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.

Another rare case of a child with de novo terminal 9p deletion and co-existing interstitial 9p duplication: clinical findings and molecular cytogenetic study by array-CGH.

Trisomy 9p is the fourth most common chromosome abnormality found in liveborns. We report on a rare case of partial trisomy 9p complicated by partial monosomy 9p. Clinical manifestation included craniofacial abnormalities typical for trisomy 9p syndrome, developmental delay, mental retardation and brain anomaly in the form of Dandy-Walker malformation. The cytogenetic abnormality was investigated with FISH and array-CGH to characterize the breakpoints of the complex rearrangement. The patient's karyotype was 46,XX,der(9)del(9)(p24)dup(9)(p21p24)dn.arr 9p24.3p24.2 (1-2,414,485)×1,9p24.2p21.3(2,414,485-24,101,280)×3. The cytogenetic rearrangement led to a 2.4-Mb deletion of 9p24.2pter and a 21.6-Mb duplication of 9p24.2p21.3. The clinical and cytogenetic findings in our and other similar patients are compared.

Reciprocal chromosome translocations involving short arm of chromosome 9 as a risk factor of unfavorable pregnancy outcomes after meiotic malsegregation 2:2.

PURPOSE: Genetic counseling of carriers with individual chromosome translocation requires information on how balanced reciprocal chromosome translocations (RCT) will segregate, what possible form of unbalanced embryo/fetus/child can occur, and the survival rates that have been observed in the particular families. We collected new empirical data and evaluated pedigrees of RCT carriers involving 9p in order to improve risk figures. MATERIAL AND METHODS: Empirical data on 241 pregnancies of 70 carriers were collected from 32 pedigrees of carriers of RCT at risk for a single 9p segment imbalance (RCT9p) from the literature and unpublished data. The probability rates of particular types of pathology have been calculated according to the method of Stengel-Rutkowski and Stene. Cytogenetic interpretation was based on GTG, RBG and FISH techniques. RESULTS: The probability rate for unbalanced offspring at birth for the whole group of pedigrees was calculated as 17.8+/-3% (33/185) (high risk). Considering the size of the imbalanced segment of 9p, the probability rates for RCT carriers with a breakpoint position at 9p22 at 9p13 and at 9p11.2 were estimated separately, and were found as 21.2+/-4.4% (18/85), 25+/-8.8% (6/24) and 11.8+/-3.7% (9/76), respectively. For unbalanced fetuses at 2nd prenatal diagnosis, we found the risk value as 57.9+/-11.3 % (11/19). The risk value for unkaryotyped stillbirths/early deaths of newborns and miscarriages were 5.4+/-1.7% (10/185) (medium risk) and 13+/-2.8% (rate 24/185) (high risk) respectively. CONCLUSIONS: Our results showed that the recurrence probability rates are different for particular categories of unfavorable pregnancy outcomes. How much they are dependent on the size of 9p chromosome segments taking part in the imbalance needs further studies based on a larger number of observations.

Trisomy of medial 15q as result of an analphoid supernumerary ring chromosome detected by CGH and FISH.

We report a 21-year-old patient with a de novo mosaic, analphoid ring of chromosome 15q22.2-->q24.1. The clinical features of this patient are mild and include tall stature, obesity, striae distensae in the hypogastrium, malocclusion and bilateral gynecomastia with scarce glandular tissue. M-FISH and FISH using a chromosome 15 painting probe indicated that the ring is of chromosome 15 origin. Further CGH analysis and FISH with the PML locus-specific probe demonstrated that the extra material derived from the medial part of the long arm of chromosome 15, including two bands, q22 and q23. Additionally, FISH with BAC probes specific for 15q allowed for a localization of the breakpoints at 15q22.2 and 15q24.1, distal to clones RP11-30M4 and RP11-500O23 respectively. We discuss the relationship between the patient's genotype and phenotype comparing it to reported cases of trisomy of medial 15q.

Richter syndrome first manifesting as cutaneous B-cell lymphoma clonally distinct from primary B-cell chronic lymphocytic leukaemia.

Richter syndrome (RS) is a transformation to high-grade non-Hodgkin lymphoma in patients with chronic lymphocytic leukaemia (CLL). RS may develop in lymph nodes or rarely extranodally. Skin localization of RS has been described in only a few cases. We present a 77-year-old woman who developed isolated diffuse large B-cell lymphoma (LBCL) in the skin of the nose without any other symptoms of RS. The LBCL in the skin was clonally distinct from the original bone marrow CLL cells. Moreover, LBCL cells were positive for LMP-1 segment of Epstein-Barr virus and overexpressed p53 protein. The patient was successfully treated with CHOP (cyclophosphamide, doxorubicin, vincristine and prednisone) and adjuvant local radiotherapy.

Extra structurally abnormal chromosomes (ESACs)--presentation of 10 new cases.

BACKGROUND: The goal of the presented studies as a retrospective reliability assessment of classical banding cytogenetic studies and of prognosing epicrises in a group of 14 cases, affected with additional marker chromosomes. MATERIAL AND METHODS: Having collected the study material from peripheral blood, by means of trophoblast biopsy or amniocentesis, cytogenetic preparations were obtained, allowing for pre- or postnatal evaluation of the karyotype. A panel of auxiliary cytogenetic techniques accompanied the routine CTG protocol. RESULTS: In a group of 6875 persons with recommendations to pre- or postnatal cytogenetic diagnostics, 14 (0.2%) cases of ESACs were diagnosed. In 5 cases of DA/DAPI(+) inv dup (15) as observed. A presence of polymorphic interstitial RHG(+) band was found within the marker chromosome. The measured size of that band allowed associating it with either the presence or the absence of pathological signs. In 9 cases of ESACs, DA/DAPI(-), the application of banding techniques (NOR and CBG) allowed to discover bisatellite heterochromatic ESACs in 6 cases (2 non-mosaic and 4 mosaic). In three other mosaic and non-satellite cases of ESACs, a 'genetic inactivity' of the marker chromosome was observed in one case, while a 'genetic activity' was ascertained in two cases. The 'activity' of marker chromosomes was studied by means of replication banding techniques. CONCLUSIONS: At the time of the outburst of molecular techniques, still up-to-date is the use of classical banding techniques and of the replication techniques, allowing DNA replication kinetics studies at the level of single band.

Application of FISH and Q-PCR techniques in breakpoint diagnostics in three cases of marker chromosomes derived from chromosome 15.

BACKGROUND: The goal of the study was a search for effective methods of diagnosing additional marker chromosomes. MATERIAL AND METHODS: Three cases of extra structurally abnormal chromosomes (ESACs) were diagnosed, the ESACs having been derived from chromosome 15 by cytogenetic techniques, the fluorescence in situ hybridisation (FISH) technique and the quantitative--polymerase chain reaction (Q-PCR). An application of a set of commercially available probes, specific for the 15q11.2-q12 regions (PWACR-Prader-Willi/Angelman Critical Region) allowed for a description of the breaking points. RESULTS: The presence of PWACR region was confirmed in one case and excluded in the other two. It was also attempted to apply the Q-PCR technique for a more accurate determination of the size of the region involved in chromosomal aberration, what would allow for a more reliable prognosing of the clinical outcome. In one of the patients, the breaking point was localized as distal to D15S144 locus, while it was proximal to D15S11 locus in the two remaining cases. CONCLUSIONS: The obtained results demonstrate a possibility of using the Q-PCR method in diagnosing unbalanced chromosome aberrations.

The quantitative PCR technique resolves ambiguities concerning a small rearrangement of human chromosome 6q12-13.

Karyotype analysis, performed on the basis of chromosome banding pattern, is a standard method used for identification of chromosomal aberrations, both numerical and structural. The application of classic cytogenetic techniques fails, however, to solve all diagnostic problems in certain types of chromosome aberrations. In this study, quantitative polymerase chain reaction technique (Q-PCR) application was applied to verify a cytogenetic diagnosis, which assumed that a difference observed in the banding pattern of homologous chromosome 6q12-13 region of a foetus had resulted from an inversion and/or duplication of the region in question. The obtained results indicate a possibility to use the Q-PCR method in the diagnostics of unbalanced chromosomal aberrations.

Molecular analysis of Y chromosome long arm structural instability in patients with gonadal dysfunction.

We have performed cytogenetic and molecular analyses of 45,X mosaics involving structurally abnormal Y chromosomes. Karyotypes were performed by standard cytogenetic methods and, in some cases, by fluorescence in situ hybridization, to distinguish monocentric and dicentric chromosomes. In addition, the deletions of Yq have been mapped using Southern blotting and polymerase chain reaction analysis. This paper provides additional information on the analysis of Y chromosome aberrations, and suggests that the stability of the Y chromosome in these instances is related to the site of the break point on Yq.

Umbilical and middle cerebral artery blood velocimetry: relationship with placental volume.

OBJECTIVE OF THE STUDY: To investigate the relationship between placental volume and the resistance of fetal arteries: umbilical and middle cerebral. MATERIAL AND METHODS: In 82 pregnant women with unaffected pregnancy between 14 and 40 weeks of gestation measurements of placental volume were made according to the parallel planimetric technique. Doppler examinations of umbilical and middle cerebral arteries were performed by means of a duplex system. RESULTS AND CONCLUSION: PI in the umbilical artery (PIUA) correlated significantly negatively with placental volume, whereas no significant correlation was found for the PI in the middle cerebral artery (PIMCA) and placental volume. Middle cerebral artery to umbilical artery PI ratio (PIMCA/PIUA ratio) showed significant positive correlation with placental volume. PIUA showed significant negative correlation with the relative volume of placenta (expressed as multiple of median for the actual gestational age) before 25 weeks of gestation and weak positive correlation with the relative volume of placenta after 25 weeks of gestation. PIMCA showed weak negative correlation with the relative volume of the placenta after 25 weeks of gestation and no correlation before 25 weeks. PIMCA/PIUA ratio showed significant positive correlation with relative placental volume before 25 weeks and significant negative correlation with relative placental volume after 25 weeks.

Prenatal screening for fetal chromosomal abnormalities using ear length and shape as an ultrasound marker.

INTRODUCTION: To determine the range of fetal ear length and ear shape variability in healthy fetuses and in fetuses with aneuploidies in the population of patients of our institution and to evaluate the clinical value of fetal ear length and shape assessment in screening for fetal aneuploidies. MATERIAL AND METHODS: Fetal ear length was determined in 210 healthy and 15 aneuploid fetuses of pregnant women undergoing targeted ultrasound examination. Fetal ear shape was assessed in the coronal section of the head and classified into one of three categories: flat, slightly protruding, markedly protruding and curved in 175 healthy and 14 aneuploid fetuses. RESULTS: Fetal ear length showed significant positive linear relationship with gestational age, which could be described by the following equation: Ear length (mm) = 0.968566 x gestational age (weeks) - 4.81629. Fetal ear shortening below 79% of expected length yielded the sensitivity of 26.7% in the detection of fetal aneuploidy at the false positive rate of 4.8%. Finding of markedly protruding and curved ears predicted fetal aneuploidy with the sensitivity of 28.6% at the false positive rate of 1.7%. The combination of fetal ear shortening below 74% of expected length or markedly protruding and curved ears resulted in the sensitivity of 33.3% and false positive rate of 3.33% in the detection of fetal aneuploidies. CONCLUSION: Fetal ear shortening has been confirmed as a marker of fetal aneuploidies. Abnormal fetal ear shape has been shown to constitute an additional marker of fetal aneuploidies, increasing the clinical value of fetal ear length measurements.

Utility of the triple test in the detection of abnormalities of the feto-placental unit.

INTRODUCTION: The objective of the study was to examine the relationship between the concentrations of maternal serum AFP, total beta-hCG and uE3 and the presence of various pathologies of the feto-placental unit. MATERIAL AND METHODS: The triple test was performed in the group of 882 pregnant women between the 14th and 21st week of gestation. The control group consisted of 863 pregnant women with unburdened obstetric anamnesis with an uneventful singleton pregnancy. The examined group consisted of 19 pregnant women with fetal aneuploidies and other pathologies of the feto-placental unit (oligohydramnios, vacuolization of the trophoblast). In the maternal sera concentrations of AFP, total beta-hCG and unconjugated estriol were determined. The risk of fetal trisomy 21 was calculated using DOWNS and PRISCA 3.0 software (cut-off 1: 295), Ulm Index was also calculated (cut-off 8.1). RESULTS: In the group of pregnant women with fetal chromosomal aneuploidies no significant changes of the examined parameters were established. In the cases of oligohydramnios AFP concentrations significantly increased, in the cases of fetal triploidy the concentrations of all examined parameters decreased. In the cases of vacuolization of the trophoblast concentration of the total beta-hCG increased. CONCLUSION: Oligohydramnios, fetal triploidy and vacuolization of the trophoblast caused significant changes of the concentrations AFP, total beta-hCG and unconjugated estriol in maternal sera.

[Neurological aspects of two patients with non-mosaic and mosaic polysomy of the X and Y chromosomes]. / Aspekty neurologiczne u dwóch chorych z niemozaikowa i mozaikowa polisomia chromosomów X i Y.

Clinical features were compared of a patient with the 48,XXYY karyotype and a case of 47,XXY/48,XXYY mosaicism. In the former patient tremor of the upper extremities of unclear aetiology was present. In both cases epilepsy was suspected. Similarly as in other cases of 48,XXYY karyotype the first patient had skeletal anomalies, abnormalities of dermatoglyphics and personality changes. These features are rarely found in Klinefelter syndrome. The differences in relation to the syndrome were less evident in the case of mosaicism 47,XXY/48,XXYY.

Analysis of triple test results in 27 cases of twin pregnancies.

The study comprises 889 pregnant women between 14 and 21 weeks of gestation. The control group consisted of 862 pregnant women with unburdened obstetric anamnesis with an uneventful singleton pregnancy. The examined group consisted of 27 pregnant women with uncomplicated twin pregnancy. In the sera of pregnant women AFP (Microparticle Enzyme Immunoassay AxSYM Abbott), total beta-hCG (Microparticle Enzyme Immunoassay AxSYM Abbott) and unconjugated estriol (Radioimmunoassay Amerlex-M. 2T Johnson & Johnson Ortho Clinical Diagnostics Ltd.) were determined. The risk of fetal trisomy 21 was calculated with the use of PRISCA 3.0 software, which corrected the MoM values for twin pregnancy. Ulm Index was also calculated. In the majority of twin pregnancies increased concentrations of AFP, total beta-hCG and uE3 in the range over 1.0 MoM was noted. In the group of women below 35 years of age with singleton pregnancies using PRISCA 3.0 software it approximated to 95%. For women older than 35 optimum index for fetal trisomy 21 risk calculation was Ulm Index with the specificity 93.8%. The specificity of AFP determination in the detection of fetal open NTD in singleton pregnancy was 99%. In the group of women with twin pregnancy the obtained specificity of 77.8% for PRISCA 3.0 software is low, a more advantageous way to calculate the risk of fetal trisomy 21 is Ulm Index with the specificity of 85.2%. The specificity of AFP determination as a screening for fetal open NTD in twin pregnancy was 96.3%.

[Inversion of chromosome 10 in a fetus with osteochondrodysplasia]. / Inwersja chromosomu 10 u plodu z osteochondrodyeplazja.

The case of thanatophoric dysplasia of the fetus was described. The disease was recognized by ultrasound. Karyotyping was successful after fetal blood sampling under continuous ultrasound guidance. An inversion of chromosome 10 was detected in the affected fetus. The confirmation of the thanatophoric dysplasia was done after termination of the pregnancy. The same inversion was detected in unaffected mother. Prenatal diagnosis was made in the next pregnancy. The result was an unaffected newborn delivered on term.

Cytogenetic and molecular identification of a de novo direct duplication of the long arm of chromosome 4(q21.3-->q31.3).

We report on a 3-year-old boy with moderate developmental retardation, microcephaly, and malformations of ears, lids, mouth, and thumbs. Cytogenetic analysis demonstrated a direct duplication of chromosome subregion 4(q21.3-->q31.3). Confirmation of this specific rearrangement was performed by fluorescent in situ hybridization (FISH) with a chromosome painting probe and by means of quantitative Southern hybridization with DNA probes localized within the chromosome 4 region presumed to be duplicated.

Two familial 9;17 translocations with variable effect on male carriers fertility.

Two similar, but not identical, familial reciprocal translocations are described. In the first family, four sterile males inherited reciprocal translocation t(9;17)(q11'3) of maternal origin. In the second family, with the male fertility not impaired, reciprocal t(9;17)(q11;q11) was observed in four members of the family.

De novo direct tandem duplication of the short arm of chromosome 7(p21.1-p14.2).

We report on a 6-year-old boy with moderate developmental retardation and unusual dermatoglyphics. Cytogenetic analysis demonstrated a duplication of chromosome sub-region 7p21.2-p14.2. Confirmation of the specific duplicated region was determined by quantitative Southern blotting by using a DNA fragment previously localized to the portion of chromosome 7 thought to be duplicated. This patient did not have the internal malformations seen with other dup(7p) patients.

A case of renal lymphangioma with a karyotype 45,X,-X,i dic(7q).

Cytogenetic analysis of a polycystic unilateral renal lymphangioma was performed by short-term culture and banding methods. The tumor's cells showed an isochromosome of the long arm of chromosome #7 and monosomy of X chromosome, whereas the peripheral lymphocytes stimulated with phytohemagglutinin showed a normal female karyotype, 46,XX. These karyotypic anomalies suggest that lymphangioma, although clinically benign, may have malignant potential.