Elevated Framingham risk score in HIV-positive patients on highly active antiretroviral therapy: results from a Norwegian study of 721 subjects.
Highly active antiretroviral therapy (HAART) may induce dyslipidemia and thus increase the risk of future cardiovascular heart disease (CHD). In this cross-sectional study performed in 2000-2001, the prevalence of a Framingham CHD risk score of >20% in HIV-positive individuals treated or not treated with HAART was compared with that in age- and gender-matched controls. The study included 721 subjects: 219 HIV-positive individuals on HAART, 64 HIV-positive, HAART-naïve individuals, and 438 age- and gender-matched controls randomly selected from a simultaneous health survey. The prevalence of a 10-year estimated CHD risk of >20% was 11.9% in patients on HAART compared to 5.3% in controls ( P=0.004). The main contributors to the increased CHD risk in patients on HAART were increased prevalence of daily smoking (54.5% vs 30.1%; P<0.001), total cholesterol of >6.2 mmol/l (36.1% vs 21.7%; P<0.001), and HDL cholesterol of < 0.9 mmol/l (20.9% vs 8.0%; P<0.001). In HAART-naïve patients, the prevalence of a 10-year estimated CHD risk of >20% was 6.3% ( P=0.25 vs HAART patients, P=0.76 vs controls), the prevalence of daily smoking was 56.3% ( P=0.89 vs HAART patients, P<0.001 vs controls), the prevalence of total cholesterol >6.2 mmol/l was 9.4% ( P<0.001 vs HAART patients, P=0.019 vs controls), and the prevalence of HDL cholesterol of <0.9 mmol/l was 30.9% ( P=0.16 vs HAART patients, P<0.001 vs controls). The results show that, compared to controls, twice as many patients on HAART have an estimated 10-year CHD risk above 20%. These patients are candidates for intensive interventions. HAART patients should be encouraged to permanently stop smoking, make healthy food choices, and increase physical activity. In patients with elevated lipid levels, a change in the HAART regimen or treatment with lipid-lowering drugs should be considered.


Advance directives and personal identity: what is the problem?
The personal identity problem expresses the worry that due to disrupted psychological continuity, one person's advance directive could be used to determine the care of a different person. Even ethicists, who strongly question the possibility of the scenario depicted by the proponents of the personal identity problem, often consider it to be a very potent objection to the use of advance directives. Aiming to question this assumption, I, in this paper, discuss the personal identity problem's relevance to the moral force of advance directives. By putting the personal identity argument in relation to two different normative frameworks, I aim to show that whether or not the personal identity problem is relevant to the moral force of advance directives, and further, in what way it is relevant, depends entirely on what normative reasons we have for respecting advance directives in the first place.


Fertility recovery and pregnancy after allogeneic hematopoietic stem cell transplantation in Fanconi anemia patients.
Reduced fertility is one clinical manifestation among other well known Fanconi anemia features. Most recipients of allogeneic hematopoietic stem cell transplantation suffer from secondary infertility owing to gonadal damage from myeloablative conditioning. In order to evaluate the rate of pregnancy in Fanconi anemia transplanted patients, we performed a retrospective analysis of female patients transplanted in 15 centers from 1976 to 2008. Among 578 transplanted Fanconi anemia patients, we identified 285 transplanted females of whom 101 patients were aged 16 years or over. Ten became pregnant (4 twice). Before hematopoietic stem cell transplantation all had confirmed Fanconi anemia diagnosis. Median age at transplantation was 12 years (range 5-17 years). Conditioning regimen consisted of cyclophosphamide with or without irradiation. During follow up, 5 of 10 patients presented signs of ovarian failure. Among those, 2 patients spontaneously recovered regular menses, and 3 received hormonal replacement therapy. Pregnancy occurred from four to 17 years after hematopoietic stem cell transplantation. Three patients had preterm deliveries, one patient had a hysterectomy for bleeding. All 14 newborns had normal growth and development without congenital diseases. In conclusion, recovery of normal ovarian function and a viable pregnancy is a realistic but relatively rare possibility even in Fanconi anemia patients following hematopoietic stem cell transplantation. Mechanisms of fertility recovery are discussed.


Assessing multiscale complexity of short heart rate variability series through a model-based linear approach.
We propose a multiscale complexity (MSC) method assessing irregularity in assigned frequency bands and being appropriate for analyzing the short time series. It is grounded on the identification of the coefficients of an autoregressive model, on the computation of the mean position of the poles generating the components of the power spectral density in an assigned frequency band, and on the assessment of its distance from the unit circle in the complex plane. The MSC method was tested on simulations and applied to the short heart period (HP) variability series recorded during graded head-up tilt in 17 subjects (age from 21 to 54 years, median = 28 years, 7 females) and during paced breathing protocols in 19 subjects (age from 27 to 35 years, median = 31 years, 11 females) to assess the contribution of time scales typical of the cardiac autonomic control, namely in low frequency (LF, from 0.04 to 0.15 Hz) and high frequency (HF, from 0.15 to 0.5 Hz) bands to the complexity of the cardiac regulation. The proposed MSC technique was compared to a traditional model-free multiscale method grounded on information theory, i.e., multiscale entropy (MSE). The approach suggests that the reduction of HP variability complexity observed during graded head-up tilt is due to a regularization of the HP fluctuations in LF band via a possible intervention of sympathetic control and the decrement of HP variability complexity observed during slow breathing is the result of the regularization of the HP variations in both LF and HF bands, thus implying the action of physiological mechanisms working at time scales even different from that of respiration. MSE did not distinguish experimental conditions at time scales larger than 1. Over a short time series MSC allows a more insightful association between cardiac control complexity and physiological mechanisms modulating cardiac rhythm compared to a more traditional tool such as MSE.


From powerhouse to processing plant: conserved roles of mitochondrial outer membrane proteins in tRNA splicing.
The mitochondrial cytoplasmic surface serves as a processing site for numerous RNAs from budding yeast to metazoans. We report that budding yeast mitochondrial outer membrane (MOM) proteins that are subunits of the translocase of the outer mitochondrial membrane (Tom70 and Tom 22) and sorting and assembly machinery (Sam37) are required for efficient pretransfer RNA (pre-tRNA) splicing. Defective pre-tRNA splicing in MOM mutants is due not to loss of respiratory metabolism but instead inefficient targeting/tethering of tRNA splicing endonuclease (SEN) subunits to mitochondria. Schizosaccharomyces pombe SEN subunits also localize to mitochondria, and Tom70 is required for this localization and pre-tRNA splicing. Thus, the role of MOM protein in targeting/tethering SEN subunits to mitochondria has been conserved for >500 million years.


Algae harvesting for biofuel production: influences of UV irradiation and polyethylenimine (PEI) coating on bacterial biocoagulation.
There is a pressing need to develop efficient and sustainable separation technologies to harvest algae for biofuel production. In this work, two bacterial species (Escherichia coli and Rhodococus sp.) were used as biocoagulants to harvest Chlorella zofingiensis and Scenedesmus dimorphus. The influences of UV irradiation and polyethylenimine (PEI)-coating on the algal harvesting efficiency were investigated. Results showed that the UV irradiation could slightly enhance bacteria-algae biocoagulation and algal harvesting efficiency. In contrast, the PEI-coated E. coli cells noticeably increased the harvesting efficiencies from 23% to 83% for S. dimorphus when compared to uncoated E. coli cells. Based on the soft-particle Derjaguin-Landau-Verwey-Overbeek (DLVO) theory, an energy barrier existed between uncoated E. coli cells and algal cells, whereas the PEI coating on E. coli cells eliminated the energy barrier, thereby the biocoagulation was significantly improved. Overall, this work presented groundwork toward the potential use of bacterial biomass for algal harvesting from water.


Impact of lung cancer screening results on smoking cessation.
Lung cancer screening programs may provide opportunities to reduce smoking rates among participants. This study evaluates the impact of lung cancer screening results on smoking cessation. Data from Lung Screening Study participants in the National Lung Screening Trial (NLST; 2002-2009) were used to prepare multivariable longitudinal regression models predicting annual smoking cessation in those who were current smokers at study entry (n = 15489, excluding those developing lung cancer in follow-up). The associations of lung cancer screening results on smoking cessation over the trial period were analyzed. All hypothesis testing used two sided P values. In adjusted analyses, smoking cessation was strongly associated with the amount of abnormality observed in the previous year's screening (P < .0001). Compared with those with a normal screen, individuals were less likely to be smokers if their previous year's screen had a major abnormality that was not suspicious for lung cancer (odds ratio [OR] = 0.811; 95% confidence interval [CI] = 0.722 to 0.912; P < .001), was suspicious for lung cancer but stable from previous screens (OR = 0.785; 95% CI = 0.706 to 0.872; P < .001), or was suspicious for lung cancer and was new or changed from the previous screen (OR = 0.663; 95% CI = 0.607 to 0.724; P < .001). Differences in smoking prevalence were present up to 5 years after the last screen. Smoking cessation is statistically significantly associated with screen-detected abnormality. Integration of effective smoking cessation programs within screening programs should lead to further reduction in smoking-related morbidity and mortality.


Exosomes-derived miR-548am-5p promotes colorectal cancer progression.
Exosomes are vital modulators in intercellular communication and microRNAs (miRNAs) are enriched within exosomes. MiRNAs are important participants in affecting colorectal cancer (CRC) progression, but the influence and latent mechanism of cancer-secreted exosomal miRNAs in colorectal cancer are not fully understood. miR-548am-5p has been reported to be differentially expressed in colon cancer and is indicated as a biomarker for colon cancer diagnosis at the early stage. In this study, we aimed to explore the role of exosomes-derived miR-548am-5p in CRC development. ISH and FISH were implemented to assess miR-548am-5p expression and location in CRC. CRC cells-secreted exosomes were identified via transmission electron microscopy and western blot. Colony formation, sphere formation and flow cytometry assessed the changes in proliferation, stemness and apoptosis of CRC cells. Bioinformatic analyses and mechanical experiments verified the binding of miR-548am-5p and RAR-related orphan receptor A (RORA). Our study identified miR-548am-5p was highly expressed in CRC tissues and cells. Tumor-derived exosomes expedited CRC cell proliferation and stemness along with secreted miR-548am-5p. Moreover, miR-548am-5p inhibition suppressed CRC cell proliferation and stemness while promoting cell apoptosis. RORA was the target mRNA of miR-548am-5p. Down-regulation of RORA was discovered in CRC and its expression was repressed by CRC cell-derived exosomes. As a result, our work elucidated that tumor-derived exosomal miR-548am-5p promoted CRC cell proliferation and stemness via targeting RORA, providing a valuable sight for CRC therapy.


Quantitative assessment of image quality enhancement due to unsharp-mask processing in x-ray fluoroscopy.
Spatial unsharp-mask processing and its variants are commonly used in x-ray radiography to enhance image contrast. We investigated the effect of three unsharp-masking filter kernels of different sizes on the detection of an advanced guidewire tip in simulated x-ray fluoroscopy image sequences. To isolate the effect of visual temporal processing, we repeated the experiments on single images. Filter gains were selected so that all three kernels increased the contrast of a 0.018-in. (0.457-mm) guidewire by a factor of 2 but had different effects on image noise and signal profiles. There was no statistically significant effect of unsharp masking on human-observer performance in single images. However, all three kernels significantly improved average performance in image sequences, and the guidewire contrast required for detection was reduced by 32%-40%. A prewhitening channelized observer model predicted the disparity between sequences and single images and fitted measurements at different kernel sizes well. A nonprewhitening observer model did not. We conclude that unsharp masking is a simple and effective method of improving guidewire visualization in fluoroscopically guided interventional procedures and that quantitative image quality studies are essential for evaluation of image-processing techniques in sequences such as x-ray fluoroscopy.


The hazards of honey: infantile botulism.
Infantile botulism is a rare cause of neuromuscular weakness resulting from ingestion of Clostridium botulinum-an anaerobic Gram-positive bacillus found universally in soil. The only definite food source known to cause infantile botulism is honey; previously, links to formula milk have been postulated but not definitely sourced. We present an interesting case report of a 2-month-old infant with this rare condition, including the diagnostic difficulties that ensued. A brief overview of the condition follows. This is the first case in the UK in which C botulinum was successfully isolated from both the patient and the suspected source-a jar of honey. The importance of food labelling as a public health message is highlighted.


Speciation in the White-breasted Nuthatch (Sitta carolinensis): a multilocus perspective.
Inferring the evolutionary and ecological processes that have shaped contemporary species distributions using the geographic distribution of gene lineages is the principal goal of phylogeographic research. Researchers in the field have recognized that inferences made from a single gene, often mitochondrial, can be informative regarding the pattern of diversification but lack conclusive information regarding the evolutionary mechanisms that led to the observed patterns. Here, we use a multilocus (20 loci) data set to explore the evolutionary history of the White-breasted Nuthatch (Sitta carolinensis). A previous single-locus study found S. carolinensis is comprised of four reciprocally monophyletic clades geographically restricted to the pine and oak forests of: (i) eastern North America, (ii) southern Rocky Mountain and Mexican Mountain ranges, (iii) Eastern Sierra Nevada and Northern Rocky Mountains and (iv) Pacific slope of North America. The diversification of the clades was attributed to the fragmentation of North American pine and oak woodlands in the Pliocene with subsequent divergences owing to the Pleistocene glacial cycles. Principal component, clustering and species tree analyses of the multilocus data resolved the same four groups or lineages found in the single-locus study. Coalescent analyses and hypothesis testing of nested isolation and migration models indicate that isolation and not gene flow has been the major evolutionary mechanism responsible for shaping genetic variation, and all the divergence events within S. carolinensis have occurred in response to the Pleistocene glacial cycles.


Projections from subnucleus oralis of the spinal trigeminal nucleus to contralateral thalamus via the relay of juxtatrigeminal nucleus and dorsomedial part of the principal sensory trigeminal nucleus in the rat.
Following injection of HRP into contralateral thalamus, retrogradely labeled cells were observed in principal sensory trigeminal nucleus (Vp) and an area of juxtatrigeminal nucleus (JX) formerly described by John and Tracey (1987). When PHA-L was delivered to dorsomedial part of the subnucleus oralis (Vodm), PHA-L labeled terminals were seen in dorsomedial part of the Vp (Vpdm) and in the JX region. Comparing the distribution of PHA-L labeled terminal field with that of HRP labeled JX neurons showed that the labeled terminals and neurons were overlapped closely in the JX. The distribution patterns of the labeled terminals and JX neurons were also the same: viewed on the coronal planes caudal-rostrally, both of the labelings began to appear at the levels where the facial nerve root was just broken. Rostrally, at middle levels of the motor trigeminal nucleus (Vmo), the labelings showed their typical view covering dorsal and ventral JX (dJX, vJX). The labelings disappeared at rostral poles of the Vmo and Vp. When injections of PHA-L into the Vodm and HRP into the contralateral thalamus was made in one rat, the contacts between Vodm projecting terminals labeled with PHA-L and HRP labeled trigemino-thalamic neurons were seen in the JX and also in the Vpdm. Then, electron microscopic (EM) study was done, injections of kainic acid into the Vodm and HRP into the contralateral thalamus was performed simultaneously. After EM embedding, the JX and Vpdm regions were selected, ultrathin sections were cut and observed with EM. In both areas, axo-somatic and axo-dendritic synapses were seen between degenerated boutons and HRP labeled somata or dendrites. Namely, the Vodm projecting terminals synapsed on trigemino-thalamic neurons in the JX and Vpdm. Anyway, axo-dendritic synapses was the main type of observed synapses. Thus, the present work demonstrated 1. the JX containing a group of trigemno-thalamic neurons was a target of special projections froin the Vodm; 2. The Vodm neurons projected to the contralateral thalamus through the relay of JX and Vpdm neurons.


Connected speech intelligibility of children with cochlear implants and children with normal hearing.
The objective of this study was to compare the connected speech intelligibility of children who use cochlear implants with that of children who have normal hearing. Previous research has shown that speech intelligibility improves from before cochlear implantation to after implantation and that the speech intelligibility of children who use cochlear implants compares favorably with that of children who use conventional hearing aids. However, no research has yet addressed the question of how the speech intelligibility of children who use cochlear implants compares to that of children with normal hearing. In the current study, archival data on connected speech intelligibility from 51 children with cochlear implants were compared with newly collected data from 47 children with normal hearing. Results showed that for children with cochlear implants, greater intelligibility was associated with both increased chronological age and increased duration of cochlear implant use. Consistent with previous studies, children with normal hearing achieved adult-like or near-adult-like intelligibility around the age of 4 years, but a similar peak in intelligibility was not observed for the children who used cochlear implants. On the whole, children with cochlear implants were significantly less intelligible than children with normal hearing, when controlling both for chronological age and for length of auditory experience. These results have implications for the socialization and education of children with cochlear implants, particularly with respect to on-time placement in mainstream educational environments with age peers.


Mammalian PIG-L and its yeast homologue Gpi12p are N-acetylglucosaminylphosphatidylinositol de-N-acetylases essential in glycosylphosphatidylinositol biosynthesis.
Glycosylphosphatidylinositol (GPI) is used as a membrane anchor by many eukaryotic cell-surface proteins. The second step of GPI biosynthesis is de-N-acetylation of N-acetylglucosaminylphosphatidylinositol (GlcNAc-PI). We have previously cloned the rat PIG-L gene by expression cloning that complemented a mutant Chinese hamster ovary cell line defective in this step. Here we show that recombinant rat PIG-L protein purified from Escherichia coli as a complex with GroEL has GlcNAc-PI de-N-acetylase activity in vitro. The activity was not enhanced by GTP, which is known to enhance the de-N-acetylase activity of mammalian cell microsomes. As with other de-N-acetylases that act on the GlcNAc moiety, metal ions, in particular Mn2+ and Ni2+, enhanced the enzyme activity of PIG-L. The Saccharomyces cerevisiae YMR281W open reading frame encodes a protein (termed Gpi12p) with 24% amino acid identity with rat PIG-L. On transfection into mammalian PIG-L-deficient cells, this gene, GPI12, restored the cell-surface expression of GPI-anchored proteins and GlcNAc-PI de-N-acetylase activity. The disruption of the gene caused lethality in S. cerevisiae. These results indicate that GlcNAc-PI de-N-acetylase is conserved between mammals and yeasts and that the de-N-acetylation step is also indispensable in yeasts.


[Expression of Osterix mRNA and protein induced by recombinant human bone morphogenetic protein-2 in human periodontal ligament cells].
To detect the changes of Osterix (Osx) mRNA and protein expression in human periodontal ligament cells (HPDLCs) induced by recombinant human bone morphogenetic protein-2 (rhBMP-2), and examine the role of BMP-2 and Osx during the osteogenic differentiation of HPDLCs. HPDLCs were isolated and cultured in vitro with explant method. Cells at passage 3 were cultured in DMEM with rhBMP-2 at different concentrations (50, 100, 150, 200, 250, 300, 400, and 600 µg/L) for different times (2, 3, 5, 7, 10, 14 and 21 days). Then the expressions of Osx mRNA and protein were measured by real-time reverse transcription (RT)-PCR and Western blotting respectively. Cells were treated with 10 µmol/L SB203580 (p38 inhibitor) to inhibit p38 phosphorylation induced by rhBMP-2. The mineralization nodules formation and the expressions of phosphorylated p38 and Osx mRNA were detected respectively. During the culture of rhBMP-2, the expression of Osx mRNA significantly increased. Initially Osx protein had a low expression and then increased in a time-dependent manner followed by the production of bone-like nodules in HPDLCs. Under the effect of SB203580, the up-regulation of phosphorylated p38 expression induced by rhBMP-2 was significantly inhibited as well as the expression of Osx (Osx mRNA expression: 0.378 ± 0.034 vs 0.134 ± 0.027, Osx protein expression: 0.353 ± 0.024 vs 0.155 ± 0.031, both P < 0.01). Meanwhile the mineralization nodules formed by HPDLCs induced by rhBMP-2 were fewer and delayed. BMP-2 has a significant positive regulatory role on the expression of Osx in HPDLCs. And p38 pathway is an important link of this regulatory process. Thus, as an important signaling pathway in osteogenic differentiation of HPDLCs, BMP-2/p38/Osx may be involved in periodontal tissue remodeling.


Drug targets in stress-related disorders.
Nervous and immune systems mutually cooperate via release of mediators of both neurological and immunological derivation. Adrenocorticotropin hormone (ACTH) is a product of the hypothalamus-pituitary adrenal axis (HPAA) which stimulates secretion of corticosteroids from adrenals. In turn, corticosteroids modulate the immune response in virtue of their anti-inflammatory activity. On the other hand, catecholamines, products of the sympathetic nervous system (SNS), regulate immune function by acting on specific beta-adrenergic receptors. Conversely, cytokines released by monocytes/macrophages and lymphocytes, upon antigenic stimulation, are able to cross the blood-brain-barrier, thus modulating nervous functions (e.g., thermoregulation, sleep, and appetite). However, cytokines are locally produced in the brain, especially in the hypothalamus, thus contributing to the development of anorexic, pyrogenic, somnogenic and behavioural effects. Besides pathogens and/or their products, the so-called stressors are able to activate both HPAA and SNS, thus influencing immune responses. In this respect, many studies conducted in medical students taking exams have evidenced an array of stress-induced immune alterations. Phobic disorders and migraine without aura (MWA) represent examples of stress-related disorders in which phagocytic immune deficits, endotoxemia and exaggerated levels of proinflammatory cytokines [Tumor Necrosis Factor-alpha (TNF- alpha), and interleukin- 1 beta] have been detected. Quite interestingly, administration of a thymic hormone could ameliorate clinical symptoms in phobic patients. In MWA patients, a beta-blocker, propranolol, could mitigate migraine, whose cessation coincided with a drop of TNF-alpha serum concentration. In phobic disorders and in MWA, benzodiazepines are very often administered and, in this respect, some of them, such as diazepam, inhibit immune functions, while others, e.g., alprazolam, enhance immune responses. Alprazolam could improve clinical symptoms in MWA patients. Chronic Fatigue Syndrome (CFS) is a disorder whose etiology and pathogenesis are still unknown. In this syndrome both abnormalities of nervous and immune systems have been reported. Despite many immune parameters evaluated in CFS no specific biomarkers of disease have been found. Our own data are in agreement with current literature in that we found decreased levels of serum (IFN)-gamma in these patients, thus indicating a predominance of T helper (h)1 response in CFS. Also leptin, a hormone which regulates food intake, fluctuates within normal ranges in CFS individuals. Quite interestingly, in depressed patients, used as controls, leptinaemia was more elevated than in CFS. Finally, in a series of recent therapeutic trials several immunomodulating agents have been used, such as staphypan Berna, lactic acid bacteria, kuibitang and intravenous immunoglobulin. In conclusion, it seems that major drug targets in stress-related disorders are immune cells in terms of inhibition of proinflammatory cytokines and modulation of Th responses. In particular, according to recent evidences, antidepressants seem to exert beneficial effects in experimental autoimmune neuritis in rats by decreasing IFN- beta release or augmenting NK activity in depressed patients.


Early treatment of degenerative arthritis of the thumb carpometacarpal joint.
Degenerative arthritis of the thumb carpometacarpal (CMC) joint is a common disorder that may affect anyone but most frequently affects the postmenopausal female population. Because of its high prevalence, the management of the condition has been a popular topic among hand surgeons and therapists worldwide. There are many decisions to consider when devising the appropriate treatment plan for each patient. In particular, early stages of thumb CMC joint arthritis may be treated nonoperatively or with less invasive surgical techniques to relieve symptoms, restore function and strength, stop the progression of the disease, and even potentially reverse the process. This article explores treatment options at the disposal of primary care physicians and hand surgeons for early thumb CMC arthritis.


SDF-1α degrades whereas glycoprotein 120 upregulates Bcl-2 interacting mediator of death extralong isoform: implications for the development of T cell memory.
After a primary immune response, T cell memory occurs when a subset of Ag-specific T cells resists peripheral selection by acquiring resistance to TCR-induced death. Recent data have implicated Bcl-2 interacting mediator of death (Bim) as an essential mediator of the contraction phase of T cell immunity. In this article, we describe that stromal-derived factor-1α (SDF-1α) ligation of CXCR4 on activated T cells promotes two parallel processes that favor survival, phospho-inactivation of Foxo3A, as well as Bim extralong isoform (Bim(EL)) degradation, both in an Akt- and Erk-dependent manner. Activated primary CD4 T cells treated with SDF-1α therefore become resistant to the proapoptotic effects of TCR ligation or IL-2 deprivation and accumulate cells of a memory phenotype. Unlike SDF-1α, gp120 ligation of CXCR4 has the opposite effect because it causes p38-dependent Bim(EL) upregulation. However, when activated CD4 T cells are treated with both gp120 and SDF-1α, the SDF-1α-driven effects of Bim(EL) degradation and acquired resistance to TCR-induced death predominate. These results provide a novel causal link between SDF-1α-induced chemotaxis, degradation of Bim(EL), and the development of CD4 T cell memory.


The dynamic changes of DNA methylation and histone modifications of salt responsive transcription factor genes in soybean.
Epigenetic modification contributes to the regulation of gene expression and plant development under salinity stress. Here we describe the identification of 49 soybean transcription factors by microarray analysis as being inducible by salinity stress. A semi-quantitative RT-PCR-based expression assay confirmed the salinity stress inducibility of 45 of these 49 transcription factors, and showed that ten of them were up-regulated when seedlings were exposed to the demethylation agent 5-aza-2-deoxycytidine. Salinity stress was shown to affect the methylation status of four of these ten transcription factors (one MYB, one b-ZIP and two AP2/DREB family members) using a combination of bisulfite sequencing and DNA methylation-sensitive DNA gel blot analysis. ChIP analysis indicated that the activation of three of the four DNA methylated transcription factors was correlated with an increased level of histone H3K4 trimethylation and H3K9 acetylation, and/or a reduced level of H3K9 demethylation in various parts of the promoter or coding regions. Our results suggest a critical role for some transcription factors' activation/repression by DNA methylation and/or histone modifications in soybean tolerance to salinity stress.


Intraspecific phenotypic variation among alewife populations drives parallel phenotypic shifts in bluegill.
Evolutionary diversification within consumer species may generate selection on local ecological communities, affecting prey community structure. However, the extent to which this niche construction can propagate across food webs and shape trait variation in competing species is unknown. Here, we tested whether niche construction by different life-history variants of the planktivorous fish alewife (Alosa pseudoharengus) can drive phenotypic divergence and resource use in the competing species bluegill (Lepomis macrochirus). Using a combination of common garden experiments and a comparative field study, we found that bluegill from landlocked alewife lakes grew relatively better when fed small than large zooplankton, had gill rakers better adapted for feeding on small-bodied prey and selected smaller zooplankton compared with bluegill from lakes with anadromous or no alewife. Observed shifts in bluegill foraging traits in lakes with landlocked alewife parallel those in alewife, suggesting interspecific competition leading to parallel phenotypic changes rather than to divergence (which is commonly predicted). Our findings suggest that species may be locally adapted to prey communities structured by different life-history variants of a competing dominant species.


Visual space is represented by nonmatching topographies of distinct mouse retinal ganglion cell types.
The distributions of neurons in sensory circuits display ordered spatial patterns arranged to enhance or encode specific regions or features of the external environment. Indeed, visual space is not sampled uniformly across the vertebrate retina. Retinal ganglion cell (RGC) density increases and dendritic arbor size decreases toward retinal locations with higher sampling frequency, such as the fovea in primates and area centralis in carnivores [1]. In these locations, higher acuity at the level of individual cells is obtained because the receptive field center of a RGC corresponds approximately to the spatial extent of its dendritic arbor [2, 3]. For most species, structurally and functionally distinct RGC types appear to have similar topographies, collectively scaling their cell densities and arbor sizes toward the same retinal location [4]. Thus, visual space is represented across the retina in parallel by multiple distinct circuits [5]. In contrast, we find a population of mouse RGCs, known as alpha or alpha-like [6], that displays a nasal-to-temporal gradient in cell density, size, and receptive fields, which facilitates enhanced visual sampling in frontal visual fields. The distribution of alpha-like RGCs contrasts with other known mouse RGC types and suggests that, unlike most mammals, RGC topographies in mice are arranged to sample space differentially.


[Value of Hartmann's operation as an emergency intervention in sigmoid diverticulitis].
Improvements in antibiotic and intensive care treatment have facilitated the conservative therapy of patients suffering from complicated diverticulitis of the sigmoid for the time necessary for preparation in view of early elective resection of the sigmoid colon. Consequently, early elective resection of the sigmoid colon with primary anastomosis has become very popular as the treatment of choice in complicated diverticulitis. This has led us to a retrospective evaluation of our own results with both Hartmann's operation and resection with primary anastomosis. 55 patients with complicated diverticulitis of the sigmoid colon were operated on at our institution between 1981 and 1990. Group I consisted of 33 patients (17 females and 16 males) aged 39 to 89 years who underwent Hartmann's operation. Group II included 11 patients (6 females and 5 males) aged 39 to 85 years treated by resection and primary anastomosis. The remainder of the patients was treated by a three-step procedure and should not be considered here. In group I 25 patients (76%) suffered complications either at the Hartmann's operation or at descendorectostomy or both. Mortality was 6%. Only 76% of patients had their stoma closed after an average of 3.8 months. The average hospital stay of both operations combined amounted to 61 days. In group II 3 patients had complications, one of which was lethal (lung emboly). The total hospital stay in this group averaged 22 days. For Hinchey stages I and II conservative treatment, bowel preparation and early elective resection with primary anastomosis should be attempted. In cases of general peritonitis (Stages III and IV) Hartmann's operation is still the treatment of our choice.


Estrogen mobilizes a subset of estrogen receptor-alpha-immunoreactive vesicles in inhibitory presynaptic boutons in hippocampal CA1.
Although the classical mechanism of estrogen action involves activation of nuclear transcription factor receptors, estrogen also has acute effects on neuronal signaling that occur too rapidly to involve gene expression. These rapid effects are likely to be mediated by extranuclear estrogen receptors associated with the plasma membrane and/or cytoplasmic organelles. Here we used a combination of serial-section electron microscopic immunocytochemistry, immunofluorescence, and Western blotting to show that estrogen receptor-alpha is associated with clusters of vesicles in perisomatic inhibitory boutons in hippocampal CA1 and that estrogen treatment mobilizes these vesicle clusters toward synapses. Estrogen receptor-alpha is present in approximately one-third of perisomatic inhibitory boutons, and specifically in those that express cholecystokinin, not parvalbumin. We also found a high degree of extranuclear estrogen receptor-alpha colocalization with neuropeptide Y. Our results suggest a novel mode of estrogen action in which a subset of vesicles within a specific population of inhibitory boutons responds directly to estrogen by moving toward synapses. The mobilization of these vesicles may influence acute effects of estrogen mediated by estrogen receptor-alpha signaling at inhibitory synapses.


Nano-sized molecularly imprinted polymer for selective ultrasound-assisted microextraction of pesticide Carbaryl from water samples: Spectrophotometric determination.
Ultrasound-assisted dispersive solid phase microextraction followed by UV-Vis spectrophotometry (UA-DSPME UV-Vis) was designed for the extraction and preconcentration of Carbaryl using nano-sized molecularly-imprinted polymer (MIP-NP). Nano-sized Carbaryl-imprinted polymer was characterized by scanning electron microscopy. A preliminary Plackett-Burman design was applied for screening. Subsequently, central composite design under response surface methodology was used to investigate and model the Carbaryl adsorption as response as well as to optimize this response versus variables such as Carbaryl MIP-NP mass, sonication time, temperature, eluent volume, pH and vortex time. At optimum experimental conditions, UAMSPE-UV-Vis exhibited a linear range of 0.1-1.2mgL-1. The enhancement and preconcentration factors were obtained to be 30.6 and 25.0, respectively, for the extraction of Carbaryl by MIP-NP. In addition, the values of 0.033 and 0.11mgL-1 were obtained for limit of detection and limit of quantification, respectively. The value of 4.3% determined for relative standard deviation for the separation and preconcentration of Carbaryl after 5 repetitions shows the acceptable repeatability of the process. Finally, the developed method was successfully applied for the determination of Carbaryl in water samples.


Complement-mediated acute effects of liposome-encapsulated hemoglobin.
Recent studies on liposome-encapsulated hemoglobin (LEH) have indicated that this potential blood substitute can activate the complement (C) system of rats, pigs and man. The reaction can involve both the classical and the alternative pathways, and is mediated, in part, by the binding of natural anti-lipid antibodies to the lipid membrane of liposomes. The significance of these discoveries lies in the fact that C activation appears to be the primary cause of the acute physiological, hematological and laboratory changes that have been observed previously in rats and pigs following the administration of LEH or liposomes, which changes include pulmonary vasoconstriction with decreased cardiac output. In light of the proposed use of LEH as an emergency blood substitute, the latter impairment of cardiopulmonary function may warrant particular circumspection as it could aggravate the clinical state of trauma patients who are prone to develop respiratory distress partly as a consequence of C activation by the injury. Our studies on rats and pigs suggest that the above acute side effects of LEH, including the cardiopulmonary distress, can be efficiently inhibited with soluble complement receptor type I, a specific inhibitor of C activation.


Mechanical destabilization induced by controlled annular incision of the intervertebral disc dysregulates metalloproteinase expression and induces disc degeneration.
An investigation of mechanical destabilization of the lumbar ovine intervertebral disc (IVD) inducing IVD degeneration (IVDD) as determined by multiparameter outcome measures (magnetic resonance imaging [MRI], IVD composition, biomechanical testing, gene profiling, immunohistochemistry, and immunoblotting). To assess the effect of IVD mechanical destabilization on matrix protein and metalloproteinase gene expression to investigate the pathophysiological mechanisms of lumbar IVDD. Several earlier studies have used annular transection to induce IVDD in sheep, but none have optimized or validated the most appropriate lesion size. The annulus fibrosus (AF) incision inducing maximal change in IVD biomechanics was applied to L1-L2, L3-L4, and L5-L6 discs in vivo to compare with a sham procedure at 3 months post operation. IVDs were evaluated by MRI, biomechanics, histopathology, proteoglycan and collagen content, gene expression, and aggrecan proteolysis by Western blotting. Significant changes were observed in lesion (6 × 20 mm(2)) compared with sham IVDs at 3 months post operation: reduced disc height on MRI; increased neutral zone in biomechanical testing; depleted proteoglycan and collagen content in the nucleus pulposus (NP) and lesion half of the AF but not in the contralateral AF; increased messenger RNA for collagen I and II, aggrecan, versican, perlecan, matrix metalloproteinase (MMP)-1 & 13, and ADAMTS-5, in the lesion-site AF and NP but not in the contralateral AF. ADAMTS-4 messenger RNA was increased in the lesion-site AF but decreased in the NP. Despite an upregulation in MMPs, there was no change in MMP- or ADAMTS-generated aggrecan neoepitopes in any region of the IVD in lesion or sham discs. Lumbar IVDD was reproducibly induced with a 6 × 20 mm(2) annular lesion, with focal dysregulation of MMP gene expression, cell cloning in the inner AF, loss of NP aggrecan, and disc height. Loss of aggrecan from the NP was not attributable to increased proteolysis in the interglobular domain by MMPs or ADAMTS.


[Prevalence of microsporidia and other intestinal parasites in patients with HIV infection, Bogota, 2001].
Opportunistic intestinal parasites are a common cause of diarrhea in HIV-infected patients. To determine the prevalence of microsporidia and other opportunistic parasites infecting HIV patients in Bogotá, Colombia, 115 patients were examined for these infections during the year 2001. The institution and the sample percent from each are as follows: Santa Clara Hospital, 33.0%; San Pedro Claver, 20.0%; Simón Bolívar Hospital, 14.8%; San José Hospital, 13.9%; Central de la Policía Hospital, 6.1%; Compensar, 5.2%; Colombian League against AIDS, 2.6%; San Ignacio Hospital, 2.6%, and the Military Hospital, 1.7%. The average patient age was 36 years, with a range from 18 to 71 years. Patients with complaint of gastrointestinal symptoms were asked to provide two consecutive stool samples. The samples were concentrated in formalin-ether and examined microscopically for intestinal coccidian parasites by direct wet slide mounts. The prevalence of intestinal opportunistic parasites was 10.4% for Cryptosporidium sp. Initially, 29% of the samples were found to be positive for microsporidian spores using a modified Ziehl Neelsen chromotrope stain, but only 3.5% of them were confirmed as positive when a calcofluor/Gram chromotrope stain was used. The general prevalence of intestinal parasites was 59.1%. The most frequently found pathogens were Blastocystis hominis, 25.2%, and Entamoeba histolytica, 13%. In other studies with HIV patients in Colombia, lower prevalences of Cryptosporidium sp. infection were observed.


Lifestyle and metabolic factors for nonalcoholic fatty liver disease: Mendelian randomization study.
The risk factors for nonalcoholic fatty liver disease (NAFLD) have not been clearly identified. We conducted a Mendelian randomization (MR) study to explore this. Independent genetic variants strongly associated with 5 lifestyle and 9 metabolic factors were selected as instrumental variables from corresponding genome-wide association studies (GWASs). Summary-level data for NAFLD were obtained from a GWAS meta-analysis of 8434 cases and 770,180 non-cases (discovery dataset) and another GWAS meta-analysis of 1483 cases and 17,781 non-cases (replication dataset). Univariable and multivariable MR analyses were performed. There were associations with NAFLD for lifetime smoking index (odds ratio (OR) 1.59, 95% confidence interval (CI) 1.31-1.93 per SD-increase), body mass index (BMI, OR 1.33, 95% CI 1.23-1.43 per SD-increase), waist circumference (OR 1.82; 95% CI 1.48-2.24 per SD-increase), type 2 diabetes (OR 1.21, 95% CI 1.15-1.27 per unit increase in log-transformed odds), systolic blood pressure (OR 1.17; 95% CI 1.07-1.26 per 10 mmHg increase), high-density lipoprotein cholesterol (OR 0.84, 95% CI 0.77-0.90 per SD-increase), and triglycerides (OR 1.23, 95% CI 1.15-1.33 per SD-increase). The associations for type 2 diabetes, systolic blood pressure, triglycerides, but not for high-density lipoprotein cholesterol remained strong after adjusting for genetically-predicted BMI. Genetic liability to type 2 diabetes mediated 51.4% (95% CI 13.4-89.3%) of the BMI-effects on NAFLD risk. There were suggestive inverse associations of genetically-predicted alcohol, coffee, and caffeine consumption, and vigorous physical activity with NAFLD risk. This study identified several lifestyle and metabolic factors that may be causally implicated in NAFLD.


Hereditary Breast Cancer in the Brazilian State of Ceará (The CHANCE Cohort): Higher-Than-Expected Prevalence of Recurrent Germline Pathogenic Variants.
There is a significant lack of epidemiological data on hereditary cancer in Northeast Brazil. This is the largest study on the prevalence and mutational spectrum of cancer predisposition genes conducted in this region and the first in the State of Ceará. Patients ≥18 years of age that were referred to CHANCE (Grupo de Câncer Hereditário do Ceará) from March 2014 to December 2020 with testing criteria for breast cancer susceptibility genes according to NCCN v.1.2021 were eligible to participate. The inclusion of patients was limited to one individual per family and to those born in the State of Ceará. All patients underwent a hereditary cancer panel testing with at least 30 genes. A total of 355 patients were included, and 97 (27.3%) carried a P/LP germline variant in 18 different genes. Among the 97 P/LP carriers, BRCA1 (31, 31.9%) and BRCA2 (25, 25.7%) were the most frequently mutated genes, followed by PALB2 (10, 10.3%), CHEK2 (7, 7.2%) and ATM (4, 4.1%). A small number of recurrent variants (detected in three or more individuals) in BRCA1, BRCA2, CHEK2 and ATM represented the majority of the P/LP variants described in this cohort. In this cohort, the prevalence of L/PL was high, particularly involving the BRCA1, BRCA2, PALB2, CHEK2 and ATM genes and, to a lesser extent than expected, the TP53 gene. A high frequency of recurrent variants was also observed, for which further and larger analyses should clarify the presence of any possible founder effect. Characterizing the mutational profile of cancer predisposition genes in diverse populations may contribute to cancer prevention and therapeutic management.


A M23B family metallopeptidase of Helicobacter pylori required for cell shape, pole formation and virulence.
The molecular basis of the regulation of specific shapes and their role for the bacterial fitness remain largely unknown. We focused in this study on the Gram-negative and spiral-shaped Helicobacter pylori. To colonize its unique niche, H. pylori needs to reach quickly the human gastric mucosa, by swimming to and through the mucus layer. For that reason, the specific shape of H. pylori is predicted to be necessary for optimal motility in vivo, and consequently for its colonization ability. Here, we describe the involvement of a PG-modifying enzyme, HdpA (HP0506), in the mouse colonization ability of this bacterium, by regulating its shape. Indeed, the inactivation of the hp0506 gene led to a stocky and branched phenotype, affecting H. pylori colonization capacity despite a normal motility phenotype in vitro. In contrast, the overexpression of the hp0506 gene induced the transformation of H. pylori from rod to dividing cocci shaped bacteria. Furthermore, we demonstrated by PG analysis and enzymology, that HdpA carried both d,d-carboxypeptidase and d,d-endopeptidase activities. Thus, HdpA is the first enzyme belonging to the M23-peptidase family able to perform the d,d-carboxypeptidation and regulate cell shape.


Evaluation of Asteraceae herbal extracts in the management of diabetes and obesity. Contribution of caffeoylquinic acids on the inhibition of digestive enzymes activity and formation of advanced glycation end-products (in vitro).
The study was performed to assess, for the first time, the in vitro anti-diabetic potential of ten Asteraceae plant extracts to inhibit the activity of digestive enzymes (α-amylase, α-, β-glucosidases and lipase) responsible for hydrolysis/digestion of sugar and lipids. Prevention of advanced glycation end-products (AGEs) formation was evaluated in bovine serum albumin/ribose glycation reaction model. The phytochemical profiles and caffeoylquinic acids (CQAs) contents were determined for the methanolic extract of each plant. Analyzed plant extracts exhibited significant inhibitory activity against key digestive enzymes linked to type II diabetes and obesity. A strong inhibition was observed for glucosidases and mild activity towards amylase and lipase (compared to reference compounds). Moreover, some extracts exhibited potent ability to prevent formation of AGEs, implicated in some diabetic complications. Caffeoylquinic acids were dominant in all plant extracts and findings demonstrate that these compounds are the most relevant hypoglycemic and anti-glycation agents. From the obtained results, Argyranthemum pinnatifidum, Helichrysum melaleucum, and Phagnalon lowei are good candidates for further development of phyto-pharmaceutical preparations as complementary therapy for diabetes and obesity control.


TAM: a method for enrichment and depletion analysis of a microRNA category in a list of microRNAs.
MicroRNAs (miRNAs) are a class of important gene regulators. The number of identified miRNAs has been increasing dramatically in recent years. An emerging major challenge is the interpretation of the genome-scale miRNA datasets, including those derived from microarray and deep-sequencing. It is interesting and important to know the common rules or patterns behind a list of miRNAs, (i.e. the deregulated miRNAs resulted from an experiment of miRNA microarray or deep-sequencing). For the above purpose, this study presents a method and develops a tool (TAM) for annotations of meaningful human miRNAs categories. We first integrated miRNAs into various meaningful categories according to prior knowledge, such as miRNA family, miRNA cluster, miRNA function, miRNA associated diseases, and tissue specificity. Using TAM, given lists of miRNAs can be rapidly annotated and summarized according to the integrated miRNA categorical data. Moreover, given a list of miRNAs, TAM can be used to predict novel related miRNAs. Finally, we confirmed the usefulness and reliability of TAM by applying it to deregulated miRNAs in acute myocardial infarction (AMI) from two independent experiments. TAM can efficiently identify meaningful categories for given miRNAs. In addition, TAM can be used to identify novel miRNA biomarkers. TAM tool, source codes, and miRNA category data are freely available at http://cmbi.bjmu.edu.cn/tam.


Development of a phantom to evaluate the positioning accuracy of patient immobilization systems using thermoplastic mask and polyurethane cradle.
The purpose of this study was to develop a new phantom to evaluate the positioning accuracy of patient immobilization systems. The phantom was made of papers formed into a human shape, paper clay, and filling rigid polyester. Acrylonitrile butadiene styrene (ABS) pipes were inserted at anterior-posterior (A-P) and right-left (R-L) directions in the phantom to give static load by pulling ropes through the pipes. First, the positioning precision of the phantom utilizing a target locating system (TLS) was evaluated by moving the phantom on a couch along inferior-superior (I-S), A-P, and R-L directions in a range from -5 mm to +5 mm. The phantom's positions detected with the TLS were compared with values measured by a vernier caliper. Second, the phantom movements in a tensile test were chosen from patient movements determined from 15 patients treated for intracranial lesions and immobilized with a thermoplastic mask and polyurethane cradle. The phantom movement was given by minimum or maximum values of patient movements in each direction. Finally, the relationship between phantom movements and the static load in the tensile test was characterized from measurements using the new phantom and the TLS. The differences in all positions between the vernier caliper measurement and the TLS detected values were within 0.2 mm with frequencies of 100%, 95%, and 90% in I-S, A-P, and R-L directions, respectively. The phantom movements according to patient movements in clinical application in I-S, A-P, and R-L directions were within 0.58 mm, 0.94 mm, and 0.93 mm from the mean value plus standard deviation, respectively. The regression lines between the phantom movements and static load were given by y = 0.359x, y = 0.241x, and y = 0.451x in I-S, A-P, and R-L directions, respectively, where x is the phantom movement (mm) and y is the static load (kgf). The relationship between the phantom movements and static load may represent the performance of inhibiting patient movements, so the accuracy of the immobilization system in the intracranial lesion will be estimated in advance by basic tensile test on the new phantom. The newly developed phantom was useful to evaluate the accuracy of immobilization systems for a Cyberknife system for intracranial lesions.


Optimizing dendritic cell preparation for fusion with melanoma cells.
Fusion of dendritic cells (DCs) with melanoma cells could reinforce the antigenicity of tumors as a strategy for the treatment of malignant melanoma. However, the insufficient quantity of DCs and the low fusion efficiency limits the development of such approach. To define the dosage of the stimulating factors as well as the induction condition for the optimal DCs preparation and cell fusion. DCs were generated from murine bone marrow cells, and cultured with four different concentrations of the granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). DCs were confirmed to be mature by detecting the expression of MHC-II, CD11c, CD80, and CD83 by flow cytometry. DCs-melanoma fusion cells were generated using polyethylene glycols (PEG) with different molecular weights and the fusion efficiency was detected by fluorescence-activated cell sorter (FACS). The largest quantity of DCs was found when cells were cultured with 1000 U/ ml of GM-CSF and 500 U/ml of IL-4 (1.69 ± 0.04 ×10⁶ ml⁻¹, p<0.001 when compared with the other three groups). The expression levels of MHC-II and CD83 on day 7 after incubation were significantly lower than those on day 3 (MHC-II: p<0.001; CD83: p<0.001). The efficiency of cell fusion under induction of PEG-3000 was significantly higher than that of PEG-4000 (15.4 ± 0.56% vs. 11.1 ± 0.45%, p<0.001). The largest quantity for mature DCs was stimulated with 1000 U/ml of GM-CSF and 500 U/ml of IL-4 and the highest fusion efficiency was under induction of PEG-3000.


[A premature infant with increased bleeding tendency: what if the family history of bleeding disorders is negative?].
Haemophilia A is an X-chromosome recessive hereditary disorder and occurs in 1 in 5000 boys. In 30-50% of patients with haemophilia the family history of bleeding disorders is negative. We report on a premature male infant, born at 33 weeks of gestation, who exhibited prolonged bleeding from venipuncture sites the day after birth. The family history was negative for bleeding disorders. Initially he received vitamin K supplementation but, when the bleeding persisted, more detailed diagnostics revealed a prolonged aPTT. Factor VIII activity was 1% of the reference value and the neonate appeared to have a Grade I intraventricular haemorrhage on cerebral ultrasound. He was therefore treated with factor VIII. Early recognition and diagnostics in neonates with an increased bleeding tendency is important, even when the family history of bleeding disorders is negative.


An evaluation of the physiological demands of elite rugby union using Global Positioning System tracking software.
The current case study attempted to document the contemporary demands of elite rugby union. Players (n = 2) were tracked continuously during a competitive team selection game using Global Positioning System (GPS) software. Data revealed that players covered on average 6,953 m during play (83 minutes). Of this distance, 37% (2,800 m) was spent standing and walking, 27% (1,900 m) jogging, 10% (700 m) cruising, 14% (990 m) striding, 5% (320 m) high-intensity running, and 6% (420 m) sprinting. Greater running distances were observed for both players (6.7% back; 10% forward) in the second half of the game. Positional data revealed that the back performed a greater number of sprints (>20 km x h(-1)) than the forward (34 vs. 19) during the game. Conversely, the forward entered the lower speed zone (6-12 km x h(-1)) on a greater number of occasions than the back (315 vs. 229) but spent less time standing and walking (66.5 vs. 77.8%). Players were found to perform 87 moderate-intensity runs (>14 km x h(-1)) covering an average distance of 19.7 m (SD = 14.6). Average distances of 15.3 m (back) and 17.3 m (forward) were recorded for each sprint burst (>20 km x h(-1)), respectively. Players exercised at approximately 80 to 85% VO2max during the course of the game with a mean heart rate of 172 b x min(-1) ( approximately 88% HRmax). This corresponded to an estimated energy expenditure of 6.9 and 8.2 MJ, back and forward, respectively. The current study provides insight into the intense and physical nature of elite rugby using "on the field" assessment of physical exertion. Future use of this technology may help practitioners in design and implementation of individual position-specific training programs with appropriate management of player exercise load.


Morphology of the zoeal stages of Hexapus sexpes fabricius, 1798 (decapoda, brachyura, hexapodidae) reared in the laboratory.
Abstract: The present information is based on the study of the zoeal stages of Hexapus sexpes. A single ovigerous female of Hexapus sexpes was collected from the North West Persian Gulf (29 degrees 56'14"S, 49 degrees 0288"W) in April 2009. Female was maintained in aquarium containing natural sea water at 25 degrees C until the eggs hatched. The present study describes the zoeal stages of Hexapus sexpes. A complete description and illustration of larval features is provided and a comparison with closely related species (Spiroplax spiralis and Lambdophallus anfractus) showing known larval development is presented. The zoeal stages of H sexpes are very similar to that of L. anfractus. The zoeal of two species, however, could separate by the number of setae on the maxillule and maxilla.


Variations in osteoporosis medication utilization. A population-based ecological cross-sectional study in the region of Valencia, Spain.
Little is known about the contextual variability in osteoporosis medication utilization. Our aims were 1) to describe variations in utilization and spending on osteoporotic medication between the Primary Care Health Zones (PHZ) of the Valencia region, Spain, 2) to analyze observed variations using Small Area Variation Analysis methods, and 3) to quantify the influence of the specialized care level on variations in utilization. We conducted a population-based cross-sectional ecological study of expenditure and utilization of five therapeutic groups marketed as osteoporosis treatments in Spain in 2009. The unit of analysis was the PHZ (in total 240) nested in the 23 Hospital Healthcare Departments (HHD) of the region of Valencia, covering a population of about 4.9 million inhabitants. Drug utilization was measured by dispensed Defined Daily Dose per 1000 women aged 50 years old and over and day (DID) per PHZ and cost was measured by the annual osteoporosis drug cost per woman aged 50 and older as well as the average price of DDD (Defined Daily Dose) in each PHZ. We calculated Indirect Standardized Drug Utilization Ratios (ISR) and we used Spearman's correlation to analyze associations between the ISRs of the different therapies. The average osteoporosis drug consumption was 119.1 DID, ranging from 77.6 to 171.3 DID (2.2 times higher) between PHZs in the 5th and 95th percentiles. Annual expenditure also showed a two-fold variation among PHZs. Average prices of the DDD by therapeutic group showed very low or no variation, although they differed substantially among therapeutic groups. Regarding the standardized consumption of osteoporotic drugs, HHDs explained a substantial part (39%) of the variance among PHZs. In conclusion, there is considerable variability in the volume and choice of anti-osteoporotic treatments between PHZs. with HHDs explaining an important proportion of the variation in utilization. Interventions aimed at reducing variation to improve appropriate care should take into account both the PHZ and HHD levels of care.


Physical Fatigue, Fitness, and Muscle Function in Patients With Antineutrophil Cytoplasmic Antibody-Associated Vasculitis.
This study investigated differences in cardiorespiratory fitness, muscular function, perceived exertion, and anxiety/depression between patients and healthy controls (HCs) and assessed which of these variables may account for the fatigue experienced by patients. Fatigue was measured in 48 antineutrophil cytoplasmic antibody-associated vasculitis patients and 41 healthy controls using the Multidimensional Fatigue Inventory (MFI-20), focusing on the physical component. Quality of life, anxiety/depression, and sleep quality were assessed by validated questionnaires. Muscle mass was measured by dual-energy x-ray absorptiometry scan, strength as the maximal voluntary contraction (MVC) force, and endurance as sustained isometric contraction at 50% MVC of the quadriceps. Voluntary activation was assessed by superimposed electrical stimulation. Cardiorespiratory fitness ( ˙Vo2 max and oxygen pulse [O2 pulse]) and perceived exertion (Borg scale) were measured during progressive submaximal exercise. Patients reported elevated physical fatigue scores compared to HCs (patients MFI-20 physical 13 [interquartile range (IQR) 8-16], HCs MFI-20 physical 5.5 [IQR 4-8]; P < 0.001). Muscle mass was the same in both groups, but MVC and time to failure in the endurance test were lower due to reduced voluntary activation in patients. Estimated ˙Vo2 max and O2 pulse were the same in both groups. For the same relative workload, patients reported higher ratings of perceived exertion, which correlated with reports of MFI-20 physical fatigue (R(2) = 0.2). Depression (R(2) = 0.6), anxiety (R(2) = 0.3), and sleep disturbance (R(2) = 0.3) were all correlated with MFI-20 physical fatigue. These observations suggest that fatigue in patients is of a central rather than peripheral origin, supported by associations of fatigue with heightened perception of exertion, depression, anxiety, and sleep disturbance but normal muscle and cardiorespiratory function.


Mast cells in the pinna of Balb/c 'nude' (nu/nu) and heterozygotes (ny/+)mice.
The relative number of mast cells in the ear lobes' skin (pinna) of nude (athymic) nu/nu and normal (thymic) nu/+ heterozygotes of Balb/c mice was similar. The results obtained contradict some suggestions about the general influence of the thymus on the number of mast cells in the skin and suggest the existence of some local factor(s) in regulation of skin mast cell numbers.


Protein structure modelling and evaluation based on a 4-distance description of side-chain interactions.
Accurate evaluation and modelling of residue-residue interactions within and between proteins is a key aspect of computational structure prediction including homology modelling, protein-protein docking, refinement of low-resolution structures, and computational protein design. Here we introduce a method for accurate protein structure modelling and evaluation based on a novel 4-distance description of residue-residue interaction geometry. Statistical 4-distance preferences were extracted from high-resolution protein structures and were used as a basis for a knowledge-based potential, called Hunter. We demonstrate that 4-distance description of side chain interactions can be used reliably to discriminate the native structure from a set of decoys. Hunter ranked the native structure as the top one in 217 out of 220 high-resolution decoy sets, in 25 out of 28 "Decoys 'R' Us" decoy sets and in 24 out of 27 high-resolution CASP7/8 decoy sets. The same concept was applied to side chain modelling in protein structures. On a set of very high-resolution protein structures the average RMSD was 1.47 A for all residues and 0.73 A for buried residues, which is in the range of attainable accuracy for a model. Finally, we show that Hunter performs as good or better than other top methods in homology modelling based on results from the CASP7 experiment. The supporting web site http://bioinfo.weizmann.ac.il/hunter/ was developed to enable the use of Hunter and for visualization and interactive exploration of 4-distance distributions. Our results suggest that Hunter can be used as a tool for evaluation and for accurate modelling of residue-residue interactions in protein structures. The same methodology is applicable to other areas involving high-resolution modelling of biomolecules.


The Royal Flying Doctor Service Flies to New Heights: The Journey of Health Information Management.
The Royal Flying Doctor Service (RFDS) of Australia was founded in 1928 by the Reverend John Flynn to deliver health services to the people of the Australian Outback. In this unique environment the RFDS Queensland Section provides both Primary Health Care and Aeromedical services to rural and remote communities throughout Queensland. It provides health services from a hub and spoke model and its clinicians work very closely with other health service providers, such as Queensland Heath, within the communities it visits. Currently, the RFDS' health records are both paper and electronic and clinicians duplicate much of patient information and data between RFDS and non-RFDS health records. Introduction of an off-the-shelf electronic medical record (EMR) would not meet the RFDS' clinical and organisational needs because of complexity, the multidisciplinary nature of the teams and the lack of communication technology in the communities the RFDS visits. This article defines the vision for a health information system designed to meet the requirements of the RFDS, and describes its implementation throughout RFDS Queensland using the PRINCE2 project management methodology.


Current Landscape and Future Prospects of Radiation Sensitizers for Malignant Brain Tumors: A Systematic Review.
Radiation therapy (RT) is the cornerstone of management of malignant brain tumors, but its efficacy is limited in hypoxic tumors. Although numerous radiosensitizer compounds have been developed to enhance the effect of RT, progress has been stagnant. Through this systematic review, we provide an overview of radiosensitizers developed for malignant brain tumors, summarize their safety and efficacy, and evaluate areas for possible improvement. Following PRISMA guidelines, PubMed, EMBASE, Cochrane, and Web of Science were searched using terminology pertaining to radiosensitizers for brain tumor RT. Articles reporting clinical evidence of nonantineoplastic radiosensitizers with RT for malignant central nervous system tumors were included. Data of interest were presumed mechanism of action, median overall survival (OS), progression-free survival (PFS), and adverse events. Twenty-two unique radiosensitizers were identified. Only 2/22 agents (fluosol with oxygen, and efaproxiral) showed improvement in OS in patients with glioblastoma and brain metastasis, respectively. A larger study was not able to confirm the latter. Improved PFS was reported with use of metronidazole, sodium glycididazole, and chloroquine. There was a wide range of toxicities, which prompted change of schedule or complete discontinuation of 9 agents. Progress in radiosensitizers for malignant CNS tumors has been limited. Only 2 radiosensitizers have shown limited improvement in survival. Alternative strategies such as synthetic drug design, based on a mechanism of action that is independent of crossing the blood-brain barrier, may be necessary. Use of drug development strategies using new technologies to overcome past challenges is necessary.


Discovery of protein-RNA networks.
Coding and non-coding RNAs associate with proteins to perform important functions in the cell. Protein-RNA complexes are essential components of the ribosomal and spliceosomal machinery; they are involved in epigenetic regulation and form non-membrane-bound aggregates known as granules. Despite the functional importance of ribonucleoprotein interactions, the precise mechanisms of macromolecular recognition are still poorly understood. Here, we present the latest developments in experimental and computational investigation of protein-RNA interactions. We compare performances of different algorithms and discuss how predictive models allow the large-scale investigation of ribonucleoprotein associations. Specifically, we focus on approaches to decipher mechanisms regulating the activity of transcripts in protein networks. Finally, the catRAPID omics express method is introduced for the analysis of protein-RNA expression networks.


Dosimetric consequences of misalignment and realignment in prostate 3DCRT using intramodality ultrasound image guidance.
It is common practice to correct for interfraction motion by shifting the patient from reference skin marks to better align the internal target at the linear accelerator's isocenter. Shifting the patient away from skin mark alignment causes the radiation beams to pass through a patient geometry different from that planned. Yet, dose calculations on the new geometry are not commonly performed. The intention of this work was to compare the dosimetric consequences of treating the patient with and without setup correction for the common clinical scenario of prostate interfraction motion. In order to account for prostate motion, 32 patients initially aligned to the room lasers via skin marks were realigned under the treatment beams by shifting the treatment couch based on ultrasound image guidance. An intramodality 3D ultrasound image guidance system was used to determine the setup correction, so that errors stemming from different tissue representations on different imaging modalities were eliminated. Two scenarios were compared to the reference static treatment plan: (1) Uncorrected patient alignment and (2) corrected patient alignment. Prostate displacement statistics and the dose to the clinical target volume (CTV), bladder, and rectum are reported. Monte Carlo dose calculation methods were employed. Comparing the uncorrected and corrected scenarios using the static treatment plan as the reference, the average percent difference in D95 for the CTV improved from -5.1% (range -40%, 1.3%) to 0.0% (-3.5%, 2.0%) and the average percent difference in V90 for the bladder and rectum changed from -11% (-84%, 232%) to -8.3% (-61%, 5.2%) and from -47% (-100%, 108%) to 0.9% (-62%, 102%), respectively. There was no simple correlation between displacement and dose discrepancy before correction. After patient realignment, the prescribed dose to the CTV was achieved within 1% for 75% (24/32) of the patients. After patient realignment, 50% of the patients had doses that differed from the static treatment plan by 25% for the bladder and 8% for the rectum. The dose degradation due to prostate motion (before correction) is not accurately predicted from the average trends for all patients. Outliers included smaller displacements that lead to larger dosimetric differences in the corrected scenario, especially for the bladder and rectum, which exhibited doses substantially different from that planned.


Fabrication of ultra-dense sub-10 nm in-plane Si nanowire arrays by using a novel block copolymer method: optical properties.
The use of a low-χ, symmetric block copolymer as an alternative to the high-χ systems currently being translated towards industrial silicon chip manufacture has been demonstrated. Here, the methodology for generating on-chip, etch resistant masks and subsequent pattern transfer to the substrate using ultra-small dimension, lamellar, microphase separated polystyrene-b-poly(ethylene oxide) (PS-b-PEO) block copolymer (BCP) is described. Well-controlled films of a perpendicularly oriented lamellar pattern with a domain size of ∼8 nm were achieved through amplification of an effective interaction parameter (χeff) of the BCP system. The self-assembled films were used as 'templates' for the generation of inorganic oxides nanowire arrays through selective metal ion inclusion and subsequent processing. Inclusion is a significant challenge because the lamellar systems have less chemical and mechanical robustness than the cylinder forming materials. The oxide nanowires of uniform diameter (∼8 nm) were isolated and their structure mimics the original BCP nanopatterns. We demonstrate that these lamellar phase iron oxide nanowire arrays could be used as a resist mask to fabricate densely packed, identical ordered, good fidelity silicon nanowire arrays on the substrate. Possible applications of the materials prepared are discussed, in particular, in the area of photonics and photoluminescence where the properties are found to be similar to those of surface-oxidized silicon nanocrystals and porous silicon.


[Magnetic resonance imaging (MRI) in the diagnosis of recurrences of ovarian cancer in the small pelvis].
The paper provides the results of small pelvic magnetic resonance tomography (MRI) in 62 patients with ovarian cancer after primary special treatment. Out of them 50 patients were found to have recurrences and metastases of the underlying disease, 12 patients had clinical remission. The study yielded MR signs and MR semiotics of recurrences of ovarian cancer in the small pelvis. The capacities of MRI with low and high intensities of a magnetic field were comparatively studied in the diagnosis of recurrences and metastases of ovarian cancer.


[Noonan syndrome: from phenotype to growth hormone therapy].
Noonan Syndrome (NS) is one of the most common genetic syndromes and it is an important differential diagnosis in children with short stature, delayed puberty and cryptorchidism. NS is characterized by dysmorphic facial features, congenital heart defects and short stature, but there is a great variability in phenotype. NS may occur in a pattern consistent with autosomal dominant inheritance with almost complete penetrance. The diagnosis is based on a clinical score system proposed by van der Burgt e cols. in 1994. In recent years, germline mutations in the components of RAS-MAPK (mitogen activated protein kinase) pathway have been shown to be involved in the pathogenesis of NS. Mutations in PTPN11, KRAS, SOS1, RAF1 and MEK1 can explain 60-70% of NS molecular cause. Growth hormone therapy is proposed to correct the short stature observed in these patients. Recent studies suggest that the presence of PTPN11 mutations in patients with NS indicates a reduced growth response to short-term hrGH treatment. In this article, it is reviewed clinical and molecular aspects of NS and hrGH treatment for short stature.


Inference of gene regulatory networks incorporating multi-source biological knowledge via a state space model with L1 regularization.
Comprehensive understanding of gene regulatory networks (GRNs) is a major challenge in the field of systems biology. Currently, there are two main approaches in GRN analysis using time-course observation data, namely an ordinary differential equation (ODE)-based approach and a statistical model-based approach. The ODE-based approach can generate complex dynamics of GRNs according to biologically validated nonlinear models. However, it cannot be applied to ten or more genes to simultaneously estimate system dynamics and regulatory relationships due to the computational difficulties. The statistical model-based approach uses highly abstract models to simply describe biological systems and to infer relationships among several hundreds of genes from the data. However, the high abstraction generates false regulations that are not permitted biologically. Thus, when dealing with several tens of genes of which the relationships are partially known, a method that can infer regulatory relationships based on a model with low abstraction and that can emulate the dynamics of ODE-based models while incorporating prior knowledge is urgently required. To accomplish this, we propose a method for inference of GRNs using a state space representation of a vector auto-regressive (VAR) model with L1 regularization. This method can estimate the dynamic behavior of genes based on linear time-series modeling constructed from an ODE-based model and can infer the regulatory structure among several tens of genes maximizing prediction ability for the observational data. Furthermore, the method is capable of incorporating various types of existing biological knowledge, e.g., drug kinetics and literature-recorded pathways. The effectiveness of the proposed method is shown through a comparison of simulation studies with several previous methods. For an application example, we evaluated mRNA expression profiles over time upon corticosteroid stimulation in rats, thus incorporating corticosteroid kinetics/dynamics, literature-recorded pathways and transcription factor (TF) information.


Biomarkers of environmental contaminants in the coastal waters of Estonia (Baltic Sea): effects on eelpouts (Zoarces viviparus).
The eastern Baltic Sea near the Estonian coast is heavily navigated by numerous cargo ships and oil tankers. Hundreds of accidents and oil spills happen yearly in this area. Yet, there is a lack of data concerning the distribution and effects of the environmental contaminants, especially polycyclic aromatic hydrocarbons (PAHs). Different parts of the Baltic Sea have different levels of contamination; therefore a wide range of monitoring stations in coastal areas in the Gulf of Finland and Gulf of Riga were chosen. The aim of the present research was to document the responses of chosen biomarkers of environmental contaminants in different unstudied areas of the Estonian coastal sea. During 2009 and 2010 we measured PAH metabolites, EROD activities, geno- and cytotoxicity, histology, parasites and other biomarkers from the eelpout (Zoarces viviparus), a resident benthic fish species. The results showed that fish from the Gulf of Riga emitted lower levels of fluorescence in fixed wavelength analyses (representing equivalents of PAH metabolites in bile and urine), and consistently, showed less geno- and cytotoxicity and parasite infection, higher liver somatic index (LSI) and a higher condition factor (CF) than fish inhabiting areas close to the Baltic proper and in the Gulf of Finland. The results point to the effect of long-range contaminant transportation, whether atmospheric or hydrodynamic, and also to the intensive shipping activity in international routes. This study fills the gap of knowledge in this area that has persisted until now. Nevertheless, more studies in this area on the different groups of contaminants are necessary, to specify the factors that are responsible for observed biological effects.


A versatile pH-responsive platform for intracellular protein delivery using calcium phosphate nanoparticles.
Intracellular protein delivery has great biomedical applications; the safe and efficient delivery vectors are crucial for achieving this goal. Here we report a platform for efficient protein delivery using calcium phosphate (CaP) nanoparticles. The well-dispersed and highly-stable nanoparticles (∼100 nm) are prepared with the protein loading capacity of up to 29%. The nanoparticles are stable under serum conditions; however, after being internalized into cells, the particles quickly release protein in the weak acidic endosomes/lysosomes. The decomposition of CaP promotes the endo-lysosomal escape of proteins released from nanoparticles. The protein/CaP conjugates were prepared under mild conditions (aqueous solution, room temperature); hence the protein released from nanoparticles retained its folding and function. In addition, all materials used for the preparation are highly biocompatible. This method has been applied for the loading of three model proteins, BSA, GFP and KillerRed; similar loading properties were observed on these proteins. Therefore, this work offers a general approach for intracellular protein delivery, which could be applicable for therapeutic proteins.


Eccentric Contraction Enhances Healing of the Bone-Tendon Interface After Rotator Cuff Repair in Mice.
Various muscle contraction modalities have differing effects on the musculoskeletal system. To understand the magnitude of these effects, the authors investigated the effects of eccentric and concentric contractions on the bone-tendon interface after rotator cuff repair in mice. Eccentric contraction promotes healing of the bone-tendon interface after rotator cuff repair in mice better than other muscle contraction patterns. Controlled laboratory study. The authors performed acute supraspinatus tendon repair of the right shoulder in 104 C57BL/6 mice. Animals were randomized into 4 groups postoperatively: control group (Con group), horizontal running group (Horz group), +15° uphill running group (Up group), and -15° downhill running group (Down group), with 26 animals in each group. At 4 and 8 weeks postoperatively, the authors removed the eyeball, collected blood samples, and extracted the supraspinatus tendon-humerus complex for histological, immunological, bone morphological, and biomechanical tests. At 4 and 8 weeks postoperatively, the Down group exhibited a better collagen cell arrangement and fibrocartilage layer than the other 3 groups. At 4 weeks postoperatively, anti-inflammatory macrophages (M2 macrophages) were observed at the repair site in all groups except for the Con group. At 8 weeks postoperatively, M2 macrophages were withdrawn from the tendon site in all groups. The transforming growth factor β1 concentration in the Down group was greater than that in the other 3 groups at 4 weeks postoperatively, and it was higher than that in the Con group at 8 weeks postoperatively. The bone volume fraction, number of trabeculae, and thickness of trabeculae at the repair site in the Down group, as well as the ultimate strength and failure load in the biomechanical tests, were greater than those in the other 3 groups at 8 weeks postoperatively. Eccentric contraction promotes healing of the bone-tendon interface after rotator cuff repair in mice better than other muscle contraction patterns. After clinical rotator cuff repair, patients can be rehabilitated by eccentric training to speed up the functional recovery of the shoulder joint.


Microwave Properties of One-dimensional Photonic Structures Based on Composite Layers Filled with Nanocarbon.
This work presents the results of computer modeling and experimental measurements of microwave transmission properties for one-dimensional periodic multi-layered photonic structures (PCs), composed of epoxy layers and composite layers filled with nanocarbon particles-multi-walled carbon nanotubes and graphite nanoplatelets. The results show that the characteristics of observed photonic band gaps in transmission spectra of PC can be controlled by varying the parameters of layers, namely, the complex permittivity and the layer thickness. It was found that the insertion of the defects (for instance, magnetic layer) into photonic structure can change the EMR transmission spectrum. The comparative analysis of EMR transmission spectra for investigated photonic structures has showed good agreement between the experimental and simulated data. It was found that EMR absorption in composite layers of photonic structures shifts the transmission spectra to the smaller values of EMR transmission index and reduces the sharpness of photonic band gaps. Thus, by changing the parameters of composite layers in photonic structure, we can obtain the tunable photonic band gaps, necessary for technological applications in devices, capable of storing, guiding, and filtering microwaves.


Acetaminophen-induced hepatic neutrophil accumulation and inflammatory liver injury in CD18-deficient mice.
Acetaminophen (APAP) hepatotoxicity is currently the most frequent cause of acute liver failure in the US and many European countries. Although intracellular signalling mechanisms are critical for hepatocellular injury, a contribution of inflammatory cells, especially neutrophils, has been suggested. However, conflicting results were obtained when using immunological intervention strategies. The role of neutrophils was investigated using a CD18-deficient mouse model. Treatment of C57Bl/6 wild type mice with 300 mg/kg APAP resulted in severe liver cell necrosis at 12 and 24 h. This injury was accompanied by formation of cytokines and chemokines and accumulation of neutrophils in the liver. However, there was no difference in the inflammatory response or liver injury in CD18-deficient mice compared with wild-type animals. In contrast to treatment with endotoxin, no upregulation of CD11b or priming for reactive oxygen was observed on neutrophils isolated from the peripheral blood or the liver after APAP administration. Furthermore, animals treated with endotoxin 3 h after APAP experienced an exaggerated inflammatory response as indicated by substantially higher cytokine and chemokine formation and twice the number of neutrophils in the liver. However, liver injury in the two-hit model was the same as with APAP alone. Our data do not support the hypothesis that neutrophils contribute to APAP hepatotoxicity or that a neutrophil-mediated injury phase could be provoked by a second, pro-inflammatory hit. Thus, APAP-induced liver injury in mice is dominated by intracellular mechanisms of cell death rather than by neutrophilic inflammation.


Differential Histone Acetylation in Sub-Regions of Bed Nucleus of the Stria Terminalis Underlies Fear Consolidation and Extinction.
The hallmark of anxiety disorders is excessive fear. Previous studies have suggested that selective neural projections from Basal nucleus of stria terminalis (BNST) to amygdala and vice-versa precisely control the fear learning process. However the exact mechanism how the BNST controls fear consolidation and its extinction is largely unknown. In the present study we observed the changes in the BNST sub-regions following fear conditioning and its extinction. The change in the number of positive neurons was determined by immunohistochemistry for Acetyl H3 (Histone 3), Acetyl H4 (Histone 4), cAMP response element binding Protein (CBP) and c-fos in three sub-regions of the BNST namely the anterio-lateral BNST (STLP) and anterio-medial BNST (STMA), and lateral-ventral BNST (STLV) of rats subjected to auditory fear conditioning and extinction. We found significant increase in the number of CBP, acetyl H3 and acetyl H4 positive neurons in the STMA and STLV but not in the STLP after fear conditioning. However, following fear extinction the number of CBP, acetyl H3 and acetyl H4 positive neurons increased significantly in the STLP but not in the STMA and STLV. Similar changes were observed in the number of c-fos positive neurons after fear consolidation and extinction. The results from this study suggest that the differential histone acetylation in the different sub-regions of the BNST following fear learning and its extinction may be responsible for changes in the neuronal activation patterns resulting in either fear or less fear.


Interpersonal relations between health care workers and young clients: barriers to accessing sexual and reproductive health care.
Interpersonal relations between health care providers and young clients have long being cited as an important element for improving client up take of services, satisfaction and overall health outcomes. In an era of HIV and AIDS this forms a critical determinant to young people accessing sexual and reproductive health care. This study explores to what extent interpersonal relations form a barrier to young peoples access to and satisfaction of health services. The study draws on data from 200 client exit interviews and four in-depth interviews conducted with university students and university health care staff in Kwazulu-Natal, South Africa. While young people are aware of the importance of utilising STI, HIV and family planning services they experienced barriers in their relationship with providers. This served as a deterrent to their use of the health facility. Adequate training in interpersonal relations for youth-friendly service provision is essential in helping overcome communication problems and enabling providers to interact with young clients at a more personal level.


[Metabolic changes in arachidonic acid during aspirin desensitization].
Aspirin sensitivity occurs in 10% of all asthmatics patients. In this subset of asthmatics, nasal congestion and bronchospasm occurs between 30-180 minutes after ingestion of aspirin. Following a respiratory reaction to aspirin, all patients can be desensitized to aspirin by repetitively introducing small and then larger doses of aspirin until the asthmatic subject can ingest 650 mg of aspirin without adverse effect. The mechanism of aspirin sensitivity are incompletely understood. And the reasons why ASA desensitization occurs universally are unknown. In this study, known ASA sensitive and control insensitive asthmatics were challenged with ASA. Urine was collected before, during induced bronchospasm, and after ingestion of 650 mg of ASA when the adverse effect (ie., acute desensitization) had subsided. Excretion levels of cyclo-oxygenase and lipoxygenase products in the urine were determined.


Determination of metabolic activity in planktonic and biofilm cells of Mycoplasma fermentans and Mycoplasma pneumoniae by nuclear magnetic resonance.
Mycoplasmas are fastidious microorganisms, typically characterised by their restricted metabolism and minimalist genome. Although there is reported evidence that some mycoplasmas can develop biofilms little is known about any differences in metabolism in these organisms between the growth states. A systematic metabolomics approach may help clarify differences associated between planktonic and biofilm associated mycoplasmas. In the current study, the metabolomics of two different mycoplasmas of clinical importance (Mycoplasma pneumoniae and Mycoplasma fermentans) were examined using a novel approach involving nuclear magnetic resonance spectroscopy and principle component analysis. Characterisation of metabolic changes was facilitated through the generation of high-density metabolite data and diffusion-ordered spectroscopy that provided the size and structural information of the molecules under examination. This enabled the discrimination between biofilms and planktonic states for the metabolomic profiles of both organisms. This work identified clear biofilm/planktonic differences in metabolite composition for both clinical mycoplasmas and the outcomes serve to establish a baseline understanding of the changes in metabolism observed in these pathogens in their different growth states. This may offer insight into how these organisms are capable of exploiting and persisting in different niches and so facilitate their survival in the clinical setting.


Exosomal microRNAs from Longitudinal Liquid Biopsies for the Prediction of Response to Induction Chemotherapy in High-Risk Neuroblastoma Patients: A Proof of Concept SIOPEN Study.
Despite intensive treatment, 50% of children with high-risk neuroblastoma (HR-NB) succumb to their disease. Progression through current trials evaluating the efficacy of new treatments for children with HR disease usually depends on an inadequate response to induction chemotherapy, assessed using imaging modalities. In this study, we sought to identify circulating biomarkers that might be detected in a simple blood sample to predict patient response to induction chemotherapy. Since exosomes released by tumor cells can drive tumor growth and chemoresistance, we tested the hypothesis that exosomal microRNA (exo-miRNAs) in blood might predict response to induction chemotherapy. The exo-miRNAs expression profile in plasma samples collected from children treated in HR-NBL-1/SIOPEN before and after induction chemotherapy was compared to identify a three exo-miRs signature that could discriminate between poor and good responders. Exo-miRNAs expression also provided a chemoresistance index predicting the good or poor prognosis of HR-NB patients.


Intraoperative detection of liver tumors aided by a fluorescence goggle system and multimodal imaging.
Real-time image guidance in the operating room is needed to improve instantaneous surgical decisions. Toward this goal, we utilized a new fluorescence goggle system and a near-infrared fluorescent dye approved for human use, indocyanine green, to demonstrate the feasibility of detecting liver tumors intraoperatively. The fluorescence goggle provided successful imaging of multifocal breast cancer metastases in mouse liver. Diffused tumor deposits as small as 0.8 mm in diameter were detected, which were not obvious without the fluorescence goggle. A combination of surface-weighted fluorescence imaging and deep tissue-sensitive ultrasound imaging allowed comprehensive image guidance with the fluorescence goggle system for tumor resection in a rabbit VX2 liver metastasis model. This multimodal detection and guided surgical intervention strategy using ultrasonic imaging and real-time intraoperative fluorescence guidance is a promising and innovative technology platform for improving surgical outcome of human patients with primary or metastatic liver cancer.


Evaluation of intestinal metabolism and absorption using the Ussing chamber system equipped with intestinal tissue from rats and dogs.
The purpose of this study was to evaluate the intestinal metabolism and absorption in a mini-Ussing chamber equipped with animal intestinal tissues, based on the transport index (TI). TI value was defined as the sum of drug amounts transported to the basal-side component (Xcorr) and drug amounts accumulated in the tissue (Tcorr), which are normalized by AUC of a drug in the apical compartment, as an index for drug absorption. Midazolam was used as a test compound for the evaluation of intestinal metabolism and absorption. The metabolite formulation of midazolam was observed in both rats and dogs. Ketoconazole inhibited the intestinal metabolism of midazolam in rats and improved its intestinal absorption to a statistically significant extent. Therefore, the mini-Ussing chamber, equipped with animal intestinal tissues, showed potential to use the evaluation of the intestinal metabolism and absorption, including the assessment of species differences.


Melatonin and its potential biological functions in the fruits of sweet cherry.
Melatonin is a well-known molecule which possesses many beneficial effects on human health. Many agriculture products provide natural melatonin in the diet. Cherry is one such fruit as they are rich in melatonin. In order to understand the biological roles of melatonin in cherry fruit, melatonin synthesis and its changes over 24 hr period were systematically monitored both during their development and in the ripe cherries in two cultivars, 'Hongdeng' (Prunus avium L. cv. Hongdeng) and 'Rainier' (Prunus avium L. cv. Rainier). It was found that both darkness and oxidative stress induced melatonin synthesis, which led to dual melatonin synthetic peaks during a 24 hr period. The high levels of malondialdehyde induced by high temperature and high intensity light exposure were directly related to up-regulated melatonin production. A primary function of melatonin in cherry fruits is speculated to be as an antioxidant to protect the cherry from the oxidative stress. Importantly, plant tryptophan decaboxylase gene (PaTDC) was identified in cherry fruits. Our data shows that PaTDC expression is positively related to the melatonin production in the cherry. This provides additional information to suggest that tryptophan decaboxylase is a rate-limiting enzyme of melatonin synthesis in plants.


Detection of 6-(methylsulfinyl)hexyl isothiocyanate (6-MITC) and its conjugate with N-acetyl-L-cysteine (NAC) by high performance liquid chromatograpy-atmospheric pressure chemical ionization mass spectrometry (HPLC-MS/APCI).
A method using high-performance liquid chromatography (HPLC) and atmospheric pressure chemical ionization (APCI) mass spectrometry (MS) was established for the detection of 6-(methylsulfinyl)hexyl isothiocyanate (6-MITC) and its conjugate with N-acetyl-L-cysteine (NAC). The optimal chromatographic conditions were obtained on an ODS column (150 x 4.6 mm, 3 microm) with the column temperature at 37 degrees C. The mobile phase consisted of a methanol-0.1% trifluoroacetic acid (TFA) mixture (50 : 50, v/v), and the flow rate was 0.3 ml/min. The detection wavelength was set at 220 nm. The identities of the peaks were accomplished by comparing retention times (tR), UV and mass data. All calibration curves showed good linear regression (correlation coefficients for 6-MITC and NAC>0.999) within test ranges. The developed method provided satisfactory precision calculated as percent coefficient of variation with overall intra-day and inter-day variations of less than 5% (4.1 and 4.9% for 6-MITC; 4.2 and 4.9% for NAC). Both 6-MITC and NAC had good responses in the positive APCI and formed strong [M+H]+ ions in the full scan spectra at an m/z of 206 and 164, respectively. The presence of the [M+H]+ ion for the 6-MITC/NAC conjugate was also observed at an m/z of 369. To our best knowledge, this is the first report that describes the formation of the 6-MITC/NAC conjugate and its detection method by HPLC-MS.