Effect of Trichostatin A on CD4 surface density in peripheral blood T cells.

Acetylation level of chromatin histones is maintained by histone acetylases (HATs) and deacetylases (HDACs) and correlates with transcriptional activity of genes. Trichostatin A (TSA) is HDAC inhibitor that causes various effects in cells, including immunomodulation. The CD4 antigen is a key coreceptor involved in activation of T helper cells. Using quantitative real-time PCR (RQ-PCR) and flow cytometry techniques, we estimated CD4 transcript level and density of CD4 antigen on the surface of TSA-treated stimulated and unstimulated peripheral T cells. We observed a dose dependent decrease in CD4 mRNA level and antigen density on surface of TSA-treated stimulated T cells. However, we did not observe any significant TSA effect on CD4 mRNA and protein expression in unstimulated T cells. Our data suggest that TSA may induce biosynthesis of factors responsible for negative regulation of CD4 antigen expression in stimulated T cells. Our investigation may support previous observation that this drug has immunosuppresive effect on primary T cells and may be useful in treatment of certain autoimmune disorders

The role of DNA methylation in cancer development.

Epigenetic modifications include DNA methylation and covalent modification of histones. These alterations are reversible but very stable and exert a significant impact on the regulation of gene expression. Changes in methylation of promoter or first exon may mimic the effect of mutations of various tumor suppressor genes (TSGs) or protooncogenes. Carcinogenesis can also result from aberrations in genomic DNA methylation that include hypermethylation and hypomethylation of promoter or first exon of cancer-related genes. Hypermethylation of promoter of various TSGs causes their transcriptional silencing. However, hypomethylation of regulatory DNA sequences activates transcription of protooncogenes, retrotransposons, as well as genes encoding proteins involved in genomic instability and malignant cell metastasis. The methylation of genomic DNA in malignant cells is catalyzed by DNA methyltransferases DNMT1 and DNMT3B, revealing significantly elevated expression in different types of cancers. The reversibility of hypermethylation can be used as target of therapeutic treatment in cancer. DNMT 1 and DNMT3B inhibitors including 5-Aza-2'-deoxycytidine and antisense oligonucleotides have been applied in clinical trials of such treatment. Identification of aberrations of DNA methylation in cancer cells is a new field of investigation in carcinogenesis. We believe that epigenetic cancer diagnostic and therapy will be achieved in the next decades

Zaburzenia czynności kanałów jonowych w patogenezie padaczek idiopatycznych

Pomimo postępów diagnostyki nadal u ok. 60–65% chorych nie można jednoznacznie ustalić przyczyny padaczki. W tej grupie chorych główną rolę odgrywają przypuszczalnie czynniki genetyczne. Uważa się, że u ok. 40% pacjentów predyspozycja genetyczna odpowiada za występowanie padaczek, określanych mianem „idiopatycznych”. Podłoże genetyczne padaczki potwierdzają liczne przykłady rodzinnie występujących zespołów padaczkowych. Należą do nich dziedziczona autosomalnie dominująco padaczka czołowa z napadami nocnymi oraz młodzieńcza padaczka miokloniczna. Obydwie formy padaczek uwarunkowane są mutacjami genów dla podjednostek neuronalnego receptora nikotynowego dla acetylocholiny. Postęp genetyki stworzył szansę dokładniejszego zrozumienia epileptogenezy na poziomie molekularnym, co ułatwia rozpoznanie oraz stwarza bardziej racjonalne podstawy leczenia i zapobiegania tej postaci padaczki

Association between fertilin beta, protamines 1 and 2 and spermatid-specific linker histone H1-like protein mRNA levels, fertilization ability of human spermatozoa, and quality of preimplantation embryos.

Fertilization involves a series of cellular interactions culminating in the fusion of gamete membranes, creating a zygote and then an embryo. During the process of human fertilization in vivo or in conventional in vitro fertilization (IVF), sperm must be capable of undergoing the acrosome reaction, binding to the zona pellucida (ZP), and penetrating the ZP to fuse with the oolema. The key role in this process is played by fertilin beta. Protamines and histones are the proteins that bind to sperm chromatin and contribute in chromatin remodeling during early spermiogenesis. It has been suggested that these proteins may also participate in successful fertilization and embryo development. Using reverse transcription and real-time quantitative PCR reaction (QR-PCR) methods and zygote and embryo scoring, we compared fertilin beta, protamine 1 (PRM1), protamine 2 (PRM2), spermatid-specific linker histone 1 (HILS1) mRNAs levels, in vitro fertilization ability of mature spermatozoa, and quality of embryos obtained from in vitro fertilization (IVF). We found significantly lower contents of fertilin beta transcript in spermatozoa from patients in which IVF fertilization failed (

Monocyte Chemoattractant Protein-1− 2518 A/G Single Nucleotide Polymorphism Might Be Associated with Renal Disease and Thrombocytopenia of SLE

There is conflicting evidence on the contribution of the MCP-1 −2518 A>G (rs 1024611) polymorphism to SLE incidence and clinical manifestations. We examined the prevalence of the MCP-1 −2518 A>G polymorphism in SLE patients (n = 199) and controls (n = 250) in Poland. We did not observe a significant difference in the distribution of MCP-1 −2518 A>G polymorphic variants in patients with SLE and healthy individuals. However, we found an association between the GG versus AG and AA genotypes as well as the AG and GG versus AA genotypes with renal manifestations of SLE OR = 3.614 (1.123–11.631, P = 0.0345) and OR = 2.297 (1.301–4.057, P = 0.0046), respectively. We also observed that the MCP-1 AG and GG -genotypes contribute to the occurrence of thrombocytopenia in SLE patients OR = 2.618 (1.280–5.352, P = 0.0089). Our observations indicate that either MCP-1 −2518 G variant can be associated with some clinical findings in patients with SLE

Evaluation of osteoprotegerin level and selected inflammatory markers in patients with essential hypertension

Wstęp W ostatnich badaniach wykazano, że stężenie osteoprotegryny (OPG) w surowicy wzrasta w związku z postępem zmian miażdzycowych tętnic wieńcowych, chorobą wieńcową, zawałem serca i przyszłymi chorobami układu sercowo-naczyniowego. Celem pracy było oznaczenie stężenia OPG i poszukiwanie związku między OPG i wybranymi markerami zapalnymi u pacjentów z nadciśnieniem tętniczym. Materiał i metody Do badania włączono 30 pacjentów z samoistnym nadciśnieniem tętniczym. Grupę kontrolną stanowiło 10 zdrowych osób. Przeprowadzono pełne badanie kliniczne. U wszystkich uczestników badania oznaczono stężenie OPG w surowicy metodą immunoenzymatyczną, stężenie w surowicy wysokoczułego białka C-reaktywnego (hsCRP) - metodą turbidymetrii i czynnika martwicy nowotworów &#945; (TNF-&#945;) - metodą radioimunnometryczną. Wyniki U pacjentów z samoistnym nadciśnieniem tętniczym stężenia OPG, hsCRP oraz TNF-&#945; w surowicy krwi znamiennie statystycznie przewyższały wartości obserwowane u osób z grupy kontrolnej (p < 0,05). Stwierdzono również dodatnią korelację między stężeniami OPG i TNF-&#945; (p < 0,05). Wnioski Nadciśnienie tętnicze cechuje nasilony proces zapalny. Wzajemna relacja stężenia OPG i stężenia TNF-&#945; może stanowić wyjaśnienie współwystępowania kalcyfikacji tętnic wieńcowych i progresji miażdżycy ze stanem zapalnym w tej grupie pacjentów. Nadciśnienie Tętnicze 2010, tom 14, nr 5, strony 375-380.Background Recent studies suggest that serum osteoprotegerin (OPG) levels increase in association with coronary artery calcification, coronary artery disease, stroke and future cardiovascular events. The aim of this study was to determine serum OPG levels and to investigate the relationship between OPG and selected inflammatory markers in hypertensive patients. Material and methods Thirty patients with hypertension were studied. As a control 10 healthy volunteers were used. Physical examination was performed. Plasma OPG concentrations (OPG, enzyme-linked immunosorbent assay), high-sensitivity C-reactive protein (hs-CRP, turbidimetry method) and tumor necrosis factor &#945; (TNF-&#945;, radioimmunoassay) were determined. Results The levels of plasma OPG, hsCRP and TNF-&#945; significantly exceeded those observed in the control group (p < 0.05). Positive correlation between OPG and TNF-&#945; (p < 0.05) was found. Conclusions Hypertension is characterized by increased inflammatory process. The association of elevated OPG with TNF-&#945; may provide a mechanistic link between coronary artery calcification, atherosclerosis progression and inflammation in this group of patients. Arterial Hypertension 2010, vol. 14, no 5, pages 375-380

UCHL1/PGP 9.5 Dynamic in neuro-immune-cutaneous milieu : focusing on axonal nerve terminals and epidermal keratinocytes in psoriatic itch

Psoriasis is an immunogenetic skin disease manifesting as plaque lesions on the skin. Patients with psoriasis frequently suffer from itch, an unpleasant sensation causing a desire to scratch. Psoriatic itch is mainly transmitted by unmyelinated C-fibers; however, the exact molecular mechanism of psoriatic itch is still unexplained. Protein gene product 9.5 (PGP 9.5) is a panneurological marker commonly used for analysis of peripheral peptidergic and nonpeptidergic nerves and identification of cutaneous neuro-immune-endocrine cells. However, some studies suggested that nonneuronal cells, like keratinocytes, may also express PGP 9.5. This phenomenon might be linked with impaired axonal transport, keratinocyte injury, or dysfunctions of neuro-immune-cutaneous connections. The aim of this study was to analyze the expression of PGP 9.5 in psoriatic skin. We observed significantly altered density of PGP 9.5-positive axonal nerve terminals in pruritic lesional (p=0.04) and nonlesional psoriatic skin (p>0.001) compared with controls. In contrast, no significant differences were observed between psoriatic skin without itch and controls. Furthermore, PGP 9.5 expression by suprabasal keratinocytes (SBKs) was significantly increased in itchy skin lesions (p=0.007) compared to skin without itch, and a positive correlation was observed between PGP 9.5 expression and itch intensity (r=0.64; p=0.02). Our findings indicate changes in peripheral innervations and psoriatic keratinocytes, which may influence neuro-immune-cutaneous homeostasis and modulate itch transmission

Decreased expression of 17β-hydroxysteroid dehydrogenase type 1 is associated with DNA hypermethylation in colorectal cancer located in the proximal colon

<p>Abstract</p> <p>Background</p> <p>The importance of 17β-estradiol (E2) in the prevention of large bowel tumorigenesis has been shown in many epidemiological studies. Extragonadal E2 may form by the aromatase pathway from androstenedione or the sulfatase pathway from estrone (E1) sulfate followed by E1 reduction to E2 by 17-β-hydroxysteroid dehydrogenase (HSD17B1), so <it>HSD17B1 </it>gene expression may play an important role in the production of E2 in peripheral tissue, including the colon.</p> <p>Methods</p> <p><it>HSD17B1 </it>expression was analyzed in colorectal cancer cell lines (HT29, SW707) and primary colonic adenocarcinoma tissues collected from fifty two patients who underwent radical colon surgical resection. Histopathologically unchanged colonic mucosa located at least 10-20 cm away from the cancerous lesions was obtained from the same patients. Expression level of <it>HSD17B1 </it>using quantitative PCR and western blot were evaluated. DNA methylation level in the 5' flanking region of <it>HSD17B1 </it>CpG rich region was assessed using bisulfite DNA sequencing and HRM analysis. The influence of DNA methylation on HSD17B1 expression was further evaluated by ChIP analysis in HT29 and SW707 cell lines. The conversion of estrone (E1) in to E2 was determined by electrochemiluminescence method.</p> <p>Results</p> <p>We found a significant decrease in HSD17B1 transcript (<it>p </it>= 0.0016) and protein (<it>p </it>= 0.0028) levels in colorectal cancer (CRC) from the proximal but not distal colon and rectum. This reduced <it>HSD17B1 </it>expression was associated with significantly increased DNA methylation (<it>p </it>= 0.003) in the CpG rich region located in the 5' flanking sequence of the <it>HSD17B1 </it>gene in CRC in the proximal but not distal colon and rectum. We also showed that 5-dAzaC induced demethylation of the 5' flanking region of <it>HSD17B1</it>, leading to increased occupation of the promoter by Polymerase II, and increased transcript and protein levels in HT29 and SW707 CRC cells, which contributed to the increase in E2 formation.</p> <p>Conclusions</p> <p>Our results showed that reduced <it>HSD17B1 </it>expression can be associated with DNA methylation in the 5' flanking region of <it>HSD17B1 </it>in CRC from the proximal colon.</p

Impacts of soil conditions and light availability on natural regeneration of Norway spruce Picea abies (L.) H. Karst. in low-elevation mountain forests

& Key message Natural regeneration of P. abies (L.) H. Karst. may reach high densities in lower mountain elevations. The highest densities were found in sites with moderate light availability, with low pH, and not near the riverbank. However, age-height classes differed in the predicted magnitude of response, but were consistent in response directions. Mosses and understory species typical of coniferous forests were positively correlated with regeneration density. & Context Norway spruce Picea abies (L.) H. Karst. in Central Europe is at risk under climate change scenarios, particularly in mountain regions. Little is known about the impact of environmental factors on the natural regeneration of P. abies in lowelevation mountain forests. & Aims We aimed to assess impacts of distance from the riverbank, soil pH, and light availability on natural P. abies regeneration. We hypothesized that (1) natural P. abiesregeneration would depend on light availability and soil pH and (2) there are understory plant species which may indicate the microsites suitable for natural regeneration of P. abies. & Methods The study was conducted in the Stołowe Mountains National Park (SW Poland, 600–800 m a.s.l.). We established 160 study plots (25 m2 ) for natural regeneration, light availability, soil pH, and understory vegetation assessment