Prevalence of Chlamydia trachomatis and herpes simplex virus 2 in cervical carcinoma associated with human papillomavirus detected in paraffin-sectioned samples.

PURPOSE: The aim of the study was to evaluate the frequency of occurrence of HPV and co-infection: Chlamydia (C.) trachomatis and HSV-2 in cervical cancer. MATERIAL AND METHODS: The study group consisted of 570 paraffin-sectioned samples of patients with cervical cancer. In order to identify viral and bacterial DNA in DNA isolated from archival, postoperative material, PCR analysis was performed using starters complementary to various types of HPV, HSV-2 and C. trachomatis. RESULTS: In patients with squamous cell cervical cancer the presence of 33 types of HPV was found in 90% (468/520). HPV 16 infections occurred in 69.4% (325/468), while HPV 18 infections were present in 30.5% (143/468) of cases. In the control group C. trachomatis and HSV-2 were observed in four cases (4/50), which constitute 8.0%. In the tissue sections from patients with squamous cell cervical carcinoma, C. trachomatis was identified in 26% (135/520) and HSV-2 in 28% (145/520). In the group of patients with adenocarcinoma C. trachomatis infections were found in 24% (12/50) and herpes virus was identified in 30% (15/50). Statistically significantly higher frequency of occurrence of HSV-2 and C. trachomatis was observed in paraffin-sectioned samples for patients with invasive cervical cancer compared to the control group, without neoplastic lesions (p < 0.05). No correlation was found between frequency of occurrence of HPV and C. trachomatis and of HPV and HSV-2 detected in paraffin-sectioned samples for cervical carcinoma.

RAS inhibitors decrease apoptosis of acinar cells and increase elimination of pancreatic stellate cells after in the course of experimental chronic pancreatitis induced by dibutyltin dichloride.

Chronic pancreatitis (CP) is a progressive disease, in which the exocrine function of the gland is gradually lost and fibrosis develops due to repeated episodes of acute pancreatitis. The aim of the study was to investigate the effects of RAS inhibitors on the apoptosis of acinar cells and pancreatic stellate cells (PSCs) elimination in experimental CP induced by dibutyltin dichloride (DBTC). CP was induced by administration of DBTC to the femoral vein. Simultaneously captopril, losartan, enalapril and lisinopril were administered intraperitoneally. The rats were decapitated after 60 days and tissue of pancreas was collected. In rats treated by DBTC the features of inflammatory infiltration, ductal lumen dilatation, fibrosis were found. Strong reactivity with caspase2(L) and clusterin-beta antibodies was observed in areas of fibrosis. In animals treated with RAS inhibitors inflammatory changes and fibrosis were less severe. In groups of rats treated with DBTC and RAS inhibitors immunoreactivity of caspase(2L) and clusterin-beta was weak. Positive immunostaining against smooth muscle actine and desmin was observed in the elongated cells (PSC-s). This reaction was weak in groups of rat treated with DBTC and RAS inhibitors. Treatment of CP rats with RAS inhibitors alleviate apoptosis of pancreatic acinar cells and induces PSCs elimination.

Chlamydia trachomatis and herpes simplex virus 2 infection in vulvar intraepithelial neoplasia associated with human papillomavirus.

BACKGROUND: The role of viral and bacterial co-infection is stressed in VIN. A view that VIN is a sexually transmitted disease made the area of research larger and stimulated scientists to seek other sexually transmitted factors, among which Chlamydia trachomatis and Herpes simplex are frequently examined. PURPOSE: The aim of the study was to evaluate the frequency of occurrence of HPV DNA and the frequency of co-infection with Herpes virus type 2 and Chlamydia trachomatis in VIN. MATERIAL AND METHODS: We identified archival diagnostic phase tissue specimens from 41 cases of vulvar intraepithelial neoplasia III. From the same paraffin blocks containing material from the margins of surgical sections during vulvectomy, normal epithelial tissue fragments were collected. They constituted the control group. Lesion characteristics were examined in comparison with the presence of HPV DNA, HSV-2 and Chlamydia trachomatsis. Identification was performed using PCR. RESULTS: In the study group HPV infection was found in 75.6% of cases. In 73% of cases it was HPV 16. In the control group we found HPV 16 DNA in only one case (2.43%). In the HPV positive study group HPV 16 was found in 30 (30/31) cases. In only one case (1/31) it was HPV 18 type. In the study group of 41 cases with VIN, HSV-2 infection was found in six cases (14.63%). In comparison with the control group (9.75%) the difference was not statistically significant. The frequency of occurrence of Chlamydia trachomatis in the analyzed study material was 14.63% (6/41) and in the control group it was 9.75% (4/41). The difference was not statistically significant. Statistical analyses of correlations between the occurrence of DNA HPV and HSV-2 as well as of HPV and Chlamydia trachomatis showed no correlation in either case. CONCLUSION: No correlation was found between the frequency of occurrence of HPV and HSV-2 and HPV and Chlamydia trachomatis in either group.

The role of adenosine A2a receptors in experimental acute pancreatitis.

PURPOSE: The role of adenosine and its receptors in acute pancreatitis remains unelucidated. The aim was to evaluate the effects of the adenosine A2a receptor agonist and antagonist in the severe, taurocholate-induced experimental acute pancreatitis (EAP). MATERIAL AND METHODS: The experiments were performed on 80 male Wistar rats, subdivided into 4 groups: C--the control rats, I--the EAP group, IIA--EAP group treated with the A2a adenosine receptor agonist CGS 21680, IIB--EAP group treated with the A2a adenosine receptor antagonist ZM 241385. The blood for alpha-amylase and lipase and tissues samples for the morphological examinations and immunohistochemistry for A2a receptors were collected in 2, 6, 24 hours of the experiment. RESULTS: The serum alpha-amylase tended to decrease in the group IIA as compared to EAP untreated after 6 and 24 h. No significant effect of both treatments on serum lipase was noted. The administration of CGS 21680 resulted in favorable decrease of the inflammatory cell infiltration, hemorrhagic changes, necrosis and vacuolization of acinar cells, without an evident effect on the edema of the interstitial tissue. The administration of ZM 241385 did not affect the scores of necro-hemorrhagic changes and inflammatory infiltration, whereas it decreased the scores of vacuolization and edema. In all groups the expression of A2a receptors was similar. CONCLUSIONS: Our findings suggest, that A2a adenosine receptors are involved in the course of sodium taurocholate EAP. It is probable that the modulation of some subgroups of adenosine receptors could alleviate the course of severe experimental AP.

K-ras gene point mutations and p21ras immunostaining in human ovarian tumors.

It is well recognized that genetic alterations within oncogenes, tumor suppressor genes, DNA mismatch repair and excision repair genes contribute to tumorigenesis within the human ovary. This study was undertaken to screen for the existence of K-ras gene point mutations in paraffin-embedded slides randomly selected from benign and malignant ovarian tumors applying the PCR-RFLP technique. Expression of p21ras was also assessed in 30 primary ovarian adenocarcinomas immunohistochemically. K-ras codon 12 point mutations occurred in nine of 40 (22.5%) cases. They were not identified in two benign mucinous cystadenomas, but in one out of two (50%) mucinous tumors of LMP (low malignant potential), in five out of 30 (17%) ovarian adenocarcinomas, and in one case of adenocarcinoma metastatic to the ovary. K-ras activation was also detected in one out of four (25%) sex cord-stromal cell tumors (folliculoma), and in one dysgerminoma. None of these tumors exhibited K-ras codon 13 point mutations. Gene alterations were more frequently found in mucinous than in non-mucinous (30% vs 10%) tumors, although the difference did not reach significance (p > 0.05). The frequency of K-ras point mutations was correlated neither with clinical nor with pathological variables of cancer. Cytoplasmic p21ras was expressed in all adenocarcinomas negative for K-ras point mutations, whereas one of five (20%) K-ras-positive tumors exhibited lack of immunoreactivity. In conclusion, these findings confirm the role of K-ras activation in mucinous ovarian tumors. p21ras expression is not necessarily associated with K-ras gene alterations in human ovarian adenocarcinomas.

Content of folic acid and free homocysteine in blood serum of human papillomavirus-infected women with cervical dysplasia.

The authors estimated the concentrations of folic acid and free homocysteine in the blood serum of women with CIN III (cervical intraepithelial neoplasia-Burghard's classification) infected with DNA HPV (human papillomaviruses) of type 16 and/or 18. The control group consisted of 49 patients with normal cytological smears without HPV infection. Types 16 and/or 18 DNA HPV were found in 50 patients. This women qualified for the studied group. The sequence of DNA HPV type 16 and/or 18 was identified with the PCR method (polymerase chain reaction). The high-performance liquid chromatography (HPLC) method was employed to evaluate the levels of folic acid and free homocysteine in the blood serum of the examined patients. Significantly lower levels of folic acid and higher levels of free homocysteine were observed in the blood serum of HPV-positive patients with CIN III. The correlation was found between serum concentrations of folic acid and free homocysteine in both groups.

Principles of induction chemotherapy for non-small cell lung cancer.

The results of lung cancer treatment have not significantly improved for many years. About 35% of patients with non-small cell lung cancer (NSCLC) are in clinical stage IIIA. Clinically asymptomatic distant metastases occur in the majority of these patients. In such cases only combined treatment offers a chance of cure. In the Chest Surgery Center in Lublin a clinical trial was carried out aimed to assess late results of combined treatment in patients with IIIA NSCLC. Over 700 patients were enrolled in the study. The results of the trial disclosed, that neoadjuvant chemotherapy prolonged life of the operated patients and improved their life quality. However, a question of qualification for this complex treatment and complexity of assessment criteria, still remain to be answered.

Clinical and morphological analysis of 143 cases of "pulmonary peripheral shades" in patients treated in the department of thoracic surgery of Medical Academy in Lublin.

In spite of a constant development of various diagnostic methods, in many cases of "peripheral shades" the exploratory opening of the chest appears to be the only way leading to a correct diagnosis. In this study, 143 patients were examined. These patients were referred to the Thoracic Surgery Department because of peripheral shades in the lung areas observed, often accidentally; on regular checkup X rays. Our paper assessed the usefulness of endoscopic examinations in diagnosing the changes situated peripherally in the lungs. 83.2% of those shades were recognized as neoplasms on the basis of histopathological examinations. Moreover , we evaluated the concordance of histopathological diagnosis before and after the operation.

Multifocal fibromatosis: a case report.

We present of rare case of multifocal fibromatosis in a 52 year-old women. In 1996, she was first evaluated for a tumour of the right breast and on the basis of the surgical specimen the extra-abdominal fibromatosis was diagnosed. Four years later, she was reevaluated for the tumor of the right lung, and then in 2001 for the lesion of the right parietal pleura. Microscopic examination of pulmonary and pleural lesions revealed histological pattern almost identical with the breast tumor. The recurrent lesions were located proximally to the primary one.

Sclerosing stromal tumour--a rare ovarian benign neoplasm: description of three cases.

Sclerosing stromal tumour is a rare benign usually unilateral ovarian tumour. We present three cases of this neoplasm in young females, including one bilateral case. All tumours were composed of cellular areas forming pseudolobular patterns, as well as vascular, fibrous, and myxoid areas. The characteristic feature of tumours was cellular heterogenity. All tumours revealed negative immunostaining for cytokeratin, and intense positive immunostaining for vimentin, and smooth muscle actin. Immunostaining for desmin and progesterone receptors was weakly positive.

The coexistence of cystic nephroma and renal cortical adenoma.

Cystic nephroma is rare, but its coexistence with renal cortical adenoma is unique and has never been reported previously. We present a unique case of simultaneous occurrence of cystic nephroma and renal cortical adenoma in 47-year-old man. Both tumours were situated in the lower pole of the right kidney and were clinically asymptomatic.

Expression of p21 and bcl-2 proteins and p53 mRNA in surgically resected preparations of non-small cell lung cancer (stage IIIA) after etoposide and cisplatin chemotherapy.

Apoptosis which is also a called programmed cell death plays an important role during development, homeostasis and in many diseases such as cancer. Apoptosis is a genetically encoded cell death program defined by characteristic morphological and biochemical features. It is well recognized as a distinct pathologic mechanism in tumours responding to anticancer therapies. Many genes play an important role in this process. We evaluated an expression of the tumour supressor gene p53 and proteins p21 and bcl-2 in non-small cell lung cancer. We examined resected tumour tissues from 30 patients who received neoadjuvant chemotherapy. As a control we assessed tissues from patients treated without chemotherapy. Histological slides of the resected tumours were evaluated by TUNEL, in situ hybridisation and with immunoperoxidase staining procedure. The results were documented by photography. We examined the level of extinction using cytophotometry. In conclusion, preoperative chemotherapy induces apoptosis in cancer cells. The level of p53 correlates with the acceleration of TUNEL reaction. The loss of bcl-2 expression correlated with an increased apoptotic cell death. There was an increased p21 protein expression in the examined cancer tissues after chemotherapy.

[Hydrophobic DNA adducts in relationship to estrogen and progesterone receptors content in human uterine cancer]. / Hydrofobowe addukty DNA w relacji do gestosci receptorów estrogenowych i progesteronowego w gruczolakorakach blony sluzowej macicy u kobiet.

OBJECTIVE: Determination of the relationship between hydrophobic DNA adducts (A) and estrogen receptors (ER) and progesterone (PR) receptor status in uterine cancers. METHODS: Using the P1 enriched version of 32P-postlabeling for hydrophobic DNA adducts detection on polyethyleneimine (PEI) cellulose thin layer chromatograms (TLC) we examined 11 uterine cancer DNAs. The quantification of the adducts was performed by Cerenkov counting of the spots. ER and PR status was recognized histochemically and H-score estimate was performed for each investigated cancer tissue. Patterns of uterine cancer DNA adducts were compared to the maps of adducts recognized in normal human endometrium. RESULTS: In three of the studied uterine cancers there was no positive staining of ER and PR; in one case there was a weak ER staining but PR staining was negative. In ER negative tumors the A level was significantly higher than in ER positive cancers (138.1 +/- 64.1 vs. 49.7 +/- 26.8 adducts per 10(9) nucleotides, respectively, p < 0.05). Highest A levels were found in two ER and PR negative G3 metastatic tumors. Finally, in all investigated cancers there was a strong, inverse correlation between ER content and A level (r = -0.67, p < 0.03). In addition, the correlation between PR level and A was of borderline significance (r = -0.6, p = 0.053). The TLC patterns of adducts in uterine tumors were found to be qualitatively similar, but not quantitatively, to those observed in normal human endometrium DNA. CONCLUSION: The data presented suggest that the hydrophobic DNA adducts could play a role in a sex-steroid hormone independence of human endometrial cancers. The highest accumulation of DNA adducts was recognized in neoplasms displaying the most malignant phenotype.

[Overall genomic DNA methylation in relation to estrogen]. / Metylacja genomowego DNA w relacji do gestosci receptora estrogenowego i progesteronowego w gruczolakorakach blony sluzowej macicy u kobiet.

Overall genomic DNA methylation was analyzed using enzymatic digestion into nucleotides, 32P postlabeling, two-dimensional thin-layer chromatography on cellulose plates and phosphobioimaging quantitation, in relation to immunohistochemically measured estrogen (ER) and progesterone receptor (PR) status of 15 uterine cancers. Mean 5-methyldeoxycytosine (m5dC) content did not differ between ER-positive and ER-negative neoplasms. Highest values of m5dC were noted both in ER-negative and ER-positive tumors. Additionally, there was no low DNA methylation in ER negative uterine cancer tissues. Decrease of the overall genomic DNA methylation could be related to the increase of ER/PR ratio, however it was not significant in our investigation. The potential role of steroid receptors status in uterine cancer tissue is discussed.

Immunohistochemical study of p53 in non-small cell lung cancer before and after preoperative chemotherapy.

The purpose of the study was to assess the expression of p53 in non-small cell lung cancer (NSCLC) before and after treatment with cisplatin and vepeside (PE) and to define a relationship between p53 expression and responsiveness to chemotherapy prior to surgery. Material for study consisted of specimens obtained from neoplastic infiltrate before chemotherapy (biopsy material) and tumour specimens obtained after chemotherapy (surgical material). The study population was a group of 35 patients with stage IIIA NSCLC. p53 protein accumulation was detected by immunohistochemistry using antibodies against p53: NCL-p53 (clone BP-53-12) (Novocastra) on paraffin embedded specimens. p53 expression was found in 21 patients (60%) before and after chemotherapy. In 14 patients (40%) p53 negativity was seen both in biopsy and surgical material. The level of p53 staining after chemotherapy as compared with that before treatment changed from -53 to +34. There was a mean increase by 1.52, which appeared statistically accidental (p > 0.70). There was no significant relationship between p53 expression and responsiveness to chemotherapy (from p > 0.33 to p > 0.70) and between the magnitude of changes in p53 expression and response to chemotherapy (p > 0.39). There was also a very low correlation (r to 0.10; p > 0.50) between responsiveness to therapy and p53 negativity.

Expression of PCNA in non-small cell lung cancer before and after treatment with cisplatin and vepeside.

The purpose of the study was to assess expression of the proliferating cell nuclear antigen (PCNA) in non-small cell lung cancer (NSCLC) after administration of cisplatin and vepeside in patients with clinical stage IIIA and to define a relationship between PCNA expression and tumour responsiveness to preoperative chemotherapy. An immunohistochemical study with a mouse monoclonal antibody against PCNA (Novocastra, Clone PC10, IgG2a Class) was performed on paraffin embedded specimens using the ABComplex/HRP method. Material for study was available from 35 patients and consisted of biopsy specimens obtained from neoplastic infiltrate before chemotherapy and tumour specimens obtained from the same patients during surgery 3-4 weeks after chemotherapy. PCNA immunoreactivity was observed in all the cases (100%) both before and after chemotherapy. Despite treatment with cisplatin and vepeside prior to surgery the PCNA index (IPCNA) was significantly higher (p < 0.002) irrespective of tumour responsiveness to chemotherapy. There was a positive correlation (p < 0.04) between tumour size and IPCNA after chemotherapy in a group of patients with a similar extent of neoplastic infiltrate. No correlation was seen between PCNA expression in biopsy specimens and tumour responsiveness to chemotherapy.

Flow cytometric analysis in diagnosis of thymoma metastases.

Thymomas are relatively rare tumours of the anterior mediastinum, constituting approximately 10-15% of all mediastinal tumours. In contrast to other neoplasms, they rarely present distant metastases. We describe a case of thymoma with long survival and skin metastases diagnosed by two-colour flow cytometry.

[Biomechanical investigation of distraction forces appearing in the course of femoral lengthening in rabbits: radiological and histological study]. / Biomechaniczne badania sil dystrakcyjnych wystepujacych podczas wydluzania piszczeli u królików--obserwacje radiologiczne i histologiczne.

This study was conducted in order to determine the values of forces necessary to stretch bony fragments during tibia lengthening in rabbits. The aim of this study was to determine the conditions which lead to complications of the lengthening process related to the rate of distraction. The experiment was conducted on 16 rabbits of New Zealand breed. The construction of the apparatus eliminated tension resulting from the so-called "bending moment". In all rabbits distraction was began on the 5th day after fixing the apparatus. The standard rate of lengthening was 2 x 0.25 mm per day. In 8 rabbits the rate of distraction was regulated. The distraction forces were measured and electronically recorded during the distraction procedure, during the animal's movement and resting position at strictly monitored intervals. Radiological examination of the lengthened tibia was performed once a week. Histological examination was performed in three rabbits after completion of the experiment. We made 328 measurements of the involved distractive forces, collecting a total of 223 MB of data. Graphical representation of the lengthening force and it's performance turned out to be similar for various experimental animals and it's course was typical for every stage of the lengthening process. At the time of distraction of the apparatus the measured force increased on average by 0.87 N (minimum 0.61, maximum 1.1N). Abnormal rates of distraction resulted in a break of the callus continuity in 3 rabbits. Radiological and histological examinations were used to assess to confirm certain observations concerning the technique and rate of distraction.

The diagnostic value of cytomorphological traits in low and high risk type HPV infections.

The purpose of the study was to establish the diagnostic value of cytomorphological traits present in cervical smears in determining HPV infection. We examined 528 women, most of them between 18 and 34 years of age and sexually active, for the prevalence of HPV infection and compared the results of HPV DNA detection with the presence of cytomorphological traits of HPV infection in their cervical smears. The Digene Hybride Capture System technique was employed to detect HPV DNA in cytological material; cervical smears were examined by an experienced pathologist, who did not have prior knowledge of the HPV DNA test results. None of the cytomorphological traits examined proved to be pathognomonic for HPV infection. The diagnosis of HPV infection cannot be determined with certainty by a routine cytological examination. Only 15% of cervical smears from women infected with HPV showed classic cytomorphological traits of HPV infection - koilocytosis and/or dyskeratosis. We conclude that if more than three non-classic traits of HPV infection are found in a cervical smear, HPV infection can be suspected.