Karyotype Research Articles

Correlative analysis between congenital heart diseases and Down's syndrome

Objective To investigate the association between Down's syndrome(DS) and congenital heart di-seases(CHD). Methods A total of 575 cases with DS from Jan.1997 to Mar.2013 in Children's Hospital of Chongqing Medical University were recruited.Retrospective study was conducted to analyze the prevalence and types of CHD in DS children, the relationship between the karyotype of DS and the types of CHD, and pulmonary hypertension(PH) and operation treatment. Results There were 370 cases(64.35%, 370/575 cases) with CHD in 575 cases with DS.Among the 370 cases of CHD, 322 cases(87.03%) were septal defects.In which, 57 cases(15.41%) were atrial septal defects, 36 cases(9.72%) were ventricular septal defects, 12 cases(3.24%) were atrioventricular septal defects, and 157 cases(47.30%) were complex septal defects.Forty-eight cases(12.97%, 48/370 cases) were non-septal defect types of CHD(including patent ductus arteriosus, tetralogy of Fallot, double outlet right ventricle, pulmonary atresia, and so on). There was no statistical significance between the karyotype of DS and the types of CHD.There were 246 cases(66.49%, 246/370 cases) with PH.Seventy cases(18.92%, 70/370 cases) had interventional or surgical operations.All of them had descending pulmonary artery pressure after operation.Forty cases had other malformations such as gastrointestinal tract malformation, polydactylism / polydactyly, visual impairment, and so on. Conclusions The most common type of CHD with DS was atrial septal defect, and the second one was ventricular septal defect.There was no relationship between the karyotype of DS and the types of CHD.The patients with CHD in DS were prone to develop PH.So the comprehensive treatment plan should be developed as early as possible. Key words: Down's syndrome; Congenital heart diseases; Pulmonary hypertension

Jumping translocations of 1q12 in multiple myeloma: a novel mechanism for deletion of 17p in cytogenetically defined high-risk disease

AbstractMultiple myeloma (MM) is a B-cell malignancy driven in part by increasing copy number alterations (CNAs) during disease progression. Prognostically significant CNAs accumulate during clonal evolution and include gains of 1q21 and deletions of 17p, among others. Unfortunately, the mechanisms underlying the accumulation of CNAs and resulting subclonal heterogeneity in high-risk MM are poorly understood. To investigate the impact of jumping translocations of 1q12 (JT1q12) on receptor chromosomes (RCs) and subsequent clonal evolution, we analyzed specimens from 86 patients selected for unbalanced 1q12 aberrations by G-banding. Utilizing spectral karyotyping and locus-specific fluorescence in situ hybridization, we identified 10 patients with unexpected focal amplifications of an RC that subsequently translocated as part of a sequential JT1q12 to one or more additional RCs. Four patients exhibited amplification and translocation of 8q24 (MYC), 3 showed amplification of 16q11, and 1 each displayed amplification of 18q21.3 (BCL2), 18q23, or 4p16 (FGFR3). Unexpectedly, in 6 of 14 patients with the combination of the t(4;14) and deletion of 17p, we identified the loss of 17p as resulting from a JT1q12. Here, we provide evidence that the JT1q12 is a mechanism for the simultaneous gain of 1q21 and deletion of 17p in cytogenetically defined high-risk disease.

Feasibility of rapid prenatal diagnosis in advanced maternal age women

Objective To analyze the feasibility of rapid prenatal diagnosis in the advanced maternal age women with or without positive serologic screening results.Methods We conducted a retrospective study of the women who underwent a mid-trimester amniocentesis in Peking University First Hospital from January 1,2001 to December 31,2012.Maternal age,indication for invasive prenatal diagnosis,karyotyping and pregnancy outcome were documented.Using a young population with high risk in serologic screening (S) as the standard,chromosome abnormalities in the advanced maternal age (A) group and the advanced maternal age with high risk in serologic screening (A+S) group were compared with the S group.Chromosome abnormalities were divided into detectable (D) and undetectable (U) during rapid prenatal diagnosis.Results Of 9 606 cases,222 (2.3％,222/9 606) cases with chromosome abnormalities were detected,23.0％ (51/222) of which were undetectable by rapid prenatal diagnosis.The detection rate of detectable chromosome abnormalities was 1.8％ (57/3 177) in group A,1.4％(13/925) in group A+S,and 1.8％(57/3 250) in group S (x2=0.662,P＞0.05).The rate of undetectable chromosome abnormalities was 0.5％ (15/3 177) in group A,0.3％ (3/925) in group A+S,and 0.5％ (16/3 250) in group S (x2=0.452,P＞0.05).The most common indications for undetectable chromosome abnormalities in the young population were abnormal history of pregnancy,abnormal family history and chromosome abnormality history (16.4％,9/55),and abnormal ultrasound in the advanced maternal age population (4.4％,3/68).Conclusions The performance of rapid prenatal diagnosis in the advanced maternal age population with or without high risk in screening without abnormal findings in ultrasound,was similar to the young population with high risk in screening.Fluorescent in situ hybridization may be integrated into the strategy of prenatal diagnosis for this group of women. Key words: Prenatal diagnosis; Amniocentesis; Karyotyping; In situ hybridization, fluorescence; Pregnancy trimester, second

Clinical feature and cytogenetic analysis of 80 patients with acute monocytic leukemia

To explore the relationship between cytogenetic and clinical features and prognosis for patients with acute monocytic leukemia (M5 type). Chromosome samples were prepared by direct culture of bone marrow for 24 hours. Karyotypes of 80 patients with M5 were analyzed by R banding. Rearrangement of MLL gene and deletion of P53 gene were detected by fluorescence in situ hybridization (FISH). Forty-three patients (53.75%) were found to have abnormal karyotypes, which included 23 patients with abnormalities of chromosome 11 and 20 with other chromosomal abnormalities. Twenty-four patients had MLL gene rearrangements and 17 had P53 gene deletion. 11q23 has been the most common chromosomal abnormality among patients with M5, which is associated with poor prognosis. The frequency of P53 gene deletion in patients with genetic abnormalities were significantly higher than those with normal karyotypes (P < 0.05).

Spectral karyotyping: an unique technique for the detection of complex genomic rearrangements in leukemia.

Spectral karyotyping (SKY) is a novel cytogenetic technique, has been developed to unambiguously display and identify all 24 humans chromosomes at one time without a priori knowledge of any abnormalities involved. SKY can discern the aberrations that can't be detected very well by conventional banding technique and Fluorescent in situ hybridization (FISH). So SKY is hyper accurate, hypersensitive, and hyper intuitionist. We will review the elements and application of SKY in leukemia.

Ultrasound soft markers and fetal cardiac structural assessment at 11-14 weeks

To evaluate the detection rate of congenital heart defect (CHD) during the first trimester screening for chromosomal abnormalities, the role of ultrasound soft markers including increased nuchal translucency (NT), tricuspid regurgitation (TR) and abnormal ductus venosus (DV) flow in the screening for cardiac anomalies was also investigated. From January 2009 to January 2012, 4 673 fetuses were scanned at 11-14 weeks at Department of Fetal Medicine, the First Affiliated Hospital of Jinan University. The ultrasound findings and follow up outcomes were recorded, False-positive rate of different first-trimester ultrasound markers for the detection of CHD was calculated, sensitivity of the markers for all major CHD was calculated as well. There was a significant association between major CHD and first trimester ultrasound markers. (1) Overall findings:among the 4 673 fetuses, 31 fetuses were diagnosed CHD prenatally, 17, 12 and 2 of which were detected in the first, second and third trimester, respectively. In 22 of the 31 CHD cases, invasive procedure was performed, fetal karyotype was abnormal in 12 cases, including triosmy 21 (5 cases), trisomy 18 (2 cases), trisomy 13 (2 cases), Turner syndrome (2 cases) and pericentric inversion of chromosome 9 (1 cases). (2) NT measurement and prenatal detected CHD: in 4 673 cases, NT measurement between 95(th)-99(th) percentile were present in 206 (4.41%) , 5 cases were diagnosed CHD prenatally, in 4 of 5 cases were detected in first trimester; NT measurement <95(th) percentile were present in 4 430 (94.80%) , 16 cases were diagnosed CHD prenatally, in 5 of 16 cases were detected in first trimester; NT measurement>99(th) percentile (>3.5 mm) were present in 37 (0.79%, 37/4 673) , 10 cases were diagnosed CHD prenatally, in 8 of 10 cases were detected in first trimester. (3) TR and inverted a-wave at the DV and prenatal detected CHD: among 4 673 cases, TR or inverted a-wave at the DV were present in 51 (1.09%) , 98 (2.10%) respectively. TR was present in 8 of 31 CHD cases, inverted a-wave at the DV was present in 7 of 31 CHD cases. (4) Sensitivity of different first trimester ultrasound markers for detection of major CHD cases:in 31 CHD cases diagnosed prenatally, 23 cased were defined as major CHD. Sensitivity of at least one of the ultrasound markers, NT measurement between 95(th)-99(th) percentile, >99(th) percentile( >3.5 mm), TR or inverted a-wave at the DV for detection of major CHD cases was 74% (17/23), 22% (5/23), 39% (9/23), 35% (8/23), 30% (7/23) , respectively. (5) Specificity of different first trimester ultrasound markers for detection of CHD cases:specificity of NT measurement between 95(th)-99(th) percentile, >99(th) percentile (>3.5 mm), TR or inverted a-wave at the DV for detection of major CHD cases was 4.30% (201/4 673),0.58% (27/4 673),0.92% (43/4 673), 1.94% (91/4 673). Routine first trimester soft markers for chromosomal abnormalities screening combined with cardiac assessment can detect quite a number of major heart defects. Increased NT, TR and abnormal DV flow can be important indicators for echocardiography, which is favorable to early prenatal diagnosis of CHD.

Tandem triplication of the BCL2 gene in CD5-positive intravascular large B cell lymphoma with bone marrow involvement

Dear Editor, Intravascular large B cell lymphoma (IVLBCL) is a rare subtype of diffuse large B cell lymphoma (DLBCL) characterized by the selective growth of lymphoma cells within the lumina of vessels [1]. Cytogenetically, the majority of IVLBCL cases demonstrated complex karyotypes with multiple numerical and structural changes, and the most frequent duplicated regions were shown to be the 11q13→qter region and all of chromosome 18 [2, 3]. On the other hand, triplications of chromosome segments other than 1q have been rarely found in hematological malignancies [4–8]. Here, we describe an unusual case of IVLBCL with a tandem triplication of the BCL2 gene at 18q21. An 82 year-old female was admitted because of fever. Computed tomography scans revealed hepatosplenomegaly without lymphadenopathy. Peripheral blood (PB) examination showed hemoglobin 105 g/L, platelets 52×10/L, and leukocytes 10.1× 10/Lwith 15% abnormal lymphocytes. Serum levels of lactate dehydrogenase (LDH) and soluble interleukin-2 receptor were 6,382 IU/L (normal range, 115~217) and 8,429 U/mL (124~ 466), respectively. Bone marrow (BM) was normocellular with 42.4 % abnormal lymphocytes positive for CD5, CD19, CD20, and HLA-DR (Fig. 1a). Pathological examination of the BM revealed a focal and sinusoidal growth pattern of large lymphoid cells, indicating a diagnosis of IVLBCL (Fig. 1b). Immunohistochemistry demonstrated that they were positive for CD5, CD20, BCL2, and MUM1, but negative for CD10 and BCL6 (Fig. 1c, d). The patient received R-CHOP therapy and achieved hematological complete remission. However, 12 months later, she relapsed and died of progressive disease. G banding and spectral karyotyping of BM cells showed 47,XX,del (2)(q?),+del(3)(p?),der(10)t(10;20)(q24;?), dup(17)(q21q23),dup(18)(q11.2q23) [20] (Fig. 1e). Unexpectedly, fluorescence in situ hybridization (FISH) detected three BCL2 signals on the dup(18)(q11.2q23), indicating a tandem triplication of BCL2 (Fig. 1f). Namely, the short 18q fragment containing BCL2 attached to the dup(18). FISH on interphase nuclei confirmed four BCL2 signals. According to the Mitelman Database, nine cases of hematological malignancies with duplication/triplication of 18q (duplication, eight; triplication, one) in the stem-lines have been described [4, 9, 10]. All cases were diagnosed with B lymphoid malignancies. The duplicated regions in two cases of acute lymphoblastic leukemia did not contain 18q21, whereas those in seven cases of B cell lymphomas (DLBCL, four; IVLBCL, one; Burkitt lymphoma, one; mantle cell lymphoma, one) retained 18q21. Thus, duplication/triplication of 18q including 18q21 seems to be specifically associated with B cell lymphomas and suggests the significance of BCL2 in their pathogenesis. Among these, one case with trp(18)(?q21q23) exhibited a tandem triplication of BCL2 [2]. This 62-year-old male was diagnosed as having IVLBCL with BM/PB involvement, thrombocytopenia (34×10/L), and high LDH (2,266 U/L). Lymphoma cells were positive for K. Yamamoto (*) :A. Okamura :K. Yakushijin :H. Matsuoka : H. Minami Division of Medical Oncology/Hematology, Department of Medicine, Graduate School of Medicine, Kobe University, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan e-mail: kyamamo@med.kobe-u.ac.jp

Ewing sarcoma of the small bowel: a study of seven cases, including one with the uncommonly reported EWSR1–FEV translocation

Primary Ewing sarcoma of the ileum has rarely been documented. Little is known about its pathogenesis and clinical implications, and it would be helpful to identify novel molecular markers. EWSR1-FEV translocation is exceedingly rare in Ewing sarcoma, as FEV expression is restricted to prostate, brain and serotonin neuroendocrine cells (NE) and related tumours. Paraffin sections or snap-frozen material were used in this investigation. Tumours were investigated by means of immunohistochemistry, RT-PCR (EWSR1-FLI1, EWSR1-ERG and EWSR1-FEV transcripts), FISH analysis (EWSR1 break-apart and specific EWSR1-FEV translocation) and spectral karyotyping (SKY). Ten ileal neuroendocrine tumours (INET) made up the control group for EWSR1-FEV translocation. Among 445 Ewing sarcomas cases spanning a period of 20years, seven (1.6%) arose in the ileum. All tumours were immunoreactive for synaptophysin, CD99, FLI1 and vimentin. FISH identified EWSR1 rearrangement in all cases, with EWSR1-FLI1 transcripts being detected in all but one tumour showing the uncommon EWSR1-FEV rearrangement, with SKY, RT-PCR and FISH confirmation. The mean survival of EWSR1-FLI1 patients was 14months, whereas the EWSR1-FEV patient was alive after 15years despite several recurrences controlled by surgery alone. No INET showed EWSR1 translocation. Most primary Ewing sarcomas of the ileum show the common EWSR1-FLI1 translocation, but EWSR1-FEV could be specific for tumours arising in the ileum and showing better prognosis.

Mechanisms of chromosomal instability in melanoma

A systems biology approach was applied to investigate the mechanisms of chromosomal instability in melanoma cell lines. Chromosomal instability was quantified using array comparative genomic hybridization to identify somatic copy number alterations (deletions and duplications). Primary human melanocytes displayed an average of 8.5 alterations per cell primarily representing known polymorphisms. Melanoma cell lines displayed 25 to 131 alterations per cell, with an average of 68, indicative of chromosomal instability. Copy number alterations included approximately equal numbers of deletions and duplications with greater numbers of hemizygous (-1,+1) alterations than homozygous (-2,+2). Melanoma oncogenes, such as BRAF and MITF, and tumor suppressor genes, such as CDKN2A/B and PTEN, were included in these alterations. Duplications and deletions were functional as there were significant correlations between DNA copy number and mRNA expression for these genes. Spectral karyotype analysis of three lines confirmed extensive chromosomal instability with polyploidy, aneuploidy, deletions, duplications, and chromosome rearrangements. Bioinformatic analysis identified a signature of gene expression that was correlated with chromosomal instability but this signature provided no clues to the mechanisms of instability. The signature failed to generate a significant (P = 0.105) prediction of melanoma progression in a separate dataset. Chromosomal instability was not correlated with elements of DNA damage response (DDR) such as radiosensitivity, nucleotide excision repair, expression of the DDR biomarkers γH2AX and P-CHEK2, nor G1 or G2 checkpoint function. Chromosomal instability in melanoma cell lines appears to influence gene function but it is not simply explained by alterations in the system of DDR.

Flow cytometric chromosome sorting from diploid progenitors of bread wheat, T. urartu, Ae. speltoides and Ae. tauschii

Chromosomes 5A (u) , 5S and 5D can be isolated from wild progenitors, providing a chromosome-based approach to develop tools for breeding and to study the genome evolution of wheat. The three subgenomes of hexaploid bread wheat originated from Triticum urartu (A(u)A(u)), from a species similar to Aegilops speltoides (SS) (progenitor of the B genome), and from Ae. tauschii (DD). Earlier studies indicated the potential of chromosome genomics to assist gene transfer from wild relatives of wheat and discover novel genes for wheat improvement. This study evaluates the potential of flow cytometric chromosome sorting in the diploid progenitors of bread wheat. Flow karyotypes obtained by analysing DAPI-stained chromosomes were characterized and the contents of the chromosome peaks were determined. FISH analysis with repetitive DNA probes proved that chromosomes 5A(u), 5S and 5D could be sorted with purities of 78-90 %, while the remaining chromosomes could be sorted in groups of three. Twenty-five conserved orthologous set (COS) markers covering wheat homoeologous chromosome groups 1-7 were used for PCR with DNA amplified from flow-sorted chromosomes and genomic DNA. These assays validated the cytomolecular results as follows: peak I on flow karyotypes contained chromosome groups 1, 4 and 6, peak II represented homoeologous group 5, while peak III consisted of groups 2, 3 and 7. The isolation of individual chromosomes of wild progenitors provides an attractive opportunity to investigate the structure and evolution of the polyploid genome and to deliver tools for wheat improvement.

Chromosome Preparation From Cultured Cells

Chromosome (cytogenetic) analysis is widely used for the detection of chromosome instability. When followed by G-banding and molecular techniques such as fluorescence in situ hybridization (FISH), this assay has the powerful ability to analyze individual cells for aberrations that involve gains or losses of portions of the genome and rearrangements involving one or more chromosomes. In humans, chromosome abnormalities occur in approximately 1 per 160 live births1,2, 60-80% of all miscarriages3,4, 10% of stillbirths2,5, 13% of individuals with congenital heart disease6, 3-6% of infertility cases2, and in many patients with developmental delay and birth defects7. Cytogenetic analysis of malignancy is routinely used by researchers and clinicians, as observations of clonal chromosomal abnormalities have been shown to have both diagnostic and prognostic significance8,9. Chromosome isolation is invaluable for gene therapy and stem cell research of organisms including nonhuman primates and rodents10-13. Chromosomes can be isolated from cells of live tissues, including blood lymphocytes, skin fibroblasts, amniocytes, placenta, bone marrow, and tumor specimens. Chromosomes are analyzed at the metaphase stage of mitosis, when they are most condensed and therefore more clearly visible. The first step of the chromosome isolation technique involves the disruption of the spindle fibers by incubation with Colcemid, to prevent the cells from proceeding to the subsequent anaphase stage. The cells are then treated with a hypotonic solution and preserved in their swollen state with Carnoy's fixative. The cells are then dropped on to slides and can then be utilized for a variety of procedures. G-banding involves trypsin treatment followed by staining with Giemsa to create characteristic light and dark bands. The same procedure to isolate chromosomes can be used for the preparation of cells for procedures such as fluorescence in situ hybridization (FISH), comparative genomic hybridization (CGH), and spectral karyotyping (SKY)14,15.

Chromosome preparation from cultured cells.

Chromosome (cytogenetic) analysis is widely used for the detection of chromosome instability. When followed by G-banding and molecular techniques such as fluorescence in situ hybridization (FISH), this assay has the powerful ability to analyze individual cells for aberrations that involve gains or losses of portions of the genome and rearrangements involving one or more chromosomes. In humans, chromosome abnormalities occur in approximately 1 per 160 live births(1,2), 60-80% of all miscarriages(3,4), 10% of stillbirths(2,5), 13% of individuals with congenital heart disease(6), 3-6% of infertility cases(2), and in many patients with developmental delay and birth defects(7). Cytogenetic analysis of malignancy is routinely used by researchers and clinicians, as observations of clonal chromosomal abnormalities have been shown to have both diagnostic and prognostic significance(8,9). Chromosome isolation is invaluable for gene therapy and stem cell research of organisms including nonhuman primates and rodents(10-13). Chromosomes can be isolated from cells of live tissues, including blood lymphocytes, skin fibroblasts, amniocytes, placenta, bone marrow, and tumor specimens. Chromosomes are analyzed at the metaphase stage of mitosis, when they are most condensed and therefore more clearly visible. The first step of the chromosome isolation technique involves the disruption of the spindle fibers by incubation with Colcemid, to prevent the cells from proceeding to the subsequent anaphase stage. The cells are then treated with a hypotonic solution and preserved in their swollen state with Carnoy's fixative. The cells are then dropped on to slides and can then be utilized for a variety of procedures. G-banding involves trypsin treatment followed by staining with Giemsa to create characteristic light and dark bands. The same procedure to isolate chromosomes can be used for the preparation of cells for procedures such as fluorescence in situ hybridization (FISH), comparative genomic hybridization (CGH), and spectral karyotyping (SKY)(14,15).

Induction of polyploidy by nuclear fusion mechanism upon decreased expression of the nuclear envelope protein LAP2β in the human osteosarcoma cell line U2OS

BackgroundPolyploidy has been recognized for many years as an important hallmark of cancer cells. Polyploid cells can arise through cell fusion, endoreplication and abortive cell cycle. The inner nuclear membrane protein LAP2β plays key roles in nuclear envelope breakdown and reassembly during mitosis, initiation of replication and transcriptional repression. Here we studied the function of LAP2β in the maintenance of cell ploidy state, a role which has not yet been assigned to this protein.ResultsBy knocking down the expression of LAP2β, using both viral and non-viral RNAi approaches in osteosarcoma derived U2OS cells, we detected enlarged nuclear size, nearly doubling of DNA content and chromosomal duplications, as analyzed by fluorescent in situ hybridization and spectral karyotyping methodologies. Spectral karyotyping analyses revealed that near-hexaploid karyotypes of LAP2β knocked down cells consisted of not only seven duplicated chromosomal markers, as could be anticipated by genome duplication mechanism, but also of four single chromosomal markers. Furthermore, spectral karyotyping analysis revealed that both of two near-triploid U2OS sub-clones contained the seven markers that were duplicated in LAP2β knocked down cells, whereas the four single chromosomal markers were detected only in one of them. Gene expression profiling of LAP2β knocked down cells revealed that up to a third of the genes exhibiting significant changes in their expression are involved in cancer progression.ConclusionsOur results suggest that nuclear fusion mechanism underlies the polyploidization induction upon LAP2β reduced expression. Our study implies on a novel role of LAP2β in the maintenance of cell ploidy status. LAP2β depleted U2OS cells can serve as a model to investigate polyploidy and aneuploidy formation by nuclear fusion mechanism and its involvement in cancerogenesis.

Effect of lenalidomide on inositide-dependent erythropoiesis and cell cycle in del(5q) myelodysplastic syndromes (mds)

Akt is implicated in erythropoiesis, through PI-PLCgamma1, but is also connected with cell cycle and protein synthesis, via activation of the mTOR pathway (1). Lenalidomide is currently used in the treatment of low-risk MDS patients showing a del(5q) karyotype, as it induces erythropoiesis in non-del(5q) cells and selectively inhibits cell proliferation in del(5q) cells (2). However, the exact molecular mechanisms underlying the effect of Lenalidomide in del(5q) and non-del(5q) MDS cells are still unclear, although Lenalidomide demonstrated an effect on targeting signalling molecules involved in apoptosis, proliferation and differentiation, i.e. Akt (3). Here we analyzed the effect of Lenalidomide on two cell lines: Namalwa CSN.70, bearing a del(5q) karyotype, and U937, with a normal 5q chromosome. In particular, we analyzed several molecules implicated in inositide signalling, such as Akt, mTOR and PI-PLCgamma1, as well as Cyclins and Globin genes, in order to assess the effect of Lenalidomide on cell cycle and erythropoiesis. Moreover, we quantified the gene expression profile of 6 patients diagnosed with del(5q) Low-Risk MDS (IPSS: Low or Int-1) who were given Lenalidomide. In our case series, as well as in cell lines, erythropoiesis activation was associated with a response to Lenalidomide, with an induction of Akt/PLCgamma1 and an increase of Beta-Globin. Moreover, only in non-del(5q) cells, a normal proliferation was allowed, given that del(5q) cells showed a cell cycle arrest and a slight inhibition of the Akt/mTOR pathway, this latter being confirmed also by co-localization experiments performed on primary cells from MDS patients. Therefore, our data support the hypothesis of a specific activation of both inositide-dependent proliferation and erythroid differentiation pathways in response to Lenalidomide treatment in non-del(5q) cells, whereas in the del(5q) cell clone there is a cell cycle arrest and a slower erythroid differentiation. Taken together, these results hint to a specific activation of inositide-dependent signalling pathways during Lenalidomide administration and possibly pave the way to a larger investigation aiming to assess the role of these pathways during the therapy.

Diagnostic yield and cost-utility analysis of genetic investigations for assessing children with autism in an australian metropolitan child development service

Aim This study aimed to determine the utilisation, yield and cost of screening genetic investigations in a sample of children diagnosed with pervasive developmental disorders (PDD; DSM-IV). The purpose was to inform development of strategies for genetic investigation of children with these disorders. Methods 699 patients with PDD were identified, and genetic investigations evaluated. A sub-group of 219 patients underwent clinical chart audit, and the resulting clinical characteristics evaluated in the context of investigation use and findings. Results 24.8%, 15.9% and 25.1% of the clinic population had undergone investigation with chromosomal microarray (CMA), karyotype (KT) and fragile X testing, respectively. CMA is associated with a cost reduction of $3437 less per diagnosis compared with KT (p = 0.026). Several characteristics were associated with increased odds of a significant CMA finding, most significant of which was comorbid epilepsy (odds ratio 10.4; p Discussion CMA appears to increase the diagnostic yield of screening genetic investigations in PDD. This finding is consistent with what has been seen in other populations internationally. CMA reduces cost per incremental diagnosis with comparison to KT, amongst children with PDD. Epilepsy was found to be a significant predictor of a pathogenic finding on chromosomal microarray.

Characterization of Four Specimens of&lt;i&gt; Allium sativum&lt;/i&gt; L. by Differential Karyotype and RAPD Analysis

Characterization of Four Specimens of&lt;i&gt; Allium sativum&lt;/i&gt; L. by Differential Karyotype and RAPD Analysis

18p deletion syndrome: foetal phenotype and cytogenetic characterization

Monosomy for the short arm of chromosome 18 is one of the most common autosomal deletion syndromes, with an estimated incidence of about 1:50000 live-born. The phenotype of this chromosome imbalance is greatly variable, often not evident at birth. In this communication we describe a case prenatally diagnosed of de novo 18p deletion.Amniocentesis was performed at 17 weeks of gestational age on a 39-year-old gravid. The family history was unremarkable and the pregnancy was uncomplicated. At the time of the amniocentesis the amniotic fluid alphafetoprotein level was slightly lower than normal. The pregnancy was terminated at 18 weeks, after the cytogenetic response. At the time the ultrasonographic examination showed reduced biparietal diameter and head circumference, normal limbs, normal thoracic, abdominal and pelvic organs. The external examination of the aborted foetus showed: weight 380 gr; ipsicephaly and nuchal oedema, no facial dysmorphic signs, normal ears, limbs of normal length, overlapping toes in the right foot. External genitalia were normal. At the autopsic examination the right lung showed four lobes; all the other organs of thorax, abdomen and pelvis were normal.Cytogenetic analysis of cultured amniocytes showed a de novo 18p deletion. FISH studies allowed us to define the foetal karyotype as: 46,XX, del(18)(p10pter).ish del(18)(tel18p-, dim D18Z1). The phenotypic spectrum of 18p- syndrome may include: round face, ptosis, flat and broad nasal bridge, wide mouth with short upper lip, small mandibles, irregular teeth and dental caries, large, protruding ears, often low and posteriorly rotated, short neck, pectus excavatum, kyphoscoliosis, hands and feet anomalies. The main malformations are holoprosencephaly (HPE), cardiac defects and genital anomalies. Growth retardation, psychomotor delay and mental insufficiency, often mild, are constant symptoms of the syndrome. The foetal phenotype is inconsistent, but the increased nuchal translucency (INT) seems to be a sonographic sign suggestive of this chromosomal abnormality. This further stress the importance of accurate morphological ultrasonographies, mainly in the second trimester, in order to prevent rare and oligosymptomatic chromosomal syndromes.

Karyotypic characterisation and C-banding of representatives from Geotrupinae and Dynamopinae (Coleoptera: Scarabaeidae)

Karyotypic characterisation and C-banding of representatives from Geotrupinae and Dynamopinae (Coleoptera: Scarabaeidae)

Tumor necrosis factor-α and NF-κB play a role in macrophage-like THP-1 cells promoting coal tar pitch extract-induced tumorigenic transformation of human bronchial epithelial cells

To characterize the role of tumor necrosis factor-α (TNF-α) and NF-κB play a role in macrophage-like THP-1 cells promoting coal tar pitch extract (CTPE)-induced tumorigenic transformation of human bronchial epithelial cells (BEAS-2B). From passage 10, CTPE-induced BEAS-2B cells cocultured with THP-1 cells were treated with NF-κB inhibitor-Pyrrolidine dithiocarbamate (PDTC) every 3 passages and TNF-α antibody every passage. Alterations of cell cycle, karyotype and colony formation in soft agar of BEAS-2B cells at passages 20, indicative of tumorigenicity, were determined, respectively. In addition, mRNA and protein levels of TNF receptor associated factor2 (TRAF2) and Cyclin D1 in BEAS-2B cells were measured with Real Time-PCR and Western blot, respectively. The percentages of S-phase BEAS-2B cells at passage 20 in PDTC group and TNF-α antibody group were (33.97±2.16)% and (34.29±2.04)% respectively, which were less than that in Co-culture+CTPE group of 20th passage [(44.46±0.83)%], P < 0.05; The number of cells with aneuploidy in 100 cells in 20th passage PDTC group and TNF-α antibody group were 40 and 37, and there were significantly different when comparing to that of 20th passage Co-culture+CTPE group (75); The number of colony formation and the rate of colony formation of BEAS-2B cells in soft agar at passage 20 in PDTC group were (15.17±2.48) and (1.51‰±0.25‰), (13.33±2.58)and (1.33‰±0.26‰) in TNF-α antibody group, which were less that those in 20th passage Co-culture+CTPE group [(172.33±12.09) and (17.23‰±1.20‰)], P < 0.05; at the same time, the mRNA and protein levels of TRAF2 and Cyclin D1 in BEAS-2B cells were decreased after PDTC and TNF-α antibody treatment. TNF-α and NF-κB could play an important role in THP-1 cells promoting coal tar pitch extract-induced tumorigenic transformation of BEAS-2B cells by influencing the expression of TRAF2 and Cyclin D1.

Establishment and characterization of a highly tumorigenic African American prostate cancer cell line, E006AA-hT.

Genuine racial differences in prostate cancer (PCa) biology have been considered among the potential reasons to explain PCa disparities. There is no animal model to represent all aspects of human PCa and, more specifically, to be used for PCa disparity research. The lack of a spontaneously transformed in vitro cell-based model system has been a significant impediment to investigating and understanding potential molecular mechanisms, and the hormonal, genetic, and epigenetic factors underlying the biological and clinical aggressiveness of PCa in African American (AA) men.In this study, we established and characterized the E006AA-hT cell line as a highly tumorigenic subline of the previously characterized primary AA-PCa cell line, E006AA. Extensive characterization of the E006AA-hT cell line was accomplished using cytodifferentiation and prostate-specific markers, spectral karyotyping, cell line authentication assays, cell proliferation and migration assays, and in vitro tumorigenesis assays.Spectral karyotyping of E006AA-hT showed a hypertriploid chromosome complement and shared cytogenetic changes similar to its parental cells such as diploid X, absence of Y-chromosomes, numerical gains in chromosomes 5,6,8,10,17,20,21, and marker chromosomes of unknown origin. In addition, E006AA-hT also presented numerous clonal and structural aberrations such as insertion, deletion, duplication, and translocations in chromosomes 1-5, 8, 9, 11, 13, 14, 17, and 18. The E006AA-hT cell line was shown to be highly tumorigenic and produced tumors at an accelerated growth rate in both athymic nude and triple-deficient SCID mice.Silencing the mutated androgen receptor (AR-599 Ser>Gly) did not affect proliferation (loss-of-function), but decreased migration (gain-of-function) in E006AA-hT and its parental cell type. These data support that AR-point mutations may lead simultaneously to different “loss-of-function” and “gain-of-function” phenotypes in PCa cells. E006AA-Par and its subline as the only available spontaneously transformed low- and highly-tumorigenic primary AA-PCa cell lines could be used for basic and translational research aimed in supporting prostate cancer disparity research.