Clinical adaptation of a high-performance liquid chromatographic method for the assay of pyridoxal 5′-phosphate in human plasma

Vitamin B6, measured as pyridoxal 5′-phosphate (PLP), is a co-enzyme in the transsulfuration pathway of homocysteine metabolism. Since depletion of PLP has been suggested as an independent risk factor for coronary artery disease, PLP is frequently measured to guide patient care. By a change and utilization of an Aquasil C18 column and the addition of an acetonitrile clean-up gradient to the potassium phosphate, with sodium perchlorate and bisulfite buffer between samples we report the modification of a previously described method for analysis of PLP. The result is a more practical, efficient, reliable and robust method for daily clinical use. We also determined and report that it is critical to protect freshly prepared standard PLP samples from light exposure during assay preparation.     

Cu(II)—(lAsp)n system. Spectroscopic evidence of hexatomic chelate ring formation

The study of the Cu(II)-(L Asp)_n system using circular dichroism and potentiometric data has provided evidence indicating the formation of two complexes in a two step process. In the first (I) of these complexes, obtained at pH 4.5, two carboxyl residues are bound to the metal. This complex partially inhibits the transition from α helix to nonperiodic conformation. In the second complex (II) two peptide nitrogens and two carboxylate oxygens are bound to each Cu(II) ion forming two hexatomic chelate rings. The CD spectral pattern is then the opposite of what is obtained when a five-membered chelate ring is formed.     

Localization of the Brainstem GABAergic Neurons Controlling Paradoxical (REM) Sleep

Paradoxical sleep (PS) is a state characterized by cortical activation, rapid eye movements and muscle atonia. Fifty years after its discovery, the neuronal network responsible for the genesis of PS has been only partially identified. We recently proposed that GABAergic neurons would have a pivotal role in that network. To localize these GABAergic neurons, we combined immunohistochemical detection of Fos with non-radioactive in situ hybridization of GAD67 mRNA (GABA synthesis enzyme) in control rats, rats deprived of PS for 72 h and rats allowed to recover after such deprivation. Here we show that GABAergic neurons gating PS (PS-off neurons) are principally located in the ventrolateral periaqueductal gray (vlPAG) and the dorsal part of the deep mesencephalic reticular nucleus immediately ventral to it (dDpMe). Furthermore, iontophoretic application of muscimol for 20 min in this area in head-restrained rats induced a strong and significant increase in PS quantities compared to saline. In addition, we found a large number of GABAergic PS-on neurons in the vlPAG/dDPMe region and the medullary reticular nuclei known to generate muscle atonia during PS. Finally, we showed that PS-on neurons triggering PS localized in the SLD are not GABAergic. Altogether, our results indicate that multiple populations of PS-on GABAergic neurons are distributed in the brainstem while only one population of PS-off GABAergic neurons localized in the vlPAG/dDpMe region exist. From these results, we propose a revised model for PS control in which GABAergic PS-on and PS-off neurons localized in the vlPAG/dDPMe region play leading roles.     

Increased Retinal Ganglion Cell Survival by Exogenous IL-2 Depends on IL-10, Dopamine D1 Receptors,and Classical IL-2/IL-2R Signaling Pathways

Neurochemical Research - Interleukin-2 (IL-2) is a classical pro-inflammatory cytokine known to display neuroprotective roles in the central nervous system including the retina. In the present...     

Combined arene ruthenium porphyrins as chemotherapeutics and photosensitizers for cancer therapy

Mononuclear 5-(4-pyridyl)-10,15,20-triphenylporphyrin and 5-(3-pyridyl)-10,15,20-triphenylporphyrin as well as tetranuclear 5,10,15,20-tetra(4-pyridyl)porphyrin (tetra-4-pp) and 5,10,15,20-tetra(3-pyridyl)porphyrin) (tetra-3-pp) arene ruthenium(II) derivatives (arene is C₆H₅Me or p-Pr ⁱ C₆H₄Me) were prepared and evaluated as potential dual photosensitizers and chemotherapeutics in human Me300 melanoma cells. In the absence of light, all tetranuclear complexes were cytotoxic (IC₅₀ ≤ 20 μM), while the mononuclear derivatives were not (IC₅₀ ≥ 100 μM). Kinetic studies of tritiated thymidine and tritiated leucine incorporations in cells exposed to a low concentration (5 μM) of tetranuclear p-cymene derivatives demonstrated a rapid inhibition of DNA synthesis, while protein synthesis was inhibited only later, suggesting arene ruthenium–DNA interactions as the initial cytotoxic process. All complexes exhibited phototoxicities toward melanoma cells when exposed to laser light of 652 nm. At low concentration (5 μM), LD₅₀ of the mononuclear derivatives was between 5 and 10 J/cm², while for the tetranuclear derivatives LD₅₀ was approximately 2.5 J/cm² for the [Ru₄(η⁶-arene)₄(tetra-4-pp)Cl₈] complexes and less than 0.5 J/cm² for the [Ru₄(η⁶-arene)₄(tetra-3-pp)Cl₈] complexes. Examination of cells under a fluorescence microscope revealed the [Ru₄(η⁶-arene)₄(tetra-4-pp)Cl₈] complexes as cytoplasmic aggregates, whereas the [Ru₄(η⁶-arene)₄(tetra-3-pp)Cl₈] complexes were homogenously dispersed in the cytoplasm. Thus, these complexes present a dual synergistic effect with good properties of both the arene ruthenium chemotherapeutics and the porphyrin photosensitizer.     

Therapeutic Use of a Selective S1P1 Receptor Modulator Ponesimod in Autoimmune Diabetes

In the present study, we investigated the therapeutic potential of a selective S1P₁ receptor modulator, ponesimod, to protect and reverse autoimmune diabetes in non-obese diabetic (NOD) mice. Ponesimod was administered orally to NOD mice starting at 6, 10, 13 and 16 weeks of age up to 35 weeks of age or to NOD mice showing recent onset diabetes. Peripheral blood and spleen B and T cell counts were significantly reduced after ponesimod administration. In pancreatic lymph nodes, B lymphocytes were increased and expressed a transitional 1-like phenotype. Chronic oral ponesimod treatment efficiently prevented autoimmune diabetes in 6, 10 and 16 week-old pre-diabetic NOD mice. Treatment withdrawal led to synchronized disease relapse. Ponesimod did not inhibit the differentiation of autoreactive T cells as assessed by adoptive transfer of lymphocytes from treated disease-free NOD mice. In addition, it did not affect the migration, proliferation and activation of transgenic BDC2.5 cells into the target tissue. However, ponesimod inhibited spreading of the T cell responses to islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP). Treatment of diabetic NOD mice with ponesimod induced disease remission. However, here again, upon treatment cessation, the disease rapidly recurred. This recurrence was effectively prevented by combination treatment with a CD3 antibody leading to the restoration of self-tolerance. In conclusion, treatment with a selective S1P₁ modulator in combination with CD3 antibody represents a promising therapeutic approach for the treatment of autoimmune diabetes.     

Prevalence of Trachoma in the North Region of Cameroon: Results of a Survey in 15 Health Districts

Background

To estimate the prevalence of trachoma in the North Region of Cameroon in order to facilitate the planning of trachoma control activities in this region, a survey was carried out in 2011 and 2012 in 15 health districts (HDs).

Methodology

A cross-sectional, two-stage cluster random sampling survey was carried out. The survey focused on two target populations: children aged 1 to 9 years for the prevalence of Trachomatous Inflammation-Follicular (TF) and those aged 15 and over for the prevalence of Trachomatous Trichiasis (TT). The sample frame was an exhaustive list of villages and neighborhoods of HDs. The World Health Organization simplified trachoma grading system was used for the recognition and registration of cases of trachoma.

Principal Findings

30,562 children aged 1 to 9 years and 24,864 people aged 15 and above were examined. In children aged 1–9 years, the overall prevalence of TF was 4.2% (95% confidence intervals (CI): 4.0–4.5%). Three (3) of 15 HDs in the region showed TF prevalence of ≥10% (Poli, Rey Bouba, and Tcholliré). The overall TT prevalence was 0.25% (95% CI: 0.20–0.33%). There were estimated 1265 TT cases in the region. The prevalence of blindness was 0.01% (95% CI: 0.00–0.03%), low vision was 0.11% (95% CI: 0.07–0.17%), and corneal opacity was 0.22% (95% CI: 0.17–0.29%).

Conclusions/Significance

This survey provides baseline data for the planning of activities to control trachoma in the region. The overall prevalence of TF in the region is 4.2%, and that of TT is 0.2%; three HDs have a TF prevalence ≥10%. These three HDs are eligible for mass drug administration with azythromycin, along with the implementation of the “F” and “E” components of the SAFE strategy.     

The genus Opisthacanthus Peters, 1861 (Scorpiones: Hormuridae), a remarkable Gondwanian group of scorpions

New comments are proposed on the geographic distribution of genus Opisthacanthus, and the Gondwanian model is further supported. The diversity of the genus is extraordinary in Madagascar, with the same number of species as in continental Africa, but sub-Saharan Africa is home to six out of the nine groups currently recognized of Opisthacanthus. Given the affinities of the Opisthacanthus groups and their current distribution, a center of origin in Africa could be favored for these ancient scorpions. The proposed Gondwana model suggests that the Madagascar Opisthacanthus are closer to those of the New World, which is consistent with the affinities observed in morphological characters. A new species, Opisthacanthus titanus sp. n., is described from the Torotorofotsy Forest, located in Eastern Madagascar. The new species shows affinities with both Opisthacanthus madagascariensis Kraepelin, 1894 known from dry regions in the western portion of the island and Opisthacanthus lavasoa Lourenço, Wilmé & Waeber, 2016 only known from the extreme southeast of the island. The new species and O. madagascariensis have similar external morphologies but the morphometric values are markedly distinct. Moreover, O. madagascariensis is exclusively found in spiny forest thickets and open woodlands, whereas the new species was found in the humid forest of Torotorofotsy. The total number of species in Madagascar is now raised to twelve. Biogeographical scenarios are also proposed to infer the origin of the Opisthacanthus and better understand its distribution in the New World, in Africa and Madagascar.