Quantification of the early pupillary dilation kinetic to assess rod and cone activity.

Rods, cones and melanopsin contribute in various proportions, depending on the stimulus light, to the pupil light response. This study used a first derivative analysis to focus on the quantification of the dynamics of pupillary dilation that immediately follows light-induced pupilloconstriction in order to identify novel parameters that reflect rod and cone activity. In 18 healthy adults, the pupil response to a 1 s blue light stimulus ranging from - 6.0 to 2.65 log cd/m <sup>2</sup> in dark-adapted conditions and to a 1 s blue light stimulus (2.65 log cd/m <sup>2</sup> ) in light-adapted conditions was recorded on a customized pupillometer. Three derivative parameters which describe the 2.75 s following the light onset were quantified: dAMP (maximal amplitude of the positive peak), dLAT (latency of the positive peak), dAUC (area under the curve of the positive peak). We found that dAMP and dAUC but not dLAT have graded responses over a range of light intensities. The maximal positive value of dAMP, representing maximal rate of change of early pupillary dilation phase, occurs at - 1.0 log cd/m <sup>2</sup> and this stimulus intensity appears useful for activating rods and cones. From - 0.5 log cd/m <sup>2</sup> to brighter intensities dAMP and dAUC progressively decrease, reaching negligible values at 2.65 log cd/m <sup>2</sup> indicative of a melanopsin-driven pupil response that masks the contribution from rods and cones to the early phase of pupillary dilation

Determination of Rod and Cone Influence to the Early and Late Dynamic of the Pupillary Light Response.

PURPOSE: This study aims to identify which aspects of the pupil light reflex are most influenced by rods and cones independently by analyzing pupil recordings from different mouse models of photoreceptor deficiency. METHODS: One-month-old wild type (WT), rodless (Rho-/-), coneless (Cnga3-/-), or photoreceptor less (Cnga3-/-; Rho-/- or Gnat1-/-) mice were subjected to brief red and blue light stimuli of increasing intensity. To describe the initial dynamic response to light, the maximal pupillary constriction amplitudes and the derivative curve of the first 3 seconds were determined. To estimate the postillumination phase, the constriction amplitude at 9.5 seconds after light termination was related to the maximal constriction amplitude. RESULTS: Rho-/- mice showed decreased constriction amplitude but more prolonged pupilloconstriction to all blue and red light stimuli compared to wild type mice. Cnga3-/- mice had constriction amplitudes similar to WT however following maximal constriction, the early and rapid dilation to low intensity blue light was decreased. To high intensity blue light, the Cnga3-/- mice demonstrated marked prolongation of the pupillary constriction. Cnga3-/-; Rho-/- mice had no pupil response to red light of low and medium intensity. CONCLUSIONS: From specific gene defective mouse models which selectively voided the rod or cone function, we determined that mouse rod photoreceptors are highly contributing to the pupil response to blue light stimuli but also to low and medium red stimuli. We also observed that cone cells mainly drive the partial rapid dilation of the initial response to low blue light stimuli. Thus photoreceptor dysfunction can be derived from chromatic pupillometry in mouse models

Comparison of presepsin, procalcitonin, interleukin-8 and C-reactive protein in predicting bacteraemia in febrile neutropenic adult patients with haematological malignancies

Bacterial infections represent life-threatening complications in patients with febrile neutropenia (FN). Diagnostic biomarkers of infections may help to differentiate bacteraemia from non-bacteraemia FN. We aimed to evaluate the utility of procalcitonin (PCT), presepsin (PS), C-reactive protein (CRP) and interleukin-8 (IL-8) as biomarkers of bacteraemia in adult FN patients with haematological malignancies. Concentrations of PCT, PS, CRP and IL-8 were prospectively measured in 36 FN episodes experienced by 28 oncohaematological patients. 11 out of 36 episodes were classified as bacteraemia. PCT was the best biomarker to predict bacteraemia with the area under the curve (AUC) ROC of 0,9; specificity 100% and positive predictive value 100%, while the most sensitive was IL-8 (90,9%) with AUC ROC of 0,88 and negative predictive value 95,2%. All patients with PCT concentrations above 1,6 \u3bcg/l had bacteraemia. Patients with IL-8 concentrations superior to 170 pg/ml had a 40 times higher risk for bacteraemia than the ones with lower levels. Patients with PS concentrations superior to 410 pg/ml had 24 times higher risk for bacteraemia than the patients with lower levels. PCT has higher accuracy than CRP, IL-8 and PS in predicting bacteraemia in adult hematologic patients with FN

Paramagnetic Meissner Effect in Multiply-Connected Superconductors

We have measured a paramagnetic Meissner effect in Nb-Al2O3-Nb Josephson junction arrays using a scanning SQUID microscope. The arrays exhibit diamagnetism for some cooling fields and paramagnetism for other cooling fields. The measured mean magnetization is always less than 0.3 flux quantum (in terms of flux per unit cell of the array) for the range of cooling fields investigated. We demonstrate that a new model of magnetic screening, valid for multiply-connected superconductors, reproduces all of the essential features of paramagnetism that we observe and that no exotic mechanism, such as d-wave superconductivity, is needed for paramagnetism.Comment: 4 pages, 3 figures, LaTe

Aging Effect in Ceramic Superconductors

A three-dimensional lattice of the Josephson junctions with a finite self-conductance is employed to model ceramic superconductors. Using Monte Carlo simulations it is shown that the aging disappears in the strong screening limit. In the weeak screening regime aging is present even at low temperatures. For intermediate values of the self-inductance aging occurs at intermediate temperatures interval but is suppressed entirely at high and low temperatures. Our results are in good agreement with experiments.Comment: 5 pages, 5 eps figures, to appear in Physical Review Letter

Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors.

Lentivirus-based gene delivery vectors carrying multiple gene cassettes are powerful tools in gene transfer studies and gene therapy, allowing coexpression of multiple therapeutic factors and, if desired, fluorescent reporters. Current strategies to express transgenes and microRNA (miRNA) clusters from a single vector have certain limitations that affect transgene expression levels and/or vector titers. In this study, we describe a novel vector design that facilitates combined expression of therapeutic RNA- and protein-based antiangiogenic factors as well as a fluorescent reporter from back-to-back RNApolII-driven expression cassettes. This configuration allows effective production of intron-embedded miRNAs that are released upon transduction of target cells. Exploiting such multigenic lentiviral vectors, we demonstrate robust miRNA-directed downregulation of vascular endothelial growth factor (VEGF) expression, leading to reduced angiogenesis, and parallel impairment of angiogenic pathways by codelivering the gene encoding pigment epithelium-derived factor (PEDF). Notably, subretinal injections of lentiviral vectors reveal efficient retinal pigment epithelium-specific gene expression driven by the VMD2 promoter, verifying that multigenic lentiviral vectors can be produced with high titers sufficient for in vivo applications. Altogether, our results suggest the potential applicability of combined miRNA- and protein-encoding lentiviral vectors in antiangiogenic gene therapy, including new combination therapies for amelioration of age-related macular degeneration

Maturation of the Pupil Light Reflex Occurs Until Adulthood in Mice.

With respect to photoreceptor function, it is well known that electroretinogram (ERG) amplitudes decrease with age, but to our knowledge, studies describing age-related changes in the pupil light response (PLR) of mice are lacking. This study recorded the PLR and ERG in C57BL/6 and Sv129S6 wild-type mice at three different ages during early adulthood. Dark- and light-adapted PLR and ERG measurements were performed at 1, 2, and 4 months of age. For PLR measurements, we used either a red (622 nm) or blue (463 nm) light stimulus (500 ms) to stimulate one eye. We selected various light intensities ranging across almost 4 log units and subsequently classified them as low, medium, or high intensity. From the recorded PLR, we selected parameters to quantify the early and late phases of the response such as the baseline pupil size, the maximal constriction amplitude, the maximal velocity, the early partial dilation (area under the curve of the positive peak of the first derivative of PLR tracing), and the sustained constriction amplitude. For ERG measurements, both scotopic and photopic responses were recorded following stimulation with green light (520 nm) at preselected intensities. The amplitudes and latencies of the a-wave and the b-wave were also analyzed. In both strains, 1-month-old animals presented with a smaller baseline pupil diameter compared to that in 2- and 4-month-old mice. They also exhibited greater maximal constriction amplitude in response to red stimuli of medium intensity. Further, 1-month-old Sv129S6 mice responded with greater constriction amplitude to all other red and blue stimuli. One-month-old C57BL/6 mice also demonstrated faster early partial dilation and smaller sustained response to low blue stimuli. The ERG of 1-month-old C57BL/6 mice showed a greater scotopic a-wave amplitude compared to that of 2-month-old mice, whereas no significant differences were found in Sv129S6 mice. These results suggest that the functional maturation of the neuronal pathway that mediates the PLR continues after 1 month of age. In studies that measure PLR to determine retinal integrity in adult mice, it is thus important to determine normative values in animals of 2 months of age

A rapid and reliable determination of doxycycline hyclate by HPLC with UV detection in pharmaceutical samples

An accurate, sensitive and reproducible high performance liquid chromatographic (HPLC) method for the quantification of doxycycline hyclate in pharmaceutical samples has been developed and validated. The drug and the standard were eluted from a Lichrosorb RP-8 (250 mm´4.6 mm, 10 mm particle size) at 20 °C with a mobile phase consisting of methanol, acetonitrile and 0.010 M aqueous solution of oxalic acid (2:3:5, v/v/v). The flow rate was 1.25 ml min-1. A UV detector set at 350 nm was used to monitor the effluent. Each analysis required no longer than 4 min. The limits of detection and quantification were 1.15 and 3.84 μg ml-1, respectively. Recoveries for different concentrations ranged from 99.58 to 101.93 %