The endosomal lipid bis(monoacylglycero) phosphate as a potential key player in the mechanism of action of chloroquine against SARS-COV-2 and other enveloped viruses hijacking the endocytic pathway.
The anti-malarial drug Chloroquine (CQ) and its derivative hydroxychloroquine have shown antiviral activities in vitro against many viruses, including coronaviruses, dengue virus and the biosafety level 4 Nipah and Hendra paramyxoviruses. The in vivo efficacy of CQ in the treatment of COVID-19 is currently a matter of debate. CQ is a lysosomotrophic compound that accumulates in lysosomes, as well as in food vacuoles of Plasmodium falciparum. In the treatment of malaria, CQ impairs the digestion and growth of the parasite by increasing the pH of the food vacuole. Similarly, it is assumed that the antiviral effects of CQ results from the increase of lysosome pH and the inhibition of acidic proteases involved in the maturation of virus fusion protein. CQ has however other effects, among which phospholipidosis, characterized by the accumulation of multivesicular bodies within the cell. The increase in phospholipid species particularly concerns bis(monoacylglycero)phosphate (BMP), a specific lipid of late endosomes involved in vesicular trafficking and pH-dependent vesicle budding. It was shown previously that drugs like progesterone, the cationic amphiphile U18666A and the phospholipase inhibitor methyl arachidonyl fluoro phosphonate (MAFP) induce the accumulation of BMP in THP-1 cells and decrease cell infection by human immunodeficiency virus. HIV viral particles were found to be retained into large endosomal-type vesicles, preventing virus spreading. Since BMP was also reported to favour virus entry through hijacking of the endocytic pathway, we propose here that BMP could play a dual role in viral infection, with its antiviral effects triggered by lysosomotropic drugs like CQ.


Myostatin increases human trophoblast cell invasion by upregulating N-cadherin via SMAD2/3-SMAD4 signaling†.
Placental insufficiency disorders are major obstetric complications that share a common phenomenon of poor placental trophoblast cell invasion and remodeling of uterine tissues. Myostatin is a transforming growth factor (TGF)-β superfamily member well known for its important role in muscle growth control. Myostatin is also produced in the placenta and has been shown to regulate some trophoblast functions. However, its roles in placental development are still poorly understood. In this study, we tested the hypothesis that myostatin increases trophoblast cell invasion by upregulating N-cadherin via SMAD2/3-SMAD4 signaling. Primary and immortalized (HTR8/SVneo) trophoblast cells were used as study models. Matrigel-coated transwell invasion assays were used to study the effects of recombinant human myostatin on trophoblast cell invasion. Reverse transcription quantitative real-time polymerase chain reaction and Western blot were used to measure myostatin effects on N-cadherin mRNA and protein levels, respectively. Small inhibitor molecules as well as siRNA-mediated knockdown were used to block myostatin receptor and downstream signaling, respectively. Data were analyzed either by unpaired Student T test or one-way analysis of variance followed by Newman Keuls test for multiple group comparisons. Myostatin significantly increased primary and HTR8/SVneo trophoblast cell invasion. Moreover, myostatin upregulated N-cadherin mRNA and protein levels in a time-dependent manner in both study models. These effects were blocked by inhibition of TGF-β type I receptors as well as siRNA-mediated knockdown of SMAD2/3 combined or common SMAD4. Importantly, myostatin-induced trophoblast cell invasion was abolished by knockdown of N-cadherin, SMAD2/3, or SMAD4. Myostatin may increase human trophoblast cell invasion by upregulating N-cadherin via SMAD2/3-SMAD4 signaling.


Preliminary evaluations related to the ranges of hematological and biochemical variables in hospitalized patients with stroke.
According to the international reports, brain stroke is the main reason of death and disability. In ischemic stroke, early and precise classification of patients who may profit from conflicting finest therapeutic interference is necessary if enhanced effects in terms of survival are to be talented. Due to uncomplicated, easy performance, and inexpensive method the aim of this preliminary study was to investigate changes related to biochemical and hematological variables in patients with stroke. A cross-sectional study located at the neurology ward of the Ayatolah Kashani and Alzahra Hospitals' (conducted to Isfahan Neurosciences Research Center) was carried out on fifty patients (females; n = 20 and males; n = 30) between April 1, 2012 and September 31, 2012. The data from subjects' records were taken for analyzing variables. The statistical analysis of d-base was performed using (SPSS) for windows. Analysis of available data showed that with a mean of 182.4 mg/dl, blood sugar (BS) ranged from 75 to 300 mg/dl (n = 15/50). The changes in hemoglobin (Hgb) (mean 4.6 g/dl, n = 27/50), platelet (mean 210, 653/mm(3), n = 26/50) and lymphocyte (Lymph) (mean 37, n = 26/50) seems to be significant. The mean age of females was 76 years (ranged 46-93 years). The mean age of males was 70 years (ranged 31-90 years). Information related to previous drug history was available only in 24 patients. In 5 out of 22 cases ischemic heart disease (IHD) were positive. In 8 out of 29 cases, diabetes mellitus was positive. In 5 out of 28 cases, hypertension (HTN) was positive. In the four patients both IHD and HTN were positive. Any considerable alter in patients' biochemical and hematological figures (BS, Hgb, Plt and Lymph) may necessitate further attention related to inter- and intra-individual variability in clinical supervision and drug's assortment. Therefore, success in treatment could be achieved by the close management of clinical, biochemical, hematological, and pharmacological manifestation. To reduce disability, mortality, and morbidity in Iranian stroke population further clinical studies are needed to correlate drugs and laboratory markers to associated clinical events in order.


Therapeutic potential of autophagy-enhancing agents in Parkinson's disease.
Converging evidence from genetic, pathological and experimental studies have increasingly suggested an important role for autophagy impairment in Parkinson's Disease (PD). Genetic studies have identified mutations in genes encoding for components of the autophagy-lysosomal pathway (ALP), including glucosidase beta acid 1 (GBA1), that are associated with increased risk for developing PD. Observations in PD brain tissue suggest an aberrant regulation of autophagy associated with the aggregation of α-synuclein (α-syn). As autophagy is one of the main systems involved in the proteolytic degradation of α-syn, pharmacological enhancement of autophagy may be an attractive strategy to combat α-syn aggregation in PD. Here, we review the potential of autophagy enhancement as disease-modifying therapy in PD based on preclinical evidence. In particular, we provide an overview of the molecular regulation of autophagy and targets for pharmacological modulation within the ALP. In experimental models, beneficial effects on multiple pathological processes involved in PD, including α-syn aggregation, cell death, oxidative stress and mitochondrial dysfunction, have been demonstrated using the autophagy enhancers rapamycin and lithium. However, selectivity of these agents is limited, while upstream ALP signaling proteins are involved in many other pathways than autophagy. Broad stimulation of autophagy may therefore cause a wide spectrum of dose-dependent side-effects, suggesting that its clinical applicability is limited. However, recently developed agents selectively targeting core ALP components, including Transcription Factor EB (TFEB), lysosomes, GCase as well as chaperone-mediated autophagy regulators, exert more specific effects on molecular pathogenetic processes causing PD. To conclude, the targeted manipulation of downstream ALP components, rather than broad autophagy stimulation, may be an attractive strategy for the development of novel pharmacological therapies in PD. Further characterization of dysfunctional autophagy in different stages and molecular subtypes of PD in combination with the clinical translation of downstream autophagy regulation offers exciting new avenues for future drug development.


Giant axonal neuropathy in two siblings: clinical histopathological findings.
Giant axonal neuropathy is a rare, severe autosomal-recessive neurologic disease affecting both the peripheral and the central nervous system. In this article, we describe a detailed clinicopathological report of two affected sibs from a consanguineous Turkish family. The index patient was a 6.5-year-old girl. Her intellectual development was normal. At the age of 3, her parents noticed progressive lack of balance and deterioration of motor skills. On examination, she had paresis and sensory loss more marked distally. Her mental status was normal. Her older brother had similar findings. Electrophysiological studies of young patients showed decreased median and ulnar nerve conduction velocities, absent peroneal motor potential, absent sensory nerve potentials and an EMG suggesting a neurogenic pattern. MRI showed mild cerebral and cerebellar atrophy. The nerve biopsy showed moderate myelinated nerve fibres loss, several regenerative clusters and multiple giant axons. Focal demyelination, hypertrophic "onion pulp" changes and endoneural fibrosis were also seen. Immunohistochemically, neurofilament protein accumulation was detectable in giant axons. This consanguineous family with two affected siblings and healthy parents complies with autosomal-recessive inheritance in GAN. In the majority of reported GAN cases, CNS involvement is described early in the course of the disease, but these patients did not present any sign of CNS involvement. GAN is a rare genetic disease of childhood involving the central and peripheral nervous systems. The diagnosis is easy with clinical, electrophysiological, and histopathological features, if it has been done. Early diagnosis is important, because of possible prenatal diagnosis.


Human internal disc strains in axial compression measured noninvasively using magnetic resonance imaging.
Internal deformations and strains were measured within intact human motion segments. Quantify 2-dimensional internal deformation and strain in compression of human intervertebral discs using MRI. Experiments using radiographic or optical imaging have provided important data for internal disc deformations. However, these studies are limited by physical markers and/or disruption of the disc structural integrity. MR images were acquired before and during application of a 1000 N axial compression. Two-dimensional internal displacements, average strains, and the location and direction of peak strains were calculated using texture correlation, a pattern matching algorithm. The average height loss was 0.4 mm, which corresponded to 4.4% compressive strain. The inner AF radial displacement was outward, even with degeneration; the average outward displacement of the inner AF (0.16 mm) was less than the outer AF (0.36 mm). High shear peak strains (2%-26%) occurred near the endplate and at the inner AF. Shear was higher in the anterior AF compared to the posterior. This technique allows quantification of displacement and strain within the intact disc. The radial displacements of inner AF suggest NP translation under compression. Peak tensile radial strains occurred as vertical bands throughout the anulus, which may contribute to radial tears and herniations. The tensile axial and shear strains at the interface between the AF and endplate could be related to the occurrence of rim lesions. Peak strains at the endplate are likely due to the AF curvature and the oblique fibers angle at fiber insertion sites. In the future, this technique may be used to measure disc strain under a variety of loading conditions, such as bending or torsion, and could also be used to study the mechanical effects of disc degeneration and potential clinical interventions.


Maternal severity of illness across levels of care: a prospective, cross-sectional study.
The severity of illness of women experiencing severe maternal morbidity has not been quantified outside of the intensive care setting yet is likely to have a bearing on clinical needs. To examine severity of illness in women with severe maternal morbidity. A prospective observational study of critically ill pregnant and postpartum women was undertaken in intensive care units (ICU), high dependency units (HDU) and delivery suites (DS) of seven tertiary-level hospitals in Melbourne, during 2002-2004. Severity of illness was scored using the Acute Physiology and Chronic Health Evaluation version II (APACHE II) and Therapeutic Intervention Scoring System 28 items (TISS 28). 137 women participated in the study: ICU (n=33), HDU (n=46) and DS (n=58). The mean APACHE II score was 8.6 (95% CI 7.7-9.5) and mean TISS 28 score was 22.5 (95% CI 21.2-23.9). Women in ICU were sicker according to both APACHE II (mean 12.6, 95% CI 8.3-16.9) and TISS 28 (mean 31.5, 95% CI 28.2-35.5) compared to women not admitted to ICU (p<.005). There was no difference in the mean APACHE II scores of women in HDU (7.7, 95% CI 5.5-9.9) and DS (7.0, 95% CI 5.2-8.8; p=.20). Women born outside of Australia were more likely to be admitted to ICU (OR 3.27, 95% CI 1.19-8.97). Known risk factors like multiple pregnancy, age≥35 years and nulliparity were not associated with ICU admission. There was no difference in the severity of illness in women cared for in HDU and DS. It was not possible to predict which women would require ICU admission. Measurement of severity of illness adds a valuable dimension to the study of severe maternal morbidity.


The tramadol option.
Tramadol is an option for the treatment of rheumatological pain. Its mode of action and safety profile distinguishes it from other opioids. Tramadol differs from other opioids by combining a weak opioid and a monoaminergic mode of action. It is effective in different types of moderate-to-severe pain, including neuropathic pain. Moreover, as the mode of action of tramadol does not overlap with that of NSAIDs, it is a useful agent to be combined with these drugs. Tramadol induces fewer opioid adverse reactions for a given level of analgesia compared with traditional opioids. Common adverse reactions of tramadol such as nausea and dizziness, which usually occur only at the beginning of therapy and attenuate over time, can be further minimized by up-titrating the drug over several days. Dose adjustment is only necessary in patients over 75 years of age, or in those with either hepatic or renal insufficiency. Tramadol should be avoided or used with caution in epileptics, or in individuals who are receiving seizure-threshold lowering drugs. Finally, tramadol has a low risk of abuse because it has only a weak opioid effect and its monoaminergic action could inhibit the development of dependence. The low abuse potential of tramadol has been demonstrated by postmarketing surveillance data.


A Bayesian changepoint-threshold model to examine the effect of TMDL implementation on the flow-nitrogen concentration relationship in the Neuse River basin.
In-stream nutrient concentrations are well known to exhibit a strong relationship with river flow. The use of flow measurements to predict nutrient concentrations and subsequently nutrient loads is common in water quality modeling. Nevertheless, most adopted models assume that the relationship between flow and concentration is fixed across time as well as across different flow regimes. In this study, we developed a Bayesian changepoint-threshold model that relaxes these constraints and allows for the identification and quantification of any changes in the underlying flow-concentration relationship across time. The results from our study support the occurrence of a changepoint in time around the year 1999, which coincided with the period of implementing nitrogen control measures as part of the TMDL program developed for the Neuse Estuary in North Carolina. The occurrence of the changepoint challenges the underlying assumption of temporal invariance in the flow-concentrations relationship. The model results also point towards a transition in the river nitrogen delivery system from a point source dominated loading system towards a more complicated nonlinear system, where non-point source nutrient delivery plays a major role. Moreover, we use the developed model to assess the effectiveness of the nitrogen reduction measures in achieving a 30% drop in loading. The results indicate that while there is a strong evidence of a load reduction, there still remains a high level of uncertainty associated with the mean nitrogen load reduction. We show that the level of uncertainty around the estimated load reduction is not random but is flow related.


Femtosecond luminescence spectroscopy of core states in silicon nanocrystals.
We present a study of ultrafast carrier transfer from highly luminescent states inside the core of silicon nanocrystal (due to quasidirect transitions) to states on the nanocrystal-matrix interface. This transfer leads to a sub-picosecond luminescence decay, which is followed by a slower decay component induced by carrier relaxation to lower interface states. We investigate the luminescence dynamics for two different surface passivation types and we propose a general model describing spectral dependence of ultrafast carrier dynamics. Our results stress the crucial role of the energy distribution of the interface states on surface-related quenching of quasidirect luminescence in silicon nanocrystals. We discuss how to avoid this quenching in order to bring the attractive properties of the quasidirect recombination closer to exploitation.


Identification of proteins adsorbed from human plasma to glass bead columns: plasmin-induced degradation of adsorbed fibrinogen.
In a bead column experiment, attempts have been made to identify the proteins adsorbed from plasma onto a glass surface. Proteins adsorbed after a 3-h contact time were eluted sequentially by 1 M tris buffer and SDS. Polyacrylamide gel electrophoresis of eluted proteins showed a multiplicity of components, and not all of these could be identified. Positive identifications were made by immunodiffusion against specific antibodies, band positions on electrophoresis gels, and location of radioactivity in gels when specific radiolabeled proteins were added to plasma. Proteins found were albumin, IgG, fibrinogen, plasminogen, and fibrinogen degradation products (FDP). A major component with an apparent molecular weight of 25,000 remains unidentified. It is unrelated to albumin, IgG, fibrinogen, factor XII, or plasminogen. Adsorbed fibrinogen was less degraded when experiments were performed with plasmas deficient in either plasminogen or factor XII. It is therefore concluded that FDP are formed by activation of adsorbed plasminogen, as was found previously for purified fibrinogen containing a trace of plasminogen. At least part of this activation is potentiated by the contact activation phase of plasma coagulation, in particular activated factor XII.


Measuring quality of life in children with rhinoconjunctivitis.
The objective of this study was to develop, pretest, and validate a questionnaire to measure quality of life in children with seasonal allergic rhinoconjunctivitis (SAR). Thirty-four children with SAR were enrolled from summer camps, notices in the media, and an allergy clinic (Southern Ontario). After generating a pool of 48 potentially important quality of life items, the children identified the ones that they experienced with their SAR and scored each for bother (1 = a little bothered to 4 = extremely bothered). Items identified most frequently and with the highest bother score were included in the Paediatric Rhinoconjunctivitis Quality of Life Questionnaire (PRQLQ). The PRQLQ was pretested for ease completion and accuracy of understanding. The PRQLQ has 23 items in five domains (nose symptoms, eye symptoms, practical problems, other symptoms, and activities). Responses are given on a seven-point scale, and children are asked to score their experiences during the previous 7 days. Seventy-five children with symptomatic SAR were enrolled from notices in the media and a pediatric allergy clinic (Austin, Tex.). A single cohort design was used, with children assessed at 0, 1, and 3 weeks. The PRQLQ was administered to the children by a trained interviewer at 1 and 3 weeks. A conventional nasal symptom daily diary was completed for 1 week before each of these clinic visits. Global ratings were completed at the final visit. In patients who were stable between clinic visits, the PRQLQ demonstrated good reliability (intraclass correlation coefficient = 0.93). The questionnaire was very responsive to change (p < 0.001) and was able to differentiate between patients who were in a stable clinical state and those whose clinical state changed between visits (p = 0.005). Correlations between the PRQLQ and diary scores were close to predicted and supported both the cross-sectional and longitudinal validity of the PRQLQ. The PRQLQ measures the quality of life impairments important to children with SAR. Children provide reliable and accurate responses, the measurement properties are strong, and the questionnaire can be used with confidence in clinical trials, clinical practice, and surveys.


Alterations of acquired mouse mammary tumor virus DNA during mammary tumorigenesis in BALB/cfC3H mice.
The patterns of the milk-transmitted (exogenous) mouse mammary tumor virus (MuMTV) DNA restriction endonuclease fragments in the nodule and tumor stages of BALB/cfC3H mouse mammary neoplasia were compared with the use of the Southern blot analysis. Acquired MuMTV restriction fragments were detected in DNA from hyperplastic alveolar nodules (HAN), from primary hyperplastic outgrowths (HPO), from families of transplanted HPO, from tumors from HPO, and from serially transplanted tumors. The restriction fragment patterns suggested that the HAN were composed of clonal dominant populations. Transplantation of subdivisions of individual HAN resulted in HPO with DNA restriction patterns suggesting that HAN also contained two or more subpopulations. In all cases, HAN subpopulations shared MuMTV restriction fragments suggesting a common origin. Forty-seven tumors arising from HPO shared MuMTV restriction fragments with the HPO. Most but not all tumors had additional acquired MuMTV restriction fragments not detected in the progenitor HPO, indicating that they were composed of a distinct subpopulation that originated from the HPO. The restriction fragment pattern in some tumor lines was remarkably stable through many transplant generations. Some tumors had no major additional restriction fragments, suggesting that major rearrangements of MuMTV DNA are not required for tumorigenesis.


[Phenogenetic variability and population meronomy].
In accordance with M.A. Shishkin's epigenetic evolutionary theory and S.V. Meyen's concept of meron, N.P. Krenke's "phenogenetic variability" can be considered as a realization of developmentally determined laws of possible transformations of particular characters (morphogenetic realization of meron). It includes two components: deterministic (organizing) one and stochastic (random) one. Organizing includes (epigenetic variability) represents a canalized component of morphogenesis determined by creode structure and arrangement of epigenetic thresholds, which alows to speak about morphogenetic rule of meronomic transformations. Random (realized) variability corresponds with stochastic component of morphogenesis, which makes it possible a spontaneous shifting of available developmental programs and selection of alternative subcreodes. Concepts of "population epigenetic landscape", "population ontogenesis" and "population meronomy" are introduced. Population ontogenesis (PO) can be considered as a peculiar deformation of species developmental program common to all individuals in particular population. This deformation is historically adjusted to concrete environmental conditions by natural selection. PO reflects general set of potential developmental patterns in concrete population, and it should be peculiar and unique one in respect to the whole species. It may be assumed that each individual contains information about invariant population epigenetic landscape (PEL), and thereby discrete individual phenotypes represent a probabilistic copy of general population epigenetic pattern. Analysis of bilateral structures among members of the same generation permits to visualize the principal pattern of PEL. Epigenetic thresholds and constructional bans constrain probable morphogenetic transformations and creates PEL, which is a general rule-cliché that formats total disparity of character states and theirs stochastic manifestation in taxonomic units, populations and individuals. Based on Waddington's ideas of creode, S.V. Meyen introduced concepts of intracreode and extracreode. Population meronomy allows to characterize peculiarities of extracreode of a population or a taxonomic unit while studying intracreodes as intraindividual variability by means of phenetical analysis (reductional stage) and phenogenetic synthesis (compositional stage). In this case we estimate not individual variability (plasticity) proper but generalized population or taxonomic epigenetic diversity of intracreode, which in actually becomes an extracreode. Population-meronomic analysis of bilateral compositions of antimere structure elements allows to construct a natural system of structure transformations and thereby to visualize meronomic diversity and transformation paths of meron.


In vivo activation of latent HIV with a synthetic bryostatin analog effects both latent cell "kick" and "kill" in strategy for virus eradication.
The ability of HIV to establish a long-lived latent infection within resting CD4+ T cells leads to persistence and episodic resupply of the virus in patients treated with antiretroviral therapy (ART), thereby preventing eradication of the disease. Protein kinase C (PKC) modulators such as bryostatin 1 can activate these latently infected cells, potentially leading to their elimination by virus-mediated cytopathic effects, the host's immune response and/or therapeutic strategies targeting cells actively expressing virus. While research in this area has focused heavily on naturally-occurring PKC modulators, their study has been hampered by their limited and variable availability, and equally significantly by sub-optimal activity and in vivo tolerability. Here we show that a designed, synthetically-accessible analog of bryostatin 1 is better-tolerated in vivo when compared with the naturally-occurring product and potently induces HIV expression from latency in humanized BLT mice, a proven and important model for studying HIV persistence and pathogenesis in vivo. Importantly, this induction of virus expression causes some of the newly HIV-expressing cells to die. Thus, designed, synthetically-accessible, tunable, and efficacious bryostatin analogs can mediate both a "kick" and "kill" response in latently-infected cells and exhibit improved tolerability, therefore showing unique promise as clinical adjuvants for HIV eradication.


COMT: a common susceptibility gene in bipolar disorder and schizophrenia.
A variety of psychiatric illnesses, including schizophrenia and bipolar disorder, have been reported in patients with microdeletion on chromosome 22q11-a region which includes the catechol-O-methyltransferase (COMT) gene. The variety of psychiatric manifestations in patients with the 22q11 microdeletion and the role of COMT in the degradation of catecholamine neurotransmitters may thus suggest a general involvement of the COMT gene in psychiatric diseases. We have previously reported on a significant association between a COMT haplotype and schizophrenia. In this study, we attempt to test for association between bipolar disorder and the polymorphisms implicated in schizophrenia. The association between COMT and bipolar disorder was tested by examining the allele and haplotype found to be associated with schizophrenia. A significant association between bipolar disorder and COMT polymorphisms was found. The estimated relative risk is greater in women, a result consistent with our previous findings in schizophrenia. We suggest that polymorphisms in the COMT gene may influence susceptibility to both diseases--and probably also a wider range of behavioral traits.


The combination of DHEA, histamine, and insulin increases adipogenic differentiation and enhances tissue transplantation outcome in mice.
Adipose stem cells (ASCs) are pluripotent cells that can generate pure fat tissue for regeneration. Differentiated adipose cells have been generated by a common inducer cocktail composed of dexamethasone, insulin, and isobutylmethylxanthine (DIM). The major drawbacks of adipose cells are their tendency to float on the culture media and their cost. To overcome some of these disadvantages, a new inducer cocktail that includes insulin, dehydroepiandrosterone, and histamine (DH IH) was tested. As a result, lipid accumulation was elevated more than twofold with DH IH than with DIM. Cell adhesion and viability, which are important factors for stable differentiation, were increased with DH IH and were proven through measurement of mRNA expression levels of adhesion marker genes, N-cadherin and vascular cell adhesion molecule, as well as through an alamar blue assay. The expression of adipogenesis-related genes, adiponectin, and glucose transporter type 4 lasted for a long time. To improve the efficiency of grafting, cell adhesion and neovascularization need to be increased. Neovascularization was observed around the transplanted adipose cells, which showed a higher number of vessel formation in DH IH than in DIM. The above results suggest that DH IH can produce pure differentiated adipose cells effectively and enhance their adhesion onto the target location when these differentiated adipose cells were applied as a clinical resource.


From benign adrenal incidentaloma to adrenocortical carcinoma: an exceptional random event.
New European guidelines for the management of adrenal incidentalomas were recently released. One of the most novel recommendations is to stop following patients when they present a typical, small and non-secreting adenoma. We report here the case of a 71-year-old man with such an adenoma, who developed an adrenocortical carcinoma (ACC) fourteen years later, with subsequent metastases and death. Clinically, he had a normal blood pressure and no sign of hormonal hypersecretion. The hormonal work-up showed no hormone excess: urinary free cortisol level was normal, the diurnal cortisol rhythm was respected and urinary catecholamine metabolites levels were normal. Computed tomography (CT) scan showed a homogeneous lesion, with a low density. The lesion remained unchanged during the five years of follow-up. Eight years after the last CT, a large right heterogeneous adrenal mass was incidentally discovered during an ultrasound examination. On CT scan, it was a 6 cm heterogeneous tumor. On hormonal work-up, there was no secretion. The patient was operated of an adrenalectomy, and the histology described an ACC with a Weiss score at 8, with no benign contingent. To our knowledge, this is the first case of an ACC occurring in a patient with prior adrenal imaging showing a typical benign adenoma.


Relaxin increases the firing rate of supraoptic neurones and increases oxytocin secretion in the rat.
In urethane-anaesthetized ovariectomized rats, injection of porcine relaxin (7.5 and 15 micrograms/kg, i.v.) caused a sustained increase in circulating plasma oxytocin and vasopressin concentrations; 10 micrograms relaxin/rat i.v. produced a smaller but significant increase in plasma oxytocin concentration in conscious ovariectomized rats. A significant increase in oxytocin concentration and inhibition of the spontaneous milk-ejection reflex was also seen in anaesthetized (ovary intact) lactating rats following injection of relaxin (7.5 micrograms/kg, i.v.). To investigate whether relaxin acts by increasing the electrical activity of oxytocin neurones or by facilitating stimulus-secretion coupling in the pituitary, the electrical activity of neurones in the supraoptic nucleus was recorded in urethane-anaesthetized lactating rats and in ovariectomized rats. Porcine relaxin (10 micrograms/rat, i.v.) increased the firing rate of both oxytocin and vasopressin neurones in the supraoptic nucleus in lactating rats. The response to relaxin was unaffected by subsequent injection of naloxone (1 mg/kg, i.v.). Oxytocin neurones were also activated by injection of relaxin (10 micrograms/rat) into ovariectomized rats. Combining the electrophysiological data, the neuronal activation following relaxin was significantly correlated with the level of spontaneous activity prior to relaxin injection. The results show that relaxin acts centrally to increase circulating plasma oxytocin and vasopressin concentrations by an opioid-independent mechanism.


Size matters: gold nanoparticles in targeted cancer drug delivery.
Cancer is the current leading cause of death worldwide, responsible for approximately one quarter of all deaths in the USA and UK. Nanotechnologies provide tremendous opportunities for multimodal, site-specific drug delivery to these disease sites and Au nanoparticles further offer a particularly unique set of physical, chemical and photonic properties with which to do so. This review will highlight some recent advances, by our laboratory and others, in the use of Au nanoparticles for systemic drug delivery to these malignancies and will also provide insights into their rational design, synthesis, physiological properties and clinical/preclinical applications, as well as strategies and challenges toward the clinical implementation of these constructs moving forward.


Macrophages control the retention and trafficking of B lymphocytes in the splenic marginal zone.
The marginal zone of the spleen is a precisely ordered region that contains specialized subsets of B lymphocytes and macrophages. Disruption of the negative signaling inositol phosphatase, SH2-containing inositol-5-phosphatase 1 (SHIP), results in the loss of marginal zone B cells (MZBs) with reorganization of marginal zone macrophages (MZMOs) to the red pulp of the spleen. This primary macrophage defect, as revealed by selectively depleting SHIP in myeloid cells shows that MZMOs are specifically required for the retention of MZBs. The MZMO phenotype was reverted in SHIP/Bruton's tyrosine kinase (Btk) double knockout mice, thus identifying the Btk activating pathway as an essential component being regulated by SHIP. Furthermore, we identified a direct interaction between the MARCO scavenger receptor on MZMOs and MZBs. Activation or disruption of this interaction results in MZB migration to the follicle. The migration of the MZMOs was further studied after the response to Staphylococcus aureus, which induced MZMOs to move into the red pulp while MZBs migrated into the follicular zone. The marginal zone is therefore a dynamic structure in which retention and trafficking of B cells requires specific macrophage-B cell interactions.


Molecular detection by analysis of the 16S rRNA gene of fecal coliform bacteria from the two Korean Apodemus species (Apodemus agrarius and A. peninsulae).
Wild mouse feces can disseminate zoonotic microorganisms throughout a farm, which is a great threat to human health and can lead to economic loss through contaminated agricultural produce. To assess the microbial communities, especially fecal coliform bacteria, we used two methods. First, we isolated bacterial colonies onto the common media LB (lactose broth) agar, TSA (tryptic soy agar), and MRS (de Man, Rogosa, and Sharpe) agar, and then randomly select colonies from each plate and stocked them to the mother plate for genomic DNA isolation. Second, we analyzed bacterial colonies using the 16S rRNA gene molecular diagnostic method. Based on bacterial cultures and bacterial 16S rRNA gene markers, we detected four different bacterial species (Bacillus amyloliquefaciens, Escherichia coli, Staphylococcus xylosus, and Serratia liquefaciens) from fecal coliforms of the striped field mouse Apodemus agrarius and A. peninsulae in agricultural areas in South Korea. These results could help us to better understand the pathogen reservoirs of mice and initiate some preventive measures to mitigate the microbial risks associated with mouse fecal matter in agricultural production areas.


A new valid and reliable windtrainer VO2max protocol.
Maximal oxygen uptake (VO(2max)) cycling protocols usually require expensive laboratory ergometers where the athlete is unaccustomed to the cycling position. Recently, we developed a VO(2max) speed-ramped protocol with an indoor cycling simulator (Cateye Windtrainer WT) allowing cyclists to use their own bicycles during the VO(2max) test. The aim of this study was to test the validity of the WT protocol by comparing it with a traditional (TD) resistance-ramped protocol using an electronically braked ergometer. In addition, the retest reliability of the WT protocol was also determined. Twenty experienced cyclists randomly completed 6 VO(2max) protocols consisting of one familiarization trial and two tests on the WT and TD protocols. There were minimal differences in maximal oxygen uptake values between protocols (WT 64.1+/-7.1, TD 63.3+/-7.4 mL/kg/min). The variability in the difference of the means between the two protocols was 0.8 mL/kg/min (95% confidence interval CI: -0.26-2.02), the coefficient of variation (CV) was 2.8% (95%CI: 2.2-4.2%) and the interclass correlation was r=0.94 (P<0.01; 95%CI: 0.86-0.98). The intratest difference within the WT protocol was 1.5% (95%CI: -1.9-5%), CV=5.3% (95%CI: 4.1-8%) and the retest correlation was r=0.81 (P<0.01; 95%CI: 0.57-0.92). The WT speed-ramped protocol is a valid and reliable method to assess VO(2max).


[Culturally sensitive communication in end-of-life care: the care for Muslim patients as an example].
Suffering and death are an inevitable part of life. In our increasingly multicultural society, healthcare professionals are frequently confronted with ideas on suffering and death that are different from their own. As Muslims are the largest migrant group in the Netherlands, this article focuses specifically on their perspective, illustrated by a clinical case. The different experience of these phenomena, influenced by culture and religion, can lead to confusion and frustration for patients, their relatives, and healthcare professionals alike. It is essential that healthcare professionals are aware of both their own views and those of the patient, and have some knowledge of other cultures and religions. Healthcare professionals can use cultural (self-)reflection and culturally sensitive communication, examples of which are provided in this article, to build mutual trust and understanding. This may improve the patient-physician relationship and may make end-of-life communication, complex as it will ever be, a little more comprehensive.


Labor markets for health supply chain management in Rwanda: a qualitative study of stakeholder perspectives.
Effective supply chains for health products require an adequate, skilled workforce for supply chain management (SCM). Rwanda faces challenges in human resources for SCM, including limited capacity for SCM at different levels. Understanding of the factors influencing the supply of and demand for SCM professionals in Rwanda is necessary to ensure the labor market contains an adequate workforce. This study identifies the perspectives of key stakeholders in the supply chain management sector about the factors influencing the supply of and demand for SCM professionals. Data were collected in semi-structured group and one-on-one interviews with 39 key stakeholders involved in the supply chain management labor market between March and April 2019. Interviewees were categorized according to their role in the labor market as system actors, functional actors involved in the supply of SCM workers, and functional actors involved in the demand for SCM workers. Interviewees were asked open-ended questions about factors influencing the demand for and the supply of SCM workers, and recommendations for improvement. Interviews were analyzed thematically. One validation focus group was held and the results were also reviewed by the Ministry of Health in Rwanda. Stakeholders agreed that skills mismatch between SCM workers' skills and the competencies jobs require impacts the supply of workers. A lack of career structure for SCM, lack of professional definitions for health supply chain management, and SCM curricula that do not match the needs of the workplace contribute to this gap. The demand for SCM professionals is poorly defined in terms of the numbers of professionals needed and the skills workers require. Financial limitations hinder demand for health SCM professionals. This study adds to the understanding of factors influencing the SCM labor market in Rwanda by documenting perspectives from government ministries, professional organizations, universities, and employers from SCM organizations. Improving the SCM labor market in Rwanda and the availability of the skilled cadres required for the effective management of health supply chains in Rwanda requires a coordinated effort by the Ministry of Health in Rwanda, private SCM companies, professional associations, education sector, and policy makers.


Effectiveness of mHealth Interventions Targeting Health Care Workers to Improve Pregnancy Outcomes in Low- and Middle-Income Countries: A Systematic Review.
Low- and middle-income countries (LMICs) face the highest burden of maternal and neonatal deaths. Concurrently, they have the lowest number of physicians. Innovative methods such as the exchange of health-related information using mobile devices (mHealth) may support health care workers in the provision of antenatal, delivery, and postnatal care to improve maternal and neonatal outcomes in LMICs. We conducted a systematic review evaluating the effectiveness of mHealth interventions targeting health care workers to improve maternal and neonatal outcomes in LMIC. The Cochrane Library, PubMed, EMBASE, Global Health Library, and Popline were searched using predetermined search and indexing terms. Quality assessment was performed using an adapted Cochrane Risk of Bias Tool. A strength, weakness, opportunity, and threat analysis was performed for each included paper. A total of 19 studies were included for this systematic review, 10 intervention and 9 descriptive studies. mHealth interventions were used as communication, data collection, or educational tool by health care providers primarily at the community level in the provision of antenatal, delivery, and postnatal care. Interventions were used to track pregnant women to improve antenatal and delivery care, as well as facilitate referrals. None of the studies directly assessed the effect of mHealth on maternal and neonatal mortality. Challenges of mHealth interventions to assist health care workers consisted mainly of technical problems, such as mobile network coverage, internet access, electricity access, and maintenance of mobile phones. mHealth interventions targeting health care workers have the potential to improve maternal and neonatal health services in LMICs. However, there is a gap in the knowledge whether mHealth interventions directly affect maternal and neonatal outcomes and future research should employ experimental designs with relevant outcome measures to address this gap.


Efficacy of the Modifying Phonation Intervals (MPI) Stuttering Treatment Program With Adults Who Stutter.
This study compared a new adult stuttering treatment program (Modifying Phonation Intervals, or MPI) with the standard of care for reducing stuttered speech in adults (prolonged speech). Twenty-seven adults who stutter were assigned to either MPI or prolonged speech treatment, both of which used similar infrastructures. Speech and related variables were assessed in 3 within-clinic and 3 beyond-clinic speaking situations for participants who successfully completed all treatment phases. At transfer, maintenance, and follow-up, the speech of 14 participants who successfully completed treatment was similar to that of normally fluent adults. Successful participants also showed increased self-identification as a "normal speaker," decreased self-identification as a "stutterer," reduced short intervals of phonation, and some increased use of longer duration phonation intervals. Eleven successful participants received the MPI treatment, and 3 received the prolonged speech treatment. Outcomes for successful participants were very similar for the 2 treatments. The much larger proportion of successful participants in the MPI group, however, combined with the predictive value of specific changes in PI durations suggest that MPI treatment was relatively more effective at assisting clients to identify and change the specific speech behaviors that are associated with successful treatment of stuttered speech in adults.


Highly Cross-Linked Polyethylene Reduces Wear and Revision Rates in Total Hip Arthroplasty: A 10-Year Double-Blinded Randomized Controlled Trial.
Highly cross-linked polyethylene (XLPE) was developed to address the problem of wear and osteolysis associated with metal-on-conventional ultra-high molecular weight polyethylene (UHMWPE) bearing surfaces. The purpose of this study was to compare in vivo wear rates and clinical and radiographic outcomes between XLPE and UHMWPE in a prospective double-blinded randomized controlled trial with a minimum of 10 years of follow-up. We randomized 122 patients to receive either a conventional UHMWPE liner (Enduron; DePuy) or an XLPE liner (Marathon; DePuy). Ninety-one patients were assessed clinically and radiographically at a minimum of 10 years (range, 10.08 to 12.17 years). Oxford Hip Scores and Short Form-12 Health Survey scores were collected. The radiographs were analyzed for osteolysis and for 2-dimensional (2-D), 3-dimensional (3-D), and volumetric wear using validated software. All 122 patients were accounted for at the 10-year follow-up evaluation. Twelve patients had undergone revision surgery, 21 patients had died (1 of whom had previously undergone revision surgery), and 2 patients were unable to return for follow-up, leaving 91 patients available for clinical and radiographic evaluation. At a minimum of 10 years, 3-D wear rates were significantly lower (p < 0.001) in the XLPE group (mean, 0.03 mm/yr) than in the conventional UHMWPE group (mean, 0.27 mm/yr). The prevalence of osteolysis was also significantly lower in the XLPE group (38% versus 8%, p < 0.005), as was the revision rate (14.6% versus 1.9%, p = 0.012), with 10 of the 12 revisions being in the Enduron group. There was no significant difference between the clinical scores of the 2 groups. XLPE liners have significantly reduced wear and are associated with a greater implant survival rate at 10 years compared with conventional UHMWPE liners. Therapeutic Level I. See Instructions for Authors for a complete description of levels of evidence.


Effect of hyperbaric oxygen on experimental acute distal colitis.
It has been demonstrated that hyperbaric oxygen (HBO) is useful as an adjunctive therapy for Crohn's disease. However, its effects on ulcerative colitis have not been investigated. In the present study, HBO was tested for acetic acid-induced colitis, and antioxidant systems were evaluated to clarify its possible mode of action. Thirty-six Sprague-Dawley rats were randomly divided into three groups: sham control (Group I), colitis induced by acetic acid without any therapy (Group II), colitis induced by acetic acid and treated with HBO (Group III). HBO was given for 5 days, 2 sessions per day at 2.5-fold absolute atmosphere pressure (ATA) for a period of 90 min in rats in which colitis had been induced (Group III). Rats were sacrificed on the 5th day after the procedure. Superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH Px) activity were measured in the intestinal tissue and erythrocyte lysate. MDA and GSH Px were also determined in the plasma. Whereas MDA levels in erythrocyte, plasma and intestinal tissue were decreased, the levels of GSH Px and SOD were significantly increased in Group III as compared to those of Group II. The results of our study suggest that hyperbaric oxygen therapy has beneficial effects on the course of experimental distal colitis and that antioxidant systems may be involved in its mode of action.


Clinical and economic impact of flow cytometry crossmatching in primary cadaveric kidney and simultaneous pancreas-kidney transplant recipients.
We retrospectively compared the clinical and financial impact of a final cross-match by T cell flow cytometry (FXM) versus conventional complement-dependent cytotoxicity (CXM) in consecutive primary cadaveric kidney (K) and primary simultaneous cadaveric pancreas-kidney (SPK) transplant recipients. Mean follow-up was 14 months for both the K (range, 5-22 months) and SPK (range, 5-22 months) recipients. There were no instances of a positive CXM result if the FXM result was negative. However, 18 of the 102 (18%) K recipients and 11 of the 66 (17%) SPK recipients were FXM positive, CXM negative, but no grafts lost to hyperacute rejection in this group. In addition, patient survival, graft survival, incidence of acute rejection, and kidney and pancreas function (immediate and late) were not different in the FXM-positive versus the FXM-negative groups. Charges for the CXM and FXM methods were compared over a 6-month period. During that period, the FXM charges averaged $583 less per recipient than the CXM charges (58% reduction in charges), and the time required to perform the FXM method was 50% of that required for the CXM method. These results demonstrate that a clinical pathway for primary transplantation that utilizes the FXM rather than the CXM final cross-match is clinically safe, with no adverse effect on posttransplant outcome, reduces organ preservation time by shortening the waiting period for the final cross-match results, and significantly reduces the tissue typing charges. However, about 9% of all primary K and SPK recipients will be FXM positive, CXM negative on final cross-match and will be unnecessarily denied a transplant. In this study, we describe a method to identify these patients so that they can be tested by traditional CXM to avoid being denied access to donor organs.


Leukocyte trafficking: Can we bring the fight to the tumor?
Control of leukocyte trafficking plays a critical role in the establishment of effective immune responses. It is now well established that the number or ratio of effector to suppressor immune cells within the tumor microenvironment can significantly impact tumor growth and clinical outcomes. Recently approved immunotherapies by the FDA, and those in development, aim to stimulate effector immune cell function. For example, many checkpoint inhibitors seek to stimulate an immune response to tumors by reversing T-cell exhaustion. However, activation of the immune response outside the tumor microenvironment can lead to sometimes fatal immune-mediated adverse events -- the result of "on-target, off-tumor" effects. Thus, control of localization of these activated effector cells remains a critical component of optimizing tumor response while minimizing immune-mediated adverse events. Chemokines and chemoattractants, along with their receptors on immune cells, govern leukocyte trafficking; thus, understanding their expression pattern in the context of the tumor microenvironment and developing approaches to favorably alter those should lead to improved efficacy of current immunotherapeutics. This review highlights the background of cancer immunotherapy, leukocyte trafficking, and some novel approaches being utilized to optimize recruitment of effector immune cells into the tumor microenvironment. Future combinatorial immunotherapy should incorporate therapeutics aimed at 1) favorably altering the tumor microenvironment, 2) activating effector immune cells, and 3) optimizing effector cell trafficking into tumors.


Factors Associated with Carriage of Enteropathogenic and Non-Enteropathogenic Viruses: A Reanalysis of Matched Case-Control Data from the AFRIBIOTA Site in Antananarivo, Madagascar.
Environmental Enteric Dysfunction (EED) is an associate driver of stunting in poor settings, and intestinal infections indirectly contribute to the pathophysiology underlying EED. Our work aimed at assessing whether enteric viral carriage is determinant to stunting. A total of 464 healthy and asymptomatic children, aged 2 to 5 years, were recruited, and classified as non-stunted, moderately stunted, or severely stunted. Among the recruited children, 329 stool samples were obtained and screened for enteric and non-enteric viruses by real-time polymerase chain reaction. We statistically tested for the associations between enteric viral and potential risk factors. Approximately 51.7% of the stool samples were positive for at least one virus and 40.7% were positive for non-enteric adenoviruses. No statistical difference was observed between virus prevalence and the growth status of the children. We did not find any statistically significant association between viral infection and most of the socio-demographic risk factors studied, except for having an inadequate food quality score or an over-nourished mother. In addition, being positive for Ascaris lumbricoides was identified as a protective factor against viral infection. In conclusion, we did not find evidence of a direct link between stunting and enteropathogenic viral carriage in our population.


Clinical PhD graduate student views of their scientist-practitioner training.
The goal of the scientist-practitioner (S-P) training model is to produce clinical psychologists equipped to integrate and utilize both science and practice in the clinical and research domains. However, much has been written regarding the possible shortcomings of S-P training and whether clinical psychology graduate students are actually gaining the knowledge and skills to integrate science and practice during graduate training and beyond (Chang, Lee, & Hargreaves, 2008; Gelso, 2006; Merlo, Collins, & Bernstein, 2008; Phillips, 1993). As such, the present study assessed ratings of satisfaction, perception of ability, and use of the S-P training model within 653 clinical psychology graduate students enrolled in programs that are members of the Council of University Directors of Clinical Psychology. Findings suggest that students are consistently trained in the integration of science and practice and have confidence in their abilities to apply the S-P integration to research and clinical work. However, despite understanding the ways in which science can influence practice, over one third of students reported that they rarely use science-based decisions when informing clients of the clinical services they will be providing. The implications of these results support the need for a more detailed evaluation of clinical psychology graduate students as well as the use of research-informed practice and the process of providing clients with information they need to make informed choices about treatment.


Overweight status is associated with extensive signs of microvascular dysfunction and cardiovascular risk.
The aim of this present study was to investigate if overweight individuals exhibit signs of vascular dysfunction associated with a high risk for cardiovascular disease (CVD). One hundred lean and 100 overweight participants were recruited for the present study. Retinal microvascular function was assessed using the Dynamic Retinal Vessel Analyser (DVA), and systemic macrovascular function by means of flow-mediated dilation (FMD). Investigations also included body composition, carotid intimal-media thickness (c-IMT), ambulatory blood pressure monitoring (BP), fasting plasma glucose, triglycerides (TG), cholesterol levels (HDL-C and LDL-C), and plasma von Willebrand factor (vWF). Overweight individuals presented with higher right and left c-IMT (p = 0.005 and p = 0.002, respectively), average 24-h BP values (all p < 0.001), plasma glucose (p = 0.008), TG (p = 0.003), TG: HDL-C ratio (p = 0.010), and vWF levels (p = 0.004). Moreover, overweight individuals showed lower retinal arterial microvascular dilation (p = 0.039) and baseline-corrected flicker (bFR) responses (p = 0.022), as well as, prolonged dilation reaction time (RT, p = 0.047). These observations emphasise the importance of vascular screening and consideration of preventive interventions to decrease vascular risk in all individuals with adiposity above normal range.


Asymptomatic pulmonary embolism in lung cancer: prevalence and analysis of clinical and radiological characteristics in 141 outpatients.
Aims and background. The incidence of asymptomatic pulmonary embolism in cancer patients is unknown and strictly related to the imaging used for tumor assessment. Recent findings suggest a similar clinical outcome of asymptomatic pulmonary embolism events compared to symptomatic events with a significant impact on survival. The aim of the present study was to determine the prevalence of asymptomatic pulmonary embolism in a population of lung cancer outpatients and to investigate its clinical features. Methods. Outpatients with a diagnosis of lung carcinoma undergoing chemotherapy were selected from October 2006 to June 2009. Disease and patient characteristics, risk factors and treatment modalities were collected. All the computed tomography images performed for each patient during the study period were retrospectively reviewed to identify pulmonary embolism. Results. A total of 141 consecutive patients were included and 657 computed tomography scans were completely reviewed (from two to six consecutive scans for each patient). Asymptomatic pulmonary embolism in the study population had a prevalence of 14.9% (21 patients). Most of the events occurred in patients with adenocarcinoma, advanced stage and poor performance status, during the early phases of first-line chemotherapy or at the same time of the cancer diagnosis. Compared with the symptomatic pulmonary embolism events (5 patients), asymptomatic events occurred earlier (time from cancer diagnosis to pulmonary embolism of 3.5 [95% CI, 2.0-4.9] versus 12.1 months [95% CI, 6.3-17.9; P = 0.02]) and had a better prognosis (survival from PE of 7.5 [95% CI, 3.4-11.6] versus 1.9 months [95% CI, 0-3.9; P = 0.04]). Conclusions. Our findings indicate an underestimation of embolic events among lung cancer outpatients due to their frequent asymptomatic natur. Such a high prevalence suggests the importance to pay more attention to pulmonary embolism prevention in this population.


Differences in recruitment order of motor units in phasic and tonic flexion reflex in "spinal man".
The recruitment order of motoneurones in muscle contractions has been held to be largely constant and determined by the size of the cell. However, as shown in a previous investigation using electromyographic techniques, the order in which different motor units are activated during voluntary muscle contractions changes in normal human subjects on shifts from phasic to tonic contraction. In order to investigate these two types of activity also in cases in which the cerebral influence on the motoneurone pool is blocked, an analysis was made of the recruitment order in phasic and tonic flexion reflexes in 10 patients with total interruption of the spinal cord. The following four principles were found to apply and presumed to be generally valid for the isolated human spinal cord: (1) in the phasic exteroceptive reflex, the order of recruitment varies despite application of a standardized stimulus; (2) in the tonic reflex, the first unit to be recruited is usually the same even with widely different types of stimuli; (3) a shift from phasic to tonic reflex activation may result in considerable changes in recruitment order; (4) after facilitation by a subliminal long-lasting stimulus, the first unit to be recruited in the phasic reflex is also the first to be recruited in the tonic reflex. It is suggested that a tonic influence on the motoneurone pool is required for the presupposed constancy of the recruitment order.


Hypervariability of Nucleolus Organizer Regions in Bengal Slow Lorises, Nycticebus bengalensis (Primates, Lorisidae).
Slow lorises are a cryptic species complex, and thus genetic markers are needed to identify distinct evolutionary lineages or species. We examined the nucleolus organizer regions (NORs) of Bengal slow lorises (Nycticebus bengalensis) using FISH with 18S rDNA (rDNA-FISH) and silver nitrate staining (Ag-NOR stain). Ten individuals of the putatively single species N. bengalensis showed higher variability in localization than 3 other congeners, though their overall karyotypes were similar. The rDNA-FISH analysis detected a total of 18 loci, in contrast to previous studies of other slow loris species that revealed far fewer (6-10) loci. Eight of the 18 loci detected in the present analysis were found to be semi-stable localizations at 4 different chromosomes, while 10 were found to be unstable localizations at 5 other chromosomes. The semi-stable locations showed occasional presence/absence of variations for rDNA-FISH, and unstable locations were polymorphic among individuals, contributing to the higher variability of NORs in this taxon. We hypothesize that the larger numbers of rDNA loci found in N. bengalensis were introduced by genomic dispersion through ectopic recombination in association with terminal regions including rDNA. Such differences are potentially very powerful chromosomal markers to be used in species identification and conservation.


Characterization of two brassinosteroid C-6 oxidase genes in pea.
C-6 oxidation genes play a key role in the regulation of biologically active brassinosteroid (BR) levels in the plant. They control BR activation, which involves the C-6 oxidation of 6-deoxocastasterone (6-DeoxoCS) to castasterone (CS) and in some cases the further conversion of CS to brassinolide (BL). C-6 oxidation is controlled by the CYP85A family of cytochrome P450s, and to date, two CYP85As have been isolated in tomato (Solanum lycopersicum), two in Arabidopsis (Arabidopsis thaliana), one in rice (Oryza sativa), and one in grape (Vitis vinifera). We have now isolated two CYP85As (CYP85A1 and CYP85A6) from pea (Pisum sativum). However, unlike Arabidopsis and tomato, which both contain one BR C-6 oxidase that converts 6-DeoxoCS to CS and one BR C-6 Baeyer-Villiger oxidase that converts 6-DeoxoCS right through to BL, the two BR C-6 oxidases in pea both act principally to convert 6-DeoxoCS to CS. The isolation of these two BR C-6 oxidation genes in pea highlights the species-specific differences associated with C-6 oxidation. In addition, we have isolated a novel BR-deficient mutant, lke, which blocks the function of one of these two BR C-6 oxidases (CYP85A6). The lke mutant exhibits a phenotype intermediate between wild-type plants and previously characterized pea BR mutants (lk, lka, and lkb) and contains reduced levels of CS and increased levels of 6-DeoxoCS. To date, lke is the only mutant identified in pea that blocks the latter steps of BR biosynthesis and it will therefore provide an excellent tool to further examine the regulation of BR biosynthesis and the relative biological activities of CS and BL in pea.


Bronchial carcinoid appearing as a breast mass.
Carcinoid tumors of the breast are extremely rare. We studied what is to our knowledge the second case of an extramammary carcinoid presenting initially as a breast mass. Varying degrees of ribbon, trabecular, acinar, and insular patterns were present in the primary bronchial tumor and in several secondary deposits, including the initial breast lesion. The cells were argyrophilic and contained round and membrane-bound electron-dense granules. Estrogen and progesterone receptors were present in a cerebral metastasis. Polypeptide hormones and 5-hydroxytryptamine were not detected by immunocytochemistry.


Ginsenoside compound-K inhibits the activity of B cells through inducing IgD-B cell receptor endocytosis in mice with collagen-induced arthritis.
Previously, ginsenoside metabolite compound K (C-K) was able to reduce B cell proliferation and serum anti-type II collagen (anti-CII) antibody to normal levels in mice with collagen-induced arthritis (CIA); however, the mechanism by which C-K restores B cell balance is unclear. In the present work, C-K treatment not only alleviated the polyarthritis index, swollen joint count, pathological scores of spleen and joints, spleen index, B cell proliferation and the level of serum antibodies (IgG1, IgG2a and anti-collagen II), but C-K treatment also restored B cell subsets including regulatory B cells, plasma cells, memory B cells, mature B cells, and follicular B cells in CIA mice. Interestingly, C-K did not change the expression level of immunoglobulin D-type B-cell receptor (IgD-BCR) but promoted IgD-BCR endocytosis. C-K treatment enhanced β-arrestin1 expression, facilitating the colocalization between IgD and β-arrestin1, as well as colocalization between IgD and adaptor protein 2 (AP2). Inhibition of the β-arrestin1-AP2 interaction with barbadin significantly reduced the ability of C-K to attenuate IgD-BCR plasma membrane localization. These results taken together depict that C-K ameliorates CIA in part by inhibiting B cell activation through the triggering of IgD-BCR internalization in a β-arrestin1-AP2 dependent manner.


Increased fixation of sulfur-35 by cartilage in vitro following depletion of the matrix by intravenous papain.
The uptake in vitro of sulfur-35 by costal cartilage obtained from nine rabbits 11 days after an intravenous injection of crude papain solution was compared with that in costal cartilage from eight normal untreated rabbits. An increased fixation of the isotope was found in treated animals compared with controls. The depletion of cartilage matrix by papain provided an experimental situation to test the hypothesis that the depletion of matrix which occurs in osteoarthritic cartilage can stimulate increased synthesis of chondroitin sulfate. The results give further support to the view that the primary lesion in osteoarthritis occurs in the matrix rather than in the chondrocyte of articular cartilage.


THE CHLOROPLAST H+-ATPase: PARTIAL REACTIONS OF THE PROTON.
This article reviews proton intake, charge transfer and proton release by F-ATPases, based in part on flash spectrophotometric studies on the chloroplast ATPase in thylakoid membranes, CF1Fo. The synthesis-coupled translocation of charges by CF1Fo (maximum rate <1500 s-1) and the dissipative flow through its exposed channel portion, CFo (rate >10 000 s-1), are extremely proton-specific (selectivity H+:K+>10(7):1). The proton-specific filter is located in CFo. Proton flow through exposed CFo can be throttled by adding subunit (&dgr;) or subunit &bgr; of CF1. These subunits thus may provide energy-transducing contacts between CF1 and CFo. Recently, we characterized two conditions where, in contrast to the above situation, proton intake by CF1Fo was decoupled from proton transfer across the main dielectric barrier: (a) CF1Fo structurally distorted by low ionic strength transiently trapped protons in a highly cooperative manner, but remained proton tight. This result has been interpreted in terms of Mitchell's proton well. (b) In the absence of nucleotides there is a proton slip. Addition of nucleotides (100 nmol l-1 ADP) abolished proton conduction but not proton intake by CF1Fo. These experiments functionally tag proton binding groups on CF1Fo that are located before the main dielectric barrier.


Hydrogen gas production of broiler chicks in response to soybean meal and alpha-galactoside free, ethanol-extracted soybean meal.
In order to measure broiler chick hydrogen gas production, a sealed atmosphere chamber was constructed and chicks were intubated with soybean meal (SBM), alpha-galactoside free, ethanol-extracted soybean meal (ESBM), and ESBM with alpha-galactosides added to the levels of SBM (ESBMG). Six male broiler chicks averaging 156 g of weight were deprived of feed for 12 h prior to intubation with 6 g of the test soybean meals. Two chicks were used for each treatment. Following intubation, chicks were placed in the sealed atmosphere chamber for 20 min at 2-h intervals for 28 h. At the end of this 20-min period, a sample of the chamber atmosphere was collected with a gas-tight syringe and analyzed for hydrogen gas by gas-solid chromatography. The hydrogen production of the two chicks intubated with SBM peaked 7 h postintubation at 127 ppm. The ESBM produced a peak at approximately 17 h postintubation at 26 ppm. Intubation with ESBMG resulted in peak hydrogen production at approximately 12 h postintubation at an average of 67 ppm. Results indicate that chicks intubated with SBM produced 3.2 times the amount of total hydrogen gas than those chicks intubated with ESBM. Chicks intubated with ESBMG produced 2.2 times the amount of total hydrogen gas than chicks intubated with ESBM. The research indicates the alpha-galactoside oligosaccharides are a major cause of hydrogen gas production from SBM in poultry.


Associations between the gut microbiome and metabolome in early life.
The infant intestinal microbiome plays an important role in metabolism and immune development with impacts on lifelong health. The linkage between the taxonomic composition of the microbiome and its metabolic phenotype is undefined and complicated by redundancies in the taxon-function relationship within microbial communities. To inform a more mechanistic understanding of the relationship between the microbiome and health, we performed an integrative statistical and machine learning-based analysis of microbe taxonomic structure and metabolic function in order to characterize the taxa-function relationship in early life. Stool samples collected from infants enrolled in the New Hampshire Birth Cohort Study (NHBCS) at approximately 6-weeks (n = 158) and 12-months (n = 282) of age were profiled using targeted and untargeted nuclear magnetic resonance (NMR) spectroscopy as well as DNA sequencing of the V4-V5 hypervariable region from the bacterial 16S rRNA gene. There was significant inter-omic concordance based on Procrustes analysis (6 weeks: p = 0.056; 12 months: p = 0.001), however this association was no longer significant when accounting for phylogenetic relationships using generalized UniFrac distance metric (6 weeks: p = 0.376; 12 months: p = 0.069). Sparse canonical correlation analysis showed significant correlation, as well as identifying sets of microbe/metabolites driving microbiome-metabolome relatedness. Performance of machine learning models varied across different metabolites, with support vector machines (radial basis function kernel) being the consistently top ranked model. However, predictive R2 values demonstrated poor predictive performance across all models assessed (avg: - 5.06% -- 6 weeks; - 3.7% -- 12 months). Conversely, the Spearman correlation metric was higher (avg: 0.344-6 weeks; 0.265-12 months). This demonstrated that taxonomic relative abundance was not predictive of metabolite concentrations. Our results suggest a degree of overall association between taxonomic profiles and metabolite concentrations. However, lack of predictive capacity for stool metabolic signatures reflects, in part, the possible role of functional redundancy in defining the taxa-function relationship in early life as well as the bidirectional nature of the microbiome-metabolome association. Our results provide evidence in favor of a multi-omic approach for microbiome studies, especially those focused on health outcomes.


Granulocyte-associated IgG in neutropenic disorders.
We applied a radiolabeled antiglobulin test to a study of patients with a variety of neutropenic disorders. After defining the nature of the interaction of radiolabeled anti-IgG with the neutrophil, we studied 16 patients with neutropenia of uncertain etiology and adequate bone marrow granulocyte precursors. Twelve of these 16 patients had increased neutrophil-associated IgG (PMN-IgG). Patients with the highest levels of PMN-IgG had the lowest neutrophil counts. The majority of patients with neutropenia and increased PMN-IgG had an underlying immunologic disorder that included immune thrombocytopenic purpura in 5 patients and autoimmune hemolytic anemia in 1 patient. In some patients, elevated PMN-IgG preceded other evidence for immunologic disease. The direct antiglobulin test helped to distinguish neutropenic patients with increased PMN-IgG both from patients with neutropenia due to a known nonimmune disorder and from nonneutropenic patients with rheumatoid arthritis or systemic lupus erythematosis. Each of four patients with increased neutrophil-associated IgG treated with systemic corticosteroids responded clinically with an associated fall in neutrophil IgG and a rise in the circulating neutrophil count. The radiolabeled antiglobulin test appears useful in defining a subpopulation of patients with neutropenia due to an underlying immunologic disorder.


Dielectric nanosheet modified plasmonic-paper as highly sensitive and stable SERS substrate and its application for pesticides detection.
The interaction of plasmonic nanoparticles with a dielectric platform gives rise to unique optical behaviors and this can be maneuvered to improve the plasmonic/SERS performances of a substrate. Herein, dielectric modified plasmonic-paper SERS substrate is developed by assembling Ag@SiO2 nanocubes on Fe-TiO2 nanosheets (NS) modified paper. The Fe-TiO2 NS being visible light responsive significantly alters the optical property of the paper and serves as a dielectric underlay for the Ag nanocubes. Hence, the incident light reflected back from the dielectric nanosheets couples with the scattered light from the Ag nanocubes leading to spatially enhanced electromagnetic field improving the SERS enhancement. The prepared dielectric modified plasmonic-paper has an average enhancement factor (EF) of 1.49 × 107 using R6G as a probe molecule. This value is superior to unmodified plasmonic-paper highlighting the coupling effect of the dielectric nanosheets. The substrate shows robust detection performance for thiabendazole and achieves a limit of detection (LOD) of 19 μg/L, which is 4-fold more sensitive than unmodified plasmonic paper. Direct swabbing test of thiabendazole sprayed apple fruit shows a discernible Raman signal down to 15 ppb indicating the utility of the substrate for point-of-need applications in food safety.


Management of inadvertent intraocular injections.
A case of inadvertent intraocular injection of lidocaine is described. The needle entered the eye through the pars plana, without causing any trauma to the anterior segment or retina. The pharmacologic effects of intraocular lidocaine could be determined in this case, which had no complications attributable to concurrent physical damage to the eye. Following the injection, the visual acuity was no light perception; vision ultimately returned to 20/20. Appropriate medical and surgical management of such cases is discussed.


Central endogenous angiotensin-(1-7) protects against aldosterone/NaCl-induced hypertension in female rats.
In comparison to male rodents, females are protected against angiotensin (ANG) II- and aldosterone (Aldo)-induced hypertension. However, the mechanisms underlying this protective effect are not well understood. ANG-(1-7) is formed from ANG II by angiotensin-converting enzyme 2 (ACE2) and has an antihypertensive effect in the central nervous system (CNS). The present study tested the hypothesis that central ANG-(1-7) plays an important protective role in attenuating the development of Aldo/NaCl-hypertension in female rats. Systemic infusion of Aldo into intact female rats with 1% NaCl as their sole drinking fluid resulted in a slight increase in blood pressure (BP). Intracerebroventricular (icv) infusion of A-779, an ANG-(1-7) receptor (Mas-R) antagonist, significantly augmented the pressor effects of Aldo/NaCl. In contrast, systemic Aldo/NaCl induced a significant increase in BP in ovariectomized (OVX) female rats, and central infusion of ANG-(1-7) significantly attenuated this Aldo/NaCl pressor effect. The inhibitory effect of ANG-(1-7) on the Aldo/NaCl pressor effect was abolished by concurrent infusion of A-779. RT-PCR analyses showed that there was a corresponding change in mRNA expression of several renin-angiotensin system components, estrogen receptors and an NADPH oxidase subunit in the lamina terminalis. Taken together these results suggest that female sex hormones regulate an antihypertensive axis of the brain renin-angiotensin system involving ACE2/ANG-(1-7)/Mas-R that plays an important counterregulatory role in protecting against the development of Aldo/NaCl-induced hypertension.


The ubiquitin-proteasome pathway and its role in cancer.
Critical cellular processes are regulated, in part, by maintaining the appropriate intracellular levels of proteins. Whereas de novo protein synthesis is a comparatively slow process, proteins are rapidly degraded at a rate compatible with the control of cell cycle transitions and cell death induction. A major pathway for protein degradation is initiated by the addition of multiple 76-amino acid ubiquitin monomers via a three-step process of ubiquitin activation and substrate recognition. Polyubiquitination targets proteins for recognition and processing by the 26S proteasome, a cylindrical organelle that recognizes ubiquitinated proteins, degrades the proteins, and recycles ubiquitin. The critical roles played by ubiquitin-mediated protein turnover in cell cycle regulation makes this process a target for oncogenic mutations. Oncogenes of several common malignancies, for example colon and renal cell cancer, code for ubiquitin ligase components. Cervical oncogenesis by human papillomavirus is also mediated by alteration of ubiquitin ligase pathways. Protein degradation pathways are also targets for cancer therapy, as shown by the successful introduction of bortezomib, an inhibitor of the 26S proteasome. Further work in this area holds great promise toward our understanding and treatment of a wide range of cancers.


Comparison of static-fluid or excretory magnetic resonance urography with computed tomography urography for visualization of nondilated renal pelvises and ureters in healthy Beagles.
To assess the usefulness of magnetic resonance urography (MRU) for the visualization of nondilated renal pelvises and ureters in dogs and to compare our findings for MRU versus CT urography (CTU). 9 healthy Beagles. Dogs underwent CTU, static-fluid MRU, and excretory MRU, with ≥ 7 days between procedures. Contrast medium was administered IV during CTU and excretory MRU, whereas urine in the urinary tract was an intrinsic contrast medium for static-fluid MRU. For each procedure, furosemide (1 mg/kg, IV) was administered, and reconstructed dorsal plane images were acquired 3 minutes (n = 2) and 7 minutes (2) later. Images were scored for visualization of those structures and for image quality, diameters of renal pelvises and ureters were measured, and results were compared across imaging techniques. Excretory MRU and CTU allowed good visualization of the renal pelvises and ureters, whereas static-fluid MRU provided lower visualization of the ureters. Distention of the renal pelvises and ureters was good in excretory MRU and CTU. Distention of the ureters in static-fluid MRU was insufficient compared with that in CTU and excretory MRU. Distinct artifacts were not observed in CTU and excretory MRU images. Static-fluid MRU images had several mild motion artifacts. Our findings indicated that excretory MRU with furosemide administration was useful for visualizing nondilated renal pelvises and ureters of dogs in the present study. When performing MRU for the evaluation of dogs without urinary tract dilation, excretory MRU may be more suitable than static-fluid MRU.


Total Synthesis and Complete Stereostructure of a Marine Macrolide Glycoside, (-)-Lyngbyaloside B.
We have described in detail the total synthesis of both the proposed and correct structures of (-)-lyngbyaloside B, which facilitated the elucidation of the complete stereostructure of this natural product. Our study began with the total synthesis of 13-demethyllyngbyaloside B, in which an esterification/ring-closing metathesis (RCM) strategy was successfully used for the efficient construction of the macrocycle. We also established reliable methods for the introduction of the conjugated diene side chain and the l-rhamnose residue onto the macrocyclic framework. However, the esterification/RCM strategy proved ineffective for the parent natural product because of the difficulties in acylating the sterically encumbered C-13 tertiary alcohol; macrolactionization of a seco-acid was also extensively investigated under various conditions without success. We finally completed the total synthesis of the proposed structure of (-)-lyngbyaloside B by means of a macrolactonization that involves an acyl ketene as the reactive species. However, the NMR spectroscopic data of our synthetic material did not match those of the authentic material, which indicated that the proposed structure must be re-examined. Inspection of the NMR spectroscopic data of the natural product and molecular mechanics calculations led us to postulate that the configuration of the C-10, C-11, and C-13 stereogenic centers had been incorrectly assigned in the proposed structure. Finally, our revised structure of (-)-lyngbyaloside B was unambiguously verified through total synthesis.


Induction of Meal-related Symptoms as a Novel Mechanism of Action of the Duodenal-Jejunal Bypass Sleeve.
Treatment with a duodenal-jejunal bypass sleeve (DJBS) induces clinically significant weight loss, but little is known about the mechanisms of action of this device. The aim of this study was to characterize the mechanisms of action of the DJBS and determine the durability of weight loss and metabolic improvements. We studied a cohort of 19 subjects with severe obesity and type 2 diabetes (baseline body mass index: 43.7±5.3 kg/m). Anthropometry, body composition, blood pressure, biochemical measures, and dietary intake were monitored for 48 weeks after DJBS implantation, and then for 1 year after device removal. Gastric emptying and triglyceride absorption were measured at baseline, 8 weeks after implant, and within 3 weeks of device explant. Visceral sensory function was assessed at baseline, 4 weeks after implant, and within 3 weeks after explant. Significant weight loss (P<0.01) occurred following DJBS placement, with a mean weight reduction of 17.0±6.5% at 48 weeks. The symptom burden following a standardized nutrient challenge was increased after DJBS implantation (P<0.05), returning to baseline after DJBS removal. Neither gastric emptying nor triglyceride absorption changed with the device in situ. A significant reduction in energy intake was observed [baseline: 7703±2978 kJ (1841±712 kcal), 24 weeks: 4824±2259 kJ (1153±540 kcal), and 48 weeks: 4474±1468 kJ (1069±351 kcal)]. After 1 year, anthropometry remained significantly improved, but there was no durable impact on metabolic outcomes. DJBS treatment resulted in substantial weight loss. Weight loss is related to reduced caloric intake, which seems linked to an augmented upper gastrointestinal symptom response, but not altered fat absorption.


Resistance to activated protein C caused by a factor V gene mutation.
Each year, approximately one in 1000 individuals suffers from venous thromboembolism. The pathogenesis of the disease is multifactorial and a thrombotic event is the result of a combination of genetic and circumstantial risk factors. Until recently, genetic defects could only be identified in a minority of thrombosis patients. The discovery of inherited resistance to activated protein C as a risk factor for thrombosis changed the situation for the better. In Western countries, activated protein C resistance is found in 20% to 60% of patients with thrombosis. Activated protein C resistance is caused by a single point mutation in the Factor V gene, leading to replacement of Arg(R)506 in the activated protein C cleavage site of Factor V with a Gln(Q). As a result, the activated protein C-mediated cleavage and inhibition of mutated Factor V (FV:Q506) is impaired, which leads to increased thrombin generation, a hypercoagulable state, and a life-long increased risk of thrombosis.


All-Cause Mortality and Its Predictors in Haemato-Oncology Patients with Febrile Neutropenia.
Febrile neutropenia (FN) is one of the most important life-threatening complications in haemato-oncology. Our objective was to report all-cause mortality rates in patients ill with a hematological malignancy (HM) hospitalized with a first FN episode and to identify predictors for mortality. We conducted a historical retrospective cohort study of consecutive patients with an HM, >18 years of age, admitted between January 2012 and August 2018 for a first episode of FN. Data on all-cause mortality 12 months after admission for FN were obtained. The Kaplan-Meier curve was used to describe mortality during the follow-up period. Univariate and multivariable analyses identified predictors for 1,3 and 12-month mortality. One hundred and fifty-eight patients (mean age 69.5, 49.4% males) were included. Overall, 54 patients died (15.8%, 25.9%, and 34.1% died after 1, 3, and 12 months, respectively). Lower serum albumin, higher serum gamma-glutamyl transferase (GGT), lower estimated glomerular filtration rate (eGFR), older age, higher temperature, and lower absolute lymphocyte count at admission were independent predictors of all-cause mortality after 12 months. Further studies are needed to confirm our results and identify therapeutic strategies to improve survival.


Macrophages as target cells for Mayaro virus infection: involvement of reactive oxygen species in the inflammatory response during virus replication.
Alphaviruses among the viruses that cause arthritis, consisting in a public health problem worldwide by causing localized outbreaks, as well as large epidemics in humans. Interestingly, while the Old World alphaviruses are arthritogenic, the New World alphaviruses cause encephalitis. One exception is Mayaro virus (MAYV), which circulates exclusively in South America but causes arthralgia and is phylogenetically related to the Old World alphaviruses. Although MAYV-induced arthritis in humans is well documented, the molecular and cellular factors that contribute to its pathogenesis are completely unknown. In this study, we demonstrated for the first time that macrophages, key players in arthritis development, are target cells for MAYV infection, which leads to cell death through apoptosis. We showed that MAYV replication in macrophage induced the expression of TNF, a cytokine that would contribute to pathogenesis of MAYV fever, since TNF promotes an inflammatory profile characteristic of arthritis. We also found a significant increase in the production of reactive oxygen species (ROS) at early times of infection, which coincides with the peak of virus replication and precedes TNF secretion. Treatment of the cells with antioxidant agents just after infection completely abolished TNF secretion, indicating an involvement of ROS in inflammation induced during MAYV infection.


Cavity-directed radiosurgery as adjuvant therapy after resection of a brain metastasis.
As a strategy to delay or avoid whole-brain radiotherapy (WBRT) after resection of a brain metastasis, the authors used high-resolution MR imaging and cavity-directed radiosurgery for the detection and treatment of further metastases. Between April 2001 and October 2009, 112 resection cavities in 106 patients with no prior WBRT were treated using radiosurgery directed to the tumor cavity and for any synchronous brain metastases detected on high-resolution MR imaging at the time of radiosurgical planning. A median dose of 17 Gy to the 50% isodose line was prescribed to the gross tumor volume, defined as the rim of enhancement around the resection cavity. Patients were followed up via serial imaging, and new brain metastases were generally treated using additional radiosurgery, with salvage WBRT typically reserved for local treatment failure at a resection cavity, numerous failures, or failures occurring at short time intervals. Local and distant treatment failures were determined based on imaging results. Kaplan-Meier curves were generated to estimate local and distant treatment failure rates, overall survival, neurological cause-specific survival, and time delay to salvage WBRT. Radiosurgery was delivered to the resection cavity alone in 57.5% of patients, whereas 24.5% of patients also received treatment for 1 synchronous metastasis, 11.3% also received treatment for 2 synchronous metastases, and 6.6% also received treatment for 3-10 additional lesions. The median overall survival was 10.9 months. Overall survival at 1 year was 46.8%. The local tumor control rate at 1 year was 80.3%. The disease control rate in distant regions of the brain at 1 year was 35.4%, with a median time of 6.9 months to distant failure. Thirty-nine of 106 patients eventually received salvage WBRT, and the median time to salvage WBRT was 12.6 months. Kaplan-Meier estimates showed that the rate of requisite WBRT at 1 year was 45.9%. Neurological cause-specific survival at 1 year was 50.1%. Leptomeningeal failure occurred in 8 patients. One patient had treatment failure within the resection tract. Seven patients required reoperation: 2 for resection cavity recurrence, 3 for radiation necrosis, 1 for hydrocephalus, and 1 for a CSF cutaneous fistula. On multivariate analysis, a preoperative tumor diameter > 3 cm was predictive of local treatment failure. Cavity-directed radiosurgery combined with high-resolution MR imaging detection and radiosurgical treatment of synchronous brain metastases is an effective strategy for delaying and even foregoing WBRT in most patients. This technique provides acceptable local disease control, although distant treatment failure remains significant.


Prevalence of arterial hypertension among Seventh-Day Adventists of the São Paulo state capital and inner area.
Inadequate life habits are known to favor hypertension, and Adventists recommend healthy life habits. To assess the prevalence of hypertension among Seventh-Day Adventists from the inner São Paulo state and São Paulo state capital. This study assessed 264 Adventists (mean age, 41.17 ± 15.27 years; women, 59.8%) with a high religiosity level assessed by use of the Duke University Religion Index. Blood pressure was measured with a validated automatic device. The significance level adopted was p < 0.05. The total prevalence of hypertension was 22.7% (27.4% in the inner state and 15% in the capital). The Adventists from the capital differed from those of inner state as follows (p < 0.05), respectively: higher education (62% vs 36.6%); employed by a third party (44%) vs self-employed (40.9%); family income (8.39 ± 6.20 vs 4.59 ± 4.75 minimum wages); individual income (4.54 ± 5.34 vs 6.35 ± 48; couple responsible for family income (35% vs 39.6%); vegetarianism (11% vs 3%); blood pressure (115.38 ± 16.52/68.74 ± 8.94 vs 123.66 ± 19.62/74.88 ± 11.85 mmHg); white ethnicity (65% vs 81.1%); married (53% vs 68.9%); lower tangible support in the social aspect (15.7 ± 5.41 vs 16.9 ± 4.32); and recalling the last time one's blood pressure was measured (65% vs 48.8%). On multivariate analysis, hypertension associated with the following: 1) vegetarianism (OR 0.051; 95% CI: 0.004-0.681); 2) educational level (OR 5.317; 95% CI: 1.674-16.893); 3) recalling the last time one's blood pressure was measured (OR 2.725; 95% CI: 1.275-5.821); 4) being retired (OR 8.846; 95% CI: 1.406-55.668); and 5) being responsible for family income (OR 0.422; 95% CI: 0.189-0.942). The prevalence of hypertension among Adventists was lower as compared with that reported in Brazilian studies, and it was lower in the São Paulo state capital as compared with that in the inner São Paulo state, possibly because of the better socioeconomic conditions and life habits of the former.


Singing-related activity in anterior forebrain of male zebra finches reflects courtship motivation for target females.
A critical function of singing by male songbirds is to attract a female mate. Previous studies have suggested that the anterior forebrain system is involved in this courtship behavior. Neural activity in this system, including the striatal Area X, is strikingly dependent on the function of male singing. When males sing to attract a female bird rather than while alone, less variable neural activity results in less variable song spectral features, which may be attractive to the female. These characteristics of neural activity and singing thus may reflect a male's motivation for courtship. Here, we compared the variability of neural activity and song features between courtship singing directed to a female with whom a male had previously formed a pair-bond or to other females. Surprisingly, across all units, there was no clear tendency for a difference in variability of neural activity or song features between courtship of paired females, nonpaired females, or dummy females. However, across the population of recordings, there was a significant relationship between the relative variability of syllable frequency and neural activity: when syllable frequency was less variable to paired than nonpaired females, neural activity was also less variable (and vice-versa). These results show that the lower variability of neural activity and syllable frequency during directed singing is not a binary distinction from undirected singing, but can vary in intensity, possibly related to the relative preference of a male for his singing target.


Effect of glucagon on zinc excretion in anesthetized dogs.
The present studies were performed on anesthetized dogs to determine whether urinary zinc excretion is altered when plasma glucagon concentration is elevated acutely. Glucagon infusion (5 ng.kg-1.min-1) adequate to double base-line plasma glucagon significantly increased zinc excretion in a reversible manner; plasma zinc concentration was unchanged. A larger dose of glucagon (50 ng.kg-1.min-1) produced no significantly greater effect. Clearance experiments using 65Zn to measure ultrafilterable zinc concentration revealed no change in ultrafilterable zinc during glucagon infusion. In a third series of experiments, acetylcholine (25 micrograms/min) was infused into one renal artery while zinc excretion and renal function of the two kidneys were compared. Acetylcholine did not significantly alter zinc excretion despite large changes in sodium and water excretion. The changes in zinc excretion observed in all experiments correlated well with glomerular filtration rate changes, but additional contributions of changes in tubular handling of zinc cannot be ruled out.


Prediction of nonsentinel lymph node metastasis in acral melanoma with positive sentinel lymph nodes.
Metastasis in a nonsentinel lymph node (non-SLN) is an unfavorable independent prognostic factor in cutaneous melanoma (CM). Recent data did suggest potential value of completion lymph node dissection (CLND) in CM patients with non-SLN metastasis. Prediction of non-SLN metastasis assists clinicians in deciding on adjuvant therapy without CLND. We analyzed risk factors and developed a prediction model for non-SLN status in acral melanoma (AM). This retrospective study enrolled 656 cases of melanoma who underwent sentinel lymph node biopsy at Fudan University Shanghai Cancer Center from 2009 to 2017. We identified 81 SLN + AM patients who underwent CLND. Clinicopathologic data, including SLN tumor burden and non-SLN status were examined with Cox and Logistics regression models. Ulceration, Clark level, number of deposits in the SLN (NumDep) and maximum size of deposits (MaxSize) are independent risk factors associated with non-SLN metastases. We developed a scoring system that combines ulceration, the cutoff values of Clark level V, MaxSize of 2 mm, and NumDep of 5 to predict non-SLN metastasis with an efficiency of 85.2% and 100% positive predictive value in the high-rank group (scores of 17-24). A scoring system that included ulceration, Clark level, MaxSize, and NumDep is reliable and effective for predicting non-SLN metastasis in SLN-positive AM.


Effects of environmental temperature and dietary zinc on egg production performance, egg quality and antioxidant status and expression of heat-shock proteins in tissues of broiler breeders.
To investigate the effects of environmental temperature and dietary Zn on egg production performance, egg quality and antioxidant status, as well as expression of heat-shock proteins (HSP) in tissues, of laying broiler breeders, we used a completely randomised design with a 2×3 factorial arrangement of treatments. The two environmental temperatures were normal (21±1°C, NT) and high (32±1°C, HT). The three dietary Zn sources were a Zn-unsupplemented basal diet (CON), and the basal diet supplemented with 110 mg Zn/kg as either the inorganic Zn sulphate (iZn) or the organic Zn proteinate with a moderate chelation strength (oZn). HT decreased (P<0·002) egg weight, laying rate, eggshell strength, thickness and weight, but increased (P≤0·05) rectal temperature, broken egg rate, misshapen egg rate, feed:egg ratio, Cu Zn superoxide dismutase activities in liver and pancreas, as well as metallothionein (MT) level in pancreas, and HSP70 mRNA levels in liver and pancreas of laying broiler breeders. Broiler breeders fed the oZn diet had higher (P<0·04) Zn content in the liver, as well as MT levels in the liver and pancreas, compared with those fed the CON diet. Under HT, broiler breeders fed the oZn diet had higher (P<0·05) Zn content in the pancreas compared with those fed the iZn and CON diets. The results from this study indicated that HT impaired egg production performance and eggshell quality possibly because of the disturbed redox balance and HSP homoeostasis, whereas the oZn is more available than the iZn for pancreatic Zn of heat-stressed laying broiler breeders.