Ovarian small cell carcinoma of hypercalcemic type - evidence of germline origin and SMARCA4 gene inactivation. a pilot study.

Ovarian tumors from two patients, compatible by histological and immunohistochemical criteria with small cell carcinoma of hypercalcemic type (SCCHT) (WT1+, EMA dispersed+, synaptophysin+ or dispersed+), were extensively sampled in order to find clues to their histogenesis. Subsequently, small foci of immature teratoma were found in both of them (in 1/122 and in 3/80 tumor sections). In one case, microfoci of yolk sac tumor were also present within the teratoma area as well as in the background of the small cell tumor population - in the primary tumor and in omental metastasis. We found a resemblance of the microscopic patterns of SCCHT and atypical teratoid/rhabdoid tumor (AT/RT) of the central nervous system, and this prompted us to evaluate INI-1 and SMARCA4 immunohistochemical expression, because their alternative loss is regarded as a molecular hallmark of AT/RT. INI-1 expression was retained, while that of SMARCA4 was lost. We therefore analyzed tumor DNA by PCR amplification and sequencing for mutations in the SMARCA4 gene (NG_011556.1), which were identified in both tumors (c.2184_2206del; nonsense c.3277C>T - both in one tumor; nonsense c.3760G>T in another tumor). These data suggest that SCCHT is most likely an embryonal tumor originating from immature teratoma and related to malignant rhabdoid tumor. Further analyses are necessary to determine whether the tumors diagnosed as SCCHT constitute a homogeneous group or represent more than one entity.

TP53 status determines clinical significance of ERBB2 expression in ovarian cancer.

ERBB2 expression has been found in 19 to 44% of ovarian carcinomas; however, its predictive value has not been demonstrated, and trastuzumab has not found clinical application in ovarian cancer patients. We evaluated clinical significance of ERBB2 expression in relation to TP53 accumulation in ovarian carcinoma patients treated with platinum-based regimens. Immunohistochemical analysis with CB11 and a novel NCL-CBE356 antibody (against the internal and external domains of ERBB2, respectively) was performed on 233 tumours (FIGO stage IIB-IV); the US Food and Drug Administration-approved grading system with 0 to 3+ scale was used for evaluation, and the results were analysed by the Cox and logistic regression models. In all, 42% of the tumours expressed (category 1+, 2+ or 3+) either CB11 or CBE356 or both (CB11/CBE356 parameter). Associations between ERBB2 expression and clinical factors were observed only if tumours with staining category 1+ were grouped together with tumours showing staining categories 2+ and 3+. CB11/CBE356 parameter had a better predictive value than CB11 alone. CB11/CBE356 expression was negatively associated with platinum sensitivity (PS) in the TP53(-) group (P=0.022) and with disease-free survival (DFS) in the TP53(+) group (P=0.009). Our results may suggest that trastuzumab should be given postoperatively to patients with TP53(-)/ERBB2(+) ovarian carcinomas to enhance PS, and after completion of chemotherapy to patients with complete remission and TP53(+)/ERBB2(+) carcinomas to extend DFS time (in total to 30.4% of all patients analysed). Thus, novel criteria for ovarian cancer patient inclusion for clinical trials with trastuzumab should be considered and tested.

Skin reactivity to autologous bacteria isolated from respiratory tract of patients with obstructive pulmonary disease.

Bacterial flora of various strains was isolated from sputum and in some cases from BAL fluid of 75 patients with obstructive pulmonary disease experiencing dyspnea symptoms accompanying infections of respiratory tract. Among strains recognized traditionally as pathogenic in respiratory tract, we also isolated various strains typically called "normal oropharyngeal flora", "physiologic" or "non-pathogenic" bacteria. Those latter strains used in the skin tests in autologous manner for each patient had the property of provoking early (15 min) and late (24-48h) reactions. Early reactivity resembles that induced in the tests with airborn allergens. This suggests a potentially important role of those currently ignored strains in pathogenesis of obstructive pulmonary disease.

P21WAF1, P27KIP1, TP53 and C-MYC analysis in 204 ovarian carcinomas treated with platinum-based regimens.

BACKGROUND: The prognostic and predictive value of cell cycle regulatory proteins in ovarian cancer has not been established. We evaluated the clinical and biological significance of P21(WAF1), P27(KIP1), C-MYC, TP53 and Ki67 expressions in ovarian cancer patients. MATERIALS AND METHODS: Immunohistochemical analysis was performed on 204 ovarian carcinomas of International Federation of Gynecology and Obstetrics (FIGO) stage IIB to IV treated with platinum-based chemotherapy. Multivariate analysis with Cox and logistic regression models was performed in the whole group, and in the TP53-negative and TP53-positive subgroups. RESULTS: High P21(WAF1) labeling index (LI) was an independent positive predictor of platinum-sensitive response (P = 0.02). Overall survival was positively influenced by P21(WAF1) LI (P = 0.02) or by P21(WAF1) plus P27(KIP1) LI (P = 0.004) in the TP53-negative group only. Ki67 LI showed borderline association with disease-free survival (P = 0.05). Growth fraction was negatively associated with P21(WAF1) and P27(KIP1) indices in the TP53-negative group (P = 0.023 and 0.008, respectively), and these associations were borderline or lost in the TP53-positive group. Endometrioid and clear cell carcinomas differed from other carcinomas by having a low incidence of TP53 accumulation, a high incidence of C-MYC overexpression (70%) and a low median Ki67 LI (all with P <0.001). CONCLUSIONS: We have shown an independent predictive value of P21(WAF1) LI in ovarian carcinoma patients. The prognostic value of P21(WAF1) and P21(WAF1) plus P27(KIP1) LI was determined by TP53 status. A high frequency of C-MYC overexpression in endometrioid and clear cell carcinomas may suggest its role in the development of these tumor types.

Nijmegen breakage syndrome gene (NBS1) alterations and its protein (nibrin) expression in human ovarian tumours.

We looked for NBS1 gene (602667) alterations and changes in nibrin expression in 162 human gynaecological tumours, mostly ovarian. Exons 6-8 and 10 of the NBS1 gene were evaluated by the SSCP and direct sequencing method. Nibrin expression was detected immunohistochemically with the use of the p95NBS1 (Ab-1) antibody. The 657del5 mutation (Slavic mutation) was found in two of 117 carcinomas studied (1.7%) - in both cases it was present in the germline; one of these tumours showed loss of heterozygosity (LOH) for the 657del5 mutation and loss of nibrin expression. We have found three types of novel germline intron variants: (1) two concomitant transitions (G to A) at bases 14009 and 14256; (2) C to T transition at base 13998; (3) G to C transversion at base 20035. Among the carcinomas studied, the intron variants were associated with a clear cell histological type (p = 0.004). Our results may suggest that NBS1 gene alterations contribute to the development of rare ovarian carcinomas. LOH for 657del5 in tumour tissue may support the hypothesis that the NBS1 gene functions as a tumour suppressor.

[Analysis of HLA class I and II antigens composition in high-risk pregnancy]. / Analiza wystepowania antygenów HLA klasy I i II w ciazy wysokiego ryzyka.

DESIGN: We have analyzed the frequency of HLA class I and II antigens in high-risk pregnancy. MATERIAL AND METHODS: Altogether, 22 gravida hospitalized at the Department of Pathology of Pregnancy and Labour, Pomeranian Medical University in Szczecin formed group I (PE) with 12 cases of preeclampsia and group II with 10 cases of the antiphospholipid syndrome (APS). The control group included 40 multigravida. Typing of HLA class I and II antigens was done using the two-stage microcytotoxic test (NIH) of Mittal. RESULTS: Antigen B35 was found more frequent in preeclampsia and antiphospholipid syndrome groups than in multigravida. Furthermore, a statistically significant difference in the frequency of homozygotes for the HLA-DR locus was noted between groups PE and APS on one hand, and controls on the other. CONCLUSIONS: Identical HLA-DR3, DR4 and DR5 antigens were found more frequently in preeclampsia, while identical DR4 and DR6 in the antiphospholipid syndrome.

[The role of bacteria in allergic and pseudoallergic reactions]. / Rola bakterii w reakcjach alergicznych i pseudoalergicznych.

The variety of microbial pathogenic factors and relations between human and bacterial cells in allergo-inflammatory diseases are presented. A special attention have been paid to sugar-lectin interactions in pseudoallergic processes. The role of specific antibodies as modifiers of basophils releaseability is considered.

Polymorphism of the fourth component of complement (C4) and three clinical pictures of hepatitis B virus infection in children.

The fourth component of complement (C4) phenotype and allele frequencies were studied in 3 groups of unrelated children affected with hepatitis B virus (HBV) infection (25 with glomerulonephritis (GN), 39 with chronic active hepatitis (CAH), 24 with Gianotti-Crosti syndrome (GS-S)) and in 100 unrelated healthy individuals (H). Ten phenotypes in GN, 14 in CAH, 10 in GC-S and 23 in H were found. Some rare phenotypes appeared in the individual cases of ill children. Statistical analysis was performed by chi-square test. Each group was compared to the others. We found significantly higher frequency of the A3B2,Q0 phenotype in CAH than in H as well as significant increase of the A x 4 gene frequency in GC-S as compared to H. However, these events could be caused by limited number of tested patients. We suspect that the association between C4 alleles and immune-complex diseases, studied here, does not rather exist.

Frequency of complement third component (C3) and properdin factor (BF) phenotypes in patients with various clinical manifestations of HBV infection.

Four groups of children were tested for the distribution of C3 and BF phenotypes: HBV-infected patients with Gianotti-Crosti syndrome, with CAH and with glomerulonephritis, and healthy children. The frequency of the phenotype C3F was significantly higher in children with Gianotti-Crosti syndrome in comparison with healthy children. The frequency of the phenotype BFS was significantly higher in patients with glomerulonephritis than in individuals with CAH. The difference between the frequency of the BFS phenotype in glomerulonephritis patients and that in healthy subjects neared significance. We suggest that the carriers of these phenotypes are characterized by susceptibility to some immune complex diseases associated with HBV infection.

[Bacterial flora of acne lesions in diagnostic material of the Microbiology and Immunology Department, Pomeranian Medical Academy (PAM) in Szczecin]. / Flora bakteryjna zmian tradzikowych w diagnostycznym materiale Zaklad Mikrobiologii i Immunologii PAM w Szczecinie.

A retrospective analysis of the bacterial flora of acne lesions was carried out in 320 patients. Atypical flora was isolated very rarely. The pathogenic flora was found most frequently in the Autovaccine Laboratory when the microbiological material was taken directly onto plates and the investigation was repeated several times. Atypical flora was isolated only from three cases. Propionibacterium and Staphylococcus epidermidis were found to be sensitive to the antibiotics routinely used in the treatment of acne. The highest antibacterial effect in vitro against both these species was demonstrated using rifampicin and tetracycline.

Frequency of Gm and Km phenotypes in children with mumps meningitis.

Children with mumps meningitis, with mumps without meningitis and healthy subjects were examined for Gm 1, 2, 3, 5, 21 allotypes and for the Km 1 allotype. The comparison of the frequencies of Gm and Km phenotypes in three tested groups (each to the other) did not reveal significant differences. An association between mumps meningitis and the Gm or Km phenotypes was not found.

Prognostic value of an assessment of ABH(0) isoantigens and Thomsen-Friedenreich (TF) antigen expression in patients with urinary bladder tumours.

The discriminating and prognostic value of the expression of ABH(0) and TF antigens in tumour cells of 76 patients with urinary bladder tumours was evaluated. In patients with superficial tumours the most frequently defined phenotype was ABH(-) TF(+) and in those with deep tumours ABH(-) TF(-). The lowest frequency of recurrences was seen in patients with TFCr(+) antigenic phenotype and no patient died in this group. The mean lifespan of patients with TF(-) tumours was shorter (21 months) than of those with TF(+) tumours. For detection of tumours with higher risk of recurrences an analysis of the combination of markers revealed to be more predictive than analysis of a single marker.

Gm and Km phenotype frequencies in children with chronic active hepatitis (CAH) B virus infection.

Patients with CAH, extrahepatic HBV manifestation and healthy children were studied for presence of Gm 1, 2, 3, 10, 21 factors and Km 1 factor. significantly higher frequency of Gm (1, 2, 3, 10, 21) phenotype was shown in CAH group as compared with the other two groups. Relationship between Km factors and examined groups was not found.

[Comparison of bacterial flora isolated from the urinary tract of patients examined in the Department of Microbiology and Immunology of the Pomeranian Academy of Medicine and from the bacteriology laboratory of the Municipal Health Care Group in Szczecin]. / Porównanie flory bakteryjnej hodowanej z dróg moczowych pacjentów badanych w Zakladzie Mikrobiologii i Immunologii Pomorskiej Akademii Medycznej i Pracowni Bakteriologicznej Miejskiego Zespolu Opieki Zdrowotnej w Szczecinie.

The authors have analyzed 5305 urine samples from patients with significant bacteriuria, examined in 1988-1992 at the Department of Microbiology and Immunology of the Pomeranian Academy of Medicine, performing analyses for the State Clinical Hospital No. 2 in Szczecin, and in the Bacteriology Laboratory of the Municipal Health Care Group in Szczecin. The results obtained demonstrate a dominance of Escherichia coli among patients of general medicine ambulatories and internal medicine departments (70%), while patients of the urologic department and urologic out-patient clinic were positive for this bacteria in only 30-40%. It should be mentioned that a significant increase in the number of infections by Pseudomonas, Acinetobacter, and Serratia strains has been observed, mainly among patients after urologic operations.

[Biocenosis of the vagina and infection with Chlamydia trachomatis]. / Biocenoza pochwy a zakazenie Chlamydia trachomatis.

The authors have examined 379 women for infection by Chlamydia trachomatis, performing vaginal smears and routine bacterial cultures. C. trachomatis was discovered in 33.2% of the women, most frequently accompanied by erosion of the collum, cervicitis and CIN-Ca. Women with C. trachomatis had the following levels of vaginal purity: 1-44.4%, 2-23.0%, 3 and 4-29.4%, 3.2% -leukocytes only. Leukocytes were present in significantly more women with level 1 of vaginal purity than in the control group. Additional infections (Gardnerella vaginalis, Streptococcus agalactiae, Candida albicans, Escherichia coli and Trichomonas vaginalis) were found in only a small percentage of women with C. trachomatis, just as in controls. The presence of leukocytes or level 1 of vaginal hygiene with leukocytes may favour C. trachomatis, just as in controls. The presence of leukocytes or level 1 of vaginal hygiene with leukocytes may favour C. trachomatis infection, therefore these cases should be subjected to a more thorough examination.

Phenotype distribution of the third component of complement (C3) and of the properdin B factor (BF) in children with mumps meningitis.

The C3 and BF phenotype frequencies were studied in children with mumps meningitis. No significant differences were found between this group and other groups; children with mumps without meningitis and healthy children.

An immunoblotting procedure for screening glycophorins and band 3 protein in the same blots. Identification of glycophorin and band 3 variant forms.

An immunoblotting procedure is described which makes it possible to screen multiple blood samples for the presence of glycophorin and band 3 variant forms with altered electrophoretic mobility. The procedure can be simplified by using whole red blood cell hemolysates instead of membranes for SDS-polyacrylamide gel electrophoresis. The use of hemolysates also has the advantage that antigens sensitive to proteolysis are not degraded in vitro. The same nitrocellulose blots were used for immunoenzymatic detection of glycophorins with a set of anti-glycophorin monoclonal antibodies, and for autoradiographic detection of band 3-derived bands with 125I-labeled anti-band 3 monoclonal antibody. The screening of 157 Caucasian blood samples revealed the presence of a slower-migrating form of band 3 in seven cases and variant glycophorin in one case. The variant glycophorin exhibited the features of hybrid glycophorin of B-A type.

[Occurrence of antituberculous antibodies in patients with lung tuberculosis and other inflammatory states of the respiratory system]. / Wystepowanie przeciwcial przeciwpratkowych u chorych z gruzlica pluc i innymi stanami zapalnymi ukladu oddechowego.

In this study an indirect immunofluorescence method was applied. Determinations regarded the level of anti-H37Rv antibodies in patients with bacteriologically confirmed lung tuberculosis and in a control group consisting of healthy volunteers. Antibody levels were significantly higher in sick patients. We conclude that this method may be applied as a supplementary diagnostic test in doubtful cases and that it can prove useful as screening test for investigation of groups of population especially at risk from tuberculosis. Applied method of indirect immunofluorescence is easy to perform, quick (time of the test takes about 3 hours), available in Poland, and cheaper than ELISA.