G-banding Method Research Articles

Prevalence of Y Chromosome Microdeletions in Idiopathic Azoospermia Cases in Central Indian Men.

Genetic factor is important determinant of human male fertility, it is involved in 10-15% infertile males. Chromosome abnormalities and Y chromosome microdeletions are the main genetic causative factors for infertility. The frequency of male infertility & microdeletions in Y chromosome are also related to ethnic, geographical variations. In this study, we evaluated the prevalence of chromosomal abnormalities and microdeletions of Y chromosome in infertile azoospermia cases in central India to assess the geographical or population based variations. We have studied 160 non-obstructive azoospermia cases to find out frequency of chromosomal abnormalities and Y chromosome microdeletions of AZF locus. G-banding method was used for exclusion of chromosomal abnormalities. One hundred and forty eight azoospermic infertile men were screened using 12 sequence-tagged-sites (STS) primers of AZFa, AZFb, AZFc region and SRY gene (Yp) region by polymerase chain reactions. Out of 160 azoospermic infertile males, 12 (7.5%) confirmed chromosomal abnormalities and Klinefelter's syndrome was predominantly cause of azoospermia. Of the 148 infertile males, 19 (12.8%) were shown microdeletions in different AZF regions. Deletions in AZFa region were 2.02% and 3.37% was in AZFb whereas high frequencies of deletions (6.08%) in AZFc were recorded in azoospermic males. In two azoospermic males were shown microdeletions in AZFb+c loci. The prevalence of Y chromosome microdeletions in azoospermic men was 12.8% in this geographical region. Klinefelter's syndrome is important cause in male infertility. So, the screening of Y microdeletions is essential.

Abnormalities in spontaneous abortions detected by G-banding and chromosomal microarray analysis (CMA) at a national reference laboratory.

BackgroundCytogenetic evaluation of products of conception (POC) for chromosomal abnormalities is central to determining the cause of pregnancy loss. We compared the test success rates in various specimen types and the frequencies of chromosomal abnormalities detected by G-banding analysis with those found by Oligo-SNP chromosomal microarray analysis (CMA). We evaluated the benefit of CMA testing in cases of failed culture growth.MethodsConventional cytogenetic results of 5457 consecutive POC specimens were reviewed and categorized as placental villi, fetal parts, and unspecified POC tissue. The CMA was performed on 268 cases. Of those, 32 cases had concurrent G-banding results. The remaining 236 cases included 107 cases with culture failure and 129 cases evaluated by CMA alone.ResultsThe overall POC culture success rate was 75%, with the lowest for fetal parts (37.4%) and the highest for placental villi (81%). The abnormality rate was 58% for placental villi, but only 25% for fetal parts. Of the abnormalities detected, the most common were aneuploidies, including trisomy 16, triploidy, monosomy X, trisomy 22, trisomy 21 and trisomy 15, while the least encountered aneuploidies were trisomy 1, trisomy 19 and monosomies (except monosomy 21). Overall, POC specimens studied by CMA were successful in 89.6% of cases and yielded a 44.6% abnormality rate.ConclusionsPlacental villi yielded higher rates of culture success and a higher percentage of abnormal karyotypes than did other specimen types. The Oligo-SNP CMA method has demonstrated a viable alternative to the G-banding method in view of its advantages in detection of submicroscopic genomic aberrations, shorter turnaround time due to elimination of time required for culture and a higher test success rate.

Enlargement of bilateral kidneys by infiltration of leukemic cells in acute lymphoblastic leukemia at relapse after allogeneic hematopoietic stem cell transplantation

A 65-year-old man was admitted to our hospital because of multiple bullous lesions in the right leg in August 2011. Laboratory examinations showed pancytopenia: WBC 2,500/ll, Hb 12.3 g/dl, and Plt 10.8 9 10/ll, with a few abnormal circulating cells (3 %). Bone marrow aspiration showed a hypercellular marrow with an increase of abnormal blasts. Flow cytometric study of the blasts showed positivity for CD19, CD34, HLA-DR, and KORSA, and negativity for CD10 and CD20. Karyotype analysis using the G-banding method indicated complex abnormalities. Philadelphia chromosome was negative in fluorescence in situ hybridization (FISH) analysis. On the basis of these results, the patient was diagnosed with Philadelphia chromosome-negative, early precursor B cell acute lymphoblastic leukemia (B-ALL). After the diagnosis of B-ALL, induction chemotherapy was started. In the period of neutropenia, he developed sepsis, and wholebody computed tomography (CT) showed diffuse pulmonary infiltrative shadows, mild splenomegaly, and normal kidney size (Fig. 1a). This sepsis was controlled, and the patient achieved complete remission (CR). After 2 courses of consolidation chemotherapy, he received bone marrow transplantation from an HLA/ABO-matched unrelated donor. Engraftment was successful, and acute GVHD was not seen. Unfortunately, he showed relapse in the bone marrow at day 92. At relapse, whole-body CT indicated bilateral renal enlargement without mass formation (Fig. 1b). He received several courses of chemotherapy again, but died due to disease progression 7 months after allogeneic transplantation. Autopsy of the right kidney was performed. Pathological examination showed that numerous abnormal lymphoid leukemic cells had infiltrated into the renal interstitial tissue, although the glomeruli were intact (Fig. 2).

Tetraploidy in the era of molecular karyotyping – What we need to remember

Tetraploidy is a condition in which there are four complete sets of chromosomes in a single cell. In humans, this would be 92 pairs of chromosomes per cell. A great majority of pregnancies with a tetraploid fetus end in miscarriage, or if the pregnancy goes to full term, the infant dies shortly after birth. Longer surveillance is rarely described. The only method for confirming tetraploidy is karyotyping, in many cases using classical G-banding methods. In this paper we would like to present another living individual with tetraploidy and to revise the syndrome in the light of its diagnostics in the era of molecular karyotyping, with array Comparative Genomic Hybridization (aCGH, arrayCGH).

A new form of the mole vole Ellobius tancrei Blasius, 1884 (Mammalia, Rodentia) with the lowestchromosome number.

The subterranean mole vole, Ellobius tancrei, with aspecific variability in autosomes (2n = 31–54) and unusual sex chromosomes (XX in males and females), represents an amazing model for studying the role of chromosome changes in speciation. New materials from the upper reaches of the Surkhob River in the Pamiro-Alay mountains resulted in the discovery of a new form with 2n = 30. The application of Zoo-FISH and G-banding methods allowed the detection of 13 pairs of autosomes as Robertsonian metacentrics originated after fusions of acrocentrics of an assumed ancestral karyotype of Ellobius tancrei with 2n = 54. The sex chromosomes (XX, in both sexes) and one pair of acrocentric autosomes are the only acrocentrics in this karyotype, and the set with 2n = 30 possesses the lowest possible chromosome number among populations of Ellobius tancrei.

High prevalence of genetic abnormalities in Middle Eastern patients with idiopathic non-obstructive azoospermia

Our objective is to detect the frequency and types of major genetic abnormalities of idiopathic non-obstructive azoospermia (NOA) to give appropriate genetic counseling before assisted reproductive techniques (ART) in Middle East and to compare the frequencies with other regions of the world. A total of 880 Middle Eastern patients with NOA were recruited in this multicenter study for genetic evaluation prior to use of ART. Karyotyping was performed on peripheral blood lymphocytes according to standard G-banding methods, polymerase chain reaction (PCR) was performed to screen the microdeletions in the AZF region of the Y chromosome. The present study shows that the total prevalence of genetic abnormalities is 28.41 %, including 184 patients (20.91 %) with chromosome disorder and 66 patients (7.5 %) with Y chromosome microdeletions. The most prevalent chromosome abnormality is Klinefelter's syndrome, which includes 161 patients (18.3 %), 7 patients had XX reversal male sex (0.8 %), 2 patients had 47XYY (0.23 %) and 2 patients had 45XO/46XY (0.23 %). Structural abnormalities occurred in 12 patients (1.36 %). The high prevalence of genetic abnormalities (28.41 %) in our study strongly suggests the need for routine genetic testing and counseling prior to assisted reproduction in such population with idiopathic infertility, as a result may help determine the prognosis, as well as the choice of ART. Moreover it allows specific pre-implantation genetic testing to minimize the risk of transmitting genetic defects to offspring.

Diagnóstico de aneuplodias en células en interfase de líquido amniótico mediante hibridación in situ fluorescente. Concordancia con el cariotipo

BackgroundThe application of fluorescence in situ hybridization (FISH) techniques in the last few years has led to the prenatal diagnosis of aneuploidies. The objective of this investigation was a descriptive analysis of amniotic fluids processed in the laboratory using FISH and the agreement with the karyotype. Material and methodsA total of 821 amniotic fluid samples (January 2009 to December 2010) at gestational ages 13 to 36 weeks, from Fetal Medicine Unit for prenatal testing for aneuploidies (Aneuvysion kit) with centromeric probes for chromosomes X,Y and 18, and locus specific for chromosomes 13 and 21. The study was complemented by the karyotype by G-banding method. ResultsOf the 821 samples, 776 (94.52%) were normal and 45 (5.48%) had aneuploidy: in 22 cases (48.88%) the chromosomal sex of the foetus was male, in the remaining 23 (51.12%) female. The most common chromosomal abnormality detected was trisomy 21 (19 cases in males and 11 cases in female foetuses), the lowest performance was the trisomy 13 (2 cases) representing 0.24% of total processed amniotic fluids. In all cases, the concordance with the karyotype was 100%. ConclusionsThe study of aneuploidy by FISH of interphase nuclei present in the amniotic fluid enables rapid prenatal diagnosis of major chromosomal abnormalities, trisomy 21 was more frequently detected. FISH results in 100% agreed with the karyotype, the gold standard in prenatal diagnosis of chromosomal abnormalities

Abstract 1422: High FLT3 mRNA expression without FLT3-ITD in infants with acute myeloid leukemia

Abstract Introduction: When compared to older patients, infants with acute leukemia exhibit high frequency of MLL gene rearrangement (MLL-R), and are known to have higher vulnerability to intensive cytotoxic therapy, such as hematopoietic stem cell transplantation (HSCT). In contrast to acute lymphoblastic leukemia (ALL) in infants, there have been a few reports on acute myeloid leukemia (AML) in infants including our group (A. Shimada et al, ASH 2011). We and other group reported that MLL-R had the clinical impact in infant ALL, however, not in infant AML. Infant ALL with MLL-R expressed high FLT3 mRNA, on the other hand, the importance of FLT3 mRNA remains to be obscure in infant AML. Patients: Forty four infants with AML, aged less than 1 year at diagnosis, registered in JPLSG AML-05 from 2006-2010. Patients with Down syndrome were excluded. The most frequent FAB classification type was M7 (18/44, 40.9%), followed by M5 (15/44, 34.1%), and M4 (4/44, 9.0%). Total 16 of 44 (36.4%) infant AML patients had MLL-R, 9 patients were detected by G-banding method, and other 7 patients were by FISH analysis or RT-PCR. There was not t(8;21), t(15;17) except for one inv(16) patient. Methods: FLT3 mRNA expression and internal tandem duplication (ITD) were analyzed in the first diagnostic bone marrow samples by RQ-PCR and PCR. Results: No patients had FLT3-ITD, however, higher FLT3 mRNA expression was found in MLL-R patients (n=16) (median 623,150 copies/μgRNA) compared with MLL-wild type (WT) (n=28) (median 25,105) (p<0.005). This phenomenon was also observed in 12-23 months AML patients. Discussion: High FLT3 mRNA expression was reported in infant ALL with MLL-R and it can be a molecular target for FLT3 inhibitor. On the other hand, there was no report in infant AML. Our previous data suggested that SCT had the limited efficacy for infant AML, FLT3 inhibitor might be considered to be used for infant AML with MLL-R in future. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1422. doi:1538-7445.AM2012-1422

Mixed phenotype acute leukemia of T/myeloid type with a prominent cellular heterogeneity and unique karyotypic aberration 45,XY, dic(11;17)

A 26-yr-old male patient with mixed phenotype acute leukemia of T/myeloid type with prominent leukemic cell heterogeneity, and the presence of a so far unreported karyotype aberration in this type of acute leukemia 45,XY, dic(11;17)(11qter→11p11.2::17p11.2→17qter) is presented. Flow immunocytometry was performed by direct multicolor immunofluorescent technique on bone marrow aspirates. Cytogenetic analyses were performed using G-banding method by direct preparation of unstimulated bone marrow cells and following 24 hours of culture in RPMI 1540 culture medium with 25% fetal calf serum at 37°C RESULTS: The flow immunocytometry of bone marrow nucleated cells revealed the existance of three distinct blast cell populations with overlapping immunophenotypes. Predominant blast cell population had an early myeloid phenotype and aberrant expression of CD7 antigen (HLA-DR(+), CD34(+), anti-MPO(+), CD117(+), CD33(+), CD13(+), CD7(+low), cyCD3(-), TdT(-)). The other two blast cell populations, smaller in cell diameter and less sizable in cell proportion, both shared the T-lymphoid features. The patient was treated with ADE protocol (etoposide, cytarabine and doxorubicine). A complete remission was achieved and lasted 5 months. A case of MPAL with complex biological features, 45,XY, dic(11;17)(11qter→11p11.2::17p11.2→17qter) karyotype and an aggressive, therapy-resistant clinical course, is presented.

Karyotype of NIH, C57BL/6 and Razi strains of laboratory mice (Mus musculus)

The laboratory mouse is recognized as the pre-eminent model for genetic research. Awareness of chromosomal patterns of experimental animals increases their value for a variety of different fields of study. We aimed to study mitotic chromosome preparations from NIH, C57BL/6 and Razi strains of mice, which are outbred, inbred and partially inbred laboratory mice respectively. Bone marrow cells were prepared from 36 male and female mice, 12 from each strain, and stained by use of Giemsa staining and G-banding methods. Karyotyping of the samples showed that there was no difference in chromosomal numbers among the three mice strains, also the metaphase preparations of their diploid cells contained 40 chromosomes (2n = 40) and all chromosomes were telocentric. However, some differences in band tonality and the size of chromosomes were seen.

Pleomorphic type of malignant fibrous histiocytoma with myxoid stroma of the vulva in a young woman

Malignant fibrous histiocytoma (MFH) of the vulva is extremely rare; to date, there have been nine case reports. Almost all of these cases involved middle-aged women. We encountered a 21-year-old woman with a 4.5 × 2.6 cm superficial, localized, exophytic tumor of the right vulva. Microscopic findings on punch biopsy of the tumor initially suggested a vulvar sarcoma. The patient underwent wide local excision of the vulva. The extirpated specimen demonstrated the pleomorphic type of MFH with myxoid stroma of the vulva. The clinical stage was found to be IIB, based on the American Joint Committee on Cancer staging system. Chromosomal analysis of the tumor using the conventional G-band method was normal (46XX). This seems to be a very rare case of MFH of the vulva in a young woman. Physicians should therefore include MFH in the differential diagnosis of vulvar tumors, even though it is a rare disease.

Clinical Impact of Endoscopic Ultrasound-Guided Fine Needle Aspiration Biopsy In the Diagnosis of Patients with Lymphoma Who Have No Superficial Lymphadenopathies

AbstractAbstract 5083 Background:In patients with mediastinal and intra-abdominal lymphoma who do not have superficial lymphadenopathies, the diagnosis is usually made by surgical sampling with open thoracic surgery, laparotomy, mediastinoscopy, thoracoscopy, or laparoscopy. However, these surgical procedures are cumbersome and invasive; therefore, the treatment is often delayed, and doctors often hesitate to attempt the procedures for patients in poor performance status (PS). Recently, endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) has been used for the diagnosis of unknown mediastinal and abdominal lymphadenopathies or masses. With this procedure, pathological materials can be obtained through the gastrointestinal tract under real-time ultrasound imaging guidance. It is regarded as a minimally invasive, safe, and relatively accurate procedure. However, only a small amount of material can be sampled with this technique. Immunohistochemical, flow cytometric, and chromosomal analyses are essential for the diagnosis of lymphoma, but it is not clear whether these analyses can be assessed on samples obtained by EUS-FNA. Aim:To evaluate the yield of EUS-FNA for the diagnosis of lymphoma. Patients and Methods:Between June 2005 and December 2009, a total of 224 consecutive patients who were suspected to have lymphoma underwent EUS-FNA with a 19-gauge needle at our hospital. We reviewed the baseline characteristics of the patients; the histological diagnosis including immunohistochemical stainings, flow cytometric analysis (FCM), and chromosomal analysis (by the G-banding method) of the FNA samples; and the final diagnosis. EUS-FNA was performed in 241 lesions in the 224 patients. One hundred and forty-three patients (63.8%) underwent the procedure as outpatients. Ten in 224 patients had a poor PS (grade 4). Results:The locations of the lesions were mediastinal in 99 and intra-abdominal in 142 (inclusive of 9 splenic, 5 left adrenal, 2 pancreatic, and 1 right renal lesions). After all, 142 in 224 patients were finally diagnosed as having lymphoma. Among them, 138 in 142 patients (97.2%) were diagnosed by EUS-FNA, and detailed classification in accordance with the WHO system was also possible in 125 in 142 patients (88.0%) on the basis of comprehensive findings including immunohistochemical staining, FCM, FISH method, and chromosomal analysis. While, remaining 82 in 224 patients were diagnosed as having inflammatory diseases such as sarcoidosis, necrotizing lymphadenitis, tuberculosis, or other cancers. No serious complications occurred with the procedure. Conclusions:EUS-FNA is a safe and convenient procedure with high diagnostic value for the diagnosis of lymphoma. Therefore, this procedure should be initially considered to attempt for patients with mediastinal and intra-abdomnal lymphadenopathy or mass who do have no superficial lesions. Disclosures:No relevant conflicts of interest to declare.

Predictive Factors of Response and Survival Following Chemotherapy Treatment In Acute Leukemia with Complex Karyotype Aberrations

AbstractAbstract 4857 Background:Acute leukemia (AL) is a clonal hematopoietic stem cell disorder with diverse genetic abnormalities and various responses to treatment. AL was categorized into favorable, intermediate and adverse karyotype according to cytogenetic characteristics comprising different groups of patients with similar treatment response, relapse risk and overall survival. In particular, Patients with complex chromosome aberrations (CCA) harboring three or more acquired chromosome aberrations always have poor outcomes. It is necessary to investigate the clinical and laboratory features of AL patients with CCA, and to explore the underlying mechanisms of leukemogenesis in order to improve prognosis in these patients. Methods:83 AL patients with CAA were investigated and other 83 AL patients with none-complex chromosome aberrations (N-CCA) were regarded as control group from January 2005 to August 2008. The diagnosis of AL was based on morphological picture, immunophenotypical analysis, and cytochemical studies and FAB criteria were used for AL subtype classifications. Karyotypes were also analysed using conventional G-banding method. After patients had comfirmed diagnosis with AL, corresponding induction and consolidation chemotherapy in the two groups were given. Finally the treatment efficacy and prognosis were assessed. All the statistical analyses were performed using Kaplan-Meier. Results:There was no significant difference about sex, age and peripheral blood parameters before treatment between AL patients with CAA and the control group during the same period (p>0.05). CD34 was expressed in 72.3% of the AL patients with CCA, significantly higher than that in the control group (37.4%)(p<0.01). CD34+ AL patients with CCA showed a lower complete remission (CR) rate than in CD34- cases (p<0.05). 3-years EFS and OS were shorter in CD34+ patients than in CD34- cases. Using Kaplan-Meier analysis, the median survival time of AL patients with CCA was only 9 months, significantly shorter than the control group (21.5 months). There was no significant difference between AML and ALL patients with complex chromosome aberrations about CR rate and the survive time. Conclusion:The prognosis of patients with complex karyotype who received chemotherapy was extremely poor. In view of this, such patients should be considered as a unique entity and separated from those with a non-complex adverse cytogenetic change. They maybe benefit to some extent from allogeneic stem cell transplantation in younger patients according to some related reports. CD34 was associated with CAA while the underlying mechanisms between them were unknown. Disclosures:No relevant conflicts of interest to declare.

Evidence of chromosome fusion inGymnotus sylviusAlbert &amp; Fernandes-Matoli, 1999 (Teleostei: Gymnotiformes) detected by telomeric probes and R-banding

Gymnotus cf. carapo and Gymnotus sylvius are two fi sh species inhabiting the Upper Parana River Basin, presenting respectively 2n = 54 and 2n = 40 chromosomes. In the present cytogenetic analysis, R-banding and telomere-sequence hybridization were carried out in order to determine the possible relationship between the karyotipes of these two species. Incorporation bands (R-bands) obtained for the two species allowed the identifi cation of chromosome similarities, showing to be an usefull alternative to the G-banding methods, which fail in producing satisfying results in most of analyzed fi sh species. This approach, associated with the hybridiza- tion of telomeric sequences, permited to identify chromosomal rearrangements that could be used as indicators of karyotypic evolution within the group. In the present case, telomeric sequences were detected in the centro- meric region of two metacentric chromosome pairs of Gymnotus sylvius. The results obtained after hybridiza- tion with the telomere sequences, coupled with the chromosome homeologies detected by R-banding, showed that G. cf. carapo and G. sylvius should present a common ancestor, and this may also be corroborated by the similarities found in three chromosome pairs, that seem to have been conserved during the evolution of the two species. Based on the data here presented we propose that G. sylvius may have undergone a recent process of chromosome fusion that resulted in the diminution of its chromosome number.

Masked t(15;17) APL with the insertion of PML–RARα fusion gene in 4q21

Most cases of acute promyelocytic leukemia (APL) are characterized by the reciprocal translocation t(15;17); however, several complex variant translocations have also been reported. Here we report complex cytogenetic abnormalities without t(15;17) assayed by the G-banding method in a 62-year-old woman with the typical morphology and clinical features of APL. Based on spectral karyotyping and FISH analyses, we confirm the insertion of a cryptic chromosomal segment containing the PML/RARα fusion gene. The patient achieved complete remission after treatment with all-trans retinoic acid (ATRA) alone. Although the mechanism of this cryptic variant insertion is not known, we conclude that the insertion of PML–RARα fusion into 4q21 seems not to alter the effectiveness of treatment with ATRA.

Auer rod-like cytoplasmic inclusion bodies in nodal marginal zone lymphoma cells

A 58-year-old woman was referred to our hospital because of right axillary lymphadenopathy. A biopsy of the lymph node was performed. The lymph node section showed a diffuse proliferation of moderate-size atypical lymphoid cells with mild irregular-shaped nuclei and pale cytoplasm (Fig. 1). Immunohistochemical staining revealed that the atypical lymphoid cells were positive for CD19, CD20, and CD43, and negative for CD10, CD5, and cyclin D1. Cytogenetic analysis using the conventional G-banding method revealed normal karyotype. By FISH analysis, t(11;14)(q13;q32) was not detected. On the basis of these findings, the patient was diagnosed with nodal marginal zone lymphoma (MZL). No other surface lymph nodes were palpable at physical examination; however, some swollen lymph nodes adjacent to the sternum (Fig. 2a, arrow) were detected on computed tomography. In addition, fluorine-18 fluorodeoxyglucose (FDG) positron emission tomography (FDG-PET) revealed abnormal FDG accumulations at the swollen lymph nodes and in the sternum (Fig. 2b, arrows). Examination of bone marrow smears obtained from the sternal site showing FDG accumulation revealed many abnormal lymphoid cells; needleshaped inclusions that were morphologically similar to Auer rods were observed in the cell cytoplasm (Fig. 3a). Because the findings of bone marrow smears taken from the patient’s iliac bone were normal, we judged that these abnormal cells were lymphoma cells arising due to direct invasion to the sternum. Microscopic analysis revealed that the cytoplasmic inclusions had variable shapes. Although most lymphoma cells displayed multiple needle-like or splinter-shaped inclusions in their cytoplasm, some exhibited considerably thick inclusions (Fig. 3b). We did not find these inclusions in the lymph node biopsy specimen, which was prepared by fixing with formaldehyde and

Human leukocyte antigen-DR negative de novo acute myeloid leukemia with Philadelphia chromosome

It is well known that human leukocyte antigen (HLA)-DR is present in the majority of acute myeloid leukemia (AML) [1, 2]. However, acute promyelocytic leukemia (APL) with t(15;17)(q22;q12) usually lacks the HLA-DR expression as well as CD34 expression [2, 3]. Recently, such HLA-DR/CD34 phenotype has also been described in a subset of AML with cuplike nuclear invaginations (AML-cuplike) [4]. Cytogenetically or molecular genetically, AML-cuplike is characterized with normal karyotype and high frequency of FMS-like tyrosine kinase 3 (FLT3) gene internal tandem duplication (ITD) as well as nucleophosmin (NPM1) gene mutation [4, 5]. In this paper, we reported an extremely rare case of HLA-DR/CD34 AML, which was not categorized into either APL or AMLcuplike and showed Philadelphia chromosome (Ph) at diagnosis. A 64-year-old male was admitted to the hospital because of cellulitis of left leg in April 2008. He showed neither hepatosplenomegaly nor generalized lymph node swellings. There was no bleeding tendency. Laboratory findings were as follows: white blood cells 17.7 9 10/L, with 25% blasts and 20% basophils; hemoglobin, 14.0 g/dL; platelets, 129 9 10/L. Serum lactate dehydrogenase level and D-dimer was elevated to 386 IU/L and 50.2 lg/mL, respectively. Neutrophil alkaline phosphatase (NAP) score was within the normal range. Bone marrow showed hypercellularity with 39% blasts, 7% myelocytes, 4% metamyelocytes, 13% mature neutrophils, 1% eosinophils, 25% basophils, 8% erythroblasts and 3% lymphocytes. Morphologically, blasts were medium to large, and had round or sometimes slightly indented nuclei with dispersed chromatin. Fine azurophilic granules were sometimes found in their scant basophilic cytoplasm, but typical Auer-bodies were not detected (Fig. 1). They were cytochemically positive to myeloperoxidase (MPO) staining. Therefore, he was morphologically diagnosed as having AML-M2 according to the French–American–British (FAB) classification. There were no characteristic cuplike nuclear invaginations suggesting a diagnosis of AML-cuplike on either Wright–Giemsa or MPO staining [6]. Flow cytometric immunophenotyping using the CD45 gating method showed that leukemic blasts were CD2, CD3, CD4, CD5, CD7, CD8, CD10, CD13, CD15, CD19, CD20, CD33, CD34, CD41, CD90, CD117 and HLA-DR, suggesting HLA-DR AML (Fig. 2). More than 85% of these cells also lacked the CD56 expression. Chromosomal analysis of bone marrow using the conventional G-banding method showed t(9;22)(q34;q11) in 14 of the 20 metaphases analyzed in addition to normal variance of inv(9)(p12q13) (Fig. 3). Fluorescence in situ hybridization (FISH) also showed BCR/ABL fusion signal in 92% of T. Inaba (&) N. Fujita Department of Infection Control and Laboratory Medicine, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto 602-8566, Japan e-mail: inaba178@koto.kpu-m.ac.jp

Antiapoptotic genes are overexpressed in the group of patients with locally advanced rectal adenocarcinomas who do not respond to neoadjuvant chemoradiotherapy

Tetraploidy is a condition in which there are four complete sets of chromosomes in a single cell. In humans, this would be 92 pairs of chromosomes per cell. A great majority of pregnancies with a tetraploid fetus end in miscarriage, or if the pregnancy goes to full term, the infant dies shortly after birth. Longer surveillance is rarely described. The only method for confirming tetraploidy is karyotyping, in many cases using classical G-banding methods. In this paper we would like to present another living individual with tetraploidy and to revise the syndrome in the light of its diagnostics in the era of molecular karyotyping, with array Comparative Genomic Hybridization (aCGH, arrayCGH).

A new complex translocation t(5;17;15)(q11;q21;q22) in acute promyelocytic leukemia

Acute promyelocytic leukemia (APL) is associated with the t(15;17)(q22;q21) translocation which causes the fusion of the retinoic acid alpha gene ( RARA) on 17q21 to the promyelocytic leukemia gene ( PML) on 15q22. The two chimeric genes, PML/ RARA and RARA/ PML, are thought to play a role in leukemogenesis. A small proportion of patients with APL have complex or simple variants of this translocation. We report the case of a 22-year-old woman with APL carrying a complex variant translocation t(5;17;15)(q11;q12;q22) confirmed by G-banding, reverse transcription polymerase chain reaction (RT-PCR), fluorescence in situ hybridization(FISH), and spectral karyotyping analysis (SKY). The patient achieved complete remission with all-trans retinoic acid treatment and chemotherapy. These results illustrate the usefulness of combined analysis consisting of G-banding, RT-PCR, FISH, and SKY methods to identify the PML/ RARA fusion gene in cases with variant t(15;17).

The results of cytogenetic analyses in prenatal diagnosis

G-banding and other classical cytogenetic methods are still in use, together with molecular cytogenetic techniques such as FISH (Fluorescence In Situ Hybridization) and SKY (Spectral Karyotyping). This retrospective study evaluated clinical data on individuaols seeking genetic counseling over a 15-year period (1992 - 2007) at the Medical Genetic Center, Child and Youth Health Care Institute of Vojvodina in Novi Sad. The study included 37.191 genetic counselings, and 20.607 prenatal analyses (amniocentesis and cordocentesis). Over a 15-year period (1992 - 2007) 17.937 amniotic fluid samples were analyzed and 274 abnormal karyotypes were found; out of 2.670 fetal blood samples, there were 78 abnormal karyotypes. During a 15-year period, prenatal diagnosis, using amniocentesis and/or cordocentesis, showed 352 fetuses with chromosomal aberrations. On average, over the past 15-year period, 8% of pregnancies were controlled with invasive prenatal procedures. The percentage has changed; in fact, it is increasing from year to year. In 1992, only 0.82% (N=139/17000) of pregnant women in Vojvodina underwent invasive prenatal procedures, and in 2006 the rate increased to 15.65% (N=2660/17000). It is necessary to improve and promote the possibilities of genetic counseling and invasive prenatal diagnosis in order to prevent the occurrence of chromosomal aberrations and other genetic diseases.