Deep sequencing reveals persistence of cell-associated mumps vaccine virus in chronic encephalitis.

Routine childhood vaccination against measles, mumps and rubella has virtually abolished virus-related morbidity and mortality. Notwithstanding this, we describe here devastating neurological complications associated with the detection of live-attenuated mumps virus Jeryl Lynn (MuV(JL5)) in the brain of a child who had undergone successful allogeneic transplantation for severe combined immunodeficiency (SCID). This is the first confirmed report of MuV(JL5) associated with chronic encephalitis and highlights the need to exclude immunodeficient individuals from immunisation with live-attenuated vaccines. The diagnosis was only possible by deep sequencing of the brain biopsy. Sequence comparison of the vaccine batch to the MuV(JL5) isolated from brain identified biased hypermutation, particularly in the matrix gene, similar to those found in measles from cases of SSPE. The findings provide unique insights into the pathogenesis of paramyxovirus brain infections

Determination of nutrient salts by automatic methods both in seawater and brackish water: the phosphate blank

9 páginas, 2 tablas, 2 figurasThe main inconvenience in determining nutrients in seawater by automatic methods is simply solved: the preparation of a suitable blank which corrects the effect of the refractive index change on the recorded signal. Two procedures are proposed, one physical (a simple equation to estimate the effect) and the other chemical (removal of the dissolved phosphorus with ferric hydroxide).Support for this work came from CICYT (MAR88-0245 project) and Conselleria de Pesca de la Xunta de GaliciaPeer reviewe

Stereo-Specific Modulation of the Extracellular Calcium-Sensing Receptor in Colon Cancer Cells

Pharmacological allosteric agonists (calcimimetics) of the extracellular calcium-sensing receptor (CaSR) have substantial gastro-intestinal side effects and induce the expression of inflammatory markers in colon cancer cells. Here, we compared the effects of both CaSR-specific (R enantiomers) and -unspecific (S enantiomers) enantiomers of a calcimimetic (NPS 568) and a calcilytic (allosteric CaSR antagonists; NPS 2143) to prove that these effects are indeed mediated via the CaSR, rather than via off-target effects, e.g., on β-adrenoceptors or calcium channels, of these drugs. The unspecific S enantiomer of NPS 2143 and NPS S-2143 was prepared using synthetic chemistry and characterized using crystallography. NPS S-2143 was then tested in HEK-293 cells stably transfected with the human CaSR (HEK-CaSR), where it did not inhibit CaSR-mediated intracellular Ca2+ signals, as expected. HT29 colon cancer cells transfected with the CaSR were treated with both enantiomers of NPS 568 and NPS 2143 alone or in combination, and the expression of CaSR and the pro-inflammatory cytokine interleukin 8 (IL-8) was measured by RT-qPCR and ELISA. Only the CaSR-selective enantiomers of the calcimimetic NPS 568 and NPS 2143 were able to modulate CaSR and IL-8 expression. We proved that pro-inflammatory effects in colon cancer cells are indeed mediated through CaSR activation. The non-CaSR selective enantiomer NPS S-2143 will be a valuable tool for investigations in CaSR-mediated processes

The calcium-sensing receptor in physiology and in calcitropic and noncalcitropic diseases

The Ca2+-sensing receptor (CaSR) is a dimeric family C G protein-coupled receptor that is expressed in calcitropic tissues such as the parathyroid glands and the kidneys and signals via G proteins and β-arrestin. The CaSR has a pivotal role in bone and mineral metabolism, as it regulates parathyroid hormone secretion, urinary Ca2+ excretion, skeletal development and lactation. The importance of the CaSR for these calcitropic processes is highlighted by loss-of-function and gain-of-function CaSR mutations that cause familial hypocalciuric hypercalcaemia and autosomal dominant hypocalcaemia, respectively, and also by the fact that alterations in parathyroid CaSR expression contribute to the pathogenesis of primary and secondary hyperparathyroidism. Moreover, the CaSR is an established therapeutic target for hyperparathyroid disorders. The CaSR is also expressed in organs not involved in Ca2+ homeostasis: it has noncalcitropic roles in lung and neuronal development, vascular tone, gastrointestinal nutrient sensing, wound healing and secretion of insulin and enteroendocrine hormones. Furthermore, the abnormal expression or function of the CaSR is implicated in cardiovascular and neurological diseases, as well as in asthma, and the CaSR is reported to protect against colorectal cancer and neuroblastoma but increase the malignant potential of prostate and breast cancers

Calcium-sensing receptor antagonists abrogate airways hyperresponsiveness and inflammation in allergic asthma

Airways hyperresponsiveness and inflammation are fundamental hallmarks of allergic asthma and are accompanied by increases in certain polycations, such as eosinophil cationic protein, whose levels in body fluids correlate with asthma severity. Here we show that polycations, or elevated extracellular calcium activate the human recombinant and native calcium-sensing receptor (CaSR), leading to intracellular calcium mobilization, cAMP breakdown and p38 MAPK phosphorylation in airway smooth muscle (ASM) cells, effects prevented by CaSR antagonists, termed calcilytics. Asthmatic patients and allergen-sensitized mice expressed more CaSR in ASMs than their healthy counterparts. Polycations induced hyperreactivity in mouse bronchi, effect prevented by calcilytics and absent in mice with CaSR ablation from ASM. Calcilytics also reduced airway hyperresponsiveness and inflammation in allergen-sensitized mice in vivo. These data show that a functional CaSR is upregulated in asthmatic ASM and targeted by locally produced polycations to induce hyperresponsiveness and inflammation. Calcilytics may represent a novel, effective asthma therapeutics

Menstrual Variation in Experimental Pain: Correlation with Gonadal Hormones

&lt;i&gt;Background:&lt;/i&gt; The results of studies examining the response to experimental pain during the menstrual cycle are conflicting because of differences in the definitions of the menstrual period, outcome measures and types of experimental pain stimulation. So far, there have been only a few studies correlating experimental pain with the levels of gonadal hormones over the menstrual cycle. Therefore, we assessed the responses to multiple experimental pain stimuli during the menstrual cycle and computed their correlations with the salivary concentrations of the gonadal hormones estrogen and testosterone. &lt;i&gt;Methods:&lt;/i&gt; Twenty-four healthy and regularly menstruating women between 20 and 41 years old took part in the study. Detection thresholds (warmth, cold and electrical current) and pain thresholds (cold, heat, pressure and electrical current) were assessed on days 1, 4, 14 and 22 of the menstrual cycle. In each session, salivary samples were collected for the determination of the physiological estrogen 17β-estradiol, progesterone and testosterone. Progesterone was used exclusively to verify regular menstrual cycling. &lt;i&gt;Results:&lt;/i&gt; Significant variations in pain thresholds for cold, pressure and electrical stimuli were observed over the menstrual cycle with the highest thresholds on day 22, except for the cold pain thresholds, which peaked on day 14. There were no such changes regarding heat pain and all the detection thresholds. The correlations separately computed for each of the 4 days between salivary estrogen as well as testosterone on the one hand and the detection or pain thresholds on the other hand failed to show significant levels, except for the coupling of testosterone and electrical pain thresholds on day 1. &lt;i&gt;Conclusions:&lt;/i&gt; The pain thresholds for all the physical stressors increased after menstruation. The acrophases were located in the follicular (cold pain threshold) or in the luteal phase (pressure and electrical pain thresholds). The results of our correlation analyses indicate only minimal influences of the physiological levels of gonadal hormones on pain sensitivity in women.</jats:p