Ion concentrations in nasal airway surface liquid: a prediction model for the identification of cystic fibrosis carriers.

BACKGROUND: Cystic fibrosis (CF) carriers seem to have a higher risk to develop chronic rhino-sinusitis (CRS), although the full underlying mechanisms are unknown. Ion concentrations in nasal airway surface liquid (ASL) may be influenced by the heterozygosity for CF gene mutation, with possible impacts on the development of CRS. METHODS: A cheap and feasible standardized technique was designed to measure the ion levels in nasal ASL. With this purpose we collected, under basal conditions, samples from the nasal cavity of 165 adults: 14 homozygous for CF, 83 carriers and 68 healthy controls. Sodium (Na) and Chlorine (Cl) concentrations were then evaluated among different groups. RESULTS: Statistical analysis revealed a significant difference of Na and Cl values between controls and carriers and between controls and homozygotes. Receiver operating characteristic (ROC) curves and derived indicators (Youden's index and Area Under the Curve, AUC) were used to further evaluate the diagnostic capability of Na and Cl concentrations to differentiate heterozygotes from controls. ROC curves demonstrated that the optimal diagnostic cut-off value of Na is at 124, and the optimal cut-off value of Cl is at 103,2. CONCLUSION: ASL sampling can be considered a new diagnostic tool for providing quantitative information on nasal ion composition. According to our findings, Na and Cl concentrations of nasal ASL could represent a useful tool to assess heterozygotes and healthy controls.

Application of SNPs for population genetics of nonmodel organisms: new opportunities and challenges.

Recent improvements in the speed, cost and accuracy of next generation sequencing are revolutionizing the discovery of single nucleotide polymorphisms (SNPs). SNPs are increasingly being used as an addition to the molecular ecology toolkit in nonmodel organisms, but their efficient use remains challenging. Here, we discuss common issues when employing SNP markers, including the high numbers of markers typically employed, the effects of ascertainment bias and the inclusion of nonneutral loci in a marker panel. We provide a critique of considerations specifically associated with the application and population genetic analysis of SNPs in nonmodel taxa, focusing specifically on some of the most commonly applied methods.

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 December 2009-31 January 2010.

This article documents the addition of 220 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Allanblackia floribunda, Amblyraja radiata, Bactrocera cucurbitae, Brachycaudus helichrysi, Calopogonium mucunoides, Dissodactylus primitivus, Elodea canadensis, Ephydatia fluviatilis, Galapaganus howdenae howdenae, Hoplostethus atlanticus, Ischnura elegans, Larimichthys polyactis, Opheodrys vernalis, Pelteobagrus fulvidraco, Phragmidium violaceum, Pistacia vera, and Thunnus thynnus. These loci were cross-tested on the following species: Allanblackia gabonensis, Allanblackia stanerana, Neoceratitis cyanescens, Dacus ciliatus, Dacus demmerezi, Bactrocera zonata, Ceratitis capitata, Ceratitis rosa, Ceratits catoirii, Dacus punctatifrons, Ephydatia mülleri, Spongilla lacustris, Geodia cydonium, Axinella sp., Ischnura graellsii, Ischnura ramburii, Ischnura pumilio, Pistacia integerrima and Pistacia terebinthus.

Characterization of microsatellite loci in the subsocial spider Stegodyphus lineatus (Araneae: Eresidae).

Stegodyphus lineatus spiders live in groups consisting of closely related individuals. There appears to be no discrimination against related individuals as mates but females mate multiply, despite the fact that matings are shown to carry a cost. We have developed eight polymorphic dinucleotide microsatellite markers that allow us to assess levels of heterozygosity and relatedness among individuals of this species. These molecular markers are likely to prove highly effective tools for estimating levels of inbreeding and thus allow us to test hypotheses about the relationships between social structure, mating strategies and inbreeding avoidance.

Human T-cell lymphotropic virus type I (HTLV-I) confirmed by PCR-Southern blot and sequencing analysis in a woman with tropical spastic paraparesis.

Human T-cell lymphotropic virus type I (HTLV-I) is a human retrovirus and the aetiological agent of a progressive neurological disease called tropical spastic paraparesis/HTLV-I-associated myelopathy (TSP/HAM), as confirmed by evidence accumulated in HTLV-I seroprevalence studies. TSP/HAM is rarely diagnosed in Italy, given the low prevalence of HTLV-I in the population. TSP/HAM begins insidiously in the fourth decade, mainly with spastic paraparesis of the lower extremities and positive Babinski reflex, as well as interfering with bowel and bladder functions. In this study we report the clinical, virological and haemato chemical data of a 54-year-old woman, born in the Ivory Cost, with symptoms suggestive of TSP. The presence of HTLV-I infection was demonstrated by the detection of antibodies in serum and in cerebrospinal fluid by immunoenzymatic assay and Western blot analysis. In addition, viral isolation was carried out in peripheral blood cells, and the presence of HTLV-I proviral DNA was confirmed by polymerase chain reaction/Southern blot and sequencing analysis. According to our results, HTLV-I testing might be useful when TSP/HAM is suspected.

Cryopreserved human hepatocytes from cell bank: in vitro function and clinical application.

UNLABELLED: We Aimed to analyze the in vitro function of isolated and cryopreserved human hepatocytes (CHH) from a cell bank and to define their potential clinical application in a bioartificial liver (BAL) device. METHODS: Over 24 months, 103 not transplantable livers were utilized for human hepatocytes isolation and cryopreservation. Hepatocytes isolated by collagenase were analyzed for yield, viability, diazepam metabolism, and production of human albumin after isolation and cryopreservation in LN(2). RESULTS: The causes for refusal for transplantation were macrosteatosis >60%, ischemic damage due to donor hypotension, and nonviral cirrhosis in 60%, 11%, and 8%, respectively. Cell yields averaged 7 million hepatocytes per gram of liver of mean viability of 80% +/- 13%. The viability of CHH after thawing averaged 50%. Thawed hepatocytes showed diazepam metabolism, and human albumin synthesis comparable to fresh cells. CHH were utilized as the biological component of a BAL for temporary support as three applications of two patients affected by fulminant hepatic failure awaiting urgent transplant. Ten to 13 billion viable CHH were loaded into each BAL. Liver function showed bilirubin and ammonia reduction at the end of each treatment. One patient was successfully bridged to emergency OLTx after one BAL; in the second case there was spontaneous recovery of liver function after two BAL. CONCLUSIONS: Recovery of donor human livers unwanted for transplantation allowed isolation and cryopreservation of viable and functionally active human hepatocytes, which have been banked and successfully used for clinical applications of a BAL device.

Inhibition of neutrophil responses by cyclosporin A. An insight into molecular mechanisms.

OBJECTIVE: Cyclosporin A (CsA) is an effective agent in rheumatoid arthritis (RA), slowing joint damage progression. Its therapeutic effect on T lymphocytes has been studied extensively, but there is little information available about neutrophils, the cells responsible for a substantial proportion of inflammation. A study was performed to investigate the in vitro effects of CsA on neutrophil functions triggered by several agonists and determine whether the drug could counteract the binding of formyl-methionyl-leucyl-phenylalanine (fMLP) to its receptor and/or modulate changes in the intracellular Ca(2+) concentration ([Ca(2+)]i). METHODS: CsA was added to neutrophils 5-50 min before the incubation steps for neutrophil function assays (chemotaxis, superoxide anion production, lysozyme release), calcium measurements and receptor binding experiments. RESULTS: CsA appeared to be particularly effective in lowering chemotaxis, superoxide anion production and lysozyme release induced by different agonists. However, it did not significantly affect either basal or agonist-stimulated neutrophil [Ca(2+)]i and the interaction between fMLP and its receptor. CONCLUSIONS: Because of its in vitro inhibition of neutrophil functions, CsA appears to have considerable potential as an anti-inflammatory drug. Moreover, as it is also a potent immunosuppressive agent, it may reduce the progression of joint damage in RA. More work remains to be done to clarify the molecular mechanism of CsA action on neutrophils.

Two for-Met-Leu-Phe-OMe analogues trigger selective neutrophil responses. A differential effect on cytosolic free Ca2+.

For-Thp-Leu-Ain-OMe and for-Met-delta(z)Leu-Phe-OMe are two conformationally restricted fMLP-OMe analogues able to discriminate between different biological responses of human neutrophils. In this paper, we demonstrate that the former peptide, which evokes only chemotaxis, does not alter human neutrophil Ca2+ levels. In contrast, for-Met-delta(z)Leu-Phe-OMe, which induces superoxide anion release and degranulation but not chemotaxis, significantly increases the cation concentration. The chelation of Ca2+ in both extracellular and intracellular media abolishes O2- production triggered by for-Met-delta(z)Leu-Phe-OMe, while the same procedure does not affect neutrophil chemotaxis towards for-Thp-Leu-Ain-OMe. We therefore suggest that chemotaxis, unlike superoxide anion release, is independent of Ca2+ enhancement in human neutrophils.

[Dopamine as a regulator of natriuresis. Experimental bases and their implications]. / La dopamina come regolatore della natriuria. Basi sperimentali e loro implicazioni.

Renal effects of dopamine (DA) infused i.v. in a subpressor dose (0.1 microgram. kg.-1 min-1) were investigated in healthy human subjects during steady hypotonic polyuria. In each experiment four clearance (cl.) periods of 15 min were performed; DA was administered during the second and the third cl. periods. The glomerular filtration rate and renal effective plasma flow were estimated as endogenous creatinine and PAH clearances. Moreover the following variables were evaluated: a) sodium total and sodium isosmotic reabsorption as a % of sodium filtered load (s.f.l.); b) sodium anisosmotic reabsorption as a % of sodium distal load (s.d.l.); c) total renal vascular resistance (RT), arteriolar afferent (RA) and efferent (RE) resistances. Three groups of experiments were performed: A) in hydro-saline retention by DOCA pretreatment (24 subjects); B) in hydro-saline depletion by natriuretic pretreatment (20 subjects); C) in hydro-saline depletion and beta-adrenergic blockade induced by natriuretic and propranolol pretreatments (9 subjects). In A experimental condition DA significantly increased renal plasma flow, glomerular filtration rate, urinary flow, urine sodium concentration and osmolarity; on the contrary DA significantly decreased RT, RA, sodium total and isosmotic reabsorption, % of s.f.l., and sodium anisosmotic reabsorption, % of s.d.l. In B condition no significant changes were observed during DA infusion in either haemodynamic parameters or urinary flow; on the other hand DA significantly blunted urinary osmolarity, urinary sodium concentration and sodium excretion rate suggesting an increased sodium anisosmotic reabsorption (% of s.d.l.). In C condition the vasodilating and hydrosaluretic effects of DA were restored. Our results suggest that DA activates, besides "dopaminic" vascular receptors, the presynaptic facilitory beta-adrenergic receptors in renal adrenergic pathways. Thus the renal action of DA depends on renal adrenergic activity.

[Dopamine, catecholamine receptors, and neurohumoral regulation of renal function]. / Dopamina, recettori catecolaminici e regolazione neuro-ormonale della funzione del rene.

Were studied in healthy human subjects under conditions of hydro-saline retention the intrarenal mechanism underlying the hydronatriuretic effect of Dopamine (DA) and the changes in DA renal effects induced by Sulpiride (S). DA was infused i.v. in a subpressor dose (0,1 microgram/kg . min) during induced hypotonic polyuria. In each experiment four clearance periods of 15 min were performed; DA was administered during the second and third clearance periods. The glomerular filtration rate and renal effective plasma flow were estimated as endogenous creatinine and PAH clearances, respectively. Tubular sodium and potassium reabsorptions were also determined. 1) In the state of hydro-saline retention, renal arteriolar (mainly preglomerular) vasodilation was produced by DA. Moreover, both sodium isosmotic reabsorption as a percentage of sodium filtered load and sodium anisosmotic reabsorption as a percentage of sodium distal load were inhibited. These tubular inhibitions were found to be correlated with the haemodynamic effects of DA. 2) Sulpiride treatment (4,4 mg/kg . day given orally for two days prior to the experiment and 100 mg i.m. 40 min before DA infusion) caused (a) an increase in the hydro-natriuretic response to hydration during control clearance and (b) a decrease in DA haemodynamic effects. An interpretation is proposed accounting for these DA effects as well as for dependence of DA renal effects on the extracellular fluid volume.