CD55, CD59, factor H and factor H-like 1 gene expression analysis in tumors of the ovary and corpus uteri origin.

The expression level of complement regulators in ovarian and corpus uteri tumors was not fully established so far. In current manuscript we performed gene expression analysis by the real-time PCR approach to investigate both membrane bound - CD55 and CD59 and fluid phase - factor H and factor H-like 1 complement regulators. We found increased CD55 expression in corpus uteri tumors when compared to control tissues, whereas in ovarian cancer CD55 expression was lower than in control sections. Additionally we found CD59 expression to be more prominent in ovarian cancer than in corpus uteri tumor samples. We observed also the strong positive correlation between the level of expression of the whole group of regulators, which was particularly significant between the expression of factor H and factor H- like 1. In conclusion we present novel results which implicates different role of particular complement inhibitors in the regulation of the complement system in two cancer types examined. Strong positive correlation between examined proteins implicates similar pattern of the regulation which should be taken into consideration with regards to the possible immunotherapy applied as adjuvant therapeutic approach in these two indications. The inhibition of complement regulation may serve as a strategy to potentiate the efficacy of such treatment.

NADPH oxidase and uncoupled nitric oxide synthase are major sources of reactive oxygen species in oral squamous cell carcinoma. Potential implications for immune regulation in high oxidative stress conditions.

The development of cancer is associated with high oxidative stress and at the same time with immune system activation. Tumors develop efficient mechanisms of protection against the immune response, which allow them to escape the immune surveillance. Simultaneously, key events in the process of carcinogenesis are related to oxidative stress. The relationship between the two remains unknown. Novel understanding of oxidative stress shows that discrete changes of activities of certain enzyme systems such as NADPH oxidases or nitric oxide synthases may be more important than the overall balance of production and removal of reactive oxygen species. Such imbalance of nitric oxide and superoxide production could modify inflammation and immune regulation. We studied superoxide anion production (by lucigenin enhanced chemiluminescence - 5 microM), NADPH oxidase activity and nitric oxide synthase (NOS) dysfunction. In parallel mRNA expression of immunomodulatory markers such as FoxP3 (T regulatory cell marker), CCR6 (mucosal homing effector T cell marker) and CD85j (NK cell/CD8 T cell Ig-like MHC class I inhibitory receptor) was determined. Basal superoxide production and NADPH oxidase activity are increased in oral squamous cell carcinoma. Tumor superoxide production was inhibited by NADPH oxidase inhibitor apocynin and by NOS inhibitor L-NAME. This indicates, for the first time, that oral squamous cell carcinoma is characterized by dysregulated nitric oxide synthase, which apart from increased NADPH oxidase activity contributes to oxidative stress and may be related to the immuno-pathology of these tumors. Studied tumors were infiltrated by CCR6+, but showed lower expression of both CD85j and FoxP3 mRNA. Finally, the CD85j mRNA expression was inversely correlated to oxidative stress parameters. These preliminary studies indicate that tumor oxidative stress, related to NADPH oxidase activity and NOS activity could be related to immune responses to cancer, thus therapeutic modification of oxidative stress, which could include the correction of NOS dysfunction, could facilitate immune surveillance.

Efficiency of lipofection of adherent cells is limited by apoptosis.

Stability of gene expression and transfection efficiency plays the main role in the application of gene transfer method. In somatic cell gene delivery, expression of the gene product is limited by the function of the cell to which it is delivered. In the present study analyzing the lipofected adherent cells, we have shown that lower level of transgene: beta-galactosidase activity at later time period correlated with decrease in cell viability, which was shown to be due to apoptosis. Apoptosis following DNA uptake occurred only when DNA was present during lipofection.

[A case of Wilson's disease with an atypical clinical course]. / Przypadek choroby Wilsona o nietypowym przebiegu.

The case of atypical course of Wilson's disease in a 31 years old woman was reported. Basing on its clinical presentation it was classified as a liver form. The laboratory criteria necessary for the diagnosis of this disease are as follows: increase of cooper concentration in liver tissue (above 500 micrograms copper per gram dry substance), defect of copper incorporation into ceruloplasmin in 24 hours copper excretion test in urine (above mumol/24 h). In this report the main principles of pharmacotherapy were summarized, with particular stress on the necessity throughout the patient's life.

A rapid cold-hardening response protecting against cold shock injury in Drosophila melanogaster.

In studies of insect cold-hardiness, the supercooling point (SCP) is defined as the temperature at which spontaneous nucleation of body fluids occurs. Despite having an SCP of -20 degrees C, adults of Drosophila melanogaster did not survive exposure to -5 degrees C, which suggests that cold shock causes lethal injury that is not associated with freezing. If, however, flies were chilled at 5 degrees C, for as little as 30 min, approximately 50% of the flies survived exposure to -5 degrees C for 2h. This capacity to cold-harden rapidly was greatest in 3- and 5-day-old adults. The rapid cold-hardening response was also observed in larvae and pupae: no larvae survived 2 h of exposure to -5 degrees C, whereas 63% pupariated if chilled at 5 degrees C before subzero exposure. Similarly, although exposure of pupae to -8 degrees C was lethal, if pre-chilled at 5 degrees C 22% eclosed. This extremely rapid cold-hardening response may function to allow insects to enhance cold-tolerance in response to diurnal or unexpected seasonal decreases in environmental temperature.

Binedaline binding to plasma proteins and red blood cells in humans.

Serum binding of binedaline, a new antidepressant drug, was studied in vitro by equilibrium dialysis. The percent of binding in serum is high, 99.2%, and remains constant within the range of therapeutic concentrations; no saturation to the binding sites was seen. Investigations performed on isolated proteins with a wide range of concentrations showed one site with a high affinity constant (Ka = 2 X 10(6) M-1) for alpha 1-acid glycoprotein and two sites with a low affinity constant (Ka = 3 X 10(4) M-1) for human serum albumin. Binding to lipoproteins was nonsaturable, with a total affinity constant of 1.25 X 10(5) less than nKa less than 2.79 X 10(6) M-1. Over the range of therapeutic concentrations, the ratio of binedaline concentrations in serum and red blood cells remained constant (1%) and was shown to be dependent on the free fraction of binedaline in serum.

Sister chromatid exchange frequency in mouse embryo chromosomes after in utero ethanol exposure.

The frequency of sister chromatid exchange (SCE) increased in mouse embryo chromosomes following a single maternal i.p. injection of ethanol on the tenth day of gestation, indicating a possible morphogenetic action of ethanol in the developing mouse embryo.

An organ culture model for study of biochemical development of fetal rat lung.

We have developed a short-term organ culture model for the study of the biochemical and morphological development of late gestation fetal rat lung. Explants (1 mm3) of 19-day lung were cultured in an oxygen enriched environment in the presence of synthetic serum-free medium for 3 days. Morphological maturation continued in culture. The rate of incorporation of choline into disaturated phosphatidylcholine and the content of this phospholipid in the explants increased in vitro in a pattern very similar to that which occurs in vivo. The activities of choline kinase and cholinephosphotransferase were also similar in cultured lung and in vivo. Studies of glucose oxidation to CO2 provided additional evidence that the explants remained viable in culture. The explants retained the sensitivity of fetal lung to hormonal action. This was demonstrated by the stimulation of choline incorporation into phospholipid by cyclic AMP and an increase in the glycogen content after exposure to insulin.