Objective: The first aim of the present study was to evaluate the effect of Echinacea purpurea - containing drug (Immunal forte tablets) on neovascular response induced in mice skin by human lung and kidney cancer cells, 3 days after their intracutaneous grafting. The second aim was to evaluate the effect of Immunal forte tablets, administered for seven days to healthy human volunteers, on the proportion of natural killer (NK) cells in their blood and on the activity of their blood granulocytes, tested by chemiluminescence test. Results: Echinacea preparation caused inhibition of angiogenesis induced by human lung and kidney cancer cells, as evaluated 3 days after intradermal cells injection. In human volunteers Immunal forte administration resulted in the stimulation of granulocytes metabolic activity and have increased the incidence of CD16(+) and CD56(+) NK cells in their blood.

Hyperglycaemia increases inflammatory cytokine concentration in the blood. Elevated levels of interleukin-18 (IL-18), a cytokine belonging to the interleukin-1 (IL-1) family, were recently reported in patients with Type 2 diabetes mellitus (DM2) and nephropathy. The aim of the present work was an examination of IL-18 concentration in the sera of elderly DM2 patients with nonproliferative retinopathy and age-matched control people and an estimation whether this cytokine plays pro- or anti-angiogenic role in in vivo angiogenic activity of their sera in mice cutaneous angiogenesis test. Recombinant human IL-18 injected intradermally to murine skin induced significant neovascular reaction. DM2 patients sera contained higher concentration of IL-18 and induced stronger neovascular reaction in mice skin than did the sera of corresponding control people. Sera from both groups of people after neutralization with antihuman IL-18 antibodies lost substantial part of their angiogenic activity.

Caffeine and its active derivative, theobromine, are probably the most frequently ingested pharmacologically active substances. Considering their uninhibited transport via the placental barrier as well as immature enzymatic activities and metabolic pathways in embryos and infants resulting in the longer half-life of methyloxanthines and their accumulation, unrestrained uptake of these substances might result in noticeably more pronounced biological effects during pregnancy and the postnatal period. Our previous studies have shown that methyloxanthines are significant inhibitors of angiogenic growth factors production and angiogenesis itself. We have hypothesized that increased uptake of these substances might affect embryonal angiogenesis and, later in the postnatal period, maturation and functional activity of the offspring's immune system. The study was performed on 2-month-old Balb/c mice fed theobromine 2 or 6 mg/day during pregnancy and lactation. On day 18 of pregnancy the number and weight of embryos were assessed as was their tissue angiogenic activity, using the cutaneous angiogenesis assay. In the group of 4-week-old sucklings, body and spleen were weighed together with the trunk, and tail and limb length were measured. Six weeks after birth the splenocytes' mitogen-induced activity and their ability to induce graft-versus-host reaction as well as the humoral response to SRBC antigen were evaluated. Content of theobromine in the embryos' tissue was estimated by high liquid performance chromatography (HPLC). Theobromine feeding resulted in significant inhibition of embryo growth as assessed by their weight and decreased angiogenic activity of their tissue. The theobromine content in embryo tissue from treated groups was higher than in the controls, and the difference was close to significant. In the postnatal period the discrepancies in the treated 4-week-old group's development were also observed in the significantly shorter limbs in comparison to the controls. Moreover in the treated group of 6-week-old sucklings, considerable variations in the immune system's functional activity were registered as far as cellular and immune response were concerned. Respectively, the splenocytes' mitogen-induced proliferative activity was significantly suppressed while the graft-versus-host reaction was up-regulated, and the serum antibodies titer was elevated in correspondence to the observed spleen enlargement. We concluded that a theobromine-enriched diet affects progeny development in both prenatal and postnatal periods. Consequently, particular attention should be paid to the reduction of theobromine consumption, and most probably that of other methyloxanthines, during pregnancy and lactation.

The immunomodulatory activity of acetylshikonin (ACS) and isobutyrylshikonin (IBS) was studied in female and male inbred Balb/c mice, and in F1 hybrids (Balb/c x C3H). ACS and IBS were isolated from Lithospermum canescens Lehm. (Boraginaceae) roots. Splenocytes from mice fed 40 microg of ACS had higher proliferative potential in cultures with PHA than corresponding controls and also higher migratory in vitro activity than splenocytes obtained from control animals. ACS at a 40 microg daily dose stimulated G-v-H reaction but inhibited it at a 200 microg dose. IBS at a 40 microg dose significantly increased humoral response.

Human serum injected intradermally into mouse skin induced strong neovascular response around the site of injection. Heating of serum for 30 min to 56(0) C significantly diminished the number of newly-formed blood vessels. No influence of heating on the levels of vascular endothelial growth factor, insulin-like growth factor, transforming growth factor beta and interleukin 18 were observed. Addition of epigallocatechin gallate, known as tknown thyrosine kinase inhibitor to human serum, gave the effect comparable to the effectthat of heating. Human serum-induced angiogenesis in the mouse skin test might be a valuable evaluation method assessing the effect of various compounds on the complement-dependent and complement-independent angiogenesis.

We have previously shown that some commonly consumed substances, present in the average human diet, might cross over the placental barrier and affect the development of embryo immune system. The aim of the present study was to examine the cellular and humoral immune response in the group of offsprings from mothers treated with caffeic acid (1 or 6 mg/day) during pregnancy and lactation. We observed that the mean spleen relative weight was reduced in 4-week old progeny of mice treated with 6 mg/day of caffeic acid. The percentage of spleen CD3 + lymphocytes was decreased in both examined doses, while the number of CD19 + lymphocytes was not affected. Local G-v-H reaction induced by splenocytes from 4-week old progeny of caffeic acid-treated mothers was significantly suppressed in both dosage groups. Caffeic acid feeding (6 mg) resulted in up-regulation of the peripheral blood granulocytes chemiluminescent activity as well as in the enhancement of anti-SRBC antibody production in the 6-week old progeny of the treated mothers. We concluded that caffeic acid-enriched mothers' diet may affect progeny immune system function in the postnatal period.

The effect of herbal remedy (further named HR), a complex preparation consisting of three substances: Echinacea purpurea extract, Allium sativum extract, and cocoa, on the growth of L-1 sarcoma, tumour angiogenesis, VEGF tumour concentration and Pseudomonas aeruginosa infection in mice were studied. A significant inhibitory effect of the remedy on tumour angiogenic activity using cutaneous angiogenesis test, and an inhibitory effect on L-1 sarcoma growth were observed. However, the concentration of VEGF was higher in tumours of mice treated with the remedy as compared to the control group. It was found that P. aeruginosa infection intensity was significantly lower (after treatment with the remedy) than in the control group. The possible mechanisms of action of the preparation and perspectives of its use in human clinic are discussed.

The influence of Eleuterococcus senticosus on cellular and humoral immune response was evaluated. The experiments were performed on animal models (Balb/c mice and F1 crossbreeds Balb/cxC3H). It was shown that Eleuterococcus has immunomodulatory properties. This substance enhanced the cellular response of the mouse immunological system (chemokinetic activity of mice spleen cells, GvH reaction). A stimulatory effect of Eleuterococcus on the humoral response (antibody production) was also observed. Eleuterococcus did not augment the angiogenic activity of human renal carcinoma cells.

UNLABELLED:We previously reported the inhibitory effect of various methyloxantines and phenolic compounds on tumor-induced angiogenesis and the production of angiogenic growth factors. The aim of the present work was to evaluate the effect of chocolate (CH), food containing substantial amounts of methyloxantine theobromine and polyphenols (mainly catechins), given to mice during pregnancy and the lactation period, on weight of organs, length of limbs, and bone vascular endothelial growth factor (VEGF) concentration (tested by ELISA), in 4-week old offspring. The study was performed on 2-month old Balb/c mice fed during pregnancy and lactation 400 mg of CH daily. Content of polyphenols (catechines) and theobromine in the chocolate was estimated by high liquid perforance chromatography (HPLC). Concentration of VEGF was tested by ELISA. Feeding pregnant mice chocolate produced the following effects: decrease of relative length of limbs and thigh bones in 4-week old progeny and decrease in VEGF content of offspring femoral bones.CONCLUSION:attention should be paid to possible unwanted effects of catechine- and methyloxantine-rich food and beverages during pregnancy and lactation. 200 mg of chocolate per mouse corresponds to 100 g per person.

Sharks have been claimed to be resistant to cancer and oil from their livers have been used in Scandinavian folk medicine as anti-tumor drug. Shark liver oil contains 40% or more of squalene. Fish liver oil is also rich in squalene and polyunsaturated n-3 fatty acids. The aim of this work was to determine the anti-angiogenic and anti-tumor effects of these substances, together with another Scandinavian traditional remedy--arctic birch ashes--in Balb/c mice after transplantation of syngeneic L-1 sarcoma. All substances tested, alone or in combinations, significantly diminished cutaneous angiogenesis induced by tumor cells, and tumor growth.

Aberrant neovascularization plays a crucial role in ocular complications in diabetic patients. Sera from these patients contain high levels of angiostimulatory factors, the most important of which is vascular endothelial growth factor (VEGF). Many authors have described elevation of angiotensin-converting enzyme (ACE) activity in the sera of diabetic patients. It is important to determine the possible relationship between these two phenomena. We studied ACE serum activity and VEGF concentrations in patients with type 1 and type 2 diabetes and retinopathy We also investigated the effect of their sera on cutaneous angiogenesis induced in mice by grafting healthy human mononuclear blood leukocytes. We found a negative correlation between the angiostimulatory effect and ACE level in the sera of patients with type 1 diabetes and no correlation between these two parameters in patients with type 2 diabetes. VEGF concentrations were lower and ACE activity was significantly higher in the sera of patients with type 1 diabetes than in the sera of those with type 2 diabetes.

Development of new blood vessels in solid tumors depends on changes in equilibrium between angiogenic stimulators and inhibitors. Overexpression of angiogenic growth factors has been shown in bladder carcinoma. The 'mice cutaneous angiogenesis test' is a good method for assessment of the total angiogenic potential of bladder cancer tissue. The analysis of the levels of proangiogenic factors could be useful for the choice of properly directed angiogenesis inhibitors. The aim of our study was to investigate the influence of blocking some angiogenic factors on the angiogenic activity of bladder cancer tissue. Tumor tissue obtained from 12 patients with invasive bladder carcinoma was used. Cancer tissue homogenates were incubated in the presence of specific antibodies against VEGF, bFGF, Il-8 and aFGF. Cytokine levels were determined using the ELISA test. Cutaneous angiogenesis assay in Balb/c mice was performed to detect the angiogenic activity of the tumor tissue. VEGF, bFGF and Il-8 were present in all examined cancer tissues (aFGF level was not estimated). Cytokine concentration and angiogenic activity of bladder cancer tissue showed individual variation. There was no correlation between the cytokines content in tumor tissue and the ability of this tissue to induce angiogenesis. Absorption caused significant reduction in cytokines level. The reduction of angiogenic activity was observed in the cancer tissue of 1 patient after VEGF absorption, in 3 patients' tissue homogenates after incubation with anti-aFGF and in 2 patients' homogenates after bFGF absorption. There was no reduction of angiogenic activity after Il-8 absorption.

The effect of shark liver oil on cutaneous angiogenesis induced in mice by intradermal grafting of tumour cells was evaluated. It was shown that this substance (Ecomer) suppressed neovascular response in mice grafted with sarcoma L-1 syngeneic cells, human kidney cancer and human urinary bladder cancer cells. In addition, strong stimulatory effect of this drug on mice blood granulocyte number and their metabolic activity was observed.

Preterm delivery is one of the greatest problems in obstetric care. One of the most commonly used treatments for high risk cases is salbutamol, a beta-2-adrenoceptor agonist. The aim of the present study was to determine if such treatment causes any changes in the neonatal immune system which should therefore be a concern in the care of the newborn. The experiments were performed on 4 to 5 or 6 to 7-week old female and male offspring of salbutamol-treated C3H/W inbred mice. In the first part of the study, the number of spleen cells, phenotypes and activity (phytohemagglutinin-induced proliferation, ability to induce local graft versus host reaction) were determined. We observed lowering of cell number and lowered proportions of cluster of differentiation (CD)3, CD4 and CD8 positive lymphocytes in spleens of progeny of salbutamol-treated mice. However, CD4+ to CD8+ ration was higher in the progeny of salbutamol-treated mothers than in the corresponding controls. In addition, reactivity to phytohemagglutinin and ability to induce local graft vs. host reaction were higher (popliteal lymph node test) or undisturbed (lymphocyte-induced angiogenesis test) in this group of mice.