Long-term motor development after hypothermia-treated hypoxic-ischaemic encephalopathy.
To describe longitudinal motor development in children treated with therapeutic- hypothermia (TH) due to neonatal hypoxic-ischaemic encephalopathy (HIE) and to explore motor functioning in early adolescence. Children treated with TH due to HIE during 2007-2009, in Stockholm, participated in a prospective follow-up study. Motor development was assessed on four occasions, reported as percentiles and at mean ages. Alberta Infant Motor Scale was used at 0.35 years of age, Bayley Scales of Infant and Toddler Development-III at 2.1 years and Movement Assessment Battery for Children (MABC-2) at 7.3 and 11.1 years of age. MABC-2 Checklist was completed by parents at 7.3 and 11.1 years of age. General cognition was assessed using Wechsler Intelligence Scales for Children Fifth Edition (WISC-V). Thirty-one percent (14/45) of the children had a motor score ≤ 15th percentile, indicating risk of motor difficulties at 11.1 years of age, and simultaneously the scores from parents of 52% (23/44), indicating risk of motor difficulties in the everyday context. These children had significantly lower motor percentile at 2.1 years of age, but within the normal range. Longitudinal motor development displayed a weak association with WISC-V Full Scale IQ (rs0.38, p = 0.013). Among survivors of hypothermia-treated HIE free of moderate/severe cerebral palsy, a third had MABC-2 scores indicating risk of motor difficulties at 11.1 years of age. As motor difficulties became more apparent over time, we suggest that children treated with TH due to neonatal HIE should be followed into at least middle school age.


RNA-Seq analysis uncovers transcriptomic variations between morphologically similar in vivo- and in vitro-derived bovine blastocysts.
A valuable tool for both research and industry, in vitro fertilization (IVF) has applications range from gamete selection and preservation of traits to cloning. Although IVF has achieved worldwide use, with approximately 339,685 bovine embryos transferred in 2010 alone, there are still continuing difficulties with efficiency. It is rare to have more than 40% of fertilized in vitro cattle oocytes reach blastocyst stage by day 8 of culture, and pregnancy rates are reported as less than 45% for in vitro produced embryos. To investigate potential influences in-vitro fertilization (IVF) has on embryonic development, this study compares in vivo- and in vitro-derived bovine blastocysts at a similar stage and quality grade (expanded, excellent quality) to determine the degree of transcriptomic variation beyond morphology using RNA-Seq. A total of 26,906,451 and 38,184,547 fragments were sequenced for in vitro and in vivo embryo pools, respectively. We detected expression for a total of 17,634 genes, with 793 genes showing differential expression between the two embryo populations with false discovery rate (FDR) < 0.05. There were also 395 novel transcribed units found, of which 45 were differentially expressed (FDR < 0.05). In addition, 4,800 genes showed evidence of alternative splicing, with 873 genes displaying differential alternative splicing between the two pools (FDR < 0.05). Using GO enrichment analysis, multiple biological pathways were found to be significantly enriched for differentially expressed genes (FDR < 0.01), including cholesterol and sterol synthesis, system development, and cell differentiation. Thus, our results support that IVF may influence at the transcriptomic level and that morphology is limited in full characterization of bovine preimplantation embryos.


Cardiac and lung complication probabilities after breast cancer irradiation.
To assess for locoregional irradiation of breast cancer patients, the dependence of cardiac (cardiac mortality) and lung (radiation pneumonitis) complications on treatment technique and individual patient anatomy. Three-dimensional treatment planning was performed for 30 patients with left-sided breast cancer and various breast sizes. Two locoregional techniques (Techniques A and B) and a tangential field technique, including only the breast in the target volume, were planned and evaluated for each patient. In both locoregional techniques tangential photon fields were used to irradiate the breast. The internal mammary (IM)-medial supraclavicular (MS) lymph nodes were treated with an anterior mixed electron/photon field (Technique A) or with an obliquely incident mixed electron/photon IM field and an anterior electron/photon MS field (Technique B). The optimal IM and MS electron field dimensions and energies were chosen on the basis of the IM-MS lymph node target volume as delineated on CT-slices. The position of the tangential fields was adapted to match the IM-MS fields. Dose-volume histograms (DVHs) and normal tissue complication probabilities (NTCPs) for the heart and lung were compared for the three techniques. In the beam's eye view of the medial tangential fields the maximum distance of the heart contour to the posterior field border was measured; this value was scored as the Maximum Heart Distance. The lymph node target volume receiving more than 85% of the prescribed dose was on average 99% for both locoregional irradiation techniques. The breast PTV receiving more than 95% of the prescribed dose was generally smaller using Technique A (mean: 90%, range: 69-99%) than using Technique B (mean: 98%, range: 82-100%) or for the tangential field technique (mean: 98%, range: 91-100%). NTCP values for excess cardiac mortality due to acute myocardial ischemia varied considerably between patients, with minimum and maximum values of 0.1 and 7.5% (Technique A), 0.1 and 5.8% (Technique B) and 0.0 and 6.1% (tangential tech.). The NTCP values were on average significantly higher (P<0.001) by 1.7% (Technique A) and 1.0% (Technique B) when locoregional breast irradiation was given, compared with irradiation of the left breast only. The NTCP values for the tangential field technique could be estimated using the Maximum Heart Distance. NTCP values for radiation pneumonitis were very low for all techniques; between 0.0 and 1.0%. Technique B results in a good coverage of the breast and locoregional lymph nodes, while Technique A sometimes results in an underdosage of part of the target volume. Both techniques result in a higher probability of heart complications compared with tangential irradiation of the breast only. Irradiation toxicity for the lung is low in all techniques. The Maximum Heart Distance is a simple and useful parameter to estimate the NTCP values for cardiac mortality for tangential breast irradiation.


Different effects of raised [K+]o on membrane potential and contraction in mouse fast- and slow-twitch muscle.
Increasing extracellular K+ concentration ([K+]o) from 4 to 7-14 mM reduced both tetanic force and resting membrane potential (Em) in isolated slow-twitch soleus and fast-twitch extensor digitorum longus (EDL) muscles of the mouse. The tetanic force-[K+]o relationships showed a greater force loss over 8-11 mM [K+]o in soleus than EDL, mainly because the Em was 2-3 mV less negative at each [K+]o in soleus. The tetanic force-resting Em relationships show that force was reduced in two phases: phase 1 (Em < -60 mV), a 20% force decline in which the relationships superimposed in soleus and EDL, and phase 2 (Em -60 to -55 mV), a marked force decline that was steeper in EDL than soleus. Additionally in phase 2, longer stimulation pulses restored tetanic force; the twitch force-stimulation strength relationship was shifted toward higher voltages; caffeine, a myoplasmic Ca2+ concentration elevator, increased maximum force; and twitch force fell abruptly. We suggest that 1) the K(+)-depressed force is due to reduced Ca2+ release resulting from an altered action potential profile (phase 1) and inexcitable fibers due to an increased action potential threshold (phase 2), and 2) K+ contributes to fatigue in both fast- and slow-twitch muscle when it causes depolarization to about -60 mV.


The interaction between epidural 2-chloroprocaine and morphine: a randomized controlled trial of the effect of drug administration timing on the efficacy of morphine analgesia.
The efficacy and duration of epidural morphine analgesia is diminished when administered after 2-chloroprocaine compared with lidocaine. The mechanism of the interaction between 2-chloroprocaine and morphine is unknown. Possible explanations include differences in the latency and duration of action of the two drugs or opioid receptor antagonism. We hypothesized that administration of epidural morphine 30 min before the initiation of 2-chloroprocaine anesthesia would result in postoperative analgesia of similar duration and quality to that achieved by epidural morphine after the initiation of lidocaine anesthesia in patients undergoing postpartum tubal ligation. Subjects undergoing bilateral postpartum tubal ligation after vaginal delivery with epidural analgesia were randomized to one of three groups. Subjects received epidural morphine or saline 30 min before the initiation of analgesia with 3% 2-chloroprocaine (two groups) or 2% lidocaine (one group), and at the time of surgical incision, they received either epidural saline or morphine. The duration of analgesia was defined as the time from morphine administration until the first request for supplemental analgesia. Duration of epidural morphine analgesia was compared among groups using Kaplan-Meier survival analysis and the log-rank test. Administration of epidural morphine 30 min before the initiation of 2-chloroprocaine anesthesia (n = 29) resulted in a longer median duration of analgesia (28.6 h [95% CI 4.4-52.7]) compared with the administration of morphine after 2-chloroprocaine anesthesia (n = 30) (2.2 h [95% CI 0-4.8]) (P = 0.006). The median duration of analgesia observed when morphine was administered before 2-chloroprocaine was similar to that observed when morphine was administered after initiation of lidocaine anesthesia (n = 28) (25.8 h [95% CI 10.7-40.9]) (P = 0.83). Pain scores were not different in the postanesthesia care unit, but were higher on admission to the postpartum unit in the subjects receiving morphine after 2-chloroprocaine. Supplemental morphine equivalents administered in the first 48 h were similar among groups and there were no differences in opioid-related side effects. This study demonstrates that administration of epidural morphine 30 min before epidural anesthesia with 2-chloroprocaine provides a similar duration of analgesia as epidural morphine after epidural lidocaine anesthesia. This suggests that the observed interaction between epidural morphine and 2-chloroprocaine is a result of differences in latency and duration of action of the two drugs, or that the administration of morphine before 2-chloroprocaine effectively blocks a receptor site antagonism.


Dissection with harmonic scalpel versus cold instruments in parotid surgery.
The harmonic scalpel (HS) has been used successfully in several head and neck surgical procedures. Some authors highlighted its advantages in reducing operative time, blood loss, and damages to surrounding tissue. In our study, we compared the results obtained during parotidectomy using the HS with the traditional approach to determine the benefits of the HS. 130 patients with benign parotid tumors were enrolled and randomized into two groups for this prospective study. 63 patients underwent HS parotidectomy, and 67 patients received a parotidectomy using cold instruments and bipolar electrocautery hemostatic control (CI). 20 HS and 2 CI patients did not meet the inclusion criteria requirements, and were excluded. The admission time was significantly shorter in the HS group than the CI group (3.9 ± 1.2 days and 4.7 ± 1.4 days, respectively, p < 0.01). In the early post-operative period, 84% of HS patients and 60% of CI cases showed no facial nerve impairment (p = 0.01). Significantly more CI patients than HS patients showed the onset of Frey's syndrome (29% and 9%, respectively, p = 0.01). Multivariate stepwise regression analysis confirmed the reduction in admission length (Odds Ratio (OR): 0.62; p = 0.02) and the lower risk of Frey's syndrome (OR: 0.29; p = 0.04) in HS compared to CI parotidectomies. In parotid surgery, the HS is useful in preventing Frey's syndrome and reducing early transitory facial nerve dysfunction and admission times, and results in decreased medical costs and increased quality of life.


Asynchronous P300-based brain-computer interfaces: a computational approach with statistical models.
Asynchronous control is an important issue for brain-computer interfaces (BCIs) working in real-life settings, where the machine should determine from brain signals not only the desired command but also when the user wants to input it. In this paper, we propose a novel computational approach for robust asynchronous control using electroencephalogram (EEG) and a P300-based oddball paradigm. In this approach, we first address the mathematical modeling of target P300, nontarget P300, and noncontrol signals, by using Gaussian distribution models in a support vector margin space. Furthermore, we derive a method to compute the likelihood of control state in a time window of EEG. Finally, we devise a recursive algorithm to detect control states in ongoing EEG for online application. We conducted experiments with four subjects to study both the asynchronous BCI's receiver operating characteristics and its performance in actual online tests. The results show that the BCI is able to achieve an averaged information transfer rate of approximately 20 b/min at a low false positive rate (one event per minute).


Angiogenesis and gene therapy in man: dream or reality?
Preclinical studies indicate that angiogenic growth factors can stimulate the development of collateral arteries in animal models of peripheral or myocardial ischaemia, a concept termed 'therapeutic angiogenesis'. The goal of this review is to provide a brief overview of the advantages and disadvantages of gene versus recombinant protein therapy for therapeutic angiogenesis. We also discuss different options for delivering genes to patients, including plasmids and modified viral vectors. Recently, the safety and potential utility of gene therapy for ischaemic disease were demonstrated in 3 clinical trials involving the delivery of plasmid DNA encoding the 165 amino acid isoform of human vascular endothelial growth factor (phVEGF165), a factor that specifically promotes the proliferation and migration of vascular endothelial cells. Two trials involved the administration of phVEGF165 for peripheral arterial disease. In one trial, the plasmid was administered to the arterial wall from a hydrogel-coated angioplasty balloon, while a second trial examined the direct injection of phVEGF165 into the skeletal muscle of the affected limb. More recently, phVEGF165 was directly injected into ischaemic myocardium. In all these trials, it appears that administration of phVEGF165 led to improvements in tissue perfusion.


Early and late losses of motor units after poliomyelitis.
Motor unit number estimation was employed to assess muscle innervation in 76 patients with prior poliomyelitis. Of the 68 patients who were < 70 years of age, new musculoskeletal symptoms had appeared in all but four; the mean latent interval was 38.0 +/- 10.1 years. As expected, there was a high incidence of muscles exhibiting denervation in previously affected limbs (87%). However, the incidence in supposedly unaffected limbs was also high (65%). Significant differences in the degree of denervation were found between muscles of the same hands and feet. Judged on the basis of their potential amplitudes, the surviving motor units in partially denervated muscles tended to be enlarged. The enlargement was proportional to the extent of the denervation and was comparable to that found in amyotrophic lateral sclerosis. In some muscles, possibly those innervated by failing motor neurons, motor-unit enlargement was not present. Needle examination confirmed the high incidences of denervation in affected and allegedly unaffected limbs. Of the 188 muscles with EMG features of chronic denervation, only nine exhibited fibrillations or positive sharp waves (4.8%). Ninety-five muscles of 18 patients were studied a second time after an interval of 2 years. Overall, there was a 13.4% reduction in motor-unit number and a 18.4% diminution in M-wave amplitude (P < 0.001). The rate of motor-unit loss was twice that occurring in healthy subjects aged > 60 years. Analysis of individual patients indicated that some were deteriorating more rapidly than others. These studies confirm that denervation progresses in patients with prior poliomyelitis in both clinically affected and unaffected muscles, and indicate that this progression is more rapid than that occurring in normal ageing.


Dry eyes: etiology and management.
Until recently, the cause of dry eye syndrome was uncertain and the treatment was palliative. Since discovering that dry eyes are caused by inflammation, there has been an abundance of research focusing on anti-inflammatory therapies, other contributing causes, and better diagnostic testing. This review summarizes some of the interesting published research on ocular surface disease over the past year. The definition of dry eye now highlights the omnipresent symptom of blurry vision. The re-evaluation of ocular surface staining, tear meniscus height, and visual change will allow for a better diagnosis and understanding of dry eyes. Punctal plugs, and oral and topical anti-inflammatory use will strengthen our arsenal against ocular surface disease. Major progress has occurred in the past few years in gaining a better understanding of the etiology of dry eye syndrome, which will inevitably lead to more effective therapeutic options.


Rosette-forming glioneuronal tumor: pathology case report.
Rosette-forming glioneuronal tumor is a newly described mixed glial and neuronal tumor. We describe two cases and review the literature to better characterize this entity. Patients were surgically treated, and tumors were diagnosed by light microscopy and immunohistochemistry using the avidin-biotin complex method. PubMed was searched for previously reported cases. Patient 1 was a 38-year-old woman who presented with headaches and no neurological abnormality. Magnetic resonance imaging showed a solid mass in the fourth ventricle. Subtotal excision of the mass caused transient gait ataxia. Patient 2 was a 51-year-old woman with dizziness who fell and sustained head trauma. Magnetic resonance imaging revealed a right paramedian cerebellar cystic and nodular mass and a separate nodule in the vermis, which were excised gross totally with no morbidity. Microscopic examination showed neuroepithelial tumors composed of neurocytic cells focally forming well-defined rosettes that were immunopositive for neuronal markers and of elongated, glial fibrillary acidic protein-immunoreactive astrocytes. No histological anaplasia was present. Both patients were well 18 and 8 months after surgery, respectively. Eighteen rosette-forming glioneuronal tumors were identified with the literature search. These are tumors of young adulthood (range, 12-59 yr) usually in or close to the fourth ventricle. Histologically, they are low-grade, although multiple foci or local extension may prevent total excision and account for some recurrences. On imaging, they are cystic, solid, or both, with minimal perilesional edema or mass effect. They are composed of neurocytic and glial elements, probably arising from a common progenitor in the subependymal plate, and need to be differentiated from a variety of glioneuronal tumors.


Distribution of picophytoplankton communities from brackish to hypersaline waters in a South Australian coastal lagoon.
Picophytoplankton (i.e. cyanobacteria and pico-eukaryotes) are abundant and ecologically critical components of the autotrophic communities in the pelagic realm. These micro-organisms colonized a variety of extreme environments including high salinity waters. However, the distribution of these organisms along strong salinity gradient has barely been investigated. The abundance and community structure of cyanobacteria and pico-eukaryotes were investigated along a natural continuous salinity gradient (1.8% to 15.5%) using flow cytometry. Highest picophytoplankton abundances were recorded under salinity conditions ranging between 8.0% and 11.0% (1.3 x 106 to 1.4 x 106 cells ml-1). Two populations of picocyanobacteria (likely Synechococcus and Prochlorococcus) and 5 distinct populations of pico-eukaryotes were identified along the salinity gradient. The picophytoplankton cytometric-richness decreased with salinity and the most cytometrically diversified community (4 to 7 populations) was observed in the brackish-marine part of the lagoon (i.e. salinity below 3.5%). One population of pico-eukaryote dominated the community throughout the salinity gradient and was responsible for the bloom observed between 8.0% and 11.0%. Finally only this halotolerant population and Prochlorococcus-like picocyanobacteria were identified in hypersaline waters (i.e. above 14.0%). Salinity was identified as the main factor structuring the distribution of picophytoplankton along the lagoon. However, nutritive conditions, viral lysis and microzooplankton grazing are also suggested as potentially important players in controlling the abundance and diversity of picophytoplankton along the lagoon. The complex patterns described here represent the first observation of picophytoplankton dynamics along a continuous gradient where salinity increases from 1.8% to 15.5%. This result provides new insight into the distribution of pico-autotrophic organisms along strong salinity gradients and allows for a better understanding of the overall pelagic functioning in saline systems which is critical for the management of these precious and climatically-stress ecosystems.


Effects of P-Aminophenyl Dichloroarsine on Reduced High-affinity [3H]Nicotine Binding Sites from Chick Brain: A Covalent, Yet Reversible, Agent for Neuronal Nicotinic Receptors.
Neuronal nicotinic acetylcholine receptor (nAChR) alpha-subunits contain a conserved disulphide that is essential for function. Here, we have examined the effects of sulphydryl redox reagents on [3H]nicotine binding to chick brain nAChR immunoisolated with the monoclonal antibody mAb35. The disulphide reducing agent, dithiothreitol (DTT), inhibited [3H]nicotine binding [50% inhibitory concentration (IC50)=146 microM] but this effect was reversed (93 +/- 1.5%) by subsequent reoxidation with 1 mM dithio-bis(nitrobenzoic acid) (DTNB). The trivalent arsenical, p-aminophenyl dichloroarsine (APA), which reacts with pairs of spatially close sulphydryls, was a potent inhibitor of reoxidation by DTNB (IC50=35 nM). However, application of the 'anti-arsenical', 2,3-dimercaptopropane sulphonic acid (DMPS), restored agonist binding after APA treatment (50% effective concentration=120 microM). Paradoxically, DMPS was also found to be a potent oxidizing agent of these receptors. Affinity alkylation of reduced nAChRs with bromoacetylcholine (BAC; 100 microM) irreversibly blocked nicotine binding (>90%). We propose (but have not proven) that APA interacts with the cysteines homologous to Cys192 - 193 in Torpedo AChRs, since APA pretreatment of reduced neuronal receptors protected against irreversible BAC alkylation, as shown by subsequent reversal of DMPS (2 mM; 20 min). This study illustrates the potent and reversible nature of the arsenical's covalent interaction with an isolated nAChR and suggests that modified arsenicals could be useful nAChR probes.


Hematotoxicity testing by cell clonogenic assay in drug development and preclinical trials.
In vitro clonogenic assays have been developed and widely used since many years to investigate the proliferation and the differentiation both of pluripotent haemopoietic stem cells (PHSC) and of the different progenitors of blood cell lineages: megakaryocytes (Colony Forming Unit-Mk) granulocyte -macrophage (CFU-GM), erythrocytes (BFU-E/CFU-E). As these techniques have been introduced, they appeared to be very useful to investigate the pathogenic mechanisms of drug induced blood disorders and also for screening compound during preclinical safety study. Because the integrity of the hematopoiesis is also essential to guarantee the immunological function, the application to the toxicology of these clonogenic assays, provides an essential tool for better understanding the in vivo observation (experimental and clinical) by also helping in predicting the degree of a possible in vivo hematotoxic in drug treated patients. The review introduces to basic concepts on hematopoiesis and on classical evaluation of a drug hematotoxic phenomenon. It describes the application of each clonogenic test to assess the specific hematotoxicity action. Moreover it report the most recent studies on standardisation, prevalidation and validation of such assays and critically reviews a model for predicting myelotoxicity by discussing the main aspects referred to the evaluation of the in vivo acute neutropenia by using the in vitro CFU-GM assay.


Heavy snoring is a risk factor for case fatality and poor short-term prognosis after a first acute myocardial infarction.
Sleep disordered breathing has been associated with an increased risk for developing coronary heart disease. Data on the effects of sleep disordered breathing on case fatality and prognosis of a myocardial infarction are sparse. The present study aimed to investigate a possible relationship of snoring and case fatality and mortality after an acute myocardial infarction. DESIGN, SETTINGS, PATIENTS, AND MEASUREMENTS: In this study, we enrolled 1660 first acute myocardial infarction cases and examined the effects of self- or relative-reported heavy snoring on case fatality and prognosis. The average follow-up time was 8 years, SD = 262 days. There was a variation in the association between snoring and mortality with time, with a strong association in the first 28 days after infarction but not later during the follow-up. Occasional and regular heavy snorers, when compared to those never having heavy snoring, had a 2.04 (95% confidence interval, 1.50 to 2.79) and 3.30 (95% confidence interval, 2.37 to 4.58) hazard ratio for mortality within the first 28 days after controlling for age, gender, obesity, history of diabetes and hypertension, physical activity, smoking, and education, respectively. There was no association between snoring and new myocardial infarction, stroke, or hospitalization for heart failure during the follow-up. Heavy snoring is associated with case fatality and short-term mortality in patients with a first acute myocardial infarction.


Interleukin-6 is a needed proinflammatory cytokine in the prolonged neural activity and transcriptional activation of corticotropin-releasing factor during endotoxemia.
Interleukin-6 (IL-6) is a proinflammatory cytokine that plays multiple roles in the central nervous system during infections and injuries. Although this molecule is capable of stimulating the release of ACTH and glucocorticoids, it has been demonstrated that a single injection of IL-6 fails to activate the paraventricular nucleus (PVN) neurons that control the hypothalamic-pituitary-adrenal axis. The observation that IL-6 receptor (IL-6R) is up-regulated in the brain during endotoxemia led us to hypothesize that prior induction of IL-6R synthesis could amplify the effect of circulating IL-6 on the neuroendocrine response. Rats received a first iv injection of either bacterial lipopolysaccharide (LPS; 5 microg) or vehicle solution. After a 6-h waiting period, they received a second iv injection of either recombinant rat IL-6 or vehicle solution and were killed 1 h thereafter. Using in situ hybridization, we observed that IL-6R was barely expressed in the PVN under basal conditions, but was rapidly produced in response to LPS. IL-6 itself was also able to induce the synthesis of its own receptor along cerebral blood vessels, and this effect extended to several parenchymal structures, including the PVN, when the cytokine was administrated after LPS. In agreement with our hypothesis, we found that IL-6 injected in LPS-pretreated rats stimulated PVN neurons, as revealed by the expression of CRF primary transcript and c-fos messenger RNA, an immediate early gene used as a marker of cellular activation. A significant increase in plasma corticosterone levels was also found in animals that received iv IL-6 injection after being pretreated 6 h before with the very low dose of LPS. The fact that IL-6 alone or injected after LPS treatment was unable to induce cyclooxygenase-2 synthesis is an argument in favor of a PG-independent mechanism. The relative contribution of IL-6 in stimulating CRF expression in the PVN and neural activity throughout the brain during endotoxemia was also investigated in IL-6-deficient mice after an ip injection of LPS. The endotoxin induced similar c-fos and CRF expression patterns in knockout and wild-type mice, but the expression levels were generally higher and/or lasted longer in wild-type animals. Taken together, physiological changes that may include the induction of IL-6R synthesis seem to be necessary for IL-6 to activate PVN neurons. Moreover, although IL-6 does not appear essential during the early phases of endotoxemia, this cytokine is required during the later phases to prolong the activation of neural cells throughout the brain and to maintain CRF expression in the PVN neurons that control the hypothalamic-pituitary-adrenal axis.


SV40 T antigen interactions with ssDNA and replication protein A: a regulatory role of T antigen monomers in lagging strand DNA replication.
DNA replication is a central process in all living organisms. Polyomavirus DNA replication serves as a model system for eukaryotic DNA replication and has considerably contributed to our understanding of basic replication mechanisms. However, the details of the involved processes are still unclear, in particular regarding lagging strand synthesis. To delineate the complex mechanism of coordination of various cellular proteins binding simultaneously or consecutively to DNA to initiate replication, we investigated single-stranded DNA (ssDNA) interactions by the SV40 large T antigen (Tag). Using single molecule imaging by atomic force microscopy (AFM) combined with biochemical and spectroscopic analyses we reveal independent activity of monomeric and oligomeric Tag in high affinity binding to ssDNA. Depending on ssDNA length, we obtain dissociation constants for Tag-ssDNA interactions (KD values of 10-30 nM) that are in the same order of magnitude as ssDNA binding by human replication protein A (RPA). Furthermore, we observe the formation of RPA-Tag-ssDNA complexes containing hexameric as well as monomeric Tag forms. Importantly, our data clearly show stimulation of primase function in lagging strand Okazaki fragment synthesis by monomeric Tag whereas hexameric Tag inhibits the reaction, redefining DNA replication initiation on the lagging strand.


Effects of Astaxanthin from Litopenaeus Vannamei on Carrageenan-Induced Edema and Pain Behavior in Mice.
Carrageenan produces both inflammation and pain when injected in mouse paws via enhancement of reactive oxygen species formation. We have investigated an effect of astaxanthin extracted from Litopenaeus vannamei in carrageenan-induced mice paw edema and pain. The current study demonstrates interesting effects from astaxanthin treatment in mice: an inhibition of paw edema induced in hind paw, an increase in mechanical paw withdrawal threshold and thermal paw withdrawal latency, and a reduction in the amount of myeloperoxidase enzyme and lipid peroxidation products in the paw. Furthermore the effect was comparable to indomethacin, a standard treatment for inflammation symptoms. Due to adverse effects of indomethacin on cardiovascular and gastrointestinal systems, our study suggests promising prospect of astaxanthin extract as an anti-inflammatory alternative against carrageenan-induced paw edema and pain behavior.


Scanning training in neurological vision loss: case studies.
The purpose of this paper is to describe the response of a Department of Veterans Affairs medical center's development of a rehabilitation program for patients with hemianopsia. Hemianopsia affects significant numbers of troops returning from Afghanistan and Iraq and their neurological vision loss presented unique challenges in developing an appropriate and effective rehabilitation program. A literature review indicated that existing therapies lacked supporting scientific evidence and that traumatic brain injury (TBI)-related vision loss affects large numbers of civilians. The increasing number of patients with TBI-related vision loss necessitated the development of an innovative program which combined elements of therapies that the literature suggested were most promising. In this paper we briefly review the literature, describe the rehabilitation program developed, and present case studies of two patients who incurred vision loss as a result of a motor vehicle accident and a gunshot wound. The intent of the article is to begin the documentation of our ongoing, evidence-based neurological vision loss rehabilitation program. We also encourage others who do not currently do so to assess the need for implementing vision rehabilitation programs for patients with TBI-related vision loss.


Reproducibility of anterior chamber angle analyses with the swept-source optical coherence tomography in young, healthy Caucasians.
To assess the reproducibility of iridocorneal angle (ICA) analysis in young, healthy Caucasian subjects using swept-source optical coherence tomography (SS-OCT) by determining variability and interobserver agreement between expert and nonexpert observers. Thirty-one healthy volunteers (nonexperts) acquired three consecutive SS-OCT images of the right eyes of their peer nonexperts. Images were analyzed by 31 nonexperts and additionally by three experts, whereby the angle opening distance (AOD) and the trabecular iris space area (TISA) at 500 and 750 μm were calculated. A random intercept model was used to determine the amount of variation between observers. In addition, the intra-observer variability between nonexperts and experts was calculated by determining the coefficient of variation (CV). A significant difference was found in the expert analysis for the nasal and temporal angle in the AOD500 (P = 0.002), AOD750 (P < 0.01), and TISA750 (P < 0.01), and the values AOD500 (P = 0.025), AOD750 (P = 0.012), and TISA500 (P = 0.010) were significantly larger if nonexperts analyzed SS-OCT images. The CV was only significant larger for nonexperts for AOD500 (11.1% vs. 8.7%, P < 0.01). This study demonstrated high reproducibility of angle analysis in young, healthy Caucasian subjects using SS-OCT. Nevertheless, nonexperts obtained significant larger values compared with experts, implying that training is a necessary requirement before analyzing SS-OCT images in ophthalmic practice.


The influenza resistance murine Mx1 gene is constitutively expressed in the epithelia of the gastrointestinal, respiratory and uterine tracts.
The murine Mx1 gene confers specific resistance against influenza in the inbred A2G mice and in vitro have been shown to be inducible with type I but not type II interferons. Contrary to expectation, we found by in situ hybridisation widespread Mx1 expression along the epithelia of the gastrointestinal, uterine and respiratory tracts in uninduced A2G mice. Several lines of evidence, including further enhancement of Mx1 expression during organ culture and gnotobiotic mice analyses, indicated that this apparent constitutive epithelial Mx1 expression was a locally induced response to stimuli present in the respective lumina. This phenomenon may be a feature found in other interferon-inducible and interferon genes.


Molecular-level characterization of elastin-like constructs and human aortic elastin.
This study aimed to characterize the structures of two elastin-like constructs, one composed of a cross-linked elastin-like polypeptide and the other one of cross-linked tropoelastin, and native aortic elastin. The structures of the insoluble materials and human aortic elastin were investigated using scanning electron microscopy. Additionally, all samples were digested with enzymes of different specificities, and the resultant peptide mixtures were characterized by ESI mass spectrometry and MALDI mass spectrometry. The MS(2) data was used to sequence linear peptides, and cross-linked species were analyzed with the recently developed software PolyLinX. This enabled the identification of two intramolecularly cross-linked peptides containing allysine aldols in the two constructs. The presence of the tetrafunctional cross-link desmosine was shown for all analyzed materials and its quantification revealed that the cross-linking degree of the two in vitro cross-linked materials was significantly lower than that of native elastin. Molecular dynamics simulations were performed, based on molecular species identified in the samples, to follow the formation of elastin cross-links. The results provide evidence for the significance of the GVGTP hinge region of domain 23 for the formation of elastin cross-links. Overall, this work provides important insight into structural similarities and differences between elastin-like constructs and native elastin. Furthermore, it represents a step toward the elucidation of the complex cross-linking pattern of mature elastin.


Curcumin blocks Kv11.1 (erg) potassium current and slows proliferation in the infant acute monocytic leukemia cell line THP-1.
Acute Myeloid Leukemia (AML) accounts for approximately one fifth of all childhood leukemia yet is responsible for a significant proportion of morbidity and mortality in this population. For this reason, research to identify novel targets for the development of effective AML therapeutics has intensified in the recent past. The THP-1 cell line, which was originally established from an infant diagnosed with AML, provides an experimental model for functional, pre-clinical therapeutics and target identification studies of AML. Here we show the expression of the voltage gated potassium channel Kv11.1 in THP-1 cells as opposed to normal hematopoietic stem cells. In addition, curcumin, a natural polyphenol derived from the plant Curcuma longa, effectively blocked Kv11.1 activity and also inhibited the proliferation of these cells. Curcumin was rapidly internalized by THP-1 cells and possibly exerts potential growth inhibitory activity by interacting with intracellular epitopes of the ion channel. Inhibition of ionic currents carried by Kv11.1 resulted in depolarization of cell membrane potential. We propose that the inhibition of Kv11.1 activity by curcumin may lead to interference with leukemic cell physiology and consequently the suppression of survival and proliferation of AML cells.


Clinical-pathological characteristics and prognosis of a cohort of oesophageal cancer patients: a competing risks survival analysis.
To determine the clinical course, follow-up strategies, and survival of oesophageal cancer patients using a competing risks survival analysis. We conducted a retrospective and prospective follow-up study. The study included 180 patients with a pathological diagnosis of oesophageal cancer in A Coruña, Spain, between 2003 and 2008. The Kaplan-Meier methodology and competing risks survival analysis were used to calculate the specific survival rate. The study was approved by the Ethics Review Board (code 2011/372, CEIC Galicia). The specific survival rate at the first, third, and fifth years was 40.2%, 18.1%, and 12.4%, respectively. Using the Kaplan-Meier methodology, the survival rate was slightly higher after the third year of follow-up. In the multivariate analysis, poor prognosis factors were female sex (hazard ratio [HR] 1.94; 95% confidence interval [CI], 1.24-3.03), Charlson's comorbidity index (HR 1.17; 95% CI, 1.02-1.33), and stage IV tumours (HR 1.70; 95% CI, 1.11-2.59). The probability of dying decreased with surgical and oncological treatment (chemotherapy and/or radiotherapy) (HR 0.23; 95% CI, 0.12-0.45). The number of hospital consultations per year during the follow-up period, from diagnosis to the appearance of a new event (local recurrences, newly appeared metastasis, and newly appeared neoplasias) did not affect the probability of survival (HR 1.03; 95% CI, 0.92-1.15). The Kaplan-Meier methodology overestimates the survival rate in comparison to competing risks analysis. The variables associated with a poor prognosis are female sex, Charlson's comorbidity score and extensive tumour invasion. Type of follow-up strategy employed after diagnosis does not affect the prognosis of the disease.


Elective cesarean delivery for women with a previous anal sphincter rupture.
The purpose of this study was to evaluate elective cesarean delivery for women with a history of anal sphincter rupture. The effectiveness of cesarean delivery in parous women with a previous anal sphincter rupture was determined by decision analysis. The outcomes were excess cesarean deliveries and morbidity and mortality rates per prevented case of anal incontinence. We needed 2.3 cesarean deliveries to prevent one case of anal incontinence. A woman who chooses a cesarean delivery has a 11.3% risk of morbidity compared with a 4.2% risk for vaginal delivery (relative risk, 2.7; 95% CI, 2.6-2.8; P<.001). The relative risk for maternal death from a cesarean delivery is 2.6 (95% CI, 1.5-4.5; P<.001). Continent women with a previous anal sphincter rupture who are delivered vaginally are at high risk for permanent anal incontinence. Cesarean delivery will prevent most cases of anal incontinence, although marginally increasing maternal risk. The increased risk may be justified by the potential benefits. Patients should be counseled on these risks and benefits.


New lessons learned in T-PLL: results from a prospective phase-II trial with fludarabine-mitoxantrone-cyclophosphamide-alemtuzumab induction followed by alemtuzumab maintenance.
Clinical trials in T-cell prolymphocytic leukemia (T-PLL) are scarce. Based on a precursor study testing fludarabine, mitoxantrone, and cyclophosphamide followed by alemtuzumab (FMC-A), we aimed to improve this regimen by upfront combining subcutaneous (s.c.) alemtuzumab with FMC for four cycles followed by an alemtuzumab-maintenance (FMCA + A). This prospective multicenter phase-II trial assessed response, survival, and toxicity of that regimen administered to pretreated (n = 4) and treatment-naïve (n = 12) T-PLL patients. The best overall response rate after FMCA was 68.8% (n = 11) including five CRs (31.3%) and six PRs (37.5%). Six patients entered the alemtuzumab-maintenance. Median overall and progression-free survival was 16.7 and 11.2 months, respectively. Hematologic toxicities were the most frequent grade 3/4 side effects. A reduced incidence of CMV-reactivations was attributed to the prophylactic administration of valganciclovir. Overall, FMCA + A did not improve the efficacy of the FMC-A-regimen or of single i.v. alemtuzumab. It suggests that a chemotherapy backbone prevents efficient alemtuzumab dosing and confirms that intravenous alemtuzumab is to be preferred over its s.c. route in T-PLL. ClinicalTrials.gov identifier: NCT01186640.


[Strategy of treatment for bleeding of portal genesis].
Based on a comparative analysis of results of the surgical treatment of bleedings from varicose veins of the liver and stomach, the authors recommend differential decision for the variants of surgical interventions at different periods from the moment of the appearance of bleeding and depending on the severity of impairments of the liver functions.


Long-term results after lateral canthal advancement for unilateral coronal synostosis.
The procedure of lateral canthal advancement was carried out to correct unilateral coronal synostosis in 39 consecutive patients at The Hospital for Sick Children during a 20-year period. Thirty-seven children were available for follow-up study, and results were considered to be good or excellent in 81% of them. Liberation of the supraorbital margin and release of stenotic skull base sutures are crucial for a successful result. The severity of the process and the age of the patient were not restrictive factors in regard to indication for this procedure. Some patients showed a continuous improvement with time. Morbidity was minimal. It is suggested that lateral canthal advancement is the procedure of choice for unilateral coronal synostosis.


Fast and Slow-Growing Management Systems: Characterisation of Broiler Caecal Microbiota Development throughout the Growing Period.
Caecal microbiota and its modulation play an important role in poultry health, productivity and disease control. Moreover, due to the emergence of antimicrobial-resistant bacteria, society is pressing for a reduction in antibiotic administration by finding effective alternatives at farm level, such as less intensified production systems. Hence, the aim of this study was to characterise the caecal microbiota in two different broiler management systems, fast and slow-growing, using 16S rRNA sequencing analysis. To this end 576 broilers were reared in two different management systems (fast and slow-growing). Results showed that Firmicutes represented the dominant phylum for both systems. At the onset, Proteobacteria was the second prevalent phylum for fast and slow-growing breeds, outnumbering the Bacteroidetes. However, during the rest of the production cycle, Bacteroidetes was more abundant than Proteobacteria in both groups. Finally, regardless of the management system, the most predominant genera identified were Oscillospira spp., Ruminococcus spp., Coprococcus spp., Lactobacillus spp. and Bacteroides spp. In conclusion, fast and slow-growing broiler microbiota are in constant development throughout rearing, being relatively stable at 21 days of age. Regarding the genus, it should be noted that the three most abundant groups for both systems, Ruminococcus spp., Lactobacillus spp. and Bacteroides spp., are related to better productive performance and intestinal health.


Does a detectable HCV RNA at the end of DAA therapy herald treatment failure?
The study aimed to assess the phenomenon of achieving sustained virologic response (SVR) in patients with detectable ribonucleic acid (RNA) of hepatitis C virus (HCV) at the end of treatment (ET) with direct-acting antivirals (DAA), find how this is affected by the type of regimen, and how patients experiencing this differed from non-responders with detectable HCV RNA at the ET. The study included all consecutive patients with detectable HCV RNA at the ET selected from the EpiTer-2 database, a retrospective national multicentre project evaluating antiviral treatment in HCV-infected patients in 2015-2023. Of the 16106 patients treated with IFN-free regimens with available HCV RNA assessment at the ET and at follow-up 12 weeks after treatment completion (FU), 1253 (7.8%) had detectable HCV RNA at the ET, and 1120 of them (89%) finally achieved SVR. This phenomenon was significantly more frequent in pangenotypic regimens, 10.3% vs. 4.7% in genotype-specific options (p < 0.001), and the highest proportion was documented for glecaprevir/pibrentasvir (13.7%), and velpatasvir/sofosbuvir ± ribavirin (6.9%). Patients ET + FU- treated with these two pangenotypic regimens (n = 668) had less advanced liver disease, were less frequently infected with genotype (GT) 3, and were significantly more likely to be treatment-naïve than 61 non-responders. We documented 7.8% rate of patients with detectable HCV RNA at the ET, of whom 89% subsequently achieved SVR, significantly more frequently in the population treated with pangenotypic regimens. Less severe liver disease, more often GT3 infection, and a higher percentage of treatment-naive patients distinguished this group from non-responders.


Pregnancy associated TMA in 13-year-old patient successfully treated with Eculizumab: case report.
This report introduces an unusual cause of kidney failure in a previously healthy pediatric patient. She developed thrombotic microangiopathy (TMA) that was diagnosed post-partum, requiring dialysis and eculizumab, with eventual recovery of kidney function ([chronic kidney disease (CKD) stage 3]. The patient was induced at term due to preeclampsia, with delivery complicated by severe postpartum hemorrhage from uterine atony. She continued to have severe hypertension post-delivery and further developed acute kidney injury (AKI) with decreased urinary output and respiratory distress requiring dialysis therapy. Labs revealed hemolysis with elevated lactate dehydrogenase, low haptoglobin, anemia, and thrombocytopenia, but otherwise unremarkable immunology labs. Once clinically stabilized the patient underwent kidney biopsy, which was consistent with TMA. Treatment was initiated with eculizumab, a monoclonal antibody for terminal complement blockade. Her clinical status improved (including markers of hemolysis and inflammation) with kidney replacement therapy and complement blockade. On discharge, she had increasing urine output and was prescribed 3 day per week hemodialysis and twice monthly eculizumab infusions. By 6 weeks post-delivery, hemodialysis was discontinued and her eculizumab was weaned to monthly infusions. Eculizumab was discontinued at 12 months postpartum. Genetic testing for mutations of the complement system was negative. The patient has residual stage 3 CKD with stable kidney function, requiring two agents for blood pressure control, including an ACE inhibitor for antiproteinuric effect. This case report showcases an unusual cause of renal failure in a pediatric patient due to TMA in the post-partum period. She required intermittent hemodialysis (iHD) for a brief period, however she was treated successfully with eculizumab that was able to be weaned off 1 year after delivery. She has residual stage 3 CKD and no further signs or symptoms of TMA.


Characterizing multiple independent behavioral correlates of cell firing in freely moving animals.
The heterogeneous sampling of behavioral states by freely moving animals hinders our ability to relate neuronal firing rates to behavioral variables by introducing dependencies between them. We specifically consider the animal's location and orientation, although our analyses may generalize to other behavioral variables, such as speed of movement. A maximum-likelihood approach is presented for producing estimates of the separate histograms relating firing rate to multiple independent causes. Examples show that the method can be used to avoid the artifactual behavioral correlates of place and head direction-cell firing produced by standard analyses; to characterize the independent influences of both location and orientation in a third cell type (Cacucci et al., 2004); and to demonstrate the location-independence of the directional firing of head-direction cells.


Engagement of the EP2 prostanoid receptor closes the K+ channel KCa3.1 in human lung mast cells and attenuates their migration.
Human lung mast cells (HLMC) express the Ca(2+)-activated K(+) channel K(Ca)3.1, which plays a crucial role in their migration to a variety of diverse chemotactic stimuli. K(Ca)3.1 activation is attenuated by the beta(2)-adrenoceptor and the adenosine A(2A) receptor through a G(s)-coupled mechanism independent of cyclic AMP. Prostaglandin E(2) promotes degranulation and migration of mouse bone marrow-derived mast cells through the G(i)-coupled EP(3) prostanoid receptor, and induces LTC(4) and cytokine secretion from human cord blood-derived mast cells. However, PGE(2) binding to the G(s)-coupled EP(2) receptor on HLMC inhibits their degranulation. We show that EP(2) receptor engagement closes K(Ca)3.1 in HLMC. The EP(2) receptor-specific agonist butaprost was more potent than PGE(2) in this respect, and the effects of both agonists were reversed by the EP(2) receptor antagonist AH6809. Butaprost markedly inhibited HLMC migration induced by chemokine-rich airway smooth muscle-conditioned media. Interestingly, PGE(2) alone was chemotactic for HLMC at high concentrations (1 microM), but was a more potent chemoattractant for HLMC following EP(2) receptor blockade. Therefore, the G(s)-coupled EP(2) receptor closes K(Ca)3.1 in HLMC and attenuates both chemokine- and PGE(2)-dependent HLMC migration. EP(2) receptor agonists with K(Ca)3.1 modulating function may be useful for the treatment of mast cell-mediated disease.


Ellagic acid exerts anti-proliferation effects via modulation of Tgf-β/Smad3 signaling in MCF-7 breast cancer cells.
Ellagic acid has been shown to inhibit tumor cell growth. However, the underlying molecular mechanisms remain elusive. In this study, our aim was to investigate whether ellagic acid inhibits the proliferation of MCF- 7 human breast cancer cells via regulation of the TGF-β/Smad3 signaling pathway. MCF-7 breast cancer cells were transfected with pEGFP-C3 or pEGFP-C3/Smad3 plasmids, and treated with ellagic acid alone or in combination with SIS3, a specific inhibitor of Smad3 phosphorylation. Cell proliferation was assessed by MTT assay and the cell cycle was detected by flow cytometry. Moreover, gene expression was detected by RT-PCR, real-time PCR and Western blot analysis. The MTT assay showed that SIS3 attenuated the inhibitory activity of ellagic acid on the proliferation of MCF-7 cells. Flow cytometry revealed that ellagic acid induced G0/G1 cell cycle arrest which was mitigated by SIS3. Moreover, SIS3 reversed the effects of ellagic acid on the expression of downstream targets of the TGF-β/Smad3 pathway. In conclusion, ellagic acid leads to decreased phosphorylation of RB proteins mainly through modulation of the TGF-β/Smad3 pathway, and thereby inhibits the proliferation of MCF-7 breast cancer cells.


Deep brain stimulation in area LC controllably triggers auditory phantom percepts.
Tinnitus is predominantly viewed as the consequence of dysfunctional hyperactivity, plastic change, or synchronized oscillations in the central auditory system. An alternative to the current auditory-centric view of auditory phantom perception is the basal ganglia-centric view. Recent electrical stimulation experiments in area LC, a locus of the caudate nucleus positioned at its anterior body, has shown loudness modulation of existing tinnitus percepts. To demonstrate that auditory phantoms are gated by the dorsal striatum. Electrical stimulation in area LC via a deep brain stimulation lead was performed in 6 interactive adult subjects (3 with and 3 without chronic tinnitus) undergoing surgery to treat movement disorders. Tinnitus loudness was rated on a 0 to 10 scale, sound quality was described, and localization was referenced to 1 or both ears. Short-term area LC stimulation triggered new phantom tones, clicks, and frequency modulated sounds in 5 subjects and altered sound quality of an existing tinnitus percept in 1 subject. The results of this study indicate that perceptual awareness of auditory phantoms is contingent on satisfying a permission condition controlled by the dorsal striatum. Potential auditory phantoms are not automatically gated to reach perceptual awareness. A phantom percept gate control model is proposed. Neuromodulation of area LC can trigger temporary gate dysfunction and reversibly release new phantoms for conscious awareness. Restoration of restrictive dorsal striatal gate function to treat problematic phantom percepts may be realized by adopting long-term area LC neuromodulation and choosing optimal stimulation parameters.


Novel mutations introduced at the beta-site of amyloid beta protein precursor enhance the production of amyloid beta peptide by BACE1 in vitro and in cells.
Abnormal production and accumulation of amyloid-beta peptide (Abeta) plays a major role in the pathogenesis of Alzheimer's disease (AD). beta-secretase (BACE1) is responsible for the cleavage at thebeta-site in amyloid beta protein precursor (AbetaPP/APP) to generate the N-terminus of Abeta. Here we report the stepwise identification and characterization of a novel APP-beta-site mutant, "NFEV" (APP_NFEV) in vitro and in cells. In vitro, the APP_NFEV exhibits 100-fold enhanced cleavage rate relative to the "wild-type" substrate (APPwt) and 10-fold increase relative to the Swedish-type mutation variant (APPsw). In cells, it was preferably cleaved among 24 APP beta-site mutations tested. More importantly, the APP_NFEV mutant failed to generate any detectable Abeta peptides in BACE1-KO mouse fibroblast cells. The production of Abeta peptides was restored by co-transfecting human BACE1, demonstrating that BACE1 is the only enzyme responsible for the processing of APP_NFEV in these cells. Analysis of APP_NFEV cleavage products secreted in the media revealed that in cells BACE1 cleaves APP_NFEV at the position between NF and EV, identical to that observed in vitro. A BACE inhibitor blocked the processing of the APP_NFEV beta-site in vitro and in cells. Our data indicates that the "NFEV" mutant is not only an enhanced substrate for BACE1 in vitro, but also a specific substrate for BACE1 in cells.


Comparative physiological responses of Solanum nigrum and Solanum torvum to cadmium stress.
• Under cadmium (Cd) stress, Solanum nigrum accumulated threefold more Cd in its leaves and was tolerant to Cd, whereas its low Cd-accumulating relative, Solanum torvum, suffered reduced growth and marked oxidative damage. However, the physiological mechanisms that are responsible for differential Cd accumulation and tolerance between the two Solanum species are largely unknown. • Here, the involvement of antioxidative capacity and the accumulation of organic and amino acids in response to Cd stress in the two Solanum species were assessed. • Solanum nigrum contains higher antioxidative capacity than does S. torvum under Cd toxicity. Metabolomics analysis indicated that Cd treatment also markedly increased the production of several organic and amino acids in S. nigrum. Pretreatment with proline and histidine increased Cd accumulation; moreover, pretreatment with citric acid increased Cd accumulation in leaves but decreased Cd accumulation in roots, which indicates that its biosynthesis could be linked to Cd long-distance transport and accumulation in leaves. • Our data provide novel metabolite evidence regarding the enhancement of citric acid and amino acid biosynthesis in Cd-treated S. nigrum, support the role of these metabolites in improving Cd tolerance and accumulation, and may help to provide a better understanding of stress adaptation in other Solanum species.


[Implementation of a continuum of care for people living with HIV/AIDS in Hanoi (Vietnam)].
Caring for people living with HIV/AIDS (PLWHA) encompasses various tasks, from prevention to palliative care. It involves a set of consistent and coordinated actions. This article presents the first free-of-charge management programme including antiretroviral treatment in Vietnam (as opposed to research and evaluation programmes). It was launched in 2004 in Hanoi. Our study was conducted in 2003-2004 as part of a collaborative research programme led by IRD (Research Institute for Development) and the National Economic University in Hanoi and was funded by ESTHER (Together for a Therapeutic Solidarity in Hospital Network) group. Data collection included 68 qualitative interviews with patients, members of their families and members of the hospital staff, observations of outpatient consultations, and analysis of inpatient files. The results show that patients, their families and hospital staff members all perceive a comprehensive care and treatment programme as very important and consider that it should include social and psychological care as well as an integrated set of actions involving various types of participants. Outpatient and inpatient care are closely linked: they take place in the same hospital department, they involve patients with similar social and demographic characteristics marked by multiple risk behaviours and recourse to several kinds of healthcare services. The observation of outpatient consultations showed the limitations of strictly biomedical care to which social and psychological care were added only lately. One of the principal difficulties is patients' difficulties in keeping their outpatient appointments. Overall, patients consider themselves lucky to able to receive care and treatment with antiretroviral drugs. They nevertheless complain about the lack of social and psychological support, which they expect should help them to tolerate and adapt to their biomedical treatment and to include counselling and information about this treatment and its consequences. Hospital staff with the greatest contact with PLWHA report more frequent attempts to avoid this contact. This stigmatisation is due to lack of information, failure to implement workplace safety measures, and to pejorative representations of HIV/AIDS. Official and unofficial discourse still follows the Ministry of Health in associating HIV/AIDS with drug use and commercial sex, and HIV/AIDS prevention and control policy is still linked to the "social evils" policy. Hospital staff also emphasized the importance of community care for PLWHA in their interviews. Informal care for PLWHA by family, close relatives, close friends and members of non-official groups complements hospital care, which is sometimes limited to its biomedical component and provides the material, moral, financial, social, economic and relational care essential for PLWHA and their close relatives and friends. This informal care has also some pernicious effects and leads to internal contradictions due to the multiple social roles played by the many and various participants involved. HIV/AIDS prevention and control policy relies on a series of choices between more specificity through vertical programmes specialised in HIV/AIDS and the synergy that can develop through more integrated health services. Vietnam has developed links between HIV/AIDS prevention and control programmes on the one hand, and harm reduction programmes for injecting drug users (access to substitution products such as methadone) and condom distribution, on the other. Nonetheless, HIV/AIDS prevention and control policy faces difficulties in reaching its objectives. The results of this policy, intended to help achieve Millennium Development Goal (MDG) n degrees 6, depends partly on the success for other MDGs, including the fight against poverty, the promotion of gender equality and empowerment of women, and the improvement of reproductive health. To be able to succeed in implementing the continuum of care necessary for treating HIV/AIDS within its institutions, Vietnam can apply the lessons of international experience, adapted to fit local constraints and the social, cultural and political context. The shortcomings encountered in this endeavour shows how difficult it is for this country to implement such a complex set of measures at an accelerated pace. They should not, however, hide or minimize the great efforts, the vigour, and the capacity to adapt already demonstrated by local participants.


Tamoxifen decreases the rate of proliferation of rat vascular smooth-muscle cells in culture by inducing production of transforming growth factor beta.
Tamoxifen selectively and reversibly decreased the rate of proliferation of adult rat aortic vascular smooth-muscle cells (VSMCs). Half-maximal inhibition of proliferation occurred at 2-5 microM tamoxifen for VSMCs and at > 50 microM for adventitial fibroblasts. The cell cycle time for all the VSMCs in the population was increased from 35 +/- 2 h to 54 +/- 4 h in the presence of 33 microM tamoxifen. Tamoxifen did not affect the time of entry into DNA synthesis, but delayed arrival at mitosis by > 24 h. It therefore extended the duration of the G2-to-M phase of the cell cycle. However, the rate of proliferation of VSMCs was not decreased by tamoxifen (at concentrations up to 50 microM) in the presence of neutralizing antibody to transforming growth factor beta (TGF-beta). The level of mRNA for TGF-beta 1 in VSMCs was strongly induced by 10 microM tamoxifen, and TGF-beta activity in conditioned medium from tamoxifen-treated cells was more than 50-fold higher than from control cells. Tamoxifen therefore extended the G2-to-M phase of the cell cycle in VSMCs by increasing TGF-beta activity in the culture.


Full-thickness macular hole and macular telangiectasia in a child with Coats' disease.
A 9-year-old boy presented with a 6-month history of decreased vision in his left eye. The patient's retinal examination was significant for telangiectatic vessels involving the posterior pole and the superior and inferior post-equatorial regions, circinate intraretinal exudation at the macula, and a full-thickness macular hole that was confirmed by optical coherence tomography. Chronic exudation from the anomalous vessels may be involved in the pathogenesis. Full-thickness macular hole formation may occur in conjunction with Coats' disease in children, which has not been reported to date, to the best of the authors' knowledge.


Is it a sarcoidal foreign-body granuloma or a cutaneous sarcoidosis on a permanent eyebrow make-up?
Permanent make-up is a kind of cosmetic tattoo in which the colorants (pigments) are deposited in dermis after piercing the skin by tiny solid needles. It may cause some adverse effects such as local inflammation, infection, and allergic reactions on the skin and even systemic adverse effects such as sarcoidal reactions. Here the case of a 34-year-old woman who has some yellowish hard shiny papules on her eyebrows after having a permanent make-up is described. The histopathological examinations of the papules are diagnosed as sarcoidal foreign-body reactions. All the laboratory investigations were in normal limits except a mild elevation in angiotensin converting enzyme or ACE level. The lesions mostly improved after topical corticosteroid treatment. Sarcoidal foreign-body reaction due to permanent make-up is discussed with this presentation.


Vascular anatomy and hydrostatic pressure profile in the hamster cheek pouch.
The purpose of this study was to describe the vascular architecture and blood supply to the hamster cheek pouch and to measure the intravascular pressure distribution in the entire pouch. Previous anatomic descriptions have focused either on the vasculature of the facial region or on the microcirculation of the cheek pouch tip. Micropressures in cheek pouch capillaries, terminal arterioles, and small venules have not previously been measured. Cheek pouches were prepared for examination under an intravital microscope using both everted and noneverted methods. Microvessel diameters were measured through a video system using a video micrometer, and intravascular hydrostatic pressures were measured with a servo null micropressure system. Blood vessels in the face and pouch were traced after injection of Microfil into the external maxillary artery. The results indicate that the pouch is supplied directly or indirectly by six small arteries in the neck and face. Fifty percent of the total pressure drop across the cheek pouch vasculature occurs in the small cheek pouch arteries, suggesting that these vessels are potentially important in controlling cheek pouch blood flow. The measurements of microvascular pressures and diameters in this study help clarify apparent discrepancies in similar measurements from previous studies.


Enhancement of glucose transport by vascular endothelial growth factor in retinal endothelial cells.
To investigate effects of vascular endothelial growth factor (VEGF) on glucose transport and GLUT1 glucose transporter expression in primary bovine retinal endothelial cell (BREC) cultures. Glucose transport in control and VEGF-treated BREC cultures was determined by measurement of [14C]-3-O-methylglucose (3MG) uptake. GLUT1 protein and mRNA was determined by Western and Northern blot analyses, respectively. Protein kinase C (PKC) activity was measured in control and VEGF-treated cultures, and glucose transport was determined with and without prior PKC depletion and PKC inhibition. Dose-dependent increases in 3MG uptake were seen in the VEGF-treated cultures, with an increase of 69% after a 24-hour exposure to 50 ng/ml VEGF (P < 0.001). Total cellular GLUT1 mRNA or protein, however, was unchanged. Western blot analysis of plasma membrane fractions revealed a 75% increase in plasma membrane GLUT1 in VEGF-treated cultures (P = 0.02), suggesting that the VEGF-stimulated increase in glucose transport was due to a translocation of GLUT1 to the cell membrane. VEGF stimulated a 90% increase in PKC activity in membrane fractions from cultures treated with VEGF, and VEGF-stimulated enhancement of glucose transport was abolished by cellular PKC depletion and by general and PKC beta inhibition. The present study demonstrates VEGF-mediated enhancement of retinal endothelial cell glucose transport and suggests that this increase is due to PKC beta-mediated translocation of cytosolic GLUT1 to the plasma membrane surface. Upregulation of retinal endothelial cell glucose transport by various factors associated with the development of retinopathy may be responsible for the metabolic derangements observed in the diabetic inner blood-retinal barrier in vivo.


Creation and Implementation of a Mastery Learning Curriculum for Emergency Department Thoracotomy.
Emergency department thoracotomy (EDT) is a lifesaving procedure within the scope of practice of emergency physicians. Because EDT is infrequently performed, emergency medicine (EM) residents lack opportunities to develop procedural competency. There is no current mastery learning curriculum for residents to learn EDT. The purpose of this study was to develop and implement a simulation-based mastery learning curriculum to teach and assess EM residents' performance of the EDT. We developed an EDT curriculum using a mastery learning framework. The minimum passing standard (MPS) for a previously developed 22-item checklist was determined using the Mastery Angoff approach. EM residents at a four-year academic EM residency program underwent baseline testing in performing an EDT on a simulation trainer. Performance was scored by two raters using the checklist. Learners then participated in a novel mastery learning EDT curriculum that included an educational video, hands-on instruction, and deliberate practice. After a three-month period, residents then completed initial post testing. Residents who did not meet the minimum passing standard after post testing participated in additional deliberate practice until mastery was obtained. Baseline and post-test scores, and time to completion of the procedure were compared with paired t-tests. Of 56 eligible EM residents, 54 completed baseline testing. Fifty-two residents completed post-testing until mastery was reached. The minimum passing standard was 91.1%, (21/22 items correct on the checklist). No participants met the MPS at the baseline assessment. After completion of the curriculum, all residents subsequently reached the MPS, with deliberate practice sessions not exceeding 40 minutes. Scores from baseline testing to post-testing significantly improved across all postgraduate years from a mean score of 10.2/22 to 21.4/22 (p <0.001). Mean time to complete the procedure improved from baseline testing (6 minutes [min] and 21 seconds [sec], interquartile range [IQR] = 4 min 54 sec - 7 min 51 sec) to post-testing (5 min 19 seconds, interquartile range 4 min 17sec - 6 min 15 sec; p = 0.001). This simulation-based mastery learning curriculum resulted in all residents performing an EDT at a level that met or exceeded the MPS with an overall decrease in time needed to perform the procedure.


Upper gastrointestinal bleeding: etiology and management.
Upper gastrointestinal bleeding is a potentially fatal condition at times due to loss of large volumes of blood. Common sources of upper gastrointestinal bleeding in children include mucosal lesions and variceal hemorrhage (most commonly extra hepatic portal venous obstruction) and, in intensive care settings infections and drugs are other etiological factors associated with bleeding. Massive upper GI bleeding is life threatening and requires immediate resuscitation measures in the form of protection of the airways, oxygen administration, immediate volume replacement with ringer lactate or normal saline, transfusion of whole blood or packed cells and also monitoring the adequacy of volume replacement by central venous lines and urine output. Upper GI endoscopy is an effective initial diagnostic modality to localize the site and cause of bleeding in almost 85-90% of patients. Antacids supplemented by H2- receptor antagonists, proton pump inhibitors and sucralfate are the mainstay in the treatment of bleeding from mucosal lesion. For variceal bleeds, emergency endoscopy is the treatment of choice after initial haemodynamic stabilization of patient. If facilities for endoscopic sclerotherapy (EST) are not available, pharmacotherapy which decreases the portal pressure is almost equally effective and should be resorted to. Shunt surgery is reserved for patients who do not respond to the above therapy. Beta blockers combined with sclerotherapy have been shown to be the most effective therapy in significantly reducing the risk of recurrent rebleeding from varices as well as the death rates, as compared to any other modality of treatment. Based on studies among adult patients, presence of shock, co-morbidities, underlying diagnosis, presence of stigmata of recent hemorrhage on endoscopy and rebleeding are independent risk factors for mortality due to upper GI bleeding. Rebleeding is more likely to occur if the patient has hematemesis, liver disease, coagulopathy, hypotension and or anemia. There is a great need for conducting therapeutic trials as well as identifying predictors of outcome of upper GI bleeding in children to develop evidence based management protocols.


Adhesion of Fibroblasts to Immobilized alpha(2)-Macroglobulin.
This study examines effects of alpha(2)-macroglobulin (alpha(2)M) on adhesion of fibroblasts. Native alpha(2)M and transformed form of alpha(2)M, alpha(2)M-plasmin, were bound to plastic. Adhesion of mouse L929 and human embryo M-19 fibroblasts to immobilized alpha(2)M was estimated under various conditions by counting adherent cells using videomicroscopy and computer-assisted image analysis. alpha(2)M-plasmin, bound to plastic, induced adhesion and spreading of mouse L929 and human M-19 fibroblasts. Neither native alpha(2)M nor plasmin alone did not induce fibroblast adhesion. The adhesion to alpha(2)M-plasmin was undetectable at 4 degrees C, as well as when sodium azide was added or divalent ions were removed. These findings provide novel information on alpha(2)M functions. On the basis of these observations we hypothesized that alpha(2)M, immobilized in the extracellular matrix, can participate in the regulation of microenvironment effects on the cells, and, in particular, influence on fibroblast adhesion.


Pregnancy outcomes in patients with structural heart disease: a single center experience.
In women, pregnancy is a period of relatively drastic hemodynamic change in a short period of time. Most pregnant women adapt well to these gradual hemodynamic changes. However, in women with congenital heart disease or other structural heart disease, adaptation to theses sudden hemodynamic changes is difficult, and heart failure or arrhythmia can get aggravated. This study shares our experiences on the outcomes of pregnancy in patients with structural heart disease. From January 2007 to December 2016, we reviewed the medical records of all pregnant women with structural heart disease who received obstetric care at the Sejong General Hospital. During the study period, 103 pregnancies were observed in 79 women with structural heart disease. Of the 103 pregnancies, 55 were primiparous and 48 were multiparous. Echocardiography performed before pregnancy revealed that 52 patients had moderate to severe valvular regurgitation and 38 patients had moderate to severe valvular stenosis; 22 patients had mechanical valves and 5 patients had pulmonary hypertension. Overall, there were 9 maternal cardiac events, 7 obstetric events and 19 neonatal events. Pulmonary embolic events occurred only in 1 case; 77 deliveries were made, and 26 pregnancies did not last. Among 77 deliveries, 55 patients delivered by cesarean section (C/S) (71.43%). C/S in 16 of 55 patients was performed due to the maternal hemodynamic risk. Overall complications associated with pregnancy in women with structural heart disease were very high at 28.16%. However, it is hoped that maternal and neonatal outcomes will be improved through careful observation and preparedness for anticipated complications.


Oral administration of royal jelly facilitates mRNA expression of glial cell line-derived neurotrophic factor and neurofilament H in the hippocampus of the adult mouse brain.
Royal jelly (RJ) is known to have a variety of biological activities toward various types of cells and tissues of animal models, but nothing is known about its effect on brain functions. Hence, we examined the effect of oral administration of RJ on the mRNA expression of various neurotrophic factors, their receptors, and neural cell markers in the mouse brain. Our results revealed that RJ selectively facilitates the mRNA expression of glial cell line-derived neurotrophic factor (GDNF), a potent neurotrophic factor acting in the brain, and neurofilament H, a specific marker predominantly found in neuronal axons, in the adult mouse hippocampus. These observations suggest that RJ shows neurotrophic effects on the mature brain via stimulation of GDNF production, and that enhanced expression of neurofilament H mRNA is involved in events subsequently caused by GDNF. RJ may play neurotrophic and/or neuroprotective roles in the adult brain through GDNF.


Relationships between subjective or objective symptoms and mortality in schizophrenia: a prospective study on 310 schizophrenic patients with a median follow-up of 8.4 years.
The aim of the study was to explore the relationships between subjective or objective symptoms and mortality in schizophrenia. 310 subjects meeting the ICD-10 criteria for schizophrenia were included in the study between 1998 and 2000. At the initial assessment the following variables were respectively assessed to evaluate subjective and objective symptoms: the Frankfurt Complaints Questionnaire (FCQ) and the Positive and Negative Syndrome Scale (PANSS). In May 2008, information about the subjects were collected in order to know if they are alive or not and if they are deceased to know the date and the causes of their death. Survival analysis was conducted using the Kaplan-Meier product-limit estimator and standardized mortality ratio (SMR) was calculated. A multivariate Cox regression was done to detect predictive factors associated with mortality. Absolute mortality rates were 10.01%, 4.46% and 5.42% for overall mortality, unnatural causes and natural causes, respectively. SMR for overall mortality was 4.73. Cox regression analyses showed that elevated scores of FCQ was significant predictor of deaths from unnatural causes. High levels of subjective symptoms, as rated by the FCQ were independent predictor of mortality by unnatural causes in schizophrenic subjects. There were several limitations: The causes of death were not determined by autopsy and secondly, the duration of the study could be insufficient to detect significant associations between clinical variables and mortality.


A further approach to quantitative determinations by thin layer chromatography.
In the proposed method, an internal standard, suitably chosen, is added in equal quantities to the sample was well as to the standard solutions. A standard curve is obtained by plotting the ratio Ds/Di for each of the standard solutions against the logarithm of the quantity of material present in each solution, where Ds and Di are the average diameters of the standard and internal standard spots, respectively. It was disclosed that the ratio Ds/Di for a given solution is constant, regardless of the volume of the solution spotted. As a result, no exact equal known volumes are needed for spotting. The method has been tested with estimation of menthol in volatile oil of peppermint and strychine in nux vomica seed.


Immunological and histological study of temporal arteries.
Sixty-four temporal arteries were studied. 36 were from patients with clinically active temporal arteritis or polymyalgia rheumatica; 22 showed histological changes of temporal arteritis, 12 of which were in an active stage. 28 arteries, none of which showed histological changes, were taken at necropsy or from patients with unrelated disease. Extracellular immunoglobulin and complement deposition was seen in the artery biopsies showing active arteries and in 1 of the 10 biopsies with inactive arteritis. There was no immunoglobulin or complement deposition in the 14 patients with clinically active temporal arteritis and/or polymyalgia rheumatica, but with a normal artery biopsy. Patients with clinically active temporal arteritis were more likely to have a positive biopsy. Our results support the suggestion that the immune deposition is concurrent with an active histologically proven arteritis. Immunofluorescent examination does not appear to be a better diagnostic test than histological examination.


A new species of the genus Alterosa (Trichoptera: Philopotaminae) from Southeastern Brazil.
Philopotamidae is a large caddisfly family with 20 extant genera comprising about 1000 described species worldwide. Alterosa contains 22 described species distributed in South and Southeastern Brazil. Adults of the genus are very similar in general structure, but are remarkable for their wide variation in male genitalic morphology. The specimens were collected with Pennsylvania light traps and preserved in 80% ethanol. Alterosa nessimiani sp. nov. is described and figured from male specimens collected in streams of Rio de Janeiro state, Southeastern Brazil. The new species is closely related to A. jordaensis Blahnik 2005, from which it can be distinguished by the presence of two spines on the intermediate appendages and by the shape of tergum X.


Multiple desmoplastic cellular neurothekeomas localized to the face of a 16-year-old boy.
Cellular neurothekeomas are relative uncommon benign dermal tumors of uncertain histogenesis. Most commonly they arise as a solitary papule or nodule on the head and neck or upper trunk of young adults with a slight female predominance. There has been only 1 previous report of multiple neurothekeomas. The patient described herein was a 16-year-old otherwise healthy boy who presented with approximately 30 facial papules that arose over the course of 6 months and progressively enlarged. Histologically, all lesions were composed of spindled to epithelioid cells, but varied in the degree of cellularity and dermal sclerosis. Immunohistochemical staining showed that the cells of interest expressed S100A6, vimentin, CD63 (NKI/C3), PGP 9.5, and factor XIIIa and were negative for CD68, glial fibrillary acid protein (GFAP), S-100, HMB-45, epithelial membrane antigen, actin, and CD57 consistent with a diagnosis of multiple desmoplastic cellular neurothekeomas.


Genitopatellar syndrome in an adolescent female with severe osteoporosis and endocrine abnormalities.
Genitopatellar syndrome (GPS) is a rare disorder with characteristic craniofacial features, congenital flexion contractures of the lower limbs, absent or abnormal patellae, urogenital anomalies, and severe psychomotor retardation. Twelve patients with ages from 15 days to 12 years and two affected fetuses have been reported. We describe a 17-year-old female with a phenotype consistent with GPS. Being the oldest reported patient, she is the first one showing severe symptomatic osteoporosis and endocrine abnormalities including primary hypothyroidism and delayed puberty. We suggest that these novel findings are also manifestations of GPS.


Cognition and community functioning in schizophrenia: The nature of the relationship.
Although cognition is one of the most important predictors of community functioning in schizophrenia, little is known about the causes of this correlation. To our knowledge, this study is the first to examine the extent to which this correlation is genetically mediated and whether the genetic correlation is specific to schizophrenia. Six hundred thirty-six participants from 43 multigenerational families with at least two relatives with schizophrenia and 135 unrelated controls underwent diagnostic interview and cognition and functioning assessment. Quantitative genetic analyses were conducted using maximum-likelihood variance decomposition methods implemented in SOLAR (Almasy & Blangero, 1998). Among patients with schizophrenia, cognition and community functioning were positively correlated and genetic effects shared between them were significant contributors to this relationship whereas environmental effects shared between them were not. In contrast, genetic effects were not shared significantly between cognition in depressed or nondiagnosed relatives and community functioning in schizophrenia. In all analyses, the contributions of social cognition to community functioning were accounted for by general cognition. These findings support heritable factors that contribute to the correlation between cognition and community functioning that are relatively specific to schizophrenia and are not significantly shared with depression or a lack of psychopathology. This suggests the possibility of identifying specific genetic variants that contribute to this correlation and to these important individual differences among schizophrenia patients. (PsycINFO Database Record


Neutrophil extracellular traps in sheep mastitis.
Neutrophil extracellular traps (NETs) are structures composed of DNA, histones, and antimicrobial proteins that are released extracellularly by neutrophils and other immune cells as a means for trapping and killing invading pathogens. Here, we describe NET formation in milk and in mammary alveoli of mastitic sheep, and provide a dataset of proteins found in association to these structures. Nucleic acid staining, immunomicroscopy and fluorescent in-situ hybridization of mastitic mammary tissue from sheep infected with Streptococcus uberis demonstrated the presence of extranuclear DNA colocalizing with antimicrobial proteins, histones, and bacteria. Then, proteomic analysis by LTQ-Orbitrap Velos mass spectrometry provided detailed information on protein abundance changes occurring in milk upon infection. As a result, 1095 unique proteins were identified, of which 287 being significantly more abundant in mastitic milk. Upon protein ontology classification, the most represented localization classes for upregulated proteins were the cytoplasmic granule, the nucleus, and the mitochondrion, while function classes were mostly related to immune defence and inflammation pathways. All known NET markers were massively increased, including histones, granule proteases, and antimicrobial proteins. Of note was the detection of protein arginine deiminases (PAD3 and PAD4). These enzymes are responsible for citrullination, the post-translational modification that is known to trigger NET formation by inducing chromatin decondensation and extracellular release of NETs. As a further observation, citrullinated residues were detected by tandem mass spectrometry in histones of samples from mastitic animals. In conclusion, this work provides novel microscopic and proteomic information on NETs formed in vivo in the mammary gland, and reports the most complete database of proteins increased in milk upon bacterial mastitis.


LncRNA MIR217HG aggravates pressure-overload induced cardiac remodeling by activating miR-138/THBS1 pathway.
Cardiac hypertrophy and fibrosis are associated with cardiac remodeling and heart failure. We have previously shown that miRNA-217, embedded within the third intron of MIR217HG, aggravates pressure overload-induced cardiac hypertrophy by targeting phosphatase and tensin homolog. However, whether the MIR217HG transcript itself plays a role in cardiac remodeling remains unknown. Real-time PCR assays and RNA in situ hybridization were performed to detect MIR217HG expression. Lentiviruses and adeno-associated viruses with a cardiac-specific promoter (cTnT) were used to control MIR217HG expression in vitro and in vivo. Transverse aortic constriction (TAC) surgery was performed to develop cardiac remodeling models. Cardiac structure and function were analyzed using echocardiography and invasive pressure-volume analysis. MIR217HG expression was increased in patients with heart failure. MIR217HG overexpression aggravated pressure-overload-induced myocyte hypertrophy and fibrosis both in vivo and in vitro, whereas MIR217HG knockdown reversed these phenotypes. Mechanistically, MIR217HG increased THBS1 expression by sponging miR-138. MiR-138 recognized the 3'UTR of THBS1 and repressed THBS1 expression in the absence of MIR217HG. Silencing THBS1 expression reversed MIR217HG-induced cardiac hypertrophy and remodeling. MIR217HG acts as a potent inducer of cardiac remodeling that may contribute to heart failure by activating the miR-138/THBS1 pathway.


Risk factors for diabetic kidney disease in adults with longstanding type 1 diabetes: results from the Canadian Study of Longevity in Diabetes.
Objectives: Diabetic kidney disease (DKD) is an independent predictor of cardiovascular morbidity and mortality in type 1 diabetes (T1D). We aimed to explore clinical and biochemical factors, including the achievement of American Diabetes Association (ADA) recommended targets associated with DKD in people living with T1D for ≥50 years. Methods: This was a post hoc analysis of a cross-sectional study of 75 participants enrolled in the Canadian Study of Longevity in T1D. We explored diabetes-related complications, including neuropathy, retinopathy, cardiovascular disease, and DKD. Study participants were dichotomized based on the achievement of ADA recommended targets as the low-target group (achieving ≤4 targets, n = 31) and high-target group (achieving >4 targets, n = 44). The outcome of interest was DKD defined by estimated glomerular filtration rate (eGFR) values <60/mL/min/1.73 m2 and/or 24-h albumin excretion >30 mg. Multivariable logistic regression models were employed to estimate odds ratios (ORs) for DKD with 95% confidence intervals (CIs). Results: Of the 75 participants with prolonged T1D duration (45% male, mean age 66 years), 25 participants had DKD and 50 did not. There was no statistical difference between the high- and low-target groups in terms of age and body mass index. eGFR was significantly higher and the prevalence of diabetic retinopathy was significantly lower in the high-target group. Older age at diagnosis of T1D and lower frequency component to high-frequency component ratio increased the odds of having DKD. Conclusions: In adults with prolonged T1D duration, older age at diagnosis and lower heart rate variability may be associated with DKD.


Development of tridentate iron chelators: from desferrithiocin to ICL670.
Successful treatment of beta-thalassemia requires two key elements: blood transfusion and iron chelation. Regular blood transfusions considerably expand the lifespan of patients, however, without the removal of the consequential accumulation of body iron, few patients live beyond their second decade. In 1963, the introduction of desferrioxamine (DFO), a hexadentate chelator, marked a breakthrough in the treatment of beta-thalassemia. DFO significantly reduces body iron burden and iron-related morbidity and mortality. DFO is still the only drug for general use in the treatment of transfusion dependent iron overload. However, its very short plasma half-life and poor oral activity necessitate special modes of application (subcutaneous or intravenous infusion) which are inconvenient, can cause local reactions and are difficult to be accepted by many patients. Over the past four decades, many different laboratories have invested major efforts in the identification of orally active iron chelators from several hundreds of molecules of synthetic, microbial or plant origin. The discovery of ferrithiocin in 1980, followed by the synthesis of the tridentate chelator desferrithiocin and proof of its oral activity raised a lot of hope. However, the compound proved to be toxic in animals. Over a period of about fifteen years many desferrithiocin derivatives and molecules with broader alterations led to the discovery of numerous new compounds some of which were much better tolerated and were more efficacious than desferrithiocin in animals, however, none was safe enough to proceed to the clinical use. The discovery of a new chemical class of iron chelators: The bis-hydroxyphenyltriazoles re-energized the search for a safe tridentate chelator. The basic structure of this completely new chemical class of iron chelators was discovered by a combination of rational design, intuition and experience. More than forty derivatives of the triazole series were synthesized at Novartis. These compounds were evaluated, together with more than 700 chelators from various chemical classes. Using vigorous selection criteria with a focus on tolerability, the tridentate chelator 4-[(3,5-Bis-(2-hydroxyphenyl)-1,2,4)triazol-1-yl]-benzoic acid (ICL670) emerged as an entity which best combined high oral potency and tolerability in animals. ICL670 is presently being evaluated in the clinic.


Does the location of short-arm cast univalve effect pressure of the three-point mould?
Forearm and distal radius fractures are among the most common fractures in children. Many fractures are definitively treated with closed reduction and casting, however, the risk for re-displacement is high (7% to 39%). Proper cast application and the three-point moulding technique are modifiable factors that improve the ability of a cast to maintain the fracture reduction. Many providers univalve the cast to accommodate swelling. This study describes how the location of the univalve cut impacts the pressure at three-point mould sites for a typical dorsally displaced distal radius fracture. We placed nine force-sensing resistors on an arm model to collect pressure data at the three-point mould sites. Sensory inputs were sampled at 15 Hz. Cast padding and a three-point moulded short arm fibreglass cast was applied. The cast was then univalved on the dorsal, volar, radial or ulnar aspect. Pressure recordings were obtained throughout the procedure. A total of 24 casts were analyzed. Casts univalved in the sagittal plane (dorsal or volar surface) retained up to 16% more pressure across the three moulding sites compared with casts univalved in the coronal plane (radial or ulnar border). Maintaining pressure at the three-point mould prevents loss of reduction at the fracture site. This study shows that univalving the cast dorsally or volarly results in less pressure loss at moulding sites. This should improve the chances of maintaining fracture reductions when compared with radial or ulnar cuts in the cast. Sagittal plane univalving of forearm casts is recommended.


Epidemiology, responses, and way forward: the silent epidemic of viral hepatitis and HIV coinfection in Vietnam.
Viral hepatitis is a disease of great concern to public health that is now met by a favorable momentum to combat the global epidemic. This article is intended to highlight the importance of viral hepatitis in the Vietnam population as well in the group of people living with HIV/AIDS (PLWHA). We reviewed available data on epidemiology and response on hepatitis B virus (HBV) and hepatitis C virus (HCV) and HIV coinfection in Vietnam. The hepatitis B surface antigen (HBsAg) prevalence in the general population ranged from 5.7% to 24.7%. The anti-HCV prevalence ranged from 0.38% to 4.3% in the general population, while among people who inject drugs (PWIDs) it ranged from 31% to 97.2%. The HBV prevalence among PLWHA is similar to the general population, while HCV/HIV coinfection is concentrated in some groups. Anti-HCV prevalence among HIV-infected PWIDs can be as high as 98.5%. Developing policies for diagnosis and treatment of chronic HBV and HCV infections are critical priorities in order to prevent clinical progression to cirrhosis and liver cancer.


Use of the Herb Gymnema sylvestre to Illustrate the Principles of Gustatory Sensation: An Undergraduate Neuroscience Laboratory Exercise.
The Indian herb Gymnema sylvestre has been used in traditional Ayurvedic medicine for 2000 years, most recently for the treatment of diabetes. Loose leaf Gymnema sylvestre can be prepared as a tea and will impair the ability to taste sugar by blocking sweet receptors on the tongue. This report describes a laboratory exercise easily applied to an undergraduate neuroscience course that can be used to illustrate the principles of gustatory sensation. Combined with a preceding lecture on the primary taste sensations, students experience and appreciate how the primary tastes are combined to produce overall taste. In addition, the exercises outlined here expand upon previously published demonstrations employing Gymnema sylvestre to include illustrations of the different sensory transduction mechanisms associated with each of the four or five primary taste modalities. Students compare their qualitative primary taste experiences to salt, sugar, aspartame, chocolate, and sweet-sour candy prior to and following exposure to Gymnema sylvestre. The herb's impairment of sweet sensation is profound and dramatically alters the perception of sweetness in sugar, chocolate, and candy without altering the perception of the other primary tastes. The exercise has an indelible effect on students because the herb's intense effect compels students to rely on their unique personal experiences to highlight the principles of gustatory sensation.


Fingolimod: Assay analysis of US generic capsule products reveals variation in fingolimod content beyond the recommended acceptance criteria.
The immunomodulating agent fingolimod is a sphingosine-1-phosphate receptor modulator used in the treatment of multiple sclerosis (MS). We analyzed three FDA approved fingolimod 0.5 mg generic capsule products for fingolimod content. Assay results demonstrated a wide variation in fingolimod content between manufacturers, with one product demonstrating a fingolimod content of 76.8 % of the approved dose. This falls significantly below the FDA acceptance criteria of 90.0-110.0 % of label claim.