Differential white blood cell counts in rabbits: a comparison of the Advia 2120 and a manual method

We evaluated the performance of the Advia 2120 (Siemens) differential leukocyte count (A-Diff) compared to the manual method (M-Diff) in rabbits. EDTA-anticoagulated blood samples collected for diagnostic purposes were analyzed within 6 h of collection. The M-Diff was performed blindly by 2 observers on blood smears by counting 200 cells. We initially included 117 samples; 25 samples were excluded because of suboptimal gating of leukocytes in the Advia peroxidase cytogram or poor blood smear quality. The correlation between the A-Diff and M-Diff was very high for heterophils (r = 0.924, p < 0.001) and lymphocytes (r = 0.903, p < 0.001), high for basophils (r = 0.823, p < 0.001), moderate for monocytes (r = 0.645, p < 0.001), and low for eosinophils (r = 0.336, p = 0.001). The Passing–Bablok regression analyses revealed a small-to-moderate constant error for lymphocytes and a slight constant error for basophils. Small proportional errors were detected for heterophils, lymphocytes, and eosinophils. The Bland–Altman analyses revealed that the Advia significantly underestimates heterophils and overestimates lymphocytes compared to M-Diff. The biases for the other leukocytes were minimal and likely clinical insignificant; however, our results, particularly for eosinophils, should be interpreted cautiously given the observed low percentages in our samples. Given the observed biases in heterophil and lymphocyte percentages in the Advia 2120 CBC results in rabbits, method-specific reference intervals should be used. The Advia can recognize leporine basophils. Evaluation of blood smears is still recommended to investigate abnormal results and erroneous cytograms reported by the Advia

Effect of the Counterion on Circularly Polarized Luminescence of Europium(III) and Samarium(III) Complexes

Each enantiopure europium(III) and samarium(III) nitrate and triflate complex of the ligand L, with L = N,N'-bis(2-pyridylmethylidene)-1,2-(R,R + S,S)-cyclohexanediamine ([LnL(tta)2]·NO3 and [LnL(tta)2(H2O)]·CF3SO3, where tta = 2-thenoyltrifluoroacetylacetonate) has been synthesized and characterized from a spectroscopic point of view, using a chiroptical technique such as electronic circular dichroism (ECD) and circularly polarized luminescence (CPL). In all cases, both ligands are capable of sensitizing the luminescence of both metal ions upon absorption of light around 280 and 350 nm. Despite small differences in the total luminescence (TL) and ECD spectra, the CPL activity of the complexes is strongly influenced by a concurrent effect of the solvent and counterion. This particularly applies to europium(III) complexes where the CPL spectra in acetonitrile can be described as a weighed linear combination of the CPL spectra in dichloromethane and methanol, which show nearly opposite signatures when their ligand stereochemistries are the same. This phenomenon could be related to the presence of equilibria interconverting solvated, anion-coordinated complexes and isomers differing by the relative orientation of the tta ligands. The difference between some bond lengths (M-N bonds, in particular) in the different species could be at the basis of such an unusual CPL activity

Effect of the Heteroaromatic Antenna on the Binding of Chiral Eu(III) Complexes to Bovine Serum Albumin

The cationic enantiopure R) and luminescent Eu(III) complex [Eu(bisoQcd)(H2O)(2)] OTf (with bisoQcd = N,N'-bis(2-isaquinolinmethyl)-trarts-1,2diaminocyclohexane N/N1 -diacetate and OTf = triflate) was synthesized and characterized. At physiological pH, the 1:1 [Eu(bisoQcd)(H2O)(2)](+) species, possessing two water molecules in the inner coordination sphere, is largely dominant. The interaction with bovine serum albumin (BSA) was studied by means of several experimental techniques, such as luminescence spectroscopy, isothermal titration calorimeti-y (ITC), molecular docking (MD), and molecular dynamics simulations M11.-.)S). In this direction, a ligand competition study was also performed by using three clinically established drugs (i.e., ibuprofen, warfarin, and digito)cin). The nature of this interaction is strongly affected by the type of the involved heteroaromatic antenna in the complexes. In fact, the presence of isoqiiinolirie rings drives the corresponding complex toward the protein superficial area containing the tryptophan residue 134 (Trp134). As the main consequence, the metal center undergoes the loss of one water molecule upon interaction with the side chain of a glutamic acid residue. On the other hand, the similar complex containing pyridine rings f[Eti(bpcd)(H2O)(2)]Cl with bpcd = N,N'-bis(2-pytidylmethyl)-trans-1,2-diaminocyclohexane N,N'-diacetate)interacts more weakly with the protein in a different superficial cavity, without losing the coordinated water molecules

Eu(iii) and Tb(iii) complexes of 6-fold coordinating ligands showing high affinity for the hydrogen carbonate ion: A spectroscopic and thermodynamic study

In the present contribution, four classes of Ln(iii) complexes (Ln = Eu and Tb) have been synthesized and characterized in aqueous solution. They differ by charge, Ln(bpcd)+ [bpcd2- = N,N'-bis(2-pyridylmethyl)-trans-1,2-diaminocyclohexane N,N'-diacetate] and Ln(bQcd)+ (bQcd2- = N,N'-bis(2-quinolinmethyl)-trans-1,2-diaminocyclohexane N,N'-diacetate) being positively charged and Ln(PyC3A) (PyC3A3- = N-picolyl-N,N',N'-trans-l,2-cyclohexylenediaminetriacetate) and Ln(QC3A) (QC3A3- = N-quinolyl-N,N',N'-trans-l,2-cyclohexylenediaminetriacetate) being neutral. Combined DFT, spectrophotometric and potentiometric studies reveal the presence, under physiological conditions (pH 7.4), of a couple of equally and highly stable isomers differing by the stereochemistry of the ligands (trans-N,N and trans-O,O for bpcd2- and bQcd2-; trans-O,O and trans-N,O for PyC3A3- and QC3A3-). Their high log\u2009\u3b2 values (9.97 &lt; log\u2009\u3b2 &lt; 15.68), the presence of an efficient antenna effect and the strong increase of the Ln(iii) luminescence intensity as a function of the hydrogen carbonate concentration in physiological solution, render these complexes as very promising optical probes for a selective detection of HCO3-in cellulo or in extracellular fluid. This particularly applies to the cationic Eu(bpcd)+, Tb(bpcd)+ and Eu(bQcd)+ complexes, which are capable of guesting up to two hydrogen carbonate anions in the inner coordination sphere of the metal ion, so that they show an unprecedented affinity towards HCO3- (log\u2009K for the formation of the adduct in the 4.6-5.9 range)

Optical properties of Eu(III) and Tb(III) complexes with pyridine- and quinoline- based ligands under high hydrostatic pressure

The spectroscopy of nitrate complexes of Eu(III) and Tb(III) with chiral and racemic imine-based [L1 = (N,N'-bis (2-pyridylmethylidene)-1,2-(R,R + S,S)-cyclohexanediamine) and L3 = N, N'-bis(2-quinolylmethylidene)-1,2-(R,R + S,S)-cyclohexanediamine] and amine-based [L2 = N,N'-bis(2-pyridylmethyl)-1,2-(R,R + S,S)-cyclohexanediamine) and L4 = N,N'-bis(2-quinolylmethyl)-1,2-(R,R + S,S)-cyclohexanediamine] ligands has been studied under high hydrostatic pressure (above 100 kbar). With the increasing pressure, a reduction of the Tb(III) and Eu(III) luminescence intensity is detected for all the complexes, whilst a significant reduction of the Tb(III) and Eu(III) excited state lifetimes has been observed for all Tb-based complexes [L1Tb(NO3)(3) -&gt; L4Tb(NO3)(3)] and only for the Eu(III) complexes containing the imine-based ligands [L1Eu(NO3)(3) and L3Eu(NO3)(3)]. This behavior has been rationalized taking into account two main aspects: i) the relative position of the energy levels of the ligands and the metal ions and ii) the change of these position upon compression DFT calculations have been also performed to elucidate the nature of the orbitals involved in the UV electronic absorption transitions (NTO orbitals) upstream of the energy transfer process to the metal ion

Ectodysplasin signalling deficiency in mouse models of Hypohidrotic Ectodermal Dysplasia leads to middle ear and nasal pathology

Hypohidrotic ectodermal dysplasia (HED) results from mutation of the EDA, EDAR or EDARADD genes and is characterized by reduced or absent eccrine sweat glands, hair follicles and teeth, and defective formation of salivary, mammary and craniofacial glands. Mouse models with HED also carry Eda, Edar or Edaradd mutations and have defects that map to the same structures. Patients with HED have ear, nose and throat disease, but this has not been investigated in mice bearing comparable genetic mutations. We report that otitis media, rhinitis and nasopharyngitis occur at high frequency in Eda and Edar mutant mice and explore the pathogenic mechanisms related to glandular function, microbial and immune parameters in these lines. Nasopharynx auditory tube glands fail to develop in HED mutant mice and the functional implications include loss of lysozyme secretion, reduced mucociliary clearance and overgrowth of nasal commensal bacteria accompanied by neutrophil exudation. Heavy nasopharynx foreign body load and loss of gland protection alters the auditory tube gating function and the auditory tubes can become pathologically dilated. Accumulation of large foreign body particles in the bulla stimulates granuloma formation. Analysis of immune cell populations and myeloid cell function shows no evidence of overt immune deficiency in HED mutant mice. Our findings using HED mutant mice as a model for the human condition support the idea that ear and nose pathology in HED patients arises as a result of nasal and nasopharyngeal gland deficits, reduced mucociliary clearance and impaired auditory tube gating function underlies the pathological sequelae in the bulla

Nosocomial outbreak of the pandemic Influenza A (H1N1) 2009 in critical hematologic patients during seasonal influenza 2010-2011: detection of oseltamivir resistant variant viruses

BACKGROUND: The pandemic influenza A (H1N1) 2009 (H1N1pdm09) virus infection caused illness and death among people worldwide, particularly in hematologic/oncologic patients because influenza infected individuals can shed virus for prolonged periods, thus increasing the chances for the development of drug-resistant strains such as oseltamivir-resistant (OST-r) variant. METHODS: The aim of our study was to retrospectively evaluate the clinical importance of OST-r variant in circulating strains of the pandemic H1N1pdm09 virus. By means of RT-PCR and Sanger sequencing we analysed the presence of OST-r variant in 76 H1N1pdm09 laboratory-confirmed cases, hospitalized at the hematologic/oncologic ward at Spedali Civili of Brescia –Italy. RESULTS: Out of 76 hospitalized hematologic/oncologic patients, 23 patients (30.2%) were infected by H1N1pdm09 virus. Further investigation revealed that 3 patients were positive for the OST-r variant carrying the H275Y mutation. All the 23 infected patients were immuno-compromised, and were under treatment or had been treated previously with oseltamivir. Three patients died (13%) after admission to intensive care unit and only one of them developed H275Y mutation. CONCLUSIONS: Our retrospective observational study shows that pandemic influenza A (H1N1) 2009 virus can cause significant morbidity and even mortality in hematologic/oncologic patients and confirms the high rate of nosocomial transmission of pandemic H1N1pdm09 virus in these critical subjects. Indeed, the reduction in host defences in these hospitalized patients favoured the prolonged use of antiviral therapy and permitted the development of OST-r strain. Strategies as diagnostic vigilance, early isolation of patients and seasonal influenza A(H1N1) vaccination may prevent transmission of influenza in high risk individuals