Association of LOXL1 gene common sequence variants in Jordanian patients with exfoliation syndrome and exfoliative glaucoma

AIM: To investigate the association between single nucleotide polymorphisms (SNPs) in the LOXL1 gene with exfoliation syndrome/glaucoma (XFS/XFG) among Jordanians. METHODS: Sixty-one patients with XFS/XFG and 59 healthy control individuals were recruited in the study. Patients were diagnosed with XFS/XFG using standard clinical examination techniques. The exonic rs1048661 SNP and the intronic rs2165241 SNP in LOXL1 gene were genotyped using sequencing technique. Allele and genotype frequencies were compared between cases and controls using Chi-square analysis. RESULTS: The G allele of the rs1048661 SNP and the T allele of the rs2165241 SNP were common in the sample with frequencies of 86.4% and 81.4%, respectively. In addition, there were no significant differences in the genotypic and allelic distributions between patients and controls for rs1048661 SNP (P=0.770, OR=1.21, 95%CI: 0.56-2.60) and for rs2165241 SNP (P=0.605, OR=1.12, 95%CI: 0.59-2.09). In addition, no significant associations were found between haplotypes of the examined SNPs and XFS/XFG in the sample (P>0.05). CONCLUSION: Variations in LOXL1 gene may not be associated with XFS/XFG in the Jordanian population. More studies are required to confirm the current findings

Procathepsin V Is Secreted in a TSH Regulated Manner from Human Thyroid Epithelial Cells and Is Accessible to an Activity-Based Probe

The significance of cysteine cathepsins for the liberation of thyroid hormones from the precursor thyroglobulin was previously shown by in vivo and in vitro studies. Cathepsin L is most important for thyroglobulin processing in mice. The present study aims at specifying the possible contribution of its closest relative, cysteine cathepsin L2/V, to thyroid function. Immunofluorescence analysis on normal human thyroid tissue revealed its predominant localization at the apical plasma membrane of thyrocytes and within the follicle lumen, indicating the secretion of cathepsin V and extracellular tasks rather than its acting within endo-lysosomes. To explore the trafficking pathways of cathepsin V in more detail, a chimeric protein consisting of human cathepsin V tagged with green fluorescent protein (GFP) was stably expressed in the Nthy-ori 3-1 thyroid epithelial cell line. Colocalization studies with compartment-specific markers and analyses of post-translational modifications revealed that the chimeric protein was sorted into the lumen of the endoplasmic reticulum and subsequently transported to the Golgi apparatus, while being N-glycosylated. Immunoblotting showed that the chimeric protein reached endo-lysosomes and it became secreted from the transduced cells. Astonishingly, thyroid stimulating hormone (TSH)-induced secretion of GFP-tagged cathepsin V occurred as the proform, suggesting that TSH upregulates its transport to the plasma membrane before it reaches endo-lysosomes for maturation. The proform of cathepsin V was found to be reactive with the activity-based probe DCG-04, suggesting that it possesses catalytic activity. We propose that TSH-stimulated secretion of procathepsin V is the default pathway in the thyroid to enable its contribution to thyroglobulin processing by extracellular means

Development of a Millet Starch Edible Film Containing Clove Essential Oil

Medicinal plants contain various secondary metabolites. The present study analyzed the essential oil of buds from clove (Syzygium aromaticum L.; Family: Myrtaceae) using gas chromatography-mass spectrometry (GC-MS). GC-MS analysis showed the presence of six major phytoconstituents, such as eugenol (66.01%), caryophyllene (19.88%), caryophyllene oxide (5.80%), phenol, 2-methoxy-4-(2-propenyl)-acetate (4.55%), and humulene (3.75%). The effect of clove essential oils (CEO) at 0%, 1%, 2%, and 3% (w/w) on the mechanical and barrier properties of starch films was evaluated. The tensile strength (TS) and elongation (E) of films with clove essential oil were 6.25 ± 0.03 MPa and 5.67% ± 0.08%, respectively. The antioxidant activity of the films significantly increased the millet starch film and presented the lowest antioxidant activity (0.3%) at a 30 minute incubation for the control sample, while increasing CEO fraction in the starch film lead to an increase in antioxidant activity, and the 3% CEO combined film presented the highest antioxidant activity (15.96%) at 90 min incubation. This finding could be explained by the incorporation of clove oil containing antioxidant properties that significantly increased with the incorporation of CEO (p < 0.05). A zone of inhibition ranging from 16 to 27 mm in diameter was obtained when using a concentration of CEO ranging from 1% to 3%. We also observed the presence of an antimicrobial activity on several tested microorganism including Escherichia coli, Pseudomonas aeruginosa, Enterobacter sp, Bacillus cereus, Staphylococcus aureus, and Trichoderma fungi. Thus, the current study reveals the possibility of using a millet starch edible film as a preservation method

Serum Vitamin D level in Jordanian patients with exfoliation syndrome and exfoliative glaucoma

AIM: To compare the level of serum Vitamin D in patients with exfoliation syndrome(XFS)and exfoliative glaucoma(XFG)with that in control subjects.METHODS: Serum levels of 25-hydroxy Vitamin D(Vitamin D)were measured using liquid chromatography. Variables like age, sex and medical history of hypertension, diabetes mellitus and ischemic heart disease were reported.RESULTS: This study included a total of 55 patients with XFS/XFG and 60 control subjects. Patients with XFS/XFG were significantly older than control subjects(mean age: 71.8y vs 67.5y, P=0.002). Gender distribution was similar. The prevalence rates of diabetes, hypertension, and ischemic heart disease were not significantly different between the two groups. The mean of Vitamin D was 14.7 ng/mL for patients with XFS/XFG and 14.9 ng/mL for control subjects with no significant difference. Although Vitamin D level was lower in the XFS/XFG group but multivariate analysis did not show significant difference in Vitamin D deficiency between the two groups after adjusting for age, gender, and medical conditions.CONCLUSION: The outcomes of our study are different from those in the literature. Vitamin D levels were low in both the XFS/XFG group and the control group being lower in the first one but that difference was not statistically significant. XFS and Vitamin D deficiency share common associations with certain systemic diseases. Further studies with larger numbers are needed to elaborate more on these associations and to support further the controversial literature

Treatment of rat thyrocytes in vitro with cathepsin B and L inhibitors results in disruption of primary cilia leading to redistribution of the trace amine associated receptor 1 to the endoplasmic reticulum

Taar1 is a G protein-coupled receptor (GPCR) confined to primary cilia of rodent thyroid epithelial cells. Taar1-deficient mouse thyroid follicles feature luminal accumulation of thyroglobulin suggesting that Taar1 acts as a regulator of extra- and pericellular thyroglobulin processing, which is mediated by cysteine cathepsin proteases present at the apical plasma membrane of rodent thyrocytes. Here, by immunostaining and confocal laser scanning microscopy, we demonstrated co-localization of cathepsin L, but only little cathepsin B, with Taar1 at primary cilia of rat thyrocytes, the FRT cells. Because proteases were shown to affect half-lives of certain receptors, we determined the effect of cathepsin activity inhibition on sub-cellular localization of Taar1 in FRT cells, whereupon Taar1 localization altered such that it was retained in compartments of the secretory pathway. Since the same effect on Taar1 localization was observed in both cathepsin B and L inhibitor-treated cells, the interaction of cathepsin activities and sub-cellular localization of Taar1 was thought to be indirect. Indeed, we observed that cathepsin inhibition resulted in a lack of primary cilia from FRT cells. Next, we proved that primary cilia are a necessity for Taar1 trafficking to reach the plasma membrane of FRT cells, since the disruption of primary cilia by treatment with β-cyclodextrin resulted in Taar1 retention in compartments of the secretory pathway. Furthermore, in less well-polarized rat thyrocytes, namely in FRTL-5 cells lacking primary cilia, Taar1 was mainly confined to the compartments of the secretory pathway. We conclude that Taar1 localization in polarized thyroid epithelial cells requires the presence of primary cilia, which is dependent on the proteolytic activity of cysteine cathepsins B and L

La Croix du Nord : supplément régional à la Croix de Paris ["puis" grand journal quotidien du Nord de la France]

14 mai 19121912/05/14 (A23,N7864).Appartient à l’ensemble documentaire : NordPdeC

Hydrophobic natural deep eutectic-solvent bar microextraction combined with HPLC-DAD for determination of urinary 1-hydroxypyrene of smokers

1-hydroxypyrene (1-OHP) is a metabolite of pyrene, which is a type of polycyclic aromatic hydrocarbons (PAHs) found in tobacco smoke and preferred use as a biomarker to determine exposure to PAHs from smoking. This paper reports a new and simple microextraction technique for 1-hydroxypyrene (1-OHP) determination in smokers' urines using hydrophobic natural deep eutectic solvent (HNDES) in developing solvent bar microextraction (SBME) and high performance liquid chromatography-diode array detection (HPLC-DAD). Terpenoids based-HNDES using various ratios of (thymol/menthol) were synthesized and investigated as extraction solvents. The variables affecting HNDE-SBME steps were optimized by response surface methodology (RSM) based on central composite design (CCD). Under the optimum conditions, the calibration graph of spiked urine samples was linear in the range of 0.3–25.0 μg L−1 1-OHP with a correlation coefficient of 0.999. Additionally, the detection limit, quantitation limit, and extraction recovery were found to be 0.06 μg L−1, 0.22 μg L−1, and ≥100%, respectively. In comparison with referenced methods and based on the satisfactory results of applications, the proposed method can be considered as a potential method for the detection of urinary 1-OHP for health risk assessment of PAHs exposures

Additional file 3: Figure S3. of Assessment of cytotoxicity exerted by leaf extracts from plants of the genus Rhododendron towards epidermal keratinocytes and intestine epithelial cells

Overview of mitochondrial morphology in IEC6 and HaCaT cells after a 24h-exposure to three different concentrations (5, 50 and 500 μg/mL) of Rhododendron leaf extracts. Confocal fluorescence images of IEC6 (a, left) and HaCaT (b, right) cells labeled with MitoTracker® Red CMXRos. Cells treated with 0.5 % DMSO served as controls, A) R. hippophaeoides var. hippophaeoides, B) R. minus, C) R. rubiginosum, D) R. cinnabarinum, E) R. ferrugineum, F) R. polycladum, G) R. concinnum, H) R. xanthostephanum, I) R. anthopogon ssp. anthopogon, J) R. ambiguum, K) R. hirsutum, and L) R. racemosum. Bars represent 50 μm. (TIFF 11640 kb