Bilineage embryo-like structure from EPS cells can produce live mice with tetraploid trophectoderm

Self-organized blastoids from extended pluripotent stem (EPS) cells possess enormous potential for investigating postimplantation embryo development and related diseases. However, the limited ability of postimplantation development of EPS-blastoids hinders its further application. In this study, single-cell transcriptomic analysis indicated that the "trophectoderm (TE)-like structure" of EPS-blastoids was primarily composed of primitive endoderm (PrE)-related cells instead of TE-related cells. We further identified PrE-like cells in EPS cell culture that contribute to the blastoid formation with TE-like structure. Inhibition of PrE cell differentiation by inhibiting MEK signaling or knockout of Gata6 in EPS cells markedly suppressed EPS-blastoid formation. Furthermore, we demonstrated that blastocyst-like structures reconstituted by combining the EPS-derived bilineage embryo-like structure (BLES) with either tetraploid embryos or tetraploid TE cells could implant normally and develop into live fetuses. In summary, our study reveals that TE improvement is critical for constructing a functional embryo using stem cells in vitro.

The chemical reprogramming of unipotent adult germ cells towards authentic pluripotency and de novo establishment of imprinting

Although somatic cells can be reprogrammed to pluripotent stem cells (PSCs) with pure chemicals, authentic pluripotency of chemically induced pluripotent stem cells (CiPSCs) has never been achieved through tetraploid complementation assay. Spontaneous reprogramming of spermatogonial stem cells (SSCs) was another non-transgenic way to obtain PSCs, but this process lacks mechanistic explanation. Here, we reconstructed the trajectory of mouse SSC reprogramming and developed a five-chemical combination, boosting the reprogramming efficiency by nearly 80- to 100-folds. More importantly, chemical induced germline-derived PSCs (5C-gPSCs), but not gPSCs and chemical induced pluripotent stem cells, had authentic pluripotency, as determined by tetraploid complementation. Mechanistically, SSCs traversed through an inverted pathway of in vivo germ cell development, exhibiting the expression signatures and DNA methylation dynamics from spermatogonia to primordial germ cells and further to epiblasts. Besides, SSC-specific imprinting control regions switched from biallelic methylated states to monoallelic methylated states by imprinting demethylation and then re-methylation on one of the two alleles in 5C-gPSCs, which was apparently distinct with the imprinting reprogramming in vivo as DNA methylation simultaneously occurred on both alleles. Our work sheds light on the unique regulatory network underpinning SSC reprogramming, providing insights to understand generic mechanisms for cell-fate decision and epigenetic-related disorders in regenerative medicine.

Variedades de cultivo de líquido amniótico para el diagnóstico in vitro de poliploidías / Amniotic fluid culture varietys for poliploydies in vitro diagnosis

RESUMEN Fundamento: El cultivo celular permite el análisis directo de las células vivas mediante un microscopio. El estudio de las células contenidas en el líquido amniótico, mediante técnicas de cultivo, detecta anomalías en número y morfología de los cromosomas, que pueden relacionarse con enfermedades genéticas. Objetivo: Caracterizar las variedades de cultivo de líquido amniótico para el diagnóstico in vitro de poliploidías. Metodología: Se realizó un estudio descriptivo transversal, en el Centro Provincial de Genética Médica de Camagüey, en el periodo de noviembre de 2016 a abril de 2018.La población de estudio estuvo constituida por 1571 muestras útiles de líquido amniótico obtenidas por amniocentesis, en gestantes en el segundo trimestre, evaluadas en consulta multidisciplinaria con criterios clínicos de estudios cromosómicos según lo establecido en el diagnóstico prenatal citogenético, previo consentimiento informado. Se utilizaron 20 mL de líquido amniótico para la siembra de células fetales, y se aplicaron tres variantes de cultivo abierto (directo, centrifugado y expandido). Se determinó el complemento cromosómico en cada variedad. Resultados: Predominó el complemento cromosómico normal. Las tetraploidías prevalecieron en el cultivo expandido. El índice mitótico fue similar en las tres variedades de cultivo y el cultivo directo tuvo el más bajo índice de poliploidías. Conclusiones: El cariotipo normal fue predominante. Las tetrapolidías fueron las alteraciones más frecuentes y prevalecieron en el cultivo expandido. En el cultivo directo se presentó el más bajo índice de errores inducidos in vitro.

ABSTRACT Background: Cell culture allows direct analysis of live cells under a microscope. The cell study contained in amniotic fluid, by culture techniques, detects abnormalities in chromosome number and morphology, which can be related to genetic diseases. Objective: To describe amniotic fluid culture strains for the in vitro diagnosis of polyploidy. Methodology: A cross-sectional descriptive study was conducted at the Camagüey Provincial Center of Medical Genetics, from November 2016 to April 2018.The study population consisted of 1571 useful amniotic fluid samples obtained by amniocentesis, in pregnant women in the second trimester, evaluated by multidisciplinary discussion with clinical criteria for chromosomal studies as established in the cytogenetic prenatal diagnosis, prior informed consent. 20 mL of amniotic fluid were used for fetal cell seeding, and three open culture strains (direct, centrifuged and expanded) were applied. Chromosomal complement was determined in each variety. Results: Normal chromosome complement was predominant. Tetraploidy prevailed in the expanded culture. The mitotic index was similar in the three culture strains and the direct culture had the lowest polyploidy index. Conclusions: Normal karyotype was predominant. Tetraploidy were the most frequent modifications and prevailed in the expanded culture. Direct culture had the lowest rate of the in vitro induced errors.

Dry matter productivity and bromatological quality of ryegrass genotypes cultivated in southern Brazil / [Produtividade de matéria seca e qualidade bromatológica de genótipos de azevém cultivados no Sul do Brasil]

The goal of this work was to assess the biomass production and bromatological quality of ryegrass genotypes in ten municipalities of the Western and North Plateau regions of the State of Santa Catarina, Brazil. The cultivars La Estanzuela 284 (diploid), Bar HQ, Barjumbo, INIA Escorpio, Potro, and Winter Star (tetraploids) were compared, distributed in a randomized block design, in which the municipalities constituted the blocks, with three replications. The cuts were performed when the plants reached 30cm, leaving a residue of 10cm. In three cuts, the cultivars Barjumbo and Bar HQ were the most productive, exceeding 4.6 t ha-1 of dry matter. In the places in which five cuts were performed, the production of these cultivars exceeded 7.3 t ha-1, placing them again ahead of the others. The average crude protein content in three cuts was greater than 25% in all cultivars. There was no difference between the genotypes in the content of neutral detergent fiber and total digestible nutrients. There was a significant correlation between quantitative and qualitative productive variables. The assessed cultivars represented good options for composing short-term or long-term winter-feeding systems, adjusted to the integration with annual crops or warm-season pastures.(AU)

O objetivo deste trabalho foi avaliar a produção de biomassa e qualidade bromatológica de genótipos de azevém, em dez municípios das regiões Oeste e Planalto Norte Catarinense, Estado de Santa Catarina, Brasil. Foram comparados os cultivares La Estanzuela 284 (diploide), Bar HQ, Barjumbo, INIA Escorpio, Potro e Winter Star (tetraploides), distribuídos em um delineamento blocos casualizados, em que os municípios constituíram os blocos, com três repetições. Os cortes foram realizados quando as plantas atingiram 30cm, deixando um resíduo de 10cm. Sob três cortes, os cultivares Barjumbo e Bar HQ foram os mais produtivos, ultrapassando 4,6 t ha-1 de matéria seca. Nos locais em que ocorreram cinco cortes, a produção destes cultivares superou 7,3 t ha-1, posicionando-os novamente à frente dos demais. O teor médio de proteína bruta em três cortes foi superior a 25% em todos os cultivares. Não houve diferença entre os genótipos no teor de fibra detergente neutro e de nutrientes digestíveis totais. Verificou-se correlação significativa entre variáveis produtivas quantitativas e qualitativas. Os cultivares testados representam boas opções para compor sistemas forrageiros hiberno-primaveris de curta ou longa duração, ajustando-se à integração com lavouras ou pastagens anuais de estação quente.(AU)

Analysis of genetic and clinical characteristics of nine cases of myelodysplastic syndrome with near tetraploid/tetraploidy karyotype / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic and clinical characteristics of near-tetraploidy/tetraploidy karyotype (NT/T) in patients with myelodysplastic syndrome (MDS).@*METHODS@#Cytogenetic findings of 1576 inpatients with primary MDS were retrospective analyzed, among which 9 were diagnosed with NT/T. Clinical data including gender, age, morphology, genetic feature and prognosis were analyzed.@*RESULTS@#The prevalence of MDS patients with NT/T (NT/T-MDS) among all cases was 0.57%. Karyotyping analysis suggested that eight MDS patients had sole NT/T, while the remainder one had a complex karyotype. In addition to the typical morphology of MDS, NT/T-MDS had unique morphology including huge blast, double-nuclear cell and irregular nuclear membrane. One NT/T-MDS patient gave up therapy, and the remaining eight underwent the first course of treatment, albeit with poor prognosis. Only one patient had complete remission, one had partial remission, three had no remission; and three had converted to acute myeloid leukemia.@*CONCLUSION@#NT/T-MDS is rare and has unique morphology. Generally, NT/T-MDS patients have poor prognosis. However, NT/T cannot be simply classified as high-risk group, but with consideration whether they have affected particular chromosomal structures as well as other clinical data.

Imprinting genes modified parthenogenetic embryonic stem cells produce full-term mouse via tetraploid complementation / 生物工程学报

Parthenogenetic embryonic stem cells (pESCs) derived from bi-maternal genomes do not have competency of tetraploid complementation, due to lacking of paternal imprinting genes. To make pESCs possess fully development potentials and similar pluripotency to zygote-derived ESCs, we knocked out one allelic gene of the two essential maternal imprinting genes (H19 and IG) in their differentially methylated regions (DMR) via CRISPR/Cas9 system and obtained double knock out (DKO) pESCs. Maternal pESCs had similar morphology, expression levels of pluripotent makers and in vitro neural differentiation potentials to zygotes-derived ESCs. Besides that, DKO pESCs could contribute to full-term fetuses through tetraploid complementation, proving that they held fully development potentials. Derivation of DKO pESCs provided a type of major histocompatibility complex (MHC) matched pluripotent stem cells, which would benefit research in regenerative medicine.

Inhibitory effect of the aqueous extract of a tetraploid ‘etteum’ variety in Platycodon grandiflorum on degranulation and inflammatory mediator release in RBL-2H3 cells / 한국영양학회지

PURPOSE: Platycodon grandiflorum (a domestic diploid variety, DV-PG) has been used as a food and component of various traditional oriental medicines. Although DV-PG is known to have an anti-allergic effect, little is known about the beneficial health effects of the tetraploid ‘Etteum’ variety in the Platycodon grandiflorum (TV-PG), which is a recently developed variety. In this study, we investigated the effect of TV-PG on the rat basophilic leukemia mast cell (RBL-2H3)-mediated allergic response. METHODS: To examine the effects of TV-PG on the allergic response, RBL-2H3 cells were sensitized with dinitropheny (DNP)-immunoglobin E, treated with various concentrations of TV-PG, and challenged with DNP-human serum albumin. We estimated cell granulation by measuring the release of β-hexosaminidase and production of inflammatory mediators by ELISA. RESULTS: TV-PG had no effect on the proliferation or cytotoxicity of RBL-2H3 cells within the concentration range of 0 to 200 µg/mL. TV-PG inhibited degranulation of RBL-2H3 cells by antigen stimulation in a dose-dependent manner. TV-PG also suppressed the production of inflammatory cytokines and mediators such as interleukin-4, tumor necrosis factor-α, prostagladin E2, and leukotriene B4 in RBL-2H3 cells by antigen stimulation. CONCLUSION: These results indicate that TV-PG exhibits anti-allergic activity via inhibition of degranulation as well as suppression of inflammatory mediators and cytokine release. These findings suggest that TV-PG may have potential as a preventive and therapeutic agent for the treatment of various allergic diseases.

Cytogenetic and molecular genetic analysis of the amniotic fluid cells of a fetus with pseudodicentric isochromosome 22 resulting in partial tetraploidy of 22q / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To diagnose chromosomal abnormalities in amniotic fluid cells by combining karyotyping and single nucleotide polymorphism array (SNP-array) analysis, and to explore the application of SNP-array in routine clinical practice.</p><p><b>METHODS</b>Conventional G banding was used to karyotype a fetal amniotic fluid sample and the corresponding peripheral blood samples from the parents, followed by SNP-array analysis of the fetal genomic DNA from the amniotic fluid.</p><p><b>RESULTS</b>The karyotype of the amniocytes was 47, XX, +mar. The marker chromosome was further identified as psu idic (22) (q11.2) by SNP-array analysis, revealing tetraploidy of a 1.7 Mb fragment in 22q11.1-q11.2 interval that involves the critical region for Cat eye syndrome.</p><p><b>CONCLUSION</b>A rare chromosomal abnormality was identified by combining conventional G banding and SNP-array. The high resolution SNP-array could provide more detailed information for determining the origin of chromosomal abnormalities.</p>

Características estruturais de cultivares diplóides e tetraplóides de azevém / Structural characteristic of diploid and tetraploid ryegrass cultivars

O objetivo do estudo foi determinar, em casa de vegetação, as características estruturais de cultivares de azevém diplóides e tetraplóides, em regime de corte. Os tratamentos constaram de três cultivares diploides [Comum-RS (Lolium multiflorum), Pronto® (L. multiflorum var. westerwoldicum) e Conquest® (L. multiflorum var. italicum)]; e quatro tetraploides [INIA Titan® (L. multiflorum var. italicum), Winter Star® (L. multiflorum var. westerwoldicum), KLM 138® (L. multiflorum var. italicum) e Banquet II® (L. perenne)], alocados em delineamento completamente ao acaso com seis repetições. As cultivares foram semeadas na densidade de 10 sementes vaso-1 de 2500g de solo, com correção e fertilização realizada em dose única antes da semeadura. Quando as plantas atingiram 20 cm de altura foi realizado o primeiro corte, para dar condições ao adequado estabelecimento, enquanto os demais foram realizados quando era atingida altura média de 15 cm, deixando-se sempre resíduo de 7 cm. Por análise de variância e comparação de médias pelo teste de Tukey (P<0,05), foram analisadas as variáveis estruturais: Comprimento final da folha, número de folhas vivas por perfilho, densidade populacional de perfilhos, comprimento da planta e relação folha:colmo, avaliadas no dia de cada corte, momentos antes da execução deste. As cultivares Comum-RS, INIA Titan®, Winter Star®, Conquest®, KLM 138®, Pronto® e Banquet II® diferem quanto às características estruturais estudadas. A cultivar Banquet II® apresenta as melhores características estruturais, todavia as cultivares Winter Star®, Conquest® e KLM 138® também apresentam estrutura adequada ao pastejo durante todo seu ciclo, enquanto Pronto® e Comum-RS, ao final do ciclo, têm a acessibilidade das folhas comprometida.

The main goal was to determine, under defoliation, in greenhouse, ryegrass structural traits of diploid and tetraploid cultivars. Treatments consisted of three diploid cultivars [Comum-RS (Lolium multiflorum), Pronto® (L. multiflorum var. westerwoldicum) and Conquest® (L. multiflorum var. Italicum)], and four tetraploids [INIA Titan® (L. multiflorum var. Italicum), Winter Star® (L. multiflorum var. westerwoldicum), KLM 138® (L. multiflorum var. italicum) and Banquet II® (L. perenne)], in a completely randomized design with six replications. Cultivars were sown at a density of 10 seeds pot-1 2500g of soil; correction and fertilization was performed in a single dose before sowing. When plants reached 20 cm height the first cutting was made, to allow for appropriate establishment. The following cuttings were made when when plants reached 15 cm average height, always leaving a residue of 7 cm. Data were submitted to analysis of variance and means compared by Tukey test (P<0.05). Variables evaluated were: Final length leaf, number of life leaves, population tillers density, plant length and leaf/sheath ratio, evaluated on each cut, moments before the execution of this. Cultivars Comum-RS, INIA Titan®, Winter Star®, Conquest®, KLM 138®, Pronto® e Banquet II® differ as to structural characteristics studed. Banquet II® cultivar presents the best structural features, however the Winter Star®, Conquest® and KLM 138® cultivars also have adequate structure to grazing throughout your cycle, while Pronto® and Comum-RS, to the end of the cycle, have accessibility leaves compromised.

Morphology and AFLP analysis of tetraploid plantlets of Atractylodes macrocephala / 中国中药杂志

In order to investigate the genetic basis of morphological variation of tetraploid plantlets of Atractylodes macrocephala, diploid plantlets were taken as experimental material, sterile filtration colchicine was used to soak 0.5-1.0 cm long buds. The difference between morphology and stomatal of diploid and tetraploid of A. macrocephala was compared, and genome polymorphism was explored by AFLP. The results showed that the buds dipped in 0.1% colchicine solution for 36 h was optimal conditions to induce tetraploid of A. macrocephala with induction rate of 32.0%. Morphological indexes such as leaf area index, leaf length and width, the density of stomas and the number of chloroplast of tetraploid were distinctly different from diploid. Four hundred and fifty-one bands ranging with 80-500 bp were amplified with 24 pairs of primers, the rate of polymorphism was 32.59%. These amplification sites of diploid were different from tetraploid of A. macrocephala, and the differences in morphology of them were reflected in the DNA polymorphism.

Two Cases of Near-Tetraploidy in Acute Leukemias of Ambiguous Lineage

No abstract available.

Optinization of rapid propagation technique and induction and identification of autotetraploid of Polygonum multiflorum / 中国中药杂志

<p><b>OBJECTIVE</b>To establish and optimize the rapid propagation system of Polygonum multiflorum, as well as explore method for induction and identification of autotetraploid.</p><p><b>METHOD</b>Propagation medium was optimized by orthogonal test. The buds were immersed in colchicine solution with different concentrations for different time to select induction conditions for autotetraploid of P. multiflorum.</p><p><b>RESULT</b>The most appropriate propagation medium was MS medium supplemented with 1.0 mg x L(-1) 6-BA, 0.3 mg x L(-1) NAA, and 0.4 mg x L(-1) PP333. That the buds were soaked in 0.2% colchicine solution for 30 h, or soaked in 0.3% colchicine solution for 18 h, was optimal condition to induce autopolyploid of P. multiflorum with induction rate as high as 16.7%.</p><p><b>CONCLUSION</b>Rapid propagation of P. multiflorum could be achieved by tissue culture. Furthermore, colchicine was an effective inducer of polyploidy, and 25 tetraploid lines were obtained through chromosome identification. The experiment laid a foundation for the wild resource conservation, superior varieties breeding of P. multiflorum.</p>

Effect of gas-turbine green discoloring and drying processing methods on herbal quality of tetraploid Lonicerae Japonicae Flos / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of gas-turbine green discoloring and drying processing method on the quality of various Lonicerae Japonicae Flos herbs.</p><p><b>METHOD</b>DIKMA DiamonsilTM-C18 column (4.6 mm x 250 mm, 5 microm) was adopted using HPLC Waters 1525 and eluted with acetonitrile and 0.1% phosphate acid as the mobile phase. The flow rate was 1.0 mL x min(-1) , the column temperature was 25 degrees C the detection wavelength was 355 nm.</p><p><b>RESULT</b>After being processed by the gas-turbine green discoloring and drying method, tetraploid Lonicerae Japonicae Flos showed a green color. The contents of chlorogenic acid and galuteolin were 5.31% and 0.105% , both significantly higher by 18.0% and 32.1% than those of diploid Lonicerae Japonicae Flos processed by the same method. The content of chlorogenic acid in tetraploid Lonicerae Japonicae Flos processed the gas-turbine green discoloring and drying method were also remarkably higher than that of tetraploid and diploid Lonicerae Japonicae Flos processed by traditional processing method of natural drying.</p><p><b>CONCLUSION</b>The gas-turbine green discoloring and drying processing method is a new-type drying method suitable for tetraploid Lonicerae Japonicae Flos. Under the condition of gas-turbine green discoloring and drying processing, tetraploid Lonicerae Japonicae Flos shows much higher quality than Lonicerae Japonicae Flos, suggesting that it is a good variety worth popularizing and applying.</p>

Chromosomal Abnormalities in Myelodysplastic Syndrome with Near-tetraploidy: A Case Report

Massive hyperdiploidy and tetraploidy are rare cytogenetic abnormalities in myelocytic malignancies, especially in myelodysplastic syndrome (MDS). These abnormalities are known to be associated with leukemogenesis, leukemic transformation and poor prognosis. We report here the first case of MDS with near-tetraploid cytogenetic abnormality in Korea. A 80-yr-old male was diagnosed with refractory anemia with excess blasts-2 (RAEB-2). Bone marrow aspiration smear showed 16% of blasts, which were large sized myeloid blasts with irregular margins and cytoplasmic vacuolation. Cytogenetic analysis of bone marrow cells revealed numercal and structural cytogenetic abnormalities including near-tetraploidy in 8 of 20 metaphases: 45,XY,add(1)(p36.1),del(10)(p11.2),del(11)(q13),-12,-16,der(17)t(11;17) (q13;q21),add(20)(q13.1),+mar[8]/85~90,idemx2[cp8]/46,XY[4]. After chemotherapy with decitabine, he showed pancytopenia during follow-up period and died of sepsis 14 months after the diagnosis.

Blastoid Mantle Cell Lymphoma: A Case Report / 대한피부과학회지

Mantle cell lymphoma (MCL) is a low-grade malignant lymphoma derived from a subset of B-cells localized in the mantle zone of the lymphoid follicle. MCL commonly involves extranodal sites, usually as a manifestation of disseminated disease. The immunophenotype of MCL is characterized as CD20+, CD5+ and Cyclin D1+. Blastoid mantle cell lymphoma (BV-MCL) is a rare variant and has a more aggressive clinical course. In the blastoid variant, the genetic profile is often more complex and includes tetraploid chromosomes, CCND1 amplification, a higher incidence of p53 gene mutations, and p16(INK4a) deletions. We report a case of a 62-year-old male patient who presented with erythematous subcutaneous nodules on the lower back. He had been treated for BV-MCL of the testis which had occurred 1 year ago. The analysis of histopathological and immunohistochemical examinations of the skin lesions also revealed a BV-MCL.

Location and role of protein kinase Cα in parthenogenetic and tetraploid preimplantation embryonic development in mouse / 生理学报

Protein kinase C (PKC) is a critical molecule in cellular signal transduction in mammals. It is involved in many biological processes in embryonic development, including nuclear remodeling, cell cycle adjustment and cellular polarity regulation. The present study aimed to observe the location of PKCα, an important isozyme of PKC, in fertilized, parthenogenetic and tetraploid preimplantation embryos, and compare the expression of PKCα during embryonic compaction in Kunming mice. The location of PKCα was detected by immunochemistry and laser confocal microscopy. Western blot was performed to quantify PKCα expression during embryonic compaction in the three kinds of embryos. In the experiment, fertilized embryos were flushed from oviduct or uterus at 45, 52, 69, 76 and 93 h after injection of human chorionic gonadotrophin (hCG); parthenogenetic embryos were collected by SrCl2 activation of oocytes for 6 h; and tetraploid embryos were produced by electrofusion of 2-cell embryos. Embryos were fixed at different developmental stages for immunofluorescent staining. 8-cell/4-cell embryos and morula were lysed for Western blot. The results showed that PKCα had similar location pattern in different embryos. It was distributed mainly in the nuclear aggregating around chromatin at different developmental stages. However, PKCα expressed strongly in the interphase than in mitotic blastomere. Before embryonic compaction, PKCα was localized at the blastomere boundary. At late blastocyst stage of fertilized embryos, PKCα was localized only in the polar trophoblast, but not in other trophoblast. At late stage of pathenogenetic blastocyst, there was no clear PKCα signal in the polar trophoblast. Tetraploid embryos had larger blastomere than other embryos and compacted after 4-cell stage, but not after 8-cell stage. Meanwhile, there was PKCα signal at the blastomere boundary at 4-cell stage. Our results showed that the expression of PKCα lasted through all the preimplantation stage. Although there were different expression levels among different stages, the expression increased around embryonic compaction. Quantification of expression of PKCα by Western blot demonstrated that the expression increased after compaction, indicating that the compaction was possibly dependent on the relocation of PKCα. Moreover, it was shown that the second relocation of PKCα occurred during the blastocyst formation. PKCα had different expression patterns in the three kinds of preimplantation embryos. However, the effects of PKCα on embryonic development started in early stage. There must be a necessary connection between PKCα relocation and cell adhesion starting at embryonic compaction.

Histological and quantitative study of the effect of Eruca sativa seed oil on the testis of albino rat

Eruca Sativa [E.S] or Gargir seed oil is widely used in folk medicine. This study was conducted to investigate its possible effect on male rat fertility. Histological changes of the testis, level of testosterone hormone and sperm count were determined. The results revealed that administration of low dose of E.S. seed oil caused dilatation of the seminiferous tubules, proliferation of spermatogenic cells and increase of its mitotic activity. Increased number of sperms and epididymis weight, elevated level of testosterone hormone and hyperplasia of interstitial Leydig cells have also been noticed. DNA analysis revealed an increase of the percentage of haploid and decrease of diploid and tetraploid cells. Administration of E.S. seed oil at higher dose showed. decreased diameter of the seminiferous tubules, reduced spermatogenic activity and number of sperms . Also testosterone hormone level decreased and the interstitial cells appeared few. DNA analysis showed a reduction of the percentage of the haploid and increase of the percentage of diploid and tetraploid cells

Chromosomal Abnormality of Spontaneous Abortus in Relation to Transvaginal Ultrasonographic Finding / 대한산부인과학회잡지

OBJECTIVE: To investigate the importance of transvaginal ultrasonographic findings of spontaneous abortion and the relation of transvaginal ultrasonographic finding and the karyotype of abortus. MATERIALS AND METHODS: Transvaginal sonography was undergone to 48 subjects with spontaneous abortion in their early pregnancies who had visited Ewha Womans University Mokdong Hospital, from March 1996 to June 1999. A chromosome analysis of their aborted products was done. RESULTS: The frequency of chromosome abnormalities of abortus was 47.9%. Among the chromosomal abnormalities, trisomy was 31.2%(simple trisomy 27.0%, double trisomy 4.2%), monosomy X 8.4%, poliploidy 8.4%(tripoidy 4.2%, tetraploidy 4.2%). The average maternal age for trisomy was 30.2+/-4.9, and their past number of spontaneous abortions was 1.0+/-0.9, with trisomy 16 most frequent with 23.0%. In the case with trisomy 5, 22, and double trisomy, the average maternal age was increased with 38.0, 36.0+/-2.8, 35.5+/-5.0, respectively. However, no significant relevance could be found. The average maternal age for polyploidy was 30.5+/-3.7, and their past number of spontaneous abortions was 1.0+/-2.0. No significant relevance could be found compared with normal karyotypes. The average maternal age for monosomy X was 27.5+/-1.7, which is lower than the average maternal age for normal karyotypes, but not significantly so. The mean number of past abortions was no different with 1.0+/-0.8. No significant relevance could be found between ultrasonographic findings and chromosomal abnormalities. The measured/expected crown-rump length ratio of the aborted group was smaller than those of the control group. CONCLUSION: Serial ultrasonographic findings in early pregnancies help to diagnose spontaneous abortion, but not efficient to diagnose chromosomal abnormalities in the aborted group.

A Case of Mosaic Tetrapaloidy

Tetraploidy of human chromosome(92, XXYY) has been described very rarely. Liveborn infant with tetraploidy was reported in only 17 cases(complete 7 cases, mosaicism 10 cases) in the world, and no cases have been reported in Korea. The diploid-tetraploid mosaicism could arise during the early mitosis of the zygote. In one blastomeric cell, the chromosomes replicate but the cytoplasrn does not divide. We report a case of live infant with mosaic tetraploidy detected in lymphocyte(30%) and amniotic cell culture(25%), who presented with multiple congenital anomalies. A brief review of the literature is included.

Correlation of Heregulin mRNA and Her-2/neu Protein Expression with Node Metastasis and DNA Ploidy Pattern in Human Invasive Breast Carcinoma

The Her-2/neu protooncogene encodes a transmembrane tyrosine kinase that is structurally homologous to the receptor for epidermal growth factor. Its amplification and overexpression are associated with poor prognosis in breast cancer patients. Neu differentiation factor is a ligand for Her-2/neu protooncogene and was detected in ras-transformed rat fibroblasts. Heregulin (human homologue of neu differentiation factor) is a 44-kilodalton glycoprotein that stimulates tyrosine phosphorylation and induces growth arrest or stimulation and differentiation in human breast cancer cell lines. In this study we examined the expression of heregulin mRNA by nested reverse transcription (RT) PCR with fresh tissue, Her-2/neu protein, ICAM-1 and steroid receptors by immunohistochemistry, and DNA ploidy pattern by flow cytometry with paraffin-embedded tissue in invasive breast carcinoma. We compared the data with nodal status, lymphovascular invasion, steroid receptor status and DNA ploidy pattern. For RT-PCR to heregulin mRNA, 38 cases of fresh breast cancer tissue were obtained. Total 68 cases of invasive breast carcinoma tissue were fixed in formalin, which were used for routine histology, immunohistochemistry and flow cytometry. The results are as follows; 1) Heregulin mRNA was expressed in 86.1% of patients with invasive breast carcinoma and 100% of patients with benign breast lesion using nested RT-PCR analysis. 2) Her-2/neu protein was overexpressed in 50.0% of tumors using immunohistochemistry. The expression of Her-2/neu protein was significantly correlated with high counts of lymph nodes with metastasis (p<0.05), and high nuclear grade (p<0.05). 3) Her-2/neu protein overexpression was significantly correlated with a high DNA index(p<0.05). All of the tumors showing Her-2/neu protein overexpression and no heregulin mRNA expression revealed near tetraploid DNA content. However, both Her-2/neu overexpression and heregulin mRNA expressing tumors revealed near tetraploidy in 38.9% and diploidy in 50.0%. Based on these results, heregulin mRNA expression rate was 86.1% in human invasive breast carcinoma. Her-2/neu protein overexpression is associated with high positive lymph node number and DNA index. Statistically significant reverse correlation with lymph node metastasis is not present.