OBJECTIVE: To describe in vitro development of human embryos derived from an individual with a homozygous pathogenic variant in NLRP7 (19q13.42) and recurrent hydatidiform mole (HM), an autosomal recessive condition thought to occur secondary to an oocyte defect. METHODS: A patient with five consecutive HM pregnancies was genomically evaluated via next generation sequencing followed by controlled ovarian hyperstimulation, in vitro fertilization (IVF) with intracytoplasmic sperm injection, embryo culture, and preimplantation genetic screening. Findings in NLRP7 were recorded and embryo culture and biopsy data were tabulated as a function of parental origin for any identified ploidy error. RESULTS: The patient was found to have a pathogenic variant in NLRP7 (c.2810+2T>G) in a homozygous state. Fifteen oocytes were retrieved and 10 embryos were available after fertilization via intracytoplasmic sperm injection. Developmental arrest was noted for all 10 embryos after 144 hours in culture, thus no transfer was possible. These non-viable embryos were evaluated by karyomapping and all were diploid biparental; two were euploid and eight had various aneuploidies all of maternal origin. CONCLUSION: This is the first report of early human embryo development from a patient with any NLRP7 mutation. The pathogenic variant identified here resulted in global developmental arrest at or before blastocyst stage. Standard IVF should therefore be discouraged for such patients, who instead need to consider oocyte (or embryo) donation with IVF as preferred clinical methods to treat infertility.
Abortion, Habitual ; Aneuploidy ; Biopsy ; Blastocyst ; Diploidy ; Embryonic Development* ; Embryonic Structures* ; Female ; Fertilization ; Fertilization in Vitro* ; Genetic Testing ; Gestational Trophoblastic Disease* ; Humans ; Hydatidiform Mole ; In Vitro Techniques* ; Infertility ; Oocytes ; Parents ; Ploidies ; Pregnancy ; Sperm Injections, Intracytoplasmic

BACKGROUND: The accurate identification of cytogenetic abnormalities in multiple myeloma (MM) has become more important over recent years for the development of new diagnostic and prognostic markers. In this study, we retrospectively analyzed the cytogenetic aberrations in MM cases as an initial assessment in a single institute. METHODS: We reviewed the cytogenetic results from 222 patients who were newly diagnosed with MM between January 2000 and December 2015. Chromosomal analysis was performed on cultured bone marrow samples by standard G-banding technique. At least 20 metaphase cells were analyzed for karyotyping. RESULTS: Clonal chromosome abnormalities were detected in 45.0% (100/222) of the patients. Among these results, 80 cases (80.0%) had both numerical and structural chromosome abnormalities. Overall hyperdiploidy with structural cytogenetic aberrations was the most common finding (44.0%), followed by hypodiploidy with structural aberrations (28.0%). Amplification of the long arm of chromosome 1 and -13/del(13q) were the most frequent recurrent abnormalities, and were detected in 50 patients (50.0%) and 40 patients (40.0%) with clonal abnormalities, respectively. The most common abnormality involving 14q32 was t(11;14)(q13;q32), which was observed in 19 cases. CONCLUSION: These findings demonstrate that myeloma cells exhibit complex aberrations regardless of ploidy, even from a single center in Korea. Conventional cytogenetic analysis should be included in the initial diagnostic work-up for patients suspected of having MM.
Arm ; Bone Marrow ; Chromosome Aberrations ; Chromosomes, Human, Pair 1 ; Cytogenetic Analysis ; Cytogenetics* ; Humans ; Karyotyping ; Korea* ; Metaphase ; Multiple Myeloma* ; Ploidies ; Retrospective Studies*

OBJECTIVETo explore the clinical values of detecting immunophenotype and analyzing DNA ploid by flow cytometry for patients with non-Hogkin's lymphoma (NHL).

METHODSEighty NHL patients admitted in our hospital from August 2007 years to March 2015 Years were included in the observation group, 20 patients with reactive lymphoid hyperplasias were selectod as control group. The immunophenotypes were detected and the DNA ploid was analyzed by flow cytometry.

RESULTSThe detected rate of DNA aneuploidy, DAN index (DI) and SPF in observation group were higher than those in control group, there was signifificant difference (P < 0.05). The SPF and DI in patients with NHL-I, NHL-II had no statistical difference as compared with that in control group (P > 0.05); but the SPF and DI in pateints with NHL-III and patients with NHL-IV showed statistical significance as compared with that in control group (P < 0.05). The SPF and DI in patients with low malignancy group and middle malignancy group showed statistical significance as compared with control group (P < 0.05). The SPF and DI in middle malignancy group had statistical significance as compared with that in low malignancy group (P < 0.05).

CONCLUSIONthe immunophenotype detection and DNA ploid analysis by flow cytometry can reflect the tumor proliferation and deterioration of patients with Non-Hogkin's lymphoma, predicting the prognosis.

Aneuploidy ; Case-Control Studies ; DNA ; Flow Cytometry ; Humans ; Immunophenotyping ; Lymphoma, Non-Hodgkin ; classification ; genetics ; Ploidies ; Prognosis

INTRODUCTION: Hydatidiform mole (HM) is an abnormal gestation characterized by significant hydropic enlargement, trophoblastic proliferation and atypia involving part or all of the chorionic villi. The diagnosis and classification of hydatidiform moles is subject to great inter-observer variability due to significant morphologic overlaps. This study aims to evaluate the utility of p57KIP2 immunohistochemistry and ploidy by Her-2 FISH in refining the diagnosis of molar tissues.

METHOD: 113 and 78 molar cases were retrieved from the archives of the Histopathology Section of the Philippine General Hospital and Pathology Department of Seoul National University Hospital, respectively. TMA sections were submitted for immunohistochemical analysis for p57KIP2. Ploidy was determined by fluorescence in situ hybridization using Her-2 probe. An interrater reliability analysis was done using the Kappa statistics with 95% confidence interval.

RESULTS: All 68 (100%) cases diagnosed as CH were negative for p57KIP2 staining and are diploid. Among the 54 cases of PH, only 1 (2%) is positive for p57KIP2 and is diploid. The interrater reliability between p57KIP2 and Her-2 FISH ploidy results is 0.66 (p <.0.001), 95% CI (0.02, 1.00) which is considered "fair to good." The kappa value between review diagnosis and p57KIP2 is 0.024 while the kappa between review diagnosis and Her-2 FISH ploidy is 0.050 both signifying poor agreement beyond chance.

CONCLUSION: Morphologic assessment alone may not be sufficient in problematic cases. p57KIP2 in conjunction with by Her-2 FISH are good adjuncts in the diagnosis and classification of hydatidiform mole.

Human ; Male ; Female ; Pregnancy ; Chorionic Villi ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Diploidy ; Reproducibility Of Results ; Hydatidiform Mole ; Trophoblasts ; Ploidies ; Molar

OBJECTIVE: It has previously been suggested that embryos developing from intracytoplasmic sperm-injected (ICSI) zygotes with three pronuclei (3PN) are endowed with a mechanism for self-correction of triploidy to diploidy. 3PN are also observed in zygotes after conventional in vitro fertilization (IVF). The parental origin, however, differs between the two fertilization methods. Whereas the vast majority of 3PN IVF zygotes are of dispermic origin and thus more likely to have two centrioles, the 3PN ICSI zygotes are digynic in origin and therefore, more likely to have one centriole. In the present study, we examine whether the parental origin of 3PN embryos correlates with the karyotype. METHODS: The karyotype of each nucleus was estimated using four sequential fluorescence in situ hybridizations-each with two probes-resulting in quantitative information of 8 different chromosomes. The karyotypes were then compared and correlated to the parental origin. RESULTS: 3PN ICSI embryos displayed a significantly larger and more coordinated reduction from the assumed initial 3 sets of chromosomes than 3PN IVF embryos. CONCLUSION: The differences in the parental origin-and hence the number of centrioles-between the 3PN IVF and the 3PN ICSI zygotes are likely to be the cause of the differences in karyotypes.
Centrioles ; Diploidy ; Embryonic Structures* ; Fertilization ; Fertilization in Vitro ; Fluorescence ; Humans ; Karyotype* ; Parents* ; Ploidies ; Sperm Injections, Intracytoplasmic ; Triploidy ; Zygote*

This study was aimed to investigate the genetic characteristics of human acute lymphoblastic leukemia cell line Molt-4, and evaluate its application in measuring telomere length by Flow-FISH. Molt-4 cell line was cultured in suspension and subcultured regularly. Eight different passages of Molt-4 cells in exponential stage were selected.The growth curves were drawn by cell counting method, meanwhile calculating the population doubling times of cells,DNA ploidies were determined by flow cytometry,karyotypes were analyzed by G-banding and telomere lengths were measured by Southern blot. The results showed that the population doubling time of Molt-4 cell line was (1.315 ± 0.062) d, DNA ploidy index was (2.085 ± 0.0093) , and the telomere length was (32.05 ± 5.27) kb. There were no significant difference among different passages (P = 0.931,0.888 and 0.935 separately). The karyotypes showed that the chromosome numbers of Molt-4 cell line were from 91 to 99 in different metaphases, and the majority of them were hypertetraploid, and stable and recurrent structural abnormalities of chromosomes could be kept. It is concluded that the stable genetic characteristics and the longer telomere length of Molt-4 cell line makes it be a feasible control cells in measurement of telomere length by Flow-FISH.
Cell Line, Tumor ; DNA, Neoplasm ; Flow Cytometry ; Humans ; Karyotyping ; Ploidies ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Telomere ; genetics

In order to broaden Chinese cabbage gene pool, we conducted interspecific somatic hybridization between Chinese cabbage (Brassica campestris, 2n=20, AA) and Cabbage (B. oleracea, 2n=18, CC). Protoplasts were isolated from 10-day-old cotyledons and hypocotyls of young seedlings, and fused by 40% polyethylene glycol (PEG). Fused cells were cultured in modified K8p liquid medium supplemented with some plant growth regulators. Fusion products were characterized by their morphological, cytological and molecular biological traits. The results showed that, a total of 35 regenerated green plants were obtained from 320 calli, the plant regeneration frequency was 10.94%, and eleven of which were survived in greenhouse. All regenerants were true hybrids as confirmed by randomly amplified polymorphic DNA (RAPD) and genomic in situ hybridization (GISH) analysis. Ploidy levels of hybrid plants were determined by chromosome counting and flow cytometry. The sum of the chromosome number (2n = 38) from the two fusion patents were found in 36.4% of regeneratns; another 36.4% had chromosomes range to 58-60; 27.2% had more chromosomes ranges to 70-76. All regenerated plants produced normal flowers. We investigated the pollen fertility and seed set after self-pollination and backcrossing with the parental species. For hybrids with chromosomes more than 38 it was possible to obtain some seeds when they after self-pollination. Within the group of hybrids with 38 chromosomes, seed set were very variable, only 0.11 seeds per pod by self-pollination, 0.23-0.76 by open-pollination, 0.02-0.04 by backcrossing with Chinese cabbage. Progeny lines obtained by self-pollination had larger leaves and leaf shapes intermediate of the parental species. Pollen fertility was gradually recovered in the first and second progenies. The backcrossing progeny lines, as a whole, exhibited morphologies were similar to Chinese cabbage. Morphological variations were observed among the somatic hybrids and their progenies.
Brassica napus ; genetics ; growth & development ; Breeding ; Chromosomes, Plant ; Hybridization, Genetic ; genetics ; Mustard Plant ; genetics ; growth & development ; Plant Somatic Embryogenesis Techniques ; Ploidies ; Pollen ; physiology ; Protoplasts ; cytology ; Random Amplified Polymorphic DNA Technique ; Recombination, Genetic

OBJECTIVETo compare the photosynthetic characteristics difference of different ploidy Rhodiola sachalinensis germplasm and provide the scientific basis for their cultivation.

METHODLI-6400/XT photosynthesis system was used to measure leaf light response curve and CO2 response curve of diploid and autotetraploid. Biomass, leaf area, stomatal characteristics and chlorophyll content differences were compared in the study.

RESULTStomata of the two germplasms were open during daytime obviously, and stomata conductance responded to the changes of light intensity and CO2 concentration which was not consistent with the characteristics of CAM (crassulacean acid metabolism) plants. Light compensation point of autotetraploid was significantly lower than that of the diploid, and light saturation points of both germplam were close, and their light saturation points were near 500 micromol x m(-2) x s(-1). Quantum efficiency of autotetraploid was significantly higher than the diploid, and the net photosynthetic rate of autotetraploid significantly higher than the diploid when light intensity was higher than 500 micromol x m(-2) x s(-1). Stomata conductance, transpiration rate of autotetraploid was also significantly higher than that of diploid. Biomass, leaf area, stomata diameter and chlorophyll content of autotetraploid were much higher than that of diploid, while the stomata density of autotetraploid was less than diploid.

CONCLUSIONThe results above provide scientific basis for the cultivation of different ploidy Rh. sachalinensi germplasm.

Carbon Dioxide ; metabolism ; Photosynthesis ; physiology ; Ploidies ; Rhodiola ; metabolism

BACKGROUND: Study on epigenetics of urothelial carcinomas has expanded and allowed better understanding of their correlation with clinicopathologic features. The aim of this study was to determine reliable predictive epigenetic markers for patients with urothelial carcinoma of urinary bladder. METHODS: In 64 urothelial carcinomas of the urinary bladder, methylationspecific polymerase chain reaction with RAS association domain family 1A (RASSF1A), adenomatous polyposis coli (APC), death-associated protein-kinase (DAPK), runt-related transcription factor 3 (RUNX3), p14, p16 and MGMT was performed and correlated the results with p53 mutations, DNA ploidy, clinicopathologic parameters and recurrences. RESULTS: Hypermethyation of RASSF1A, APC, DAPK, RUNX3, p14, p16 and MGMT promoters was observed in 35 (54.7%), 29 (45.3%), 18 (28.1%), 18 (28.1%), 9 (14.1%), 2 (3.1%), and 6 (9.4%) cases, respectively. Hypermethylation of RUNX3 and APC was significantly associated with high histologic grades and aneuploidy. Methylation of DAPK was significantly associated with muscle invasion. Methylation of DAPK and RUNX3 genes was significantly associated with recurrence. In survival analyses, methylation of RUNX3 gene and methylation-high (methylation at two or more loci) phenotype was significantly associated with poor recurrence-free survival. CONCLUSIONS: Methylation of RUNX3 gene and methylation-high phenotype are significant indicator of recurrence.
Adenomatous Polyposis Coli ; Aneuploidy ; DNA ; Epigenomics ; Genes, Tumor Suppressor ; Humans ; Methylation ; Muscles ; Phenotype ; Ploidies ; Polymerase Chain Reaction ; Prognosis ; Recurrence ; Transcription Factor 3 ; Urinary Bladder

Microbial lipopeptides play an important role in apoptosis induction of tumor cells. However, there is little knowledge about the relationship between apoptosis induction and membrane fatty acids. The present study focused on the effects of lipopeptides produced by Bacillus subtilis HSO121 on Bcap-37 cell lines. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl (MTT) colorimetric assay and surface tension measurements, showed that the critical micelle concentration (CMC) was a critical level for the inhibitory activity of lipopeptides on the growth of Bcap-37 cells. Under the CMC, the order of least to greatest cytotoxicity effect on cancer cell lines by lipopeptides is C(13)-lipopeptide < C(14)-lipopepitde < C(15)-lipopeptide. Above CMC, all lipopeptides directly exert cytolytic activity. The flow cytometric analysis and Hoechst33258 staining experiments confirmed the apoptosis of Bcap-37 cell lines induced by lipopeptides in a dose-dependent manner. This apoptosis was associated with a significant decrease of the unsaturated degree of the cellular fatty acids of Bcap-37 cell lines due to the changes in the cellular fatty acids composition induced by the lipopeptide treatment. These results indicated that disturbance of the cellular fatty acid composition of breast cancer cell lines were related to in the cell apoptosis. Furthermore, significant difference in IC(50) values of tumor cells and normal cell showed that the lipopeptide exerted selective cytotoxicity on the cancer cells. Thus HSO121 lipopeptides may have potential applications as an anticancer leads.
Antineoplastic Agents ; chemistry ; pharmacology ; Apoptosis ; Bacillus subtilis ; Cell Cycle ; Cell Membrane ; chemistry ; metabolism ; Cell Survival ; drug effects ; Fatty Acids ; chemistry ; metabolism ; HeLa Cells ; Humans ; Lipopeptides ; chemistry ; pharmacology ; Ploidies ; Tumor Cells, Cultured