Genetic diversity of urinary bladder cancer and the risk of recurrence based on mutation analysis.

The aim of the study was to assess the genetic diversity of bladder cancer and determine the suitability of a proposed molecular marker panel to monitor the course of bladder cancer patients. The study involved 185 patients with diagnosed bladder cancer. The genetic diversity of the bladder cancer was evaluated by the prevalence of mutations in the TP53, HRAS, FGFR3 and WWOX genes. Mutations were detected in 62.2% of the tumor samples. The most frequently mutated genes were FGFR3 (49.7%) and TP53 (16.2%). No mutation was observed in the WWOX gene. FGFR3 mutations, contrary to TP53, correlated with lower tumor stage and grade, and the presence of multiple tumors. The risk of death was significantly higher in patients with TP53 mutant tumors (HR=3.12; 95%CI: 1.14-7.27; p=0.006) but lower in patients with FGFR3 mutations (HR=0.36; 95%CI: 0.15-0.87; p=0.002). None of the investigated genes was an independent predictor of disease-specific survival, recurrence-free survival or progression-free survival. The results confirm the existence of two alternative pathways of bladder cancer. However the presence of a high percentage of wild type variants in the higher stages of the disease suggest the existence of another pathway of molecular changes leading to the development of bladder cancer. Molecular analysis may have prognostic value and may facilitate the assignment of patients to appropriate forms of treatment - especially in the case of patients with a T1 tumor, where different mutational patterns were observed in each grade.

MMP, VEGF and TIMP as prognostic factors in recurring bladder cancer.

OBJECTIVES: To investigate the clinical correlates and prognostic utility of MMP, VEGF and TIMP genes expression in bladder cancer (BCa) recurrence. METHODS: Expression of MMP1, MMP2, MMP9, VEGFA and TIMP1, TIMP3 was analyzed by qRT-PCR using SYBR Green in peripheral blood leukocytes (PBLs) of BCa patients at two time points (diagnosis (n=40), and first recurrence (n=40)) and an age-matched group of healthy controls (n=100). Plasma concentrations of MMP1 (pro- and active forms) were measured using ELISA in BCa patients. RESULTS: The expression of MMP1 mRNA was significantly lower in BCa patients with first recurrence compared to control (p=0.019). Expression of other genes did not differ significantly between the groups. MMP9 gene expression was associated with differentiation grade (p=0.043), with the highest expression in poorly differentiated tumors (G3) and was higher in smokers than in non-smokers (p=0.039) in BCa patients at diagnosis. The results at two time points showed that MMP9 and VEGFA genes expression was increased in patients with moderately differentiated BCa (p=0.029), and advanced pathologic stage (p=0.048), respectively. Moreover, gene expression of TIMP1 was increased for G3 (p=0.043), and was decreased for early recurrence (p=0.003). CONCLUSIONS: Our study suggests that the expression of MMP9 in PBLs of BCa patients at diagnosis is associated with the differentiation grade of the BCa, and smoking status. Genes expression of MMP9, VEGFA and TIMP1 in PBLs may play a pivotal role in regulation of progression of BCa. Additionally, TIMP1 gene expression may be important factor for early recurrence of BCa.

Expression of NRF2 and NRF2-modulated genes in peripheral blood leukocytes of bladder cancer males.

Nuclear factor (erythroid-derived 2)-like 2 (NRF2) is an oxidant-responsive transcription factor involved in induction of antioxidant genes. We assessed NRF2 and selected NRF2-modulated gene expression: glutathione S-transferase A1 and P1 (GSTA1 and GSTP1), mitochondrial superoxide dismutase (SOD2) in blood leukocytes of 51 bladder cancer patients and 90 control males. A significant up-regulation of SOD2 expression (P=0.002) was observed in leukocytes of patients. NRF2 expression was positively correlated with GSTP1 and with SOD2 mRNA level, both in patients and controls. These data suggest disturbances in SOD2 transcription in circulating blood leukocytes of males with bladder cancer. Moreover, concomitant constitutive expression of NRF2 and its target genes may suggest important role of NRF2 transcription factor in positive regulation of antioxidant genes, resulted in enhanced cytoprotection in human peripheral blood leukocytes.

Influence of the pH of the cultivation medium on survival and development of stage III larvae of Anisakis simplex.

The cultivation was done on Eagle's media as well as Eagle's media enriched with the supplement of 10 and 20% of bovine serum. In the media without serum the larvae lived significantly shorter and did not get through molting. They lived the longest and came through molting the fastest in the media with pH 2.0 supplemented with 20% SB. In the media with the pH 4.0 and 7.3 the larvae lived significantly shorter and only a small proportion reached stage IV.

[The influence of various levels of carotene and protein in the diet on the index of invasion and digestive enzyme activity in chickens infected with Ascaridia galli]. / Wplyw zróznicowanego poziomu karotenu i bialka w diecie na wskazniki inwazji oraz aktywnosc enzymów trawiennych u kurczat zarazonych Ascaridia galli.

Chickens 10 days old were infected with 500 eggs of Ascaridia galli. They were fed with diet containing 11% or 19% protein. Some of them were given beta-carotene in a dose equivalent to daily requirement of vitamin A, others were given double dose of beta-carotene. The intensity and extensity of invasion, the body weight, and activities of alpha-amylase, lipase and trypsin in pancreas and duodenal content were measured. Application of beta-carotene (especially in a double dose) caused the decrease in the invasion index and moderated the changes in the activity of digestive enzymes, occurring in the infected chickens.

[Activity of alpha-amylase, trypsin and lipase in pancreas and duodenal contents of Ascaridia galli infected chickens fed with various proteins]. / Aktywnosc alfa-amylazy, trypsyny i lipazy w trzustce i tresci dwunastniczej kurczat zarazonych Ascaridia galli zywionych pasza o róznej zawartosci bialka.

The study was carried out on Astra S chickens which were grown on diets containing 11% and 19% proteins. In homogenized pancreas and duodenal contents from control animals and chickens infected with 500 invasive eggs of Ascaridia galli activities of alpha-amylase (Fennel method), lipase (Cherry-Crandall method) and trypsin (Anson method) were determined. After 7 weeks of the invasion the activities of these enzymes were higher in duodenal contents and lower in pancreas of infected birds in comparison with the control animals. The differences were significant for alpha-amylase and lipase activities in animals which were given 11% protein diet, and for trypsin activity of chickens groups fed with 19% protein diet.

[Effect of Hydrochloric acid on invasion of Ascaris suum; enzyme activity in the digestive system and serum of newborn piglets]. / Wplyw roztworu kwasu solnego na inwazje ascaris suum oraz aktywnosc enzymów trawiennych przewodu pokarmowego i surowicy prosiat osesków.

The studies were carried out on twenty newborn piglets. They were divided into four groups. The groups no 3 and no 4 were given intragastric 0.18% HCl from the 3th day of experiment. The groups no 2 and no 4 were infected on the 7th day with 10,000 invasive eggs of Ascaris suum. The presence of A. suum larvae in the lungs and liver was examined after one week lasting invasion by Baermann method. The total acidity in the gastric content was measured. The activity of alpha-amylase, lipase and proteases was determined in the extracts from pancreas and in the contents of stomach, duodenum and jejunum. The level of pepsinogens and alpha-amylase in the animals serum was studied. The intensity of Ascaris invasion was slightly higher in the group which was given HCl than in the infected group without HCl. The activity of digestive enzymes in the both groups was similar. Only in the stomach content from the 4th group the activity of pepsin was higher (p < 0.05), and alpha-amylase and lipase were lower (p < 0.01) than in the 2nd and in the 3th group. The level of pepsinogens was always lower and alpha-amylase higher in the serum of infected animals than in uninfected groups.

[Effect of vitamin A and protein levels in the diet of chickens infected with Ascaridia galli on activity of digestive enzymes in pancreas and small intestine]. / Wplyw poziomu witaminy a i bialka w diecie kurczat zarazonych nicieniem ascaridia galli na aktywnosc enzymów trawiennych w trzustce i jelicie cienkim.

Hybrid chickens Astra S fed vitamin A deficiency diet containing 11 or 19% protein were infected with 500 invasive eggs of Ascaridia galli. One part of them received vitamin A in a dose corresponding to daily requirement, other part in double dose. After 7 weeks the extensity and intensity of ascaridiosis were determined. In pancreas and intestinal contents the activities of alpha-amylase, lipase and trypsin were measured. The invasion index was the highest in the group of chickens fed the protein and vitamin A deficiency diet. Vitamin A in a requirement dose lowered the parasite index. The results with double dose of vitamin A were worse in comparison with single dose. The activities of digestive enzymes were lower in infected animals than in the control groups. Vitamin A acted as moderator of the enzymatic changes during ascaridiosis.

[Processes connected with hatching of larvae from eggs of parasitic nematodes]. / Procesy zwiazane z wyleganiem sie z jaj larw nicieni pasozytniczych.

Influence of such factors as temperature, pH, CO2 level and oxidation- reduction potential value in the alimentary tract of host on the stimulation of invasive eggs was studied. Changes of these factors lead to change in permeability of inner layer of egg-shell. Then the hatching fluid is released and its enzymes digest the egg-shell. In the nematodes, which have the free-living larvae, the initiation process can depend on parasite or environmental agents.

[The influence of HCl administration on the acidity and the activity of enzymes in the stomach contents of suckling piglets]. / Wplyw podawania HCl na kwasowosc i aktywnosc enzymów w tresci zoladka nie odchowanych prosiat.

After HCl administration to 3-21 day old crossbred piglets (wbp x pbz x zlotnicka biala) the total acidity and output of acidity, the activity of pepsin, alpha-amylase and lipase were measured in the stomach content. The activity of pepsinogen was estimated in piglet's serum. The results do not indicate the significant influence of HCl on the acidity and the activity of enzymes in the stomach content.

[The influence of HCl administration on the activity of trypsin, alpha-amylase and lipase in different parts of the alimentary tract of piglets in the first three weeks of life]. / Wplyw podawania HCl na aktywnosc trypsyny, alfa-amylazy i lipazy w róznych odcinkach przewodu pokarmowego prosiat w pierwszych 3 tygodniach zycia.

The experiments were conducted using 3 to 21 days old piglets. The content of animals stomach was supplemented with 0.18% HCl starting from the 3rd, 7th and 10th day of their life. The animals were dissected on the 4th, 7th and 11th day after HCl administration. The activity of trypsin, alpha-amylase and lipase was measured in homogenate from pancreas and duodenum and ileum contents. The presented results do not confirm the existence of significant differences between control and experimental animals.

[The influence of an infective dose and the duration of infection with Ascaridia galli on trypsin and alpha-amylase activity in the pancreas of chickens fed with diets of different protein levels]. / Wplyw dawki zarazeniowej i trwania inwazji Ascaridia galli na aktywnosc trypsyny i alfa-amylazy w trzustce kurczat zywionych pasza o zróznicowanym poziomie bialka.

The studies were carried out on the chickens given the diet containing 19% and 11% of protein. On the 7th day of their life one part of the chickens were infected with different doses (125-500) of invasive eggs of Ascaridia galli. The other part of the birds were infected with 500 eggs of A. galli and the invasion lasted from 5 to 49 days. In the extracts from pancreas the activities of trypsin and alpha-amylase were measured. No differences were observed between trypsin activity of control and infected chickens fed with diet containing 11% of protein. The activity of trypsin was lower in infected chickens fed with diet containing 19% of protein, when the invasion lasted over 15 days. The activity of alpha-amylase in both groups decreased after infection with more than 100 invasive eggs of A. galli. The intensity and the extensiveness of invasion were lower in the chickens given the diet of 19% of protein.

[Effects of administration of vitamin A, B 2 and B 15 to guinea pigs during the larval stage of ascariasis on alpha-amylase activity in the lungs and kidneys]. / Wplyw witamin A, B2 i B15 podawanych swinkom morskim w czasie glistnicy larwalnej na aktywnosc alfa-amylazy w plucach i nerkach.

The guinea pigs were administrated vit. A (400 i.u.), vit. B2 (1 mg) or vit. B15 (5 mg). On the 9-th day of the experiment part of them was infected with 5000 invasive eggs of Ascaris suum. The invasion lasted 6 days and was controlled by lungs and kidney weight, and number of larvae in the lungs. The activity of amylase was determined by saccharogenic method in both organs. In the lungs of infected animals the activity of alpha-amylase was about 3 times lower than in the control. The infection of guinea pigs which were given vitamins did not cause change of enzyme's activity. In the kidney directive tendency was the same, but the differences were smaller. The infection resulted in an increase of relative mass of lungs. This index and number of larvae was considerable smaller in guinea pigs with vitamins A and B15 administration. Any testing agent did not cause change of relative weight of kidney.

[Trypsin and alpha-amylase activity in the pancreas of guinea pigs. V. Effects of administration of vitamin A and B 2 during the larval stage of ascariasis]. / Aktywnosc trypsyny i alfa-amylazy w trzustce swinek morskich. V. Wplyw podawania witamin A i B2 w czasie askarydozy larwalnej.

The infection of guinea pigs with Ascaris suum larvae resulted in decrease of the activities of trypsin and alpha-amylase, and in increase of lipase activity in extracts from their pancreas. The activity of alpha-amylase, lipase and the relative weight of lungs of infected animals which were given vitamin A, did not differ from control animals. The activity of trypsin from pancreas these animals was higher than that measured in only infected guinea pigs but it was lower than in control animals. Application of vitamin B2 and the infection of guinea pigs with A. suum did not lead to the synonymous results.

[Alpha-amylase activity in serum and some organs of guinea pigs with larval ascariasis]. / Aktywnosc alfa-amylazy w surowicy i niektórych narzadach swinek morskich podczas askarydozy larwalnej.

The studies were carried out on 40 guinea pig males weighting about 230 g. The experimental animals were infected with 7000 invasive eggs of Ascaris suum. On the 3-rd, 7-th and 10-th day after infection the activity of alpha-amylase was estimated according to Caraway's method in serum, liver, pancreas, lungs, kidneys and spleen. The infection of guinea pigs results in increased activity of enzyme in serum and spleen, and decreased in pancreas, liver, kidneys and lungs. On the 3-rd day after infection the changes in amylase activity were the most intense.

[Trypsin and alpha-amylase activity in the pancreas of guinea pigs. I. The influence of infective dose and duration of infection by Ascaris suum Goeze, 1782]. / Aktywnosc trypsyny i alfa-amylazy w trzustce swinek morskich. I. Wplyw wielkosci dawki zarazeniowej i czasu trwania inwazji Ascaris suum Goeze, 1782.

The studies were carried out on guinea pig males weighting about 280 g. The animals were infected with 5000 or 20,000 invasive eggs of Ascaris suum. On the 3-rd, 7-th, 10-th and 14-th day of invasion the animals were dissected. Lungs were weighted, and the number of Ascaris larvae was estimated using Baermann method. The activity of alpha-amylase (Fennel's method) and trypsin (Anson's method) were examined in pancreas homogenate. The infection of guinea pigs with A. suum larvae resulted in a decrease of amylase and trypsin activity in pancreas. The activities were the lowest at the 3-rd day of invasion for amylase and at the 7-th day for trypsin. The results obtained for the animals infected 5 or 20,000 invasive eggs A. suum did not differ statically.