The anticancer activity of alpha-tomatine against mammary adenocarcinoma in mice.

AIM: To evaluate the anticancer effect of alpha-tomatine (i.p.) either alone or in combination with doxorubicin (i.v.) in a mouse tumour model. METHODS: We studied the effect of repeated alpha-tomatine (0.1 - 9 mg/kg) and/or doxorubicin (2 mg/kg) on the growth and mitotic activity of the solid Ehrlich tumour in vivo, as well as on the survival of the tumour-bearing mice. RESULTS: Monotherapy with alpha-tomatine had a significant dose-dependent anticancer effect which peaked at 1 mg/kg. This was shown by both slowed tumour growth and reduced tumour cell proliferation. We also provide the first evidence that the combination alpha-tomatine (1 mg/kg) and doxorubicin (2 mg/kg) had a synergistic effect and significantly prolonged the survival of the mice. Neither alpha-tomatine nor doxorubicin influenced the infiltration of tumours with CD3+ lymphocytes; nor were we able to find an in vivo modulation of the key molecules of two regulatory pathways reported in vitro as the principal anti-cancer mechanisms of alpha-tomatine, i.e. iNOS and phosphorylated ERK2. However, alpha-tomatine still led to intracellular DNA inhibition and protein synthesis in Ehrlich tumour cells in a short-term culture ex vivo with IC50 values of 8.7 and 6.6 µM. CONCLUSIONS: The results suggest that ΤΟΜ, especially in combination with doxorubicin, may be a promising agent for the treatment of malignant solid tumours. Despite growing knowledge of the mechanisms of ΤΟΜ action in cancer cells, most aspects remain unclear. Parallel organ toxicity, especially potential liver effects, requires careful attention when performing in vivo studies in the future.

Total HIP arthroplasty in patients with Parkinson's disease.

The aim of this study is to evaluate the results of total hip arthroplasty in patients with Parkinson's disease during a period of five years, focusing on the assessment of the risks and benefits of surgery. During this period we performed total hip arthroplasty in 14 patients (15 hips) with Parkinson's disease. Patients were evaluated by subjective symptoms and objective findings, with a focus on the use of support while walking and walking distance, severity of Parkinson's disease before surgery and at the time of the last follow-up. During the postoperative period, the following parameters were assessed: length of ICU stay, mobilization, complications, the total duration of hospitalization and follow-up care after discharge. Of the 11 patients (12 hips) followed-up 1-5 years with an average of 3 years after operation 8 cases showed progression of neurological disability. 5 patients (6 hips) showed an increased dependence on the use of support when walking and reduced distance that the patient was able to walk. Subjectively, 10 hip joints were completely painless and 2 patients complained of only occasional mild pain in the operated hip. Complications that were encountered were urinary tract infection (5 patients), cognitive impairment (3 patients) and pressure ulcer (2 patients). We did not observe any infection or dislocation of the prosthesis. Three patients fell and fractured the femur and 3 patients in our cohort died during follow up. Implantation of total replacement is possible with judicious indication after careful evaluation of neurological finding in patients with minimal or mild functional impairment of the locomotor system. Prerequisite for a good result is precise surgical technique and optimal implant position with balanced tension of the muscles and other soft tissues around the hip.

Differences in vanadocene dichloride and cisplatin effect on MOLT-4 leukemia and human peripheral blood mononuclear cells.

Modern chemotherapy is interested in developing new agents with high efficiency of treatment in low-dose medication strategies, lower side toxicity and stronger specificity to the tumor cells. Vanadocene dichloride (VDC) belongs to the group of the most promising metallocene antitumor agents; however, its mechanism of action and cytotoxicity profile are not fully understood. In this paper we assess cytotoxic effects of VDC in comparison to cisplatin using opposite prototype of cells; human peripheral blood mononuclear (PBMCs) cells and human acute lymphoblastic leukemia cell line (MOLT-4). Our findings showed cytotoxic effect of VDC on leukemia cells, but unfortunately on human peripheral blood mononuclear cells as well. VDC induces apoptosis in leukemia cells; the induction is, however, lower than that of cisplatin, and in contrary to cisplatin, VDC does not induce p53 up-regulation. Cytotoxic effect of VDC on leukemia cells is less pronounced than that of cisplatin and more pronounced in PBMCs than in MOLT-4 cells.

Evaluation of skin microcirculation during hemodialysis.

BACKGROUND: Cardiovascular mortality in hemodialysis (HD) patients remains high despite improvements in HD technique such as dialysis adequacy, dialysis fluid purity, and membrane biocompatibility. Optimal fluid balance to maintain optimal hemodynamic stability during hemodialysis (HD) procedure is essential. At the present time, hemodynamic stability is conventionally assessed as stability of macrocirculation, especially as maintenance of systemic blood pressure with no attention paid to peripheral perfusion. Peripheral ischemic vascular disease represents a serious problem with high mortality and morbidity. We estimated skin microcirculation changes during HD using new device, Laser Doppler Line Scanner (Moor Instruments, Devon, UK). AIMS: The aims were to introduce the novel method of detection of skin perfusion changes during hemodialysis and to evaluate possible relationship of these to ultrafiltration as well as to selected biochemical characteristics. METHODS: In 36 hemodynamically stable patients, we performed paired measurements of skin blood flow in both hands before and during HD with registering the time of dialysis and the total ultrafiltration achieved. RESULTS: We found a significant decrease in a majority of the evaluated areas. However, the skin blood flow change was not homogenous as it decreased more on the fingers. CONCLUSION: To our knowledge, this is the first study when the microcirculatory changes during hemodialysis are demonstrated and evaluated in large skin surface area, and showing not only a decrease in a majority of areas but also the heterogeneity of the changes.

Evaluation of the antineoplastic activity of L-rhamnose in vitro. A comparison with 2-deoxyglucose.

The effect of unsubstituted deoxyhexoses, 2-deoxy-D-glucose (2-DG) and L-fucose, on tumor cells has been reported in several papers throughout the last decades. That of a similar deoxysugar, L-rhamnose, which is synthesized in bacteria and plants but not in animal cells, has until today not been explored. In the present study, we examined the effect of L-rhamnose on DNA and protein synthesis, growth and the potential induction of apoptosis of tumor cells in vitro. Using 2-DG for comparison, we studied the effect of L-rhamnose in concentrations up to 20 (32 resp.) mmol/l on the initial velocity of the incorporation of labeled precursors of DNA and proteins in short term cultures of both mouse Ehrlich ascites tumor (EAT) and human HL-60 cells in vitro, and further, on cell proliferation and apoptosis induction in HL-60 cells. Neither cytotoxic nor cytostatic effects of L-rhamnose were observed with the exception of slightly pronounced inhibition of DNA synthesis in EAT cells. From the lacking inhibition of the protein synthesis it can be considered that L-rhamnose does not interfere with energy metabolism, at least not in a similar manner as 2-DG.

Increased melibiose/rhamnose ratio in bile of rats with acute cholestasis.

BACKGROUND AND AIM: Melibiose/rhamnose permeability test is used for noninvasive intestinal mucosa barrier testing. However, the possible escape route of the absorbed saccharides through either intact or impaired blood-biliary barriers has not so far been explored. The objective of the present study was therefore two-fold: First, to describe in detail the biliary pharmacokinetics of melibiose and rhamnose in rats; second, to evaluate the changes of both sugars' pharmacokinetics upon impairment of the blood-biliary barrier by acute extrahepatic cholestasis in rats. METHODS: Bile duct obstructed (BDO), sham-operated and intact (unoperated) male Wistar rats were administered, 24 h after the appropriate intervention, with a single intravenous dose of melibiose and rhamnose, and a 4-h pharmacokinetic study was performed. RESULTS: In intact animals, the biliary excretion of melibiose and rhamnose was only 0.06% and 0.4% of the administered dose, respectively, while the urinary excretion accounted for 70.6% and 61.7%, respectively. In BDO animals, the biliary excretion rate of both saccharides, especially that of melibiose, was increased with a consequent 4.4-fold rise of the biliary melibiose/rhamnose ratio, the accepted paracellular permeability indicator. Both, the renal clearance of melibiose and the urinary melibiose/rhamnose ratio remained uninfluenced by cholestasis. CONCLUSION: The present study is the first to describe in detail pharmacokinetic parameters and the biliary excretion of melibiose and rhamnose in healthy and cholestatic rats. The altered melibiose/rhamnose biliary excretion ratio in BDO rats indicates that the test is able to detect the impairment of the blood-biliary barrier in acute extrahepatic cholestasis.

Effect of atorvastatin on soluble CD14, CD40 Ligand, sE- and sP-selectins and MCP-1 in patients with type 2 diabetes mellitus: relationship to cholesterol turnover.

Metabolic abnormalities frequently associated with type 2 diabetes, feature besides endothelial dysfunction a novel factor of low cholesterol absorption and high cholesterol synthesis. We hypothesized an association between endothelial dysfunction and disturbances in cholesterol turnover, predisposing advanced atherosclerosis in patients with type 2 diabetes. We studied 75 patients: 30 with type 2 diabetes, 30 non-diabetic subjects with a history of cardiovascular disease, and 15 healthy subjects. Plasma sterols, soluble adhesion molecules sCD14, sCD40 Ligand, monocyte chemo-attractant protein-1 (MCP-1), sE- and sP-selectins were measured with and without atorvastatin therapy. The diabetic patients showed significantly higher levels of lathosterol and lower levels of sitosterol and campesterol. Non-diabetic subjects showed no significant differences in non-cholesterol based sterols. Plasma levels of hsCRP, sE- and sP-selectins and MCP-1 were significantly increased in patients with diabetes. The plasma levels of sCD40L correlate significantly with clinical parameters of BMI and glycaemia, and the plasma levels of sP-selectin correlate significantly with parameters of glycaemia and HbA(1c). The diabetic patients without statin therapy showed a significant correlation of sE-selectin with the marker of cholesterol absorption-sitosterol and sitosterol/cholesterol ratio. The diabetic patients without statin therapy showed a significant inverse correlation of sP-selectin with the marker of cholesterol synthesis-lathosterol. Both relationships disappeared with statin treatment. We conclude that endothelial dysfunction in obese patients with type 2 diabetes mellitus and cardiovascular disease associates with obesity and glycaemic control. The relation of parameters of endothelial dysfunction (such as sP-selectin and sE-selectin) with cholesterol synthesis and absorption may be influenced by reverse cholesterol transport.

The importance of abrogation of G2-phase arrest in combined effect of TRAIL and ionizing radiation.

BACKGROUND: In this work we studied the relationship between the enhanced expression of DR5 receptor and the effect of combination of TRAIL and ionizing radiation on cell cycle arrest and apoptosis induction in human leukemia cell line HL-60. MATERIAL AND METHODS: DR5, APO2.7 and cell cycle were analyzed by flow cytometry. Proteins Bid and Mcl-1 were analyzed by Western-blotting. For clonogenic survival, colony assay on methylcellulose was used. RESULTS: Ionizing radiation caused significantly enhanced positivity of DR5 receptors 24 h after irradiation with high doses (6 and 8 Gy). An increase of DR5 receptor positivity after a dose of 2 Gy was not statistically significant and application of TRAIL 48 h after irradiation did not increase the apoptosis induction. However, a decrease of radiation-induced G(2) phase arrest and an increase of apoptosis were observed when TRAIL was applied 16 h before irradiation with the dose of 2 Gy. Incubation with 6 microg/l TRAIL for 16 h reduced D(0) value from 2.9 Gy to 1.5 Gy. The induction of apoptosis by TRAIL was accompanied by Bid cleavage and a decrease of antiapoptotic Mcl-1 16 h after incubation with TRAIL. CONCLUSION: TRAIL in concentration of 6 microg/l applied 16 h before irradiation by the dose of 1.5 Gy caused the death of 63% of clonogenic tumor cells, similarly as the dose of 2.9 Gy alone, which is in good correlation with the enhanced apoptosis induction.

[MIAPS--programme system for order estimation of mutual similarity/dissimilarity of proteins behavior in time]. / MIAPS--programový systém pro odhadování poradí vzájemné podobnosti/nepodobnosti chování proteinu v case.

Statistical order estimation of mutual similarity/dissimilarity of proteins' behavior in time is the main purpose of the programme system MIAPS. Comparing with Correspondence Analysis (CA) method, mostly used so far, is simultaneously performed. Time series of 2-DE maps are requested as the data. MIAPS is available on http://www.lfhk.cuni.cz/knizekj/ and http://www.utia.cas.cz/user_data/sindelar/miaps.htm.

Contribution of phototherapy to the treatment of episiotomies.

OBJECTIVE: The purpose of this study was an objective consideration of possible benefits of phototherapy implemented with therapeutic laser or possibly polarized light in treating episiotomy, which is the most frequent obstetric intervention. MATERIALS AND METHODS: In the present study, the authors treated a total of 2,436 women. The light sources were as follows: a laser of a wave length 670 nm, power 20 mW, with continuous alternations of frequencies 10 Hz, 25 Hz, and 50 Hz, a polarized light source of a 400-2,000 nm wavelength in an interval of power 20 mW and frequency 100 Hz and a monochromatic light source of a 660 nm wavelength and power 40 mW, with simultaneous application of a magnetic field at an induction 8 mT. RESULTS: The work demonstrated high healing effects with minimum secondary complications in the treatment of episiotomies using a therapeutic laser at an energy density of 2 J/cm(2). The application of polarized light at an energy density of 5 J/cm(2) also exerted favorable therapeutic effects.