Search

The karyotypes of Tulipa sinkiangensis and T. schrenkii were analyzed based on chromosome size and the physical mapping of 5S and 45S ribosomal DNAs (rDNAs) using fluorescence in situ hybridization. The genomes from both species were consistent with the uniform karyotype formula, i.e., having four pairs of submedian and eight pairs of subterminal chromosomes (2 n =2 x =24=8sm+16st). Remarkably, the number and location of 5S rDNA loci were quite different between the two species. A single pair of 5S rDNA loci was detected on both chromosomes 5 in T. sinkiangensis , but the 5S rDNA loci were detected on almost all chromosomes in T. schrenkii , except for chromosomes 4 and 6. For both species, the 45S rDNA loci were localized to telomeric regions of the chromosome—on either the short or long arm. Interestingly, the 5S rDNA and 45S rDNA loci positions and numbers varied among different genotypes of T. schrenkii . We conclude that the two species are close relatives and that a series of genome modifications occurred during the diversification of these two species. [ABSTRACT FROM AUTHOR]

Saccharina japonica is an economically and ecologically important brown alga. The metaphase chromosomes of this kelp were small in size and similar in length, which had hampered its karyotype analysis. In recent decade, a combination of DNA-intercalating dye 4′, 6-Diamidino-2-phenylindole (DAPI) and fluorescence in situ hybridization (FISH) technique had promoted the karyotype analysis of S. japonica progressively. Till now, a set of molecular markers including 18S and 5S rDNAs, sex-linked molecular markers, telomeric repeated sequences, as well as SSRs had been used in karyotyping of this kelp by using FISH, dual-color FISH, and fiber-FISH. In addition, immunocytochemistry with kinetochore protein were attempted for its more accurate chromosome analysis. As the results, 31 and 31 pairs of chromosomes were observed in haploid and diploid cell respectively, among which 11 or 11 pairs of chromosomes with telomeric probe signals at both ends were confirmed. Notably, the putative U/V sex chromosomes had also been discerned. On the basis of above results, it was suggested that centromeric DNA markers should be developed for more precise karyotype analysis of this kelp. In addition, the developed molecular markers as well as the established method systems in these studies would be helpful to the karyotype analysis of other macroalgae. • The lack of distinguishing features among chromosomes had hampered the karyotyping of Saccharina japonica. • Molecular markers had been developed for chromosome identification. • FISH, dual-color FISH, and fiber-FISH had been performed in karyotype analysis of this kelp. • 31 and 62 chromosomes had been observed in haploid and diploid cells of S. japonica , respectively. • 11 pairs of chromosomes had been confirmed, and the putative U and V chromosomes had been discerned. [ABSTRACT FROM AUTHOR]

Purpose: Evaluation of the relationships existing among 3 histologic types of urothelial tumors, chromosomal instability, and telomere length. Patients and methods: We examined 37 consecutive cases of papillary urothelial neoplasm, from which 26 (70.3%) were suitable for karyotype analysis, comprising 7 papillary urothelial neoplasms of low malignant potential (PUNLMPs), 10 low-grade papillary urothelial carcinomas (PUCs), and 9 high-grade PUCs. We performed karyotype and anaphase bridge analyses, and measured telomere lengths by quantitative fluorescence in situ hybridization. Results: PUNLMPs were always diploid and had anaphase bridges. Low-grade PUCs showed diploidy (n = 2), hypoploidy (n = 4) and polyploidy (n = 4), and high-grade PUCs showed diploidy (n = 1) and polyploidy (n = 8); both had anaphase bridges. The incidence of anaphase bridges did not differ significantly between PUNLMPs and high-grade PUCs (P = 0.105). The telomere lengths of PUNLMP, low-grade PUC, and high-grade PUC, expressed as mean telomere fluorescence units (TFU)±SD, were 7906±3197, 4893±1567, and 3299±1406, respectively. The differences among the 3 groups were significant. However, 42.9% of the PUNLMPs had shorter telomeres than the mean value for low-grade PUCs, and 30.0% of the low-grade PUCs had shorter telomeres than those for high-grade PUCs. There was an inverse correlation between telomere length and the incidence of anaphase bridges. Conclusions: PUNLMP appears to progress to low-grade PUC and high-grade PUC in association with telomere shortening and chromosomal instability. Our data suggest that critically shortened telomeres cause chromosomal instability during progression of papillary urothelial neoplasms. [ABSTRACT FROM AUTHOR]

Abstract: Here we attempted to clarify telomere metabolism in parental cells and their derived clonal human induced pluripotent stem cells (iPSCs) at different passages using quantitative fluorescence in situ hybridization (Q-FISH). Our methodology involved estimation of the individual telomere lengths of chromosomal arms in individual cells within each clone in relation to telomere fluorescence units (TFUs) determined by Q-FISH. TFUs were very variable within the same metaphase spread and within the same cell. TFUs of the established iPSCs derived from human amnion (hAM933 iPSCs), expressed as mean values of the median TFUs of 20 karyotypes, were significantly longer than those of the parental cells, although the telomere extension rates varied quite significantly among the clones. Twenty metaphase spreads from hAM933 iPSCs demonstrated no chromosomal instability. The iPSCs established from fetal lung fibroblasts (MRC-5) did not exhibit telomere shortening and chromosomal instability as the number of passages increased. However, the telomeres of other iPSCs derived from MRC-5 became shorter as the number of passages increased, and one (5%) of 20 metaphase spreads showed chromosomal abnormalities including X trisomy at an early stage and all 20 showed abnormalities including X and 12 trisomies at the late stage. [Copyright &y& Elsevier]

Objective: To compare the roles of fluorescence in situ hybridization (FISH) and karyotype analysis on chorionic villus in spontaneous abortion. Methods: A total of 201 cases were included in this study and were randomly divided into 2 groups by the same gestational age. The villi from 100 cases (group A) were cultured and used for karyotype analysis, while the uncultured villi from 101 women (group B) were used for FISH. A case was randomly selected from pregnant patients of 6–11 weeks at each gestational week in each group, 6 cases from each group and a total of 12 cases were analyzed by FISH and karyotype at the same time. Results: The successful karyotype analysis rate was 66.0%, and the abnormality karyotype rate was 30.3%; FISH success analysis rate was 100%, and the abnormality rate was 46.5%; there were significant differences between FISH and karyotype analysis (P=0.036). Conclusion: There were obvious differences between the two techniques. FISH was more successfully analyzed, and was used to more precisely determine fetal chromosome number. It is an effective way to determine abnormal chromosome by integrating FISH and karyotype analysis in spontaneous abortion. [Copyright &y& Elsevier]

Abstract: We describe the prenatal diagnosis and fetal phenotype of partial trisomy 12 (p12-pter) transmitted from a maternal reciprocal translocation 6;12. Genetic analysis was conducted on umbilical cord blood for a fetus accompanied with tricuspid regurgitation and orbital hypertelorism from a 27-year-old gravida 4, para 1 after sonography at gestation 35weeks. The karyotype was unusual, with 46, XY, der (6), t (6;12) (p24; p12) mat. The pregnancy was terminated at 37 gestational weeks. The aborted fetus displayed dysmorphic features of a round flat face with prominent cheeks and high forehead, hypertelorism, short nose, broad and depressed nasal bridge, anteverted nares, deformed philtrum, open mouth, thin upper vermilion and broad everted lower lip, low-set ears and aural atresia, broad hands with simian creases, and a short neck. Fetal anatomy showed right artery catheter vagus, congenital cataract, no turbinate and external auditory canals. Through karyotype–phenotype analysis of this patient and a review of other reported cases, we believe this is a first report that expands the database of partial trisomy 12p, and is beneficial for future clinical genetic counseling. This study supports that phenotypic variability depends on the type and extent of the associated partial monosomy. [Copyright &y& Elsevier]

Abstract: We describe the prenatal diagnosis and fetal phenotype of partial trisomy 12 (p12-pter) transmitted from a maternal reciprocal translocation 6;12. Genetic analysis of umbilical cord blood of a 27-year-old woman, gravida 4, para 1 at 35 weeks'' gestation due to a tricuspid regurgitation and orbital hypertelorism by sonography revealed an unusual karyotype of 46, XY, der (6) t (6;12) (p24;p12) mat. The pregnancy was terminated at 37 gestational weeks. The proband postnatally displayed by dysmorphic features of a round flat face with prominent cheeks and high forehead, hypertelorism, a short nose, a broad and depressed nasal bridge, anteverted nares, a deformed philtrum, an open mouth, thin upper vermilion and broad everted lower lip, low-set ears and aural atresia, broad hands with simian creases, and a short neck. By anatomy, the fetal was found to have right artery catheter vagus, congenital cataract, no turbinate and external auditory canal. Through the karoytpye-phynotpye analysis on the present patient and a review of other reported cases, we believed that the case was the first report, which expanded the database of partial trisomy 12p, and was of benefit for future clinical genetic counseling. At the same time, this study supported the viewpoint that phenotypic variability depends on the type and extent of the associated partial monosomy. [Copyright &y& Elsevier]

Background: Neonates with a single umbilical artery (SUA) are considered at increased risk for chromosomal and structural abnormalities, and an increased adverse perinatal outcome. Objective: The specific aims of our study were to evaluate (1) the association of asymptomatic infants with isolated SUA and perinatal outcomes and (2) whether asymptomatic neonates with isolated SUA at birth need full investigation. Methods: The inclusion criteria for the study were full-term neonates with isolated SUA delivered from January 1996 to December 2006. For a control group, we used the next consecutive two newborns delivered after the SUA case in the same maternity ward with matched gestational age and without phenotypic features suspicious for aneuploidy delivered after each SUA group subject. All prenatal, peripartum and delivery records were reviewed for maternal demographics, associated anomalies, karyotypic analysis, pregnancy complications and perinatal outcomes. All SUA cases had undergone sonogram for renal anomalies. Results: We enrolled 14 and 28 cases into the SUA and control groups respectively. There was all normal karyotyping for the 14 cases. The placental weight in SUA was significantly lighter compared to that in the control group (597.1 ± 175.4 vs. 709.3 ± 95.2 g, p = 0.010). All renal sonographic screens and karyotyping in the SUA group were normal. The incidence of small for gestational age (SGA) in SUA group was higher compared to control group (SGA, 5/14, 35.7%vs. 1/28, 3.6%, p= 0.011) and less body length (48.7 ± 5.0 vs. 50.8 ± 1.8 cm, p = 0.028). Conclusion: SUA is a relatively rare finding. When a SUA is identified, the routine check of karyotyping and kidney sonography for possible chromosome and associated renal anomalies may be unnecessary. According to lighter placental weight probably causing the higher incidence of small for gestational age (SGA), pregnancies with isolated SUA should be carefully monitored for evidence of fetal growth restriction. [Copyright &y& Elsevier]

Abstract: The small ruminant lentiviruses, namely caprine arthritis encephalitis virus (CAEV) and Maedi Visna virus (MVV) are grown currently in secondary synovial membrane cells. Primary and secondary cell cultures are sometimes difficult to obtain and support a low number of passages and, therefore, permissive cell lines are needed. A transformed cell line was obtained by transfection of ovine synovial membrane secondary cell culture with a plasmid containing the SV40 large T antigen gene. The transformed cell culture described in this paper showed a higher growth rate and a more homogenous population of fibroblast-like cells when compared to the original ovine synovial membrane secondary cell cultures. Karyotype analysis has indicated the induction of many random chromosome changes, leading to a decrease in chromosome number. The SV40 DNA was detected in the nucleus and in the cytoplasm of transformed cells. The putative expression of large T antigen was presumed by the detection of the corresponding mRNA by PCR. Finally, the transformed ovine synovial membrane cells were shown to be permissive to small ruminant lentiviruses, and these are suggested as a cell line for in vitro isolation and propagation of these viruses. [Copyright &y& Elsevier]

Summary: This article analyzes phenotype and genotype alterations of the lung in association with lung cancer. The frequency of phenotype preneoplastic lesions (atypical adenomatoid hyperplasia (AAH) and squamous cell dysplasia (SCD)) was analyzed at distinct distances from the tumor boundary in 150 lung carcinomas. AAH was noted in 19/150 (13%) cases and more frequently seen in adeno carcinomas, squamous cell dysplasia was noted in 46/150 (31%) cases and more frequently seen in squamous cell carcinomas. The degree of cellular atypia decreased with increasing distance from tumor boundary in both AAH and SCD. At similar distances, genotype (chromosome) alterations of surrounding bronchial mucosa were studied in additional 55 primary and secondary lung tumors by karyotype analysis. Numerical chromosome aberrations occur frequently in primary lung carcinomas and adjacent bronchial mucosa, and affect at average 4.5/10 metaphases in primary lung cancer and 2/10 metaphases in metastases. Most abnormal metaphases were induced by chromosome losses, only few by additional copies, i.e. trisomy, etc. Losses of y chromosome were seen in both malignancy and adjacent bronchial mucosa, and interpreted as “tumor related”, losses of chromosome 21 in adjacent bronchial mucosa were non-tumor related in adenocarcinoma and metastases, losses of chromosome 19 in adjacent bronchial mucosa occurred independently in squamous cell and large cell carcinomas. The data suggest the hypothesis that preneoplastic lesions in the lung might be partly induced by the tumor itself. [Copyright &y& Elsevier]

Dojo loach (Misgurnus anguillicaudatus) (DD, 2n = 50) and large-scale loach (Paramisgurnus dabryanus) (PP, 2n = 48) are freshwater fish species widely distributed across East Asia. Hybrids of these two loach species have disrupted gonadal development, but few studies have reported the actual fertility and chromosome number in gametes of reciprocal hybrid loaches (DD × PP and PP × DD, assigned as DP and PD, 2n = 49). In this study, selfing and backcrossing experiments between M. anguilicaudatus and P. dabryanus were conducted. The fertilization, hatching, deformity, and survival rates were examined along with the embryonic development and growth performance to analyze the actual fertility of the hybrid loach. The karyotypes of viable backcross offspring were studied to indirectly assess the chromosome number in eggs of DP and PD hybrids. The failure in fertilization among crosses using male hybrid proved that the male hybrid was sterile. The moderate fertilization rates (11.49%‐–49.16%), high hatching rates (82.78%–86.31%), high survival rates (78.67%–81.33%), and the normal embryonic development among crosses using female hybrid revealed that the female hybrid was fertile, albeit with relatively high deformity rates (12.68%–27.45%). The karyotype analysis of viable backcross offspring showed 49 chromosomes for DP × DD (assigned as DPD) and PD × DD (assigned as PDD) hybrids, and 48 chromosomes for DP × PP (assigned as DPP) and PD × PP (assigned as PDP) hybrids. This indicated that fertile gametes produced by female hybrid loach contained 24 chromosomes. This study provides valuable information for loach hybrid breeding and in-depth research on the precise mechanism of meiosis in these hybrid loach. • The fertility of reciprocal Misgurnus anguillicaudatus and Paramisgurnus dabryanus hybrids was explored. • The female hybrid loach was fertile. • The male hybrid was sterile. • The fertile gametes produced by female hybrids contained 24 chromosomes. [ABSTRACT FROM AUTHOR]

• An adaptive receptive field multi-scale network (ARMS Net) based on UNet architecture is proposed to segment chromosomes that curled and overlapped together randomly in different images. • The number of pooling operations is optimized to balance the requirements of deep semantic information extraction and high precision segmentation. • The adaptive multi-scale feature extraction module is designed to replace the standard convolution at the bottom of UNet, such that the receptive fields can adaptively match the size of feature map. • An adaptive smooth weighted cross entropy loss function is defined to resolve sample category unbalance issue. Karyotype analysis has become the crux for diagnosis of genetic diseases, which requires automatic precise identification of quantitative and structural abnormalities. Thus, segmentation of chromosomes from microscope captured photos by image processing technique can help promote the automatic identification of chromosomes. However, chromosomes are often curled and overlapped together randomly in different images, which makes overlapping chromosome segmentation the hot topic in karyotype analysis. Herein an Adaptive Receptive field Multi-Scale network (ARMS Net) based on UNet architecture is proposed. The number of pooling operations is optimized to balance the requirements of deep semantic information extraction and high precision segmentation. The adaptive multi-scale feature extraction module is designed to replace the standard convolution at the bottom of UNet, such that the receptive fields can adaptively match the size of feature map. Besides, an adaptive smooth weighted cross entropy loss function is defined to resolve category imbalance issue. Experimental results show that the Intersection of Union (IoU) score of ARMS Net segmented overlapping area is 99.45%, which is 3.2% higher than that achieved by UNet (96.38%), and 10.1% higher than that achieved by CE-Net (90.35%). In a word, ARMS Net is expected to be used as the backbone network for chromosome instance segmentation in its end-to-end identification. [ABSTRACT FROM AUTHOR]

Abstract: Humans with mosaic karyotypes may present with milder forms of birth defects than one would see in non-mosaic individuals. Mosaicism may also affect tissues, resulting in different parts of an organ manifesting varied phenotypes. We present the case of a child born with mosaic trisomy 9 and multiple congenital anomalies. Her trachea displays segmental abnormalities that suggest tissue mosaicism. We describe the endoscopic and cytogenetic findings in this child and propose a possible genetic mechanism to account for the unusual malformations. [Copyright &y& Elsevier]

Objective To demonstrate a technique for separation of miscarriage tissue from maternal decidua to reduce maternal cell contamination for chromosome analysis. Design Retrospective. Setting University-based infertility center. Patient(s) Not applicable. Intervention(s) Retrospective collection of de-identified images and video from manual vacuum aspiration (MVA) performed after first-trimester pregnancy losses. This project was exempt from institutional review board approval as no patient-identifying data were used. Main Outcome Measure(s) Reduction of maternal cell contamination and improvement of the accuracy of chromosome analysis. Result(s) Video demonstration of separation of miscarriage tissue from maternal decidua after MVA. Conclusion(s) Chromosome analysis of a miscarriage is performed to assess the etiology of miscarriage and recurrent pregnancy loss. However, maternal cell contamination can limit the accuracy. This video demonstrates a technique for separating miscarriage tissue from maternal decidua after MVA to reduce maternal cell contamination prior to sending tissue for analysis. The same technique has been used in our clinic with first-trimester dilation and curettage using mechanical suction. [ABSTRACT FROM AUTHOR]