Reduced gene expression of bikunin as a prognostic marker for renal cell carcinoma.

AIM: Experimental and clinical studies showed that bikunin, a Kunitz-type protease inhibitor, found in urine and amniotic fluid has a role in spread of tumor cells by providing a significant reduction in the levels of urokinase-type plasminogen activator (uPA) and its specific receptor urokinase-type plasminogen activator receptor (uPAR). The aim of this study was to investigate expression of bikunin at the mRNA level and screen for mutations in exon sequence in renal cell carcinoma (RCC) tissues. MATERIALS AND METHODS: Total RNA and DNA were extracted from paired normal and tumor tissues of total 50 RCC (11 papillary, 8 chromophobe, 26 clear cell, and 5 other types) patients (23 females, mean age: 53.55 ± 14.17; 27 males mean age: 62.1 ± 7.92). Bikunin mRNA levels were detected using semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Mutational screening was performed by using single strand conformation polymorphism (SSCP) method and nucleotide sequence analysis. RESULTS: There was a statistically significant decrease in the 25 (50%) of tumor tissues comparing to normal tissues in terms of mRNA levels of bikunin (Wilcoxon signed rank test, p = 0.0337). According to the classification based on subtypes of RCC; clear cell RCC samples displayed a reduced gene expression (p = 0.0148). Additionally, the patients with the age above 50 had low bikunin expression. The SNP rs80057939 spanning 4(th) exon of bikunin was detected in 13 tumor tissues. However, it was not statistically significant (p > 0.05). CONCLUSION: Decreased bikunin mRNA level in renal cells might be associated with poor prognosis of renal carcinoma. Therefore, gene constructs or exogenous administration of bikunin might be a potential adjuvant therapy for RCC treatment.

Occupational lead exposure effect on liver functions and biochemical parameters.

The effect of occupational lead exposure on the liver function and on the blood biochemical parameters among the battery workers and the muffler repair workers was studied. The study included 22 battery and 38 muffler repair workers. Whole blood lead levels were determined by atomic absorption spectrophotometers. Total protein, albumin, globulin, cholesterol, triglyceride, total bilirubin, aspartate transaminase (AST), alanine transaminase (ALT), gamma glutamyltransferase (GGT), lactate dehydrogenase (LDH), and alkaline phosphatase (ALP) levels were determined in the serum by spectrophotometry. The blood lead levels of the battery workers, muffler repair workers, and the controls were found to be 36.83 +/- 8.13 microg/dL, 26.99 +/- 9.42 microg/dL, and 14.81 +/- 3.01 microg/dL, respectively. Blood lead levels of the workers were significantly higher than those of controls (p < 0.001). The lead level of the battery workers was also significantly higher than that of muffler repair workers (p < 0.001). Although, statisticly significant, higher blood lead levels are not related to toxicity for battery and muffler repair workers. Total protein, globulin, cholesterol, LDH, and ALP levels were within normal levels, however, they were slightly higher than the control levels. Increased LDH among the workers seems to be related rather to other causes than to the liver injury.

Blood lead levels of the battery and exhaust workers and their pulmonary function tests.

In an attempt to understand the impact of inhaled lead on the pulmonary functions, we assessed the blood lead levels and pulmonary functions of the battery and exhaust workers who are potential candidates for lead inhalation. The hospital staff served as control group. The measurements of lead levels were performed using atomic absorption spectrophotometry. Pulmonary function tests (PFTs) were performed using a portable spirometer. The blood lead levels of the battery workers, exhaust workers and controls were found to be 36.83, 26.94 and 14.81 microg/dl, respectively. The values of the workers were significantly higher than the controls (p < 0.001). The lead levels of the battery workers were also significantly higher than the exhaust workers (p < 0.001). PFT results (maximum voluntary ventilation, forced expiration flow and first forced expiration volume values of the workers) were significantly worse than the controls, and their pulmonary function test results were consistent with restrictive problems in the airways.

The relationship between expression of p53/bcl-2 and clinicopathological criteria in endometrioid adenocarcinomas of the endometrium.

OBJECTIVE: Thirty patients with endometrioid adenocarcinomas of the endometrium who had undergone total abdominal hysterectomy and bilateral salpingo-oophorectomy were reviewed histopathologically. MATERIALS AND METHODS: Tissues were stained immunohistochemically with p53 and bcl-2, respectively. Ten cases were selected as grade 1, ten cases grade 2 and ten cases grade 3. RESULTS: We observed 14 cases (46.6%) of bcl-2 and four cases (13.3%) of p53 positivity; p53 expression showed an opposite correlation to bcl-2 staining, estrogen and progesterone. CONCLUSION: We observed a decrease in bcl-2 reactivity and an increase in p53 positive staining as the histological differentiation worsened.

The relationship between expression of p53/bcl-2 and clinicopathological criteria in cervix squamous cell carcinomas.

OBJECTIVE: Thirty cases with squamous carcinomas of cervix who underwent total abdominal hysterectomy and bilateral salphingo-oopherectomy were reviewed histopathologically. MATERIALS AND METHODS: The tissues were stained immunohistologically with p53 and bcl-2. Ten cases were selected as well differentiated (large cell keratinizing) carcinoma, ten cases moderately differentiated (large cell non keratinizing) carcinoma and ten cases poorly differentiated carcinoma (includes small cell squamous cell carcinoma). RESULTS: We observed 11 cases (36.6%) with bcl-2 and 13 cases (43.3%) with p53 positivity. P53 expression showed the opposite correlation to bcl-2 staining. CONCLUSION: A decrease in bcl-2 reactivity and an increase in p53 positive staining was noted as the histological differentiation worsened.

Formalin fixing and paraffin embedding may lead to extra band development in PCR.

The molecular biological analysis of infectious agents requires the availability of a reliable source of microorganisms to be used to recover DNA. Clinical samples can be obtained directly from infected patients or can be propagated using in vitro or in vivo systems. However, repeated sampling from patients is not always possible as the procedure may be invasive or unpleasant, or it is not possible to catch the same agent at the time of second sampling. Moreover, the techniques used may also produce false-positive and false-negative results. We therefore studied the impact of formalin-fixing and paraffin embedding on tissue sampling, and the methodologies such as DNA isolation and PCR amplification of DNAs from archival materials in the diagnosis of Mycobacterium tuberculosis. PCR analyses were done according to standard methods with some modifications. Demonstration of mycobacteria was successful both in tissue sections of the formalin-fixed lymph nodes and in stained fresh materials from patients. However, the results showed the presence of two extra bands in the gel. We accounted for extra band development due to the harshness of the methodology used to isolate nucleic acids from formalin-fixed and paraffin embedded tissue samples or the nature of the fixation procedure, or because of the time passed during storage in which alteration in the chromosomal DNA would take place. Thus, if disease- and tissue specific morphological features, such as sample size, type of fixation, and intralesional heterogeneity are ignored, errors because of sampling and methodologies used may lead to false-positive and false-negative results.

Bernard-Soulier syndrome in a Turkish family.

This report describes the first Turkish family to be diagnosed with Bernard-Soulier syndrome. The family consists of nine members (two parents, three sons and four daughters). The parents were first cousins. The index case, a 22 year-old-man, had a history of haemorrhagic diathesis with thrombocytopenia, giant platelets in the peripheral blood smear and a prolonged bleeding time. Refractory idiopathic thrombocytopenic purpura had been diagnosed elsewhere and a splenectomy had been performed six months previously. Ristocetin agglutination of platelets was defective and flow cytometry analysis of platelet membrane glycoprotein showed markedly reduced expression of glycoprotein lb (2.1%). Bernard-Soulier syndrome was diagnosed. Increased mean platelet volume was found in both parents, one son and three daughters. The other son and daughter were normal.

Dermatoglyphic analyses in children with cerebral palsy.

This study was intended to elucidate the diagnostic values of dermatoglyphic features on the 45 cerebral palsy (CP) patients (28 boys and 17 girls). There were 50 healthy children in the control group. Dermatoglyphic samples were obtained from the both groups by using the paper and ink method and than analysed. The types of dermal patterns of fingertips, the counts of total ridges, the counts of a-b ridges, the values of atd angles, presence or absence of dermal patterns in the hypothenar, thenar/I, II, III, IV interdigital areas, presence of absence of the palmar flexion lines, were compared between the children with CP and control group. It was found that arch, radial loop, whorl prints have increased and ulnar print has decreased in boys investigated which was significant statistically (p < 0.001). No difference was found between investigation and control groups of girls (p > 0.05). The total ridge counts in boys and girls of the investigation group were found significantly decreasing according to the control group (p < 0.001). There was an important decrease in the counts of a-b ridges of investigation group as compared to controls. It was significant in boys (p < 0.01) but not in girls (p > 0.05). The values of atd angles of the investigation group have increased in the control group (p < 0.001 in girls and p < 0.01 in boys). The dermal prints in the hypothenar, thenar/I, II, III and IV interdigital areas showed important differences in the investigation group when compared with the control group (p < 0.01). No clear distinction occurred between the two groups from the viewpoint of palmar flexion lines (p > 0.05). In conclusion, remarkable differences in comparison to controls were found in the dermatoglyphic features of CP cases. In our opinion, by undertaking more studies on the subject and examining a higher number of cases it will be possible to obtain useful data in CP cases indicative of etiologically.

Evaluation of non-invasive clinical samples in chronic chlamydial prostatitis by using in situ hybridization.

Seventy-eight non-invasive prostate specimens collected from patients with chronic non-bacterial prostatitis were evaluated by in situ hybridization (IH) for evidence of Chlamydia trachomatis. Intracellular Chlamydia bodies were detected in 18 of 78 cases (20.6%). Homogeneous blue-black bodies in the cellular cytoplasm were accepted as in situ positive. Chlamydial antigen detected by enzyme immuno assay (EIA) was positive in 12 cases (13.7%), but only nine of them were positive by IH. Our study confirms previous reports implicating C. trachomatis as an aetiological agent in chronic non-bacterial prostatitis, and underscores the applicability of DNA probes for detection and identification of C. trachomatis in prostatic materials.

Effects of ciprofloxacin on chromosomes, and hepatic and renal functions in rats.

In the present study, ciprofloxacin, a fluoroquinolone antibiotic, was evaluated in vivo. The toxic effects of ciprofloxacin (20 and 200 mg/kg/day) on kidney and liver functions, and on chromosomes were investigated in rats. 20 mg/kg/day ciprofloxacin did not significantly affect urine composition, serum enzyme levels and the haematocrit, affecting only creatinine clearance. The 200 mg/kg/day dose increased urine volume, serum creatinine and and creatinine clearance. Ciprofloxacin at both doses had no significant numerical effect on chromosomes, but there was a statistically significant dose-dependent increase in chromatid-type breakage. Significant histological changes in the liver and kidney were not observed.