Osteogenic differentiation of human adipose-derived stem cells in 3D conditions - comparison of spheroids and polystyrene scaffolds.

Expansion and differentiation of adipose-derived stem cells (ADSCs) in vitro are routinely performed in two-dimensional (2D) environments. The study hypothesis was that the utilisation of three-dimensional (3D) culture conditions, mimicking the natural stem cell niche, might increase the osteogenic commitment of ADSCs. Therefore, human ADSCs were seeded in 3D culture systems lacking bioactive material components: spheroids and polystyrene scaffolds. ALP activity, a marker of early osteogenesis, was higher in ADSC spheroids and ADSC seeded on polystyrene scaffolds as compared to 2D cultures. Furthermore, the expression of the osteoblast marker genes Runt-related transcription factor 2 (RUNX2), osterix and integrin binding sialoprotein (IBSP) was significantly up-regulated in spheroids as compared to polystyrene scaffolds and 2D culture. Elevated levels of RUNX2 and IBSP in spheroids were confirmed at the protein level by Western blot and immunofluorescence, respectively. Bone mineral production was lower in spheroids than in polystyrene scaffolds and 2D culture at day 14. Curiously, adipocyte differentiation was downregulated in spheroids as compared to 2D-culture. Finally, to induce late differentiation events, cells were dissociated from spheroids after a 7 d osteogenic pre-differentiation culture and replated in 2D culture in osteoblast maturation medium. After a subsequent 14 d of maturation, cells produced bone mineral and osteocalcin proteins, which are late osteoblast markers. The present work showed that the 3D environment may provide additional stimuli for the commitment of ADSCs to the osteogenic lineage. Furthermore, the presented data may be valuable when designing protocols to prepare ADSCs for use in bone regeneration clinical studies.

Uroplakin IIIa Is a Marker in Bladder Cancer but Seems Not to Reflect Chemical Carcinogenesis.

BACKGROUND: Uroplakins are glycoproteins investigated as potential markers of urothelial carcinoma. However, their role in chemical carcinogenesis is uncertain. In this study the diagnostic value of plasma and urine uroplakin IIIa (UPIIIa) levels in bladder cancer (BC) was investigated, particularly in the aspect of environmental exposure to chemical carcinogens, measured by DNA damage and detoxification ability in the BC smoking group. The correlation between uroplakin, 8-OHdG, and GSTπ was investigated. MATERIAL AND METHODS: This study included 61 BC patients and 33 healthy controls. UPIIIa, 8-OHdG, and GSTπ levels were estimated by the immunoenzymatic method (ELISA). RESULTS: UPIIIa levels were elevated in BC patients in plasma (p≤0.001) and in urine (p≤0.001), as were 8-OHdG and GSTπ levels in urine. Moreover, the 8-OHdG level was higher in invasive or high grade tumors. A positive correlation between UPIIIa/GSTπ and 8-OHdG/GSTπ was observed, but no UPIIIa/8-OHdG correlation was noted. CONCLUSION: The study showed the diagnostic value of urine and plasma UPIIIa in BC (good sensitivity, specificity, and predictive value). The lack of UPIIIa correlation with 8-OHdG and smoking suggests that UPIIIa does not reflect the environmental exposure. The increased levels of 8-OHdG and GSTπ in the invasive tumor stage indicate their value in BC monitoring.

Preliminary Evaluation of the Diagnostic Usefulness of Uroplakin 2 with an Assessment of the Antioxidant Potential of Patients with Bladder Cancer.

BACKGROUND: Urothelial carcinoma is the most common type of bladder cancer (BC). It makes up more than 90% of all bladder cancers. Uroplakins are tissue-specific, glycoproteins, playing a role in the construction and function of urothelium. The emergence of uroplakins in the urine and/or plasma may be of potential importance in the early detection of BC. In our study, the diagnostic value of plasma and urine uroplakin 2 (UP2) concentration in bladder cancer was investigated, with an assessment of the antioxidant potential of BC patients. The correlation between UP2, total antioxidant capacity (TAC), and concentration of glutathione (GSH) was also examined. MATERIALS AND METHODS: This study included 61 BC patients and 33 healthy controls. UP2 concentration was estimated by the immunoenzymatic method (ELISA). TAC and GSH were determined in spectrophotometrically methods. RESULTS: UP2 concentration in BC patients was significantly higher (p≤0.001) both in plasma and in urine compared to the control groups (C). TAC concentration in urine (p≤0.001) and GSH concentration in plasma (p=0.047) were significantly lower in BC group compared to the C group. The high specificity and sensitivity for UPK2 in plasma (76%, 80%, respectively) and urine (88%, 84%, respectively) were observed. Positive correlations were observed between concentration of UP2 in plasma and TAC concentration in urine and between UP2 concentration in plasma and GSH concentration in the same material. CONCLUSION: The study showed the early diagnostic value of urine and plasma UP2 in BC. There was a decrease in UP2 concentration in the urine of patients with the development of BC. The decrease of antioxidant systems (TAC, GSH) indicates their relationship with the BC process. Based on the obtained results, it is justified to continue the study in a larger group of patients with BC.

Increased serum levels of lipopolysaccharide and antiflagellin antibodies in patients with diarrhea-predominant irritable bowel syndrome.

BACKGROUND: Innate immune responses to conserved microbial products such as lipopolysaccharide (LPS) and flagellin are likely important in microbial-host interactions and intestinal homeostasis. We hypothesized that bacterial translocation and activation of mucosal immunity against common microbial antigens might be involved in the development of irritable bowel syndrome (IBS). We therefore compared serum levels of LPS, soluble CD14 (sCD14), and flagellin antibodies between patients with different subtypes of IBS and healthy controls. METHODS: We analyzed serum obtained from 88 patients (74 females) aged 19(43)-73 years and 106 healthy volunteers (77 females) aged 19(38)-62 years. Diarrhea-predominant IBS (D-IBS) was present in 32 patients (36%), 23 patients (26%) had constipation-predominant IBS (C-IBS), and 33 patients (38%) had A-IBS. We used ELISA for sCD14 and antiflagellin immunoglobulin G and limulus amebocyte assay for LPS. Abdominal symptoms and psychiatric comorbidities were assessed using validated questionnaires. KEY RESULTS: We found a significantly higher serum level of LPS in patients with D-IBS compared to controls (p = 0.0155). The level of antibodies to flagellin was higher in patients with IBS than in controls (mainly driven by higher levels in D-IBS, p = 0.0018). The levels of sCD14 were lower in D-IBS patients compared to controls (p = 0.0498). We found a weak, but significant correlation between the levels of antiflagellin antibodies and anxiety among IBS patients (ρ = 0.38; p = 0.0045). CONCLUSIONS & INFERENCES: Our results support the concept that immune reactivity to luminal antigens may have a role in the development of D-IBS. The serum level of antiflagellin antibodies was found to correlate with patients' self-reported anxiety score.

Hedgehog/GLI and PI3K signaling in the initiation and maintenance of chronic lymphocytic leukemia.

The initiation and maintenance of a malignant phenotype requires complex and synergistic interactions of multiple oncogenic signals. The Hedgehog (HH)/GLI pathway has been implicated in a variety of cancer entities and targeted pathway inhibition is of therapeutic relevance. Signal cross-talk with other cancer pathways including PI3K/AKT modulates HH/GLI signal strength and its oncogenicity. In this study, we addressed the role of HH/GLI and its putative interaction with the PI3K/AKT cascade in the initiation and maintenance of chronic lymphocytic leukemia (CLL). Using transgenic mouse models, we show that B-cell-specific constitutive activation of HH/GLI signaling either at the level of the HH effector and drug target Smoothened or at the level of the GLI transcription factors does not suffice to initiate a CLL-like phenotype characterized by the accumulation of CD5(+) B cells in the lymphatic system and peripheral blood. Furthermore, Hh/Gli activation in Pten-deficient B cells with activated Pi3K/Akt signaling failed to enhance the expansion of leukemic CD5(+) B cells, suggesting that genetic or epigenetic alterations leading to aberrant HH/GLI signaling in B cells do not suffice to elicit a CLL-like phenotype in mice. By contrast, we identify a critical role of GLI and PI3K signaling for the survival of human primary CLL cells. We show that combined targeting of GLI and PI3K/AKT/mTOR signaling can have a synergistic therapeutic effect in cells from a subgroup of CLL patients, thereby providing a basis for the evaluation of future combination therapies targeting HH/GLI and PI3K signaling in this common hematopoietic malignancy.

Infection of human enteroendocrine cells with Chlamydia trachomatis: a possible model for pathogenesis in irritable bowel syndrome.

BACKGROUND: Irritable bowel syndrome (IBS) is a widespread gastrointestinal disorder of unknown etiology. Recently, our group detected chlamydial antigens in enteroendocrine cells (EEC) of jejunum biopsies from patients with IBS. Impairment of EEC secretion upon Chlamydia infection might lead to disturbances of gut functions. We have therefore studied the interaction between Chlamydia and EEC in vitro. METHODS: Two different human enteroendocrine cell lines were studied: LCC-18 from a neuroendocrine colonic tumour and CNDT2 from a small intestinal carcinoid. Cell lines were infected with C. trachomatis serovar LGV II strain 434. We used Penicillin G for inducing persistent infection. The ultrastructure of infected cells was studied using transmission electron microscopy and immunofluorescence and we used RT-PCR analysis for studying changes in gene expression at different stages of infection. KEY RESULTS: We found that both cell lines could be infected with C. trachomatis yielding productive infections and persistence could be induced using penicillin G. Immunofluorescence showed different cellular distributions of serotonin and chromogranin A in non-infected (cytoplasmatic distribution) compared with infected cells (serotonin and chromogranin mostly in chlamydial inclusions). In line with the microscopical findings, we found a significant down-regulation of the gene coding for the vesicular monoamine transporter (VMAT1) in infected compared with non-infected EEC (P<0.05). CONCLUSIONS & INFERENCES: Altered protein distributions together with down-regulation of VMAT1 suggest that chlamydial infection may influence vesicular transport. It is therefore possible that such an infection in vivo could lead to disturbances in the regulation of gut functions.

Enzymuria and low molecular weight protein excretion as the differentiating marker of complications in the early post kidney transplantation period.

Renal function in the early post-transplantation period depends largely on factors affecting the kidney prior to implantation. Function of the graft may be also disturbed by the most common complications of the early post-operative period such as acute graft rejection (AGR), acute tubular necrosis (ATN) and may be modified by nephrotoxic action of cyclosporine A (CsA). Evaluation of excretion of enzymes and low molecular weight proteins (LMWP) may help in the differentiation of these complications. Aim Comparison of the urinary excretion of markers of tubular injury in patients with AGR, ATN, or patients with stable graft function (SGF) was made and differences between groups and correlations between markers and cold ischemia time (CIT), warm ischemia time (WIT) and blood trough level of cyclosporine A (CsA0) were determined. Material and methods In 60 cadaveric renal allograft recipients in the early post-transplantation period urinary excretion of N-acetyl-beta-D-glucosaminidase (NAG) and B isoenzyme (NAG-B), alanylaminopeptidase (AAP), gamma-glutamyltransferase (GGT), alpha and pi isoenzymes of glutathione S-transferase (alpha-GST, pi-GST), retinol binding protein (RBP) and beta2- microglobulin (beta2M), were analyzed. Results NAG and NAB-B activities were higher in ATN (P<0.05, P<0.01) and in AGR (P<0.005, P<0.02) than in SGF. Excretion of pi-GST was higher in AGR than in SGF (P<0.0002) or ATN (P<0.007). CIT and WIT in patients with ATN were higher (P<0.05) than in SGF group. In ATN patients, correlations of CIT with RBP (P<0.05) and pi-GST (P<0.05), and WIT with RBP (P<0.05), and pi-GST (P<0.001) were found. Conclusions High urinary NAG and NAG B excretion characterizes ATN and AGR patients. Evaluating urinary excretion of pi-GST may be helpful in differentiating AGR from ATN. However, taking into account ischemia time is necessary in interpreting the pi-GST value in early post transplant period.

Styrene and ethylene glycol have a synergetic effect on lipid peroxidation that is better protected than repaired by CoQ10.

Previous study of a group of 22 workers occupationally exposed to styrene, ethylene glycol and their mixture at a paint and lacquer industry indicated significantly elevated concentration of malondialdehyde with 4-hydroxynonenal (MDA+4-HNE) in the blood plasma, successfully decreased with coenzyme Q10 (CoQ10) supplementation. The aim of present study is to evaluate whether the exposure to styrene or/and ethylene glycol could be responsible for the increase in MDA level. The mechanism of a single solvent action and the mixture was examined, specially whether it is connected with hydroxyl radical (*OH) generation. It was also investigated whether coenzyme Q10 could be considered as a protective (given before the solvents) or repairing (given after the solvents) agent in oxidative stress caused by the solvents. The results indicate that ethylene glycol nor styrene increase MDA and *OH, but as a mixture give synergetic interaction, elevating MDA and *OH concentration to a statistically significant extent. Coenzyme Q10 at a dose of 3.0 microg/ml only protects, but does not repair increased lipid peroxidation caused by ethylene glycol with styrene. In order to obtain both a protective and repairing effect, a concentration of 12.0 microg/ml CoQ is needed.

Oxidative stress and coenzyme Q10 supplementation in renal transplant recipients.

Recently some reports about the oxidative stress in renal transplant recipients have been published. The role of coenzyme Q10 (CoQ10) as radical scavanger is largely known. The aim of our study was to evaluate the protective role of CoQ10 in renal transplant recipients on lipid peroxidation and lipids parameters, as well as its influence on antioxidant enzymes, neutrophils chemiluminescence and urinary enzymes. The study was performed in 11 long term allograft recipients treated additionally with CoQ10 90 mg/day in three doses, 30 mg each for four weeks. The malonyldialdehyde (MDA) and 4-hydroxynonenal (4-HNE), superoxide dismutase (SOD) and glutathione peroxidase (GPx) and the basic parameters of lipid metabolism such as total cholesterol (TC), high and low density lipoproteins (HDL, LDL), triglycerides (TG), atherogenicity indicators [LDL/HDL; (TC-HDL)/HDL] were evaluated. The chemiluminescence of neutrophils (luminol, fLMP-method) were mesured and the activity of N-acetyl-beta-D-glucosaminidase (NAG), alanylaminopeptidase (AAP), elastase, alpha-1-antitrypsin. All parameters were estimated before and after CoQ10 treatment. Statistically significant changes were noticed with the LDL and atherogenicity indicators (p < 0.01) (decrease) as well as HDL level (p < 0.001) (increase). Also the significant decrease of fMLP stimulated PMNL chemiluminescence (p < 0.05) confirms the antioxidative properties of CoQ10. The significant increase of NAG activity (p < 0.05) can't be the result of nephrotoxic effect, because NAG-B is unchanged. Serum concentration of creatinine and cyclosporine A in renal allograft recipients was unchanged after CoQ10 treatment. The presented date shows that further study with CoQ10 treatment in renal transplant in larger numbers and over longer periods should be considered.

Lipid peroxidation stimulated by Solvesso, Bawanol and methanol, and its counteraction by antioxidants in human placental mitochondria.

The influence of some solvents used in the paint and lacquer industry, Solvesso 100, 150 and Bawanol, on the process of lipid peroxidation was evaluated. The interactions of the solvents with methanol and the antioxidative effect of coenzyme Q(10) (CoQ(10)) and vitamin E were also examined. The mitochondria isolated from human placenta were used as a model for investigation. The level of peroxidation was evaluated by malonodialdehyde (MDA) measurement. It was observed that Solvesso 150, Bawanol and methanol caused a statistically significant increase of MDA concentration and that the examined solvents had an antagonistic interaction with methanol. Solvesso 100 does not influence on MDA concentration but gives the same type of interaction with methanol as Solvesso 150. CoQ(10) and vitamin E proved to be effective antioxidants to counteract oxidative stress caused by the studied solvents, lowering lipid peroxidation expressed by MDA concentration.

Antioxidative properties of coenzyme Q10 and vitamin E in exposure to xylene and gasoline and their mixture with methanol.

Exposure to a mixture of solvents in industry is still a problem particularly in industrial laboratories. In the paint and laquer industry the employees are exposed to xylene (Rx) and gasoline (Rg). The influence of xylene and gasoline or their mixture with methanol on lipid peroxidation was evaluated in the presented paper. Antioxidative properties of CoQ10 or vitamin E were also tested. It was observed that xylene caused an increase of lipid peroxidation measured as a MDA level in all used concentrations, but gasoline only in very high doses. The mixture of xylene with methanol increased significantly MDA level, whereas gasoline with methanol did not influence lipid peroxidation. The character of interaction depends on hydrocarbons dose. CoQ10 and vitamin E are effective antioxidants lowering the increased MDA level caused by xylene, gasoline or their mixture with methanol, however the dose of CoQ10 should be adjusted to the strength of oxidative stress in order to avoid disadvantageous effect. CoQ10 is a more effective antioxidant in exposure to xylene rather than gasoline, but vitamin E acts better in exposure to gasoline decreasing the MDA level.

[Environmental exposure of the Wroclaw inhabitants to nickel and assessment of its nephrotoxic effects]. / Srodowiskowe narazenie na nikiel mieszkanców Wroclawia i ocena jego dzialania nefrotoksycznego.

The aim of the research was to evaluate the environmental exposure of Wroclaw inhabitants on nickel compounds and an assessment of their nephrotoxic effects. The study was realized in a group of 74 inhabitants of the Wroclaw's district Krzyki and compared with a group of 24 inhabitants of the countryfield Kobierzyce community located in the 20 km distance from Wroclaw. A nickel concentration was measured in morning urine of all the Wroclaw inhabitants, whereas nephrotoxicity indices were determined in the 26 inhabitants characterised by the increased nickel concentration (2.8 micrograms/l); in urine of 19 inhabitants this concentration was found to be normal. The average nickel concentration in the urine of the 74 Wroclaw inhabitants was equal to 2.85 +/- 0.22 micrograms/l. 40.5% of this group were characterised by the concentration higher than the physiological one, 4.05% of this group had that concentration higher than 7.9 micrograms/l which gives an evidence of the environmental exposure. In the Kobierzyce group the average nickel concentration in urine was 3.34 +/- 0.23 micrograms/l which is significantly higher (at p < or = 0.001) than that for the Wroclaw group. Moreover, in the Kobierzyce group, 62.5% inhabitants was higher the nickel concentration than the physiological one but still lower than the environmental exposure. Our results indicate that the Wroclaw inhabitants are not more exposed to nickel compounds in comparison with the countryside inhabitants. A nephrotoxic action of nickel was detected through the increased excretion of the sialic acid in urine which is an evidence of the reduced glomelural filtration. A positive correlation between the urine nickel concentration and the urine free sialic acid was found (r = 0.61). No increase in the urine N-acetyl-beta-D-glucosaminidase (NAG), isoenzyme NAG-B, beta 2-microglobulin (beta 2 M) and retinal binding protein (RBP) concentrations were found which could indicate a damage in kidney tubules.

Enzymuria and beta2-mikroglobulinuria in the assessment of the influence of proteinuria on the progression of glomerulopathies.

The important role of the tubulo-interstitial system for the progression of glomerulonephritis (GN), is the cause of a continuous search for the proper markers of kidney tubules damage, which can be applied in clinical diagnosis. In the present work the activity of N-acetyl-beta-D-glucosamidase (NAG), its isoenzyme NAG-B, alanylaminopeptidase (AAP), gamma-glutamyltransferase (GGT), concentration of beta2-microglobulin (beta2M) and daily protein excretion in the urine of 37 patients with morphologically different glomerulopathies were measured. The serum creatinine was also controlled. The obtained results suggest that activity of NAG in the patients with GN has an intermediate connection with proteinuria and could be a cause of the inflammatory process of the kidney, but the activity of AAP is directly dependent on urine protein concentration. Systemic analysis of both partial and multiple correlation coefficients of the examined indicators creates new, additional possibilities in the estimation of activity and progress of GN.

Cytotoxicity of some potential DNA intercalators (carbazole, acridine and anthracene derivatives) evaluated through neutrophil chemiluminescence.

A modifying effect of potential DNA intercalators, belonging to a group of carbazole, acridine and anthracene derivatives, on the course of luminol-dependent chemiluminescence of neutrophils (polymorphonuclear leucocytes; PMNL) in the process of phagocytosis was studied. This effect was also examined in reactive-oxygen-species-generating non-cellular reaction systems consisted of myeloperoxidase or xanthine oxidase. Adriamycin (Doxorubicin), which is widely applied to neoplasm therapy, was used as a reference intercalator in the conducted experiments. It was demonstrated that some structurally different derivatives of carbazole inhibited the light emission from N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced neutrophils to the same degree as adriamycin. It can be suggested that the same inhibitory effect was caused by either a different cellular distribution of the derivatives or different interactions of the derivatives with reactive oxygen species in the investigated systems. Measurements of chemiluminescence suggested that the thiol group in one of the carbazole derivatives could strongly interfere with oxidative cell metabolism. In contrast to the analogous derivative of carbazole, both anthracene and acridine derivatives, possessing an N-1'-hydroxyethyl-ethylenodiamino group, induced different increases in chemiluminescence accompanying the process of neutrophil phagocytosis. Cytotoxicity of the investigated derivatives, being tested previously in cancer cells with a sulphorhodamine B assay, was found to possess a specific representation in the complex picture of the derivative-caused modification both of neutrophil and enzymatic non-cellular chemiluminescence. We suggest that chemiluminescence assays may serve as a helpful tool in the primary screening of drug cytotoxicity.

Cross-talk between epidermal growth factor receptor and protein kinase C during calcium-induced differentiation of keratinocytes.

The induction of epidermal differentiation by extracellular Ca2+ involves activation of both tyrosine kinase and protein kinase C (PKC) signaling cascades. To determine if the differentiation-dependent activation of tyrosine kinase signaling can influence the PKC pathway, we examined the tyrosine phosphorylation status of PKC isoforms in primary mouse keratinocytes stimulated to terminally differentiate with Ca2+. Elevation of extracellular Ca2+ induced tyrosine phosphorylation of PKC-delta, but not the other keratinocyte PKC isoforms (alpha, epsilon, eta, zeta). We have previously demonstrated that activation of the epidermal growth factor receptor (EGFR) pathway induces PKC-delta tyrosine phosphorylation in basal keratinocytes (Denning M F, Dlugosz A A, Threadgill D W, Magnuson T, Yuspa S H (1996) J Biol Chem 271: 5325-5331). When basal keratinocytes were stimulated to differentiate by Ca2+, the level of cell-associated transforming growth factor-alpha (TGF-alpha) increased 30-fold, while no increase in secreted TGF-alpha was detected. Furthermore, Ca2+-induced tyrosine phosphorylation of PKC-delta and phosphotyrosine-association of the receptor adapter protein Shc was diminished in EGFR -/- keratinocytes, suggesting that EGFR activation may occur during keratinocyte differentiation. Tyrosine phosphorylated PKC-delta was also detected in mouse epidermis, suggesting that this differentiation-associated signaling pathway is physiological. These results establish a requirement for the EGFR in Ca2+-induced tyrosine phosphorylation of PKC-delta, and document the production of cell-associated TGF-alpha in differentiated keratinocytes which may function independent of its usual mitogenic effects.

Production of infectious bovine papillomavirus from cloned viral DNA by using an organotypic raft/xenograft technique.

Bovine papillomavirus type 1 (BPV-1) induces fibropapillomas in its natural host and can transform fibroblasts in culture. The viral genome is maintained as an episome within fibroblasts, which has allowed extensive genetic analyses of the viral functions required for DNA replication, gene expression, and transformation. Much less is known about BPV-1 gene expression and replication in bovine epithelial cells because the study of the complete viral life cycle requires an experimental system capable of generating a fully differentiated stratified bovine epithelium. Using a combination of organotypic raft cultures and xenografts on nude mice, we have developed a system in which BPV-1 can replicate and produce infectious viral particles. Organotypic cultures were established with bovine keratinocytes plated on a collagen raft containing BPV-1-transformed fibroblasts. These keratinocytes were infected with virus particles isolated from a bovine wart or were transfected with cloned BPV-1 DNA. Several days after the rafts were lifted to the air interface, they were grafted on nude mice. After 6-8 weeks, large xenografts were produced that exhibited a hyperplastic and hyperkeratotic epithelium overlying a large dermal fibroma. These lesions were strikingly similar to a fibropapilloma caused by BPV-1 in the natural host. Amplified viral DNA and capsid antigens were detected in the suprabasal cells of the epithelium. Moreover, infectious virus particles could be isolated from these lesions and quantitated by a focus formation assay on mouse cells in culture. Interestingly, analysis of grafts produced with infected and uninfected fibroblasts indicated that the fibroma component was not required for productive infection or morphological changes characteristic of papillomavirus-infected epithelium. This system will be a powerful tool for the genetic analysis of the roles of the viral gene products in the complete viral life cycle.

The research on antioxidative properties of TOLPA Peat Preparation and its fractions.

The protective and therapeutic role of TPP and its fractions against lipid peroxidation in the mitochondria from human placenta as a model for experiments was evaluated. Both TPP and its fractions cause the decrease in MDA production. The antioxidant force of TPP and its fractions with antioxidant force of vitamin E was compared.