Preanalysis in Serum Hepcidin Measurement.

BACKGROUND: Hepcidin is a 25-amino peptide hormone that regulates iron homeostasis. Its serum quantification helps to provide the right therapeutic choice in iron-deficiency anemia and anemia in chronic diseases. Diurnal levels of serum iron might affect hepcidin secretion during the day. Blood collection time is an important part of the preanalytical phase of its quantification. METHODS: During the period 2013 - 2014, we collected blood samples for serum hepcidin quantification in 100 healthy controls. The samples were collected in vacuettes with serum separator gel at three different times during the day: 07:30 - 08:30, 12:00 - 13:00, and 16:00 - 17:00 hours. Hepcidin levels were measured with an ELISA method. RESULTS: We found a significant difference in serum hepcidin levels during the chosen three blood taking times. The normal range for Bulgarian population is 3.05 µg/L - 37.75 µg/L. The measured levels were: at 07:30 - 08:30 hours 12.2 µg/L (5.5 µg/L - 23.6 µg/L), 12:00 - 13:00 hours 14.1 µg/L (7.1 µg/L - 27.2 µg/L), and 16:00 - 17:00 hours 16.5 µtg/L (9.9 µg/L - 29.6 µpg/L) 10.7 < r < 1.0; p < 0.5 between 07:30 - 08:30 and 12:00 - 13:00 hours and p < 0.05 between 07:30 - 08:30 hours and 16:00 - 17:00 hours and 12:00 - 13:00 hours and 16:00 - 17:00 hours]. No significant differences were found for transferrin saturation between measured groups [0.1 < r < 0.3; p > 0.5]. CONCLUSIONS: In order to obtain the most correct results for serum hepcidin quantification (especially in border to referent range levels) in the preanalytical phase, it is important to consider the time of blood sampling.

First Report of Alder Yellows Phytoplasma Associated with Common Alder (Alnus glutinosa) in the Republic of Macedonia.

Alder yellows phytoplasma (AldYp) is classified as a member of the 16SrV-group of phytoplasmas and is closely related to Flavescence dorée (FD), a quarantined pathogen of economic importance affecting vineyards across Europe. AldYp is associated with common (Alnus glutinosa) and grey alder (A. incana), and has been reported in France, Italy, Germany, Austria, Switzerland, the Baltic region, Serbia, and Montenegro (1,2,4). For Macedonian vineyards, so far, neither infection of grapevine with 16SrV-group of phytoplasmas nor the presence of the main FD phytoplasma vector, Scaphoideus titanus, has been recorded. However, the presence of FD-related phytoplasma was detected in wild Clematis vitalba. In September and October 2013, leaves with petioles from A. glutinosa exhibiting leaf discoloration and yellowing were collected from two sites (41°23'43″ N, 22°54' E and 41°23' N, 22°53' E) in southeast Macedonia near the village of Smolare (Strumica district). Eight samples were collected from each site. Leaves of six asymptomatic alder seedlings collected from the same sites served as a control. Nucleic acids were extracted from fresh leaf midribs and petioles using a DNeasy Plant Mini Kit (Qiagen, Hilden, Germany). Initial phytoplasma identification was carried out by nested PCR assay of the 16S rRNA gene, using universal primers P1/P7 and R16F2n/R16R2 followed by RFLP with MseI endonuclease (Fermentas, Vilnius, Lithuania), as previously reported (4). Characterization of detected phytoplasmas was performed by amplifying two genetic loci specific for the members of the 16SrV group phytoplasmas; the ribosomal protein gene operon (rp) using primers rp(V)F1/rpR1 and rp(V)F1A/rp(V)R1A (3), and the non-ribosomal metionine aminopeptidase (map) gene using primer set FD9f5/MAPr1 and FD9f6/MAPr2 (1). The PCR amplicons were sequenced and deposited in NCBI GenBank database under the accession numbers KJ605448 to 52 (map) and KJ605453 to 57 (rp). The obtained sequences were compared with reference sequences of the 16SrV-group phytoplasmas (1,3) using the neighbor-joining method in MEGA5 (5). The presence of phytoplasma was detected in 14 of 16 symptomatic alder samples, while all control plants tested negative. The MseI restriction profiles were identical among all 14 samples and with the reference strains of the 16SrV group phytoplasmas (EY1 - 16SrV-A, FD-C - 16SrV-C, and FD-D - 16SrV-D). The rp-based phylogeny enabled identification of four diverse phytoplasma strains among the AldYp strains from Macedonia. Three strains clustered within the rpV-E subgroup while one belonged to rpV-L subgroup. Phylogenetic analysis of the more variable genetic locus, map, showed the presence of five diverse phytoplasma strains. Four strains belonged to the map-FD2 (FD-D, FD92) cluster, while one grouped within the map-FD1 (FD70) cluster. To our knowledge, this is the first report of 16SrV phytoplasma group occurrence on alder in Macedonia. The significant similarity between AldYp strains and FD sensu stricto indicate the risk of pathogen exchange between the wild ecosystem and the grapevine (1). Alder trees naturally infected with the FDp-related strains could therefore represent a serious risk for FD outbreak in Macedonian vineyards if local S. titanus populations developed. References: (1) G. Arnaud et al. Appl. Environ. Microbiol. 73:4001, 2007. (2) T. Cvrkovic et al. Plant Pathol. 57:773, 2008. (3) M. Martini et al. Int. J. Syst. Evol. Microbiol. 57:2037, 2007. (4) S. Radonjic et al. Plant Dis. 97:686, 2013. (5) K. Tamura et al. Mol. Biol. Evol. 28:2731, 2011.

Interactions of odorants with olfactory receptors and receptor neurons match the perceptual dynamics observed for woody and fruity odorant mixtures.

The present study aimed to create a direct bridge between observations on peripheral and central responses to odorant mixtures and their components. Three experiments were performed using mixtures of fruity (isoamyl acetate; ISO) and woody (whiskey lactone; WL) odorants known to contribute to some of the major notes in Burgundy red wine. These experiments consisted of (i) calcium imaging of human embryonic kidney cells (HEK293T) transfected with olfactory receptors (ORs); (ii) single-unit electrophysiological recordings from olfactory receptor neurons (ORNs) and analyses of electro-olfactogram (EOG) responses in the rat nose in vivo; and (iii) psychophysical measurements of the perceived intensity of the mixtures as rated by human subjects. The calcium imaging and electrophysiological results revealed that ISO and WL can act simultaneously on single ORs or ORNs and confirm that receptor responses to mixtures are not the result of a simple sum of the effects of the individual mixture compounds. The addition of WL to ISO principally suppressed the ORN activation induced by ISO alone and was found to enhance this activation in a subset of cases. In the human studies, the addition of high concentrations of WL to ISO decreased the perceived intensity of the ISO. In contrast, the addition of low concentrations of WL enhanced the perceived intensity of the fruity note (ISO) in this mixture, as it enhanced EOG responses in ORNs. Thus, both OR and ORN responses to ISO + WL mixtures faithfully reflected perceptual response changes, so the odour mixture information is set up after the peripheral stage of the olfactory system.

[The copper concentration in the blood serum of women with ovarian tumors (a preliminary report)]. / Kontsentratsiia na medta v kruvniia serum pri zheni s ovarialni tumori (predvaritelno suobshtenie).

The aim of the present study was to evaluate serum copper concentration changes in patients with benign cysts or carcinomas of ovary, and to compare these concentrations with the normal reference serum copper levels. Patients included in the study were as follows: benign ovarian cyst cases (n = 19), endometrial cyst cases (n = 5), ovarian carcinoma cases (n = 6), female patients with no ovarian disease (n = 161). Reference serum copper levels in apparently healthy subjects were 16.8 +/- 4.15 mumol/l (mean +/- SD). Serum copper concentration were significantly increased in ovarian carcinoma patients (22.98 +/- 3.90 mumol/l) in comparison with benign cyst patients (18.40 +/- 4.61 mumol/l) (p < 0.05), in with endometrial cyst patients (15.94 +/- 4.24 mumol/l) (p = 0.02), and in comparison with normal levels in controls (p < 0.001). Copper/zinc ratios were highest in ovarian carcinoma patients--1.68, intermediate in benign cyst patients--1.42, and lowest in endometrial cyst patients--1.08. The mechanisms underlying serum copper concentration variations in the different groups are briefly discussed with special emphasis on these in ovarian carcinoma patients.

[Tocolytic treatment of pregnant women with threatened abortions and preterm labor in the second and third trimesters of pregnancy with magne B6 (a preliminary communication)]. / Tokolitichno lechenie pri bremenni sus zaplashvashti aborti i prezhdevremenni razhdaniia vuv vtoriia i tretiia trimestur na bremennostta s preparat "Magne B6" (predvaritelno suobshtenie).

The authors check the effect of the substitutional treatment of the pregnant patients with the uterine paints and contractions in the second and third months periods of the pregnancy with "Magne B6"--Sanofi. In the group of the examined patients the Mg deficiency that is the cause for these suffering has been established. After the substitutional treatment with "Magne B6" the described suffering disappeared and clinical-lab values of Mg in the serum and urine normalized.

[Multiple pregnancy and microelements: zinc (preliminary report)]. / Mnogoplodna bremennost i mikroelementi: tsink (predvaritelno suobshtenie).

The aim af the study was to define the concentration of zinc in sera of women with multiple pregnancies during the last trimester and to compare it with the values in normal pregnancies. The study included 12 women with gemini and 67 with single fetus pregnancies. A statistically significant decrease was found in the mean zinc concentration in twin pregnancies--8.1 +/- 1.72 mcmol/L compared to the mean values in normal pregnancies--10.5 +/- 1.90 mcmol/L (p < 0.001). Various mechanisms, possibly responsible for the stated changes, their relation to some complications of multiple pregnancy and delivery are discussed, as well as their prophylactics.

[The zinc concentration of the blood serum in women with ovarian tumors (preliminary report)]. / Kontsentratsiia na tsinka v kruvniia serum pri zheni s tumori na iaichnitsite (predvaritelno suobshtenie).

The aim of the study is to determine the concentration of blood serum zinc of women with ovarian tumors. The patients included in the study are as follows: 15 women with benign ovarian cysts, 5 with endometrial cysts and 5 with ovarian carcinoma. The control group consists of 157 healthy women whose serum zinc concentration was 13.50 +/- 2.60 mmol/L. We could not demonstrate a significant difference in serum zinc concentration of different groups: benign cysts, endometrial cysts and ovarian carcinoma in comparison control group.