A novel report of apoptosis in human lung carcinoma cells using selective agonist of D2-like dopamine receptors: a new approach for the treatment of human non-small cell lung cancer.

In our previous study, a relationship between low expression of D2-like dopamine receptor genes and non-small cell lung cancer (NSCLC) disease was found. In this new research, by using selective agonist of these receptors, Bromocriptine (BR), we attempted to activate D2-like expression and apoptotic induction in a selective cell line of NSCLC. In addition, the relationship of apoptotic response of human lung carcinoma cells to BR and D2- dopamine receptor genes is investigated. Human lung cancer (QU-DB) cells were treated by five doses of BR at 48 h and cell viability was determined by MTT assay. The gene expression pattern of D2-like dopamine receptor Genes was studied by Real Time PCR. Nuclear morphology of cells was monitored by DAPI flourescent staining then induction of DNA fragmentation by BR was shown in an agarose gel. Finally, the detection and quantification of apoptosis and its differentiation from necrosis was carried out by using Annecxin-V-Fluos Staining. In this study, it is demonstrated that BR inhibited the proliferation of human lung cancer cells and induced apoptosis in them. In addition, the probable relationship between D2-dopamine receptor genes expression and the development of apoptosis was found. In conclusion, BR is responsible for induction of apoptosis in human lung cancer cells and can be used in treatment of these tumoric cells. In addition, normal expression of D2 dopamine receptors was associated with apoptotic effect of BR on these cells.

Plasma glucose lowering effect of the wild Satureja khuzestanica Jamzad essential oil in diabetic rats: role of decreased gluconeogenesis.

This study was to evaluate the effect of the wild SKEO on activities and genes expression of hepatic Glycogen Phosphorylase (GP) and phosphoenolpyruvate carboxykinase (PEPCK) in normal and diabetic rats. The wild SKEO was orally administered at different doses (50 and 100 mg/kg/day) to normal as well as diabetic rats for 21 days. The levels of mRNA were determined using the quantitative real-time RT-PCR technique. The plasma glucose concentrations of diabetic rats receiving SKEO (100 mg kg(-1)) compared with diabetic control were significantly decreased. Hepatic GP activity and its mRNA levels of diabetic rats treated with SKEO moderately increased. The activity of hepatic PEPCK and its mRNA levels were significantly decreased in normal rats treated with SKEO (100 mg kg(-1)). The enhancement of PEPCK activity and its mRNA levels of diabetic treated rats with SEKO (100 mg kg(-1)) was significantly decreased compared with diabetic control. In conclusion, an excessive inhibition of PEPCK in liver of diabetic rats treated with the wild SKEO may contribute to the plasma glucose lowering action of SKEO that seems to be in relation with antioxidant properties of SKEO.

A novel tissue engineering-based assay for immunological infertility.

Antisperm antibodies (ASA) are present in 9-36% of infertile couples, a condition called immunological infertility. The variability of ASA in terms of antigenic specificity and biological effects has made it difficult to design a test able to distinguish reliably between ASA that contribute to infertility and those that do not. To develop a reliable and reproducible method able to detect sperm antibodies, we took advantage of recent progress made in tissue engineering techniques. We used collagen gel as a bio-scaffold for the production of engineered sperm analogues. The advantages of using collagen gels include biocompatibility, ease of fabrication and low cost. We found that this tissue engineering-based assay is more specific and more sensitive than a conventional test routinely used for ASA detection. In addition, it exhibited low intra- and inter-variations. We envision the use of this novel approach for the detection of a variety of autoantibodies in autoimmune diseases. In addition to diagnostic purposes, tissue-engineering based tests could be useful in monitoring treatments with bio-drugs.

Growth hormone antibodies formation in patients treated with recombinant human growth hormone.

Human growth hormone (hGH) is normally produced by acidophilic cells of the anterior lobe of the pituitary gland. Recombinant DNA technology has made it possible to produce rhGH. There have been reports of immunological reactions in patients treated with rhGH. For this reason, it is necessary to check sera of patients for presence of antibody against rhGH. Forty-seven children were treated for up to 6 months with recombinant human growth hormone (rhGH-Novo), 0.1 IU/Kg body weight, subcutaneously, three times weekly. The magnitude of growth response was similar to those expected from clinical experience with pituitary growth hormone. We examined sera for specific antibodies against rhGH by ELISA methods. Four patients developed serum antibodies against growth hormone. The analysis of these four sera by Dot blotting method also showed presence of antibodies against rhGH. In the sera of treated patients, pre-incubated with different concentration of rhGH, specific antibodies were detected by neutralizing assay. This finding was confirmed by ELISA technique. In conclusion, the main concern with anti-GH antibodies could be their ability to neutralize circulating growth hormone and inhibition its growth promoting effect.

The expression of the gene coding for the antibacterial peptide LL-37 is induced in human keratinocytes during inflammatory disorders.

The epithelia constitute a major barrier to the environment and provide the first line of defense against invading microbes. Antimicrobial peptides are emerging as participants in the defense system of epithelial barriers in general. Originally we isolated the human antimicrobial peptide LL-37 from granulocytes. The gene (CAMP or cathelicidin antimicrobial peptide) coding for this peptide belongs to the cathelicidin family, whose members contain a conserved pro-part of the cathelin type. The human genome seems to have only one gene of this family, whereas some mammalian species have several cathelicidin genes. In the present work we demonstrate up-regulation of this human cathelicidin gene in inflammatory skin disorders, whereas in normal skin no induction was found. By in situ hybridization and immunohistochemistry the transcript and the peptide were located in keratinocytes throughout the epidermis of the inflammatory regions. In addition, the peptide was detected in partially pure fractions derived from psoriatic scales by immunoblotting. These fractions also exhibited antibacterial activity. We propose a protective role for LL-37, when the integrity of the skin barrier is damaged, participating in the first line of defense, and preventing local infection and systemic invasion of microbes.

RT-PCR topography of chronic psoriasis skin based on analysis of T-cell receptor B variable region gene usage.

Psoriasis is a hyperproliferative inflammatory disease and 70% of patients develop a chronic plaque form. The pathogenesis of psoriasis is not known but evidence exists that T cells play a crucial role. The T cell V-gene receptor repertoire from psoriasis skin (different layers) was compared with peripheral blood T cells by employing RNA polymerase chain reaction (PCR) amplification. T cell receptor (TCR) BV 5.1, 11, 12, 13.1 and 16 were utilized to a significantly higher degree in areas close to the basal layers when compared to CD4+, CD8+ or unfractionated blood T cells from the same patients, whereas only BV11 and 13.1 genes of T cells from deeper layers of the dermis showed such a skewed usage. No biased usage of TCRBV genes was observed in superficial layers or in whole skin. Furthermore, T cell receptor junctional diversity analysed by high resolution gel electrophoresis showed skin psoriatic T cells to be poly- or oligoclonal. In conclusion, we show that TCRBV gene usage from different layers of psoriatic skin has a different pattern compared with the corresponding gene usage in circulating peripheral blood T cells. This pattern may implicate possible skin-associated antigen or superantigens activating a limited number of T cells in areas of skin close to basal layers, which in turn could promote keratinocyte proliferation.

RT-PCR based analysis of T-cell receptor B variable region gene usage in normal human breast skin resident T lymphocytes (SRT).

The skin interfaces directly with the external environment that contains innumerable infectious agents. Therefore, an appropriate and rapid immunologic response is required to preserve internal homeostasis. An essential feature of the "skin immuno system' (SIS) is the presence of substantial numbers of T cells in normal skin. The T-cell receptor repertoire from normal human breast skin was analysed quantitatively and qualitatively by using PCR amplification of reverse transcribed RNA, T-cell receptor BV3 and BV14 gene usage was increased in skir T lymphocytes in all individuals tested (n = 8) compared to peripheral blood CD4+ and CD8+ T lymphocytes from the same individuals. The T-cell receptor junctional diversity analysed by high resolution gel electrophoresis showed skin T-cell BV3 and BV14 gene usage to be predominantly polyclonal. Superantigen stimulation of T cells in human skin is considered a likely explanation of the present finding.