Biomedical subjects

B Falkiewicz

Publications and source records attributed to B Falkiewicz.

At least 19 recordsLinked to original sources

PubMed2002

p53 Gene status in relation to ex vivo chemosensitivity of non-small cell lung cancer.

PURPOSE: About 40% of non-small cell lung cancer (NSCLC) patients are candidates for systemic chemotherapy, despite the fact that at diagnosis most NSCLC are usually chemoresistant both in vivo and ex vivo. It is important to develop sufficient methods of prediction of the response to chemotherapy and to find molecular markers that may prognose this response. Therefore, a study on the relationship of p53gene status to the ex vivo chemosensitivity of primary human NSCLC was performed. METHODS: Three drug combinations (carboplatin/etoposide, cyclophosphamide/etoposide/epirubicin, and paclitaxel/carboplatin) were tested in a modified ATP cell viability assay. A group of 28 cases of primary human NSCLC was assessed. RESULTS: Ex vivo chemosensitivity testing showed that tumors with p53 mutations were significantly more resistant to the cyclophosphamide/etoposide/epirubicin regimen than with normal p53 gene ( P = 0.012). However, no correlation was observed for two other treatment regimens. CONCLUSION: Mutations in the p53gene can lead to enhanced chemoresistance, confirming the hypothesis that the p53 gene may serve as a marker of tumor response to treatment in NSCLC. However, the data also illustrate that some additional factors might contribute to drug resistance of the examined tumors.

Aged↗

PubMed2001

The suitability of DNA extracted from formalin-fixed, paraffin-embedded tissues for double differential polymerase chain reaction analysis.

Formalin-fixed, paraffin-embedded (FFPE) tissues are one of the popular sources of diagnostic materials, the easiest to store and transport. They are often used as the source of nucleic acids for retrospective molecular analyses based on DNA amplification by polymerase chain reaction (PCR). However, it is known that nucleic acids from paraffin-embedded tissues are much worse templates than those recovered from fresh tissues. It is exceptionally important in a quantitative analysis, including double differential PCR (ddPCR). Therefore, a pilot study comparing quantity and quality of DNA extracted with various paraffin removal and DNA isolation procedures from FFPE tissues was conducted. Furthermore, the suitability of DNA isolated with optimized procedure for the assessment of erbB-2 average gene copy number (AGCN) was checked. Specimens for comparison of extraction and isolation procedures were generated from the same human normal thyroid tissue embedded in paraffin to eliminate variabilities in tissue processing and sample size. Three procedures of paraffin removal and three procedures of DNA extraction from deparaffinized tissue were compared. Only one procedure provided DNA, which was efficiently amplified in ddPCR. The material obtained with this optimized procedure was used to check the precision of ddPCR by evaluation of AGCN of erbB-2 oncogene. Low variability of obtained results close to expected AGCN value (AGCN=1) indicates high reproducibility of the method, as well as its high accuracy, if the normal value of erbB-2 AGCN in the examined tissue is assumed.

DNA↗

PubMed2001

Porphyria cutanea tarda, hepatitis C virus and selected symptoms of cholestasis.

BACKGROUND: The influence of hepatitis C virus (HCV) infection on the development of porphyria cutanea tarda (PCT) was recently detected, but the status of trace elements in etiology of the diseases is relatively poorly recognized. Individually observed symptoms of cholestasis do not seem to be related to the above conditions. MATERIAL AND METHODS: A group of 22 PCT patients (37-73 years old, mean 54.9 years), including 10 cases (45.5%) of serum HCV-RNA positive, as evaluated by automatic Cobas AmplicorTM Hepatitis C ver. 2.0 assay (Roche Diagnostics), was assessed. None of the patients have recently abused alcohol. Serum levels of copper and bilirubin, activity of gammaglutamyltranspeptidase (GGTP) and alkaline phosphatase (ALP) were measured in all patients. RESULTS: We have found statistically significant differences between both mean serum activity of GGTP (145.9 U/l vs. 58.3 U/l, p = 0.004) and mean serum copper levels (131.9 mg/dl vs. 79 mg/dl, p = 0.003) in patients infected with HCV compared to the other PCT patients, respectively. Mean serum ALP activity was also higher, but the difference was not of statistical importance (106.9 U/l vs. 90.9 U/l, p = 0.24); mean serum bilirubin concentration was significantly lower in HCV-infected patients (0.55 mg/dl vs. 0.85 mg/dl, respectively, p = 0.035). CONCLUSION: In our group of PCT patients we have noticed a higher serum copper level and higher activity of both GGTP and alkaline phosphatase, but lower level of serum bilirubin in patients infected with HCV as compared with those not infected. This observation does not confirm suggestions concerning the role of HCV in promotion of cholestatic liver disease in PCT patients. The influence of HCV infection on the metabolism of copper and GGTP activity is connected rather with the inflammatory lesions in the liver of PCT patients infected with HCV. Bilirubin metabolism in PCT patients requires further studies for its elucidation.

Adult↗

PubMed2001

Influence of hepatitis C virus (HCV) infection on porphyrin and iron metabolism in porphyria cutanea tarda (PCT) patients.

BACKGROUND: Porphyria Cutanea Tarda (PCT) is the most common form of porphyria. It is characterised by lowered activity of uroporphyrinogen decarboxylase. It seems possible that the hepatitis C virus (HCV) infection triggers the symptoms of PCT. MATERIAL AND METHODS: A group of 29 PCT patients (33-73 years old, mean 54 years) was evaluated. In these patients the serum HCV-RNA (by mean of automatic Cobas AmplicorTM Hepatitis C ver. 2.0 assay, Roche Diagnostics), activity of transaminases (ALT, AST), serum protein fractions, iron, total iron binding capacity (TIBC), ferritin, transferrin, transferrin saturation, and urinary porphyrins excretion were assessed. RESULTS: The presence of HCV-RNA was detected in 12 cases (41.4%). Statistically important differences between currently HCV infected patients and other PCT patients concerned the following parameters: mean body mass (68.5 vs. 78.0 kg, p = 0.02), mean body mass index (22.4 vs. 26.8 kg/m2, p = 0.007), mean serum activities of ALT (112.6 vs. 65.4 U/l, p = 0.02) and AST (85.0 vs. 56.5 U/l, p = 0.038), total serum protein concentration (78.0 vs. 71.6 g/l, p = 0.016) and serum alpha-2 protein concentration (7.2 vs. 5.5 g/l, p = 0.009), respectively. Some other not significant differences were also found: mean serum iron level 182 vs. 169 micrograms/dl, transferrin concentration 2.77 vs. 2.56 g/l, transferrin saturation 43 vs. 45%, serum TIBC 445 vs. 397 mg/dl and ferritin concentration 394 vs. 400 micrograms/l, respectively. The mean urinary uroporphyrin concentrations were 643 micrograms/l vs. 366 micrograms/l (p = 0.16) and those of coproporphyrins were 190 micrograms/l vs. 136 micrograms/l (p = 0.37), respectively. CONCLUSION: In the group of PCT patients investigated no significant relationship between current HCV infection and iron or porphyrin metabolism was noted. Nevertheless, HCV infection significantly influences the symptoms of liver damage and seems to influence other metabolic parameters in PCT patients.

Adult↗

PubMed2001

HCV-RNA detection in liver bioptates--a comparison of automatic and 'home-made' protocols combined with a new procedure of HCV-RNA extraction.

BACKGROUND: For HCV-RNA detection in liver tissue a generally accepted reference method has not been established yet. Therefore, we have developed a procedure of HCV-RNA extraction from liver tissue and compared two methods of HCV-RNA detection. MATERIALS AND METHODS: A set of 32 liver biopsy specimens, obtained from chronically HCV infected patients, has been examined. At the time of biopsy, serum HCV-RNA was detectable in 20 patients in RT-PCR automatic Cobas AmplicorTM Hepatitis C assay, ver. 2.0 (Roche Molecular Systems, Inc, Pleasanton, CA, USA) 2 serum samples were negative for HCV-RNA and 10 patients has not been tested. Liver tissue has been homogenized in the presence of CRSR-Green (Fast RNA Kit-Green, Bio101, Inc, Vista, CA, USA) and a mixture of phenol/chloroform/isoamylic alcohol, using a FastPrep homogenizator (Bio101, Inc, Vista, CA, USA). Then RNA has been isolated from the material obtained using the Total RNA Prep Plus procedure (A&A Biotechnology, Gdańsk, Poland). Presence of HCV-RNA was next tested by means either of 'home-made' nested RT-PCR procedure or the RT-PCR automatic Cobas AmplicorTM Hepatitis C assay, ver. 2.0. In case of an inhibition of PCR detected in the first run of automatic assay, both PCR procedures have been repeated. RESULTS: In 5 cases of automatic assay an inhibition of PCR reaction has been detected in the first run, the RT-PCR procedures has been then successfully repeated in the second run. The presence of HCV-RNA in the liver tissues was detected in a total of 22 cases (69%) by mean of automatic assay and in a total of 20 cases (63%) by means of 'home-made' RT-PCR procedure. Except for two cases of HCV-RNA positivity in a biopsy tissue, detected by means of automatic but not 'home-made' procedure, the results obtained employing the methods have been identical. In one serum HCV-RNA positive case, the presence of HCV RNA has not been detected in a liver tissue using both methods. CONCLUSIONS: Our procedure of RNA isolation combined either with automatic Cobas AmplicorTM Hepatitis C assay, ver. 2.0 or 'home-made' RT-PCR procedure may be useful for establishing presence of HCV-RNA in liver tissue. In this combination, the automatic assay seems to be more sensitive than the 'home-made' procedure.

Biopsy↗

PubMed2001

Acute haemolytic syndrome and liver failure as the first manifestations of Wilson's disease.

Acute liver failure and haemolytic syndrome appeared quite suddenly as the first manifestations of Wilson disease (WD) in five of our patients previously regarded as healthy persons (although an interview showed that 2-4 weeks prior to the illness the patients complained of several non-specific symptoms, such as abdominal pain, headaches, fever, weakness or behavioural changes). All the patients were young women (17-23 years), none of them had any history of liver disease. They were admitted with icterus, nausea, vomiting and symptoms of increasing haemolysis. The diagnosis of WD was given as disturbed copper metabolism. After a short period of observation ascites and anasarca occurred, haemorrhagic diathesis and other symptoms of liver failure increased. Levels of clotting factors decreased rapidly. Despite treatment with D-penicillamine, plasmapheresis, and symptomatic drugs, three of the women died in irreversible liver coma, due to the unavailability of liver transplantation. The fourth woman was carried to the Transplantation Centre, due to aggravation of the symptoms of liver failure, where liver transplantation was performed. Histopathologically micronodular cirrhosis was shown in all these cases. The fifth patient survived having undergone the above treatment without liver transplantation. The main differences between the patient who survived and those who died or underwent transplantation were relatively higher activity of alkaline phosphatase (26 U/l vs. 10-20 U/l), slightly higher levels of clotting factors and prothrombin time, which never fall below 68% of the control (versus 14-44% in other patients). Only in the surviving patient was the Kayser-Fleischer ring present. In four of our patients we found family members who were carriers of WD.

Adolescent↗

PubMed2001

Effect of high fiber vegetable-fruit diet on the activity of liver damage and serum iron level in porphyria cutanea tarda (PCT).

BACKGROUND: During the treatment of coronary heart disease with a vegetable-fruit diet, we have observed the positive effect of the treatment on PCT patients. Therefore, we have now examined the short-term results of the diet on the selected PCT activity parameters. The study was approved by our Review Board. MATERIAL AND METHODS: A group of 13 male PCT patients (mean age 52 years) was evaluated. We assessed the body mass index (BMI), serum iron level, activity of transaminases (ALT, AST), severity of skin symptoms, and urinary prophyrins excretions, before and after a three-week period of vegetable-fruit diet. The diet was of natural vegetable/fruit products, and its daily caloric content was ca. 500 kcal/day. RESULTS: The mean BMI before and after the diet period were 26.8 +/- 4.7 vs. 25.8 +/- 4.3 (p = 0.001), the serum activities of ALT 122.0 +/- 60.7 U/l vs. 75.6 +/- 31.8 U/l, and of AST 91.8 +/- 56.0 U/l vs. 55.2 +/- 14.2 U/l (p = 0.001), respectively. The mean serum iron levels were 188.6 +/- 75.7 mg/dl vs. 140.2 +/- 56.4 mg/dl, serum ferritin concentrations 574 +/- 351 vs. 499 +/- 340 ng/ml (p = 0.04), respectively. Severity of skin lesions and urinary coproporphyrins excretion were significantly diminished during the diet; urinary uroporphyrins excretion was also lowered, but not to a statistically significant level. CONCLUSION: In our group of PCT patients, we noticed the beneficial effect of the vegetable-fruit diet on selected disease parameters. The diet may be useful in the treatment of PCT and diseases associated with PCT.

Adult↗

PubMed2000