotic diseases and bleeding disorders, with the potential for defining novel targets for therapeutic applications. Some of the conclusions reviewed are conflicting, and further work will be necessary to place the results in the context of what has already been established. The structural biology of the coagulation proteins and understanding of hemostasis and thrombosis in a physiologic context have important implications for future work.


Longitudinal assessment of childhood optic gliomas: relationship between flicker visual evoked potentials and magnetic resonance imaging findings.
The aim of this study was to evaluate longitudinally functional and neuro-radiologic findings in childhood optic gliomas (OG), by comparing flicker visual evoked potentials (F-VEPs) with brain magnetic resonance imaging (MRI) changes. Fourteen children (age range: 1-13 years) with OGs underwent serial F-VEP, MRI and neuro-ophthalmic examinations over a 38 month (median, range: 6-76) follow-up. F-VEPs were elicited by 8 Hz sine-wave flicker stimuli presented in a mini-Ganzfeld. Contrast-enhanced MRI examinations were performed. Results of both tests were blindly assessed by independent evaluators. F-VEPs were judged to be improved, stable or worsened if changes in the amplitude and/or phase angle of the response exceeded the limits of test-retest variability (+/-90th percentile) established for the same patients. MRI results were judged to show regression, stabilization or progression of OG based on its changes in size (+/-20%) or extension. Two to seven pairs of F-VEP/MRI examinations per patient (median: 4) were collected. Based on a total of 38 pairs of F-VEP/MRI examinations, both tests agreed in showing worsening (progression), stabilization and improvement (regression) in 5, 15 and 10 cases, respectively. In 3 cases, F-VEPs showed a worsening and MRI a stabilization, while in 5 cases F-VEPs showed an improvement and MRI a stabilization. Agreement between F-VEP and MRI changes was 78.9% (95% CI: +/- 37%, K statistics = 0.67, P < 0.001). The results indicate that longitudinal F-VEP changes can predict changes in MRI-assessed OG size and extension, providing a non-invasive functional assay, complementary to neuro-imaging, for OG follow-up.


Identification of Climatic Factors Affecting the Epidemiology of Human West Nile Virus Infections in Northern Greece.
Climate can affect the geographic and seasonal patterns of vector-borne disease incidence such as West Nile Virus (WNV) infections. We explore the association between climatic factors and the occurrence of West Nile fever (WNF) or West Nile neuro-invasive disease (WNND) in humans in Northern Greece over the years 2010-2014. Time series over a period of 30 years (1979-2008) of climatic data of air temperature, relative humidity, soil temperature, volumetric soil water content, wind speed, and precipitation representing average climate were obtained utilising the ECMWF's (European Centre for Medium-Range Weather Forecasts) Re-Analysis (ERA-Interim) system allowing for a homogeneous set of data in time and space. We analysed data of reported human cases of WNF/WNND and Culex mosquitoes in Northern Greece. Quantitative assessment resulted in identifying associations between the above climatic variables and reported human cases of WNF/WNND. A substantial fraction of the cases was linked to the upper percentiles of the distribution of air and soil temperature for the period 1979-2008 and the lower percentiles of relative humidity and soil water content. A statistically relevant relationship between the mean weekly value climatic anomalies of wind speed (negative association), relative humidity (negative association) and air temperature (positive association) over 30 years, and reported human cases of WNF/WNND during the period 2010-2014 could be shown. A negative association between the presence of WNV infected Culex mosquitoes and wind speed could be identified. The statistically significant associations could also be confirmed for the week the WNF/WNND human cases appear and when a time lag of up to three weeks was considered. Similar statistically significant associations were identified with the weekly anomalies of the maximum and minimum values of the above climatic factors. Utilising the ERA-Interim re-analysis methodology it could be shown that besides air temperature, climatic factors such as soil temperature, relative humidity, soil water content and wind speed may affect the epidemiology of WNV.


Heart failure with recovered ejection fraction: Clinical characteristics, determinants and prognosis. CARDIOCHUS-CHOP registry.
The magnitude and the prognostic impact of recovering left ventricular ejection fraction (LVEF) in patients with heart failure (HF) and systolic dysfunction is unclear. The aim of this study was to evaluate the clinical characteristics and prognosis of patients with HFrecEF in an HF population. 449 consecutive patients were selected with the diagnosis of HF and an evaluation of LVEF in the 6 months prior to selection who were referred to two HF units. Patients with systolic dysfunction were only considered if a second echocardiogram was performed during the follow-up. At the time of diagnosis, 207 patients had LVEF > 40% (HFpEF) and 242 had LVEF ≤ 40% (HFrEF). After 1 year, the LVEF was re-evaluated in all 242 patients with a LVEF ≤ 40%: in 126 (52%), the second LVEF was > 40% (HFrecEF), and the remaining 116 (48%) had LVEF ≤ 40% (HFrEF). After 1800 ± 900 days of follow-up patients with recovered LVEF had a significantly lower mortality rate (HFpEF vs. HFrecEF: hazard ratio [HR] = 2.286, 95% confidence interval [95% CI] 1.264-4.145, p = 0.019; HFrEF vs. HFrecEF: HR = 2.222, 95% CI 1.189-4.186, p < 0.001) and hospitalization rate (HFpEF vs. HFrecEF: HR = 1.411, 95% CI 1.046-1.903, p = 0.024; HFrEF vs. HFrecEF: HR = 1.388, 95% CI 1.002-1.924, p = 0.049). The following are predictors of LVEF recovery: younger age, lower functional class, treatment with renin-angiotensin-aldosterone system inhibitors and beta-blockers, absence of defibrillator use, and non-ischemic etiology. Patients with HF and reduced LVEF who were re-evaluated after 1 year, had significant improvement in their LVEF and had a more favourable prognosis than HF with preserved and reduced ejection fraction.


Intraperitoneal and intragastric norepinephrine instillation in massive gastrointestinal bleeding--clinical study. Preliminary report.
Intraperitoneal and intragastric instillation of norepinephrine in high concentration can arrest bleeding of various origins in the gastrointestinal tract. Of the 21 cases studied, the hemorrhage was controlled in 80% of cases. This form of therapy is simple, prompt, safe and can be performed as a bed-side therapeutic procedure to stop life-threatening gastrointestinal tract bleeding. Intraperitoneal instillation of norepinephrine should not be used more than once, if the bleeding is not controlled. Intragastric norepinephrine instillations may be repeated any number of times if required.


Three-Dimensional Modeling of Solid Tumors and Their Microenvironment to Evaluate T Cell Therapy Efficacy In Vitro.
Immunotherapy development for solid tumors remains challenging, partially due to a lack of reproducible, cost-effective in vitro three-dimensional (3D) models to mimic the heterogeneous and complex tumor microenvironment. Here, we investigate the cellular anti-tumor reactivity of αβ T cells engineered to express a defined γδ TCR (TEG A3). For that purpose, we developed a 3D cytotoxicity assay targeting cell line-derived spheroids or patient-derived tumor organoids formed in serum-free media. Tumor cell lysis by TEG A3 was monitored using the Incucyte S3 live-cell imaging system with the apoptosis marker caspase 3/7 green and endpoint readouts of IFN-γ secretion in the supernatant. The 3D cytotoxicity assay model system was able to adequately demonstrate TEG A3 reactivity toward targets expressing an isoform of CD277 (CD277J). To obtain a more complex heterogeneous tumor microenvironment, patient-derived organoids were mixed with unmatched patient-derived fibroblasts or matched cancer-associated fibroblasts. In all assays, we demonstrated the tumor target specificity of TEG A3, lysing tumor cells within 48 h. Our study demonstrates the utility of complex 3D cytotoxicity assay model systems incorporating the tumor microenvironment in the functional evaluation of T cell-based adoptive immunotherapy, providing a useful platform for early-stage preclinical development of immunotherapies.


A comparison of preoperative and postoperative nutritional states of lung transplant recipients.
Malnutrition is a documented problem in some types of endstage lung disease (ESLD). Recently, isolated lung transplants have successfully reversed the respiratory failure of patients suffering from ESLD. In this study, we compare the preoperative and postoperative nutritional states of lung transplant recipients using weight-to-height ratios, anthropometric measurements, subjective global assessment, and biochemical blood values. Patients with emphysema, cystic fibrosis, and other types of bronchiectasis, but not patients with pulmonary fibrosis or pulmonary hypertension, were malnourished preoperatively. All groups had normal biochemical profiles. Caloric intake of patients with cystic fibrosis and bronchiectasis was increased above predicted basal energy expenditure levels. By six months to one year postoperatively, all groups of malnourished patients had significantly improved their nutritional status. Emphysema patients improved nutrition by maintaining preoperative caloric intake levels--however, both cystic fibrosis and bronchiectasis patients were able to achieve the same goal with significantly decreased caloric intakes. We conclude that malnourished ESLD patients receiving isolated lung grafts are able to achieve normal nutrition within one year posttransplant. Since this occurs in all cases with a reduced, or at best maintained, caloric intake, more study is needed to elucidate the factors that contribute to ESLD malnutrition.


Immune responses against human papillomavirus (HPV) type 16 virus-like particles in a cohort study of women with cervical intraepithelial neoplasia. I. Differential T-helper and IgG responses in relation to HPV infection and disease outcome.
T-helper (Th) cell-dependent IL-2 production and plasma IgG responses to virus-like particles consisting of the human papillomavirus type 16 (HPV-16) major capsid protein L1 (L1-VLP) were determined in patients with cytological evidence of cervical intraepithelial neoplasia (CIN) participating in a non-intervention prospective cohort study. IgG responses were associated with HPV-16 persistence and high-grade CIN lesions, while high frequencies of Th responses were observed in patients with both virus clearance and virus persistence, irrespective of CIN grade. The IgG response was found in conjunction with an IL-2 response to L1-VLP in 87% of the patients. Recognition of the HPV-16 L1 Th epitope (amino acids 311-335) was found to be more closely associated than recognition of L1-VLP as a whole to HPV exposure and CIN development. Among the HPV-16+ patients included in this study, those showing a Th response to amino acids 311-335 were more likely to carry the HLA DRB1*11/DQB1*0301 haplotype, while those showing an IgG response to L1-VLP were more likely to carry DRB1*0101/DQB1*0501. However, neither cell-mediated nor humoral immune responses against HPV-16 L1 appear to be sufficient for the natural control of HPV infection and CIN development.


In vitro production of glycosaminoglycans by retinal microvessel cells and lens epithelium.
Analysis of bovine lens capsules and retinal microvessel basement membranes by cellulose acetate electrophoresis, together with treatment by specific enzymes or by nitrous acid, reveals that heparan sulfate is the only demonstrable glycosaminoglycan (GAG), consistent with the observations of most other investigators on basement membrane preparations from several different tissues. When bovine retinal microvessel pericytes or endothelial cells or bovine lens epithelial cells are grown in culture in the presence of [35S]-sulfate or [3H]-glucosamine, however, both cellulose acetate electrophoresis and gel filtration chromatography reveal that the cultured pericytes synthesize primarily chondroitin sulfate, whereas the lens epithelial cells and microvascular endothelial cells produce a mixture of GAGs consisting of approximately 60% heparan sulfate and 40% chondroitin sulfate. The chondroitin sulfate chains have an Mr of 70,000, but the Mr of the heparan sulfate chains is 10,000, based on gel filtration chromatography on Sepharose CL-6B. Hence, cell culture conditions may produce phenotypic modulation of the biosynthetic capacities of these cells for GAGs. However, the difference between the in vitro and in vivo findings may in part be explained if many of the chondroitin sulfate GAGs synthesized in vitro are cell associated and are not incorporated into the basement membrane.


Clinical evaluation of the Nanoduct sweat test system in the diagnosis of cystic fibrosis after newborn screening.
After a positive newborn screening test for cystic fibrosis (CF), a sweat test is performed to confirm the diagnosis. The success rate of the generally acknowledged methods (Macroduct/Gibson and Cooke) in newborns varies between 73 and 99%. The Nanoduct sweat test system is easier to perform and less sweat is needed. The main aim of this study was to measure the success rate of the Nanoduct compared to current approved sweat test methods in a newborn population. After informed consent of the parents, newborns with a positive screening test for CF were included. The Macroduct or Gibson and Cooke and Nanoduct were performed in all infants, during the same appointment. The chloride concentration was determined by standard coulorimetry; conductivity was measured directly and converted to a NaCl molarity. One hundred eight newborns were included: 17 with CF, 7 with cystic fibrosis transmembrane regulator (CFTR)-related metabolic syndrome (CRMS), and 84 healthy children. The success rate of the Nanoduct was 93% and for the Macroduct/Gibson and Cooke 79% (McNemar, p = 0.002). The Nanoduct detected the same CF patients as the Macroduct/Gibson and Cooke; one CF patient had an equivocal result for both tests, and no patients were missed. The area under the receiver operating characteristic curve for detection of CF with the Nanoduct was 0.999, with ideal cutoff levels of 91 and 66 mmol/l, comparable to former studies. The success rate of the Nanoduct to collect sufficient sweat in infants was higher compared to the Macroduct and Gibson and Cooke.


Fragments of the earliest land plants.
The earliest fossil evidence for land plants comes from microscopic dispersed spores. These microfossils are abundant and widely distributed in sediments, and the earliest generally accepted reports are from rocks of mid-Ordovician age (Llanvirn, 475 million years ago). Although distribution, morphology and ultrastructure of the spores indicate that they are derived from terrestrial plants, possibly early relatives of the bryophytes, this interpretation remains controversial as there is little in the way of direct evidence for the parent plants. An additional complicating factor is that there is a significant hiatus between the appearance of the first dispersed spores and fossils of relatively complete land plants (megafossils): spores predate the earliest megafossils (Late Silurian, 425 million year ago) by some 50 million years. Here we report the description of spore-containing plant fragments from Ordovician rocks of Oman. These fossils provide direct evidence for the nature of the spore-producing plants. They confirm that the earliest spores developed in large numbers within sporangia, providing strong evidence that they are the fossilized remains of bona fide land plants. Furthermore, analysis of spore wall ultrastructure supports liverwort affinities.


Prader-Willi syndrome and Angelman syndrome.
Prader-Willi syndrome (PWS) and Angelman syndrome (AS) are two distinct neurogenetic disorders in which imprinted genes on the proximal long arm of chromosome 15 are affected. Although the SNORD116 gene cluster has become a prime candidate for PWS, it cannot be excluded that other paternally expressed genes in the chromosomal region 15q11q13 contribute to the full phenotype. AS is caused by a deficiency of the UBE3A gene, which in the brain is expressed from the maternal allele only. The most frequent genetic lesions in both disorders are a de novo deletion of the chromosomal region 15q11q13, uniparental disomy 15, an imprinting defect or, in the case of AS, a mutation of the UBE3A gene. Microdeletions in a small number of patients with PWS and AS have led to the identification of the chromosome 15 imprinting center (IC). The IC consists of two critical elements, which act in cis to regulate imprinting in the whole chromosome 15q11q13 imprinted domain.


Working Alliance and Stages of Change for Employment: The Intermediary Role of Autonomous Motivation, Outcome Expectancy and Vocational Rehabilitation Engagement.
Purpose Working alliance is one of the most important common factors for successful counseling/psychotherapy outcomes. Based on the empirical literature about working alliance, it seems that self-determination and self-efficacy theory (SDT/SET) can potentially be used as a motivational model to explain the relationship between working alliance and vocational rehabilitation (VR) outcomes. The purpose of this study is to evaluate three primary SDT/SET constructs, autonomous motivation, expectancy and engagement, as mediators for the relationship between working alliance and stages of change (SOC) for employment. Methods A serial multiple mediation analysis (SMMA) was computed to evaluate autonomy, outcome expectancy, and VR engagement as mediators of the relationship between working alliance and SOC for employment in a sample of 277 people with chronic illness and disability (CID) receiving services from state VR agencies in the United States. Results The SMMA results indicated that working alliance was positively associated with SOC for employment (total effect), while the direct effect between working alliance and SOC for employment was not significant after controlling for the effects of the mediators, indicating significant mediation effects. The mediation effects were estimates of the indirect effects for working alliance on SOC for employment through (a) autonomous motivation, (b) outcome expectancy, (c) VR engagement, and (d) autonomous motivation, outcome expectancy and VR engagement together. Conclusions The results indicated that a strong working alliance has the benefit of helping consumers develop autonomous motivation to work and increasing their vocational outcome expectancy and engagement in VR services, leading to employment.


[Iron metabolism and metalloproteins (review)].
Problems of ferrokinetics, participation of metalloproteins transferrin, ferritin and lactoferrin in metabolism of iron at the step of the metal absorption, transport of iron by means of transferrin, haptoglobin and hemopexin, interaction of transferrin with reticulocytes, deposition of iron in ferritin, mobilization of iron from ferritin via ceruloplasmin are considered. Importance of blood serum ferritin is discussed.