Anti-inflammatory activity of chloroquine and amodiaquine through p21-mediated suppression of T cell proliferation and Th1 cell differentiation.
Chloroquine (CQ) and amodiaquine (AQ) have been used for treating or preventing malaria for decades, and their application has expanded into treating inflammatory disease in humans. CQ and AQ are applicable for controlling rheumatoid arthritis, but their molecular mechanisms of anti-inflammatory activity remain to be elucidated. In this study, we examined the effects of CQ and AQ on T cell activation and T cell-mediated immune response. CQ had no significant effect on T cell numbers, but decreased the population of T cells with a high division rate. However, AQ treatment significantly increased the number of cells with low division rates and eliminated cells with high division rates, resulting in the inhibition of T cell proliferation triggered by T cell receptor stimulation, of which inhibition occurred in developing effector T helper and regulatory T cells, regardless of the different exogenous cytokines. Interestingly, the cyclin-dependent kinase inhibitor p21 was significantly and dose-dependently increased by CQ, and more potently by AQ, while other cell cycle regulators were unchanged. Both CQ and AQ elevated the transcription level of p21 though the activation of p53, but also blocked p21 protein degradation in the presence of cycloheximide, causing p21 protein accumulation mainly in the nucleus. Sustained treatment of developing T cells with either CQ or AQ suppressed IFN-γ production in a dose dependent manner and potently inhibited the differentiation of IFN-γ-producing Th1 cells. These results demonstrate that CQ and AQ increase the expression level of p21 and inhibit T cell proliferation and the development of IFN-γ-producing Th1 cells, thereby revealing beneficial roles in treating a wide range of chronic inflammatory diseases mediated by inflammatory T cells.


Evaluation of transanal hemorrhoidal dearterialization: a single surgeon experience.
There is an increasing, though still limited, amount of evidence describing the use of the transanal hemorrhoidal dearterialization (THD) device for the treatment of hemorrhoidal disease. This study assesses postoperative outcomes from a single surgeon experience with the THD device. From January 2009 to December 2011, 108 THD procedures were performed. With Doppler guidance, the THD device makes possible precise ligation of the branches of the superior hemorrhoidal artery. Patients were seen postoperatively at 3 weeks and 6 months. They underwent physical examination to determine whether there was recurrence of hemorrhoidal prolapse. They were asked to describe any bleeding, to rate pain using the visual analog scale, and to rate their level of satisfaction on a scale of 1-5 (with 5 = highly satisfied). A phone interview was used for follow-up at 1 year to determine the rate of recurrent prolapse. Of the 108 patients who underwent THD, two were lost to follow-up and excluded. All of the remaining 106 patients completed follow-up at 3 weeks and 6 months. At 3 weeks, 92% of patients had no pain and 88% were highly satisfied with the procedure at 3 weeks. This increased to 92% satisfaction at 1 year. Prolapse recurrence was 7.5% at 6 months and 10.3% at 1 year. Bleeding was the most common complication, but did not require re-intervention or transfusion. THD is a same-day procedure for the treatment of hemorrhoidal disease that is safe and effective, and offers the potential for immediate return to normal activity.


Hypersynchronization in mild cognitive impairment: the 'X' model.
Hypersynchronization has been proposed as a synaptic dysfunction biomarker in the Alzheimer's disease continuum, reflecting the alteration of the excitation/inhibition balance. While animal models have verified this idea extensively, there is still no clear evidence in humans. Here we test this hypothesis, evaluating the risk of conversion from mild cognitive impairment (MCI) to Alzheimer's disease in a longitudinal study. We compared the functional resting state eyes-closed magnetoencephalographic networks of 54 patients with MCI who were followed-up every 6 months. According to their clinical outcome, they were split into: (i) the 'progressive' MCI (n = 27) group; and (ii) the 'stable' MCI group (n = 27). They did not differ in gender or educational level. For all participants, two magnetoencephalographic recordings were acquired. Functional connectivity was evaluated using the phase locking value. To extract the functional connectivity network with significant changes between both magnetoencephalographic recordings, we evaluated the functional connectivity ratio, defined as functional connectivity post-/pre-condition, in a network-based statistical model with an ANCOVA test with age as covariate. Two significant networks were found in the theta and beta bands, involving fronto-temporal and fronto-occipital connections, and showing a diminished functional connectivity ratio in the progressive MCI group. These topologies were then evaluated at each condition showing that at baseline, patients with progressive MCI showed higher synchronization than patients with stable MCI, while in the post-condition this pattern was reversed. These results may be influenced by two main factors in the post-condition: the increased synchrony in the stable MCI patients and the network failure in the progressive MCI patients. These findings may be explained as an 'X' form model where the hypersynchrony predicts conversion, leading subsequently to a network breakdown in progressive MCI. Patients with stable MCI showed an opposite phenomenon, which could indicate that they were a step beyond in the Alzheimer's disease continuum. This model would be able to predict the risk for the conversion to dementia in MCI patients.


Glucuronides in anti-cancer therapy.
Glucuronide prodrugs have shown promising efficacy in anti-cancer therapy due to their increased specificity and reduced systemic toxicity. The prodrugs can be used in prodrug monotherapy (PMT), which is based on elevated tumor beta-glucuronidase activity. beta-Glucuronidase activates the low-toxic prodrugs into highly cytotoxic agents specifically in the tumor site. The specificity of the prodrugs can be further improved by combined use with monoclonal antibodies against tumor-specific antigens, namely antibody-directed enzyme prodrug therapy (ADEPT); and the potency of the prodrugs can be greatly enhanced with the incorporation of an appropriate radionuclide in the combined chemo- and radio-therapy of cancer (CCRTC) strategy. The prodrugs can also be utilized to modify liposomes for efficient delivery of anti-cancer drugs.


Activation of elements in HERV-W family by caffeine and aspirin.
Caffeine and aspirin have been suggested to be involved in neurologic diseases, such as schizophrenia, and previous data have revealed that abnormal expression of HERV-W elements may be an important factor in the etiopathogenesis of those diseases. In this article, we reported that caffeine and aspirin contributed to the expression of HERV-W env and gag in Human SH-SY5Y neuroblastoma cells. Semi-quantitative RT-PCR and quantitative Real-time PCR were used to detect the mRNA of HERV-W env and gag in cells exposed to caffeine or aspirin. Western blotting was used to detect the protein of HERV-W env. Luciferase activity assay was employed to detect the activity of HERV-W env promoter. It was found that both caffeine and aspirin could increase the expression of HERV-W env and gag in human SH-SY5Y neuroblastoma cells. Caffeine could activate the HERV-W env promoter, while aspirin could not. With previous studies we can conjecture that HERVs might play a bridging role between environmental factors, such as drugs and neurologic diseases.


Stress-Related Disorders of Family Members of Patients Admitted to the Intensive Care Unit With COVID-19.
The psychological symptoms associated with having a family member admitted to the intensive care unit (ICU) during the COVID-19 pandemic are not well defined. To examine the prevalence of symptoms of stress-related disorders, primarily posttraumatic stress disorder (PTSD), in family members of patients admitted to the ICU with COVID-19 approximately 90 days after admission. This prospective, multisite, mixed-methods observational cohort study assessed 330 family members of patients admitted to the ICU (except in New York City, which had a random sample of 25% of all admitted patients per month) between February 1 and July 31, 2020, at 8 academic-affiliated and 4 community-based hospitals in 5 US states. Having a family member in the ICU with COVID-19. Symptoms of PTSD at 3 months, as defined by a score of 10 or higher on the Impact of Events Scale 6 (IES-6). A total of 330 participants (mean [SD] age, 51.2 [15.1] years; 228 [69.1%] women; 150 [52.8%] White; 92 [29.8%] Hispanic) were surveyed at the 3-month time point. Most individuals were the patients' child (129 [40.6%]) or spouse or partner (81 [25.5%]). The mean (SD) IES-6 score at 3 months was 11.9 (6.1), with 201 of 316 respondents (63.6%) having scores of 10 or higher, indicating significant symptoms of PTSD. Female participants had an adjusted mean IES-6 score of 2.6 points higher (95% CI, 1.4-3.8; P < .001) than male participants, whereas Hispanic participants scored a mean of 2.7 points higher compared with non-Hispanic participants (95% CI, 1.0-4.3; P = .002). Those with graduate school experience had an adjusted mean score of 3.3 points lower (95% CI, 1.5-5.1; P < .001) compared with those with up to a high school degree or equivalent. Qualitative analyses found no substantive differences in the emotional or communication-related experiences between those with high vs low PTSD scores, but those with higher scores exhibited more distrust of practitioners. In this cohort study, symptoms of PTSD among family members of ICU patients with COVID-19 were high. Hispanic ethnicity and female gender were associated with higher symptoms. Those with higher scores reported more distrust of practitioners.


Gα12 structural determinants of Hsp90 interaction are necessary for serum response element-mediated transcriptional activation.
The G12/13 class of heterotrimeric G proteins, comprising the α-subunits Gα12 and Gα13, regulates multiple aspects of cellular behavior, including proliferation and cytoskeletal rearrangements. Although guanine nucleotide exchange factors for the monomeric G protein Rho (RhoGEFs) are well characterized as effectors of this G protein class, a variety of other downstream targets has been reported. To identify Gα12 determinants that mediate specific protein interactions, we used a structural and evolutionary comparison between the G12/13, Gs, Gi, and Gq classes to identify "class-distinctive" residues in Gα12 and Gα13. Mutation of these residues in Gα12 to their deduced ancestral forms revealed a subset necessary for activation of serum response element (SRE)-mediated transcription, a G12/13-stimulated pathway implicated in cell proliferative signaling. Unexpectedly, this subset of Gα12 mutants showed impaired binding to heat-shock protein 90 (Hsp90) while retaining binding to RhoGEFs. Corresponding mutants of Gα13 exhibited robust SRE activation, suggesting a Gα12-specific mechanism, and inhibition of Hsp90 by geldanamycin or small interfering RNA-mediated lowering of Hsp90 levels resulted in greater downregulation of Gα12 than Gα13 signaling in SRE activation experiments. Furthermore, the Drosophila G12/13 homolog Concertina was unable to signal to SRE in mammalian cells, and Gα12:Concertina chimeras revealed Gα12-specific determinants of SRE activation within the switch regions and a C-terminal region. These findings identify Gα12 determinants of SRE activation, implicate Gα12:Hsp90 interaction in this signaling mechanism, and illuminate structural features that arose during evolution of Gα12 and Gα13 to allow bifurcated mechanisms of signaling to a common cell proliferative pathway.


Identification of a conserved var gene in different Plasmodium falciparum strains.
The multicopy var gene family of Plasmodium falciparum is of crucial importance for pathogenesis and antigenic variation. So far only var2csa, the var gene responsible for placental malaria, was found to be highly conserved among all P. falciparum strains. Here, a new conserved 3D7 var gene (PF3D7_0617400) is identified in several field isolates. DNA sequencing, transcriptional analysis, Cluster of Differentiation (CD) 36-receptor binding, indirect immunofluorescence with PF3D7_0617400-antibodies and quantification of surface reactivity against semi-immune sera were used to characterize an NF54 clone and a Gabonese field isolate clone (MOA C3) transcribing the gene. A population of 714 whole genome sequenced parasites was analysed to characterize the conservation of the locus in African and Asian isolates. The genetic diversity of two var2csa fragments was compared with the genetic diversity of 57 microsatellites fragments in field isolates. PFGA01_060022400 was identified in a Gabonese parasite isolate (MOA) from a chronic infection and found to be 99% identical with PF3D7_0617400 of the 3D7 genome strain. Transcriptional analysis and immunofluorescence showed expression of the gene in an NF54 and a MOA clone but CD36 binding assays and surface reactivity to semi-immune sera differed markedly in the two clones. Long-read Pacific bioscience whole genome sequencing showed that PFGA01_060022400 is located in the internal cluster of chromosome 6. The full length PFGA01_060022400 was detected in 36 of 714 P. falciparum isolates and 500 bp fragments were identified in more than 100 isolates. var2csa was in parts highly conserved (He = 0) but in other parts as variable (He = 0.86) as the 57 microsatellites markers (He = 0.8). Individual var gene sequences exhibit conservation in the global parasite population suggesting that purifying selection may limit overall genetic diversity of some var genes. Notably, field and laboratory isolates expressing the same var gene exhibit markedly different phenotypes.


Raman-microscopy investigation of vitrification-induced structural damages in mature bovine oocytes.
Although oocyte cryopreservation has great potentials in the field of reproductive technologies, it still is an open challenge in the majority of domestic animals and little is known on the biochemical transformation induced by this process in the different cellular compartments. Raman micro-spectroscopy allows the non-invasive evaluation of the molecular composition of cells, based on the inelastic scattering of laser photons by vibrating molecules. The aim of this work was to assess the biochemical modifications of both the zona pellucida and cytoplasm of vitrified/warmed in vitro matured bovine oocytes at different post-warming times. By taking advantage of Principal Component Analysis, we were able to shed light on the biochemical transformation induced by the cryogenic treatment, also pointing out the specific role of cryoprotective agents (CPs). Our results suggest that vitrification induces a transformation of the protein secondary structure from the α-helices to the β-sheet form, while lipids tend to assume a more packed configuration in the zona pellucida. Both modifications result in a mechanical hardening of this cellular compartment, which could account for the reduced fertility rates of vitrified oocytes. Furthermore, biochemical modifications were observed at the cytoplasmic level in the protein secondary structure, with α-helices loss, suggesting cold protein denaturation. In addition, a decrease of lipid unsaturation was found in vitrified oocytes, suggesting oxidative damages. Interestingly, most modifications were not observed in oocytes exposed to CPs, suggesting that they do not severely affect the biochemical architecture of the oocyte. Nevertheless, in oocytes exposed to CPs decreased developmental competence and increased reactive oxygen species production were observed compared to the control. A more severe reduction of cleavage and blastocyst rates after in vitro fertilization was obtained from vitrified oocytes. Our experimental outcomes also suggest a certain degree of reversibility of the induced transformations, which renders vitrified oocytes more similar to untreated cells after 2 h warming.


Metabolic restaging of hepatocellular carcinoma using whole-body F-FDG PET/CT.
To evaluate the ability of (18)F-fluorodeoxyglucose positron emission and computed tomography ((18)F-FDG PET/CT) in restaging of hepatocellular carcinoma (HCC) after treatment. We reviewed a database of the diagnostic performance of (18)F-FDG PET/CT scan for patients with HCC following local or regional treatment. The database consisted of (18)F-FDG PET/CT information of 21 male and 4 female (age range, 27-81 years; mean age, 51.6 years) patients who had received surgical resection and/or interventional treatments and then underwent (18)F-FDG PET/CT scan. All patients had received enhanced CT scan of the liver two weeks before or after the (18)F-FDG PET/CT scan. Intrahepatic recurrence and/or extrahepatic metastases were confirmed by histological analysis or clinical and imaging follow-up. The accuracy of (18)F-FDG PET/CT study was determined by histopathological results or by clinical and imaging follow-up. (18)F-FDG PET/CT was abnormal in 19 of the 25 (76.0%) patients. In detecting HCC recurrence, (18)F-FDG PET/CT scored 17 true positives, 5 true negatives, 2 false positives and 1 false negative. The sensitivity, specificity and accuracy of (18)F-FDG PET/CT in detecting HCC recurrence was 89.5%, 83.3% and 88%, respectively. (18)F-FDG PET/CT had an impact on management of these patients by settling the problem of an unexplained increase in alpha-fetoprotein after treatment (14 patients), by monitoring response to the treatment and guiding additional regional therapy (12 patients), by identifying extrahepatic metastases (10 patients), by identifying tumor growth or thrombosis in the portal vein (6 patients), or by guiding surgical resection of extrahepatic metastases (2 patients). Our results suggest that whole body (18)F-FDG PET/CT may be useful in the early evaluation of residual, intrahepatic recurrent or extrahepatic metastatic lesions and able to provide valuable information for the management of HCC recurrence.


Screening of potent inhibitor from Aquilaria malaccensis Lam. against arachidonic inflammatory enzymes: an insight from molecular docking, ADMET, molecular dynamics simulation and MM-PBSA approaches.
The three primary enzymes COX (cyclooxygenase), LOX (lipoxygenase) and CYT-P450 (cytochrome P450), which are part of the arachidonic inflammatory pathway, play crucial role in the development of asthma, rheumatoid arthritis and cardiovascular diseases. Ethnomedicinally, plant-derived chemicals have a major role in the treatment of fatal illnesses. Aquilaria malaccensis Lam. widely known as agarwood is prized for its fragrance and therapeutic properties. The phytochemicals and extracts of this plant have significant healing properties in the treatment of serious illnesses. In the current work, an in-silico approach including molecular docking, ADMET (absorption, distribution, metabolism, excretion and toxicity), molecular dynamics (MD) simulation and molecular mechanics/Poisson-Boltzmann surface area (MM-PBSA) was performed to screen 33 bioactive compounds from this plant against COX-2 and 5-LOX in order to find the most effective inhibitor. 2-(2-Phenylethyl)chromone was found to inhibit both 5-LOX and COX-2, showing the highest binding affinities (-9.1 kcal/mol and -9.0 kcal/mol, respectively) than standard Ibuprofen and nordihydroguaiaretic acid (NDGA). 2-(2-Phenylethyl)chromone showed the highest drug-likeness score and low risk of toxicity compared to other phytochemicals. MD modeling and MM-PBSA calculations showed that 2-(2-Phenylethyl)chromone had a strong persistent binding interaction with 5-LOX than COX-2, and this interaction is comparable to the bounded standards Ibuprofen and NDGA. From this study, we may infer that the 2-(2-Phenylethyl)chromone can serve as a potent inhibitor and has scope to be employed in the treatment of inflammatory ailments.Communicated by Ramaswamy H. Sarma.


Oral omega-6 essential fatty acid treatment in contact lens associated dry eye.
Symptoms of dry eye are commonly reported in contact lens wearers and are a frequent cause of non-tolerance. The purpose of the present study is to evaluate the effects of oral treatment with particular omega-6 fatty acids in the form of evening primrose oil (EPO) on subjective symptoms, ocular surface signs and tear film characteristic in patients with contact lens associated dry eye. The study design was randomised, double-masked and placebo controlled. 76 female soft contact lens wearers were treated for 6 months with either EPO or placebo (olive oil). Subjects underwent three examinations (baseline, 3 and 6 months). At each examination subjects were given a questionnaire relating to lens comfort and dry eye symptoms and underwent a series of tests of tear film characteristics (tear meniscus height, break-up time), meibomian gland function (lipid layer thickness and quality) and ocular surface parameters (hyperaemia and staining). The EPO group showed a significant improvement in the specific symptom of 'dryness' at 3 and 6 months (p<0.01) and also a significant improvement in overall lens comfort at 6 months (p<0.01). Tear meniscus height was increased in the EPO group at 6 months relative to baseline (p<0.01), although all other objective signs were unchanged. This study provides evidence for a beneficial effect of particular orally administered omega-6 fatty acids in alleviating dry eye symptoms and improving overall lens comfort in patients suffering from contact lens associated dry eye.


Massage therapy for patients undergoing autologous bone marrow transplantation.
The purpose of the current study was to examine the impact of massage therapy on psychological, physical, and psychophysiological measures in patients undergoing autologous bone marrow transplantation (BMT). Patients scheduled to undergo BMT were randomly assigned to receive either (a) massage therapy, consisting of 20-minute sessions of shoulder, neck, head, and facial massage, or (b) standard treatment. Overall effects of massage therapy on anxiety, depression, and mood were assessed pretreatment, midtreatment, and prior to discharge using the State-Trait Anxiety Inventory, Beck Depression Inventory, and Brief Profile of Mood States, respectively. The immediate effects of massage were measured via the State Anxiety Inventory, Numerical Scales of Distress, Fatigue, Nausea, and Pain and indices of psychophysiological arousal (heart rate, blood pressure, and respiration rate), collected prior to and following patients' first, fifth, and final massage (on Days--7, midtreatment, and predischarge). Analysis of the data evaluating the immediate effects of massage showed that patients in the massage therapy group demonstrated significantly larger reductions in distress, fatigue, nausea, and State Anxiety than the standard treatment group at Day-7, in State Anxiety at midtreatment, and in fatigue at the predischarge assessment. The overall measures of psychological symptoms measured at pretreatment, midtreatment, and prior to discharge showed no overall group differences, although the massage group scored significantly lower on the State Anxiety Inventory than the standard care group at the midtreatment assessment. The two groups together showed significant declines through time on scores from the Profile of Mood States and State and Trait Anxiety Inventories.


CT and MR imaging of the thoracic aorta.
At present time, both CT and MRI are valuable techniques in the study of the thoracic aorta. Nowadays, CT represents the most widely employed technique for the study of the thoracic aorta. The new generation CTs show sensitivities up to 100% and specificities of 98-99%. Sixteen and wider row detectors provide isotropic pixels, mandatory for the ineludible longitudinal reconstruction. The main limits are related to the X-ray dose expoure and the use of iodinated contrast media. MRI has great potential in the study of the thoracic aorta. Nevertheless, if compared to CT, acquisition times remain longer and movement artifact susceptibility higher. The main MRI disadvantages are claustrophobia, presence of ferromagnetic implants, pacemakers, longer acquisition times with respect to CT, inability to use contrast media in cases of renal insufficiency, lower spatial resolution and less availability than CT. CT is preferred in the acute aortic disease. Nevertheless, since it requires iodinated contrast media and X-ray exposure, it may be adequately replaced by MRI in the follow up of aortic diseases. The main limitation of MRI, however, is related to the scarce visibility of stents and calcifications.


All-fiber fused directional coupler for highly efficient spatial mode conversion.
We model and demonstrate a simple mode selective all-fiber coupler capable of exciting specific higher order modes in two- and few-mode fibres with high efficiency and purity. The coupler is based on inter-modally phase-matching the propagation constants in each arm of the asymmetric fused coupler, formed by dissimilar fibres. At a specific coupler diameter, the launched fundamental LP(01) mode is coupled into the higher order mode (LP(11), LP(21), LP(02)) in the other arm, over a broadband wave-length range around 1550 nm. Unlike other techniques, the demonstrated coupler is composed of a multimode fiber that is weakly fused with a phase matched conventional single mode telecom fiber (SMF-28). The beating between the supermodes at the coupler waist produces a periodic power transfer between the two arms, and therefore, by monitoring the beating while tapering, it is possible to obtain optimum selection for the desired mode. High coupling efficiencies in excess of 90% for all the higher order modes were recorded over 100 nm spectral range, while insertion losses remain as low as 0.5 dB. Coupling efficiency can be further enhanced by performing slow tapering at high temperature, in order to precisely control the coupler cross-section geometry.


Serotonin noradrenaline reuptake inhibitors: New hope for the treatment of chronic pain.
Depression and painful symptoms occur frequently together. Over 75% of depressed patients report painful symptoms such as headache, stomach pain, neck and back pain as well as non-specific generalized pain. In addition, World Health Organization data have shown that primary care patients with chronic pain have a four fold greater risk of becoming depressed than pain-free patients. Increasingly, pain is considered as an integral symptom of depression and there evidence to suggest that pain and depression may arise from a common neurobiological dysfunction. Serotonergic cell bodies, in the raphe nucleus, and noradrenergic cell bodies in the locus coeruleus send projections to various parts of the brain, where they are involved in the control of mood, movement, cognitive functioning and emotions. In addition both serotonergic and noradrenergic neurons project to the spinal cord. These descending pathways serve to inhibit input from the intestines, skeletal muscles and other sensory inputs. Usually, these inhibitory effects are modest, but in times of stress, in the interest of the survival of the individual, they can completely inhibit the input from painful stimuli. A dysfunction of the serotonergic and noradrenergic neurons can thus affect both the ascending and descending pathways resulting in the psychological symptoms of depression and somatic pain symptoms such as chronic pain, fibromyalgia, non-cardiac chest pain, or irritable bowel syndrome. In view of this, it is not surprising that tricyclic antidepressants have been a standard treatment of chronic pain for many years. In contrast and in spite of their improved tolerance, selective serotonin reuptake inhibitors do not appear to be particularly effective in the treatment of pain. Recently, a number of open and controlled trials with selective serotonin and noradrenaline reuptake inhibitors such as venlafaxine, milnacipran and duloxetine, suggest that these compounds may be more effective in relieving pain than selective inhibitors of serotonin reuptake. Wherever valid comparisons have been made the newer dual action drugs appear to be as effective as the tricyclic and considerably better tolerated. Dual action antidepressants may thus soon become the new standard treatment of chronic pain whether it is associated with depression or not. In addition, these agents may also have a role in modulating neurogenesis and other neuroplastic changes in the central nervous system, thereby leading to more complete recovery in patients suffering from the symptoms of depression or chronic pain.


Cardiopulmonary Exercise Testing in Patients with Heart Failure with Specific Comorbidities.
Exercise capacity is one of the most powerful predicting factors of life expectancy, both in patients with and those without cardiac disease. Cardiopulmonary exercise testing provides a global assessment of the integrative exercise responses involving the pulmonary, cardiovascular, hematopoietic, neuropsychological, and skeletal muscle systems, which are not adequately reflected through the measurement of individual organ system function. This relatively noninvasive, dynamic, physiologic overview allows the evaluation of both submaximal and peak exercise responses, providing the physician with relevant information for clinical decision-making. Chronic heart failure is a significant cause of worldwide mortality and morbidity, whose clinical picture is characterized by exercise intolerance and impaired quality of life. The purpose of this review is to provide an update of the role of cardiopulmonary exercise testing in patients with heart failure with specific comorbidities. Patients with heart failure frequently present concomitant clinical conditions, such as obesity, anemia, lung or kidney disease, diabetes mellitus, cancer, depression, and psychogenic disorders, which could affect length and quality of life, including everyday activities and exercise performance. Poor effort and malingering may be suspected when early discontinuation of the exercise test with irregular breathing occurs.


Ile73Asn mutation in protein C introduces a new N-linked glycosylation site on the first EGF-domain of protein C and causes thrombosis.
Activated protein C exerts its anticoagulant activity by protein S-dependent inactivation of factors Va and VIIIa by limited proteolysis. We identified a venous thrombosis patient who has plasma protein C antigen level of 63% and activity levels of 44% and 23%, as monitored by chromogenic and clotting assays. Genetic analysis revealed the proband carries compound heterozygous mutations (c.344T>A, p.I73N and c.1181G>A, p.R352Q) in PROC We individually expressed protein C mutations and discovered that thrombin-thrombomodulin activates both variants normally and the resulting activated protein C mutants exhibit normal amidolytic and proteolytic activities. However, while protein S-dependent catalytic activity of activated protein C-R352Q toward factor Va was normal, it was significantly impaired for activated protein C-I73N. These results suggest that the Ile to Asn substitution impairs interaction of activated protein C-I73N with protein S. This conclusion was supported by a normal anticoagulant activity for activated protein C-I73N in protein S-deficient but not in normal plasma. Further analysis revealed Ile to Asn substitution introduces a new glycosylation site on first EGF-like domain of protein C, thereby adversely affecting interaction of activated protein C with protein S. Activated protein C-R352Q only exhibited reduced activity in sub-physiological concentrations of Na+ and Ca2+, suggesting that this residue contributes to metal ion-binding affinity of the protease, with no apparent adverse effect on its function in the presence of physiological levels of metal ions. These results provide insight into the mechanism by which I73N/R352Q mutations in activated protein C cause thrombosis in proband carrying this compound heterozygous mutation.


Activated terminal complement in cerebrospinal fluid in Guillain-Barré syndrome and multiple sclerosis.
A quantitative enzyme-linked immunosorbent assay was used to measure the concentration of fluid-phase complement C5b-9 complexes (SC5b-9) in the cerebrospinal fluid (CSF) of 14 patients with acute monophasic Guillain-Barré Syndrome (GBS), 21 patients with multiple sclerosis (MS), and 11 patients with noninflammatory central nervous system (CNS) diseases. SC5b-9 complexes were detected in the CSF of 13 of 14 patients with acute GBS (mean, 3.08 micrograms/ml; range, 0 to 7.1 micrograms/ml) and 16 of 21 patients with MS (mean, 1.83 micrograms/ml; range, 0 to 7.5 micrograms/ml). In the control group of patients with noninflammatory CNS diseases, SC5b-9 was not detected in eight of 11 and was present in low concentrations in the remaining three patients (mean, 0.28 micrograms/ml; range, 0 to 1.7 micrograms/ml). The finding of SC5b-9 complexes in the CSF of patients with GBS and MS suggests that terminal complement components may participate in the tissue-damaging processes in these diseases.


Does lorazepam impair the antidepressant response to nortriptyline and psychotherapy?
This analysis sought to determine whether lorazepam influences time to response or rate of response in elderly depressed patients receiving nortriptyline and psychotherapy and to examine clinical and polysomnographic correlates of lorazepam treatment. Patients with recurrent major depressive disorder (N = 119; mean +/- SD age = 68.0 +/- 6.1 years; diagnosis defined by Research Diagnostic Criteria) received acute treatment with nortriptyline and interpersonal psychotherapy. Thirty-five patients received open-label adjunctive lorazepam for anxiety or insomnia symptoms (LZ+) and 84 did not. Statistical analyses were conducted between the LZ+ group and a group of 35 patients who received no lorazepam (LZ-) and were matched for anxiety level. Patients had polysomnographic studies prior to treatment and after remission of depressive symptoms. The LZ+ group reported more anxiety on the Brief Symptom Inventory (p = .04) compared with the remaining 84 patients. The LZ+ group had a greater proportion of endogenous depression subtype than the anxiety-matched LZ- group, in addition to more abnormal EEG sleep (higher percentage of REM sleep, shorter REM latency, lower delta sleep ratio). Mean time to initial antidepressant response was no different between groups. However, a significantly greater proportion of LZ+ than LZ- patients responded to acute treatment (91.4% vs. 71.4%; p < .03). Adjunctive lorazepam does not slow the antidepressant response to combined antidepressant/psychotherapy treatment in elderly depressed patients, and it is associated with a greater likelihood of antidepressant response. A greater percentage of patients treated with lorazepam have endogenous depression subtype and abnormal sleep findings (EEG) than those who are not treated with lorazepam. Adjunctive lorazepam is useful for treating anxiety in elderly depressed patients.


Dendrimer-like alpha-d-glucan nanoparticles activate dendritic cells and are effective vaccine adjuvants.
The use of nanoparticles for delivery of vaccine antigens and as vaccine adjuvants is appealing because their size allows efficient uptake by dendritic cells and their biological properties can be tailored to the desired function. Here, we report the effect of chemically modified phytoglycogen, a dendrimer-like α-d-glucan nanoparticle, on dendritic cells in vitro, and the utility of this type of nanoparticle as a vaccine adjuvant in vivo. The modified phytoglycogen nanoparticle, termed Nano-11, has a positive surface charge which enabled electrostatic adsorption of negatively charged protein antigens. The Nano-11-antigen complexes were efficiently phagocytized by dendritic cells. Nano-11 induced increased expression of costimulatory molecules and the secretion of IL-1β and IL-12p40 by dendritic cells. Intramuscular injection of Nano-11-antigen formulations induced a significantly enhanced immune response to two different protein antigens. Examination of the injection site revealed numerous monocytes and relatively few neutrophils at one day after injection. The inflammation had nearly completely disappeared by 2 weeks after injection. These studies indicate that Nano-11 is an effective vaccine delivery vehicle that significantly enhances the immune response. This type of plant based nanoparticle is considered highly cost-effective compared with fully synthetic nanoparticles and appears to have an excellent safety profile making them an attractive adjuvant candidate for prophylactic vaccines.


An activation-specific role for transcription factor TFIIB in vivo.
A yeast mutant was isolated encoding a single amino acid substitution [serine-53 --> proline (S53P)] in transcription factor TFIIB that impairs activation of the PHO5 gene in response to phosphate starvation. This effect is activation-specific because S53P did not affect the uninduced level of PHO5 expression, yet is not specific to PHO5 because Adr1-mediated activation of the ADH2 gene also was impaired by S53P. Pho4, the principal activator of PHO5, directly interacted with TFIIB in vitro, and this interaction was impaired by the S53P replacement. Furthermore, Pho4 induced a conformational change in TFIIB, detected by enhanced sensitivity to V8 protease. The S53P replacement also impaired activation of a lexA(op)-lacZ reporter by a LexA fusion protein to the activation domain of Adr1, thereby indicating that the transcriptional effect on ADH2 expression is specific to the activation function of Adr1. These results define an activation-specific role for TFIIB in vivo and suggest that certain activators induce a conformational change in TFIIB as part of their mechanism of transcriptional stimulation.


Depression in dermatology: an integrative perspective.
An example which confirmes the adequacy of the integrative perspective in medicine is depression in dermatology because of the sufficient number of arguments linking depression and some chronic skin conditions in depth more than simply comorbidity. In recent literature, the authors found, how depression in dermatology occurs significantly more frequently than in the general population. In dermatological patients the prevalence of depression is around 30% which is more in comparison to patients in general practice where prevalence of depression is 22%. The authors found a considerable similarity in the various characteristics between depression and psoriasis, based upon recent research findings in immunity disorder and elevated concentrations of proinflammatory cytokines as well as acute phase proteins in both disorders, indicating that both disorders can be considered as immunologicaly mediated, inflammatory states with repetitive chronic progress and similar comorbidity. The afore mentioned theoretical settings evoke the integrative aspect and the integrative interdisciplinary approach placing in the center of attention not only the diseased person with his fears, needs, preocupations and expectations during treatment, but the person who is at risk of becoming burdened with these disorders.


Small artery compliance in cirrhotic patients during total paracentes.
Hemodynamic changes, including systemic vascular resistance, in cirrhotic patients during massive paracentesis have been reported, but large and small artery compliance has not yet been investigated. To investigate hemodynamic variables, including small and large artery compliance, in cirrhotic patients during total paracentesis. The study included 15 cirrhotic patients admitted for an episode of tense diuretic-resistant ascites. Hemodynamic variables including vascular compliance were measured using an HDI pulse wave cardiovascular profiling instrument CR-2000. The variables were measured in these patients before, immediately after, and 24 hours following large volume (mean 5.6 L) paracentesis. Cardiac output increased immediately after paracentesis due to increment in stroke volume, with no change in heart rate. However, 24 hours later the cardiac output decreased to below the basal level. The fluctuation was statistically significant (P < 0.05). There was no change in large artery compliance, but small artery compliance increased after paracentesis (P < 0.05) and partially retumed to the basal level after 24 hours. Systemic vascular resistance measurement showed the same pattern of change: vasodilatation occurred during paracentesis and was attenuated 24 hours later. Large volume paracentesis with albumin replacement caused an accentuation of the vasodilatation (small but not large artery) already present in these patients. This may be the first sign of enhanced vasodilatation due to large volume paracentesis before the clinical expression of impaired hemodynamics and deterioration of renal function.


Glypican 4 modulates FGF signalling and regulates dorsoventral forebrain patterning in Xenopus embryos.
Heparan sulphate proteoglycans such as glypicans are essential modulators of intercellular communication during embryogenesis. In Xenopus laevis embryos, the temporal and spatial distribution of Glypican 4 (Gpc4) transcripts during gastrulation and neurulation suggests functions in early development of the central nervous system. We have functionally analysed the role of Xenopus Gpc4 by using antisense morpholino oligonucleotides and show that Gpc4 is part of the signalling network that patterns the forebrain. Depletion of GPC4 protein results in a pleiotropic phenotype affecting both primary axis formation and early patterning of the anterior central nervous system. Molecular analysis shows that posterior axis elongation during gastrulation is affected in GPC4-depleted embryos, whereas head and neural induction are apparently normal. During neurulation, loss of GPC4 disrupts expression of dorsal forebrain genes, such as Emx2, whereas genes marking the ventral forebrain and posterior central nervous system continue to be expressed. This loss of GPC4 activity also causes apoptosis of forebrain progenitors during neural tube closure. Biochemical studies establish that GPC4 binds FGF2 and modulates FGF signal transduction. Inhibition of FGF signal transduction, by adding the chemical SU5402 to embryos from neural plate stages onwards, phenocopies the loss of gene expression and apoptosis in the forebrain. We propose that GPC4 regulates dorsoventral forebrain patterning by positive modulation of FGF signalling.


Studying the emergence of autism spectrum disorders in high-risk infants: methodological and practical issues.
Detecting early signs of autism is essential for timely diagnosis and initiation of effective interventions. Several research groups have initiated prospective studies of high-risk populations including infant siblings, to systematically collect data on early signs within a longitudinal design. Despite the potential advantages of prospective studies of young children at high-risk for autism, there are also significant methodological, ethical and practical challenges. This paper outlines several of these challenges, including those related to sampling (e.g., defining appropriate comparison groups), measurement and clinical implications (e.g., addressing the needs of infants suspected of having early signs). We suggest possible design and implementation strategies to address these various challenges, based on current research efforts in the field and previous studies involving high-risk populations.


[Results of electrical pacing in patients with hypertrophic obstruction cardiomyopathy (author's transl)].
In patients with hypertrophic obstructive cardiomyopathy, left ventricular outflow gradient decreased about 56 per cent by way of electrical stimulation of the apex of the right ventricle. In 4 patients a permanent pacemaker was implanted. P-wave triggered pacemakers with electronic conduction-time of approximately 0 sec showed to be preferable. The decrease of left ventricle outflow gradient as well as of subjective complaints inclusively cerebral syncopes were remarkable.


Body mass index and stroke risk among patients with diabetes mellitus in Korea.
Obesity and diabetes mellitus (DM) are both associated with cardiovascular disease. This study aimed to evaluate the association between body mass index (BMI) and stroke risk among patients with DM in Korea since relatively few studies have analyzed this area in detail. We analyzed a total of 56,051 DM patients aged >30 years from the Korean National Health Insurance Service Cohort who had undergone at least one national health examination between 2002 and 2012. BMI scores were divided into six categories, while hazard ratios for stroke were calculated using Cox proportional hazard models. Overall stroke risk was positively associated with BMI for both men and women. For ischemic stroke, the risk was positively associated with BMI in women. However, for me, only patients with the highest BMI were at increased risk compared with patients with a BMI of 20-22.4 kg/m2. For hemorrhagic stroke, the risk was significantly associated with BMI with a U-shaped association in men. In women, only patients with the lowest BMI had an increased risk of hemorrhagic stroke compared with patients that have a BMI of 20-22.4 kg/m2. BMI was positively associated with the overall risk of stroke among DM patients in Korea. The risk of ischemic stroke was higher in obese patients compared to overweight or normal-weight patients. However, the risk of hemorrhagic stroke was higher in slimmer patients compared with overweight or obese patients.


Peripheral type benzodiazepine receptor and airway smooth muscle relaxation.
It has been postulated that a benzodiazepine receptor with a micromolar affinity may be associated with Ca++ channels in peripheral organs. We examined the actions of Ro5-4684 (parachlorodiazepam) and midazolam on guinea pig tracheal smooth muscle contraction. Binding studies using [3H]Ro5-4684 indicate the presence of a "peripheral" type binding site with a Kd of approximately 4 nM and maximum binding of 1 pmol/mg of protein. Midazolam did not displace radioligand. In tension studies no activity was seen for Ro5-4684 or midazolam at concentrations below 1 microM. Higher concentrations relaxed the airway smooth muscle under basal tone, the effect was augmented significantly by epithelium removal. Similar results were obtained in tissues precontracted with methacholine or KCl. Midazolam (1 or 100 microM) significantly (P less than .05) attenuated the response to Ca++ in K+-depolarized tracheal strips, the effect was greater at low Ca++ concentrations. The compounds appear to function as Ca++ antagonists in airway smooth muscle but ar not typical as shown by their ability to reduce basal tone in airway smooth muscle.


Formula Feeding Predisposes Neonatal Piglets to Clostridium difficile Gut Infection.
Spontaneous outbreaks of Clostridium difficile infection (CDI) occur in neonatal piglets, but the predisposing factors are largely not known. To study the conditions for C. difficile colonization and CDI development, 48 neonatal piglets were moved into isolators, fed bovine milk-based formula, and infected with C. difficile 078. Analyses included clinical scoring; measurement of the fecal C. difficile burden, toxin B level, and calprotectin level; and postmortem histopathological analysis of colon specimens. Controls were noninfected suckling piglets. Fecal specimens from suckling piglets, formula-fed piglets, and formula-fed, C. difficile-infected piglets were used for metagenomics analysis. High background levels of C. difficile and toxin were detected in formula-fed piglets prior to infection, while suckling piglets carried about 3-fold less C. difficile, and toxin was not detected. Toxin level in C. difficile-challenged animals correlated positively with C. difficile and calprotectin levels. Postmortem signs of CDI were absent in suckling piglets, whereas mesocolonic edema and gas-filled distal small intestines and ceca, cellular damage, and reduced expression of claudins were associated with animals from the challenge trials. Microbiota in formula-fed piglets was enriched with Escherichia, Shigella, Streptococcus, Enterococcus, and Ruminococcus species. Formula-fed piglets were predisposed to C. difficile colonization earlier as compared to suckling piglets. Infection with a hypervirulent C. difficile ribotype did not aggravate the symptoms of infection. Sow-offspring association and consumption of porcine milk during early life may be crucial for the control of C. difficile expansion in piglets.


Anticancer effect of arsenic trioxide on cholangiocarcinoma: in vitro experiments and in vivo xenograft mouse model.
The purpose of our study was to investigate anticancer activity of arsenic trioxide (As2O3) on cholangiocarcinoma through in vitro and in vivo experiments using human cholangiocarcinoma cancer cells (CC-t6 cells) and a nude mouse model. The effect of As2O3 on CC-t6 cell survival was determined in vitro using MTT assay. Analysis of cell cycle phase distribution and quantification of apoptosis/necrosis, which were achieved by flow cytometry, were performed in order to understand the mechanism of As2O3. In vivo experiment was performed to assess the effectiveness of local injection of As2O3 on tumor inhibition by comparing the following three groups each consisting of five nude mouse xenograft models: high dose As2O3 (5 mg/kg), low dose As2O3 (1 mg/kg), and saline. In MTT assay, As2O3 inhibited the growth of CC-t6 cells more effectively than cisplatin or adriamycin at concentrations between 1 and 100 μM for most time points between 24 and 72 h (p < 0.05). With increased concentration of As2O3, there was dose-dependent increase in G 0/G 1 phase and dose-dependent decrease in S phase. As2O3-mediated inhibition of cell viability was achieved via induction of apoptosis and necrosis in a dose-dependent manner. Injection of As2O3 into CC-t6-induced tumors in nude mice inhibited the growth of subcutaneous tumor xenografts. As2O3 treatment dose-dependently inhibited the proliferation of CC-t6 cells via G 0/G 1 phase arrest and retarded tumor growth in nude mice, suggesting that As2O3 may be effective in the treatment of cholangiocarcinoma.


Galacto-Oligosaccharide RP-G28 Improves Multiple Clinical Outcomes in Lactose-Intolerant Patients.
Background and Aims: Lactose intolerance (LI) is a global problem affecting more than half of the world's population. An ultra-purified, high-concentration galacto-oligosaccharide, RP-G28, is being developed as a treatment for patients with LI. The efficacy and safety of RP-G28 in reducing symptoms of lactose intolerance were assessed in a blinded, randomized, placebo-controlled trial. Methods: In this multiclinical site, double-blinded, placebo-controlled trial, 377 patients with LI were randomized to one of two doses of orally administered RP-G28 or placebo for 30 days. A LI test and symptom assessment were performed at baseline and on day 31. The primary endpoint was a ≥4-point reduction or a score of zero on LI composite score on day 31. Voluntary milk and dairy intake and global outcome measures assessed patients' overall treatment satisfaction and quality of life before therapy and 30 days after therapy. This study received Institutional Review Board (IRB) approval. Results: For the primary endpoint, 40% in the RP-G28 groups reported a ≥4-point reduction or no symptoms on LI symptom composite score compared to 26% with placebo (P = 0.016). Treatment with RP-G28 also led to significantly higher levels of milk and dairy intake and significant improvements in global assessments compared to placebo. RP-G28 but not placebo led to significant increases in five Bifidobacterium taxa. Conclusions: RP-G28 for 30 days significantly reduced symptoms and altered the fecal microbiome in patients with LI. Treatment with RP-G28 also improved milk/dairy consumption and quality of life and was safe and well tolerated.


The mechanism of interleukin 4-induced down-regulation of CD38 on human B cells.
Exposure of Farage, a human B-cell line, to interleukin 4 (IL4) reduced the amount of CD38 antigen on the surface of the cells and in cell lysates. No evidence was obtained for accelerated breakdown, shedding, or internalization of CD38 molecules following IL4 treatment, nor the accumulation of CD38 molecules in the cell interior. The inhibition of protein synthesis with cycloheximide (CXM) diminished the down-regulation of CD38 induced by IL4. CXM decreased the expression of CD38 in Farage cells with arrested mitosis, and IL4 failed to further reduce CD38 expression. Staurosporine, an inhibitor of serine/threonine protein kinases, and H7 (1-(5-isoquinolinylsulfonyl)-2-methylpiperazine), a preferential inhibitor of protein kinase C (PKC), abrogated the effect of IL4 on CD38, while inhibitors of other serine protein kinases W7 (N-(aminohexyl)-5-chloro-1-naphthalenesulfoamide) and H8 (N-(2-[methylamino]ethyl)-5-isoquinolinesulfonamide) failed to interfere with the effect of IL4. Phorbol 12-myristate 13-acetate (PMA), an activator of PKC, resembled IL4 in decreasing the expression of CD38, and either staurosporine or H7 abolished this effect. Genistein, an inhibitor of tyrosine kinases, increased the expression of CD38, but failed to abrogate the inhibitory effect of IL4 on CD38. It is concluded that serine/threonine protein kinases mediated the IL4-induced down-regulation of the expression of CD38 molecules in B cells.


An Electrophysiological and Pharmacological Study of the Properties of Human iPSC-Derived Neurons for Drug Discovery.
Human stem cell-derived neurons are increasingly considered powerful models in drug discovery and disease modeling, despite limited characterization of their molecular properties. Here, we have conducted a detailed study of the properties of a commercial human induced Pluripotent Stem Cell (iPSC)-derived neuron line, iCell [GABA] neurons, maintained for up to 3 months in vitro. We confirmed that iCell neurons display neurite outgrowth within 24 h of plating and label for the pan-neuronal marker, βIII tubulin within the first week. Our multi-electrode array (MEA) recordings clearly showed neurons generated spontaneous, spike-like activity within 2 days of plating, which peaked at one week, and rapidly decreased over the second week to remain at low levels up to one month. Extracellularly recorded spikes were reversibly inhibited by tetrodotoxin. Patch-clamp experiments showed that iCell neurons generated spontaneous action potentials and expressed voltage-gated Na and K channels with membrane capacitances, resistances and membrane potentials that are consistent with native neurons. Our single neuron recordings revealed that reduced spiking observed in the MEA after the first week results from development of a dominant inhibitory tone from GABAergic neuron circuit maturation. GABA evoked concentration-dependent currents that were inhibited by the convulsants, bicuculline and picrotoxin, and potentiated by the positive allosteric modulators, diazepam, chlordiazepoxide, phenobarbital, allopregnanolone and mefenamic acid, consistent with native neuronal GABAA receptors. We also show that glycine evoked robust concentration-dependent currents that were inhibited by the neurotoxin, strychnine. Glutamate, AMPA, Kainate and NMDA each evoked concentration-dependent currents in iCell neurons that were blocked by their selective antagonists, consistent with the expression of ionotropic glutamate receptors. The NMDA currents required the presence of the co-agonist glycine and were blocked in a highly voltage-dependent manner by Mg2+ consistent with the properties of native neuronal NMDA receptors. Together, our data suggest that such human iPSC-derived neurons may have significant value in drug discovery and development and may eventually largely replace the need for animal tissues in human biomedical research.


Asymptomatic carotid bruit as related to single risk factors. A flowmetric study.
The prevalence of asymptomatic carotid bruit is 4 per cent in the general population aged over 40 years. According to most authors, it is an evident risk factor for stroke and myocardial infarction. In the present study the clinical data and Doppler flowmeter analysis of 5 groups of patients are reported. These patients had risk factors for cerebrovascular disease, without cerebral symptoms. An age matched group of healthy control subjects was also studied. The prevalence of haemodynamic stenosis (more than 50 per cent) of carotid artery is significantly higher among subjects with peripheral arteriopathy and arterial hypertension compared with other groups. On the other hand nonhaemodynamic stenosis (below 50 per cent) of carotid artery occurs to the same extent in patients and control groups. The possible implications of these findings on the knowledge of cerebrovascular obstructive disease are discussed.


Adult onset familial cherry-red spot myoclonus.
We report a case of a 36-year-old woman with progressive generalized myoclonus that first became apparent 9 years ago. Her younger brother had similar problems. Examination of her eyes revealed cherry-red spots. Hexosaminidase A, β-galactosidase and neuraminidase activity were normal. Although the laboratory findings were negative, cherry-red spots, progressive myoclonus and autosomal recessive inheritance pattern suggested that she had an unknown type of lysosomal storage disease.


Intra-role conflict: a result of naive attempts toward professionalization.
In an attempt to become professional, many allied health occupations have focused upon acquiring the traditional attributes associated with a profession while neglecting the professionalization process. As a result, members of allied health occupations that claim professional status may experience the problems of lack of occupational prestige, limited opportunity for vertical mobility, and occupational burn-out. A study was conducted to explore the possible links among level of training program socialization, level of professional identity, and level of intra-role conflict in medical technologists, registered dietitians, physicians' assistants, and certified dental technicians. An instrument was developed to test for the existence of relationships between intra-role conflict and a combination of factors. Results of this study support a relationship between occupation and training program socialization and intra-role conflict. Evidence in this study suggests that structural characteristics of an occupation may influence the level of intra-role conflict experienced by allied health practitioners.


Heat shock proteins; an overview.
Heat shock proteins (Hsps) protect protein substrates against conformational damage to promote the function of the proteins, prevent aggregation and prevent formation of toxic inclusion bodies. Protein aggregates and fibrils have been associated with neurodegenerative diseases and with inclusion bodies. High-level expression of recombinant protein for biotechnological purposes often leads to insoluble inclusion bodies. Therefore, misfolded proteins must be properly folded or must be degraded through heat shock protein action. This function protects cells against cytotoxic outcomes. In addition to their cytoprotective roles, Hsps are involved in other functions since Hsps exist in all types of cells and tissues. Therefore, several diseases are associated with alterations of these biochemical functions. This first review of the theme issue will discuss general properties of Hsps concisely along with their potential use in pharmaceutical and biotechnological applications.


Studies of activated GPIIb/IIIa receptors on the luminal surface of adherent platelets. Paradoxical loss of luminal receptors when platelets adhere to high density fibrinogen.
The accessibility of activated GPIIb/IIIa receptors on the luminal surface of platelets adherent to damaged blood vessels or atherosclerotic plaques is likely to play a crucial role in subsequent platelet recruitment. To define better the factors involved in this process, we developed a functional assay to assess the presence of activated, luminal GPIIb/IIIa receptors, based on their ability to bind erythrocytes containing a high density of covalently coupled RGD-containing peptides (thromboerythrocytes). Platelets readily adhered to wells coated with purified type I rat skin collagen and the adherent platelets bound a dense lawn of thromboerythrocytes. With fibrinogen-coated wells, platelet adhesion increased as the fibrinogen-coating concentration increased, reaching a plateau at about 11 micrograms/ml. Thromboerythrocyte binding to the platelets adherent to fibrinogen showed a paradoxical response, increasing at fibrinogen coating concentrations up to approximately 4-6 micrograms/ml and then dramatically decreasing at higher fibrinogen-coating concentrations. Scanning electron microscopy demonstrated that the morphology of platelets adherent to collagen was similar to that of platelets adherent to low density fibrinogen, with extensive filopodia formation and ruffling. In contrast, platelets adherent to high density fibrinogen showed a bland, flattened appearance. Immunogold staining of GPIIb/IIIa receptors demonstrated concentration of the receptors on the filopodia, and depletion of receptors on the flattened portion of the platelets. Thus, there is a paradoxical loss of accessible, activated GPIIb/IIIa receptors on the luminal surface of platelets adherent to high density fibrinogen. Two factors may contribute to this result: engagement of GPIIb/IIIa receptors with fibrinogen on the abluminal surface leading to the loss of luminal receptors, and loss of luminal filopodia that interact with thromboerythrocytes. These data provide insight into the differences in thrombogenicity between surfaces, and may provide a mechanism for purposefully passivating platelet-reactive artificial surfaces.


Voluntariness of consent to research: a conceptual model.
A good deal of policy and practice in human subjects research aims to ensure that when subjects consent to research, they do so voluntarily. To date, however, voluntariness and its impairment have been poorly conceptualized and studied. The legal doctrine of informed consent could provide a useful model.


Belgian atopic dermatitis guidelines.
Atopic dermatitis (AD) is one of the most common, bothersome and difficult to treat skin disorders. Recent introduction of new systemic treatments has revolutionized the management of AD. The goal of this guideline is to provide evidence-based recommendations for the management of patients suffering from atopic dermatitis that easily can be implemented in clinical practice. These recommendations were developed by 11 Belgian AD experts. Comments of all experts on the proposed statements were gathered, followed by an online voting session. The most relevant strategies for the management and treatment of AD in the context of the Belgian health care landscape are discussed. General measures, patient education and adequate topical treatment remain the cornerstones of AD management. For moderate to severe AD, the introduction of biologics and JAK inhibitors show unprecedented efficacy, although currently access is limited to a subgroup of patients meeting the reimbursement criteria.


Practice variations for fetal and neonatal congenital heart disease within the Children's Hospitals Neonatal Consortium.
Many aspects of care for fetuses and neonates with congenital heart disease (CHD) fall outside standard practice guidelines, leading to the potential for significant variation in clinical care for this vulnerable population. We conducted a cross-sectional survey of site sponsors of the Children's Hospitals Neonatal Consortium, a multicenter collaborative of 41 Level IV neonatal intensive care units to assess key areas of clinical practice variability for patients with fetal and neonatal CHD. We received responses from 31 centers. Fetal consult services are shared by neonatology and pediatric cardiology at 70% of centers. Three centers (10%) routinely perform fetal magnetic resonance imaging (MRI) for women with pregnancies complicated by fetal CHD. Genetic testing for CHD patients is routine at 76% of centers. Preoperative brain MRI is standard practice at 5 centers (17%), while cerebral NIRS monitoring is regularly used at 14 centers (48%). Use of electroencephalogram (EEG) after major cardiac surgery is routine in 5 centers (17%). Neurodevelopmental follow-up programs are offered at 30 centers (97%). Many aspects of fetal and neonatal CHD care are highly variable with evolving shared multidisciplinary models. Many aspects of fetal and neonatal CHD care are highly variable. Genetic testing, placental examination, preoperative neuroimaging, and postoperative EEG monitoring carry a high yield of finding abnormalities in patients with CHD and these tests may contribute to more precise prognostication and improve care. Evidence-based standards for prenatal and postnatal CHD care may decrease inter-center variability.


GenMiner: mining non-redundant association rules from integrated gene expression data and annotations.
GenMiner is an implementation of association rule discovery dedicated to the analysis of genomic data. It allows the analysis of datasets integrating multiple sources of biological data represented as both discrete values, such as gene annotations, and continuous values, such as gene expression measures. GenMiner implements the new NorDi (normal discretization) algorithm for normalizing and discretizing continuous values and takes advantage of the Close algorithm to efficiently generate minimal non-redundant association rules. Experiments show that execution time and memory usage of GenMiner are significantly smaller than those of the standard Apriori-based approach, as well as the number of extracted association rules. The GenMiner software and supplementary materials are available at http://bioinfo.unice.fr/publications/genminer_article/ and http://keia.i3s.unice.fr/?Implementations:GenMiner Supplementary data are available at Bioinformatics online.


Bayesian Robust Tensor Factorization for Incomplete Multiway Data.
We propose a generative model for robust tensor factorization in the presence of both missing data and outliers. The objective is to explicitly infer the underlying low-CANDECOMP/PARAFAC (CP)-rank tensor capturing the global information and a sparse tensor capturing the local information (also considered as outliers), thus providing the robust predictive distribution over missing entries. The low-CP-rank tensor is modeled by multilinear interactions between multiple latent factors on which the column sparsity is enforced by a hierarchical prior, while the sparse tensor is modeled by a hierarchical view of Student-t distribution that associates an individual hyperparameter with each element independently. For model learning, we develop an efficient variational inference under a fully Bayesian treatment, which can effectively prevent the overfitting problem and scales linearly with data size. In contrast to existing related works, our method can perform model selection automatically and implicitly without the need of tuning parameters. More specifically, it can discover the groundtruth of CP rank and automatically adapt the sparsity inducing priors to various types of outliers. In addition, the tradeoff between the low-rank approximation and the sparse representation can be optimized in the sense of maximum model evidence. The extensive experiments and comparisons with many state-of-the-art algorithms on both synthetic and real-world data sets demonstrate the superiorities of our method from several perspectives.


[Exposure to hand-arm vibrations in orthopaedic plaster room: risk management].
In hospitals, the use of vibrating tools, such as oscillating saws to cut plaster, can expose the staff to hand-arm vibrations. The aim of the study was to assess the exposure of workers to vibrations in the plaster room and then identify the most appropriate intervention for prevention and protection to be implemented in order to minimize exposure and protect workers' health, considering different individual hyper-susceptibility conditions. Four different models of plaster saws were examined for the evaluation. Various measurements were made in normal working conditions of the operators. The values of acceleration and noise detected on the instruments were in line with those reported in the literature. The preventive measure adopted (replacing plaster saws currently used in the hospital with similar ones with lower vibration emission) was an adequate means of protection. Health surveillance activities recorded a higher level of wellbeing, both environmentally and individually and, specifically, an increased protection level for the hyper-susceptibility conditions observed during health checks of exposed personnel which will be monitored regularly by the Occupational Health Service.


Maximal cytoreductive effort in epithelial ovarian cancer surgery.
The surgical management of advanced epithelial ovarian cancer involves cytoreduction, or removal of grossly-evident tumor. Residual disease after surgical cytoreduction of ovarian cancer has been shown to be strongly associated with survival. The goal of surgery is "optimal" surgical cytoreduction, which is generally defined as residual disease of 1 cm or less. However, the designation of "optimal" surgical cytoreduction has evolved to include maximal surgical effort and no gross residual disease. In order to achieve this, more aggressive surgical procedures such as rectosigmoidectomy, diaphragm peritonectomy, partial liver resection, and video-assisted thoracic surgery are reported and increasingly utilized in the surgical management of advanced ovarian cancer. The role of maximal surgical effort also extends to the recurrent setting where the goal of surgery should be complete cytoreduction. Patient selection is important in identifying appropriate candidates for surgical cytoreduction in the recurrent setting. The purpose of this article is to review the role of maximum surgical effort in primary and recurrent ovarian cancer.


Bias in tree searches and its consequences for measuring group supports.
When doing a bootstrap analysis with a single tree saved per pseudoreplicate, biased search algorithms may influence support values more than actual properties of the data set. Two methods commonly used for finding phylogenetic trees consist of randomizing the input order of species in multiple addition sequences followed by branch swapping, or using random trees as the starting point for branch swapping. The randomness inherent to such methods is assumed to eliminate any consistent preferences for some trees or unsupported groups of taxa, but both methods can be significantly biased. In the case of trees created by sequentially adding taxa, a bias may occur even if every addition sequence is equiprobable, and if one of the equally optimal positions for each terminal to add to the tree is selected equiprobably. In the case of branch swapping, the bias can happen even when branch swapping equiprobably selects any of the trees of better score in the subtree-pruning-regrafting-neighborhood or tree-bisection-reconnection-neighborhood. Consequently, when the data set is ambiguous, both random-addition sequences and branch swapping from random trees may (i) find some of the optimal trees much more frequently than others and (ii) find some groups with a frequency that differs from their frequency among all optimal trees. When the data set defines a single optimal tree, the groups present in that tree may have a different probability of being found by a search, even if supported by equal amounts of evidence. This may happen in both parsimony and maximum-likelihood analyses, and even in small data sets without incongruence.


[Fungal infections in oncohematology: the role of prophylaxis].
Fungal infections in patients with oncohematological diseases present a changing scene due to various circumstances such as: a better control of bacterial infections, the prophylactic use of antifungal agents such as fluconazole, the emergence of pathogens other than Candida spp., infections in non-neutropenic patients during graft-versus-host disease (GVHD), the use of other therapies that cause an increased immunosuppression (dexamethasone in high doses, fludarabine, rituximab, etc.), among others. Acute leukemia, hematopoietic transplant, GVHD, and treatment with purine analogs (fludarabine, cladribine) or alemtuzumab are considered to be the clinical scenarios with the highest risk. The ongoing management of fungal infections includes prophylaxis, empirical treatment of patients with fever not responsive to antibiotics, preemptive therapy, and specific treatment.


Improving mycetoma case detection through the training of community health workers in Northern Uganda: protocol for a stepped-wedge cluster-randomized trial.
The World Health Organization acknowledges the need for countries to incorporate neglected tropical disease care into their routine health care system. However, low detection rates and late presentation of mycetoma to health facilities have been observed in endemic countries, including Uganda. To empower community health workers (CHWs) in Northern Uganda to recognize and refer suspects of mycetoma to health facilities. This will be a stepped-wedge cluster-randomized trial based in Gulu and Pader districts over a period of 9 months with sequential crossover from intervention phase to the control phase at different time points until both districts are exposed to the intervention. The study will leverage on the ongoing partnership between Northern Uganda Medical Mission and the Uganda Ministry of Health that has trained over 300 CHWs in Gulu and Pader. The study evaluation will be done using the RE-AIM (Reach, Effectiveness, Adoption, Implementation, and Maintenance) framework. The expected outcome of the study is increased detection and referral of suspects of mycetoma. Data will be analyzed using STATA 17.0 and Friedman statistics or Analysis of Variance to determine increase in case identifications and referrals. The study was approved by Mulago Hospital Research and Ethics Committee (MHREC 2406) and registered with Pan African Clinical Trial Registry (PACTR202301534749787). The results from this trial will be published in a peer-reviewed journal. In addition, the findings will be shared at conferences, with funders, and at other research meetings.


Pseudomonas fluorescens HK44: lessons learned from a model whole-cell bioreporter with a broad application history.
Initially described in 1990, Pseudomonas fluorescens HK44 served as the first whole-cell bioreporter genetically endowed with a bioluminescent (luxCDABE) phenotype directly linked to a catabolic (naphthalene degradative) pathway. HK44 was the first genetically engineered microorganism to be released in the field to monitor bioremediation potential. Subsequent to that release, strain HK44 had been introduced into other solids (soils, sands), liquid (water, wastewater), and volatile environments. In these matrices, it has functioned as one of the best characterized chemically-responsive environmental bioreporters and as a model organism for understanding bacterial colonization and transport, cell immobilization strategies, and the kinetics of cellular bioluminescent emission. This review summarizes the characteristics of P. fluorescens HK44 and the extensive range of its applications with special focus on the monitoring of bioremediation processes and biosensing of environmental pollution.


Mechanism of hyperoxia-induced pulmonary vascular paralysis: effect of antioxidant pretreatment.
We designed experiments using isolated rabbit lungs to determine the effect of hyperoxia on the pulmonary vasoconstriction caused by the infusion of the lipid peroxide tert-butyl hydroperoxide (t-bu-OOH), which produces vasoconstriction by stimulating the pulmonary synthesis of thromboxane. Exposure to 48-60 h of 100% O2 at 1 ATA markedly reduced the increase in pulmonary artery pressure caused by t-bu-OOH infusion. We also investigated whether the mechanism for the attenuated vasoconstriction was due to altered production of arachidonate mediators or oxidant-induced damage to the contractile mechanism. In addition to infusing t-bu-OOH, which selectively stimulates thromboxane production, we also infused Intralipid, an esterified fatty acid emulsion that stimulates production of both thromboxane and prostacyclin. These experiments were done to study the effect of hyperoxia on prostacyclin synthesis. To determine if antioxidant therapy would prevent the changes in mediator production and vascular reactivity caused by hyperoxia, we pretreated animals with the antioxidants butylated hydroxyanisole (BHA) or vitamin E. The lack of vascular reactivity to t-bu-OOH was not due to a decrease in thromboxane synthesis or an increase in prostacyclin synthesis. Hyperoxia did not affect thromboxane synthesis during basal conditions or after stimulation of synthesis by t-bu-OOH. 100% O2 also did not effect the basal synthesis of prostacyclin by the lung. Hyperoxia did, however, markedly reduce prostacyclin synthesis when it was stimulated by Intralipid infusion. Antioxidant pretreatment did not reverse the inhibition of prostacyclin synthesis but did prevent the loss of vascular reactivity caused by hyperoxia. Thus hyperoxia causes vascular paralysis through oxidant-induced injury to the pulmonary vasculature.


An open-pollinated design for mapping imprinting genes in natural populations.
With the increasing recognition of its role in trait and disease development, it is crucial to account for genetic imprinting to illustrate the genetic architecture of complex traits. Genetic mapping can be innovated to test and estimate effects of genetic imprinting in a segregating population derived from experimental crosses. Here, we describe and assess a design for imprinting detection in natural plant populations. This design is to sample maternal plants at random from a natural population and collect open-pollinated (OP) seeds randomly from each maternal plant and germinate them into seedlings. A two-stage hierarchical platform is constructed to jointly analyze maternal and OP progeny markers. Through tracing the segregation and transmission of alleles from the parental to progeny generation, this platform allows parent-of-origin-dependent gene expression to be discerned, providing an avenue to estimate the effect of imprinting genes on a quantitative trait. The design is derived to estimate imprinting effects expressed at the haplotype level. Its usefulness and utilization were validated through computer simulation. This OP-based design provides a tool to detect the genomic distribution and pattern of imprinting genes as an important component of heritable variation that is neglected in traditional genetic studies of complex traits.


Semiconstrained total elbow arthroplasty for rheumatoid arthritis patients: clinical and radiological results of 1-8 years follow-up.
We investigated whether the Discovery total elbow arthroplasty (TEA) system had good results and survival in rheumatoid arthritis (RA) patients. In a prospective cohort study, one elbow surgeon performed TEA on 25 consecutive RA patients (31 elbows) between December 2004 and November 2012 using the Discovery system. We evaluated the preoperative elbow range of motion (ROM), functional outcome with QuickDash and quality of life with EQ-5D. An independent colleague evaluated the same parameters 1-8 years (mean 4.5) postoperatively. The medical records of the follow-up visits for the study period were available for review. A complete set of results was available for 19 patients (25 elbows). The mean ROM improved in flexion/extension from 88° (SD 27) to 113° (SD 19) and in pronation/supination from 55° (SD 28) to 68° (SD 22) (p < 0.05). The mean QuickDash also improved from 66.5 (SD 25.7) to 40.2 (SD 24) (p < 0.01). The mean EQ-5D improved from 0.68 (SD 0.2) to 0.75 (SD 0.13) but was not statistically significant (p = 0.09). Three patients were revised because of loosening, 2 more patients were re-operated. This resulted in a Kaplan-Meier survival of 90 % (CI 72-97) for the study period. The Discovery system has shown satisfactory results in RA patients even if the rate of complication remained relatively high. Further follow-up is required to investigate the radiological changes observed in some of our patients.


Quercetin and/or Ascorbic Acid Modulatory Effect on Phenobarbital-Induced Sleeping Mice Possibly through GABAA and GABAB Receptor Interaction Pathway.
Depressive disorder is a recurrent illness that affects large numbers of the general population worldwide. In recent years, the goal of depression treatment has moved from symptomatic response to that of full remission. However, treatment-resistant depression is a major challenge in the treatment of depression or depression-related disorders. Consensus opinion, therefore, suggests that effective combined aggressive initial treatment is the most appropriate strategy. This study aimed to evaluate the effects of quercetin (QUR) and/or ascorbic acid (AA) on Phenobarbital-induced sleeping mice. QUR (50 mg/kg) and/or AA (25 mg/kg) with or without intraperitoneally pre-treated with GABA receptor agonist (diazepam: 2 mg/kg, i.p.) or antagonist (Flumazenil: 2.5 mg/kg, i.p.) to underscore the effects, as well as the possible involvement of the GABA receptor in the modulatory action of QUR and AA in sleeping mice. Additionally, an in silico study was undertaken to predict the involvement of GABA receptors in the sleep mechanism. Findings suggest that the pretreatment of QUR and AA modulated the onset and duration of action of the standard drugs in experimental animals. The acute administration of QUR and/or AA significantly (p < 0.05) reversed the DZP-mediated onset of action and slightly reversed the duration of sleep time in comparison to the vehicle (control) group. A further combination of QUR or AA with the FLU resulted in an enhancement of the onset of action while reducing the duration of action, suggesting a FLU-like effect on the test animals. In in silico studies, AA and QUR showed good to moderate binding affinities with GABAA and GABAB receptors. Both QUR and AA produced a stimulatory-like effect on mice, possibly through the GABAA and GABAB receptor interaction pathways. Further studies are necessary to verify this activity and clarify the exact mechanism of action(s) involved.


Early neuroimaging and ultrastructural correlates of injury outcome after neonatal hypoxic-ischaemia.
Hypoxic ischaemia encephalopathy is the major cause of brain injury in new-borns. However, to date, useful biomarkers which may be used to early predict neurodevelopmental impairment for proper commencement of hypothermia therapy is still lacking. This study aimed to determine whether the early neuroimaging characteristics and ultrastructural correlates were associated with different injury progressions and brain damage severity outcomes after neonatal hypoxic ischaemia. Longitudinal 7 T MRI was performed within 6 h, 24 h and 7 days after hypoxic ischaemia in rat pups. The brain damage outcome at 7 days post-hypoxic ischaemia assessed using histopathology and MRI were classified as mild, moderate and severe. We found there was a spectrum of different brain damage severity outcomes after the same duration of hypoxic ischaemia. The severity of brain damage determined using MRI correlated well with that assessed by histopathology. Quantitative MRI characteristics denoting water diffusivity in the tissue showed significant differences in the apparent diffusion coefficient deficit volume and deficit ratios within 6 h, at 24 h and 7 days after hypoxic ischaemia among the 3 different outcome groups. The susceptible brain areas to hypoxic ischaemia were revealed by the temporal changes in regional apparent diffusion coefficient values among three outcome groups. Within 6 h post-hypoxic ischaemia, a larger apparent diffusion coefficient deficit volume and deficit ratios and lower apparent diffusion coefficient values were highly associated with adverse brain damage outcome. In the apparent diffusion coefficient deficit areas detected early after hypoxic ischaemia which were highly associated with severe damage outcome, transmission electron microscopy revealed fragmented nuclei; swollen rough endoplasmic reticulum and degenerating mitochondria in the cortex and prominent myelin loss and axon detraction in the white matter. Taken together, different apparent diffusion coefficient patterns obtained early after hypoxic ischaemia are highly associated with different injury progression leading to different brain damage severity outcomes, suggesting the apparent diffusion coefficient characteristics may be applicable to early identify the high-risk neonates for hypothermia therapy.


[Fasting-diet therapy of elderly patients with borderline mental disorders].
During the fasting dietetic therapy (FDT), 89 patients aged 46 to 75 years with mental disorders of nonpsychotic character (neurosis-like, neurotic and affective) were examined. The time-course of changes in the clinical status of patients with cerebral atherosclerosis, essential hypertension, slow-progredient schizophrenia, cyclothymia and manic-depressive psychosis, neuroses and lingering neurotic reactions during the FDT is described. The beneficial results in the form of considerable improvement and improvement of the mental status were attained in 83.2% of the patients. The elderly patients were found to tolerate FDT well. Side effects and somatic complications were recorded in 6 patients and were not serious. Based on the data obtained the FDT can be recommended for use on a wider basis in the management of elderly patients with borderline mental disorders.


Acute intermittent hypoxia induced neural plasticity in respiratory motor control.
Respiratory neural networks can adapt to rapid environmental change or be altered over the long term by various inputs. The mechanisms that underlie the plasticity necessary for adaptive changes in breathing remain unclear. Acute intermittent hypoxia (AIH)-induced respiratory long-term facilitation (LTF) is one of the most extensively studied types of respiratory plasticity. Acute intermittent hypoxia-induced LTF is present in several respiratory motor outputs, innervating both pump muscles (i.e. diaphragm) and valve muscles (i.e. tongue, pharynx and larynx). Long-term facilitation is present in various species, including humans, and the expression of LTF is influenced by gender, age and genetics. Serotonin plays a key role in initiating and modulating plasticity at the level of respiratory motor neurons. Recently, multiple intracellular pathways have been elucidated that are capable of giving rise to respiratory LTF. These mainly activate the metabolic receptors coupled to Gq ('Q' pathway) and Gs ('S' pathway) proteins. Herein, we discuss AIH-induced respiratory LTF in animals and humans, as well as recent advances in our understanding of the synaptic and intracellular pathways underlying this form of plasticity. We also discuss the potential to use intermittent hypoxia to induce functional recovery following cervical spinal injury.


Thirty-day outcomes of cerebrospinal fluid shunt surgery: data from the National Surgical Quality Improvement Program-Pediatrics.
Cerebrospinal fluid shunts are the mainstay of the treatment of hydrocephalus. In past studies, outcomes of shunt surgery have been analyzed based on follow-up of 1 year or longer. The goal of the current study is to characterize 30-day shunt outcomes, to identify clinical risk factors for shunt infection and failure, and to develop statistical models that might be used for risk stratification. Data for 2012 were obtained from the National Surgical Quality Improvement Program-Pediatrics (NSQIP-P) of the American College of Surgeons. Files with index surgical procedures for insertion or revision of a CSF shunt composed the study set. Returns to the operating room within 30 days for shunt infection and for shunt failure without infection were the study end points. Associations with a large number of potential clinical risk factors were analyzed on a univariate basis. Logistic regression was used for multivariate analysis. There were 1790 index surgical procedures analyzed. The overall rates of shunt infection and shunt failure without infection were 2.0% and 11.5%, respectively. Male sex, steroid use in the preceding 30 days, and nutritional support at the time of surgery were risk factors for shunt infection. Cardiac disease was a risk factor for shunt failure without infection, and initial shunt insertion, admission during the second quarter, and neuromuscular disease appeared to be protective. There was a weak association of increasing age with shunt failure without infection. Models based on these factors accounted for no more than 6% of observed variance. Construction of stable statistical models with internal validity for risk adjustment proved impossible. The precision of the NSQIP-P dataset has allowed identification of risk factors for shunt infection and for shunt failure without infection that have not been documented previously. Thirty-day shunt outcomes may be useful quality metrics, possibly even without risk adjustment. Whether important variation in 30-day outcomes exists among institutions or among neurosurgeons is yet unknown.


Renal denervation does not prevent hypertension in Lyon hypertensive rats.
The effects on blood pressure (BP) of a single and repeated renal denervations (RD) were compared in genetically hypertensive (LH) and normotensive (LN) rats of the Lyon strain. Indirect BP was measured by the tail-cuff method, and direct BP was measured through an aortic catheter in conscious quiet rats receiving a normal-sodium diet. Single RD (10 wk of age) did not reduce BP in both LN and LH rats. Repeated RD (4, 7, and 10 wk of age) partly prevented the age-related increase in tail-cuff BP but did not affect the directly recorded BP in 11-wk-old LH rats. Single and repeated RD reduced the renal norepinephrine content by greater than 90%. Both procedures did not alter the BP lowering effects of angiotensin-converting enzyme inhibition and of ganglionic blockade. It is concluded that renal nerves are not necessary for the development and the maintenance of hypertension in LH rats.


Combined light and electron microscopy using diaminobenzidine photooxidation to monitor trafficking of lipids derived from lipoprotein particles.
Diaminobenzidine (DAB) photooxidation is a method for conversion of fluorescent signals into electron-dense precipitates that are visible in the electron microscope. Recently, we have applied this method to analyze organelles involved in holo-high density lipoprotein (HDL) particle uptake at the ultrastructural level. In the present work we extended the spectrum of molecules visualized via photooxidation to monitor the uptake of HDL-derived lipids in HepG2 cells. By the combined light-electron microscopic method and with the aid of the DAB photooxidation technique, it became possible for the first time to visualize different intracellular pathways of lipoprotein particle-derived lipids and analyze the compartments involved at the ultrastructural level. HDL-Alexa 568 was used to visualize holo-HDL particle uptake. Reconstituted HDL particles containing the fluorescent cholesterol analogues Bodipy-cholesterol, Bodipy-cholesteryl oleate, or cholesteryl Bodipy-ester were used to visualize uptake of the HDL-associated sterol. In Bodipy-cholesteryl oleate and cholesteryl Bodipy-ester, the cholesterol moiety or the fatty acid moiety is fluorescently labeled, respectively; in contrast, Bodipy-cholesterol is an analogue of free cholesterol. The cellular compartments involved in their intracellular routes after uptake were analyzed in the fluorescence and electron microscope after DAB photooxidation. Bodipy-cholesterol was found to be localized in tubular endosomes and multivesicular bodies (MVBs), in the trans-Golgi network, and in stacked Golgi cisternae. In contrast, HepG2 cells incubated with HDL containing Bodipy-cholesteryl oleate or cholesteryl Bodipyester gave an uptake pattern comparable to that of holo-HDL particles, with MVBs being involved. Bodipy-cholesteryl oleate was also found in lysosomes. These results indicate that HDL-derived cholesterol and cholesteryl ester are transported by different intracellular pathways in HepG2 cells. Thus, the DAB photooxidation method enables the analysis of intracellular transport of lipoprotein particle-derived lipids at the light and at the ultrastructural level.


Visible spectrum waveguiding in bulk CMOS.
Waveguiding across the visible spectrum in an unmodified bulk CMOS chip is reported. The chip is fabricated in a standard CMOS process, and a simple wet etch removes metal in predetermined locations to expose glass rib waveguides. A modified Euler bend is introduced to improve bend radii by nearly an order of magnitude in the rib waveguides, and upper-bound losses are measured at visible wavelengths. These losses range from 6.2 dB/cm at 450 nm to 3.2 dB/cm at 650 nm and represent the lowest losses reported at visible wavelengths in unmodified bulk CMOS.


Neurons of the anterior mesial cortex related to faciovocal activity in the awake monkey.
1. The anterior mesial cortex, including the cingulate region, is thought to be involved in the voluntary control of vocalization. Previous recording studies have demonstrated that anterior mesial neurons discharge before conditioned and spontaneous vocalizations, but questions remain regarding the location and functional properties of these neurons. The present study was performed to provide a more complete description of the location and discharge properties of anterior mesial neurons involved in faciovocal behaviors. 2. Single-unit activity was recorded from neurons in the anterior mesial cortex of monkeys during performance of self-paced vocalizations and jaw openings. Cells were also tested for responsiveness to acoustic stimulation, and attempts were made to elicit vocalization through stimulation of the cortex surrounding related cells. Discharge properties of the cells were statistically analyzed, and correlation analysis was performed between measure of cell discharge and vocalization. 3. A total of 145 neurons were observed to modulate their discharge in association with vocalization or jaw opening. Four general classes of neurons were observed: neurons related only to vocalization, neurons related only to jaw opening, neurons related to both vocalization and jaw opening, and neurons related to other oromotor activities such as lip movements or reinforcement consumption. 4. Vocalization-related cells typically discharged tonically at a low frequency (mean 22 Hz), and many instances of long-lead activity (lead time > 500 ms) were noted. No neurons responded to acoustic stimulation, and electrical stimulation failed to elicit vocalization. Neural activity was not correlated with any measure of vocalization. 5. Neurons related to faciovocal behavior were located in the anterior cingulate sulcus and adjacent cortex of the mesial wall at a level just rostral to the genu of the arcuate sulcus. This region roughly corresponds to the rostral cingulate motor area and is located caudal to the traditionally described cingulate vocalization region. 6. In the present study we demonstrate the existence of an additional region in the medial wall that is involved in a variety of faciovocal behaviors such as vocalization, jaw opening, lip movements, and reinforcement consumption. The neurons do not appear to be strongly coupled to the execution of these acts. These results suggest that the activity of neurons in the anterior mesial cortex may relate to faciovocal behavior in a more global way than the activity of neurons in other motor areas.


High-Resolution Measurement of Local Activation Time Differences From Bipolar Electrogram Amplitude.
Localized changes in myocardial conduction velocity (CV) are pro-arrhythmic, but high-resolution mapping of local CV is not yet possible during clinical electrophysiology procedures. This is in part because measurement of local CV at small spatial scales (1 mm) requires accurate annotation of local activation time (LAT) differences with very high temporal resolution (≤1 ms), beyond that of standard clinical methods. We sought to develop a method for high-resolution measurement of LAT differences and validate against existing techniques. First, we use a simplified theoretical model to identify a quantitative relationship between the LAT difference of a pair of electrodes and the peak amplitude of the bipolar EGM measured between them. This allows LAT differences to be calculated from bipolar EGM peak amplitude, by a novel "Determination of EGM Latencies by Transformation of Amplitude" (DELTA) method. Next, we use simulated EGMs from a computational model to validate this method. With 1 kHz sampling, LAT differences less than 4 ms were more accurately measured with DELTA than by standard LAT annotation (mean error 3.8% vs. 22.9%). In a 1-dimensional and a 2-dimension model, CV calculations were more accurate using LAT differences found by the DELTA method than by standard LAT annotation (by unipolar dV/dt timing). DELTA-derived LAT differences were more accurate than standard LAT annotation in simulated complex fractionated EGMs from a model incorporating fibrosis. Finally, we validated the DELTA method in vivo using 18,740 bipolar EGMs recorded from the left atrium of 10 atrial fibrillation patients undergoing catheter ablation. Using clinical EGMs, there was agreement in LAT differences found by DELTA, standard LAT annotation, and unipolar waveform cross-correlation. These results demonstrate an underlying relationship between a bipolar EGM's peak amplitude and the activation time difference between its two electrodes. Our computational modeling and clinical results suggest this relationship can be leveraged clinically to improve measurement accuracy for small LAT differences, which may improve CV measurement at small spatial scales.