This study investigated quantitated expression of dopamine 2 receptor (D2R) and somatostatin receptors of the five types (SSTR1-SSTR5) in a large series of clinically non-functioning pituitary adenomas (CNFAs). Co-expression of these receptors in individual adenomas was studied as well as correlation between receptor types. Adenoma tissue from 198 patients who underwent surgery for CNFAs was analyzed by immunohistochemistry and quantitative real-time PCR. D2R and SSTR1-3 mRNA was expressed in all 198 adenomas. SSTR4 and SSTR5 were detectable in 85 % and 61 % of adenomas, respectively. Expression of D2R was significantly higher than that of the somatostatin receptors. The median relative expressions were as follows from highest D2R >> SSTR3 > SSTR2 > SSTR1 > SSTR5 > SSTR4. High relative expression (ratio to β-glucuronidase mRNA > 1) of D2R was found in 60 % of tumors, high expression of SSTR1 in 7.5 %, SSTR2 in 7 %, SSTR3 in 4 % and SSTR5 in 0.5 %. The quantity of D2R correlated positively with expression of SSTR2 and SSTR3, and negatively with SSTR1 and SSTR5. Among histological adenoma types, SSTR1 was significantly higher in null-cell adenomas and SSTR3 was lower in silent corticotroph adenomas. In conclusions, in CNFAs, high expression of somatostatin receptors is much less common than that of D2R, and co-expression of both these receptors is exceptional. D2R and SSTR3 seem to be the most promising targets for pharmacological treatment., F. Gabalec, M. Drastikova, T. Cesak, D. Netuka, V. Masopust, J. Machac, J. marek, J. Cap, M. Beranek., and Obsahuje bibliografii
Saliva-activated transmission (SAT) of Borrelia burgdorferi sensu stricto was demonstrated using real-time PCR and salivary gland extract (SGE) from partially fed Ixodes ricinus ticks. C3H/HeN mice were injected intradermally with 1.5 × 103 spirochetes mixed with 40 µg of SGE per mouse. The control group was inoculated with the same dose of spirochetes without SGE. The accelerating effect of SGE on spirochete proliferation was demonstrated on day 1 post infection, when a 4.2-fold increase in spirochetes was found in the skin and a 10-fold increase in the blood, compared with control mice. The data represent the first direct evidence of a SAT effect of I. ricinus SGE on infection with the Lyme disease agent B. burgdorferi.
Due to inconsistent results of APOE variants in the survival of pregnancy we investigated the potential relationship of APOE rs7412 and rs429358 with pregnancy loss (PL) in Bosnian women. We enrolled 154 women with PL. The minimum week of miscarriage was 6, while the maximum was 28. As a control group, an equal number of mothers with at least one live-born child was included. All women were recruited from the Institution of Health Protection of Women and Motherhood in Sarajevo, Bosnia and Herzegovina. Genotyping was performed by re-al-time PCR at the Department of General Pha-macology and Pharmacoeconomics, Pomeranian Medical University. The prevalence of genotypes E2/E3, E2/E4, E3/E3, E3/E4, E4/E4 in the group with and without PL were: 14.3 %, 1.3 %, 70.8 %, 12.3 %, 1.3 %, and 13.6 %, 1.3 %, 70.1 %, 14.3 %, 0.7 %, respectively. The frequency of the E4/E4 genotype in women with 1–2 and 3–4 PL compared to women without PL did not differ significantly between those three groups (P value = 0.0712). The frequencies of alleles ԑ2, ԑ3, ԑ4 in the group with and without PL, were: 6.8 %, 85.1 %, 8.1 % and 7.5 %, 84.1 %, 8.4 %, respectively, and did not differ significantly. We conclude that our study does not confirm rs7412 and rs429358 as a potential risk factor for PL in the studied group. To elucidate the relationship between PL and variants of the APOE gene, studies with a larger sample size and placental histomorphology and ge-netic diagnosis are required. and Corresponding author: Grażyna Adler
Response mechanisms of rainbow trout Oncorhynchus mykiss (Walbaum), experimentally infected with a Danish strain of Gyrodactylus salaris Malmberg, 1957 were investigated using molecular tools (qPCR) and immunohistochemistry. Expression of ten immune-relevant genes and reactivity with five different antibodies in the epidermis of skin and fin tissue were analysed in susceptible but responding rainbow trout. Rainbow trout were susceptible with regard to the parasite strain which initially colonised fins but relocated to the body region as infection progressed. The ten investigated genes encoding the cytokines IL-1β, TNF-α, IFN-γ, IL-10 and markers for adaptive immune activity, such as CD-4, CD-8, TCR-α, IgM, IgT and MHC II, were not found significantly regulated during the course of infection although IFN-γ showed a slight up-regulation. Immunohistochemical analyses showed positive reactivity with antibodies against CD3, B-lymphocytes, neutrophilic granulocytes and collectin but not with mAb against IgM. No staining differences between infected and non-infected skin and fin tissue were detected.
Despite the global distribution of the brown dog tick, Rhipicephalus sanguineus (Latreille, 1806) sensu lato (s.l.), limited information exists about their identity from the Arabian Peninsula. Ticks from free roaming urban dogs and dromedary camels in Riyadh, Saudi Arabia were morphologically identified, confirmed with scanning electron microscopy and characterised at mitochondrial DNA (cox1, 12S rDNA and 16S rDNA). A total of 186 ticks were collected from 65 free roaming dogs (n = 73) and 84 dromedary camels (n = 113). Morphologically, 5.9% (11/186) were R. sanguineus s.l. and Hyalomma spp. (93.5%, 174/186). From within R. sanguineus s.l., the presence of Rhipicephalus cf. camicasi Morel, Mouchet et Rodhain, 1976 (1 dog, 2 camels) and Rhipicephalus turanicus Pomerantsev, 1936 (1 camel) is reported. The examined R. cf. camicasi form a sister group to R. sanguineus s.l. tropical lineage at all DNA markers. Dogs were parasitised by Hyalomma dromedarii Koch, 1844 (n = 59), Hyalomma impeltatum Schulze et Schlottke, 1930 (n = 1), Hyalomma excavatum Koch, 1844 (n = 2), Hyalomma turanicum Pomerantsev, 1946 (n = 1) and Hyalomma rufipes Koch,1844 (n = 1). DNA from dog blood (n = 53) from Riyadh confirmed a low prevalence of canine vector-borne pathogens that does not exceed 5.7% for Babesia spp., Mycoplasma spp., Anaplasma platys, Hepatozoon canis and Ehrlichia canis using multiplexed tandem PCR (MT-PCR) and diagnostic PCR. Low prevalence of R. sanguineus s.l. on dogs likely contributed to the low level of canine vector-borne pathogens in Saudi Arabia. We demonstrate that dogs in the central Arabian Peninsula are more commonly parasitised by Hyalomma spp. than R. sanguineus s.l., Shona Chandra, Karen Smith, Abdullah D. Alanazi, Mohamed S. Alyousif, David Emery, Jan Šlapeta., and Obsahuje bibliografii
The mitochondrial DNA (mtDNA) amount in cells as the basis for mitochondrial energy generating system, which produces ATP, plays an important role in the fetal development and postnatal morbidity. Isolated human cord blood leukocytes (HCBL) contribute very little to the overall metabolic turnover, but they may serve as easily available marker cells for the study of the mtDNA amount changes in cord blood during fetal development. The aim of our study was to analyze the mtDNA amount in HCBL. HCBL were isolated from cord blood samples of 107 neonates born between the 25th and 41st week of gestation. The mtDNA amount was analyzed by the real-time PCR method. The significant negative correlations were found between the relative mtDNA amount in HCBL and gestational age (r = -0.54, p<0.01) and birth weight (r = -0.43, p<0.01), respectively. The results revealed that the mtDNA content per cell decreases in HCBL with progressing fetal development. This may be explained by gradual shift of the hematopoiesis from fetal liver to bone marrow during the second half of pregnancy presumably accompanied by decreasing cell volume of HCBL as it was shown similarly in red blood cells., M. Pejznochová, M. Tesařová, T. Honzík, H. Hansíková, M. Magner, J. Zeman., and Obsahuje bibliografii a bibliografické odkazy
High incidence of thrombosis and venous thromboembolism was reported in patients with COVID-19. In this study, we focused on analysis of thrombophilic mutations performed without a standard DNA extraction step. In one hundred of COVID-19 positive outpatients, real-time PCR for Leiden mutation in the FV gene and G20210A mutation in the FII gene was carried out from DNA extracts and modified whole blood samples, and their cycle threshold (Ct) values were evaluated. In the extracts, healthy homozygotes (wt/wt), heterozygotes (M/wt), and homozygous carriers of Leiden mutation (M/M) provided median Ct values of 18.5, 19.4/22.0, and 20.9. In the whole blood, Ct values were 25.3 (wt/wt), 24.8/27.2 (M/wt), and 26.9 (M/M). Median Ct values for G20210A in the extracts were 19.6 for homozygotes (wt/wt), and 19.7/20.4 for heterozygous carriers. The whole blood samples provided Ct values of 23.9 in healthy homozygotes and 26.3/27.2 in heterozygotes for G20210A mutation. No homozygous subjects for G20210A and no double heterozygotes (for Leiden and G20210A mutations) were found. Despite significant differences in the Ct values, genotyping showed complete result concordance of the DNA extracts and the whole blood samples. The integrity and amplificability of DNA molecules in the whole blood samples during 28 days of deep freezing, interrupted by four cycles of thawing, did not significantly change. In conclusion, we demonstrated a new protocol for the detection of the thrombophilic mutations via real-time PCR on the modified whole blood of COVID-19 positive patients. The blood modification was reliable, easy, cheap, and saving costs and turnaround time of the whole laboratory process.